FOR THE AWARD OF DEGREE OF DOCTOR OF PHILOSOPHY …shodhganga.inflibnet.ac.in/bitstream/10603/92377/3/03_de... · 2018. 7. 3. · “STUDIES ON FISH HELMINTH PARASITES FROM MARATHWADA

“STUDIES ON FISH HELMINTH PARASITES FROM MARATHWADA REGION OF MAHARASHTRA STATE.”

Thesis submitted to

Dr. Babasaheb Ambedkar Marathwada University Aurangabad.

FOR THE AWARD OF DEGREE OF

DOCTOR OF PHILOSOPHY

IN

ZOOLOGY

Submitted by

MR. PREMCHAND RUPCHAND PARDESHI

Under the guidance

DR. CHANDRASHEKHAR J. HIWARE Ph.D., P.G.D.S., F.Z.S.I., F.S.L.Sc.

Professor

Department of Zoology

Dr. Babasaheb Ambedkar Marathwada University, Aurangabad-431004

Feb- 2010

Respectfully

Dedicated To

My Beloved Parents

Mr. Rupchand Khemchand Pardeshi

Mrs. Rambai Rupchand Pardeshi

Dr. Babasaheb Ambedkar Marathwada University, Aurangabad.


DECLARATION

Date:

We hereby declare that, the present work completed in the form of thesis

entitled “Studies on fish helminth parasites from Marathwada region of

Maharashtra state,” is an original work and has not been submitted or published

before in any form for the fulfillment of any other degree, diploma, associateship

or any other similar title to this or any other University.

Research Guide Research Student Prof. Hiware C.J. Pardeshi P.R.

Dr. Babasaheb Ambedkar Marathwada University, Aurangabad - 431004. M.S., INDIA.


Dr. C.J. Hiware Tele: (O) 91-240-2403399/2403400 Professor Ext. 393/394/395

Fax: 91-240-240335/2403113 Email: [email protected] Tele: (R) 91-240-2400098 Mobile-09423472437

Resi: Radhey, Nipatniranjan Nagar, Hanuman Tekadi, Aurangabad-431 004(India).

Ref. No: Zool-2010 Date: -

CERTIFICATE

This is to certify that the thesis entitled “Studies on fish helminth

parasites from Marathwada region of Maharashtra state” which is being

submitted by Mr. Pardeshi Premchand Rupchand for the degree of Doctor of

Philosophy in Zoology, at Dr. Babasaheb Ambedkar Marathwada University,

Aurangabad is a record of his own work. Carried out under my guidance and

supervision. The matter embodies in this thesis has not been submitted for award

of any other degree, diploma or associateship to this or any other University.

Place: Aurangabad Prof. Hiware C.J.

Date: Ph.D., P.G.D.S., F.Z.S.I., F.S.L.Sc.

ACKNOWLEDGEMENT

I express my sincere gratitude cordial thanks whole hearted respect to my

guide and supervisior Dr. Hiware Chandrashekhar Jalba Professor, Department

of Zoology, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad for

his noble guidance, inspiration, valuable suggestion, keen interest and constant

encouragement throughout the period my research work.

I am immensely thankful to Late Head of Department Dr. B.V. Jadhav

Professor and Head of Department, Dr. Babasaheb Ambedkar Marathwada

University Aurangabad for providing necessary laboratory facilities.

I am greatly to Dr. S.P. Zambare, Professor and Head of Department, Dr.

Babasaheb Ambedkar Marathwada University Aurangabad for providing

necessary laboratory facilities in my research period.

I express heartfelt thanks to all the member of teaching and non teaching

staff for their valuable help and co-operation during my research period.

I express heartly thankful to Principal, Dr. B.C. Ghoble, Vice-Principal,

Dr. U.L.Jagtap., Dr. K.D. Devraroo., Dr. S.B. Dongare., Dr. C.R. Dode., Dr.

Shivaji Ubhahrande., Mr. Balaji Shinde for their inspiration during course of

research.

I am whole heartedly indebted to my beloved parents Mr. Rupchand

Khemchand and Mrs. Rambai Rupchand as well as my brother Ranjit Rupchand

Pardeshi and Mrs. Shobha Ranjit Pardeshi for financial help and co-operation

during course of my research.

I have special thanks to My collegues and friends Dr. Naphade., Dr. R.T.

Pawar, Dr. Salve, Dr. Munde, Mr. Vilas wahule, Mr. Madle, Ganesh Phad,

Kishor Shinde., Sunil Avhad, Sujeet Jamdar, Dr. Yogesh Reddy, Dr. Vasant

Dongare, Dr. Manoranjana Nirmale, Sunil Shinde, Ravindra Bhandare, Atul

Chourpagar, More P.R., Jagtap J.S., Dr. Prashant Joshi, Madhav Waghmare,

Sachin Chauhan, Shantanu Chauhan, Madhav Shinde, Keshav Shinde,

Dnyaneshwar Kute, Mote Pradeep, Sachin Ghoble, Shubhangi Divekar, Subhash

Shingare, Dr. Bhure, Sachin Waghmare, Varsha Dabhade, Bhosle popat, Bhosle

Youraj, Dr. Deepak Gaikwad, Dr. Shinde, Bhandari Jyoty, Gunwanti Arak,

Gajanan Deshmukh, More Babasaheb, Sushil, Tanaji, Pappu Kudnar, Anata

Harkal, Gajanan sontakke, Laxman Gadekar, Amol katkar, Amol Kategaokar,

I must acknowledgement deep appreciation to Mr. B.S. Dhokne photo

artist for the excellent work of photography for my Ph.D. thesis with great

patience skill in time.

Pardeshi Premchand Rupchand

CONTENT

Introduction

Abstract of the Ph.D. Thesis

List of plates and figures

Maps

Photo plates: Host

CHAPTER-I

TAXONOMY

1) Senga rupchandensis n.sp.

2) Senga rambaei n.sp.

3) Circumoncobothrium jadhavae n.sp.

4) Azygia stunkardi Rai, 1964 (Redescribed)

5) Genarchopsis paithanensis n.sp.

6) Phyllodistomum aurangabadensis n.sp

7) Allocreadium khami n.sp.

8) Orientocreadium striatusae n.sp.

CHAPTER-II

HISTOCHEMISTRY


2) Circumoncobothrium jadhavae n.sp




CHAPTER-III HISTOPATHOLOGY






CHAPTER-IV

DNA FINGERPRINTING






SYSTEMATIC POSITION OF THE PARASITES WITH HOST

SYSTEMATIC POSITION OF THE HOST

REFERENCES

INTRODUCTION

The traditions of research in the parasites and parasitism are very old in

this country, but these are relevant even today. There are innumerable exciting

topics on which valuable research contributions could be made. Even the

classical parasitology which includes the study of taxonomy, morphology and

life cycle though much out of fashion these days is no less important and exciting

than the so called modern areas of research.

A parasite is physiologically dependant on its host and cannot survive in

its absence. Parasitism occurs throughout the animal kingdom but the parasitic

species are mostly found among the invertebrates animal such as, protozoans.

Platyhelminthes, nematodes and arthropods. Some zoologist regards parasitism

to be quote close to symbiosis. In both phenomena, the relationship is an intimate

one, in symbiosis both may benefit but not in parasitism. To simply say that the

parasite is an organism that feeds and lives on one in another organisms, obtain

food but the host usually suffer due to effects of their feeding as also due to the

fact that they release metabolic wastes into its body.

Parasitic diseases are among public health problem of tropical countries

including India. They infect man, domestic and wild life. Helminthic infections

are common in the country and endemic in areas with poor socio-economic

status, unhygienic living and food habits morbidity and complication. Many

species of parasites can survive as adult in warm blooded animals and in their

larval stages either in cold blooded animals or may be in water or soil.

Fish culture is a useful and paying proposition, which should help to meet

the present food shortage in the world. As the world becomes more and more

crowded with people, all the food stuffs particularly fish because of its high food

value will become more valuable. Fisheries constitutes the basic food industry as

it is a perfect balance of important nutritional factors such as, vitamins, mineral

and mineral salts etc.

There are different type of infections found in fishes and are caused by

bacteria, protozoan, fungi, helminthic infection etc. The parasites utilize host

energy resources these deleterious effects are frequently nutritional but it is rare

for this to be the only consequence of infection. Any deleterious effects parasites

have as they invade, move around or grow inside or on the host may through

associated pathology, physiology imbalance or general malaise have

consequence that effect the growth, survives and ultimately reproductive finess

of the host. Although parasite infections are likely to affect the behavior of fish

in all aquatic environments. Fish parasites cause commercial losses in both

aquaculture and fisheries industries and may have human as wall as socio-

economic.

Among the recent workers who are working on biochemistry,

histochemistry, histopathology are Bhalerao, Fotedar, Hanumantrao, Gupta,

Cooper, Nama, Pandey, Shinde, Jadhav, Hiware and many other are still working

on different aspects.

In present study, the cestode and trematode parasite collected from the

different region of Marathwada, Maharashtra state, India. The collected worm

were preserved in 4% formalin, washed in distilled water, stained with Harri’s

Haematoxylin, dehydrated in alcoholic grades, cleared in xylol, mounted in DPX.

All drawing were made with Camera Lucida and all measurements taken in

millimeters.

For histochemical studies, the analysis of alkaline and acid phosphatase

activity in the cestode and trematode parasites are carried out by using Gomori’s

method.

In DNA fingerprinting, study of DNA banding pattern from piscian

parasites. The histopathological studies of intestine, liver, buccopharyngeal area

infected by cestode and trematode parasites is carried out.

ABSTRACT OF THE Ph.D. THESIS

Fishes are the apex of predatory-prey pyramid within freshwater as well as

in sea water and therefore tend to be infested by a considerable range of parasites

which occur in large numbers. The economic importance of fish parasites is

related directly to the economic importance of fish they affect. Now it has been

fully realized that fish constitutes an important items of human diet. Fish is an

excellent food which is nutritionally equivalent to meat in protein, low in

saturated fats and high in mineral. The fish production is an important source of

income, employment generation and plays an important role in economy (Akhtar,

1986). During the first part of the 17th century, natural historians and physician

still thought that the few known endoparasite were formed from the excretions

and the bodies of man and other animals. Goeze (1782) and Joerdens (1901) believed that endoparasite helminthes

were beneficial because they consumed the host’s excess foods and intestinal

mucus, which otherwise would putrefy and brings disease. Helminth is an

important group of animal parasites occurring in the adult stage usually in

vertebrate’s host. These worms are widespread in almost all animals in every part

of the worlds, although the intensity of infection may differ from time to time or

place to place and produce a wide variety of direct effects. Thus they play a vital

role on determining the welfare of man and the animal with which it is associated

to smaller and greater extent. The parasites have detrimental effects upon fish in

more ways than one (Srivastav, 1975). Cross (1933) showed that the parasitic

infection tend to decrease the growth rate resulting in the stunting of fish.

Parasite cause damage to various organs of their hosts affecting the yield of fish

products such as, body oil, liver oils etc.

Fish parasites are found in the classes Trematoda, Cestoda, Nematoda and

Acanthocephalan. Aristotle (384-322 B.C.), who wrote “Historia Animalium”,

had stated, earlier. “……… there are three kinds of helminthes: those which one

calls large and flat (Tapeworm), those which are cylindrical (Ascaris

lumbricoides) and the third ones, the Ascaroids (Enterobium vermicularis).

Parasites may be found in all tissues of host, but they are particularly

common on the skin and gills because these external surfaces are easily invaded.

The helminth cause many health hazards and disease among human and animal

population. This leads to major health problems and high economic loss, e.g.

Taeniasis is caused by cestodes, Taenia solium and Taenia saginata.

Man made pollutants and intensification of fish culture resulted in all

increase of environmental changes, which may be stressful to fish (Lio-po and

Lim, 2002). This condition can result in decreased resistance by the fish, causing

spread of disease and parasitic infection (Rottman et al., 1992). Among fish

parasites, the helminth constitutes the major threat to the fish health. Metazoans

parasitic diseases are most common in fishes inhabiting in Indian waters and

encounter more frequently than microbial infection in natural as well as culture

system (Madhavi, 2003). Sometimes mass fish mortality occurs specially in

nursery as well as culture pond and rivers. High stocking density, poor

husbandry, and abundance occurrence of vectors, high organic load and

unfavorable environmental temperature are also equally important contributory

factors for parasitic disease which induce various pathological changes in fishes

(Robert, 2001).

Recent reports suggest that fishes act as a source of serious human

parasitic infectious disease (Dubois & Pearson, 1963; Schnurrenberger, 1975).

Parasites are important in that they affect the productivity of the fish in the

system through metabolites, by decreasing growth rate, reducing the quality of

meat, loss of protein source and making the hosts more susceptible to more or

other pathogenic parasites, i.e. overall loss of economy.

The person gets infected when they eat raw or poorly cooked infected

fish. Infection rates are highest in countries where raw flesh is eaten and

communities that dispose off sewage directly into lake or rivers without proper

treatment, which provide an opportunity for fishes to pick up infection (Hafeez,

2001). Early symptoms in human infection consist of right upper quadrant

abdominal pain, fever, hepatomegaly, biliary colic and with cough, vomiting,

marked Jaundice, generalized abdominal rigidity, diarrhea. The parasitic

infection from man to animal or from animal to man is common.

The prevention of fish getting infected with tapeworm in edemic areas

depends upon controlling the source of infection, proper disposal of sewage and

marketing of fish. Disposal of untreated sewage into water should be prohibited.

Freezing of fish at -10 0C for 24 hours or cooking atleast 10 minutes at 50 0C and

proper drying and pickling of fish kills the larvae. The public should be educated

about the danger of eating raw or improperly cooked fish (Hafeez, 2001). All the

helminth parasite indicates that, these and other parasites are important not only

in producing diseases in fishes but are also important to other group of animals

including human being; which serve as the definitive host for a variety of

parasites. In the present thesis it is decided to work on helminth parasites from

the piscian hosts from Marathwada region of Maharashtra state.

The thesis entitled “Studies on fish helminth parasites from

Marathwada region of Maharashtra state”. The thesis comprises;

1) Taxonomical studies of piscian helminth parasites.

2) Histochemical studies of piscian helminth parasites.

3) Histopathological studies of piscian helminth parasites.

4) Electrophoretic studies of piscian helminth parasites with relevant

bibliography.

The first chapter deals with the taxonomical studies of the helminth

parasites collected from two fishes, namely Mastacembelus armatus (Lecepede,

1800) and Channa striatus (Bloch, 1793). The parasite recovered from these

hosts are belonging to class cestoda and trematoda. The parasite belongs to

Eucestoda, order-Pseudophyllidea, family- Ptychobothridae, Genus-

Circumoncobothrium, one Circumoncobothrium jadhavae n.sp. collected from

the host, Mastacembelus armatus and Genus Senga, two new species; one is

Senga rambaei n.sp. collected from Mastacembelus armatus and one is Senga

rupchandensis n.sp. from Channa striatus. From Digenea, family-Allocreadiidae,

Genus-Allocreadium, one Allocreadium khami n.sp. collected from

Mastacembelus armatus; Genus- Orientocreadium, one Orientocreadium

striatusae n.sp. collected from Channa striatus; family-Gorgoderidae, Genus-

Phyllodistomum, one Phyllodistomum aurangabadensis n.sp. collected from

Channa striatus; family- Hemiuridae, Genus-Genarchopsis, one Genarchopsis

paithanensis n.sp. from Mastacembelus armatus; family-Azygiidae, Genus-

Azygia, one redescribed species, Azygia stunkardi Rai, 1964 from the host

Channa striatus.

The second chapter deals with the histochemical detection of enzymes,

alkaline and acid phosphatase from the Circumoncobothrium jadhavae n.sp.,

Genarchopsis paithanensis n.sp., Allocreadium khami n.sp. collected from

Mastacembelus armatus (Lecepede, 1800) and Senga rupchandensis n.sp.,

Orientocreadium striatusae n.sp. recovered from Channa striatus (Bloch, 1793)

respectively. In the present study, result indicates that, the histochemical

observation of longitudinal section of mature and gravid proglottids of cestode,

Circumoncobothrium jadhavae n.sp. and Senga rupchandensis n.sp. for the

alkaline and acid phosphatase enzyme activity is high in the reproductive organs

and vitellaria, in Genarchopsis paithanensis n.sp., Allocreadium khami n.sp.

Orientocreadium striatusae n.sp., the enzyme activity is also high in

reproductive organ and vitellaria but less in sucker and musculature. Detail of all

is described with microphotographs.

The third chapter deals with the histopathological studies of the tissues of

the host namely, Mastacembelus armatus (Lecepede, 1800) infected with

Circumoncobothrium jadhavae n.sp., Genarchopsis paithanensis n.sp. and

Allocreadium khami n.sp. and Channa striatus (Bloch, 1793) infected with Senga

rupchandensis n.sp. and Orientocreadium striatusae n.sp. The result indicates

that, the transverse section of the liver infected with the Circumoncobothrium

jadhavae n.sp. has cyst and cause enlargement and rupture the hepatocytes of the

liver; Allocreadium khami n.sp. attach to the liver of the host forming cyst;

Genarchopsis paithanensis n.sp. destroy the epithelial layers and approaches to

the villi. Senga rupchandensis n.sp. penetrates deep into the intestinal villi and

damage the mucosa and sub mucosa layer of the host; Orientocreadium

striatusae n.sp. attached and damage the buccopharyngeal area of the host.

The fourth chapter deals with the DNA fingerprinting of parasites;

Circumoncobothrium jadhavae n.sp., Allocreadium khami n.sp., Genarchopsis

paithanensis n.sp. from the host Mastacembelus armatus and Senga

rupchandensis n.sp., Orientocreadium striatusae n.sp. from the host Channa

striatus. From the parasite sample DNA were isolated, purified, and amplified

with the help of PCR and the DNA fragments were seen with the help of

electrophoresis. The result indicates that, the amplification of genomic DNA of

five piscian parasite sample using ISSR marker set were ordered from University

of British Columbia (USB). The present study gave amplification four primer,

811, 812, 814 and 816 for identification. In ISSR analysis 50 fragments ranged

from 05-15 and varied in size from 170 bp to 2230 bp. Primer no. 811, 816

showed the lowest fragments size while the primer no. 814 produced the highest

fragments size. After banding pattern, shows Dendogram analysis and clustering

is discussed in detail.

Research guide Research student

(Prof. Hiware C.J.) (Mr. Pardeshi P.R.)

LIST OF FIGURES AND PLATES

PLATE- 1): HOST

TAXONOMY:

Fig. 1) Senga rupchandensis n.sp.

PLATE: 2) Senga rupchandensis n.sp

Fig. 2) Senga rambaei n.sp.

PLATE: 3) Senga rambaei n.sp.

Fig.3) Circumoncobothrium jadhavae n.sp.

PLATE: 4) Circumoncobothrium jadhavae n.sp.

Fig. 4) Azygia stunkardi Rai, 1964 (R.D.)

PLATE: 5) Azygia stunkardi Rai, 1964 (R.D.)

Fig. 5) Genarchopsis paithanensis n.sp.

PLATE: 6) Genarchopsis paithanensis n.sp.

Fig. 6) Phyllodistomum aurangabadensis n.sp.

PLATE:7) Phyllodistomum aurangabadensis n.sp.

Fig. 7) Allocreadium khami n.sp.

PLATE: 8) Allocreadium khami n.sp.

Fig. 8) Orientocreadium striatusae n.sp.

PLATE: 9) Orientocreadium striatusae n.sp.

PLATE: 10) Alkaline phosphatase enzyme activity in Senga

rupchandensis n.sp.

A) L.S. of mature proglottids showing distribution of alkaline .

. phosphatase enzyme activity.

B) L.S. of gravid proglottids showing distribution of alkaline

phosphatase enzyme activity.

PLATE: 11) Alkaline phosphatase enzyme activity in

Circumoncobothrium jadhavae n.sp.

A) L.S. of mature proglottids showing distribution of alkaline .

. phosphatase enzyme activity

B) L.S. of gravid proglottids showing distribution of alkaline


PLATE: 12) Alkaline phosphatase enzyme activity in Genarchopsis

paithanensis n.sp.

A) L.S. of whole parasite showing distribution of alkaline


B) L.S. of anterior region showing distribution of alkaline


C) L.S. of middle portion showing distribution of alkaline

phosphatase enzyme activity. .

D) L.S. of posterior region showing distribution of alkaline


PLATE: 13) Alkaline phosphatase enzyme activity in Allocreadium

khami n.sp.

A) L.S. of anterior region showing distribution of alkaline


B) L.S. of middle region showing distribution of alkaline


C) L.S. of posterior region showing distribution of alkaline


PLATE: 14) Alkaline phosphatase enzyme activity in Orientocreadium

striatusae n.sp.

A) L.S. of whole parasite showing distribution of alkaline


B) L.S. of anterior region showing distribution of alkaline


. C) L.S. of posterior region showing distribution of alkaline


PLATE: 15) Acid phosphatase enzyme activity in Senga rupchandensis

n.sp.

A) L.S. of mature proglottids showing distribution of acid


B) L.S. of mature proglottids showing distribution of acid

phosphatase enzyme activity (Magnified).

C) L.S. of gravid proglottids showing distribution of acid


PLATE: 16) Acid phosphatase enzyme activity in Circumoncobothrium

jadhavae n.sp.

A) L.S. of mature proglottids showing distribution of acid


B) L.S. of gravid proglottids showing distribution of acid


PLATE: 17) Acid phosphatase enzyme activity in Genarchopsis

paithanensis n.sp.

A) L.S. of whole parasite showing distribution of acid


B) L.S. of anterior region showing distribution of acid


C) L.S. of posterior region showing distribution of acid


PLATE: 18) Acid phosphatase enzyme activity in Allocreadium khami

n.sp.

A) L.S. of anterior region showing distribution of acid


B) L.S. of middle portion showing distribution of acid




PLATE: 19) Acid phosphatase enzyme activity in Orientocreadium

striatusae n.sp.

A) L.S. of whole parasite showing distribution of acid


B) L.S. of anterior region showing distribution of acid




PLATE: 20) Histopathological study of intestine of Channa striatus

(Bloch, 1793).

Fig. 1) T.S. of healthy intestine showing histological structure.

Fig.2) T.S. of intestine showing the deep penetration of scolex of Senga

rupchandensis n.sp.

PLATE: 21) Histopathological study of liver of Mastacembelus

armatus (Lecepede, 1800).

Fig. 1) T.S. of liver showing histological structure.

Fig. 2) T.S. of infected liver showing parasite, Circumoncobothrium jadhavae

n.sp. forming cyst.

PLATE: 22) Histopathological study of intestine of Mastacembelus


Fig. 1) T.S. of healthy intestine showing histological structure.

Fig. 2) T.S. of intestine showing the parasite, Genarchopsis paithanensis n.sp.

approaching the intestinal villi.

Fig. 3) T.S. of intestine showing the parasite, Genarchopsis paithanensis n.sp.

attached to the intestinal villi.

PLATE: 23) Histopathological study of liver of Mastacembelus


Fig. 1) T.S. of liver showing histological structure.

Fig. 2) T.S. of infected liver showing Allocreadium khami n.sp. forming cyst PLATE: 24) Histopathological study of buccopharyngeal

area of Channa striatus (Bloch, 1793).

Fig. 1) T.S. of buccopharyngeal area showing histological structure.

Fig. 2) T.S. of infected of buccopharyngeal area showing Orientocreadium

striatusae n.sp. forming cyst between pharynx and gills.

Fig. 3) T.S. of infected of buccopharyngeal area showing Orientocreadium

striatusae n.sp. forming cyst between pharynx and gills (Magnified).

DNA FINGERPRINTING:






Map of INDIA Showing Maharashtra State Map of Maharashtra showing -- Marathwada Region

Aurangabad

Map of Marathwada showing collection site

Jalna

Latur

Collect ion site: AurangabadJalnaLatur

CHAPTER-I

TAXONOMY

Eucestoda Wardle, McLeod & Radinoky, 1974

Pseudophyllidea Carus, 1863

Ptychobothriidae Luhe, 1902

Senga Dollfus, 1934

Senga rupchandensis n.sp.

INTRODUCTION:

The genus Senga was established by Dollfus, 1934 with its type species S.

bensardi from Betta splendeus, the Siamese fighting fish in an aquarium at

Vincennes, France. S. ophiocephalina Teseng, 1933 as Anchistrocephalus

ophicephalina from Ophicephalus argus at Taimen, China and identified with a

form previously recorded by Southwell, 1913 as Anchistrocephalus polytera

(Anchitrocephalus) Montilli, 1890- syn. Anchistrocephalus Luhe, 1899 from

Ophiocephalus striatus in Bengal, India. S .pycnomera, Woodland, (1934) as

Bothriocephalus pcynomera from Ophiocephalus marulius at Allahabad, India.

Senga lucknowensis Johri, (1956) from Mastacembelus armatus in India.

Fernando and Furtado, (1964) recorded Senga malayana from Channa striatus.

S. parva and S. filiformis from Channa micropeltes at Malacca.

Ramadevi and Hanumantha Rao, (1966) reported the plerocercoid of

Senga species from Panchax panchax. Tadros, (1968) synomised the Genus

Senga with the genus Polynchobothrium and proposed and proposed new

combinations for the species. Furtado and Chaulan, (1971) reported S.

pahangensis from Channa micropeltes at Tesak Bera. Shinde, (1972) redescribed

Senga bensardi from Ophiocephalus gachua in India.

Later Ramadevi and Rao, (1973) described another species S.

vishakapatnamensis in India. Ramadevi, (1976) described the life cycle of S.

vishakhapatnamensis from Ophiocephalus punctatus in a lake at Kondakaria,

A.P., India. Wardle, McLeod and Radinoky, (1974) Senga as distinct genus in the

family Ptychobothidae, Deshmukh, (1980) described new species S. khami from

fresh water fish Ophiocephalus marulius from Kham river at Aurangabad, India.

Jadhav and Shinde, (1980) as described a new species Senga godavari from

Mastacembelus armatus, at Nanded, India. Jadhav and Shinde, 1980 was added

the species S. aurangabadensis from Mastacembelus armatus at Aurangabad,

M.S., India. Kadam et al., (1981) described a new species Senga paithanensis

from intestine of M. armatus. Majid et al., (1984) was added S. raoii and S.

jagannathe from host Channa punctatus. New two species described by Jadhav et

al., 1991 as S. maharashtrii and Senga gachuae from the intestine of

Mastacembelus armatus. Later Monzer Hasnain, 1992 was added S. chauhani

from the host Channa punctatus. Tat and Jadhav, (1997) added a new species

Senga mohekare from the host Mastacembelus armatus at Parli, Dist. Beed,

(M.S.), India.

One more new species is added by Hiware, (1999) as S. armatusae from

Mastacembelus armatus at Pune, M.S., India. Patil and Jadhav, (2003) added new

species S. tappi from M. armatus at Shirpur, Dist. Dhule. Jadhav, (2005) described

the review article of the genus Senga from freshwater fish in India. Pande et al.,

2006 described two new species S. ayodhensis from Amphinuous cuchia and S.

baught from Rita- rita. Bhure et al., (2007) described new species S. jadhavae

from Mastacembelus armatus. Nilima M. Kankale, 2008 described the new

species S. nathsagarensis from the freshwater fish Mastacembelus armatus.

The present communication deals with the description of new species

Senga rupchandensis n.sp. from Channa striatus (Bloch, 1793) and Senga

rambaei n.sp. from Mastacembelus armatus (Lecepede, 1800).

DESCRIPTION:

Five worms were collected from the intestine of Channa striatus (Bloch,

1793) from Godavari River, Shahagad, Jalna district, M.S., India, in the month of

May 2007. cestode were collected or removed from the intestine, washed in

distilled water, flattened between coverglass and slides, fixed in 4% formalin until

24 hours, washed in distilled water, stained with Harri’s Haematoxylene,

dehydrated in ascending series of alcoholic grades (30%, 50% 70% 90% 100%),

cleared in xylene, mounted in DPX. Drawings were made with the help of

Camera Lucida and all the measurement are taken in millimeter.

The worms are long, creamish in colour. The scolex flat, tubular,

cylindrical in shape and measures 0.7159 mm. in length and 0.2386 mm. in

breadth. The scolex bears two bothria overlapping one another, bothria are flat or

elongated sac like structure, it measures (right bothria) 0.4886 mm. in length and

0.1931 mm. in breadth and left bothria measures 0.4545 mm. in length and 0.1477

mm. in breadth. Right bothria is larger than left bothria. The rostellum is flat

having two rows of semicircular hooks, 42-55 in number. Neck is absent.

Mature proglottids are longer than broad; it measures 1.2523 mm. in length

and 0.4514 mm. breadth. Testes are rounded; 350-370 in number and it measures

0.09223 mm. in diameter. Cirrus pouch sac like, oval in shape and it measures

0.05339 mm. in length and 0.03883 mm. in breadth. Cirrus is elongated and

located anterior to genital pore. Genital pore rounded in shape and measures

0.06796 mm. in diameter.

Vagina is elongated, tubular structure and connects with ootypes. Vagina

measures 1.0873 mm. in length and 0.08737 mm. in breadth. The cirrus pouch

overlaps to the uterus. Ootype is circular or rounded between both ovarian lobes.

Ovary bilobed and separated from the Ootype, right ovary lobe measures 0.2184

mm. in length and 0.07766 mm. in breadth, left ovary lobe measures 0.1601 mm.

in length and 0.1213 mm. in breadth. Ootype is rounded and it measures 0.08737

mm. in diameter. Isthmus is located posterior end of the ootype or between the

ovarian lobes; it measures 0.2766 mm. in diameter. Vitellaria are follicular.

Gravid proglottids are broader than mature proglottids. Eggs are oval, non-

operculated and it measures 0.01925 mm. in length and 0.01069 mm. in breadth.

DISCUSSION:

The genus Senga was established by Dollfus, 1934, with its type species

S. bensardi from Betta splendens at Vincennes, France.

Later on described the twenty five species of Senga given are as follows,

1) S. ophicephalina Teseng, 1933 from Ophicephalus argus in Tsinan,

China.

2) S. bensardi Dollfus, 1934 from Betta splendeus in France.

3) S. pcynomera Woodland, 1934 from Ophiocephalus marulius in India.

4) S. lucknowensis, Johri, 1956 from Mastacembelus armatus in India.

5) S. malayana Furnando and Furntado, 1964 from Channa striatus in

Malacca.

6) S. parva Furnando and Furtado, 1964 from Channa micropeltes in

Malacca.

7) S. pahanensis Furtado et al., 1971 from Channa micropeltis in Tasek,

Bera.

8) S. vishakhapatnamensis Ramadevi et al., 1973 from Ophiocephalus

punctatus in India.

9) S. khami Deshmukh and Shinde, 1980 from Ophiocephalus marulius in

India.

10) S. aurangabadensis Jadhav et al., 1980 from Mastacembelus armatusin

India.

11) S. godavari Shinde et al., 1980 from Mastacembelus armatus in India.

12) S. paithanesis Kadam et al., 1981 from Mastacembelus armatus in

Paithan, India.

13) S. raoii Majid M.A. and G.B. Shinde, 1984 from Channa punctatus in

India.

14) S. jagannathae M.A. Majid and G.B. Shinde, 1984 from Channa

punctatus in India.

15) S. gachuae Jadhav et al., 1991 from Mastacembelus armatus in Solapur

(M.S.), India.

16) S. maharashtrii Jadhav and Tat, 1991 from Mastacembelus armatus in

Amravati, India.

17) S. chauhani Monzer Hashain, 1992 from Channa punctatus in

Janshedpur (M.S.), India.

18) S. mohekarae Tat and Jadhav, 1997 from Mastacembelus armatus in

Osmanabad, India.

19) S. armatusae C.J. Hiware, 1999 from Mastacembelus armatus in Pune.

20) S. tappi D.N. Patil & B.V. Jadhav 2003 from Mastacembelus armatus in

Shirpur, India

21) S. ayodhenensis Pande et al., 2006 from Amphinuous cuchia in India.

22) S. baught Pande et al., 2006 from Rita- rita in India.

23) S. jadhavae Bhure et al., 2007 from Mastacembelus armatus in India.

24) S. nathsagarensis Nilima M. Kankale, 2008 from Mastacembelus

armatus in India

The present worm under discussion is scolex tubular, cylindrical, bears

two bothria, hooks semicircular having 42-55 in number, neck absent, cirrus

pouch sac like, oval, curved, testes oval or rounded having 350-370 in number,

uterus elongated and overlaps to the cirrus pouch, Ootype circular, rounded,

ovary bilobed, genital pore large, vitellaria follicular, eggs oval and non

operculated.

The present worms differ from the species S. ophicephalina Teseng, 1933

from Ophiocephalus argua in China which is having scolex is (cylindrical vs

pear shaped), testes (350-370 vs 50-55) in number, vitellaria lobulate.

S. bensardi Dollfus, 1934 from Betta splendens in France which is scolex

(tubular or cylinder vs triangular), hooks 50 in number, testes (350-370 vs 160-

175) in number, vitellaria (follicular vs granular).

The present worm differs from the species S. pcynomera Woodland, 1934

from Ophiocephalus marulius in India which is having scolex is (tubular or

cylindrical vs elongated), hooks (42-55 vs 68) in number, mature proglottides are

indistinct, ovary discontinuous in two groups, testes (350-370 vs 120-150) in

number, vitellaria (follicular vs granular).

The present worm differs from the species S. lucknowensis Johri, 1956

from Mastacembelus armatus in India which is having scolex (tubular or

cylindrical vs pear shaped), hooks (42-55 vs 36-48) in number, testes (350-370

vs 100-150) in number, vitellaria lobulate and discontinuous two groups.

The present parasites differ from the species S. malayana Furnando and

Furtado, 1964 from Channa striatus in Malacca. Which is having scolex (tubular,

cylindrical vs circular), hooks (42-55 vs 60) in number, testes (350-370 vs 120-

150) in number, vitellaria (follicular vs lobate).

The present parasites differ from the species S. parva Furnando and

Furtado, 1964 from Channa micropeltis in Malacca in the presence of scolex

(tubular, cylindrical vs pear shaped), hooks (42-55 vs 38-40) in number, testes

(350-370 vs 150-180) in numbers, vitellaria are (follicular vs granular).

