“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.
Department of Zoology
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.
Department of Zoology
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
1) Senga rupchandensis n.sp.
2) Circumoncobothrium jadhavae n.sp
3) Genarchopsis paithanensis n.sp.
4) Allocreadium khami n.sp.
5) Orientocreadium striatusae n.sp.
CHAPTER-III HISTOPATHOLOGY
1) Senga rupchandensis n.sp.
2) Circumoncobothrium jadhavae n.sp.
3) Genarchopsis paithanensis n.sp.
4) Allocreadium khami n.sp.
5) Orientocreadium striatusae n.sp.
CHAPTER-IV
DNA FINGERPRINTING
1) Senga rupchandensis n.sp.
2) Circumoncobothrium jadhavae n.sp.
3) Genarchopsis paithanensis n.sp.
4) Allocreadium khami n.sp.
5) Orientocreadium striatusae n.sp.
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
phosphatase enzyme activity.
PLATE: 12) Alkaline phosphatase enzyme activity in Genarchopsis
paithanensis n.sp.
A) L.S. of whole parasite showing distribution of alkaline
phosphatase enzyme activity.
B) L.S. of anterior region showing distribution of alkaline
phosphatase enzyme activity.
C) L.S. of middle portion showing distribution of alkaline
phosphatase enzyme activity. .
D) L.S. of posterior region showing distribution of alkaline
phosphatase enzyme activity.
PLATE: 13) Alkaline phosphatase enzyme activity in Allocreadium
khami n.sp.
A) L.S. of anterior region showing distribution of alkaline
phosphatase enzyme activity.
B) L.S. of middle region showing distribution of alkaline
phosphatase enzyme activity.
C) L.S. of posterior region showing distribution of alkaline
phosphatase enzyme activity.
PLATE: 14) Alkaline phosphatase enzyme activity in Orientocreadium
striatusae n.sp.
A) L.S. of whole parasite showing distribution of alkaline
phosphatase enzyme activity.
B) L.S. of anterior region showing distribution of alkaline
phosphatase enzyme activity.
. C) L.S. of posterior region showing distribution of alkaline
phosphatase enzyme activity.
PLATE: 15) Acid phosphatase enzyme activity in Senga rupchandensis
n.sp.
A) L.S. of mature proglottids showing distribution of acid
phosphatase enzyme activity.
B) L.S. of mature proglottids showing distribution of acid
phosphatase enzyme activity (Magnified).
C) L.S. of gravid proglottids showing distribution of acid
phosphatase enzyme activity.
PLATE: 16) Acid phosphatase enzyme activity in Circumoncobothrium
jadhavae n.sp.
A) L.S. of mature proglottids showing distribution of acid
phosphatase enzyme activity.
B) L.S. of gravid proglottids showing distribution of acid
phosphatase enzyme activity.
PLATE: 17) Acid phosphatase enzyme activity in Genarchopsis
paithanensis n.sp.
A) L.S. of whole parasite showing distribution of acid
phosphatase enzyme activity.
B) L.S. of anterior region showing distribution of acid
phosphatase enzyme activity. .
C) L.S. of posterior region showing distribution of acid
phosphatase enzyme activity.
PLATE: 18) Acid phosphatase enzyme activity in Allocreadium khami
n.sp.
A) L.S. of anterior region showing distribution of acid
phosphatase enzyme activity. .
B) L.S. of middle portion showing distribution of acid
phosphatase enzyme activity.
C) L.S. of posterior region showing distribution of acid
phosphatase enzyme activity.
PLATE: 19) Acid phosphatase enzyme activity in Orientocreadium
striatusae n.sp.
A) L.S. of whole parasite showing distribution of acid
phosphatase enzyme activity.
B) L.S. of anterior region showing distribution of acid
phosphatase enzyme activity. .
C) L.S. of posterior region showing distribution of acid
phosphatase enzyme activity.
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
armatus (Lecepede, 1800).
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
armatus (Lecepede, 1800).
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:
1) Senga rupchandensis n.sp.
2) Circumoncobothrium jadhavae n.sp.
3) Genarchopsis paithanensis n.sp.
4) Allocreadium khami n.sp.
5) Orientocreadium striatusae n.sp.
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
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, mounting in DPX. Drawings were made with the help of
Camera Lucida and all the measurement are taken in millimeter.
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
Circumoncobothrium jadhavae n.sp.
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.
The present communication deals with the description of new species
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
mm. in length and 0.1283 mm. in breadth.
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
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, mounting in DPX. Drawings were made with the help of
Camera Lucida and all the measurement are taken in millimeter.
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
Host Channa striatus (Bloch, 1793)
Habitat Stomach
Locality Latur District, M.S., India
Date of collection May 2007.
Trematoda Rudolphi, 1808
Digenea van Beneden, 1858
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.
The present communication deals with the description of new species
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.
Trematoda Rudolphi, 1808
Digenea van Beneden, 1858
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.
The present communication deals with the description of new species
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
mm. in length and 0.2613 mm. in breadth.
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
measures 0.02245 mm. in length and 0.01604 mm. in breadth.
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.
Host Channa striatus (Bloch, 1793)
Habitat Intestine
Locality Kham River, Aurangabad District,
M.S., India
Date of collection Dec. 2008.
Trematoda Rudolphi, 1808
Digenea van Beneden, 1858
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.
Trematoda Rudolphi, 1808
Digenea van Beneden, 1858
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.
The present communication deals with the description of new species
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,
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 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
measures 1.1363 mm. in length and 0.6818 mm. in breadth.
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.
Host Channa striatus (Bloch, 1793)
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
armatus (Lecepede, 1800).
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:
Senga rupchandensis n.sp.
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.
Circumoncobothrium jadhavae n.sp.
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.
Genarchopsis paithanensis n.sp.
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.
Allocreadium khami n.sp.
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).
Orientocreadium striatusae n.sp.
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
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,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:
Senga rupchandensis n.sp.
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.
Circumoncobothrium jadhavae n.sp.
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.
Allocreadium khami n.sp.
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).
Orientocreadium striatusae n.sp.
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).
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 Channa striatus (Bloch, 1793) and Mastacembelus
armatus (Lecepede, 1800).
MATERIAL AND METHODS:
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.
RESULT AND DISCUSSION:
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.
Senga rupchandensis n.sp.
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.
Circumoncobothrium jadhavae n.sp.
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.
Genarchopsis paithanensis n.sp.
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.
Allocreadium khami n.sp.
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.
Orientocreadium striatusae n.sp.
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).
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 Channa striatus (Bloch, 1793) and Mastacembelus
armatus (Lecepede, 1800).
MATERIAL AND METHODS:
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.
RESULT AND DISCUSSION:
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
Order Pseudophyllidea
Carus, 1863
Family Ptychobethriidae
Luhe, 1902
Genus Senga Mastacembelus
Dollfus,1934 armatus
Species Senga rambaei n.sp. (Lecepede,1800)
Class Eucestoda
Wardle, McLeod &
Radinoky, 1974
Order Pseudophyllidea
Carus, 1863
Family Ptychobethriidae
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)
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