The present parasite differs from the species S. pahanensis Furtado et al.,

1971 from Channa micropeltis in Tasek, Bera which is having scolex s (tubular,

cylindrical vs triangular), neck is (absent vs present), testes testicular (not lobed

vs lobed) and vitellaria (follicular vs lobulated).

The present worms differ from the species S. visakhapatanamensis

Ramadevi et al., 1973 from Channa punctatus in India, in having scolex (tubular,

cylindrical vs circular), hooks (42-55 vs 50-55), and testes (350-370 vs 40-55) in

number, vitellaria (follicular vs lobulated).

The present parasites differ from the species S. khami Deshmukh and

Shinde, 1980 from Ophiocephalus marulius in India which is having scolex

(tubular, cylindrical vs rectangular), hooks (42-55 vs 55-57) in number, neck

present, testes (350-370 vs 155) in number..

The present parasite or worm differs from the species S. aurangabadensis

Jadhav et al., 1980 from Mastacembelus armatus in India which is having scolex

is (tubular, cylindrical vs oval), hooks 50-52 in number, testes (350-370 vs 240-

260) in number.

The present worm differs from the species S. godavarii Shinde et al., 1980

from Mastacembelus armatus in having scolex (tubular, cylindrical vs pear

shaped), hooks 42-55 in number, testes (350-370 vs 230) in number, vitellaria

follicular with 3-4 rows.

The present worm differs from the species S. paithanensis Kadam et al.,

1981 from Mastacembelus armatus in India. Which is having scolex (tubular,

cylindrical vs triangular), hooks 54 in number, neck (absent vs present), testes

(350-370 vs 130-135) in number.

The present parasites differ from the species S. raoii Majid and Shinde,

1984 from Channa punctatus which is having scolex (tubular, cylindrical vs pear

shaped), hooks 46 in number, testes (350-370 vs 65-70) in number, vitellaria are

(follicular vs granular).

The present worm differs from the species S. jagannathae Majid and

Shinde, 1984 from Channa punctatus from India, which is having scolex

(tubular, cylindrical vs pear shaped), hooks 44 in number, testes (350-370 vs

240-250) in number, vitellaria are (follicular vs granular).

The present parasite differs from the species S. gachuae Jadhav et al.,

1991 from the host channa gachua in India which is having scolex (tubular,

cylindrical vs pear shaped), hooks (42-55 vs 22-25) in number, testes (350-370

vs 60-70) in number.

The present worm differs from the species S. maharashtrii Jadhav et al.,

1991 from Mastacembelus armatus in India, which is having scolex (tubular,

cylindrical vs oval), testes (350-370 vs 80-90) in number, vitellaria are follicular

with (single vs 4-5 rows).

The present parasite differs from the species S. chauhani Monzer Hasnain,

1992 from Channa punctatus in India which is having scolex (tubular, cylindrical

vs oval), hooks (42-55 vs 40-44) in number, neck (absent vs present); testes

(350-370 vs 200-210) in number, vitellaria are follicular with 4-5 rows.

The present worm differs from the species S. mohekarae Tat and Jadhav,

1997 from Mastacembelus armatus in India, which is having scolex (tubular,

cylindrical vs oval), hooks (42-55 vs 151) in number, neck ( absent vs long),

testes oval and (350-370 vs 300-310) in number, vitellaria follicular 3-4 rows in

each side.

The present worm differs from the species S. armatusae Hiware, 1999

from Mastacembelus armatus in India, in the presence of hooks (42-55 vs 32-40)

in number, mature proglottids four broader time than long, testes scattered, (350-

370 vs 230-240) in number, vitellaria two rows.

The present worm differs from S. tappi Patil et al., 2003 from

Mastacembelus armatus in India which is having scolex (tubular, cylindrical vs

triangular), testes (350-370 vs 285-295) in number.

The present parasite differs from the species S. ayodhensis Pande et al.,

2006 from Amphinuous cuchia in India, which is having scolex (tubular,

cylindrical vs conical), hooks (42-55 vs 29) in number, testes numerous.

The present worm differs from the species S. baught Pande et al., 2006

from Rita- rita in India. Which is having scolex (tubular, cylindrical vs pear

shaped), hooks (42-55 vs 28) in number, neck (absent vs present), and testes

(350-370 vs 40-50) in number.

The present worm differs from the species S. jadhavae Bhure et al., 2007

from Mastacembelus armatus which is having scolex (tubular, cylindrical vs

triangular), hooks 50-54 in number, testes oval (350-370 vs 120-150) in number.

The present worms differ from the species S. nathsagarensis Nilima M.

Kankale, 2008 from host Mastacembelus armatus which is having hooks are (42-

55 vs 30-32) in number, testes (350-370 vs 200-250) in number, vitellaria are

follicular in (single vs 2-3 rows).

Above distinct character are noted and justify the recognition of the

present worm as a new species and hence the name Senga rupchandensis n.sp. in

the honour of authors father Rupchand Khemchand Pardeshi.

Type species Senga rupchandensis n.sp.

Host Channa striatus (Bloch, 1793)

Habitat Intestine

Locality Godavari River, Shahagad, Jalna

District, M.S., India

Date of collection May 2007.

Senga rambaei n.sp.

DESCRIPTION:

Twelve cestode was collected from the intestine of Mastacembelus armatus

(Lecepede, 1800) from Jayakwadi dam, Paithan, Aurangabad district, M.S., India,

in the month of March 2007. Cestode were isolated from the intestine, washed in




cleared in xylene, mounting in DPX. Drawings were made with the help of


The worms were elongated, whitish in colour, segmented, strobila divided

into many immature, mature and gravid proglottids. Anterior portion are smaller

than posterior region. The total length of scolex 2.0908 mm. in length and 1.3067

mm. in breadth. The scolex is triangular in shape. Scolex bears two bothria which

extend upto posterior end of the scolex. Bothria spoon like, right bothria measures

1.4431 mm.in lengths and 0.3068 mm. in breadth, left bothria measures

1.5113mm. in length and 0.3409 mm. in breadth. Right bothria is smaller than left

bothria. The anterior part of the scolex ends terminally into large rostellum is

present and arranged with 47 semicircular hooks. The large hook measures

0.04171 mm. in length and 0.006417 mm. in breadth. Small hook measures

0.02245 mm. in length and 0.003208 mm in breadth. Neck is present; it measures

0.2727 mm. in length and 0.6704 mm. in breadth.

Mature proglottids longer than broad, it measures 1.6262 mm. in length and

0.2621 mm.in breadth. Genital pore on the cirrus pouch, it is oval or rounded in

shape, it measures 0.04368 mm. in diameter. Cirrus is small, thread like. Ootype is

the circular or rounded in shape, and located between two ovarian lobes .Cirrus

pouch is oval located at the middle portion of the proglottids, it measures 0.04854

mm. in length and 0.02912 mm in breadth. Testes are oval to rounded in shape,

having 120-150 in number. It measures 0.07766 mm. in diameter.

Ovary is bilobed, right lobe measures 0.04854 mm. in length and 0.05339

mm. in breadth and left lobe measures 0.1601 mm. in length and 0.07766 mm. in

breadth. Left ovarian lobe is larger than right lobe. Vagina is curved, tube like, and

connect to the Ootype, measures 0.2766 mm. in length and 0.03398 mm. in

breadth. Vitellaria follicular in single rows.

Gravid proglottids are longer than broad; it measures 1.6553 mm. in length

and 0.4029 mm. in breadth. Gravid proglottides broader than mature proglottids.

Eggs are oval, elongated in shape; it is operculated and measures 0.02245

mm. in length and 0.01176 mm. in breadth.

DISCUSSION:

The genus Senga was established by Dollfus, 1934, with its type species

S. bensardi from Betta splendens at Vincennes, France.

Later on described the twenty four species of Senga given are as follows,

1) S. ophicephalina Teseng, 1933 from Ophicephalus argus in Tsinan,

China.

2) S. bensardi Dollfus, 1934 from Betta splendeus in France.

3) S. pycnomera Woodland, 1934 from Ophiocephalus marulius in India.

4) S. lucknowensis Johri, 1956 from Mastacembelus armatus in India.

5) S. malayana Furnando and Furntado, 1964 from Channa striatus in

Malacca.

6) S. parva Furnando and Furtado, 1964 from Channa micropeltes in

Malacca.

7) S. pahanensis Furtado et al., 1971 from Channa micropeltis in Tasek,

Bera.

8) S. vishakhapatnamensis Ramadevi et al., 1973 from Ophiocephalus

punctatus in India.

9) S. khami Deshmukh and Shinde, 1980 from Ophiocephalus marulius in

India.

10) S. aurangabadensis Jadhav et al., 1980 from Mastacembelus armatus in

India.

11) S. godavari Shinde et al., 1980 from Mastacembelus armatus in India.

12) S. paithanensis Kadam et al., 1981 from Mastacembelus armatus in

Paithan, India.

13) S. raoii Majid M.A. and G.B. Shinde, 1984 from Channa punctatus in

India.

14) S. jagannathae M.A. Majid and G.B. Shinde, 1984 from Channa

punctatus in India.

15) S. gachuae Jadhav et al., 1991 from Mastacembelus armatus in Solapur

(M.S.), India.

16) S. maharashtrii, Jadhav et al., 1991 from Mastacembelus armatus in

Amravati, India.

17) S. chauhani Monzer Hashain, 1992 from Channa punctatus in

Jamshedpur (M.S.), India.

18) S. mohekarae Tat and Jadhav, 1997 from Mastacembelus armatus in

Osmanabad, India.

19) S. armatusae C.J. Hiware, 1999 from Mastacembelus armatus in Pune.

20) S. tappi D.N. Patil & B.V. Jadhav, 2003 from Mastacembelus armatus

in Shirpur, India.

21) S. ayodhenensis Pande et al., 2006 from Amphinuous cuchia in India.

22) S. baught pande et al., 2006 from Rita- rita in India.

23) S. jadhavae Bhure et al., 2007 from Mastacembelus armatus in India.

24) S. nathsagarensis Nilima M. Kankale, 2008 from Mastacembelus .

armatus in India.

25) S. rupchandensis n.sp. from Channa striatus.

The present worm under discussion is scolex triangular, two bothria, large

at the anterior end, hooks are semicircular, 47 in number, neck present, cirrus sac

oval, uterus coiled, tube like, genital pore oval and rounded, vagina tube like,

ootype circular, ovary bilobed, testes oval and 120-150 in number. Vitellaria are

follicular in single rows, eggs oval and operculated.

The present worm comes closer to the species. S. pahanensis Furtado et

al., 1971; S. paithanensis Kadam et al., 1981; S. tappi D.N. Patil, 2003 in having

scolex is triangular, anterior end pointed and posterior end broad, neck present,

cirrus pouch is oval, ovary bilobed, but it differs from S. pahanensis Furtado et

al., 1971 in having (47 vs 52) in number, Testes (120-150 vs testicular lobed),

vitellaria are (follicular vs lobate); S. paithanensis Kadam et al., 1981 having

hooks (47 vs 54), testes (120-150 vs 130 -155), vitellaria are follicular ( single vs

two rows); S. tappi D.N. Patil, 2003 having hooks (47 vs 42-44), testes (120-150

vs 285-295), vitellaria follicular to testicular. Remaining species differs from the

Senga rambaei n.sp.are as discussed below,

The present parasites differ from, the species S. ophicephalina Teseng,

1933 from Ophiocephalus argua in China which is having scolex (triangular vs

pear shaped), hooks 47-50 in number, neck is (present vs absent), testes are (120-

150 vs 50-55) in number, vitellaria (follicular vs lobate).

The present parasites differ from the species S. bensardi Dollfus,1934

from Betta splendens in France which is bothria shallow, hooks (47 vs 50) in

number, neck is (present vs absent), testes (120-150 vs 160-175) in number,

ovary is compact, not bilobed, vitellaria (follicular vs granular).

The present worm differs from the species S. pcynomera Woodland, 1934

from Ophiocephalus marulius in India which is having scolex (triangular vs

elongated), bothria shallow, hooks (47 vs 68) in number, neck is (present vs

absent), ovary discontinuous in two groups, vitellaria (follicular vs granular).

The present worm differ from the species S. lucknowensis Johri, 1956

from Mastacembelus armatus in India which is having scolex (triangular vs pear

shaped) with two shallow bothria, hooks (47 vs 36-48) in number, neck is

(present vs absent), in number of testes (120-150 vs 100-150), vitellaria (not

lobulate vs lobulate) and discontinuous two groups.

The present parasite differs from the species S. malayana Furnando and

Furtado, 1964 from Channa striatus in Malacca. Which is having scolex

(triangular vs circular), hooks (47 vs 60) in number, vitellaria lobate,

discontinuous in two groups.

The present parasites differ from the species S. parva Furnando and

Furtado, 1964 from Channa micropeltis in Malacca. Which is having scolex

(triangular vs pear shaped), hooks (47 vs 150-180) in number, testes (120-150 vs

100) in numbers, vitellaria are (follicular vs granular).

The present worms differ from the species S. visakhapatanamensis

Ramadevi et al., 1973 in having scolex (triangular vs circular), neck (present vs

absent), testes (120-150 vs 40-55) in number, vitellaria bilobed and post-

equatorial.

The present parasites differ from the species S. khami Deshmukh and

Shinde, 1980 from Ophiocephalus marulius in India which is having scolex

(triangular vs rectangular), hooks (47 vs 55-57) in number, neck is (present vs

absent), ovary is bilobed and post-equatorial, cirrus pouch is elongated, testes

(120-150 vs 155) in number.

The present parasite differs from the species S. aurangabadensis Jadhav et

al., 1980 from Mastacembelus armatus in India which is having scolex

(triangular vs oval), hooks (47 vs 50-52) in number, neck ( present vs absent),

ovary is bilobed and post-equatorial, testes (120-150 vs 240-260) in number,

vitellaria are corticular follicles.

The present worm differs from the species S. godavarii Shinde et al., 1980

from Mastacembelus armatus in having scolex (triangular vs pear shaped), hooks

(47 vs 40-42) in number, neck (present vs absent), testes (120-150 vs 230) in

number, ovary bilobed with short acini, vitellaria follicular 3-4 rows.

The present parasites differ from the species S. raoii Majid and Shinde,

1984 from Channa punctatus which is having scolex (triangular vs pear shaped),

hooks (47 vs 46) in number, neck (present vs absent), and testes (120-150 vs 65-

70) in number, vitellaria (follicular vs granular).

The present worms differ from the species S. jagannathae Majid and

Shinde, 1984 from Channa punctatus from India, which is having scolex

(triangular vs pear shaped), hooks (47 vs 44) in number, ovary bilobed, spatulate

and compact, testes (120-150 vs 240-250) in number, vitellaria (follicular vs

granular).

The present parasites differ from the species S. gachuae Jadhav et al.,

1991 from the host channa gachua in India, in the presence of scolex (triangular

vs pear shaped), hooks (47 vs 22-25) in number, neck (present vs absent), and

testes (120-150 vs 60-70) in number.

The present worms differ from the species S. maharashtrii Jadhav et al.,

1991 from Mastacembelus armatus in India, which is having scolex (triangular

vs oval), neck (present vs absent), hooks 22-47 in number, testes (120-150 vs 80-

90) in number, vitellaria follicular, rounded and 4-5 rows.

The present parasite differs from the species S. chauhani Monzer Hasnain,

1992 from Channa punctatus in India which is having scolex (triangular vs oval),

hook (47 vs 40-44) in number, neck is (present vs absent), testes (120-150 vs

200-210) in number, vitellaria non lobate to lobate.

The present worm differs from the species S. mohekarae Tat and Jadhav,

1997 from Mastacembelus armatus in India, which is having scolex (triangular

vs oval), hooks (47 vs 151) in number, neck long, testes oval and (120-150 vs

300-310) in number, vitellaria follicular 3-4 rows in each side.

The present worm differs from the species S. armatusae Hiware, 1999

from Mastacembelus armatus in India, which is having hooks (47 vs 32-40) in

number, neck ( present vs absent), mature proglottids four broader time than

long, testes scattered, (120-150 vs 230-240) in number .

The present parasite differs from the species S. ayodhenensis Pande et al.,

2006 from Amphinuous cuchia in India. Which is having scolex (triangular vs

conical), hooks (47 vs 29) in number, neck (present vs absent), testes numerous,

vitellaria small follicles.

The present worm differs from the species S. baught Pande et al., 2006

from Rita- rita in India. Which is having scolex (triangular vs pear shaped),

hooks (47 vs 28) in number, testes (120-150 vs 40-50) in number.

The present worm differs from the species S. jadhavae Bhure et al., 2007

from Mastacembelus armatus which is having hooks (47 vs 50-54) in number,

testes (120-150 vs 310-320), ovary is compact, oval, large coiled (bilobed vs

unilobed).

The present worms differ from the species S. nathsagarensis Nilima M.

Kankale, 2008 from host Mastacembelus armatus which is having scolex

(triangular vs long or elongated), hooks are (47 vs 30-32) in number, testes (120-

150 vs 200-250) in number, isthmus is short, vitellaria are follicular in (single vs

2-3 rows).

The present parasites differ from the species S. rupchandensis from

Channa striatus scolex is (triangular vs tubular, cylindrical), hooks (47

semicircular vs 42-55) in number, neck (present vs absent), testes oval or

rounded (120-150 vs 350-370) in number.

Above different character noted, justify the recognition of the present

worm as a new species and hence the name Senga rambaei n.sp. is proposed in

the honour of author mother Mrs. Rambai Rupchand Pardeshi who made him to

stand at this age.

Type species Senga rambaei n.sp.

Host Mastacembelus armatus

(Lecepede, 1800)

Habitat Intestine

Locality Jayakwadi dam, Paithan,

Aurangabad, District, (M.S.), India.

Date of collection March 2007.

Eucestoda Wardle, McLeod & Radinoky, 1974

Pseudophyllidea Carus, 1863

Ptychobothriidae Luhe, 1902

Circumoncobothrium Shinde, 1968


INTRODUCTION:

The genus Circumoncobothrium is erected by Shinde G.B., (1968)

described species C. Ophiocephalus from the intestine of freshwater fish,

Ophiocephalus leucopunctatus. Jadhav and Shinde, (1976) described three new

species i.e., C. aurangabadensis and C. raoii from the host Mastacembelus

armatus and C. gauchai from Ophiocephalus gauchua. Chincholkar and Shinde,

(1976) was reported two new species i.e., C. shindei from Mastacembelus

armatus and C. bagariusi from the freshwater fish, Channa striatus. Later,

Shinde added the new species C. khami in 1976 from the host Ophiocephalus

striatus. Jadhav et al., 1990 was reported new species C. yamaguti from

Mastacembelus armatus. Later Shinde et al., 1994 described new species C. alii

from the freshwater fish, Mastacembelus armatus. Later Patil et al., 1998

described new species C. vadgaonensis from host Mastacembelus armatus

Wongsawad and Jadhav, 1998 was added new species C. baimaii from

freshwater fish, Mastacembelus armatus. Kalse and Shinde, 1999 described two

new species C. punctatusi from host, Channa punctatus and C. armatusae

Shinde et al., 1999 from Mastacembelus armatus. Shinde et al., 2002 added a

new species C. mastacembelusae from the host Mastacembelus armatus. Later

Pawar et al., 2002 added new species C. armatusae (Minor) from Mastacembelus

armatus. Tat and Jadhav, 2004 described the new species C. manjari from the

fish Mastacembelus armatus. Supugade et al., 2005 described new species C.

vitellariensis from host Mastacembelus armatus. Later, described the new

species C. purnae by Borde S.N. and Sushil J. in 2008 from the host

Mastacembelus armatus.


Circumoncobothrium jadhavae n.sp. from Mastacembelus armatus (Lecepede,

1800).

DESCRIPTION:

Fifteen worms were recovered from the intestine of Mastacembelus

armatus (Lecepede, 1800) collected from Kham river at Aurangabad District,

Aurangabad (M.S.), India, in the month of April 2008. The worm were isolated

from the intestine, washed in distilled water, flattened between coverglass and

slides, fixed in 4% formalin until 24 hours, washed in distilled water, stained

with Harri’s Haematoxylene, dehydrated in ascending series of alcoholic grades

(30%,50%,70%,90%,100%), cleared in xylene and mounted in DPX. Drawings

were made by using Camera Lucida and measurements are taken in millimeter.

The worms are elongated, tape like, yellowish or cream in colour. The

scolex is triangular, dome shape and it measures 1.1022 mm. in length and

0.7727 mm. in breadth. Scolex bears two bothria, elongated, spoon like. Right

bothria measures 1.19311 mm. in length and 0.2045 mm. in breadth and left

bothria measures 0.9431 mm. in length and 0.1931 mm.in breadth. Hooks are

located on the tip of the scolex. Hooks are overlapping one another, 35-45 in

number; it measures 0.4812mm in length and 0.05347 mm. in breadth. Neck is

present.

Mature proglottids are longer than broad, measures 1.3688 mm. in length

and 0.3300 mm. in breadth. Cirrus pouch is oval, bulb like at the anterior end of

the vagina, measures 0.03398 mm in length and 0.01941 in breadth. Cirrus is

slightly curved at the anterior of genital opening, genital opening or pore is

small, rounded located on the cirrus pouch, and it measures 0.04368 mm. in

diameter. Testes are oval to rounded, 95-105 in number, and it measures 0.07281

mm in diameter.

Ovary is bilobed, right and left lobes separated from the ootype. Right

lobe measures 0.2330 mm. in length and 0.07766 mm. in breadth. Left lobe

measures 0.1213 mm. in length and 0.0484 mm. in breadth. Vagina is tube like at

the anterior of the genital pore measures 0.01456 mm. in length 0.004854 mm. in

breadth. Ootype is circular or rounded located between ovarian lobe and it

measures 0.04368 mm. in diameter. Isthmus at the distal portion of the ovary, it

measures 0.04368 mm in diameter. Mature proglottids are broader than gravid

proglottids. Vitellaria are follicular in two rows at the lateral side of the

proglottids.

Gravid proglottids longer than mature proglottids, it measures 1.4563 mm.

in length and 0.2281 mm. in breadth. Eggs are operculated and measures 0.02566


DISCUSSION:

The genus Circumoncobothrium is erected by Shinde G.B. (1968) with

species C. ophiocephali from the intestine of freshwater fish Ophiocephalus

leucopunctatus, in India. Later on eighteen species of this genus are added by

different authors which are as below

1. C. ophiocephali Shinde, 1968 from Ophiocephalus leucopunctatus in

India.

2. C. aurangabadensis Jadhav and Shinde, 1976 from M. armatus in India.

3. C. gachuai Jadhav and Shinde, 1976 from M. armatus in India.

4. C. raoii Jadhav and Shinde, 1976 from Mastacembelus armatus in India.

5. C. shindei Chincholkar and Shinde, 1976 from M. armatus in India.

6. C. bagariusi Chincholkar and Shinde, 1976 from Bagaricus sp. in India.

7. C. khami Shinde, 1976 in Ophiocephalus striatus in India.

8. C. yamaguti Jadhav et al., 1990 from Mastacembelus armatus in India.

9. C. alii Shinde et al., 1994 from Mastacembelus armatus in India.

10. C. vadgaonensis Patil, 1998 from Mastacembelus armatus in India.

11. C. baimaii Wongsawad and Jadhav, 1998 from Mastacembelus armatus in

India.

12. C. punctatusi Kalse and Shinde et al., 1999 from Mastacembelus armatus

in India.

13. C. armatusae Shinde et al., 1999 from Mastacembelus armatus in India.

14. C. mastacembelusaei Shinde et al., 2002 from Mastacembelus armatus in

India.

15. C. armatusae (Minor) Pawar et al., 2002 from Mastacembelus armatus in

India.

16. C. manjari Tat and Jadhav, 2004 from Mastacembelus armatus in India.

17. C. vitellariensis Supugage et al., 2005 from Mastacembelus armatus in

India.

18. C. purnae Borde S.N. and Sushil J., 2008 from the host, Mastacembelus

armatus in India.

The present parasite under discussion scolex is triangular, dome shape,

two bothria, and spoon like, hooks 35-45 in number, at the tips of the scolex and

overlap one another, neck is present, cirrus pouch small, bulb like, genital pore

small, rounded, ovary bilobed. Ootype circular, testes are oval or rounded, 95-

105 in number. Gravid proglottides longer than mature proglottids. Eggs are

operculated. Vitellaria are follicular in two rows at lateral side of the

proglottides.

The present parasite comes closer to C. vadgaonensis Patil, 1998; C.

armatusae (Minor) Pawar, 2002; C. manjari Tat and Jadhav, 2004 and C.

vitellariensis Supugale et al., 2005; in having scolex triangular, ovary bilobed,

mature segment is broader than long, vitellaria are follicular, however it differs

from C. vadgaonensis Patil, 1998 having hooks (35-45 vs 56) in number, testes

(95-105 vs 490-510) in number; C. armatusae (Minor) Pawar, 2002 in having

hooks (35-45 vs 58) in number, neck (present vs absent), testes (95-105 vs 190-

200) in number; C. manjari Tat and Jadhav, 2004 in having hooks (35-45 vs 48)

in number, testes (95-105 vs 128-145) in number; C. vitellariensis Supugale et

al., 2005 having hooks (35-45 vs 46-48 ), neck (present vs absent), testes (95-105

vs 250-260),above mentioned species vitellaria are follicular in two rows.

Remaining species differs from the Circumoncobothrium jadhavae n.sp.are as

discussed below,

The present worm differs from the species C. ophiocephali Shinde, 1968

from Ophiocephalus leucopunctatus in India in having scolex (triangular, dome

shape vs broad), hooks (35-45 vs 80) in number, testes (95 – 105 vs 70-80) in

number, ovary is (bilobed vs single conical mass to irregular shaped band).

The present worm differs from the species C. aurangabadensis Jadhav

and Shinde, 1976 from Mastacembelus armatus in having hooks 42 in number,

testes are (95-105 vs 135-145) in number, ovary is bilobed with 3-4 acini,

vitellaria are ( follicular vs granular).

The present worm differs from the species C. raoii Jadhav and Shinde,

1976 from Mastacembelus armatus in India. Which is having hooks (35-45 vs

46), broad in the middle and narrow at both ends, testes are (95-105 vs 210-215)

in number. Vitellaria are (follicular vs granular).

The present parasite differs from C. gachuai Jadhav and Shinde, 1976

from Mastacembelus armatus which is having scolex is (triangular, dome shape

vs pear shaped), hooks are (35-45 vs 46) in number, mature segments are

squarish, testes (95-105 vs 375-400) in number.

The present worm differs from the species C. shindei Chincholkar and

Shinde, 1976 from Mastacembelus armatus in India. Which is rostellar hooks

(35-45 vs 49) in number, testes are (95-105 vs 260-275) in number, vitellaria

(follicular vs granular).

The present parasites differ from the species C. bagariusi Chincholkar and

Shinde, 1976 from Bagarius sp. which is hooks (35-45 vs 55) in number, neck

(present vs absent) testes are (95-105 vs 275-285) in number.

The present parasite are differ from species the C. khami Shinde 1976 in

having scolex cylindrical, hooks (35-45 vs 48) in number, neck (present vs

absent), testes (95-105 vs 190-200) in number, mature segments is Squarish,

ovary bilobed, post-equatorial with short, blunt with 5-6 rows.

The present worms differ from species C. yamaguti Jadhav et al.,1990

from Mastacembelus armatus which is having hook (35-45 vs 56) in number,

neck is (present vs absent), testes are (95-105 vs 130-150) in number, vitellaria

are (follicular vs granular) .

The present worm differs from species C. alii Shinde et al.,1994 in having

hook (35-45 vs 34) in number, testes are (95-105 vs 230-240) in number,

vitellaria are (follicular vs granular).

The present worm differs from the species C. baimaii Wongswad and

Jadhav, 1998 in Ophiocephalus punctatus which is having scolex is (triangular vs

pear shaped), hooks (35-45 vs 48) in number, testes (95-105 vs 88-100) in

number, ovary (bilobed vs compact ), vitellaria (follicular vs granular) .

The present worms differ from the species C. punctatusi Kalse and

Shinde, 1999 from Mastacembelus armatus in having hooks 40-50 in number,

mature segment is squarish, 6-7 times broader than long, testes (95-105 vs 140-

150) in number, vitellaria (follicular, two rows at the lateral side vs follicular, 3-6

rows at the lateral side).

The present worms differ from the species C. armatusae Shinde et al.,

1999 from Mastacembelus armatus which is having hooks (35-45 vs 23) in

number, mature segment is 3-4 times broader than long, testes (95-105 vs 90-

100) in number, vitellaria (follicular, two rows at the lateral side vs follicular, 3-4

rows at the lateral side).

The present worm differs from the species C. mastacembellusaei Shinde,

et al., 2002 from Mastacembelus armatus which is having scolex (triangular vs

pear shaped), hook 38 in number, neck is (present vs absent), testes are small,

oval (95-105 vs 130-140) in number, ovary bilobed, compact.

The present parasites differ from the species C. purnae Borde S.N. and

Sushil J., 2008 from Mastacembelus armatus in having hooks (35-45 vs 52) in

number, mature segments squarish and broader than long, testes (95-105 vs 230-

235) in number, vitellaria follicular in 3-5 rows.

The distinct character as noted above, justify the recognition of the present

worm as a new species and hence the name Circumoncobothrium jadhavae n.sp.

is given in the memory of late Prof. Baba Jadhav who devoted his life for

promotion of Helminthology.

Type species Circumoncobothrium jadhavae n.sp.

Host Mastacembelus armatus

(Lecepede, 1800)

Habitat Intestine

Locality Kham river of Aurangabad

District, (M.S.), India

Date of collection April 2008.

Trematoda Rudolphi, 1808

Digenea van Beneden, 1858

Azygiidae Odhner, 1911

Azygiinae Luhe, 1909

Azygia Looss, 1899

Syn. Megadistomum Stafford, 1904†

Mimodistomum Stafford, 1904††

Hassallius Goldberger, 1911

Eurostomum MaCallum, 1921

Redescription Azygia stunkardi Rai, (1964)

INTRODUCTION:

The genus Azygia was erected by Looss, (1899) for accommodating

Fasciola lucii Mueller, 1776 syn. Azygia tereticollis Rud. (1802) from the

stomach of Erox lucius in Erope. Later described the species Azygia loossi by

Marshall & Gillbert, (1905) from Micropterus salmoides, Lucius lucius and Amia

calva; A. volgensis by von Linstow, (1907) from Luciperca Sandra; A. sebaga by

Ward, (1910) from Salmo sebago, A. bulbosa and A. acuminate by Goldberger,

(1911) from Amia calva. The family Azygidae was established by Odhner,

(1911) to contain the genera Azygia, Otodistomum, Leuceruthrus and

Ptychogonius. He was of the opinion that Magadistomum longum (Leidy, 1851)

from Exos ester, redescribed by Stafford, (1904), Mimodistomum angusticaudum

Stafford (1904) from Lota maculosa and Hassallius hassalli Goldberger, (1911)

from Ambloplites rupestris should be placed in the genus Azygia.

Odhner, (1911) also consider four species such as, A. loossi, A. balbosa,

A. angusticauuda, and A. acuminate to be syn. and added new species A. robusta

from Salmo hucho and Salmo fario. Cooper, (1915) described the species A. lucii

from Lucius lucius, L. masquiningy and Lucioperca sp. and he supported the

validity of A. acuminate. Later Fugita, (1918) reported the species A. perryi from

the host, Hucho perryi. Later Ozaki, (1924) described the species A. anguillae

from the host Angulla japonica in Japan. After, Manter in 1926 reviewing the

family Azygiidae recognized only three valid species according to him were

Azygia longa (Leidy, 1851) syn. (A. tereticolle, A. sebago, A. bulbosa, A. lucii

and Hassallius hassalli), A. angusticauda (Stafford, 1904) (syn. A. loossi) and A.

acuminate Goldberger, 1911. Later described the species such as, A. pritipomai

by Tubangui in 1928 from the host, Pristipoma hasta in the Philippines, A.

hwangtsinyi by Tsin in 1933 from the host, Ophiocephalus argus in China.

Zmeev, (1936) described the species A. amuriense from Ophiocephalus

argus in Russia. Later described the species life history of A. sebago by Stunkard

in 1956 and expressed the opinion that A. acuminate, A. bulbosa and Hassallius

hassalli may be identical with A. sebago. Stunkard considered A. angusticauda to

be valid and A. longa and A. lucii as distinct species. Later Velasquez, (1958)

considered the genera Eurostomum MacCallum 1921 and Gomtiotrema Gupta,

(1953) to be syn. with Azygia. Later added the species A. stunkardi Rai, 1964.

Jaiswal and Narayan in 1971 described new species A. marulii from

Ophiocephalus marulius.

The present communication deals with the description of redescribed

species Azygia stunkardi Rai, 1964, redescribed with some characters.

DESCRIPTION:

Ten worms were recovered from the stomach of Channa striatus (Bloch,

1793) collected from local market of Latur District, M.S., India, in the month of

May 2007. Trematode were collected or removed from the stomach, washed in




cleared in xylene, mounting in DPX. Drawings were made with the help of


The worms in the living condition were reddish in colour, showed

movements, contraction and expansion. The body of worm is elongated, non-

spinous, dorsoventrally flattened with round both ends (anterior and posterior

end). The entire worm measures 7.9545 mm. in length and 0.8636 mm. in breadth.

The oral sucker is oval, sub-terminal and located anterior end of the body. It

measures 0.4886 mm. in length and 0.4659 mm. in breadth. Prepharynx is absent.

Pharynx connects or behind the oral sucker and it measures 0.3636 mm. in length

and 0.2045 mm. in breadth. Oesophagus is short, sac like and infront of cirrus

pouch, measures 0.09090 mm. in length and 0.0568 mm. in breadth. The

acetabulum or ventral sucker is oval and intercaecal in position above the uterus; it

measures 0.4431 mm. in length and 0.4090 mm. in breadth. Oral sucker is larger

than ventral sucker. Intestinal caeca elongated and unequal arm.

In male reproductive organ, cirrus pouch is sac like, located between the

pharynx and oesophagus, it measures 0.6704 mm. in diameter. The vesicula

seminalis tubular, pars prostatica is coiled and opens into the vesicula seminalis.

They encloses in the thin walled of the cirrus pouch. Gonopore rounded,

intercaecal and above acetabulum. Testes entire, double, oval in shape located

below the Mehli’s gland or gland cells. Anterior testes oval and overlap to the

caeca, it measures 0.3181 mm. in length and 0.1818 mm. in breadth, above the

posterior testes. Posterior testes larger than anterior testes and overlap some part

of the ceca. Posterior testes measures 0.2727 mm. in length and 0.1931 mm, in

breadth.

Female reproductive organ, uterus is much coiled and overlap to the

intestinal caeca and some part of the ventral sucker. They locate in middle region

of the worm body. Vitelline follicles are numerus, rounded or oval, extracaecal in

position. Vitelline follicles are oval, double rows and start from below the ventral

sucker; it measures 0.126 mm. in length and 0.0795 mm. in breadth. Ovary is oval,

located at the posterior end of uterus and intercaecal in position; it measures

0.2727 mm. in length and 0.1931 mm. in breadth. Mehli’s gland or gland cell and

receptacle seminalis is located just infront of ovary and testes.

Excretory bladder absent. Eggs are elongated, non-operculated and

yellowish in colour. It measures 0.02887 mm. in length and 0.02352 mm. in

breadth.

DISCUSSION:

The genus Azygia was created by Looss in 1899 for accommodating

Fasciola loci Muller, 1776 syn. Azygia terticollis (Rudolph, 1802) from the

stomach of Esox lucius in Europe. The present species character such as

1) Oesophagus is short.

2) Intestinal caeca elongated and unequal arm.

3) Cirrus pouch located between the pharynx and oesophagus.

4) Vitelline follicles are double rows and start from below the ventral sucker

to unequal intestinal arm. This character resembles with species Azygia stunkardi

Rai, (1964) but having minor differ character such as,

1) Oesophagus notched.

2) Intestinal caeca is posterior extremity.

3) Cirrus sac preacetabular.

4) Vitelline follicles at posterior extremity.

Hence the species is redescribed with some additional characters as

mentioned above.

Type species Azygia stunkardi Rai, (1964)

redescribed species


Habitat Stomach

Locality Latur District, M.S., India

Date of collection May 2007.



Hemiuridae Luhe, 1901

Halipeginae Ejsmont, 1931

Genarchopsis Ozaki, 1925 syn. Progonus Looss, 1899;

Preoccupied Genarches Looss, 1902;

Preoccupied Ophiocorchis

Srivastava, 1933.

Genarchopsis paithanensis n.sp.

INTRODUCTION:

Looss, (1899) created the genus progonus to include Genarches mulleri

Levinsen. 1881. Ozaki, (1925) described the genus Genarchopsis for his

Genarchopsis gappo. Srivastava, (1933) synonymized Genarchopsis with

Progonus and described Progonus piscicola and Progonus ovocaudatum. In the

same year Srivastava created the genus Ophiocorchis to describe O. lobata and O.

singularis on account of the presence of oesophageal pouch. Gupta, (1951)

emended the diagnosis of the genus Ophiocorchis. Srivastava, (1933) and added

three more species O. dasus, O. indicus and O. faruquis. Yamaguti, (1958) taking

into consideration one common character the presence of caudal anastomosis in all

the genera-synonymised the genus Ophiocorchis Srivastava, (1933) (Genarches

Looss, 1902 preoccupied and Progonus Looss, 1899 preoccupied) with

Genarchopsis ozaki, 1925 retaining G. goppo as genotype.

He maintains following species under the genus Genarchopsis viz. G.

goppo Ozaki, 1925; G. dasus Gupta, 1951; G. lobata Srivastava, 1933; G. indicus

Gupta, 1951; G. faruquis Gupta, 1951; G. piscicola Srivastava, 1933 and G.

singularis Srivastava, 1933 all these species were reported from different fishes

.M. P. Dwivedi, (1965) described the species Genarchopsis melanostictus from

the stomach of Bufo melanostictus in Jabalpur, (M.P). Later Gupta S.P. and

Chakrabarti (1966) described the species G. thapari from intestine of snake in

Lucknow. Yamaguti, 1971 divided the genus Genarchopsis Ozaki, 1925 into two

subgenera, Genarchopsis (Ophiocorchis) with muscular metraterm and

Genarchopsis (Genarchopsis) without metraterm. Madhavi, R. (1971) described

the life history of Genarchopsis goppo Ozaki, 1925 recovered from Channa

punctatus. Bashirullah et al., (1972) described the species G. bangladensis and G.

ozaki from the intestine of the Channa punctatus in Bangladesh. Muley, (1972)

described the species of G. (Ophiocorchis) ozaki from intestine of Ophiocephalus

gachua in Aurangabad. The author (Bhadauria) collected new form of the genus

Genarchosis, subgenus Ophiocorchis obtained from stomach of Mastacembelus

armatus and Channa striatus. Later Bhadauria, (1984) described the species

Genarchopsis (Ophiocorchis) folliculata from the stomach of Mastacembelus

armatus and Channa punctatus, Gwalior (M.P.) and he has also described the

redescribed species as Genarchopsis (Genarchopsis) goppo Ozaki, (1925).

Chandra and Banerjee, (1993) described the new species Genarchopsis wallagoni

from the intestine of the Wallago attu in Bangladesh. Later Misako Urabe, (2001)

observe the life cycle of trematode Genarchopsis goppo.


Genarchopsis paithanensis sp. from the Mastacembelus armatus (Lecepede,

1800).

DESCRIPTION:

Seven worms were recovered from the intestine of Mastacembelus armatus

(Lecepede, 1800) from Jayakwadi Dam (Paithan), Aurangabad district, M.S.,

India, in the month of December 2007. Trematode were collected from the

intestine of fish, washed in distilled water, flattened between coverglass and

slides, fixed in 4% formalin until 24 hours, washed in distilled water, stained with

Harri’s Haematoxylene, dehydrated in ascending series of alcoholic grades (30%,

50% 70% 90% 100%), cleared in xylene, mounted in DPX. Drawings were made

with the help of Camera Lucida and all the measurement taken are in millimeter.

The worms are small, elongated and dorsoventrally flattened with rounded

anterior and posterior end, yellowish in colour, skin smooth and aspinous. The

entire worm measures 3.7954 mm. in length and 1.0227 mm. in breadth. The oral

sucker is sub terminal, oval in shape; it measures 0.2840 mm. in length and 0.2613

mm. in breadth. Ventral sucker measures 0.8409 mm. in length and 0.7954 mm. in

breadth. Oral sucker is smaller than ventral sucker. Pharynx at the distal part of

the oral sucker and it measures 0.06818 mm. in length and 0.1363 mm. in breadth.

Oesophagus opens into the pharynx and is tube like; it measures 0.0795 mm. in

length and 0.0454 mm. in breadth. The intestinal caeca is unite in front of

vitellaria.

The male reproductive organ, the cirrus pouch is well developed between

intestinal bifurcation it measures 0.9090 mm. in diameter. Vesicula seminalis is

curved; it measures 0.5568 mm. in diameter. Pars prostatica is tubular and opens

into the vesicula seminalis. Pars prostatica and hermaphroditic duct enclosed by

thin walled of cirrus pouch. The duct of pars prostatica and ejaculatory duct are

connected to the hermaphroditic duct. Testes are oval in shape and located below

the ventral sucker, embedded in the uterus. Right testes oval and measures 0.1818

mm. in length and 0.1590 mm. in breadth. Left testes just below the acetabulum,

rounded to oval above the ovary; it measures 0.1931 mm. in length and 0.1477

mm. in breadth.

Female reproductive organ, uterus is much coiled, filled with eggs and

around ventral sucker. Anterior region of the uterus filled with mature eggs and

located below and above the ventral sucker. Ovary is rounded at the posterior

region of the body and it measures 0.4886 mm. in diameter. Ovarian duct is tube

like and opens from the ovary and connect to the vitelline duct. Mehli’s gland oval

in shape below the ovary, embedded in the uterus and connect to the vitelline duct

and it measures 0.2159 mm. in length and 0.1704 mm. in breadth. Mehli’s duct

and ovarian duct are connecting together and go to the vitelline gland. Vitelline

gland is single, lobulated and located at the posterior region of the body and it

measures 0.6931 mm. in diameter.

Excretory bladder is absent. Eggs are oval, yellowish in colour and it

measures 0.03636 mm. in length and 0.1818 mm. in breadth.

DISCUSSION:

The genus Genarchopsis was erected by Ozaki in 1925 after Srivastava

(1933) created new genus Ophiocorchis, which differ from the Genarchopsis

only in having peculiar character termed by oesophageal pouch, but Yamaguti in

1958 considered Ophiochorchis a synonym of Genarchopsis.

1) O. indicus, (1951) from Ophiocephalus punctatus (Bloch.) in India.

2) O .dasus, (1951) from Ophiocephalus punctatus (Bloch.) in India.

3) O. faruquis, (1951) from Mastacembelus armatus in India.

4) O. lobatum (Srivastava, 1933) Yamaguti (1958), from Channa striatus

(Bloch.) in India.

Before 1958 some of the above author has described the species from genus as

Ophiocorchis but Yamaguti has synonymized the genus Ophiocorchis as

Genarchopsis in 1958 and later following species are added to this genus by

different authors are as below:

1) G. gappo Ozaki, 1925 from Mogurnda obscura in Japan.

2) G. lobata Srivastava, (1933) from Ophiocephalus striatus in India.

3) G. piscicola, Srivastava, (1933) from Channa punctatus in India.

4) G. singularis Srivastava, (1933) from Channa striatus in India.

5) G. dasus Gupta, (1951) from Channa punctatus, Glossogobium giuris in

Bangladesh

6) G. indicus Gupta, (1951) from Channa punctatus in India.

7) G. faruquis Gupta, (1951) from Mastecembelus armatus in India.

8) G. melanostictus Dwivedi, (1965) from stomach of Bufo melanostictus

in India.

9) G. thapari Gupta and Chakrabarti, (1966) from snake in Lucknow.

10) G. (Ophiocorchis) ozaki, Muley, (1972) from Ophiocephalus gachua in

Aurangabad, (M.S.), India.

11) G. bangladensis Bashirullah et al., (1972), from Ophiocephalus

punctatus (Bloch.) in Decca, Bangladesh.

12) G. (Genarchopsis) Ozaki Bashirullah et al., 1972 from Channa puctatus.

13) G. folliculata Bhadauria, 1984 from Mastacembelus armatus.

14) G. wallagoni Chandra et al., (1993), from intestine of Wallago attu

Bangladesh.

15) G. gappo Misako Urabe, 2001 from Semisulcospira libertine in Japan.

The present worm under discussion having oral sucker is sub-terminal,

oval, oral sucker smaller than ventral sucker, esophagus short, tube like, pharynx

well developed, intestinal caeca bifurcate and unite in front of vitelline gland,

cirrus pouch well developed, vesicula seminalis is curved, pars prostatica tubular,

uterus much coiled, ovary rounded, testes are oval, behind the acetabulum,

vitelline gland is single lobulate.

The present worm differs from the species G. gappo Ozaki 1925 from

Mogurnda obscura which is having the vesicula seminalis is (curved vs coiled),

vitellaria two, oval and clearly separate, excretory bladder (not vs Y shaped and

united anteriorly).

The present worm differs from the species G. lobata Srivastava, 1933

from Ophiocephalus striatus in having vesicula seminalis (curved vs bulbus),

anterior testis overlap to the ceca and posterior testis elongated, vitelline gland

(single, lobed vs two, 4-6 lobed).

The present worms differ from the species G. piscicola Srivastava, 1933

from Channa punctatus in having the oral sucker (0.28-0.26 vs x 0.33-0.34),

intestinal ceca (unite infront of vitelline gland vs attach to the vitelline gland),

uterus not extended to the vitellaria,

The present worms differ from the G. singularis Srivastava, 1933 from

Channa striatus in having the size of sucker (0.28-0.26 vs 0.23-0.47x0.37-0.81),

uterus not extended to the vitellaria.

The present worm differs from the species G. dasus Gupta, 1951 in having

oral sucker measures (0.28x 0.26 vs 0.23x0.37), ventral sucker (0.84x0.79 vs

0.62x0.55), and uterus not extended to the vitellaria.

The present worms differ from the species G. indicus Gupta, 1951

Channa punctatus in having intestinal caeca (unite infront of vitellaria vs broad

wavy), vesicula seminalis is (curved vs free from parenchyma), vitelline gland

are (single, lobulated vs two large lobed).

The present worms differ from the species G. faruquis Gupta, 1951 from

Mastacembelus armatus which is having oral sucker measures (0.28 x 0.26 vs

0.25 x 0.31), ventral sucker (0.84x0.79 vs 0.64x0.60), vitellaria are overlapped.

The present worms differ from the species G. melanostictus Dwivedi,

1965 from stomach of Bufo melanostictus in India. Which is having pharynx (not

overlapped vs overlapped) to oral sucker, testes (oval vs elongated), and uterus

(much coiled vs transverse convulated), excretory bladder (not found vs Y

shaped).

The present worms differ from the species G. thapari Gupta and

Chakrabarti, 1966 from the intestine of snake in Lucknow, which is having

oesophagus pouch (tube like vs small), vitellaria (single, lobulated vs two lobed),

excretory bladder (not found vs Y-shaped).

The present worm differs from the species G. (Ophiochorchis) ozaki,

Muley, 1972 from intestine of Ophiocephalus gachua in Aurangabad, in the

presence of pharynx (muscular vs globular), nipple shaped genital papillae,

excretory bladder (not found vs Y-shaped).

The present parasites differ from the species G. bangladensis Bashirullah

et al., 1972 from Ophiocephalus punctatus (Bloch.) in Decca, Bangladesh. This

is having intestinal caeca connect to the vitellaria, testes (oval vs asymmetrical

rounded), left testes elongated and right testes spherical, excretory bladder Y

shaped.

The present worms differ from the redescribed species G. ozaki

Bashirullah et al., 1972 from Mastacembelus armatus having intestinal caeca

(unite in front of vitellaria vs posterior extremity), testes (oval vs asymmetrical)

and vitelline gland (single, lobulated vs two, compact).

The present parasite differs from the species G. folliculata Bhadauria et

al., 1984 which is having esophagus (tube like vs short), vesicula seminalis

(curved vs tubular), and vitellaria are (single lobulated vs five to ten follicles),

excretory bladder is tubular.

The present parasites differ from the species G. wallagoni Chandra et al.,

1993 from intestine of Wallago attu in Bangladesh. Which is having intestinal

caeca (short vs broad and wavy), testes (oval vs asymmetrical), vesicula

seminalis (curved vs elongated coiled tube), and vitelline gland (single lobulated

vs two large) compact bodies.

The present parasites differ from the species G. goppo (Ozaki, 1925),

Misake Urabe, 2001 redescribed species which is having cirrus sac is absent,

vitellaria (single, lobulates vs two rounded or lobed), excretory bladder tubular.

All above differentiating characters are noted, justify the erection of this

present worm as a new species and hence the name Genarchopsis paithanensis

n.sp. is proposed after the locality Paithan from which the host collected.

Type species Genarchopsis paithanensis n. sp

Host Mastacembelus armatus (Lecepede, 1800)

Habitat Intestine

Locality Jayakwadi Dam (Paithan),

Aurangabad District, M.S., India

Date of collection Dec. 2007.



Gorgoderidae Looss, 1901

Phyllodistominae (Nybelin, 1926) Yamaguti, 1958

Phyllodistomum Braun, 1899

Phyllodistomum aurangabadensis n.sp.

INTRODUCTION:

Looss, (1901) erected the family Gorgoderidae with two subfamilies,

Gorgoderinae and Anaporrhutinae and under the latter, genus included

Anaporrhutum oftenheim, (1900); Probotitrema Looss, 1901 and Plesiochorus

Looss, 1901. Subsequently several genera added Petaldistomum Johnston, 1912,

Staphylorchis Travassos, 1926; Dendrochis Travassos, 1926 and Nagmia

Nagaty, 1930. In his review on sub-family Anaporrhutinae. Nagaty, (1930)

removed Dendrorchis from the sub-family and fused it with the genus

Phyllodistomum under Gorgoderinae, Looss, (1902), placed under the subfamily

Goroderinae, Phyllodistomum Braun, 1899 syn. Spathidium Looss, 1899,

Gordera Looss, 1902. Linton, (1910) added the genus Xystrum and Travassos,

(1920) erected the genus Macia from Cataptroides magnum and Cataptroides

aluterae MacCallum, 1917. Ozaki, 1926 added Microlecithus but Yamaguti,

(1934) and Bhalerao, (1937) regarded it syn. to Phyllodistomum. Lewis, (1935),

Lynch, (1936) and Bhalerao, (1937) disregarded the genus Cataptroides and

placed all its species under the genus Phyllodistomum. Pande, (1937) considered

the genus Gorgoderinae also synonym to Phyllodistomum.

Phyllodistomum Braun, 1899 have been described the species P.

spatulaeforme Odhner, 1902; P. spatula odhner, 1902 from African freshwater

fishes, catfishes. Looss, 1899 describe the species Phyllodistomum acceptum.

Later Yamaguti, 1934 described the species P. parasiluri; Bhalerao, 1937

described the species P. shandrai Pande, 1937 described the species P. almorai.

Later Steelman M. Gerald described the species P. caudatum in 1938 from

bullhead in Oklahoma. Jagdeshwari and Dayal, 1949 described the species P.

vachius from urinary bladder of Eutroplicthys vacha (Ham.) in Lucknow. Later

Kaw, 1950 added three species; P. loosi, P. frequentum, Phyllodistomum species,

Jacob H. Fischthal, 1942, added the three species such as P. semotili from

urinary bladder of Semotilus atropidus (Mitchill) in Michigan; P. notropidus

from urinary bladder of Notropis cornatus, Chrysocephalus; P. nocomic from

Nocomis biguttatus in Michigan.

Gupta, 1951 added the species P. singhai from intestine of Mastacembelus

armatus in Lucknow; Fischthal, (1952) described the species P. lysteri from

ireters and urinary bladder of Catostomus commersonnii; Motwani et al., (1961)

described the two new species P. chauhani from urinary bladder of Mystus

(Osteobagarus) aor (Ham), Mystus seenghala in Allahabad (U.P.) and P.

tripathii from Bagarius bagarius (Ham.).

Kenneth et al., 1982 added redescribed the species P. americanum

Osborn, 1903 from urinary bladder of Ambystoma tigrinum tigrinum; Wang

Xiyum, (1984) described the two species such as P. hemiculteri from Hemiculter

leucisculus in Poyang Lake, Jianxi province. ; P. plagiognathops from urinary

bladder of plagiognathops in Jiangxi Province.

Zhang, (1991) described the three new species P. saurogobia from

excretory bladder of Saurogobio dadrye (Bleeker) in China. P. leiocassis from

excretory bladder of freshwater fish, Leiocassis macrops in Fuzhou, P.

congrenensis from excretory bladder of Leiocassis crassilaris in Pinxiang. Later

Suzana et al., (1993) added the species P. rhamdiae from catfishes (R. queens) in

Brazil.


Phyllodistomum aurangabadensis n.sp. from Channa striatus (1793).

DESCRIPTION:

Two worms were recovered from the intestine of Channa striatus (Bloch,

1793) from Kham River, Aurangabad district, M.S., India, in the month of

December 2008. Trematode were collected or removed, washed in distilled water,

flattened between coverglass and slides, fixed in 4% formalin until 24 hours,

washed in distilled water, stained with Harri’s Haematoxylene, dehydrated in

ascending series of alcoholic grades (30%, 50% 70% 90% 100%), cleared in

xylene, mounted in DPX. Drawings were made with the help of Camera Lucida

and all the measurement are taken in millimeter.

The worms are small, posterior part is saucer shaped, smooth and reddish in

colour. The entire worm measures 3.6362 mm. in length and 2.9431 mm. in

breadth the oral sucker oval, terminal in position. Oral sucker measures 0.2613

mm. in length and 0.1477 mm. in breadth. Oral sucker larger than ventral sucker.

Pharynx is absent. Oesophagus tube like, below the oral sucker, it measures

0.1704 mm. in length and 0.04545 mm. in breadth. Intestinal caeca elongated,

broad, bifurcate and posterior extremity. Gonopore oval or rounded, between the

intestinal bifurcations. Acetabulum oval below the gonopore and it measures

0.2045 mm. in length and 0.1818 mm. in breadth.

Male reproductive organ, cirrus pouch well developed. The pars prostatica

pouch is sac like; it measures 0.1704 mm. in diameter. The vesicula seminalis is

small, curved and opens from the pars prostatica and it measures 0.1022 mm. in

length and 0.03409 mm. in breadth. The hermaphrodite duct is elongated, zigzag

in manner; it measures 0.1704 mm. in length and 0.03409 mm. in breadth, it opens

into the genital opening and pars prostatica pouch. Genital pore small, oval it

measures 0.05681 mm. in length and 0.02272 mm. in breadth. Prostatica cell and

ejaculatory duct are enclosed by thin walled of cirrus pouch. Testes are large,

double. Left testes are large, rectangular in shape, below the ovary, intercaecal in

position and it measures 0.4772 mm. in length and 0.3968 mm. in breadth. A right

testis is transversely elongated, oval in shape below the vitelline gland. The right

testes smaller than left testes and overlap to the intestinal caeca, measures 0.3977


Female reproductive organ, ovary transversely oval, intercaecal in position

and measures 0.2727 mm.in length and 0.1931 mm. in breadth. Ovary locates just

side of the vitelline gland. Vitelline glands bilobed, butterfly shaped. Left lobe

measures 0.3068 mm. in length and 0.1136 mm. in breadth .right lobe measures

0.2954 mm. in length and 0.1363 mm. in breadth. Vitelline gland below the

acetabulum. Uterus has numerous coils, attached to the vitelline gland, excretory

pore small between the intestinal extremities.

Eggs are numerous, rounded or oval in shape, non- operculated and it


DISCUSSION:

Phyllodistomum Braun, 1899 have been described the species P.

spatulaeformae Odhner, 1902; P. spatula Odhner, 1902 from the African

freshwater fishes, catfishes. Later present parasites are discussed with the earlier

known species of the genus Phyllodistomum.

1) P. spatuaeforme Odhner, 1902 from Ophicephalus punctatus in Africa.

2) P. spatula Odhner, 1902 from Bagrus bayad in Africa.

3) P. parasiluri Yamaguti, 1934 from Amyda sinensis in South Korea.

4) P. almorai Pande, 1937 from Rana cyanophlyctis in Kumaon hills.

5) P. shandrai Bhalerao, 1937 from Rana tigrina in Bombay.

6) P. caudatum Steelman, 1938 from Bullhead in Oklahoma.

7) P. semolitis Fischthal, 1942 in Semolitis atromaculatus (Mitchill),

Michigan.

8) P. notropidus Jacob H. Fischthal, 1942 from Notropis cornutus

chrysocephalus in Michigan.

9) P. nocomis Fischthal, 1942 from Nocomis biguttatus (Rafinesque),

Michigoan.

10) P. vachius Dayal, 1949 from Eutropiicthys vacha (Ham.) in Lucknow.

11) P. loossi Kaw, 1950 from Bufo viridis in Kashmir.

12) P. frequentum Kaw, 1950 from Bufo viridis in Kashmir.

13) P. singhai Gupta, 1951 from Mastacembelus armatus in lucknow.

14) P. lyseri (Miller. 1940) Fischthal, 1952 from Catostomus commersoni in

Michigan.

15) P. chauhani Motwani et al., 1961 from Mystus (Osteobagarus) aor

(Ham), Mystus seenghala in Allahabad.

16) P. tripathii Motwani et al., 1961 Bagarius bagarius in Allahabad.

17) P. americanum (Osborn, 1903) redescribed from Ambystoma tigrinum

tigrinum Kennath et al., 1982 in Wisconsin

18) P. hemiculteri Wang Xiyum, 1984 from Hemiculter leucisculus in Jianxi

Province.

19) P. plagiognathops Wang Xiyum, 1984 from Plagiognathops Jianxi

Province.

20) P. saurigobia Zhang, 1991 from Saurogobio dadrye (Bleeker) in

Nanchang.

21) P. leiocassis Zhang, 1991 from Leiocassis macrops in Fuzhou.

22) P. congrenensis Zhang, 1991 from Leiocassis crassilaris in Pinxiang.

23) P. rhamdiae Suzana et al., 1993 from catfishes (R. quelen) in Brazil.

The present worm under discussion having oral sucker is oval, terminal,

oral sucker larger than ventral sucker, esophagus tube like, intestinal caeca

elongated, broad and posterior extremity, acetabulum is rounded, cirrus pouch

well developed, vesicula seminalis is small, curved, ovary transversely oval,

testes are oval, rectangular, vitelline gland is bilobed or butterfly like.

The present worms closer to P. parasiluri Yamaguti, 1934; P. chauhani

Motwani et al., 1961; P. tripathi Motwani et al., 1961; which is having oral

sucker is terminal, esophagus is present, intestinal caeca broad and posterior

extremity, vesicula seminalis is sac like, vitelline gland is paired however it

differs from the species P. parasiluri Yamaguti, 1934 in having testes (oval to

rectangular vs deeply multilobed), ovary (oval vs multilobed); P. chauhani

Motwani et al., 1961 having testes (oval to rectangular vs slightly pear shaped);

P. tripathi Motwani et al., 1961 which is having testes (oval to rectangular vs 4-6

lobe), ovary (oval vs combed) and above mentioned species, cirrus sac (present

vs absent). Remaining species differs from the phyllodistomum aurangabadensis

n.sp.are as discussed below,

The present worms differ from the species P. spatulaeforme Odhner, 1902

which is having oral sucker sub terminal, cirrus sac ( present vs absent), vesicula

seminalis (curved vs sac like free from the parenchyma), excretory bladder

tubular.

The present worm differs from the species P. spatula Odhner, 1902 in

having oral sucker rounded, cirrus pouch oval, testes two and 4-5 lobe, ceca

bifurcate and broad.

The present worms differ from the species P. almorai, 1937 from Rana

cyanophlyctis in Kumaon hills. Which is having ventral sucker (larger than oral

sucker vs smaller than oral sucker), testes diagonal, may be deeply (without

notched vs notched), ovary is (oval vs slightly lobed), vitelline gland (lobed or

butterfly like vs compact).

The present worm is differing from the species P. shandrai Bhalerao,

1937 from Rana tigrina in Bombay. Which is having vesicula seminalis is

(small, curved vs large, bipartite).

The present worm differs from the species P. caudatum Steelman, 1938

from Bullhead which is having oral sucker (oval vs circular), oesophagus (tube

like vs sinous), ovary (oval vs 3-6 lobes), and testes (oval or rectangular vs

compact).

The present worm differs from the species P. simolitis Fischthal, 1942

from the host Semolitis atromaculatus (Mitchill),Michigan. which is having

oesophagus weekly muscular, slender, narrow, ovary compact (oval vs slightly

lobate), testes (oval or rectangular vs lobate), vitelline gland (lobed or butterfly

like vs oval), excretory bladder tube like.

The present worm differs from the species P. notropidus Fischthal, 1942

from Notropis cornutus chrysocephalus in Michigan in the presence of

esophagus (tube lile vs weakly developed), seminal vesicle (small, curved vs

large), ovary (oval vs compact), testes (oval or rectangular vs irregularly lobate),

excretory bladder slender.

The present worm differs from the species P. nocomis Fischthal, 1942

from Nocomis biguttatus (Rafinesque), Michigoan. Which is having oesophagus

(tube like vs slender and narrow), vesicula seminalis (small, curved vs large),

ovary (oval vs compact), vitelline gland (lobed, butterfly like vs oval), excretory

bladder slender.

The present worm differs from the species P. vachius Dayal, 1949 from

Eutropiicthys vacha (Ham.) in Lucknow. which is having oesophagus small and

forms a loop before caeca bifurcate, caeca simple, vesicula seminalis (small,

curved, not free vs free from parenchyma), ovary (oval,without lobe vs 5-6

lobed), testes two (oval or rectangular vs triangular), vitelline gland (butterfly

like vs undivided two follicles), excretory bladder tubular.

The present worm differs from the species P. loossi, 1950 from Bufo

viridis in Kashmir which is having oesophagus narrow, vesicula seminalis

(small, curved vs bipartite), ovary ( oval vs lobulated), and testes (oval or

rectangular vs diagonal).

The present worm differs from the species P. frequentum 1950, from Bufo

viridis in Kashmir. Which is having vesicula seminalis (small, curved vs large

bulbous), ovary (oval vs pear shaped).

The present worm differing from the species P. singhai Gupta, 1951 from

Mastacembelus armatus in Lucknow, which is having esophagus leads directly

to the caeca, vesicula seminalis (small, curved, not free vs free from the

parenchyma), ovary (without lobe, oval vs 4-5 lobed), testes (oval or rectangular

vs 6 or 7 lobes), vitelline gland (lobed, butterfly like vs two undivided lobes),

excretory bladder tubular.

The present worm differs from the species P. lyseri Fischthal, 1952 from

Catostomus commersoni in Michigan. Which is having ventral sucker (larger

than oral sucker vs smaller than oral sucker), ovary (oval vs compact), and testes

(oval or rectangular vs lobulate).

The present worm differs from the species P. americanum (Osborn, 1903)

Kennath et al., 1982 from Ambystoma tigrinum tigrinum (Green) in Wisconsin,

in the presence of ovary (oval vs irregularly to deeply lobed), testes (tandem,

oval or rectangular vs slightly oblique and deeply lobed).

The present worms differ from the species P. hemiculteri from Hemiculter

leucisculus which is having ovary (oval vs three lobed), and P. plagiognathops

Wang Xiyum, 1984 from Plagiognathops in Jianxi Province, in the presence of

ovary (oval vs Multilobed transversely and longitudinally slightly lobed). Above

both species having testes two (oval or rectangular vs diagonal in shape or

slightly lobed), vitelline gland (lobed, butterfly like vs oval), excretory bladder

tubular.

The present worm differs from the species P. saurigobia Zhang, 1991

from Saurogobio dadrye (Bleeker) in Nanchang, in the presence of ovary (oval

vs anterior notched), testes (oval or rectangular vs lobulated), excretory bladder

tubular.

The present worms differ from the species P. leiocassis Zhang, 1991 from

Leiocassis macrops in Fuzhou, in the presence of ventral sucker larger than oral

sucker, caeca (broad vs zigzag in manner), ovary (oval vs slightly lobed), testes

(oval or rectangular vs 9-12 lobed), vitelline gland (lobed, butterfly like vs

undefinately lobed).

The present worms differ from the species P. congrenensis Zhang, 1991

from Leiocassis crassilaris in Pinxiang, which is having oral sucker smaller than

ventral sucker, esophagus (tube like vs bifurcate), testes (oval or rectangular vs

4-5 lobed), vitelline gland (butterfly like vs oval and bilobed).

The present worms differ from the species P. rhamdiae Suzana et al.,

1993 from catfishes (R. quelen) in Brazil. Which is having seminal vesicle

(small, curved vs saccute) and preacetabular, ovary (oval vs lobulated), testes

(oval or rectangular vs lobed and slightly oblique).

Phyllodistomum aurangabadensis n.sp. named after the locality

Aurangabad, M.S. India where the Dr. Babasaheb Ambedkar Marathwada

University is located and author is working for his Ph.D. degree.

Type species Phyllodistomum aurangabadensis n.sp.


Habitat Intestine

Locality Kham River, Aurangabad District,

M.S., India

Date of collection Dec. 2008.



Allocreadiidae (Looss, 1902) Stossich, 1903

Allocreadiinae Looss, 1902

Allocreadium Looss, 1900

Allocreadium khami n.sp.

INTRODUCTION:

Allocreadium was erected by Looss in 1894, 1900 with A. isoporum as its

species. The name Creadium given to a genus of birds by Viellot in 1916, being

very similar to Creadium and according to the usual view precluding the use of the

latter term, Looss, (1900) changed the name to Allocreadium.

Stossich, (1900) placed the following species in the genus Allocreadium

such as A. pegorchia Stoss. A. obovatum (Molin) and A. asymphyloporum Stoss.

Odhner, (1901) revised the genus Allocreadium placing the following species A.

fascinatum (Rud.), A. labri (Stoss.). Later, 1901 found it to be synonymous with

A. pulchella (Rud.). A. tubibulum (Rud.), A. labracis (Duj.), A. commune (Olss.),

and A. genu (Rud.).

Later, in (1902) Odhner placed A. fasciatum, A. pulchella and A. sinuatum

in new genus Helicometra. Stafford, (1904) catalogues Allocreadium isoporum

Looss from the intestine of Semotilus bullaris. Later described the new species A.

lobatum by Wallin E. Ivan in (1909) from the Semotilus bullaris, A. colligatum, A.

mormyri (Stoss.), A. pallen (Rud.).

Odhner, (1928) and Dollfus, (1937) had already removed A. annandali

Southwell, 1913 to the Azygiidae. First record of the genus Allocreadium in India

from freshwater fishes are made by Pande, 1937. Pande described the new species

A. handiai from Ophiocephalus punctatus and Rita-rita. In 1950 Kaw, described

the new species A. nemacheilus from Nemacheilus kashmirensis. Later Gupta,

1950 described the new species A. thapari from freshwater fish, Rita-rita. In 1956

Gupta described two new species A. mehrai and A. kamlai from Rhynchobdella

aculeate and Chela bacaila respectively. Yamaguti, (1953) has made it more

inclusive by reducing Cainocreadium, Paracreadium and Lepidauchen to the rank

of subgenera in Allocreadium. Species thereby added to the genus are rejected by

the author, thus leaving in addition to A. neotenicum, the following species for

further evaluation such as A. symphyloporum Stossich, 1900; A. bolesomi Pearse,

1924; A. chuscoi Pearse, 1920; A. dogieli Koval, 1950; A. dubium Stossich. 1905;

A. fowleri Leiper et Atkinson, 1914; A. handiai Pande, 1937; A. hasu Ozaki, 1926;

A. kosia Pande, 1938a; A. lobatum Wallin, 1909; A. maheseri Pande, 1938b; A.

markewitschi Koval, 1949; A. mormyri (stossich); A. nemachilus Kaw, 1950: A.

nicolli Pande, 1938a; A. obovatum (Molin, 1858); A. oncorhynchi Eguchi, 1931;

A. pallens (Rud.) 1819; A. schizothoracis Pande, 1938b; A. striatum Dinulescu,

1941; A. thapari Gupta, 1950; A. transvarsale (Rud.) 1802; A. umbrinae (stoss,

1885); and A. Wallini Pearse, 1920.

Srivastava, 1960 added the new species A. ophiocephali from the

Ophiocephalus punctatus. Rai, 1962 has described species A. dollfusi; A. singhi

and A. hirnai all from the host Barbus tor. Gupta, 1963 added new species A.

makundi from Barbus sarana. Agarwal, 1964 described the new species A.

hetropneuatusius from Heteropneustes fossilis.

Mehra, (1966) revised the classification of Allocreadium Nicoll, 1935 and

recognized the following valid species under the genus Allocreadium namely A.

handiai Pande, 1937 syn. of A. thapari Gupta, 1950; A. ophiocephali Srivastava,

1960; A. nemachilus Kaw, 1950; A. mehrai Gupta, 1956 and A. kamlai Gupta,

1956.

Kakaji, (1969) disagreed with Mehra, (1966) who considered that A.

thapari to be syn. of A. handiai. A. thapari is distinct from A. handiai in the

position of testes and receptaculum seminalis and relative size of eggs. Kakaji,

(1969) considered following valid species in Allocreadium genus i.e., A. handiai

Pande, 1937; A. thapari Gupta, 1950; A. mehrai Gupta, 1956; A. singhi Gupta Rai,

1962; A. heteropneutusius Agarwal, 1964; A. dollfusi Rai, 1962; A. hirnai Rai,

1962; A nemachilus Kaw, 1950; A. makundi Gupta. 1963; A. kamlai Gupta, 1956.

Rai, 1970 agreed with Mehra, 1966 and redescribed A. mehrai Gupta, 1956

and A. handiai Pande, 1937 from freshwater fishes. Thomas J.D., (1972) described

the species A. ghanensis from rectum of Synodontis sp. (Mochocidae) in Ghana.

Gupta and Verma, 1976 described three new species such as A. mrigala, A.

saranai and A. baranai from the host, Barbus sarana and Barilius barna

respectively. Madhavi and Hanumantha Rao, 1977 described the anatomy of the

female reproductive system in A. ophiocephali Srivastava, 1960. Later Madhavi in

1978 worked out the life history of A. fasciatusi Kakaji, 1969 from freshwater

fish, Aplocheilus malastigma and observed variations in egg size, shape of

excretory bladder structure of seminal vesicle and presence of eye spot.

In 1980, Madhavi described the species A. handiai Pande, 1937 from

Channa punctatus. Bhadauri, 1984 described the two new species A. gwaliorensis

and A. tigarai from the intestine of freshwater fish, Mystus vittatus and Channa

punctatus respectively. Later, added A. gotoi (Hasegawa and Ozaki, 1926)

Shimazu in 1988 from the intestine of Misgurnus anguolocaudatum in Oshima.

Shimazu, (1988) described the species A. tosai from the Morcopercnurua

sachalinensis (cyprinidae) in Kushiro.

Later Ernest et al., 1993 described new species A. lucyae from bandfin

shiner, Notropis zonistius in Illinosis. Takeshi et al., (2000) described the species

A. patagonicum from the intestine of Percichythys colhuapiensis in Argentina.

Takeshi Shimazu, (2003) described the two species such as, A. shinanoense and A.

aburahaya from intestine of Phoxins lagowskii steindacheri sauvage in Japan.

The present communication deals with description of new species

Allocreadium khami n.sp. from the Mastacembelus armatus (Lecepede, 1800).

DESCRIPTION:

Ten worms were recovered from the liver of Mastacembelus armatus

(Lecepede, 1800) from Kham river, Aurangabad district (M.S.), India, in the

month of December 2008. Trematode were collected from the liver of fish,

washed in distilled water, flattened between coverglass and slides, fixed in 4%

formalin until 24 hours, trematode worms after removing from preservatives,

washed in distilled water, stained with Harri’s Haematoxylene, dehydrated in

ascending series of alcoholic grades (30%, 50% 70% 90% 100%), cleared in

xylene, mounted in DPX. Drawings were made with the help of Camera Lucida

and all the measurement taken are in millimeter.

The trematode worm are large, dorsoventrally flattened, reddish in colour,

skin smooth, the entire length of worm measures 24.8213 mm. in length and

10.6249 mm. in breadth. The oral sucker is rounded, sub terminal, it measures

3.2857 mm. in diameter. Pharynx is oval and attached posterior side of the oral

sucker, esophagus not visible. Ventral sucker oval, larger than oral sucker below

the cirrus pouch and gonopore it measures 1.6607 mm. in length and 1.3392 mm.

in breadth. Intestinal caeca elongated, bifurcate, bulging, overlap to the ovary and

uterus and posterior extremity. Gonopore small, rounded between the esophagus

and ventral a sucker.

Male reproductive organ, cirrus pouch is well developed, elongated, spoon

like and located side or above ventral sucker and overlapping to the intestinal

caeca, it measures 1.9464 mm. in diameter. Cirrus is long tube like; anterior of the

genital opening and it measures 0.2678 mm. in length and 0.05357 mm. in

breadth. Genital opening is oval and it measures 0.07142 mm. in length and

0.05357 mm. in breadth. Ejaculatory duct is tubular at the anterior end of the pars

prostatica and measures 0.2857 mm. in length and 0.03571 mm. in breadth. Pars

prostatica cells enclosed by thin walled of the cirrus pouch. Pars prostatica is tube

like, zigzag manner and opens into the seminal vesicle. Vesicula seminalis oval,

bipartite, it measures 0.1954 mm. in length and 0.1071 mm. in breadth. Testes are

two, circular at the posterior side of the body. Anterior testes free from the uterus

and measures 1.5178 mm. in diameter.

Female reproductive system, uterus is large or much coiled, broad and

overlaps to the ventral sucker and posterior testes. Ovary is longitudinally slightly

oval, large and it measures 1.3121 mm. in length and 1.010 mm. in breadth.

Vitellaria are confluent or branched like at the posterior region of the body.

Excretory bladder is tubular.

Eggs are elongated and it measures 0.05719 mm. in length and 0.02772

mm. in breadth.

DISCUSSION:

Allocreadium was created by Looss in 1894, 1900 with A. isoporum as its

species. The following known species of Allocreadium has been added by

different workers.

1) A. isoporum Looss, 1894, 1900 from Tribolodon ezoe in Hakkaido.

2) A. lobatum Wallin E. Ivan, 1909 from the Semotilus bullaris in Nebraska.

3) A. nemachilus Kaw, 1950 from the Schiozothoran micropogan in Kashmir.

4) A. thapari Gupta, 1950 from host Eutropiichthys vacha in India.

5) A. kamlai Gupta, 1956 from Chela bacaila (Ham.) in India.

6) A. mehrai Gupta, 1956 from Rhynochobella oculeata (Bloch.) in India.

7) A. ophiocephali Srivastava, 1960 from Ophiocephalus punctatus in India.

8) A. dollfusi Rai, 1962 from Barbus tor Hiran river, Katangi.

9) A. singhi Rai, 1962 from Barbus tor in Sihora.

10) A. hirnai Rai, 1962 from Barbus tor near Hirani River, Katangi.

11) A. makundi Gupta, 1963 from Barbus sarana in India.

12) A. heteropneustusis Agarwal, 1964 from Heteropneutus fossili in India.

13) A. ghanensis Fischthal and Thomas, 1972 from host Synodontis sp.

(Monchocidae) in Ghana.

14) A. mirgalai Gupta and Verma, 1976 from Cirrhina mrigala in India.

15) A. saranai Gupta and Verma, 1976 from Barbus sarana in India.

16) A. baranai Gupta and Verma, 1976 from Barilius barna in India.

17) A. gwaliorensis Bhadauaria, 1984 from Mystus vittatus in India.

18) A. tigarai Bhadauaria, 1984 from Channa punctatus in India.

19) A. tosai Takeshi Shimazu, 1988 from Tribolodon hakonensis in Hakkoaido.

20) A. gotoi (Hasegawa et Ozaki, 1926) comb. n. Takeshi Shimazu, 1988 from

Misgurnus anguillicandatus in Nagano.

21) A. (N) lucyae Ernest, 1992 from Notropis zonistius in Illoisis.

22) A. patagonicum Takeshi et al., 2000 from Percichthys colhuapiesis in

Neuquen.

23) A. shianoense sp.Takeshi Shimazu, 2003 from Phoxins lagowski

steindacheri sauvage (cyprinidae) in Japan.

24) A. aburahaya Takeshi Shimazu, 2003 from Phoxins lagowski steindacheri

sauvage (cyprinidae) in Japan.

The present worm under discussion is oral sucker sub-terminal, pharynx

oval, ventral sucker oval, ventral sucker larger than oral sucker, intestinal caeca

bifurcates, posterior extremity, cirrus pouch well developed, ejaculatory duct

tubular, pars prostatica is tube like and zigzag manner. Pars prostatica cell

enclosed by thin walled of cirrus pouch, vesicula seminalis oval and bipartite,

testes circular, ovary is oval, vitellaria confluent or branched at the posterior of the

body. Excretory bladder is tubular.

The present worms closer with the species A. mirgalai Gupta, 1976; A.

saranai Gupta, 1976 and A. baranai Gupta, 1976 which is having oral sucker is

sub terminal, oesophagus absent, pharynx present, ventral sucker larger than oral

sucker, cirrus pouch elongated, ovary is sub spherical, excretory bladder is

tubular but it differ from the species A. mirgalai Gupta, 1976 having testes

(rounded vs elongated), receptaculum seminis flask shaped, vitelline are

(confluent or branched vs follicles); A. saranai Gupta, 1976 which is having

testes (rounded vs elongated, lobed) above mentioned species vesicula seminalis

(oval and bipartite vs S -shaped); A. baranai Gupta, 1976 having which is having

testes (rounded vs oval), vesicula seminalis (oval and bipartite vs coiled) above

species A. saranai Gupta, 1976; A. baranai Gupta, 1976 having receptaculum

seminis pear shaped and vitelline are (confluent or branched vs follicular).

Remaining species differs from the Allocreadium khami n.sp. are as discussed

below,

The present worm differs from the species A. isoporum Looss, 1900 from

the intestine of Tribolodon ezoe in Hakkaido which is having prepharynx very

short, pharynx (oval or elliptical vs barrel shaped), esophagus is long, seminal

receptacle retort shaped, pars prostatica (tube like vs oval), ovary is (oval vs

weakly tribulate).

The present worm differ from the species A. lobatum Wallin E. Ivan, 1909

from the stomach of Semotilus bullaris in the presence of testes (rounded vs

distinctly lobed), cirrus pouch (above the acetabulum vs centre of the acetabulum),

vitellaria (confluent or branched vs large, spherical).

The present worm differs from the species A. nemachilus Kaw, 1950 from

the intestine of Schiozothoran micropogan in Kashmir. Which is having

prepharynx globular, esophagus is long, seminal receptacle pear shaped, testes

(rounded vs tandem), vitelline follicles (confluent or branched vs extend from the

acetabulum upto posterior region of the body), excretory bladder (tubular vs

spherical).

The present worm differs from the species A. thapari Gupta, 1950 from

host, Eutropiichthys vacha in India, in the presence of esophagus is small tubular,

ventral sucker smaller than oral sucker, cirrus pouch (elongated or curved vs oval),

and seminal receptaculum flask shaped.

The present worm differs from the species A. kamlai Gupta, 1956 from host

intestine of Chela bacaila (Ham.) in India, in the presence of prepharynx (absent

vs present), seminal receptacle pear shaped, and vitelline gland (confluent or

branched vs small follicles).

The present worm differs from the species A. mehrai Gupta, 1956 from

host intestine of Rhynochobella oculeata (Bloch.) in India, in the presence of

esophagus short, seminal receptacle cylindrical.

The present worm differs from the species A. ophiocephali Srivastava,

1960 from host intestine of Ophiocephalus punctatus in India, in the presence of

esophagus is short, seminal receptacle pear shaped, excretory bladder (tubular vs

spherical) and vitelline follicles (confluent or branched vs at posterior body extend

from posterior margin of the acetabulum).

The present parasites differ from the species A. makundi Gupta, 1961 from

Barbus sarana in India, in having oral sucker (subterminal vs terminal), Pharynx

is (oval vs nearly spherical), esophagus is present, seminal receptacle pear shaped,

testes (rounded vs spherical, tandem ), vitelline follicles extending from middle of

ventral sucker to hind end of body.

The present worm differs from the species A. dollfusi, Rai, 1962 from

Barbus tor Hiran river, Katangi, in having Prepharynx long, esophagus is present,

intestinal caeca ending blindly behind the posterior testes, testes (rounded vs

oval), vitellaria (confluent or branched vs follicular).

The present parasite differs from the species A. singhi Rai, 1962 from

Barbus tor in Sihora, which is having Prepharynx (absent vs very small),

receptaculum seminis present, testes (rounded vs oval), and vitelline follicles are

extending from anterior level of the ventral sucker to the posterior end of the

body.

The present worms differ from the species A. hirnai Rai, 1962 from Barbus

tor near Hirani River, Katangi, in having oral sucker (subterminal vs terminal),

esophagus is present, receptaculum seminis oval, testes (rounded vs oval),

vitellaria (confluent or branched vs follicular).

The present worms differ from the species A. heteropneustusis Agarwal,

1964 from Heteropneutus fossili in India, in having oral sucker (subterminal vs

terminal), ventral sucker smaller than oral sucker, esophagus is short, testes

(rounded vs oval), vitelline (confluent or branched vs follicular).

The present worm differs from the species A. ghanensis Fischthal and

Thomas, 1972 from host Synodontis sp. (Monchocidae) in Ghana, in the presence

of pharynx (oval vs pyriform), esophagus pouch present, posterior testes (circular

vs longitudinally elongate), Laurer’s canal opposite side of the ovary and Mehli’s

gland, vitelline follicles (confluent or branched vs small), excretory bladder

(tubular vs wider).

The present worm differs from the species A. gwaliorensis Bhadauria, 1984

from Mystus vittatus in India, having oral sucker (subterminal vs terminal),

vesicula seminalis (bipartite vs coiled), ovary rounded, vitelline follicles

(confluent or branched vs commence asymmetrical).

The present parasite differs from the species A. tigarai Bhadauria, 1984

from Channa punctatus in India, which having ventral sucker is (larger vs smaller

than oral sucker), esophagus is short, testes (rounded vs tandem), ovary (oval vs

rounded), uterus short, vitelline follicles (confluent or branched vs commence

asymmetrical).

The present worm differs from the species A. tosai Takeshi Shimazu, 1988

from intestine of Tribolodon hakonensis in Hakkoaido which is having, esophagus

undulating, seminal vesicle (oval and bipartite vs tubular and sinus), pars

prostatica (tube like vs claviform), Laurer’s canal short, vitelline follicles

(confluent or branched vs small), seminal receptacle retort shaped.

The present worm differs from the species A. gotoi (Hasegawa et Ozaki,

1926) comb.n. Takeshi Shimazu, 1988 from Misgurnus anguillicandatus in

Nagano which is having esophagus tube like, testes (circular vs oval and

elliptical), seminal vesicle large tubular and pars prostatica (tube like vs

claviform), testes (circular vs oval), seminal receptacle leaf shaped, Laurer’s canal

short, excretory bladder (tubular vs I shaped).

The present worm differs from the species A. lucyae Ernest, 1992 which is

having esophagus long, testes (rounded vs contiguous, subequal in size),

receptaculum seminis pear shaped, vitellaria commencing at acetabulum and

extending uninterrupted to posterior end of the body.

The present worm differs from the species A. patagonicum Takeshi et al.,

2000 from Percichthys colhuapiesis in Neuquen, which is having eyes spot

pigment solid, esophagus S- shaped, pars prostatica (tube like vs elliptical ), ovary

(oval vs nearly triangular), excretory bladder (tubular vs I shaped).

The present worm differs from the species A. shianoense Takeshi Shimazu,

2003 from intestine of Phoxins lagowski steindacheri sauvage (cyprinidae) in

Japan in the presence of esophagus long, testes (circular vs elliptical), cirrus sac

claviform, Laurer’s canal short, seminal receptacle ovate, vitelline follicles

(confluent or branched vs large distributed from pharynx level of posterior end),

excretory bladder (tubular vs I shaped).

The present worm differs from the species A. aburahaya Takeshi Shimazu,

2003 from intestine of Phoxins lagowski steindacheri sauvage (cyprinidae) in

Japan in the presence of eyes spot pigment dispersed, esophagus undulating, testes

(circular vs irregularly, indented), vesicula seminalis (oval and bipartite vs S-

shaped), pars prostatica (tube like vs globular), seminal receptacle ovate, Laurer’s

canal short, excretory bladder tubular vs I-shaped.

Allocreadium khami n.sp.is proposed after the name of river Kham from

which the host Mastacembelus armatus (Lecepede, 1800) were collected for

recovery of these parasites.

Type species Allocreadium khami n. sp.

Host Mastacembelus armatus (Lecepede, 1800)

Habitat Liver

Locality Kham River, Aurangabad District,

M.S., India

Date of collection December 2008.



Allocreadiidae (Looss, 1902) Stossich, 1903

Orientocreadiinae Yamaguti, 1958

Orientocreadium Tubangui, 1931

Syn. Ganada Chatterji, 1933;

Neoganada Dayal, 1938;

Nizamia Dayal, 1938;

Ganadotrema Dayal, 1949;

Paratormopsolus Dubinina et

Bychowsky in Skrjabin, 1954;

Macrotrema Gupta, 1951.

Orientocreadium striatusae n.sp.

INTRODUCTION:

The genus Orientocreadium was created by Tubangi, (1931) and assigned

to Allocreadiidae Stossich, 1904. Tubangi, (1933) placed the genus in

Allocreadiinae Looss, 1902. After Chatterji, 1933 described the new species

Ganada clariae from the Clarias batrachus in Burma. He stated that the

character of Plagiorchiidae and Plagiorchiinae except for the external seminal

vesicle were evident. The excretory bladder Y-shaped. The genera Ganada,

neoganada, Nizamia, Ganadotrema and Microtrema and all the species

described there in, were assigned by their creators to Plagiochiidae Luhe, 1901

syn. Lepodermatidae Looss, 1901. The subfamily Leptophallinae was created by

Dayal, 1938b for those Plagiorchiidae “having vesicula seminalis divided into

two portion with cirrus sac and a vesicula seminalis externa lying outside the

cirrus sac, free in the parenchyma namely, Leptophallus Luhe, 1909, Ganada,

Neoganada and Nizamia. Dayal, 1949 included his new genus Ganadotrema and

Gupta, 1951b his three new species and species Macrotrema macroni to it.

Jaiswal, 1957 continued to place Neoganada barabankiae Dayal, 1938 in

Leptophallidae characteristics of subfamily is a Y-shaped excretory bladder with

a long stem and short diverticula.

Yamaguti, 1958 is presenting the diagnoses for Orientocreadiinae and

Orienticredium stated in each that the excretory bladder was tubular contrary to

this he included in Orientocreadiinae. The genus Macrotrema and synonymous

with Orientocreadium, the genera Ganada, Neoganada, Nizamia and

Ganadotrema all of which possessed a Y-shaped bladder. All those species

originally described as Orientocreadium to have a tubular to saccular excretory

bladder.

Yamaguti, 1958 review of invalidated Orientocreadiinae, only two valid

Plagiorchicoid genera were represented by namely Orientocreadium Tubangi,

1931 and Ganada Chatterji, 1933; both were emended. These could be placed

either in Plagiorchiidae or partly in Brachycoeliidae, depending on the family

diagnoses by various authors. The former genus was limited to those species with

a tubular to saccular excretory bladder and latter to with Y-shaped. The following

recognized in Orientocreadium and their synynomy indicated such as O.

batracoides Tubangi, 1931 syn O. indicum Pande, 1934; O. raipurensis Saksena,

1958; O. dayalai Saksena, 1958; O. umadasi Saksena, 1960; O. lazeri Khalil,

1961; O. pseudobagri Yamaguti, 1934 syn. Paratormopsolus siluri Bychowsky

and Dabinina, 1954; O. siluri (Bychowsky and Dubunina, 1954) Yamaguti, 1958.

After Yamaguti, 1958; Shimazu, 1990 described new species O. chaenogobii from

the rectum of chaenogobius laevis in Shibecha, Kushiro.

Later Valadimir et al., 2009 described the two species, O. pseudobergi

Yamaguti from the rectum of Perccottus glegni and O. elegans Valadimir et al.,

2009 from yellow catfish, Pelteobargus fulvidraco in Russia.


Orientocreadium striatusae n.sp. from Channa striatus (Bloch, 1793).

DESCRIPTION:

Seven worms were recovered from the buccopharyngeal area of Channa

striatus (Bloch, 1793) from Paithan, Aurangabad district, M.S., India, in the

month of December 2009. Trematode were collected from the buccopharyngeal

cavity of fish, washed in distilled water, flattened between coverglass and slides,

fixed in 4% formalin until 24 hours, trematode worms after removing from

preservatives, washed in distilled water, stained with Harri’s Haematoxylene,


cleared in xylene, mounted in DPX. Drawings were made with the help of

Camera Lucida and all the measurement taken are in millimeter.

The present worm is elongated, spinous, whitish in colour, in live

condition, they are leech like movement i.e. contraction and expansion. The

anterior-posterior ends are rounded. It measures 11.2272 mm. in length and

2.9595 mm. in breadth. The oral sucker oval, sub-terminal and it measures 0.3977

mm. in length and 0.4204 mm. in breadth. Pharynx is oval and overlaps to the oral

sucker, measures 0.1818 mm. in length and 0.1590 mm. in breadth, esophagus

pouch is somewhat oval, overlap to the pharynx and it measures 0.3295 mm. in

diameter. Ventral sucker rounded and located 1\3 part of the worm below the

cirrus pouch and it measures 4.2044 mm. in diameter. Ventral sucker larger than

oral sucker. Intestinal caeca elongated, bifurcate and posterior extremity.

The male reproductive system, cirrus pouch is well developed at near the

gonopore and between the esophagus pouch and acetabulum. Gonopore small,

rounded, cirrus is located anterior to the genital pore, and it is tube like and

measures 0.1022 mm. in length and 0.02272 mm. in breadth. Cirrus pouch large

and measures 1.0680 mm. in diameter. Ejaculatory duct short, tube like anterior of

pars prostatica or attach to the genital pore, measures 0.1250 mm. in length and

0.02272 mm. in breadth. Pars prostatica oval and not distinct from the ejaculatory

duct and surrounded by the pars prostatica cells. Vesicula seminalis is oval and

bipartite measures 0.7386 mm. in diameter, internal seminal vesicle surrounded by

the pars prostatica cells and external seminal vesicle free from the parenchyma.

Testes are two, large located below ovary and uterus. Anterior testes

transversely elongated, overlap the uterus and measures 1.1703 mm. in length and

0.6250 mm. in breadth. Posterior testes bean shaped, slightly lobed and it


The female reproductive system, ovary is longitudinally elongated or oval

located above anterior testes it measures 1.1817 mm. in length and 0.9207 mm. in

breadth. Seminal receptacles is well developed beside the ovary and overlap the

uterus, it measures 1.0680 mm. in diameter. Mehli’s gland is rounded and located

below the seminal receptacles. Uterus much coiled at between the ovary and

anterior testes and overlap to the vitellaria. Vitellaria are complex, confluent

extend from the ventral sucker to posterior region. Eggs are not developed in the

uterus (unembryonated).

Excretory pore small, rounded at the posterior extremity. Excretory bladder

Y-shaped.

DISCUSSION:

Tubangi created the genus Orientocreadium in 1931 with its type of species

Orientocreadium batracoides from Clarias batrachus in India.

1) Orientocreadium batracoides Tubangi, 1931 from Clarias batrachus in

Philippines.

2) O. indicum, 1934 by Pande from the Rita buchanani in India syn. of O.

batracoides Tubangi, 1931.

3) Orientocreadium pseudobagri Yamaguti, 1934 from the host Pseudobagrus

aurantiaus in Japan.

4) Orientocreadium raipurensis Saksena, 1958 from Clarias batrachus in

India syn. of O. batracoides Tubangi, 1931.

5) Orientocreadium dayalai Saksena, 1958 from Clarias magur in India syn.

of O. batracoides Tubangi, 1931.

6) Orientocreadium umadasi Saksena, 1960 from Clarias magur in India syn.

of O. batracoides Tubangi, 1931.

7) O. lazeri Khalil, 1961 from the host Clarias batrachus in Sudan.

8) O. chaenogobii Shimazu, 1990 from the rectum of chaenogobius laevis in

Shibecha, Kushiro.

9) O. Peudobergi Yamaguti Valadimir, 2009 from the rectum of Perccottus

glegni in Russia.

10) O. elegans Valadimir, 2009 from the yellow catfish in Russia.

The present worm under discussion having oral sucker oval, sub terminal,

pharynx oval, esophagus pouch oval and overlap to the one another, ventral sucker

rounded, cirrus pouch large, preacetabular, ejaculatory duct short, pars proststica

oval, vesicula seminalis oval and bipartite, ovary longitudinally elongated, testes

are two, large and anterior testes transversely elongated, posterior testes bean

shaped, slightly lobed. Vitellaria are complex, confluent from acetabulum to hind

body, excretory bladder Y-shaped.

The present worm closer with the species Orientocreadiun batracoides

Tubangi, 1931; O. indicum, 1943 by Pande; Orientocreadium pseudobagri

Yamaguti, 1934 ; Orientocreadium raipurensis Saksena, 1958 ; Orientocreadium

dayalai Saksena, 1958; Orientocreadium umadasi Saksena, 1960; O. lazeri Khalil,

1961 in having oral sucker oval, sub-terminal, esophagus present, pharynx oval,

ventral sucker circular, seminal vesicles is bipartite, however, it differs from above

species the ovary is (longitudinally elongated vs rounded), testes (two, elongated

to bean shaped vs slightly oblique), and excretory bladder (Y shaped vs tubular).

The present worm resembles with the species O. Pseudobergi Yamaguti

Valadimir, 2009; O. elegans Valadimir, 2009 in having oral sucker oval, sub-

terminal, esophagus present, ventral sucker circular, ovary is oval, seminal

vesicles bipartite, excretory bladder Y-shaped, but it differs from the pharynx is

(oval vs cruciform identification), testes are (elongated, bean shaped to slightly

lobed vs oval) and vitellaria (confluent or complex vs follicular). The present

worm differs from the species O. chaenogobii Shimazu, 1990 which is having

seminal vesicles (bipartite vs tubular), uterus is much coiled at the level of

acetabulum to posterior end, and vitellaria are (confluent in middle portion of the

body vs oval and lateral side of the intestinal ceca).

Above distinct character noted, justify the recognition of the present worm

as a new species and hence the name Orientocreadium striatusae n.sp.is proposed

the name of host species, Channa striatus (1793).

Type species Orientocreadium striatusae n.sp.


Habitat Buccopharyngeal area

Locality Paithan, Aurangabad district,

M.S., India.

Date of collection December 2009.

CHAPTER- II

HISTOCHEMISTRY

PHOSPHATASES

INTRODUCTION:

Enzymes are catalyst of biological origin which accelerate the various

cellular reactions, without themselves undergoing any apparent change during

the course of action and that are not dependant on the intact cell for this activity.

They are functional at ordinary body temperature and are active usually within

cells often they are secreted by the cells and are active outside.

In 1878, Kuhne used the word enzyme to indicate the catalysis taking

place in the biological systems. In human body uses thousand of enzymes to

carryout a myriad of biochemical process. E.g. enzyme assisted process is

digestion. Enzyme help breakdown carbohydrates, fat and protein into simple

compound that the body can absorb and turn for energy or use to built or repair

tissue. A shortage of enzyme in the body will influence the health of the entire

body and symptoms may include stomach gas, indigestion, bloating, heartburn

and flatulence.

Phosphatases are the hydrolytic enzymes, and are responsible for the

breakdown of phosphate esters, can be divided into three type, mono, di and

triphosphatases. Phosphatase plays an important role, regulating metabolic

processes with alkaline phosphatase taking part in active transport through

cellular membranes and acid phosphatase (isosome marker) indirectly providing

information regarding intercell digestion process. The phosphomonoesterases are

divided on the basis of their optimum pH level into invitro (cf Kroon et al.,

1944), those which works on the alkaline side of neutrally and particularly at pH

9.0 and pH 5.0 constitute the acid phosphatases. The phosphatases are

nevertheless greatly improved if one assumes that high activity indicates

increased phosphate transfer rather than hydrolysis of phosphate esters.

Read, (1966) however, argues strongly against this hypothesis and has

suggested that, the phosphatases serve to prevent the uptake of phosphorylated

compound, an excess of which would upset the metabolism. A more probable

function for the phosphatases in the hydrolysis separately by the tubule cells

(Dike and Read, 1971). In many parasites, the presence of phosphatase is

regarded as the indicator of the areas responsible for secretion and excretion

activities and the absorption of nutritive components. Alkaline phosphatase is an

enzyme found throughout the body. Like all enzymes, it is needed is small

amount to trigger specific reactions.

The presence of phosphatases has also been noted in Taenia pisiformis by

Pennat-de Cooman and van Grembergen, (1947) and Erasmus, (1957a);

Moniezia expansa by Rogers, (1947); Erasmus, (1957b); Echinococcus

granulossus by Kelejian et al., (1961); Hydatigara taeniaeformis by Waitz,

(1963); Waitz and Schardein, (1964); Hymenolepis microatoma by Bogitch,

(1963); and in hymenolepis nana, H. diminita and Diphyllobothriidium canium

by Waitz and Schardein, (1964). The distribution of phosphatases in the cuticle

of the pseudophyllidean Ligula is in agreement with the result obtained by many

other workers with cyclophyllidean cestodes. However, although both alkaline

and acid phosphatases are present in the cuticle of Ligula.

Threadgold, 1968 has suggested that the phosphatases release free

phosphate that can be coupled to sugar by another enzyme before or during

absorption. Thus the phosphorylated transfer of substances into the cell from

lumen of the duct may occur. The occurrence of vital enzymes, viz, acid

phosphatase (Acpase) and alkaline phosphatase (Alkpase) has been resolutely

demonstrated in a number of helminth parasite, both histochemically and

biochemically (O. Poljakova-Krustena et al., 1983, M. Stettler et al., 2001),

including Railleitina echinobothrium (P. Pal and V. Tandon, 1998). These

enzymes have been unequivocally revealed to be intimately associated with the

tegument and subtegumental regions of cestode and trematodes as well as the

cuticle of nematodes (K. H. Kwak & C.H.Kim, 1997; K. Buchmann, 1998; R. H.

Fetterer et al., 2000).

The exact physiological functions of alkaline phosphatase are uncertain it

has been suggested that the enzyme is linked in someway with the movement of

hexoses, as for example across the body wall of a cestode Taenia pisiformis

(Erasmus, 1957) or with the movement of ions and water, e.g. in the mucosa of

the eels (Oide, 1970). Certainly the enzyme is a hydrolytic one, and this is

emphasized in the demonstration by (Arme and Read, 1970) that there exists in

the cestode Hymenolepis diminuta a surface tegumentory alkaline phosphatase

which readily hydrolyses impermeable fructose diphosphate in the external

media. Among majority of nematodes the activity of acid phosphatase in the

cuticle is high and is correlated with the absorption of glucose through the body

walls (Maki and Yauagisawa, 1980).

Gupta and Gupta, (1977) who reported higher activity of alkaline

phosphatase than acid phosphatase in intestinal trematode, Ganeo tigrinum only

one peak for acid and alkaline phosphatase activity was noted in P. egretti, which

with most of the previous finding (Srivastava and Gupta, 1975; Gupta and

Agarwal, 1979). The reason for this change in the relative proportion of these

enzymes in different phases of the life history is not clear but it may be

associated with the growth and development of reproductive system.

The present study deals with the investigation on the Senga rupchandensis

n. sp., Circumoncobothrium jadhavae n. sp., Genarchopsis paithanensis n.sp.,

Allocreadium khami n sp., Orientocreadium striatusae n. sp. collected from

freshwater fishes namely and Channa striatus (Bloch, 1793) and Mastacembelus


ALKALINE PHOSPHATASE

MATERIAL AND METHODS:

The freshwater fishes namely, Mastacembelus armatus (Lecepede, 1800)

and Channa striatus (Bloch, 1793) were collected from different place of

Marathwada region of Maharashtra state. Fishes were brought to the laboratory

and dissected or cut , opened and observed the intestine , liver, stomach, gill

cavity, heart, spleen etc. examined carefully and collected the parasite kept in

saline water, washed well in distilled water, the identical worms were separated

and processed for taxonomical study. The cestode parasite were collected from

liver, intestine and trematode parasite were collected from liver, intestine,

stomach and gills etc. The use of Gomori, (1946) Cobalt nitrate method for

alkaline phosphatase, the cestode parasite preserved in chilled Acetone for a

short time before fixing 2-4 mm. thick cestode tissue slices and whole trematode

parasite were fixed in chilled acetone for 24-48 hours with 2-3 changes. Then

passed through Chloroform for 1 hours, embedded in paraffin wax having a

melting point not higher than 56 OC and blocks were prepared.

Section were cut at 6-8 µ, deparaffinized in xylene, hydrate to water

(100%, 80%, 60%, 40%, 20%) alcohol, rinsing with several dips in water, then

transferred or placed in incubating mixture such as, 10 ml of 3% Sodium β-

glycerophosphate, 10 ml of 2% Sodium diethyl barbiturate, 20ml of 2% Calcium

chloride, 05 ml of Distilled water, keep at 37 OC. for 30-60 minutes. This stock

solution ready mix to make incubation mixture at the time of use, wash well in

distilled water, treat with 2% aqueous Cobalt nitrate for 3-5 minutes, rinsed in

distilled water and transferred to 2% aqueous Yellow Ammonium sulphide for 1-

2 minutes, washed in distilled water, dehydrated in (20%, 40%, 60%, 80%,

100%) alcohol grades, cleared in xylene and mount in glycerin jelly and

observed under Microscope which shows the alkaline phosphatase activity in

various structure which are stained black or brownish in colour.

RESULT AND DISCUSSIONS:


Taxonomic study suggested that, the worm is to be a new species of genus

Senga as Senga rupchandensis n. sp.

The observation of the longitudinal section of the Senga rupchandensis

n.sp. was indicated and it is found that, in the mature proglottids, the intense

alkaline phosphatase enzyme activity in ovary, cirrus pouch, uterus, isthmus,

testes, vitellaria and very less in the musculature, cuticular parenchyma has high

activity (Plate-10 A).

The observation of the longitudinal section of the gravid proglottids was

observed and it is found that, the intense alkaline phosphatase enzyme activity in

the vitellaria, body wall, egg shell, testes, and absent in musculature (Plate-10B).

From above observation of the longitudinal section of the Senga

rupchandensis n.sp. it can concluded that in mature and gravid proglottids shows

intense alkaline phosphatase activity in reproductive organ.


The observation of the longitudinal section of Circumoncobothrium

jadhavae n.sp. was indicated that, in mature proglottids the intense alkaline

phosphatase enzyme activity in the ovary, testes, vitellaria and moderate in the

cirrus sac, uterus; no activity in the musculature (Plate-11A).

The observation of the longitudinal section of the gravid proglottids was

indicated; it is found that, the intense alkaline phosphatase enzyme activity in

vitellaria and moderate activity in the egg shell (Plate-11B).

From the observation of the above species, it is concluded that the intense

alkaline phosphatase enzyme activity in the mature proglottids.


The observation of longitudinal section of the whole parasite and anterior

region of Genarchopsis paithanensis n.sp. was indicated and it is found that, the

moderate alkaline phosphatase enzyme activity in oral sucker; no alkaline

phosphatase activity in cirrus pouch (Plate-12 A&B); but in middle portion, the

intense enzyme activity in ventral sucker (Plate-12 C).

The observation of the longitudinal section of posterior region, it is found

that, the intense alkaline phosphatase activity in the uterus, egg shell, Mehli’s

gland, and moderate reaction in ovary, testes (Plate-12 D).

It is concluded that the observation of the above species maximum

alkaline activity in reproductive organ.


The observation of longitudinal section of the anterior region of

Allocreadium khami n.sp. was indicated and it is found that, no alkaline

phosphatase activity in oral sucker and cirrus pouch (Plate-13 A).

The longitudinal section of the middle portion of the species, it is

observed that, the intense activity in uterus; moderate alkaline phosphatase

activity in the intestinal caeca; weak enzyme activity in the ovary and no reaction

in ventral sucker (Plate-13 B).

The observation of longitudinal section of the posterior region of the

species is found that, the intense alkaline phosphatase activity in uterus, testes,

and vitellaria (Plate-13 C).


The observation of the longitudinal section of the whole parasite and

anterior region of the Orientocreadium striatusae n.sp. was indicated and it is

found that, the intense alkaline phosphatase enzyme activity in the vitellaria,

sucker, and no reaction in musculature (Plate-14 A&B).

The observation of the longitudinal section of the posterior region of the

Orientocreadium striatusae n.sp., it is found that, the intense alkaline

phosphatase activity in the vitellaria, uterus; weak reaction in the ovary, seminal

receptacles, caeca and moderate in the testes (Plate-14 C)

From above observation of the species Orientocreadium striatusae n.sp.

the maximum alkaline phosphatase activity in the reproductive organ in

reproductive phase.

ACID PHOSPHATASE


The freshwater fishes namely, Mastacembelus armatus (Lecepede, 1800)

and Channa striatus (Bloch, 1793) were collected from different place of

Marathwada region of Maharashtra state. Fishes were brought to the laboratory

and dissected or cut , opened and observed the intestine , liver, stomach, gill

cavity, heart, spleen etc. examined carefully and collected the parasite kept in

saline water, washed well in distilled water, the identical worms were separated

and processed for taxonomical study. The cestode parasite were collected from

liver, intestine and trematode parasite were collected from liver, intestine,

stomach and gills etc. The use of Gomori,s (1950) Lead nitrate method for acid

phosphatase, the cestode parasite preserved in chilled Acetone for a short time

before fixing 2-4 mm. thick cestode tissue slices and whole trematode parasite

were fixed in chilled acetone for 24-48 hours with 2-3 changes. Then passed to

Chloroform for 1 hours, embedded in paraffin wax having a melting point not

higher than 56 OC and blocks were prepared.

Section were cut at 6-8 µ, deparaffinized in xylene, hydrate to water

(100%, 80%, 60%, 40%, 20%) alcoholic grades, then transferred or placed in

incubating mixture such as, 30 ml of 0.01M Sodium β- glycerophosphate, 30 ml

of 0.05 M Acetate buffer (pH 5.0), 10ml of 0.004 M Lead nitrate. Lead nitrate

was dissolved in the buffer β- glycerophosphate was added. The final PH of the

incubation was maintained at 5.0 at 37 OC. for 30 minutes, washed well in

distilled water and immersed in 25 aqueous Yellow Ammonium sulphide for 2

minutes, rinsed in distilled water, dehydrated in (20%, 40%, 60%, 80%, 100%)

alcohol grades, cleared in xylene and mount in glycerin jelly and observed under

Microscope which shows the acid phosphatase activity is indicated black colour

in the section.

RESULT AND DISCUSSION:


The observation of the longitudinal section of the Senga rupchandensis

n.sp. was indicated and it is found that, in mature proglottids, the moderate acid

phosphatase enzyme activity in the vitellaria, ovary, isthmus and weak acid

phosphatases activity in the testes; no reaction in cirrus pouch and uterus (Plate-

15 A&B).

The observation of the longitudinal section of the gravid proglottides and

it is found that, the intense activity in the vitellaria, egg shell (Plate-15 C).

It is concluded that the Senga rupchandensis n.sp., the observation of the

parasite the maximum activity in the reproductive organ and no activity in the

musculature.


The observation of the longitudinal section of Circumoncobothrium

jadhavae n.sp., it is found that, in mature proglottides the intense acid

phosphatase enzyme activity in the ovary, vitellaria; moderate activity in testes,

uterus and no reaction in the cirrus, cirrus pouch (Plate-16 A).

The observation of the longitudinal section of the gravid proglottids, the

intense acid phosphatase enzyme activity in egg shell and vitellaria; moderate

acid phosphatase enzyme activity in the testes and less in musculature (Plate-16

B).

It is concluded that, Circumoncobothrium jadhavae n.sp., the observation

of the parasite, the maximum activity in reproductive organ except cirrus and

cirrus pouch.

Genarchopsis paithanenesis n.sp.

The observation of the longitudinal section of the trematode Genarchopsis

paithanensis n.sp. was indicated and it is found that, the whole parasite and

anterior region of the Genarchopsis paithanensis n.sp. intense acid phosphatase

enzyme activity in the suckers; no reaction in the cirrus pouch and musculature

(Plate-17 A&B).

The observation of the longitudinal section of the posterior region, the

moderate acid phosphatase enzyme activity in the uterus, Mehli’s gland and egg

shell; weak activity in the ovary and testes (Plate-17 C).

It is concluded that, the observation of the above species moderate acid

phosphatase enzyme activity in reproductive organ.


The observation of longitudinal section of the anterior region of

Allocreadium khami n.sp. was indicated and it is found that, no acid phosphatase

enzyme activity in oral sucker and cirrus pouch (Plate-18 A).

The longitudinal section of the middle portion of the species, it is

observed, the moderate acid phosphatase enzyme activity in the intestinal caeca,

uterus; weak enzyme activity in the ovary and no reaction in ventral sucker

(Plate-18 B).

The observation of longitudinal section of the posterior region of the

species it is found that, the intense activity in testes; moderate acid phosphatase

activity in uterus, small activity in vitellarian duct and vitellaria (Plate-18 C).


The observation of the longitudinal section of the whole and anterior

region of the Orientocreadium striatusae n.sp. was indicated and it is found that

the maximum acid phosphatase enzyme activity in the vitellaria; small activity in

the sucker and no reaction in the musculature (Plate-19 A&B).

The observation of the longitudinal section of the posterior region, it is

found that, the intense acid phosphatase activity in the vitellaria, uterus, moderate

activity in the ovary, testes and weak activity in Mehli’s gland (Plate-19 C).

It is concluded that the maximum acid phosphatase enzyme activity in the

reproductive organ.

The present studies showing the distribution of the acid and alkaline

phosphatase enzyme activity in the above species. This results are similar and

discussed with Erasmus, (1957) pointed out that the maximum alkaline

phosphatase activity in Moniezia expansa occurred in the region of mature

proglottides. C. Arme, (1966) the histochemical studies on pseudophyllidean

Ligula intestinalis from Ratilus ratilus and observed the alkaline phosphatase

activity is slight in most part of the reproductive system with strong in cirrus and

egg shell, a weak positive reaction in musculature and acid phosphatase is

restricted almost entirely to the cuticle and sub cuticle but weak reaction in

reproductive organ. Kelejian et al., (1961) found no acid phosphatase in the

cuticle of Echinococcus granulossus.

Mayberry and Tibbitts, (1972) also found an increased intensity of AlPase

in mature and gravid proglottides than in immature ones of Hymenolepis

diminuta. Roy T.K., (1979) observed the presence of acid phosphatase activity in

almost all the tissues of the parasites, the very intense enzyme in the excretory

canal, male and female reproductive organ. Alkaline phosphatase activity in the

excretory canal, male and female reproductive organ, absence in the parenchyma

and the enzyme activity in the tegument of immature proglottides are less intense

than that of mature proglottides.

In contrast from Yamao, (1952) reported that no alkaline phosphatase

activity could be demonstrated in the cuticle of several species cyclophyllidean

cestodes. Araxie et al., (1961) observed the alkaline phosphatase activity in

cuticle, sub cuticle cells, and slight activity in nuclei of mature ova and positive

reaction in sucker and acid phosphatase activity did not shows in the cestode

Echinicoccus granulossus.

Christina Ohman-James, (1968), histochemical studies of the cestode

Diphyllobothrium dentriticum from white fish (Coregonus lavaretus) reported

alkaline phosphatase activity only in nerves and in the anterior end of

Diphyllobothrium dendruticum to an increased host-parasite contact in that part

of the parasite. AlPase distribution in Railleitina (R.) johri appears to be more

concentrated posteriorly in the parasite. It is quite possible that the different

parasites may show different pattern of enzyme distribution, or the host may also

influence the distribution of an enzyme in the parasite.

The present result of trematodes similar with acid phosphatase activity in

Fasciola buski, was mainly in the sub tegument these findings agree closely

those reported by other workers in sterigeids, Fasciola hepatica and Schistosoma

mansoni (Erasmus, 1972). High concentrations of phosphatases were

demonstrated in the egg and uterine wall of the species studied and the enzymes

were more densely concentrated in the shell of the mature eggs. The vitellaria

and reproductive system, including the gonads were also shown to have very

strong reactions for both acid and alkaline phosphatases. These findings are

similar to those reported in Fasciola hepatica, Schistosoma mansoni and many

other trematodes (Becejac and Krvavica, 1964; Probert and Lwin, 1974; Barry

and Mawdesley-Thomas, 1968; Sharma, 1976; Tandon and Misra, 1978) Fujino

et al., (1989) were reported alkaline phosphatase activity in the excretory bladder

and acid phosphatase activity in the intestinal caeca of the Paragonicus

westernmani trematode. Trematodes investigated contain strong acid and alkaline

phosphatase activity in sucker. In G. explanatum, the reaction products are

localized all along the tegumental surfaces of the suckers which are in direct

contact with the host tissue and may have glandular activity. The distribution is

more or less similar to that reported in adhesive organ of strigeids (Erasmus and

Ohman, 1963; Ohman, 1966) in which it appears that the phosphatases present in

the sucker may be involved in carbohydrate metabolism and the absorption of

nutrients, as well as in dissolved host tissue at the host-parasite interface for

extracorporeal digestion. This probably occurs also in Fasciola hepatica

(Thorsell and Bjorman, 1965).

In contrast, Sharma and Hanna, (1988) observed the acid and alkaline

phosphatase activity in the tegument of Orthocoelium scoliocoelium and

Paramphistimum cervi trematode.

CONCLUSION:

It is concluded that, the high alkaline and acid phosphatase activity

observed in reproductive organ such as, testes, ovaries, cirrus sac, egg shell and

vitellaria of adult parasite indicates that permeation of metabolites across the

membrane of almost all organ of the system takes place in order to allow

synthesis of protein required for the formation of proteinaeous structure. ATPase

activity in vitelline cells takes part in the active synthesis of the material as well

as in the production of the new cells. Phosphatase in the excretory canal may also

helps in the distribution of nutrients from one part of the worm to another. In

uterus, egg shell and may be involved in transport mechanism.

CHAPTER-III HISTOPATHOLOGY

HISTOPATHOLOGY

INTRODUCTION:

The study of contact, interaction, beneficial or harmful and relationship

between the host and the parasite is called as histopathology. Some biologist say

that all animals are parasites because they have to rely an other living organisms

for their food, be these plant and animals. A parasite is physiologically dependant

on its host and cannot survive in its absence. The physiological compatibility is

sometimes such that of the host and its parasite may evolve though not

necessarily at the same place. Many species of parasite can survive as adult in

warm blooded animals and in their larval stages either in cold blooded animals or

may be in water and soil.

The host parasite association as it grows older generation after

generations, the impact of pathogenicity would theoretically become less and less

due to mutual characteristics and biochemical adaptability. The structural and

biological characteristics of the parasites, the various modifications in the organs

of the attachment. The mechanism of feeding, their life cycle inside and outside

the host. The biochemical adaptation of the parasite enabling them to stay alive

inside the host. The mechanism of entry and the establishment of a parasite

within a particular fishes. The degree of the response by each host to the parasite

while making contact is related to the nature of the tissue site of invasion and

also to the intimacy of the host or parasite contact. It is also related to the stage of

development of the living organisms, whether it is an adult or larval form.

Some parasites are ectoparasite and endoparasite. Ectoparasite living on

the surface of the body of their host, such as blood sucking or feeding upon hair,

feather skin or secretion of the skin. Endoparasite, living inside the body,

occupies digestive tract or other cavities of the body or lives in various organs,

blood, tissue, so even within cells. Some parasites pass different phases of their

life cycle in two or more hosts. Parasites have little need for sense organ and

seldom have they as highly developed as do related free-living animals.

The metabolic type of the parasite depends on number of factors,

including the presence or absence of oxygen in the habitat, on the parasite ability

to bind it with its respiratory pigment, on its type of feeding and on its size.

Helminth means ‘worm’. Helminthes includes the animals belonging to

the phylum platyhelminthes and nematode of phylum aschelminthes. Many of

the parasite form of this group are popularly known as parasite worms.

Platyhelminthes is a large and diverse group of organisms, some of which are

free-living but most of which are parasite, living on, or in most species of

vertebrates and invertebrates. They are flattened dorsoventrally.

Various taxonomical studies revealed that the holdfast organ are

ingeniously adopted and this adaptation helps them to attach to the mucosa of the

specific host, where as there are other species which are having weakly

developed scolex and hence they have a wide range of host specificity.

Cestodes absorb semi digested food material from the intestine and it has

long been assumed that cestode lie in a bath of semi digested ‘soup’ from which

they can extract nutrients metabolites and invitro. Studies suggest that a complex

nutritional relationship occurs between a cestode and its host. The physiological

conditions of a specific species depend mainly on the type of sites which

available, this may be favourable or unfavourable, whereas the parasite get

sufficient nourishment.

Metacestode of tapeworms of the cyclophyllidean family

Gryporhynchidae occur frequently in the gut, gall bladder or the abdominal

cavity of fish. These cestode stages frequently referred to as ‘cysticerus’ may be

more common in host fishes. The histological finding obtained the study show

that Neogryporchynthus cheilancristrotus is a truly pathogenic species causing

degeneration and inflammation intestinal wall, while the majority of cestode

inhabiting the gut of fish live in the lumen of the gut and attached more or less

firmly to the gut epithelium with scolices, Neogryporchynthus cheilancristrotus

intrudes into deeper layer of the gut wall and breaking through the epithelium is

located in the lamina propria of the mucosa layer (Molnar, 2005).

Histopathological investigation such as marked lesions in the mucosa,

inflammation and congestion of mucous glands and intestinal hemorrhages lead

to severe anemic condition in the fish (Williams, 1967; Roberts, 2001). Besides

mechanical injuries, atrophy of tissues and lesions of the alimentary canal, blood

vessels or duct etc., the parasite also induce toxic metabolic by product

(endotoxic / ectotoxic) eliciting changes in blood, enzymes, vitamins and

hormonal activity of the host (Symons and Fairbain, 1963; Poynter, 1966;

Mongkol-Primpol, 2000; Ekman & Norrgren, 2003).

The extensive study on the host parasite relationship has been carried out

by Amoebotaenia indiana by Mitra and Shinde, (1980); Hymenolepis nana by

Bailey, (1951). Host response to implanted adult H. nana was studied by

Coleman, DE. Sa (1964) and experimental immunization of dog against E.

granulossus was first observed by Foresek and Rukavina in (1959). Murlidhar

and Shinde, (1987) observed histopathology of Acanthobothrium uncinathum

from a fish Rhyncobatus ajeddensis. Baur et al., (1959); Amlacher, (1961);

Hayunga, E.G. (1977) and Mackiewicz, (1972) have studied the histopathology

of intestine of fish caused due to cestodes. Gopal Krishnan, (1968); Satpute, A.

and Agarwal (1974); Bose and Sinha, (1981), Niyogi, A. and Agarwal, S. M.

(1989) have studied the caryophyllaeidiasis in fish host.

Smyth, (1969) and Ress, (1967) stated that the response to the adult

cestodes only develop of the mucosa of the hosts intestine invaded.

Helminthes parasite poses a serious threat to the fish population. The

study of trematode parasite found in fish is a field wide open for investigations

always. Although many studies have been made on the trematode in fish,

relatively few have been concerned with the histophysiological, histopathological

and more particularly the changes in the chemical composition of the host tissues

(Bose and Sinha, 1979; Barbara, 1980; Muzzal, 1980; Chung Yui-tan, 1981;

Christina, 1982; Paperna and Vanus, 1983; Maqbool and Nizami, 1984; Zarina,

1990; and Bhargavi and Krishna, 1993). The parasite may effect host interactions

are deleterious to the host (Holmes and Bethel, 1972; Holmes, 1979; Minchella

and Scott, 1991). The festation of parasite to the host cause many alterations

including the macromolecular compositions of host’s tissues, which is the

evidence that was supported by histochemical and biochemical studies.

Mostly digenetic trematodes produce infection in man and animal of the

various fluke infections. The minute trematodes attach to the cell in the mucosal

crypts, usually at the duodenal and jejunal walls of the small intestine, usually

excess secretion of mucus. The damage produced by trematode is mechanical

obstructive and toxic. At each site of attachment a mucosal ulcer is produced. In

the study of histopathological and histochemical changes that are caused to the

intestine of fish Clarias batrachus due to infestation of trematode parasite,

Genarchopsis goppo. Studies have been made to visualize the biochemical

changes occurred in fish due to parasitic infestation (Robinson and Williams,

1971 and Gupta and Agarwal, 1984), but relatively a very less information is

available on the studies made histochemically, however these studies especially

elucidate the changes occurred in the infected tissues of the host due to the

parasite.

The pathological consequences of parasite effects on fishes are well

documented and parasitic fauna certainly influences the fish health, causes to the

mortality and thus reduce fish population. It is well known fact that the helminth

parasite bring pathological changes brought by the parasite in the host may be

due to the mechanical damage or due to the release of toxins by the parasite.

Gupta and Agarwal, (1984) and Singh et al., (1984) who have also

described the pathological condition brought by trematodes in various fishes.

Procamallanus is a common nematode parasite in the stomach and

intestine of fishes inhabiting freshwater, brackish water and marine water

ecosystem of the world (Sinha, 1988; Zaman and Leong, 1988; Chandra, 1994;

Mortens and Moens, 1995; Bijukumar, 1996; Gozalez-Solis et al, 2002). Though

the intermediate host (vector) of the parasite copepods (Chandra and Modak,

1995; Sinha, 2000), there exist reports that adult as well as larval stages of

Procamallanus are pathogenic to fish (De and Maity, 2000; Ruhela et al., 2006).

Histologically larval nematode migration and encapsulation within body

tissues and visceral organ often cause the development of lesions described as

fibro-granulomatus, although in many instances no host inflammatory reaction is

elicited (Fergusen, 1989). Histopathology of infection of Pomphorynchus

kashmirensis have been the subject of earlier work at the laboratory (Chishti &

Bakshi, 1992; Chishti et al., 2002; Chishti et al., 2003) which provide a

convenient fish- helminthes model in which the possible role of acquired

immune response was investigated. Voltoneu et al., (1994) described

pathological changes due to Raphidasaris acus in liver of roach. Murai et al.,

(1997) described presence of Paradilepis scolecina larvae in the series

membrane of the intestine and gall bladder in beam caught in Lake Balaton.

Stephenson et al., (1980) remarked that pathology of the intestinal layer might be

due to combination of physical, biochemical and histopathological responses.

The pathology of P. laevis infection in the alimentary tract of the salmon (salmo

salar), the club (Leuciscus cephalus), the rainbow trout (Salmo gairderi) and the

stone loach (Nemacheileus barbatulus) was demonstrated in earlier studies by

Dezfuli B.S., 1991; Hine, P.M. and Kennedy, C.R. 1974; Pipy, J.H.C. 1969;

Wanstali S.T. et al, 1986).


n. sp., Circumoncobothrium jadhavae n.sp., Genarchopsis paithanensis n.sp.,


freshwater fishes namely Channa striatus (Bloch, 1793) and Mastacembelus



For the histopathological study, the freshwater fishes, namely

Mastacembelus armatus (Lecepede, 1800) and Channa striatus (Bloch, 1793)

were collected from the different region of Marathwada (M.S.), India, and

brought to the laboratory and killed by pithing brain they were examination

externally ( i.e. scale, gills, fins) and later cut opened the fish and observed

internally taken out the intestine ,liver, heart, spleen in the normal saline water in

petridish and cut opened; examination carefully for parasites. The cestode were

collected from the intestine, liver and trematode worms were collected from

intestine, stomach, liver, gill cavity etc. identified worm were kept separately and

wash in saline water solution, flattened by using coverglass and slide then

preserved in 4 % formalin for taxonomical studies. The slides were prepared by

Harri’s Haematoxylene stain, dehydrated in alcoholic grades (30%, 50%, 70%,

90%, and 100%), cleared in Xylene and mounted in DPX. Drawing were made

with the aid of Camera Lucida, all measurement are taken in millimeter,

identification was carries out by the using Systema Helminthum cestode Vol-I

(1956), and trematode Vol- I and II (1971) by Yamaguti.

The infected intestine, liver attached with the cestode parasite and infected

liver, stomach, intestine, gills with attached trematode parasite, were kept intact

and small pieces of such intestines, stomach, liver, gills were fixed in Bouins

fluid for histopathological studied. The fixed tissues were washed in distilled

water, dehydrated in alcoholic grades, cleared in xylene, embedded in paraffin

wax with melting point (58-60 OC).

Block were cut at 8mµ and slides were stained with Haematoxylene

counter stained with eosin stain. Best slides were selected, observed under the

microscope and photographs are taken.


The histopathological study is carried out with microtome technique,

where as the section were cut 8mµ on a rotary microtome and stained with

Harri’s Haematoxylene counter stained with Eosin. The best select slides of

healthy and infected intestine, gills and liver was observed in the microscope and

photograph of the best slides were taken and are considered for the discussion.


The taxonomical observations reveals that, the present worm belongs to

the genus Senga as new species i.e. Senga rupchandensis n.sp. The worm along

with intestine is utilized for the histopathological study.

On closer observation of the transverse section of healthy intestine of

Channa striatus (Bloch, 1793), the serosa, muscularis, sub-mucus and mucosa

layers of intestines were clearly observed which are intact and healthy (normal)

(Plate-20, Fig. 1) whereas in the infected intestine with the cestode Senga

rupchandensis n. sp. it is observed that the worm has high penetrative type of

scolex and they cause heavy mechanical tissue damage to their host. The colour

changes of the infected intestine from whitish to yellowish.

Histopathology caused by the parasite pierced to the intestinal tissue

makes the hole or vacuolization with the help of scolex.

In the transverse section of infected intestine with the Senga

rupchandensis n. sp. was clearly observed that, the worm penetrates through the

intestinal villi and ruptured the mucosa and sub-mucosal layers. The parasite

penetrates deeply through the intestinal tissue layer and causes heavy mechanical

injury by the penetrative type of scolex ((Fig. 2). The worm tried to obtain all the

nourishment from the host. The effect of parasites on the get serious of change

which ultimately reduces the absorption rate and interrupts the other metabolic

processes. The biochemical studies show the changes that occur in the infected

tissues including the change of carbohydrates, lipid, protein nucleic acid and

enzyme levels.

The present study showing that, the worm Senga rupchandensis n. sp.is

having penetrative type of scolex and is highly pathogenic parasite to the host,

Channa striatus (Bloch, 1793). This pathological result is resemblance and

discussed with Satpute and Agarwal, (1974a) also noticed inflammatory response

in submucosa and serosa of Clarias batrachus infected with Lytocestus indicus

and Diphyllobothrium penetra.

Karanis and Taraschewski, (1993), described the histopathological

changes in cyprinids infected by Caryophyllaeus laticeps, the scolex of

Caryphyllaeus laticeps cause only local compression of the hosts gut epithelium

and at the site of attachment of these cestodes vacuolation of epithelial cells and

ruptures the brush border.

Hiware, C.J. (2002) has studied the pathological changes of intestine of

Clarias batrachus infected by the Lytocestus maldurgensis, the scolex deep

penetration through the intestinal villi, totally breaks or damage the sub-mucosal,

mucosal, muscularis. The scolex parasite comes to lie near the serosa layer.

Molnar, K. (2003) has studied the histopathological changes in common

carp, Cyprinus carpio (L.) infected with Atractolytocestus huronensis and shown

that the cephalic part and neck of the Atractolytocestus huronensis is found in the

deeper layer of the intestinal mucosa, the scolex was surrounded only by the

basement membrane which separated it from the lamina propria of the intestinal

mucosa, worm penetrating into the deep layer of the mucosa containing intestinal

crypts and damage of the epithelium layer.

Molnar, K. (2005) shown that the cestode parasite, Neogryporhynchus

cheilancristrotus infected to Gibel carp fish. Cestode parasite is a truly

pathogenic species causing degeneration and inflammation in the intestinal wall,

their scolices of Neogryporhynchus cheilancristrotus intrudes into deeper layer

of the gut and breaking through the epithelium is located in the lamina propria of

the mucosa layer.


On closer observation of the transverse section of normal (healthy) liver of

the Mastacembelus armatus (Lecepede, 1800) was clearly observed in the

microscope (Plate-21, Fig.1) whereas in the infected liver with cestode

Circumoncobotrium jadhavae n.sp. cysts damage the liver tissue. In the infected

liver change the color from reddish to yellowish.

Histological examination of the infected liver with Circumoncobothrium

jadhavae n.sp. cysts is attached into the middle portion of the host liver. This

cysts is double layered with coiled the larvae in the cysts. The macrophages

gathered around the cyst wall, the necrosis of parenchyma cells immediately

around the cysts is evident and also large number of inflammatory cells around

the cyst, blockage of bile passages, enlargement of hepatocytes and liver

vacuolation from the cyst, the sinusoid were ruptured and filled with blood

(Fig.2).

The changes that are caused in the host livers are due to the mechanical

damage that may be due to the release of toxins by the parasites. The parasite not

only change the morphology of the host, but also interfere with the nutrition and

metabolism and disturb the movements and secretary functions of the alimentary

canal and its associated glands, the influence of the parasite on the host is

adverse.

The present study showing that, the histopathology of infected liver with

Circumoncobothrium jadhavae n.sp. cyst. This results are matching in

accordance with the study carried out by Raymond C. Bowen, (1969) have

studied the histopathology of plerocercoid of protocephalus sp. infected to the

liver of Lepomis macrochirus. Each larval tapeworm was surrounded by a

cellular capsule containing spindle shaped cells with elongated nuclei. Liver cells

adjacent to the capsule appeared more vacuolated than of the parenchyma. Paul

C. Stromberg and John L. Crites, (1974) has studied the histopathological

changes in liver of the fish Marone chrysops infected by the larval

Trianenophorus nodulosus. A distinct cellular response in the liver, destruction

of the proximal liver parenchyma, compression pancreatic tissue, destruction of

the squamous, metaplasia and fibrosis. S. Radhakrishnan et al., (1983) has

studied the histopathology of marine teleost fish, Saurida tumbil (Bloch) liver

infected with cestode Penetrocephalus ganapati cyst. Scolex was embedded and

encysted in the liver and neck connecting the scolex with strobila its major part

lies in the viscera, the ensheathed neck separate and take independent paths so

that the scolices are encysted seperatly inside the liver.

Ewa Dzika et al., (2005) have studied the histopathology of the liver of

Ratilus ratilus L. infected with the Paradilepis scolecina, resulted changes in the

liver, vacuolar degeneration of liver cells, dispersed foci in liver parenchyma,

large melano-macrophage centre and necrosis was accompanied by focal

infiltration of lymphoid cells.


On closer observation of the transverse section of healthy intestine of host,

Mastacembelus armatus (Lecepede, 1800) all layers are clearly observed (Plate-

22, Fig.1), whereas in the infected intestine with trematode parasite,

Genarchopsis paithanensis n. sp. causing damage to the epithelial layer.

Genarchopsis paithanensis n. sp. trematode is found in the anterior part of

the intestine. Longitudinal section of the trematode parasite infected with the

intestine of Mastacembelus armatus, the anterior end of the trematode parasite

Genarchopsis paithanensis n.sp. was approaching the intestinal villi (Fig.2) and

damage the epithelial layer, embedded in the fibroblast, lymphocytes, plasma

cells and attached to the intestinal villi, therefore, causing inflammation,

vacuolation and damage the intestinal villi (Fig.3).

The worm is not only successful to enter into the intestine forming the

ulceration in the intestinal wall causing damage to the host tissue but the parasite

may affect host physiology in many ways that induce stress in the host. The

parasitic infection to the disturb the circulation of sugar levels which in turns

effects other metabolic pathways.

The present study showing that, the Genarchopsis paithanensis n.sp.

damage the epithelial layer, this result are matching in accordance with the

studies carried out by Barbara, (1980) who has studied the pathological changes

brought by Bucephalus polymorphus and Rhipidocotyle illensis in cyprinid fry.

The damage caused to a white sucker fish by the trematode parasite

Triganodistomum attenuation was reported by Muzzal, (1980). In contrast,

Banergee G. et al., (2006), has studied the histopathology of the intestine of

freshwater fish, Clarias batrachus infected by the trematode parasite,

Genarchopsis goppo caused extensive damage to various layer of the intestine

right from mucous membrane to muscularis layer, extensive damaged caused to

the villi.


On closer observation of the transverse section of normal (healthy) liver of

Mastacembelus armatus (Lecepede, 1800), the liver cell, hepatocytes are clearly

observed (Plate-23, Fig.1), whereas in the infected liver with the trematode

Allocreadium aurangabadensis n.sp. cysts damage the liver tissue, vacuolization

and change the shape and size of liver.

Histopathological examination of infected liver with Allocreadium khami

n.sp. trematode shows that cysts were attached to the serosal coat of the liver and

damage the liver tissue. The cyst is large with parasite and filled with eggs. The

macrophages scattered around the cysts, distinct shape of hepatocytes,

vacuolization and damage the liver cell or necrosis is seen with (Fig.2), sinusoid

were ruptured. The infection in the liver by a parasite causes disturbances in the

vital functions of the glands. These disturbances may directly affect the chemical

nature of the infected tissue by lowering or increasing the important molecules

which plays important role in metabolism.

Liver is the one of the important digestive glands. It is the chief organ of

the process of detoxification. It plays a role in the metabolism of carbohydrate,

protein and lipid, storage of glycogen, desaturation of fatty acid, amino acid

carried out by the liver (B.Lakshman Reddy, 2006).

The result of the present study are similar in accordance with the studies

carried out A.J. Mitchell, (1982) who studied the histopathology of the liver of

Fathed minnow (Pinephales promeles) fish infected with Posthodiplostomum m.

minimum trematode encysted in liver tissue causing little damage to hepatocytes,

melanin-macrophage centres were diffusely scattered throughout the fibrinous,

fibroblast produce containing collagenous connective tissues. Reddy B. Lakshma

et al., (2006) have studied the histopathological in liver of freshwater fish,

Channa punctatus infected with the Euclinstomum heterostomum includes

enlargement of hepatocytes, vacuolation of cytoplasm, disarray of hepatochord,

hypertrophy of hepatocytes and liver vacuolation. The present study is in

contrast from result of J.E. Revenga et al., (2006) have studies and observation

of the histopathology of the liver of Galaxias maculates fish infected parasites

found in the hepatic parenchyma of “puyenes”, causing hepatic parenchyma was

unaltered a distance from the foci of the lesions and weak inflammatory

reactions.


On closer observation of the transverse section of healthy

buccopharyngeal tissue of the host, Channa striatus (Bloch, 1793) it located

posterior region of the head are clearly seen (Plate-24, Fig.1), whereas

buccopharyngeal area infected with trematode Orientocreadium striatusae n.sp.

cysts embedded between the pharynx and gills causing inflammation of

buccopharyngeal wall (Fig.2). Externally observed, changes the colour of

infected gills from reddish to whitish. Gills are the very important part for the

respiration in fishes.

In the transverse section of the infected buccopharyngeal area with

Orientocreadium striatusae n. sp. the trematode cysts is large and surrounded by

dense connective tissue, inflammation of the buccopharyngeal area. Parasite

causes breaks and destroys the fat tissue of the host. Blockage the gill capillaries,

lamellae and prevent the blood circulation as well as respiration of the host (Fig.

2, 3).

The present study showing that, the Orientocreadium striatusae n.sp.

damages the buccopharyngeal area of the host, Channa striatus (Bloch, 1793).

This result discussed with Wallace A. Evans, (1974) have seen the

histopathology of gill arches of Cutthrout trout (fingerlings) infected with blood

flukes, Sanguinicola klamathensis. These parasite eggs occurred more frequently

in the capillaries of gills, changes in gills, hyperplasia, and chronic inflammation

of the gill epithelium and fusion of lamellae. Baur O.N. et al., (1959) indicated

that the blocking of gill capillaries by Sanguinicola sp. eggs causes necrosis of

the gills. Schaperclaus, W. (1954) indicates that the eggs of Sanguinicola

intermis infected to the fish causing they clog of gill capillaries bring about

thrombosis and death the fish. Robert E. Olson et al., (1997) have studied the

histopathology of gills of Steelhead trout (Oncrynchus mykiss) infected with the

trematode metacercaria caused the hyperplasia of filament epithelium, in some

areas did lead to fusion of gills lamellae and loss of respiratory surface.

Jorge da Costa Eiras et al., (1999) have studied the histopathology of gill

arches infected with the Clinostomum marginatum cyst. In the gill arches the

dense connective tissue capsule surrounding the parasite causing the fat tissue

destroyed, its all completely broken, the typical architecture of the tissue was

changed into an irregular mass containing a great amount of cell remnant and

debris, tissue destruction from the cyst.

Jirawat Tudkaew et al., (2008) have studied the histopathology of gill of

the Orange spotted grouper (Epinephelus coioides Hamilton, 1822) it is marine

fish infected with Ganapodamium epinephili trematode cyst showed the parasites

were tightly packed underneath the mucosal layer of the primary gill lamellae,

secondary lamellae have disappeared, the cyst wall was composed of epithelial

cells of primary gill lamellae, parasitic cysts expanded and destroyed the

secondary lamellae and small suckers embedded underneath of the gill

epithelium.

CONCLUSION:

It is concluded that, the parasites are very dangerous for human being as

well as other animals. Fishes gives content of protein, provides vitamin A,

vitamin D and as a commercial economic point of view and useful for

preparation of soup, liver oil. The study of the helminth parasites are necessary

because the parasite affects on productivity of the fish population which may

cause deterioration in their food value, by decreasing growth rate, reducing the

quality of flesh, loss of protein, behavioral changes as well as reduces the

absorption and metabolic process result may in heavy mortality. Besides,

infected fishes act as a very potent source of helminth infections of man and they

transmitted (to man) only through eating of fish.

CHAPTER-IV DNA FINGERPRINTING

DNA FINGERPRINTING INTRODUCTION:

DNA is the molecule of hereditary; it is an integral component of all

living matter with the exception of RNA viruses. Any living or non living

organic matter containing relatively intact DNA fragment is used for DNA

typing analysis. A common source includes human or other animal blood, semen

and solid tissues and plant foliage and seeds.

DNA fingerprinting has become an indelible part of society helping to

prove innocence or guilt in criminal cases resolving immigration arguments and

clarifying paternity. “DNA fingerprinting started at the headquarters of the

British Antartic Survey in Cambridge,” says Professor Jeffrey, “ I collected a big

lump of seal meat from their lock up freezer and we got the seal myoglobin gene,

had a look at human myoglobin gene and there inside an intron in that gene was

tandem repeat DNA.

DNA fingerprinting has proved to be powerful in resolving genetic

identity or relationship and is applied in many diverse areas of biological

sciences including forensic science, paternity testing, animal breeding and

population genetics (Gill et al., 1985; Jeffrey et al., 1985). One attractive DNA

fingerprinting method is the detection of hyper variable simple repetitive DNA

by means of oligonucleotide probes, which make it possible to establish highly

informative DNA fingerprints for any eukaryotic organisms. It is useful for

identifying genetic relationship in domestic animals and wild birds (Buitkamp et

al., 1991). DNA fingerprinting use of genetic analyses, human percentage, rape

and homicide case in animal poaching and medical analysis.

The development of the molecular tools in the past decades has stimulated

systematic researches, nevertheless, work concerning the parasite is far from

being at the level of that of the hosts, especially as the use of molecular data for

the study of the phylogenetic relationship and the genetic characterization of the

population of parasites were largely limited to the species being of medical or

economic interest (Poulin and Morand, 2000). Although a pair number of genes

has been used to study parasitic phylogeny (Olsen and Tkach, 2005). The

parasite is more commonly encountered and sampled in the larval stage and

hence; morphological studies are not possible, as they are done on adult

reproductive complexes or fully developed plerocercoid moreover, few number

of morphological characteristics are available for species identification

(Dubinina, 1980). Many of the conventional biochemical identification

techniques such as, isoenzyme electrophoresis are being by DNA typing.

Okamoto et al., (1995) applied DNA fingerprinting with C (Ac)5 to

analysis of genetic variation within Taenia taeniformis and reported that C (Ac)5

was a highly resoluble and informative probe for cestodes.

Morphological study has shown that there can be significant variation in

parasite length of the adult of Oesophagostomum bifurcum (Nematode) among

species of primate host (Blotkamp et al., 1993). These observation have

suggested the existence of population variation within Oesophagostomum

bifurcum from human and non-primates, while previous investigations of

ribosomal and mitochondrial DNA did not reveal clear evidence of genetic sub

structuring within Oesophagostomum bifurcum from human and different species

of non-human primates (Gasser et al., 1999; De Gruijter et al., 2002), a recent

study using random amplification of polymorphic DNA (RADP) analysis

provided support for the existence of different genetic groups of

Oesophagostomum bifurcum according to host species (de Gruijter et al., 2004).

The fingerprinting of parasitic nematode Haemonchus contortus (Otsen et al.,

2001) and bovine lungworm, Dictyocautus viviparous (Huglund et al., 2004) and

no investigation of parasitic nematodes of human health importance.

The polymerase chain reaction (PCR) ISSR is a relatively novel technique

used to screen a large part of the genome without prior knowledge of sequences.

The method provides highly reproducible results and generates abundant

polymorphism in many systems. It is proven to be efficient in distinguish

between populations and closely related species (Zietkiewicz et al, 1994;

Robinson et al., 1997; Wolfe et al., 1998 a, b; Hundsdoerfer and Wink, 2006;

Maltgiati et al., 2006). The largest regions by ISSR yield higher polymorphism

and reducibility (Fang and Roose, 1997; Luque et al., 2002; Wu et al., 2005).

According to Behura, (2006) the ISSR technique represents one of the most

promising tools in population genetics and deserves increased attention

(Esselman et al., 1999). However the relevance of this approach for phylogenetic

studies and particularly when comparising genera, tribes or families (Simmons et

al., 2007).

The PCR method is commonly used to identify parasitic infection and

species diversity for helminthes. The application of PCR method were reported

to investigate the genetic variations of the global cestode, Taenia solium

(Okamoto et al., 2001), to detect the liver fluke, Ophisthorchis viverrini from

bithynid snail and cyprinid fishes (Maleewong et al., 2003) and to identify the

minute intestinal trematodes, Stellantchasmus falcatus, infecting half-beaked fish

(Dermogenus pusillus) (Sripalwit et al., 2003). Shiff et al., (2000) estimated the

quantity of genomic DNA of the cercarial-stage blood fluke, Schistosoma

haematobium infected snail found that 2.5 ng DNA was optimal for the RAPD

method.

DNA fingerprinting of Ophisthochis viverrini detected by specific primer

(OV-61 and OV-6R), the lowest DNA quantity was 2× 10-17 ng extracted from

only one eggs, which was taken directly from the fluke from patient faces

(Wongratanachewin et al., 2001). DNA fingerprinting of the lungs fluke,

Paragonimum heterotremus genomic DNA from 5 eggs taken from the faces of

an infected cat was used pPH-13 specific DNA probe and produced 100%

sensitivity (Intapal et al., 2005). The differences in species size and stages of

parasite may effect the quantities of genomic DNA used in PCR method, the

HAT-RAPD method (Anuatalabhochai et al., 2000) was used effectively to

identify Stellantchasmus falcatus in half-beaked fish by (Sripalwit et al., 2003).


n. sp., Circumoncobothrium jadhavae n.sp., Genarchopsis paithanensis n.sp.,


freshwater fishes namely Channa striatus (Bloch, 1793) and Mastacembelus



Buffer and chemicals:

Extraction buffer: 50 mM Tris-HCl, pH 8.0, containing 1% CTAB (hexadecyl trimethyl ammonium

bromide), 0.75 M NaCl, 10 mM EDTA and 100 mg/ml proteinase K.

TAE (Tris-Acetate-EDTA)

• Tris base 48.4 gm

• Acetic Acid11.42 ml

• 0.5 M EDTA 20 ml

Adjust pH 8.5using KOH and make final volume to 1 liter.

TBE (Tris-Borate-EDTA) (10X)

• Tris base 108 gm

• Boric Acid 55 gm

• Na4EDTA 9.3 gm

Adjust pH 8.3 using KOH and make final volume to 1 Liter.

DNA extraction method:

DNA was extracted from 05 different parasites according to standard

protocol Waldschmidt et al., (1997).

Five parasite such as Circumoncobothrium jadhavae n.sp., Genarchopsis

paithanensis n. sp., Allocreadium khami n.sp. from Mastacembelus armatus

(Lecepede, 1800) and Senga rupchandensis n. sp., Orientocreadium striatusae

n.sp. collected from Channa striatus (Bloch, 1793). Each parasite was ground

individually in a mortar and pestle containing liquid N2 and homogenized with

extraction buffer. Each parasite was ground in a micro centrifuge tube with a

plastic pestle in the presence of extraction buffer. The samples were then

incubated at 65oC for 30 min. Samples containing adult individuals were

deproteinized twice with one volume each of chloroform. Parasite homogenates

were then deproteinized once with one volume Phenol: Chloroform (1:1) and

once with one volume chloroform. After deproteinization the samples were

centrifuged at 11,750 g for 5 min. Nucleic acid was precipitated by addition of

one volume of cold isopropanol and incubation at -20oC for 2 to 24 h. After

centrifugation at 16,000 g for 30 min the pellets were washed twice with 70%

(v/v) ethanol and vacuum dried for approximately 15 min or dried in the air for

30 min. The pellet was resuspended in 200 ml TE (10 mM Tris-HCl, pH 8.0, 1

mM EDTA) and incubated with RNAse (100 mg/ml) for 30 min at 37oC. Dry the

DNA pellet and dissolve in TE buffer and dissolve the pellet. Store at 4oC.

A small aliquot of isolated DNA was run on a 1% (w/v) TAE gel to check

the quality of DNA Sample.

PCR amplification of DNA with ISSR primers:

ISSR primer sets were ordered from University of British Columbia

(UBC). 10 ISSR primers were used for initial screening. Out of 10 primers, 4

primers gave the amplification (in terms of repeatability, scorability and ability)

were selected for identification. The selected 4 primers were 14- 23 mers based

with various di-, tri-, nucleotide ISSR repeats. Following primers gave the

amplification. The primer sequences such as, Primer 811 – GAG AGA GAG

AGA GAG AC; Primer 812 –GAG AGA GAG AGA GAG AA; Primer 814 -

CTC TCT CTC TCT CTC TA and Primer 816 – CAC ACA CAC ACA CAC

AT.

PCR technique has promoted the development of a range of molecular

assay systems which detect polymorphism at molecular level. In this study we

used the most widely adopted PCR based ISSR marker technology for

characterizing the natural variation amongst the parasite isolates. PCR reactions

were carried out in a thermal cycler.

Preparation of master mix for PCR:

Taq buffer (10X) - 2.0µl

DNTP’s (10Mm) - 2.0µl

Taq polymerase (3U/ul) - 0.4µl

Tween 20 - 0.2µl

MQ water -3.4µl

Primer (3pmol/ ul) - 2.0µl

Template DNA -3.0µl

Setting up a PCR thermal cycler:

• 94 degree for 3 min. - hot start Denaturation

• 94 degree for 15 sec. – Denaturation (38 cycles)

• Annealing for 30 sec. – temp varies according to primer

• Set the reaction for 30 to 38 cycles.

• 72 degree for 1 minute – Extension (38 cycles)

• Last 72 degree for 5 minutes – final extension.

• Proceed 38 reactions

• After completing 38 cycles, load the sample on 1X TBE gel.

Post PCR processes:

PCR products were separated by electrophoresis using 1% Agarose gel in 1X

TBE buffer.

ELECTROPHORESIS:

Principle:

Electrophoresis is the movement of charged molecules under the electric

field. Electrophoresis is carried out by using agarose gels which are more porous.

It is used to separate large sized macromolecules like DNA or RNA. DNA is

negatively charged and move towards anode when an electric field is applied.

Requirement:

Agarose powder, boric acid, tris, EDTA, Distilled water, gel loading stock(

bromophenol blue), Running buffer commonly used are Tris –borate EDTA(

TBE) and Tris- acetate- EDTA (TAE)

Equipments:

Electrophoresis apparatus with power supply, magnetic stirrer, micro-pipettes,

eppendorf tube etc.

Procedure:

1. Dissolve 0.8 agarose in 100ml of 1X TBE by gentle heating on

magnetic stirrer with hot plate. This will result in 0.8% agarose gel.

Cool the contents.

2. Using a tape seal both the open sides of gel tray.

3. Insert the comb in such a way that 1mm gap between the teeth and

surface of tray could be made.

4. To get the thickness of gel to about 4-5 mm, pour agarose solution into

the tray. Now hold this for about 30-40 minutes.

5. Gently remove the comb when gel has been solidified. Remove the

tapes and keep gel tray into electrophoresis tank.

6. Pour TBE into the tank so that gel can be immersed by about 5mm.

7. Carefully load DNA samples into slots of submerged gel. The samples

must be settled at the bottom of the slot.

8. Connect the electric lead in such a way that a negative terminal should

be at the end where sample has been loaded.

9. Run electrophoresis at 60 V until loading dye bromophenol blue

migrate to the other end of the gel.

10. Turn the button off and disconnect the electric leads. Take out agarose

gel from the electrophoresis tank.

Gel staining and visualization:

A dye called Ethidium bromide is used to detect the DNA. It intercalates

with the DNA. Therefore location of DNA can be detected and visualized when

fluoresces under U.V light. From Ethidium bromide stock solution, 1mg/ ml is

dissolved in water the agarose gel taken out from electrophoresis tank and dipped

in Etbr solution for some time. PCR products on separation by 1% agarose gel

using TBE gel buffer and staining are visualized by ultraviolet illumination. The

separated genomic DNA as bands fluoresces under U.V light which makes its

presence positive.

Band scoring and data analysis:

For each sample, each fragment / band that was amplified using ISSR

primers was treated as a unit rearrangement in genome. The primers which were

given scorable and consistently reproducible amplicons were considered. The gel

pictures were taken and documented to computer by using Alpha Imager gel

documentation system and size of each amplicon was measured by using Alpha

Imager Software with respect to standard molecular weight DNA ladder and

molecular weight of each of the potential specific bands was calculated using the

software program Alpha Imager.

Phylogeny analysis:

The Dendrogram was plotted by using bioinformatics phylogeny Free

Tree and Tree View of DNA fingerprint analysis tool.

The phylogenetic analysis carried out by using Free Tree bioinformatics

phylogeny tool of DNA fingerprint analysis based on amplicons sizes given by

primers showed that isolate number 5 is the basic original sample. Bootstrapping

analysis was carried out to find the original an-sister progeny analysis amongst

these samples by using Free Tree bioinformatics tool of phylogeny analysis.

Dendogram analysis

Dendrogram was plotted y using the Free Tree bioinformatics software to

find out the evolution pattern.


The result from PCR based ISSR-RAPD method revealed that of the four

primers tested such as, Primer 811 – GAG AGA GAG AGA GAG AC; Primer

812 –GAG AGA GAG AGA GAG AA; Primer 814 - CTC TCT CTC TCT CTC

TA and Primer 816 – CAC ACA CAC ACA CAC AT produced DNA banding in

parasite, total genomic DNA extraction from cestode and trematode parasite such

as, Circumoncobothrium jadhavae n.sp., Genarchopsis paithanensis n.sp.,

Allocreadium khami n.sp. from Mastacembelus armatus (Lecepede, 1800) and

Senga rupchandensis n.sp., Orientocreadium striatusae n.sp. from the host,

channa striatus (Bloch, 1793).

Amplification of genomic DNA of five parasite sample using ISSR

marker analysis, yielded total amplified 50 fragments. ISSR analysis yielded

fragments that could be scored, out of which 26 were polymorphic while the

remaining were 24 monomorphic in nature.

In ISSR analysis number of amplified fragments ranged from 05 to 15 and

which varied in size from 170 bp. to 2230 bp. UBC (University of British

Columbia) Primer No. 811, 812, and 816 showed the lowest number of bands

while the Primer No. 814 produced the highest number of bands. Out of the 50

amplified 26 bands produced 52% were polymorphic in nature with an average

of 10.04 % polymorphic fragments per primer.

The primer based poly (AC at 3’) produced maximum number of bands

(15 bands) while the primers having poly (AA at 3’) produced minimum number

of bands (5 bands). The PCR amplification products. The complete data was

based on a total of 50 bands.

A dendogram analysis was carried out by analysis bioinformatics

phylogeny tool Free Tree and Free view of DNA fingerprints anlysis. Distance

matrix was calculated by using Neighbour-joining tree construction method of

Nei and Li, 1979 which ranged from 0.03797 to 0.30556. five parasites were

clustered into four major clusters. Cluster-I comprises of sample Senga

rupchandensis n.sp., Genarchopsis paithanensis n.sp.; Cluster-II of single

parasite, Circumoncobothrium jadhavae n.sp.; Cluster-III of single parasite

namely Orientocreadium striatusae n.sp., Cluster-IV of single parasite namely

Allocreadium khami n.sp. thus, all five parasites are grouped together in Cluster

I, II, III, and IV. In present molecular analysis, all samples which are

significantly different from each other showed genetic variability.

The result of the present study are matching in accordance with the

studied carried out by the previous report also used Primer OPA-O9 out of the

four tested Primer to identify the hetrophyid trematodes and Ophisthorchis

viverrini (Sripalwit et al., 2003) and Metraginimus spp. (Yu et al., 1997 a, b).

The lowest DNA concentration and produced distinct bands for DNA analysis

between -1×10 -11 ng.

Li and Liao, (2003 ) after suggested for Digramma (a sister genus of

Ligula), for this low genetic variability is the migration of the definitive hosts of

Ligula (e.g. Larus risidibundus, Ardes cinnerca and Phalacrocorax carbo).

Indeed, it is was recently accepted that migratory birds transport organisms

across vast distances (McCoy et al., 2003; Gittenberger et al., 2006).

Chalbol Wongswad et al., (2006) observed the banding pattern of some

trematode used PCR based HAT- RAPD method revealed that, four primer

tested, only used OPA-04, OPA-08, OPA-09 Primer, produced DNA banding in

every trematode. OPA-09 was selected for further investigation, since its banding

pattern were most distinct.

Pheravut Wongsawad and Chalobol Wongsawad (2007) studied the DNA

fingerprinting of some trematodes ie., Stellantchasmus falcatus and Haplorchis

taichui and metacercarial stages used HAT-RAPD method with eight artibitory

primers OPA-02, OPA-08, OPA-9, OPN-02, OPN-03, OPN-09, OPX-13, OPH-

19 and observed the DNA banding pattern of both adult and metacercarial stage

of the trematodes. A total of 180 bands were examined as molecular weight by

Kodak ID Image between 160-2,865 bps and three bands were observed in all

trematodes of the both stages by OPA-02, OPA-08, and OPX-13 with molecular

weights of 580, 560 and 650 bps. The OPN-02 primer had the highest band

number (36) and produced the high specific band number (32) in all trematodes.

McGarry et al., (2007) used PCR based two set such as (set-I, set-II). In

primer-I, a product of 391 bp. was generated from the genomic DNA of Fasciola

hepatica whereas no product generated from DNA of Fasciola gigantica. PCR

based two primer set consistently amplified a 235 bp. product from the DNA of

both trematode parasite. Fasciola hepatica can be identified by the generation of

a 391 bp. by using primer set I and Fasciola gigantica can be identified by the

lack of a 391- bp product with Primer set-I and the presence of a 235 bp product

with primer set-II.

Bouzid W. et al., (2008) are observed the DNA banding pattern of 159

Ligula intestinalis sample from the different fishes , PCR based ISSR analyses

was performed using nine Primers, use only four Primers such as AUS1-AUS9,

TU1, CN1-CN3, FR1- FR3. the size of the bands displayed ranged from 250 to

1500 bp. This four Primers provided different patterns and number of bands but

gave almost the same percentage of polymorphism. The total polymorphism (P)

scored between population from different geographical regions was 100%

whereas less polymorphism was detected among inmdiviuals within each groups.

Indiviuals from China showed the highest polymorphism (55.45%), Germony

showed the lowest rate (4055%) the total gene diversity (Ht) was 0.293±0.019,

gene diversity within populations (Hs) was 0.065±0.003 and the global

coefficient of gene differenciation (Gst) was 0.776. European group gene

diversity nwas 0.230. Nei’s genetic indentity (I) ranged from 0.525 to 0.999. the

distances in the European-Tunisian group varied from 0,0002 between French

and Czech population to 0.0294 between German and Russian ones. Genetic

distances between European, Chinese, Algerian, Australian and Canadian. Five

main Cluster were defined fro 10 population. The MP tree showed a divergence

of groups, with a clear differenciation between analysed population from China,

Australia, Canada and Algeria.

CONCLUSION:

It is concluded that, today the study of DNA fingerprinting method is a

very important, this technique applicable for identification of individual, genetic

variation, identification of rape suspects, unknown murder, identify of criminals,

identify of burnt or unidentified dead body, it also helps in reuniting the lost

children with their respective parent or vise versa. DNA fingerprinting method

has been patent and being used in Europe and America and accepted in most

courts in the United States.

SUMMARY

CHAPTER-I:

The taxonomical study of the cestode and trematode parasites collected

from the freshwater fishes, namely Mastacembelus armatus (Lecepede, 1800)

and Channa striatus (Bloch, 1793). The cestode parasite belongs to the

Eucestoda, Order- Pseudophyllidea, Family- Ptychobothridae. From Order-

Pseudophyllidae one genera Circumoncobothrium as one new species described

Circumoncobothrium jadhavae n.sp. from Mastacembelus armatus and one

genera Senga as two new species are described Senga rambaei n.sp. from

Mastacembelus armatus and Senga rupchandensis n.sp. from Channa striatus

(Bloch, 1793).

The trematode belongs to the family-Azygiidae belongs one genera Azygia

described one redescribed species Azygia stunkardi Rai, 1964 from Channa

striatus; family Hemiuridae belongs one genera Genarchopsis described one new

species Genarchopsis paithanensis n.sp. from Mastacembelus armatus; family

Gorgoderidae, one genera Phyllodistomum described one species

Phyllodistomum aurangabadensis n.sp. from Channa striatus; family

Allocreadiidae, one genera Allocreadium described two new species , one species

Allocreadium khami n.sp. from Mastacembelus armatus and one species

Orientocreadium striatusae n.sp. from Channa striatus.

CHAPTER-II:

In this chapter deals with the study of histochemical analysis of acid and

alkaline phosphatase of cestode and trematode parasite from freshwater fishes

namely Mastacembelus armatus (Lecepede, 1800) and Channa striatus (Bloch,

1793).

Senga rupchandensis n.sp., Orientocreadium striatusae n.sp. collected

from Channa striatus and Circumoncobothrium jadhavae n.sp.; Genarchopsis

paithanensis n.sp.; Allocreadium khami n.sp. collected from Mastacembelus

armatus.

From histochemically observations of both acid and alkaline phosphatase

enzyme activity in Senga rupchandensis n.sp. and Circumoncobothrium

jadhavae n.sp. The enzyme activity are high in reproductive organ such as,

testes, cirrus pouch, ovary, vitellaria and egg shell and less activity in

musculature. In trematode, Genarchopsis paithanensis n.sp. high alkaline

phosphatase enzyme activity in sucker, reproductive organ and less in cirrus

pouch, musculature and acid phosphatase enzyme activity was observe in

reproductive organ except cirrus pouch; Allocreadium khami n.sp. high alkaline

phosphatase enzyme activity in intestinal caeca, uterus, vitellaria, testes;

moderate in ovary and acid phosphatase enzyme activity in reproductive organ;

less in cirrus pouch, sucker and musculature ; Orientocreadium striatusae n.sp.,

the high alkaline phosphatase activity in sucker, reproductive organ and acid

phosphatase enzyme activity high in reproductive organ, moderate in intestinal

caeca.

CHAPTER-III:

The histopathological study of the cestode and trematode parasites

collected from the freshwater fishes, namely Channa striatus (Bloch, 1793) and

Mastacembelus armatus (Lecepede, 1800)

Transverse section of the healthy intestine of host, Channa striatus

showed the healthy structure whereas in infected intestine with Senga

rupchandensis n.sp. is having penetrative type of scolex and they cause heavy

mechanical damage to the mucosa and sub mucosa layer.

Transverse section of the healthy liver of Mastacembelus armatus was

clearly observed whereas histopathological examination of the infected liver with

Circumoncobothrium jadhavae n.sp. cysts is attached into the middle portion of

the host liver, the necrosis of parenchyma cells immediately around the cysts is

evident and also large number of inflammatory cells around the cyst, blockage of

bile passages, enlargement of hepatocytes and liver vacuolation from the cyst,

the sinusoid were ruptured and filled with blood.

Transverse section of the healthy intestine of Mastacembelus armatus was

clearly observed whereas in the infected intestine with trematode parasite,

Genarchopsis paithanensis n. sp. causing damaged the epithelial layer and

approaching the intestinal villi, embedded in the fibroblast, lymphocytes, plasma

cells and attached to the intestinal villi, therefore, causing inflammation,

vacuolation and damage the intestinal villi.

Transverse section of the healthy liver of Mastacembelus armatus was

clearly observed whereas histopathological examination of the infected liver with

Allocreadium khami n.sp. cysts attached to the serosal coat of liver therefore,

vacuolization, change the shape and size of liver and damage the liver tissue.

Histological structure of healthy buccopharyngeal tissue of the host,

Channa striatus (Bloch, 1793) was clearly observed whereas histopathological

observation of buccopharyngeal tissue infected with the Orientocreadium

striatusae n.sp. attached between pharynx and gills causing inflammation, blocks

the gill lamellae and blood capillaries.

The worm is not only successful to enter into the intestine forming the

ulceration in the intestinal wall causing damage to the host tissue but the parasite

may affect host physiology in many ways that induce stress in the host. The

parasitic infection to the disturb the circulation of sugar levels which in turns

effects other metabolic pathways.

CHAPTER-IV:

In this chapter deals with the study of DNA fingerprinting of five parasite

sample, Circumoncobothrium jadhavae n.sp., Genarchopsis paithanensis n.sp.,

Allocreadium khami n.sp. collected from the freshwater fish Mastacembelus

armatus (Lecepede, 1800) and Senga rupchandensis n.sp., Orientocreadium

striatusae n.sp. collected from Channa striatus (Bloch, 1793). For DNA

fingerprinting using the four ISSR primers like, Primer 811 – GAG AGA GAG

AGA GAG AC; Primer 812 –GAG AGA GAG AGA GAG AA; Primer 814 -

CTC TCT CTC TCT CTC TA and Primer 816 – CAC ACA CAC ACA CAC

AT.

In ISSR analysis number of amplified fragments ranged from 05 to 15 and

which varied in size from 170 bp. to 2230 bp. UBC (University of British

Columbia) Primer No. 811, 812, and 816 showed the lowest number of bands

while the Primer No. 814 produced the highest number of bands.

Today, this technique is advanced and very important for identification of

individuals, genetic analyses, human percentage, rape and homicide case in

animal poaching and medical analysis.

SYSTEMATIC POSITION OF CESTODE PARASITES WITH THEIR HOSTS

PARASITES: HOSTS:

Class Eucestoda

Wardle, McLeod &

Radinoky, 1974

Order Pseudophyllidea

Carus, 1863

Family Ptychobethriidae

Luhe, 1902

Genus Senga

Dollfus,1934 Channa striatus

Species Senga rupchandensis n.sp. (Bloch, 1793)

Class Eucestoda

Wardle, McLeod &

Radinoky, 1974


Carus, 1863


Luhe, 1902

Genus Senga Mastacembelus

Dollfus,1934 armatus

Species Senga rambaei n.sp. (Lecepede,1800)

Class Eucestoda

Wardle, McLeod &

Radinoky, 1974


Carus, 1863


Luhe, 1902

Genus Circumoncobothrium Mastacembelus

Shinde, 1968 armatus

Species C. jadhavae n.sp. (Lecepede, 1800)

Class Trematoda

Rudolphi, 1808

Order Digenea

van Beneden, 1858

Family Azygiidae

Odhner, 1911

Sub family Azygiinae

Luhe, 1909

Genus Azygia

Looss, 1899

Species Azygia stunkardi Rai, Channa striatus

1964 (Redescribed) (Bloch, 1793)

Class Trematoda

Rudolphi, 1808

Order Digenea

van Beneden, 1858

Family Hemiuridae

Luhe, 1901

Sub family Halipeginae

Ejsmont, 1931

Genus Genarchopsis

Ozaki, 1925 syn. Progonus

Looss, 1899; Preoccupied

Genarches Looss, 1902;

Preoccupied Ophiocorchis Mastacembelus

Srivastava, 1933. armatus

Species G. paithanensis n.sp. (Lecepede, 1800)

Class Trematoda

Rudolphi, 1808

Order Digenea

van Beneden, 1858

Family Gorgoderidae

Looss, 1901

Sub family Phyllodistominae

(Nybelin, 1926)Yamaguti,

1958

Genus Phyllodistomum Braun, 1899 Channa Striatus

Species P. aurangabadensis n.sp. (Bloch, 1793)

Class Trematoda

Rudolphi, 1808

Order Digenea

van Beneden, 1858

Family Allocreadiidae

(Looss, 1902) Stossich, 1903

Sub family Allocreadiinae

Looss, 1902 Mastacembelus

Genus Allocreadium Looss, 1900 armatus

Species A. Khami n.sp. (Lecepede, 1800)

Class Trematoda

Rudolphi, 1808

Order Digenea

van Beneden, 1858

Family Allocreadiidae

(Looss, 1902) Stossich, 1903

Sub family Orientocreadiinae

Yamaguti, 1958

Genus Orientocreadium

Tubangui, 1931 Syn. Ganada

Chatterji, 1933; Neoganada Dayal,

1938;Nizamia Dayal, 1938;

Ganadotrema Dayal, 1949;

Paratormopsolus Dubinina et

Bychowsky in Skrjabin, 1954;

Macrotrema Gupta, 1951. Channa striatus

Species O. striatusae n.sp. (Bloch, 1793)

SYSTEMATIC POSITION OF THE HOST

HOSTS Series Pisces

Class Actinopterygii

Order Periformes

Family Channidae

Genus Channa

Species C. striatus

(Bloch, 1793)

Series Pisces

Class Teleistomi

Order Mastacembeleformes

Family Mastacembellidae

Genus Mastacembelus

Species M. armatus

(Lecepede, 1800)

REFERENCES

Agarwal, V. (1964): Studies on some new trematodes from freshwater fishes of

Lucknow. Ind. J. Helm. 16 (2): 82-89.

Akhtar, N. (1986): Potential fishes in Pakistan, P.A.R.C. News. Monthly

Newsletter of Pakistan. Agri. Res. Counsil, 1-2 & 8.

Amlacher, E. (1961): Taschen buch der fish Krankheiten: Jena: Gustav Fischer

Anuntalabhochai, S., Chiangda, J., Chundet, R., Apavat, P. (2000): Genetic

diversity within lychee (Litchi Chinesis Soonn.) based on RAPD

analysis. [Abstract] Cairns, Australia: International symposium on

Tropical and subtropical Fruits. 26th November-1st December; 45.

Araxie Kilejian, Lewis A. Schinazi, Calvin W. Schwabe (1961): Host-parasite

relationships in Echinococcosis. V. Histochemical observations in

Echinococcus granulossus. The Journal of Parasitology, Vol. 47,

No. 2: 181-188.

Arme, C. (1966): Histochemical and Biochemical studies on some enzymes of

Ligula intestinalis (Cestoda: pseudophyllidea). The Journals of

Parasitology, Vol. 52, No. 1, 63-68.

Arme, C. and Read, C.P. (1970): A surface enzyme in Hymenolepis diminuta

(Cestoda). J. Parasit. 56, 514-516.

Bailey, W.S. (1951): Host tissue reactions to initial superimposed infection with

Hymenolepis nana. Var Fraterna. J. Parasit, 37: 440-444.

Banerjee, G., Lakshma Reddy B., and Bikshapthi, V. (2006): Histopathology

and histochemistry of the intestine of Clarias batrachus due to

trematode, Genarchopsis goppo (Ozaki, 1925). J. Aqua. Biol., Vol.

21 (2): 257-262.

Barbara Baturo (1980): Pathological changes in Cyprinid fry infected by

Bucephalus polymorphus and Rhipidocotyle illensis metacercariae

(Trematoda, Bucephalidae). Act. Parasitol. Pol, 27 (15-18): 241-

246.

Barry, D.H. and Mawdesley-Thomas, L.E. (1968): Enzyme histochemistry of

the adult liver fluke, Fasciola hepatica. Experimental

Parasitology, 23: 355-360.

Bashirullah A.K.M. and Mustak Elahi (1972): On two new species of

Genarchopsis Ozaki, 1925 from freshwater fishes of Dacca,

Bangladesh. Riv. Di. Parasit. 33 (4): 277-280.

Baur, O.N. (1959): The ecology of parasites of fresh water Fish: In: Parasites of

fresh water fish and biological basis of their control and river

fisheries, 45: 3-215.

Baur, O.N., Raikovo, K.V., Abrosov, V.N., Zaika, V.E. and Kheisin, E.M.

(1959): Parasite of freshwater fish and biological basis for their

control. Bull. State Sci. Res. Tnst. of Lake and River Fisheries,

Vol. 49 (English transl. Office Tech. Services, V.S. Dep.

Commerce, Washington, D.C., 1962. TT61-31056, 236 p) 243-

248.

Becejac, S. and Krvavica, S. (1964): On the presence and localization of

alkaline phosphatase activity in liver fluke (Fasciola hepatica L.).

Veternarski Arhiv Tugoslavia, 34: 56-59.

Behura, S.K. (2006): Molecular marker systems in insects: Current Trends and

future avenues. Mol Ecol. 15: 3087-3113.

Bhadauria, S., Dandotia, M.R. (1984): Studies on the trematode parasite of

freshwater fishes with special reference to Gwalior region. Part II.

On new genus and some unknown and known species. Riv. Di

Parasitologia, Vol. 1 (XLV): 349-383.

Bhalerao, G.D. (1937): Studies on the Helminths of India, Trematoda IV.’ Jour.

Helminth., 15.

Bhargavi, G.Y. and Krishna, G.V.R. (1993): Haematological changes in fresh

water fish, Channa punctatus (Bloch) infected with metacercariae

of progenetic worm Euclinostomum heterostomom (Trematoda).

Proc. Indian. Sci. Cong. Ass. Goa. (abs), 80 pt IV, sect-VIII, 174-

175.

Bhure, D.B., Padwal, N.D. and Jadhav, B.V. (2007): A new tapewarm Senga

jadhavae n.sp. (Cestoda: Pseudophyllidae) from Mastacembelus

armatus, Aurangabad (M.S.) India. Proc. Zool. Soc. of India. Vol.

6, No.2: 45-52.

Bijukumar, A. (1996): Nematode parasites associated with flatfishes (Order:

Pleronectiformes) off the Kerala Coast. J. Mar. Biol. Assoc. India,

38: 34-39.

Blotkamp, J., Krepel, B.P., Baeta, S., van’T Noordende, J.M. and

Polderman, A.M. (1993): Observations on the morphology of

adult and larval stages of Oesophagostomum sp. isolated from man

in northern Togo and Ghana. Jounal of Helminthology 67: 49-61.

Bogitsh, B.J. (1963): Histochemical studies on Hymenolepis microstoma

(Cestoda: Hymenolepididae). J. Parasit. 49: 989-997.

Borde, S.N.and Jawale, S. (2008): A new species of Ptychobothridae from a

fresh water fish in Marathwada region (M.S.). National Journal of

Life Sciences. 5 (3): 121-124.

Bose and Sinha (1979): The histopathology of the stomach wall of the fish

Channa gachua (Ham) (Channidae) attributable to the digenetic

trematode, Genarchopsis goppo (Ozaki) Hemuridae, current

science 48 (16): 747-748.

Bose, K. C. and Sinha, A. K. (1981): Histopathology of Clarias batrachus

(Linn.) infected by Lytocestus indicus Moghe, 1925. Science

culture 47: 186-187.

Bouzid, W., Lek, S., Mace, M., Hassine Ben O., Etienne, Legal, L and Loot,

G. (2008): Genetic diversity of Ligula intestinalis (Cestoda:

Diphyllibothriidea) based on analysis of inter-simple sequence

repeat markers. J. Zool. Syst. Res. 46 (40): 289-296.

Braun, M. (1899): Trematoden der Dhal’schen Sammlung aus Neu Guinea.

Centr. Bakt. Parasit. Abt., 25: 729.

Buitkamp, J., Ammer, H. and Geldermann, H. (1991): DNA fingerprinting in

domestic animals. Electrophoresis 12: 169-174.

Byochowsky, B.E., and Dubinina, M.N. (1954): Materials on the systematics of

digenetic trematodes of the family Acanthoppidae Luhe, 1909.

Zool. Zhur. 33: 188-193.

Chalbol Wongsawad, Pheravut Wongsawad, Jong-Yil Chai, Thipmani

Paratasilpin and Somboon Anuntalabhochai (2006): DNA

quantities and qualities from various stages of some trematodes

using optical and HAT-RAPD methods. Department of biology,

Faculty of Science, Chiang Mai University, Chiang Mai 50202,

Thailand. Vol. 37 (suppl 3): 62-68.

Chandra and Banerjee (1993): Digenetic trematodes from freshwater fishes of

Mymensingh families Clinostidae and Hemiuridae. Rivista Di

Parasitologia Vol-X (LIV)-N. 1: 81-91.

Chandra, K.J. (1994): Infection, concurrent infections and fecundity of

Procamallanus heteropneustes Ali parasite to the fish,

Heteropneustes fossilis. Enviro. and Ecol., 12: 679-684.

Chandra, K.J. and Modak, P.C. (1995): Activity, agining and penetration of

first larvae of Procamallanus heteropneustes. Asian Fish, Sci., 8:

95-101.

Chatterji, R.C. (1933): On the trematode parasites of a Rangan siluroid fish,

Clarias batrachus (Linnaeus, 1785). Bull. Acad. Sci. U.P. Agra &

Oudh, India 3: 33-40.

Chincholkar, L.N. and Shinde, G.B. (1976): Two new species of

Circumoncobothrium Shinde, 1968 (Cestoda: Pseudophyllidea)

from a freshwater fish in India. Marath. Univ. J. Sci., XVI (Sci.

No. 9): 183-185.

Chishti M. Z. and Bakshi S. (1992): New species of the genus

Pomphorhynchus (Monticeli 1905) from Cyprinus carpio

communis. Current trends in Fish and fishery biology and Aquatic

Ecology. [Ed. Yousuf, Raina and Qadri] XV +365.

Chishti, M.Z., Feroz, A.S., Fayaz, A. (2002): Histopathological study of

Pomphorhynchus in fishes of Kashmir. 16th National Congress of

Parasitology. Barelli, India. 176.

Chishti, M. Z., Feroz, A. S., Mahboob, H. (2003): Study of humoral immune

response of fishes to helminth infection. Journal of Parasitic

Diseases. 3 (6): 54-59.

Christina Ohman-James (1968): Histochemical studies of the cestode

Diphyllobothrium dentriticum Nitzsch, 1824. Z.F. Parasitenkunde,

30: 40-56.

Christina Sommerville (1982): The pathology of Haplorchis punilio infections

in cultured tilapias. J. Fish Dis., J (3): 243-250.

Chung, Yui-tan (1981): A study on the histopathology in the walfian ducts of

Hypentelium nigricans (Osteichthyes: catostomidae) caused by

Phyllodistomum superbum Q.J.Taiwanmus (Taipeli), 34 (3/4):

237-240.

Coleman, R.M. and DE. Sa, L.M. (1964): Host response to implanted adult

Hymenolepis nana. J. Parasit., 50 (suppl.) 17.

Cooper, A.R. (1915): Trematodes from marine and freshwater fishes, including

one species of ectoparasite tubellarian. Trans. Roy. Soc. Can. Sect.

IV , 9: 181-205.

Cross, S.X. (1933): Some host parasite relationship of the Trout Lake region of

Northen Wisconsin. J. Parasitol., 20: 132-133.

Dayal, J. (1938): “ A new trematode, Phyllochorus macronium N.G.N. sp.

belonging to the family Gorgoderidae Looss (1901) from the body

cavity of freshwater fish, Macrones tengara.” Proc. Ind. Acad.

Sci., 13, 7: 138-142.

Dayal, J. (1938): Studies on the trematode parasites of fishes. A new trematode

Neoganada barakankiae (nov. gen., nov. spec.) from Clarias

batrachus. Proc. Indian Acad. Sci., Sec. B7: 132-137.

Dayal, J. (1938b): Studies on the trematode parasites of fishes. A new trematode

Nizamia hyderabadi n. gen., n. sp. from intestine of freshwater

fish, Ophiocephalus punctatus. Proc. Nat. Acad. Sci. India, Sec .

8: 53-58.

Dayal, J. (1949): Trematode parasites of Indian fishes, Part II. Indian J. Helm. 1:

93-120.

De Gruijter, J.M., Polderman, A.M., Zhu, X.Q. and Gasser, R.B. (2002):

Screening for haplotypic variability within Oesophagustomum

bifurcum (Nematoda) employed a single-strand conformation

polymorphism approach. Molecular and cellular probes 16: 185-

190.

De Gruijter, J.M., Ziem, J., Verweji, J.J., Polderman, A.M., and Gasser,

R.B. (2004): Genetic substructing within Oesophagustomum

bifurcum (Nematoda) from human and non-human primates from

Ghana based on random amplication of polymorphic DNA

analysis. American Journal of Tropical Medicine and Hygiene 71:

227-233.

De, N.C. and Maity, R.N. (2000): Development of Procamallanus

saccobranchi (Nematoda: Camallanidae), a parasite of a

freshwater fish in India. Folia. Parasitol., 47: 216-226.

Deshmukh, R.A. and Shinde, G.B. (1980): On Senga khami (Cestoda:

Ptychobothridae) from the freshwater fish. Indian Jour. of Zoology

(8): 1-2.

Dezfuli, B.S. (1991): Histopathology in Leuciscus cephalus (Pisces: Cyprinidae)

resulting from infection with Pomphorhynchus laevis

(Acanthocephala). Parasitol., 33: 137-145.

Dike, S.C. and Read, C.P. (1971): Relation of tegumentory phosphohydrolase

and sugar transport in Hymenolepis diminuta. Journals of


Dollfus, R.P.H. (1934): Sur un cestode Pseudophyllidae parasite de Poisson d’

Ornement., Boll. Soc. Zool. France. 59 : 476-490.

Dollfus, R.P.H. (1937): Les Trematodes Digenea des selacieus (Plagiostomes)

catalogue par hotes. Distribution geographique. Ann. Parasit. 15:

259-281.

Dubinina, M.N. (1980): Tapeworms (cestode: Loguliidae) of the fauna of the

USSR. Amerind publishing co. pvt. Ltd, New Delhi, p. 320.

Dubois, G. and Pearson, J. (1963): Les Streigedia (Trematoda) de l’ Egypt. Ann

Parasitol 38: 91-92.

Dwivedi, M.P. (1965): On a new species of the Genus Genarchopsis Ozaki,

1925, from the stomach of Bufo melanostictus Schneid

(Hemiuridae, Trematoda). India Journal of Helminthology. Vol,

XVII No.1: pp. 37-42.

Eguchi, S. (1931): Studies on some parasites of Onchorynchus in Japan I. A new

trematode from Oncorhynchus macrostomus or “amago”. Osaka-

Koto. Igakukai Zassi: 6pp.

Ejswmont, L. (1931): Über die identität von Proshystera rosiltensis korkhaus

and Tanaisia fedtchenkoi Skrjabin, nebst einigen Bernerkungen

über Trematoden mit verbundenen Darmschenkeln. Ibidem (II):

531-547.

Ekman, E. and Norrgron, L. (2003): Pathology and immunohistochemistry in

three species of salmonids after experimental infection with

Flavobacterium psychrophilum. J. Fish Dis., 26: 529-538.

Erasmus, D.A. (1957): The phosphatase system of cestodes, part-2. Studies on

Cysticercus tenuicollis and Moniezia expansa (adult),

Parasitology. 47: 81-91.

Erasmus, D.A. (1957a): Studies on phosphatase systems of Cestodes. I. Studies

on Taenia pisiformis (cysticerus and adult). Parasitology 47: 70-

80.

Erasmus, D.A. (1957b): Studies on phosphatase systems of Cestodes. II. Studies

on Cysticerus tenuicollis and Moniezia expansa (adult).

Parasitology 47: 81-91.

Erasmus, D.A. (1972): The biology of trematodes. Edward Arnold, London.

Erasmus, D.A. and Ohman, C. (1963): The structure and function of the

adhesive organ in strigeid trematodes. Annals of New York

Academy of Science, 113, 7-35.

Ernest H. Williams, JR. and Williams G. Dyer (1993): Some digenea from

freshwater fishes of Alabama and Florida including Allocreadium

(Neoallocreadium) lucyae n.sp. (Digenea: Allocreadiidae). J.

Helmithol. Soc. Wash. 59 (1): pp. 111-116.

Esselman, E.J., Jianqiang, L., Crawford, D.J., Windus, J.L., Wolfe, A.D.

(1999): Clonal diversity in the rare Calamagristis porteri ssp.

Inserata (Poaceae): Comparative results for allozymes and random

amplified polymorphic DNA (RAPDs) and intersimple repeats

(ISSR) markers. Mol Ecol 8: 443-451.

Ewa Dzika, Tadewez Rolkiewiez and Rudolf Walter Hoffmann (2005):

Histopathological examination of liver and spleen of Roach,

Ratilus ratilus (L.). Originating from selected lakes of Warmia and

Mazury Labeland in Poland.

Fang, D.Q. and Roose, M.L. (1997): Identification of closely relate Citrus

cultivars with inter-simple sequence repeat markers. Theor Appl

Genet 95: 408-417.

Ferguson, H. W. (1989): Systemic pathology of fishes. Iowa State University

Press, Ames.

Fernando, C.H. and Furtado, J.I. (1964): Helminth parasites of some Malavan

freshwater fishes. Bulletin of the National Museum, State of

Singapore, 32: 45-71.

Fischthal, J.H. (1952): A redescription of Phyllodistomum lysteri miller, (19400

(Trematoda: Gorgoderidae) from the common white suckers. The

Jour. Parasit., Vol. 38, No.1. pp. 242-244.

Foresek, Z. and Rukavina, J. (1959): [Experimental immunization of dogs

against I Echinococcus granulossus. I. First observation.]

Veterinaria, Saraj., 8: 479-482.

Fujino, T., Kukuda, K., Hamajima, F. and Ishil, Y. (1989): Studies on host

specificity in Paraginimus westermani II. Histochemical and

cytochemical characterization of metacercariae and worms from

rats and dogs. J. Helminthol., 63 (4): 315-327.

Fujita, T. (1918): On a new species of Azygia. Japan J. Zool. Tokyo, 30: 269-

74.

Furtado, J.E. and Chaulan, L. (1971): Two new helminth species from the fish

Channa micropeltes, Cuvier (Ophicephalae) of Malaysia. Folia

Parasit, Praba 18 (4): 365.

Gasser, R.B., Woods, W.G., Blotkamp, C., Verweij, J., Storey, P.A. and

Poldermann, A.M. (1999): Screening for nucleotide variations in

ribosomal DNA Arrays of Oesophagustomum bifurcum by

polymerase chain reaction –coupled single-strand conformation

polymorphism. Electrophoresis 20: 1486-1491.

Gill, P., Jeffry’s. A.J. and Warrett, D.J. (1985): Forensic application of DNA

‘fingerprints’. Nature (Lond.) 318: 577-579.

Gittenberger, E., Groenenberg, D.S.J., Kokshoorn, B., Preece, RC. (2006):

Biogeography: molecular trails from hitch-hiking snails. Nature

409: 439-409.

Goeze, J.A.E. (1782): Versuch einer Naturgeschichte der Eing eweidewürmen

thierischer Korper. Xi + o471 pp. Blankenburg.

Goldberger, J. (1911): Some known and three new endoparasitic trematodes

from American freshwater fish. Bull. Hyg. Lab., U.S. Pub. Hlth &

Mar-Hosp. sew. 71: 7-35.

Gomori’s, G. (1946): Amer. J. Clin. Path; 16, Tech, Sect. 7, 177.

Gomori’s, G. (1950): Stain Tech., 25, 81.

Gopal Krishnan, V. (1968): Diseases and parasite of fishes in warm water

ponds in Asia and the far East, fisheries. Report. FAO-UN 445:

314-343. (Proceedings of the photo world symposium on warm

water pond fish culture).

Gozalez-Solis, D. Moravek, F. and Vidal-Martinez, M. (2002):

Procamallanus (Spirocamallanus chetualensis n.sp. Nematoda:

camallanidae) from the Mayan sea catfish, Ariopsis assimilis, off

Caribbean Coast of Mexico. J. Parasitol., 88: 765-768.

Gupta and Agarwal, S.M. (1984): Host-parasite relationships in Channa

punctatus and Euclinostomum laterostomum III. Transminase and

total proteins and free amino acids. Current Science, 53: 710-711.

Gupta S.P. (1951): On a new trematode, Phyllodistomum singhai n.sp. of the

family Gorgoderidae Looss, 1899 from the intestine of a

freshwater fish, Mastacembelus armatus (Lecepede, 1800). Indian

Journal of Helminthology, vol. III., No.1: 21-28.

Gupta, S. P. (1951): Three new trematodes of the family Hemiuridae Lucee.

1901 from freshwater fishes of U.P. Ind. Jour. helminth, Vol.III,

No. 1: 37-42.

Gupta, S.P. (1950): On a new trematode Allocreadium thapari n.sp. of the sub-

family Allocreadiidae Looss, 1899 from the intestine of a

freshwater fish Rita-rita (Ham.). Ind. J. Helm. 2 (1): 17-82.

Gupta, S.P. (1951 b): Studies on the trematode parasites of food – fishes of the

U.P. A new trematode, Macrotrema macroni n. gen., n.sp., from

the intestine of a freshwater fish, Macrones cavasius (Ham.) of the

sub family Leptophallinae Dayal, 1938. Ibid, 3: 101-108.

Gupta, S.P. (1953): Trematode parasites of freshwater fishes. Indian J.

Helminth. 5 (1): 1-80.

Gupta, S.P. (1956): two new trematode (Family: Allocreadiidae) from the

freshwater fishes of U.P. Ind. J. Helm. 8: 100-106.

Gupta, S.P. (1963): Two new trematodes of the family (Allocreadiidae Stossich,

1903), from the intestine of fresh water fishes of Banaras, U.P.

Proc. Helm. Soc. Wash. 30: 69-100.

Gupta, S. P. and Chakrabarti K.K. (1966): On a trematode Genarchopsis

thapari sp. from the intestine of snake from Lucknow, Ind. 18 (2):

177-180.

Gupta, S.P. and Gupta, R.C. (1977): Phosphatase activity in Geneo tigrinum

from Rana tigrina. Zenschrift fǜr Parasitenkunde, 54: 89-94.

Gupta, S.P. and Verma, S.L. (1976 Publ. 1977): On some trematode parasites

of freshwater fishes. Riv. Parasit. 37 (2/3): 171-182.

Gupta, V and Agarwal, S.K. (1979): Phosphatase activity in Gastrodiscus

crumenta (trematoda). Indian Journals of Parasitology, 3: 53-55.

Hafeez Md. (2001): Helminth parasites of public health importance- cestodes.

Journal of Parasitic Diseases, Vol. 25: 2-7.

Hayunga, E.G. (1977): Comparative histology of the scolices of three

caryophyllaeid tapeworms: Relation to pathology and site selection

in host intestine. Diss. Abs. Int., 38.

Hine, P.M. and Kennedy, C.R. (1974): Observation on the distribution

specificity and pathogenicity of the acanthocephalan

Pomphorhynchus laevis. J. Fish Biol., 6: 521-533.

Hiware, C.J. (1999): On a new tapeworm Senga armatusae n.sp. from

freshwater fish, Mastacembelus armatus at Pune (M.S.). Rivista Di

Parasit. XVI (LX), No. 1: 9-12.

Hiware, C.J. and Garad, V.K. (2002): Internal histopathology of Clarias

batrachus (Linn.) parasitized by Caryophyllaeid cestode. Inland

Fish Soc. India. 34 (2): 30-35.

Holes and Bethel, W.M. (1972): Modification of intermediate host behaviour

by parasites. In behavioral aspect of parasite transmission E.U.

Canning and C.A. Wright (eds.) academic press, New York, P:

123-149.

Holes, J.C. (1979): Parasitic population and host community structure. In Host-

parasite interfaces. B.B.Nikol (ed.) Academic Press Inc. New York,

P. 27-46.

Huglund, J., Engstrom, A., Morrison, D.A. and Mattson. J.G. (2004):

Genetic diversity assessed by amplified fragment length

polymorphism analysis of the parasitic nematode Dictyocaulus

viviparous the lungworm of cattle. International Journal for

parasitology. 34: 475-485.

Hundsdoerfer, A.K., Wink, M. (2006): Incongruence of morphology and

genetic marker in Hyles tithymali (Lepidoptera: sphingidae) from

in Canary islands. J. Zool syst Evol Res: 599-514.

Intapan, P.M., Wongkham, C., Imtawil, K.J. (2005): Detection of

Paragonimus heterotremus eggs experimentally infected cats by a

polymerase chain reaction-based method. J Parasitol. 91: 195-8.

Jacob H. Fischthal (1942): Three new species of Phyllodistomum (Trematoda:

Gorgoderidae) from Michigan fishes. The Journal of Parasitology,

vol. 28, No.4: 269-275.

Jacob H. Fischthal and Thomas J.D. (1972): Digenetic trematodes of fish from

the Volta River drainage in Ghana prior to the construction of the

Volta Dam at Akosomba in many 1964. Journal of Helminthology,

Vol. XLVI , No.1: 91-106.

Jadhav, B.V., Gavane, A.B. and Sarwade, B.W. (1990): On new

pseudophyllidae cestodes from Mastacembelus armatus of

Daryapur (M.S.) India. Rivista di Parasitol. 7: 19-22.

Jadhav, B.V. and Shinde, G.B. (1976): Three new species of genus


from a freshwater fish from Maharashtra. Marath. Univ. J. Sci.

(Nat. Sci.), XV (Sci. 8): 269-272.

Jadhav, B.V. and Shinde, G.B. (1980): On a new species Senga

aurangabadensis from Mastacembelus armatus. Biosearch (4):

25-27.

Jadhav, B.V., Ghavane, A.B. and Jadhav, A.P. (1991): Two new

Pseudophyllidean cestode from Mastacembelus armatus at

Daryapur (M.S.) India. Rivista Di Parasit. Vol. VIII (1): 19-22.

Jadhav, B.V., Ghavane, A.B. and Sarwade, B.W. (1990): On new

Pseudophyllidae cestodes from Mastacembelus armatus of

Daryapur (M.S.), India. Rivista di Parasitol. 7: 19-22.

Jadhav, B.V., Bhure, D.B. and Padwal Nitin (2005): A survey of cestode

parasites of freshwater fishes from Pune and Ahmednagar District

(M.S.) India. Proc. Recent Trends in Parasitology 30th. 48-51.

Jagdeshwari Dayal, D. Sc. (1949): Trematode parasites of Indian fishes, Part-II.

Indian Journal of Helminthology, vol.I , No.2: 93-116.

Jaiswal and narayan G. (1971): Azygia morulii n.sp. (Trematoda) from a

freshwater, Channa (Ophiocephalus) marulius, India. Folia

Parasitologica (Praha) 18: 165-168.

Jaiswal, G.P. (1957): Studies on the trematode parasites of fishes and birds in

Hyderabad State. Zool. Jaheb. Abt. Syst. 85: 1-72.

Jeffrey’s, A.J., Wilson, V. and Thein, S.L. (1985): Hypervaria positive

identification of an in migration test-case causing DNA

fingerprints. Nature (Lond.) 317: 818-819.

Jirawat Tudkaew, Norasing Penprapai, Rudolf Hoffmann and Kidchakan

Supamattaya (2008): Gonapodasmius epinepheli observed in

cage cultured orange spotted grouper (Epinephelus coioides) in

Southern Thailand: geographical distribution of parasite and host

response. Songklenkarin. J. Sci. Technol. 30 (2): 147-157.

Johnston, S.J. (1912): ‘On some Queenland trematodes with anatomical

observations and description of new species and genera’. Quar.

Jour. Micros. Sci., 59.

Johri, G.N. (1956): A new cestode Senga lucknowensis from Mastacembelus

armatus Lecep. Current Science, Bangalore, 25 (6): 193-195.

Jorge do Costa Eiras, Maria Luiza Gaspar Goulart Dias, Gilbert Cezar

Pavanelli and Marian Haruko Machado (1999): Histological

studies on the effect of Clinostomum marginatum (Digenea,

Clinostomidae) in its second intermediate host Loricariichthys

platymetopan (Osteichthyes, Loricardiidae.) of upper Parana river,

Brazil. Acta Scientiarum 21 (2): 237-241.

K. Buchmann (1998): Histochemical characteristics of Gyrodactylus riavini

parasitizing the fins of Rainbow trout (Onchorhynchus mykiss).

Folia Parasitol. 45: 312-318.

K.H. Kwak and C.H. Kim (1997): Characteristics of alkaline and acid

phosphatase in Spirometra erinacei. Korean J. Parasitol. 34 (1):

69-77.

Kadam, S.S., Jadhav, B.V. and Shinde, G.B. (1981): On a new cestode Senga

paithanensis n.sp. (Cestoda: Ptychobothriidae) from

Mastacembelus armatus. Bio-research, 5 (1): 95-96.

Kakaji, V.L. (1969): Studies on helminth parasites of Indian fishes. Part II. On

some species of the genus Allocreadium Looss, (1900). Annals

Parasit. Helm. Comp. 44 (2): 131-146.

Kalse A.T. and Shinde, G.B. (1999): Two new species of genus

Circumoncobothrium Shinde, 1968 (Cestoda: Pseudophylidea

Carus, 1863) from a freshwater fish at Khandesh (M.S.). Rivita Di.

Parasitol., XVI (LX) N.3: 195-198.

Karanis, P. and Taraschewski, H. (1993): Host-parasite interface of

Caryophyllaeus laticeps (Eucestoda: Caryophyllidae) in three

species a fish. J. Fish Dis. 16: 371-379.

Kaw, B.L. (1950): Studies in Helminth Parasites of Kashmir. Part I. Trematoda.

Ind. J. Helm. 2 (2): 67-126.

Kelejian, A., L. A. Schinazi and C.W. Schwabe (1961): Host-parasite

relationship in echinococcosis. V. Histochemical observations on

Echinococcus granulossus. J. Parasitol. 47: 181-188.

Kenneth L. Tiekotter and James R. Coggius (1982): Redescription of

Phyllodistomum americanum Osborn, 1903 with a Discussion of

the Taxonomic status of Phyllodistomum coatneyi Meserve, 1943

and Phyllodistomum bufonis Frondsen, 1957 (Trematoda:

Gorgoderidae) Proc. Helminthol. Soc. Wash. 49(2): 189-195.

Khalil, L.F. (1961): On a new trematode Orientocreadium lazeri sp. nov., from

a freshwater fish. Clarias lazera, in the Sudan. J. Helm. 35: 259-

262.

Koval V.P. (1949): Material to the knowledge of the Allocreadium Looss. Biol.

sbor: kiek. Universiteta No.4: 99-103.

Kroon, D.B. (1944): Enztmologia, 11: 186.

Lakshma Reddy, B., Banerjee, G., Rajender, G., Bikshapthi V., Swamy, M.

and Rama Rao, N.J. (2006): Histopathological and histochemical

abnormalities induced by Euclinostomum heterstomum in the liver

of freshwater fish, Channa punctatus. J. Aqua. Biol., Vol. 21 (2):

263-267.

Leidy, J. (1851): Contributions of Helminthology. Proc. Acad. Nat. Sci. Philad.

5: 201-10.

Levinsen, G. M. R. (1881): Bidrag til kundskab om Gr¢nlands trematodfauna.

Overs. K. Danske Vidensk. Selsk. Forh. 1: 52-84.

Lewis, F.J. (1935): ‘The trematode genus Phyllodistomum Braun.’ Trans. Amer.

Micros. Soc., 54.

Li J., Liao X. (2003): The taxonomic status of Digramma (Pseudophyllidae:

Ligulidae) inferred from DNA sequences. J. parasitol. 89: 792-

799.

Linton, E. (1910): Helminth fauna of the Dry tootugas, II. Trematodes. Carneg.

Inst. Wash. Publ. No. 133: 98 pp.

Linstow, O. Von (1907): Zwei neue Distomem aus Lucioperca Sandra der

Wolga. Ezhegodnik Zool. Mus. Akad. Nauk. Patrograd. 12: 201-2

Lio-Po, G.D., Lim, L.H.S. (2002): Infectious diseases of warm water fish in

fresh water. 231-281. In: Woo P.T.K., Bruno D.W., Lim L.H.S.

(eds.). Diseases and disorders of finfish in cage culture. CABI

Publishing, Wallingford, UK.

Looss, A. (1894): Die Distomen unserer FIsche und Frösche. Biblioth. Zool.,

XIV : 1-293.

Looss, A. (1899): Weitere Beitrage zur kenntnis der Trematodenfauna

Aegyptens zigleich Versuch einer naturlichen Gliederung des

Genus Distomum Retzius Zool. Jahrb. Syst. 12: 512-784.

Looss, A. (1900): Nachträgliche Bemerkungen zuden Hameu der von mir

Vorgeschlagemen Distomidengattugen. Zool. Anz., (xxiii) : 601-

608.

Looss, A. (1901): Notizen zur helminthologie Aegytens. IV uber Trematoden

aus Schildkraten der aegyptischen küsten Ibidem 30 (15-160): 555-

569, 618-625.

Looss, A. (1902): “Über neue und bekannte Trematoden aus Seeschilldkroton.

Erorterungen zur systematic und Nomenclatur,” Zool. Jahrb.,

Jena., Syst., 16: 411-894.

Luhe, M. (1899): Zur Kenntnis einiger Distomen. Zool. Anz. 22: 524-529.

Luhe, M. (1909): Trematoda. In Die Sćisswasserfauna Deutschland. Jena. Heft.

13.

Luhe, M. (1901): Zwei neue Distomen aus indischen Anuren. Ctbl. Bakt. I. Abt.

30: 166-177.

Luque, C., Legal, L., Staudter, H., Gers, C., Wink, M. (2002): Brief report

ISSR (intersimple sequence repeats) as genetic markers in

Noctuids (Lepidoptera). Herediteras 136: 251-253.

Lynch, J.E. (1936): Phyllodistomum singulare n.sp. a trematode from the

urinary bladder of Dicamptoden ensatus (Esch scholtz) with notes

on related species. J. Par. 22 (1): 42-47.

M. Stettler, M. Siles-Lucas, E. Sarciron, P. Lawton, B. Gottstein and A.

Hemphill (2001): Echinococcus multicularis alkaline phosphatase

as a marker for metacestode damage induced by in vitro drug

treatrment with albendazole sulphoxide and albendazole sulfone.

Antimicrob, Agents chemother. 45 (8): 2256-2262.

MacCallum (1917): Some new forms of parasitic worms. Zoopatologica 1: 43-

75.

MacCallum, G.A. (1921): Studies in helminthology. Part I- Trematodes, Part 2-

Cestodes, Part 3- Nematodes. Zoopathologica, 1 (6): 137-284.

McCoy, KD., Boulinier, T., Tirard C., Michalakis, Y. (2003) : Host dependent

genetic structure of parasite populations : differencial dispersal of

seabird tick host races. Evolution. 57: 288-296.

Mackiewicz, J.C. (1972): Parasitological review Caryophylldea (cestoda). A

review Expt. Parasit, 31: 417-512.

Mackiewiez, J.S., Cosgrove, G.E. and Gude, W.D. (1972): Relation of

pathology to scolex morphology among Caryophyllid cestodes. Z.

Parasitenkunde. 39: 233-246.

Madhavi, R. (1971): Life history of Genarchopsis goppo Ozaki, 1925

(Trematoda: Hemiuridae) from the freshwater fish Channa

punctatus. I. Helm. 52: 257-259.

Madhavi, R. (1978): Life history of Allocreadium fasciatusi Kakaji, 1969

(Trematoda: Allocreadiidae) from the freshwater fish, Aplocheilus

melastigma McCelland. J. Helm. 52: 51-59.

Madhavi, R. (1980): Life history of Allocreadium handiai Pande, 1937

(Trematoda: Allocreadiidae) from the freshwater fish Channa

punctatus (Bloch.). Z. Parasitenkd. 63: 89-97.

Madhavi, R. (2003): Metazoan parasites in fishes. In: Aquaculture Medicine

(I.S. Bright Singh, S.S. Pai, R. Philip and A. Mohandas, eds.).

Cochin University of Science & Technology, Cochin. 64-68.

Madhavi, R. and H. Rao K. (1977): Anatomy of the female reproductive

system in Allocreadium ophicephali Srivastava, 1960 (Trematoda:

Allocreadiidae). In Experta Parasitologion Memoria. 247-253.

Majid, M.A. and Shinde, G.B. (1984): Two new species of the genus Senga

Dollfus, 1934 (Cestoda: Pseudophyllidae) from freshwater fishes

at Jagnnathpuri, Orrisa. Indian Journal of Parasitol. (1): 169-172.

Maki, J. and Yauagisawa, T. (1980): Acid phosphatase activity demonstrated

in the nematodes, Dirofilaria inveritis and Angiosrtongylus

canteniensis with special reference to the character and

distribution, Parasitol., 80: 23-39.

Maleewong, W., Intapan, P.M., Wongkam, C. (2003): Detection of

Ophiochorchis viverrini in experimentally infected bithynid snails

and cyprinoid fishes by a PCR-based method, J Parasitol, 126: 63-

7.

Maltagliati, F., Lai, T., Casu, M., Valdesalici, S., Castelli, A. (2006):

Identification of endangered Mediterranean cyprinodontiform fish

by means of DNA inter-simple sequence repeats (ISSRs). Bichem

Syst Ecol 34: 626-634.

Manter, H.W. (1926): Some North American fish trematodes. Illin. Biol.

Monogr. 10 (2): 1-138.

Maqbool and Wajiha, Alam, S. Nizami (1984): Histochemical and

histoenzymological studies on the metacercaria of Clinostomum

complanatum (Trematoda: Digenea). Helminthologia, 21 (1): 21-

31.

Marshall, W.S. & Gillbert, N.C. (1905): Three new trematodes found

principally in black bass. Zool. Jb. Syst. 22: 477-88.

Mayberry, L.F. and Tibbitts, F.D. (1972): Hymenolepis diminuta (Order:

Cyclophyllidea): Histochemical localization of glycogen, neutral

lipids and alkaline phosphatase in developing worms. Zeitschrift

für parasitenkunde, 38: 66-76.

McCoy, KD., Boulinier, T., Tirard C., Michalakis, Y . (2003): Host-dependent

genetic structure of parasite populations: differencial dispersal of

seabird tick host races. Evolution 57: 288-296.

McGarry, J.W., Oritz, J.E. Hodgkinson, Goreish, I. and Williams, D.J.L.

(2007): PCR- based differenciation of Fasciola species

(Trematoda: Fasciolidae), using primers based on RAPD-derived

sequences. Annals of Tropical Medicine and Parasitology, Vol.

101, No. 5: 415-421.

Mehra H.R. (1966): Revision of Allocreadioidea Nicoll, 1934. Part I. Families:

Opeoelidae Ozaki, 1925; Opistholebetidae Fukui, 1929;

Allocreadiidae Stossich, 1903; Bunoderidae Nicoll, 1914;

Acanthocolpidae Luhe, 1909 and Pleorchiidae Poche, 1925. M.K.

Dikshit et Dikshit Press Allahabad. 1-46.

Minchella, D.J. and Scott, M.E. (1991): Parasitism: A cryptic determinant of

animal community structure. Trends in ecology and evolution, 6:

250-254.

Misako Urabe (2001): Life cycle of Genarchopsis goppo (Trematoda:

Derogenodae) from Nara, Japan. J. Parasit. 87(6): 1405-1408.

Mitchell, A.J. (1982): Pathogenicity and histopathology of an unusually intense

infection of white grubs (Posthodiplostimum m. mininum) in the

fathed minnow (Pimephales promelas), Journal of wildlife

Diseased Vol. 18. No. 1: 51-57.

Mitra, K.B. and Shinde, G.B. (1980): Histopathology of cestode A. Indiana

(Cohn, 1900), Gallus domenticus, at Aurangabad, India. Curr. Sci.

Vol. 49 (5): 206-207.

Molnar, K. (2005): Histopathological changes caused by the metacestodes of

Neogryporhynchus cheilaneristrotus (Wedl, 1855) in the gut of the

Gibel carp, Carassius gibelio. Acta veterinaria Hungarica, 53 (1):

pp. 45-52.

Molnar, K., Majoros, g., Csaba, Gy. And Szekely, Cs. (2003): Pathology of

Atractilytocestus huronensis Anthony, 1958 (Cestoda,

Caryophyllidae) in Hungarian pond farmed common carp. Acta

Parasitol. 48: 222-228.

Mongkol-Primpol (2000): Study on diseases in Gunter’s walking catfish

(Clarias batrachus) and its prevention. Abstr. M.Sc. Theses fish.

Sci. (1985-1900. 2000). Kesetsart University, Bangkok. 33-34.

Monzer Hasnain (1992): On a new cestode Senga chauhani n.sp. from fish host,

Channa punctatus from Jamshedpur. National Journal of

Helminthology. Vol. XXXXIV No. 1: 123-127.

Mortens, E. and Moens, J. (1995): The metazoans ecto-and endo-parasites of

the rabbit fish, Siganus sutor (Cuvier & Valanciennces, 1835) off

the Kenya coast. Afr. J. Ecol. 33: 405-416.

Motwani, M.P. and Srivastava C.B. (1961): On two Phyllodistomes from the

urinary bladder of siluroid fishes (Trematoda: Gorgoderidae).

Indian Journal of Helm. Vol. XIII , No.2, pp. 93-99.

Muley, V.B. (1972): Study on the trematode parasites of vertebrates Thesis

submitted to Marahwada University, Aurangabad, (M.S.), India,

pp. 87-89.

Murai, E., Molnar, K., Gubanyki, A. (1997): Occurrence of the adult of

Paradilepis scolecina (Rudolphi, 1819) (Cestoda: Dilepididae) in

lake balaton, Hungary-Parasit. Hung., 29-30 (2): 33-38.

Murlidhar, A. and Shinde, G.B. (1987): Histopathology of cestode,

Acanthobothrium uncinatum (Rudoiphi, 1819) from Rhynchobatus

ajeddensis at Kakinada, A.P. India. Indian. J. of Parasitology

11(1): 85-86.

Muzzal Patrick M. (1980): Population biology and host-parasite relationships

of Tryganodistomum attenuatum (Trematoda: Lissorchidae)

infecting the white sucker, Catostomus commersoni (Lacepede). J.

Parasitol., 66(2): 293-298.

Nagaty, H. F. (1930): ‘A new Anaporrhutine trematode genus and species,

Nagmia yorki with a review of the classification of the sub-

family’. Ann. Trop. Med. Parasit., 24.

Nei, M. and Li, WH (1979): Mathematical model for studying genetic variation

in terms of restriction endonuclease. Proceeding of the National

Academy of Sciences of the United States of America. 76: 5269-

5273.

Nilima M. kankale (2008): A new species of the genus Senga nathsagarensis

from freshwater fish, Mastacembelus armatus. National Journal of

Life Sciences, 5 (3): 81-84.

Niyogi, A. and Agarwal, S. M. (1989): Intestinal pathology due to

caryophyllaeids parasitizing Clarias batrachus (Linn.) at Raipur.

Indian J. Helminth. 41: 112-119.

O. Poljakova-Krustena, Y.A. Mizinsha-Beha and S.A. Stoitsova (1983):

Cytochemical study of some phosphatases in the tegument of two

cestode species. Helminthologia 16: 64-67.

Odhner, T. (1901): Revision einiger Artender Distomen-galtung Allocreadium

Lss. Zool. Jarbs. Syst., XIV: 483-520.

Odhner. T. (1911): Zum natürlichen system der digenen Trematoden Ii. Familie

Zoogonidae n.fam. Zool. Anz. 37: 237-53.

Odhner, TH. (1902): Milteilungen Zur Kenntniss der Distomen. Centrambl. F.

Bakt. U. Parasit. XXXI: 152-162.

Odhner, TH. (1928): Weitere Trematoden mit Anus. Ark. Zool. 20 B (2): 1-6.

Oftenheim, E. von (1900): Über eine neue Distomidengahung Inang Dist. 42 pp.

Z. Naturg. 73: 145-186.

Ohman, C. (1966): Histochemical study of the enzyme activity of gland cells in

certain adult trematodes. Proceedings of the first international

congress of Parasitology. Romr 1964. 1: 77.

Oide, M. (1970): Purification and some properties of alkaline phosphatase from

intestinal mucosa of the eel adapted to fresh water or sea water.

Comp. Biochem. Physiol. 36: 241-252.

Okamoto, M., Nakao, M., Sako, Y., Ito, A. (2001): Molecular variation of

Teania solium in the world. Southest Asian J Trop Public Health:

32 (Suppl 2): 90-3.

Okamoto, M., Veda, H., Hayashi, M., Oku, Y., Kurosawa, Y. and Kamiya,

M. (1995): Application of DNA fingerprinting with

diagoxigenated oligonucleotide probe (CAC)5 to analysis of the

genetic variation within Taenia taeniaeformis. J. vet. Med. Sci. 57:

262-272.

Olsen PD, Tkach VV. (2005): Advances and trends in the molecular systematics

of the parasitic platyhelminthes. Adv. Parasitol 60: 165-243.

Otsen, M., Hoekstra, R., Plas, M.E., Buntjer, J.B., Lenstra, J.A. and Roos,

S.H. (2001): Amplified fragment length polymorphism analysis of

genetic diversity of haemonchus contortus during selection for

drug resistance. International Journal for parasitology 31: 1138-

1143.

Ozaki, Y. (1924): On a new species of trematode of the genus Azygia (Japanese

text). Dabuts. Zasshi, Tokyo, (432) 36: 420-35.

Ozaki, Y. (1925): On a new genus of fish trematode with anus. J. Par. 12: 51-

53. Zool. Mag. 37: 416-423.

Ozaki, Y. (1926): On two new trematodes from freshwater fishes of Japan. Zool.

Mag (Dobutsagaku Zassi) 38: 124-130.

Ozaki, Y. and Asada, J. (1926): A new human trematode, Hetrophyes

katsuradai n.sp. J. Par. 12 (4): 216-218. Arch. Schiffs-u.

Tropenhyg. 30 (9): 360-365.

P. Pal and V. Tandon (1998): An helminthic efficacy of flemingia nevestita

(Leguninoceae. Genistein-induced alteration in the activity of

tegumental enzymes in cestode, R. echinobithria. Parasitol. Int.,

47 (1): 233-243.

Pande, B.P. (1934): On a new trematode from an Indian freshwater fish. Proc.

Acad. Sci. U.P. Agra & Oudh, India 4: 107-112.

Pande B. P. (1937): Morphology and relationship of a new digenetic trematode

from an Indian fresh water fish. Ann. Mag. Nat. Hist. Ser. 10 (20):

415-421.

Pande, B.P. (1938): The trematode genus Allocreadium in North Indian fresh

water fishes. Proc. Ind. Acad. Sci. 7 (2): 54-60.

Pande, B.P. (1938 a): The trematode genus Allocreadium in North Indian fresh

water fishes. Proc. Ind. Acad. Sci. 7 (2): 54-60.

Pande, B.P. (1938 b): On two new trematodes from Indian cyprinoid fishes

with remarks on the genus Allocreadium Looss. Proc. Nat. Acad.

Ac. India. 8 (4): 110-115.

Pande, P.N., Mamta, T. and Neetu, M. (2006): On two new species of genus

Senga Dollfus, 1934 (Family: Ptychobothriidae) Luhe, 1902 from

the intestine of freshwater fishes. Indian Journal of

Helminthology. Vol. 24:

Paperna, I. and Vanas, J.G. (1983): The pathology of Childenella hexastica

infections in cichlid fishes. J. Fish Biol., 23 (4): 441-450.

Patil, D.N. and Jadhav, B.V. (2003): On a new species the genus Senga

Dollfus, 1934 (Cestoda: Ptychobothridae) Luhe, 1902 as Senga

tappi n.sp. from M. armatus from the Shripur dist. Dhule (M.S.). J.

Com. Tox. Phy. Vol. 1: 68-72.

Patil, S.R., Shinde, G.B. and Jadhav, B.V. (1998): A new species of the genus

Circumoncobothrium Shinde, 1968 (Cestoda: Pseudophyllidae)

Carus, 1863 from Mastacembelus armatus at Vadgaon, (M.S.)

India. Journal of Para. Diseases. 22 (2): 148-151.

Paul, C. Stromberg and John Crites (1974): Triaenophoriasis in Lake Erie

white bass, Macrone chrysops, Journal of wildlife Diseases Vol.

10: 352-358.

Pawar, S.B. (2002): A new species Circumoncobothrium armatusae n.sp.

(Cestoda: Pseudophyllidae) from Mastacembelus armatus at

Paithan, India. Riv. Di. Parasit. Vol. XX (LXIII) No.3: 219-222.

Pearse, A.S. (1920): The fishes of Lake Valencia, Venezuela. Univ. Wisc. Stud.

In Sci. 1: 51.

Pearse, A.S. (1924): Observations on parasitic worms from Wisconsin fishes.

Trans. Wis. Acad. Sci. Arts Lett. 21: 147-160.

Pennott-De Cooman, E. and G. van Grebergen (1947): Aanvullend onderzock

over de phosphatasen bij de plathelminthen. Natuurwet. Tijdeschr.

(Belg.) 29: 9-12.

Pheravut Wongsawad and Chalobol Wongsawad (2007): DNA fingerprints of

some heterophyid trematodes from adult and metacercarial stages

in Thailand. Southeast Asian J. Troup Med Public Health, Vol. 38

(suppl I): 110-114.

Pippy, J.H.C. (1969): Pomphorhynchus laevis in Atlantic salmon and its use as

a biological log. J. Fish Res. Can., 26: 909-919.

Poulin, R. and Morands, S. (2000): The diversity of parasites. Q. Rev Biol 75:

277-273.

Poynter, D. (1966): Some tissue reactions to the nematode parasite of animals.

In: Advances in parasitology (Ben Daws, ed.). Academic press.

London a & New York, 15: 321-328.

Probert, A.J. and Lwin, T. (1974): Kinetic properties and location of non-

specific phosphomonoesterases in subcellular fractions of Fasciola

hepatica. Experimental Parasitology, 35: 253-261.

R.H. Fetterer and M.L. Rhoads (2000): Characterization of acid phosphatase

and phosphorylcholine hydrolase in adult Haeminchus confortus.

J. Parasitol. 86 (1): 1-6.

Radhakrishana, S., Nair, N.B. and Balasubramanian, N.K. (1983): Adult

cestode infection of the marine teleost fish, Saurida tumbil

(Bloch.), Acta icthyologiia et Piscatoria Vol. XIII Fasc. 1: 75-95.

Rai, P. (1970): A redescription of Allocreadium mehrai Gupta, 1956 and A.

handiai Pande, 1937 with a key to identification of species of

Allocreadium Looss, 1900 from freshwater fishes of India. Ind. J.

Sci. Indust. (B) 4 (1): 35-40.

Rai, S.L. (1962): Studies on three new species of the genus Allocreadium Looss,

1900 from the intestine of Barbus tor (Ham.) Parasit. 54: 23-30.

Rai, S.L. (1964): Azygia stunkardi sp. Nov. (Trematoda: Azygiidae).


Ramadevi, P. and Rao, K.H. (1966): Plerocercoid of Senga n.sp.

(Pseudophyllidea, ptychobothriidae) from ferahwater fish,

Panchax panchax (Ham. and Buch). Current Sci. 35 (247): 626-

627.

Ramadevi, P. and Rao (1973): On Senga visakhapatnamensis n.sp. (Cestoda:

Pseudophyllidea) from the intestine of the freshwater fish

Ophiocephalus punctatus (Bloch.). Rivista Di Parasitol., Vol. 34,

No. 4: 281-286.

Raymond, C. Bowen (1969): Histology and histochemistry of the capsule of

Protocephalus sp. (cestoda) in Lepomis macrochirus. The Ohio

Journal of Science, 69 (4): 243.

Read, C.P. (1966): Nutrition of intestinal helminths. In: Biology of parasites,

E.J.L. Soulsby, ed. New York: Academic press.

Reddy B. Lakshma, G. Banerjee, G. Rajender, V. Bikshapthi, M. Swamy

and N.J. Rama Rao (2006): Histopathology and histochemical

abnormalities induced by Euclinostomum heterostomum in the

liver of freshwater fish, Channa punctatus. J. Aqua. Biol., Vol. 21

(2): 263-267.

Ress, G. (1967): Pathogenicity of adult cestodes. Helminthological Abstract, 36:

1-23.

Revenga, J.E., Torries, P. and Siegmund, I. (2006): Galaxias maculates

(Galaxiidae, Salmoniformes) infected with Acanthostomoides

apophalliformis (Digenea, Platyhelminthes) in Southern

Argentina. Pathology and absence of parasite induced mortality.

Braz. J. Vet. Res. Anim. Sci., Sao Paulo, V. 43 N.5: p. 642-646.

Robert, E. Olsen and Jack, R. Plerce (1997): A trematode metacercaria

causing gill cartilage proliferation in steelhead Trout from Oregon.

Journal of wildlife Diseases, 33(4): 886-890.

Robert, R.J. (2001): Fish Pathology. 3rd Edn. W.B. Saunders, London &

Phildelphia.

Robinson, E.M. and Williams, T.C. (1971): Relationship of some species of

digenea with the marine prosobranch Littorina littorina (L) III.

The effect of larval digenea on the glycogen content of the

digestive gland of L. littoria, J. Helm., 45 (4): 381-401.

Robinson, W.A., Liston, A., Doescher, P.S., Svejcar. (1997): ISSR markers

quantity clonal sexual reproduction in Fesruca idahonensis

(Poaceae). Am J Bot 84: 89.

Rogers, W.P. (1947): Histochemical demonstration of phosphatases in Moniezia

and Ascaris nature 159:347.

Rottman, R.W., Francis-Floyd, R. and Durborow, R. (1992): The role of

stress in fish disease. Southern Regional Aquaculture Center

Publications. 474.

Roy, T.K. (1979): Histochemical studies on Raillietina (Raillietina) johri

(Cestoda: Cavaineidae). I. Nonspecific and specific phosphatases.

Journal of Helminthology, 53: 45-49.

Rudolphi, C.A. (1802): Fortsetzung der Beobachtungen ǜber die

Cingeweidewǜemen. Arch. Zool. U. Zoot. 2 (3): 61-125.

Rudolphi, C.A. (1808): Entozoorum sive vermium intestinalium historia

naturalis. Vol. 1. Amstelaed. 527.

Rudolphi, C.A. (1819): Entozoorum synopsis, Berol., 811.

Ruhela, S., Pandey, A.K. and Khare, A.K. (2006): Effect of experimental

Procamallanus infection on certain blood parameters of the

freshwater catfish, Clarias batrachus (Linnaeus). J. Ecophisiol.

Occup. Hlth., 6: 73-76.

Saksena, J.N. (1958): Studies on two new species of the genus Orientocreadium

(Trematoda: Allocreadiidae) from the intestine of Clarias magur.

Proc. Nat. Acad. Sci. India, sec. B 28: 58-64 (Issues 8 Aug.).

Saksena, J.N. (1960): Studies on a new species of the genus Orientocreadium

(Trematoda: Allocreadiidae) from the intestine of Clarias magur.

Ibid. 30: 83-86.

Satpute, L.R. and Agarwal, S.M. (1974): A “Diverticulosis” of the fish

duodenum infected with cestodes. Indian J. Exper. Biol., 12: 373-

375.

Satpute, L.R. and Agarwal, S.M. (1974 a): Seasonal infection of Clarias

batrachus (Bloch) by Lytocestus indicus Moghe and parasitic

effects on its haematology and histopathology. Indian J. Exp.

Biol., 12: 584-586.

Schaperclaus, W. (1954): Fischkrankheitan. Academie-verlag, Berlin, 708 p.

[cited by Wales, J.H. 1958. Calif Fish and Game, 44: 125-136]-

243-248.

Schnurrenberger, P. (1975): Diseases transmitted from animals to man.

Sprinffield, Illinois, USA 646-677.

Sharma, P.N. (1976): Histochemical study on the distribution of alkaline

phosphatase, 5-nucleotides and ATPase in various reproductive

tissues of certain digenetic trematodes. Zeitschrift Fǜr

Parasitenkunde, 49: 223-231.

Sharma, P.N. and Hanna, R.F. (1988): Ultrastructure and cytochemistry of the

tegument of Orthocoelium scoliocoelium and Paramphistomum

cervi (Trematode: Digenea). J. Helminthol., 62 (4): 331-343.

Shiff, C., Broumer, K.C., Clow, L. (2000): Schistosoma haematobium

population genetics of Schistosoma haematobium by direct

measurement of parasite diversity using RAPD-PCR. J. Exp

Parasitol. 96: 47-51.

Shimazu, T. (1988): Trematodes of the genus Allocreadium (Allocreadiidae)

from freshwater fishes of Japan. Bulletin of the National Science

Museum, Tokyo, Ser. A, 14: 1-21.

Shinde, G.B. (1968): On Circumoncobothrium ophiocephali n. gen. n.sp. from

freshwater fish, Ophiocephalus leucopunctatus in India, Rivista Di

Parasitol. 19 (20): 111-114.

Shinde, G.B. (1972): Studies on Indian cestode redescription of Senga benesardi

Dollfus, 1923, Marathwada Univ. J. Sci. 11: 39-40.

Shinde, G.B. (1976): On a new species of Circumoncobothrium Shinde, 1968

(Cestoda: Pseudophyllidea Carus, 1863) from fresh water fish,

M.S. Ibid., XVI : 129-133.

Shinde, G.B. and Jadhav, B.V. (1980): New tapeworm, Senga godavari from

Mastacembelus armatus, Aurangabad (M.S.) India. Biology. Vol.

II 40: 46-48.

Shinde, G.B. and Kalse, A.T. (1999): Two new species of genus

Circumoncobothrium Shinde, 1968 (Cestoda: Pseudophyllidae)

Carus, 1863 from fresh water fish at Khandesh (M.S.), Rivista Di

Parasitol., (IX), No. 3: 209-215.

Shinde, G.B., Pawar, S.B. and Chauhan, S.P. (2002): A new species

Circumoncobothrium mastacembellusae n.sp. (Cestoda:

Pseudophyllidae) from Mastacembelus armatus at Paithan, India.

Riv. Di. Parasit., Vol. XX (LXII) No. 3: 195-198.

Shinde, G.B., Sarwade, D.V., Jadhav, B.V. and Mahagan, M.A. (1994): On a

new species of the genus Circumoncobothrium Shinde, 1968

(Cestoda: Pseudophyllidae) Carus, 1863 from Mastacembelus

armatus (Cuv. and Val.) from freshwater fish at Aurangabad

(M.S.) India. Rivista Di Parasitologia 11 (55): 167-169.

Simmons, M.P., Zhang, L.B., Webb, C.T., Muller, K. (2007): A penalty of

using anonymous dominant markera (AFLPs, ISSR and RAPDs)

for phylogenetic inference. Mol Phylogenet Evol 42: 528-542.

Singh, R.N., Swarnjeet and Davies, R.W. (1984): Ectoparasites of the

freshwater food fishes of Haryana (India). Proc. Indian Acad. Sci.

Anim. Sci. 93 (7): 663-670.

Sinha, K.P. (1988): Procamallanus (Camallanidae: Nematoda) infection in the

fish, Clarias batrachus. Environ. And Ecol., 6: 1035-1037.

Sinha, K.P. (2000): Haematalogical manifestation in Clarias batrachus carrying

helminth infections. J. Parasit. Dis., 24: 167-170.

Smyth, J. D. (1969): Parasites as biological model. Parasitol., 59: 73-92.

Southwell, T. (1913): Parasites of fishes-notes of Bengal fisheries laboratory.

Rec. Ind. Mus. 9: 98-99.

Sripalwit, P., Wongsawad, C., Chai, J.Y., Rojanapaibul, A.,

Anantalabhochai, S. (2003): Development of HAT-RAPD

technique for the identification of Stellantchasmus faleatus

[Abstract.] Bangkok: 4th seminar on food – and Water-borne

parasitic ziinoses, 2nd International meeting on Gnathostomiasis

and joint International Tropical Medicine Meeting, 289.

Srivastava, C.B. (1975): Fish pathological studied in India. A brief Review Dr.

B.S. Chauhan Comm. Vol. pp. 349-358.

Srivastava, H.D. (1933): On new trematodes of frog and fishes of the United

Provinces, India. Part I. Diatoms of the family Hemiuridae from

North Indian fishes and frogs with a systematic discussion on the

family Halipegidae and the genera Vitellotrema Guberlet and

Genarchopsis Ozaki. Bull. Acad. Sc. U.P. Agra and Oudh.

Allahabad 3 (1): 41-60.

Srivastava, M. and Gupta, S.P. (1975): Phosphatase activity in Isopaporchis

hypselobagii (trematoda). Zoologischer Auzeiger 195: 89-94.

Srivastava, P.S. (1960): On a new trematode Allocreadium ophiocephali n.sp. of

the family Allocreadiidae Stossich, 1904 from the intestine of

Ophiocephalus punctatus. Indian Journal of Halminthology, Vol.

XII, No. 2: 108-113.

Stafford, J. (1904): Trematodes from Canadian fishes. Zool. Anz. 27: 481-95.

Steelman, M. Gerald (1938): A description of phyllodistomum caudatum n.sp.

The American Midland naturalist. Vol. 19, No. 3: 613-616.

Stephenson, L. S., Pond, W.G., Nitsheins, M.C., Krook, L.P. and Crompton,

D.W.T. (1980): Ascaris suum: Nutrient absorption growth and

intestinal pathology in young pigs experimentally infected with 15

days old larvae. Exp. Parasitol., 49: 15-25.

Stossich, M. (1885): Brani di elmintologica tergestina. Boll. Soc. Adriat. Sc. Nat.

Trieate, Series 2. ix: 1-9.

Stossich, M. (1900): Osservationi elmintologiche. Bull. Soc. Adriat. Sc. Nat.

Trieste. XX : 89-103.

Stossich, M. (1903): Una nuova specie di Helicometra Odhner. Arch. Parasit.,

Vii, 373-376.

Stossich, M. (1904): Note distomologiche. Boll. Soc. Adriat. Sc. Nat. Trieste,

xxi, 193-201.

Stossich, M. (1905): Note distomatologiche 3. Bull. Soc. Adriat. Sc. Nat. 22:

211-227.

Stunkard, H.W. (1956): The morphology and life cycle of digenetic trematodes,

Azygia Sebago Ward, 1910, Biol. Bull., Woods Hole, 111: 248-68.

Supugade, et al., (2005): Circumoncobothrium vitellariensis n.sp.

Ptycobothriidae (Luhe, 1920) from Mastacembelus armatus

(M.S.), India. Trajectory. Vol. 13 No. 1: 43-49.

Suzana B. Amato and J.F.R. Amato (1993): A new species of Phyllodistomum

Braun, 1899 (Digenea: Gorgideridae) from Rhamdia Queen (Quoy

and Gaimard, 1824) (Siluriformes: Pimelodidae). Mem. Inst

Oswalto Cruz, Rio de Janeiro, vol.88 (4): 557-559.

Symons, L.E.A. and Fairbairn, D. (1963): Comparative efficacy of

serodiagnostic test in buffaloes naturally infected with Fasciola

gigantica. Ind. J. Amer. Sci., 57: 373-376.

Takeshi Shimazu (1990): Adult digenetic trematodes parasitic in freshwater

fishes of Hakkaido, Japan. A review. Sci. Rep. Hakkaido Fish

Hatchery, 48: 69-78.

Takeshi Shimazu (2003): Two new species of the Genus Allocreadium

(Digenea: Allocreadiidae) from a freshwater fish in Nagano,

Central Japan. Bull. Natn. Sci. Mus. Tokyo, Ser. A, 29 (3): pp. 119-

123.

Takeshi Shimazu, Shigchiko Urawa and Claudio O. Coria (2000): Four

species of digeneans including Allocreadium patagonicum n.sp.

(Allocreadiidae), from freshwater fishes of Patagonia Argentina.

Folia Parasitologia 47: 111-117.

Tandon, R.S. and Misra, K.C. (1978): Acid and alkaline phosphatase activities

in Fasciolopsis buski (Lankester, 1857) Odhner 1902. Indian

Journal of Parasitology, 2: 145-146.

Tat, M.B. and Jadhav, B.V. (1997): Senga mohekare n.sp. (Cestoda:

Ptychobothriidae) from Mastacembelus armatus at Pune (M.S.).

Riv. Di Parasit. Vol. XVII (LVIII), No. 2: 203-296.

Tat, M.B. and Jadhav, B.V. (2004): A new species of the genus


Carus, 1863 from Ophiocephalus gachua at Dhanegaon District,

Beed. Nat. Jour. of Life Sciences. 1 (1): 129-132.

Tadros, G. (1968): A redescription of Polynchobothrium Clarias (Woodland,

1925) Meggit, 1930 (Cestoda: Bothricephallidae) with a brief

review of the genus, Polyonchobothrium. Diesing, 1854 and the

identify of the genera Teracampos Wedl, 1961, Senga Dollfus,

1934 and Onchobothiocephalus Yamaguti, 1959.

Thorsell, W. and Bjorkman, N. (1965): Morphological and biological studies

on absorption and secretion in alimentary tract of Fasciola

hepatica L . Journal of Parasitology, 51, 217-223.

Threadgold, L.T. (1968): Electron microscope studies of Fasciola hepatica. VI .

The ultra structural localization of phosphatases. Exp. Parasit. 13,

264-276.

Travassos, L. (1920): “Contribuicoes para O conhecimento da fauna

helminthological brasileira. XIV. Especies brasilerias da familia

gorgoderidae Looss, (1901)”. Braz. Med. Ann., 36: 17-20.

Travassos, LO. (1926): Trematodes intestinaes dos batrachios do Brazil.

(Addenda). Sc. Med. 4 (1): 89.

Teseng (1933): Study on some cestodes from fishes. Jour. of Science National

Univ. Shantuma, Tsingtao, China (2): 1-21.

Tsin. (1933): Azygia hwangtsinyii. In Husti & Li. Chinese med. J. (1940), p. 559.

Tubangi, M.A. (1928): Trematode parasites of Philippine vertebrates. Philipp. J.

Sci. 36: 251-71.

Tubangi, M.A. (1931): Trematode parasites of Philippine vertebrates, III: Flukes

from fish and reptiles. Phil. J. Sci. 54: 415-423.

Tubangi, M.A. (1933): Trematode parasites of Philippine vertebrates, IV.

Description of new species and classification. Ibid. 52: 167-197.

Valtoneu, E. T., Haaparanta, A., Hoffmann, R. (1994): Occurrence and

histological response of Raphidascaris acus (Nematoda-

Ascaridoidea) in roach from four lakes differing in water quality.

Int. J. Parasit., 24: 197-206.

Velasquez, C.C. (1958): Notes on Azygia and related genera (Digenea:

Azygiidae). Proc. Helminth. Soc. Wash. 25: 91-4.

Vladimir V. Besprozvamykh, Alexey V. Ermolenko and Marty R. Deveney

(2009): Orientocreadium elegans n.sp. and Orientocreadium

pseudobergi Yamaguti (Digenea: Orientocreadiidae) from fresh

water fish of the Primorsky region (Southern far east, Russia)

withy a description of their life cycles. Zootaxa 2176: 22-32.

Waitz, J.A. (1963): Histochemical studies of the cestode, Hydatigera

taeniaeformis Batsch, 1786. J. Parasit. 49: 73-80.

Waitz, J.A. and Schardein (1964): Histochemical studies of four

cyclophyllidean cestodes. J. Parasit. 50: 271-277.

Waldschmidt A.M., Salomão T.M.F., Barros E.G., Campos L.A.O. (1997)

Extraction of genomic DNA from Melipona quadrifasciata

(Hymenoptera: Apidae, Meliponinae), Genet. Mol. Biol. 20: 421-

423.

Wallace A. Evans (1974): The histopathology of cutthroat trout experimentally

infected with the blood fluke Sanguinicola klamathensis. Journal

of Wildlife Diseases, Vol. 10: 243-248.

Wallin E. Ivan (1909): A new species of the trematode Genus Allocreadium

with a revision of the genus and a key to the sub-family

Allocreadiinae. Transactions of the American microscipial Society.

Vol. 29: No. 1, 50-66.

Wang Xiyum (1984): Parasitic trematodes from Boyang Lake fishes V.

Gorgoderidae Looss, 1901 with description of two new species.

Oceanologia et Limnologia Sinica. Vol. 15, No. 1: 98-104.

Wanstali, S.T., Thomas, J.S. and Robotham P.W.J. (1986): the pathology

caused by Pomphorynchus laevis mǜller (Acanthocephalus) in the

alimentary canal of the stone loach, Nemacheilus barbatulus (L.).

J. Fish Dis., 11: 511-523.

Ward, H.B. (1910): Internal parasites the Sebago salmon. Bull. Bureau Fish. 28:

1151-94.

Wardle, R.A.Mcloed, J.A. and Radinoky, S (1974): Advances in the Zoology

of tapeworms, 1950-1970. Univ. Minnesota. Minnetoba Press,

Monneapolis, 1-274.

Williams, H.H. (1967): Helminth disease if fish. Helminthol. Abstr., 36: 261-

295.

Wolfe, A.D., Xiang, Q.Y., Kephart, S.R., (1998a): Assesssing hybridization in

natural populations of Peustemon (Scrophulariaceae) using

hypervariable intersimple sequence repeat (ISSR) bands. Mol Ecol.

7: 1107-1125.

Wolfe, A.D., Xiang, Q.Y., Kephart, S.R., (1998b): Diploid hybrid speciation in

Pentemon (Scrophulariaceae). Proc. Natl. trend. Sci. USA 95:

5112-5115.

Wongratanachewin, S., Pumidonming, W., Sermswan, R.W., Maleewong,

W. (2001): Development of a PCR-based method for the detection

of Opisthorchis viverrini in experimentally infected hamsters. J.

Parasitol. 122: 175-180.

Wongsawad, C., Tanu Marayang, Jadhav, B.V. (1998): Circumoncobothrium

baimaii n.sp. (Cestoda: Pseudophyllidae) from fresh water fish,

Maesa stream Chiang Mai, Thailand. Rivista Di Parasitologia.

Vol. XV (LIX) No. 3: 291-294.

Woodland, W.N.F. (1934): On a new Bothriocephalus and a new genus

Bothriocephalidae from Indian freshwater fishes. Parasit. 16: 441-

451.

Wu, W., Zheng, Y.L., Chen, L., Yang, R.W., Yan, Z.H. (2005): Evaluation of

genetic relationship in the genus Houttuynia Thumb in China

based on RAPD and ISSR markers. Biochem syst Ecol 33: 1141-

1157.

Yamaguti, S. (1934): Studies on the helminth fauna of Japan. Part 2. Trematodes

of fishes, I. Jap. J. Zool. 5: 249-541.

Yamaguti, S. (1937): A new trematode from Amyda japonica. Jap. J. Zool.,

7(30: 505-506.

Yamaguti, S. (1953): Idem 3. Digenetic trematodes of fishes. 11, Ibid. 8 (3):

257-285.

Yamaguti, S. (1956): Systema Helminthum Vol-II. The cestode of vertebrates.

Interscience publ. New York and London, 1-860.

Yamaguti, S. (1958): Systema Helminthum Vol. 1. The digenetic trematodes of

the vertebrates. Interscience. New York. 1575pp.

Yamaguti, S. (1971): A synopsis of digenetic trematodes of vertebrates vol-

I&II. Keigaku Publishing Co. Ltd. Japan.

Yamao, Y. (1952): Histochemical studies on endoparasites. VII. Distribution of

the glyceromonophosphatases in the tissues of the cestodes

Anoplocephala perfoliata, A. magna, Moniezia benedeni, M.

expansa and Taenia taeniaeformis. Zool. Mag. (Japan) 61: 254-

260.

Yu JR, Chung JS, Huh S, Lee SH, Chai JY. (1997b): PCR-RFLP patterns of

three kinds of Metagonimus in Korea. Korean J. Parasitol. 35:

271-6.

Yu JR, Guk SM, Han ET, Chai JY. (1997a): Molecular differentiation of three

species of Metagonimus by simple sequence repeat anchored PCR

(SSR-PCR) amplification. J. Parasitol. 86: 1170-2.

Zaman, Z. and Leong, T.S. (1988): Occurrence of Paracamallanus malaccensis

Fernando and Furtado 1963 in Clarias batrachus and C.

macracephalus from Kedah and Parak, Malaysia. Asian Fish. Sci.,

2: 9-16.

Zarina Zaman (1990): Infection of a digenean Trematodes Orientocreadium

batrocoides in two species of genus Clarias collected from Kodah

State of Malaysia. Dhaka Uni. Stud. Part. E. Biol. Sci., 5 (1): 105-

114.

Zhang Yueyuan (1991): Three new species of trematode of Genus

Phyllodistomum (Trematoda: Gorgoderidae). Zoological research.,

vol. 12, No.4: 349-354.

Zietkiewicz, E., Rafalski, A., Labuda D. (1994): Genome fingerprinting by

simple sequence repeat (SSR) anchored polymerase chain reaction

amplification. Genomics 20: 176-183.

Zmeev, G.I. (1936): Les trematodes et les cestodes des poisons de I’ Amout

(Russian text). Parasitol. Sborn. Zool. Inst. Akad. Nank. SSSR

Leningrad (6): 405-36.

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