“THE EFFECT OF PHYTOCHEMICALS OF ALLIUM SATIVUM ON …

Research Paper E-ISSN NO : 2455-295X | VOLUME : 5 | ISSUE : 7 | JULY 2019

I N T E R N A T I O N A L E D U C A T I O N A L S C I E N T I F I C R E S E A R C H J O U R N A L

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“THE EFFECT OF PHYTOCHEMICALS OF ALLIUM SATIVUM ON THE BACTERIAL INFECTION IN TILAPIA FISH (OREOCHROMIS

MOSSAMBICUS)”

SAJAD AHMED 1 | DEEPIKA BHARGAVA 2 | VIPIN VYAS 3

1 DEPARTMENT OF ZOOLOGY AND APPLIED AQUACULTURE BARKATULLAH UNIVERSITY, BHOPAL. 2 DEPARTMENT OF ZOOLOGY AND APPLIED AQUACULTURE BARKATULLAH UNIVERSITY, BHOPAL. 3 DEPARTMENT OF ZOOLOGY AND APPLIED AQUACULTURE BARKATULLAH UNIVERSITY, BHOPAL.

ABSTRACT:

Aquaculture is emerging as the fastest growing food producing industry in the world and Tilapia fish plays a vital role in fulfilling the demand for fish food consumption. However the intensive culture of Tilapia fish led to the outbreaks of various bacterial diseases. Which directly affect the economic loss to the production of aquaculture. And it is estimated that industries at worldwide suffered the loss in billions of dollars annually. The antibiotics are used widely in the treatment of diseases in fishes. But due to excessive use of antibiotics the fishes led to the development of antibiotic- resistant pathogens. This doesn’t allow the fishes to be cured easily. So to overcome this problem we used Allium sativum as a phytochemical to treat the fishes so that the disease can be cured without any harmful effect. The extract is prepared from the garlic powder and then adds it to the feed. It automatically cure the fishes and protect them also from all kind of bacterial diseases.

KEYWORDS:

AQUACULTURE, ANTIBIOTICS, BACTERIAL DISEASE, PHYTOCHEMICAL, GARLIC EXTRACT.

INTRODUCTION

Global population increases and food demand have been motivating factors leading to additional expansion intensive animal and fish production. Recently, many attempts advocated the expansion in fish farming to optimize the real need for animal protein. Aquaculture has become a key component of the animal health industry due to continued expansion of cultured fish and shellfish species. Aquaculture is the fastest growing industry around the world with about 80 million tonnes being produced annually. Aquaculture is therefore an emerging industrial sector which requires continued research with scientific technical development and innovations. Bacterial diseases are responsible for heavy mortalities in both culture and wild fishes throughout the world and most of the causative microorganisms are naturally occurring opportunist pathogens. Attempts to control or prevent such devastating outbreaks using conventional antimicrobials and other chemotherapeutants have been generally unsuccessful.

The uncontrolled and repeated use of antibiotics to treat bacterial infection has in some cases led to the development of antibiotic resistant pathogens. Considering the potential threat of diseases on human and animal health issues associated with the use of antibiotics Management should therefore concentrate on environmental friendly preventative methods such as the use of plants as antibacterial agents and immuno-stimulants.

MATERIAL AND METHODS

GARLIC (ALLIUM SATIVUM)

Garlic is a bulbous perennial plant growing up to 1 meter in height. It grown from an underground bulb which sends shoots up in the air. Garlic bulb is divided into papery white clones (their number range from six to twenty).The leaves are long and flat, opposite and erect, with a crease down the middle. Flowers are white and starry. The garlic plant has 6-12 long, narrow and flat like grass leaves. The most important part of this plant for medicinal purpose is the bulb. The bulb consists of numerous bullbats, known as clones held between the papery membranes and enclosed with in a lintiest skin as a sac. Each bulb is made up to 20 cloves, and each clove weighs about 1 gram, Allium sativum has an annual life cycle. The main phytochemical in garlic is allicin. When garlic is damaged or crushed the sulphur components are transformed in different organ sulfur compounds. Enzymes in garlic will convert alliin into allicin, which has antimicrobial action. Garlic and garlic preparations have many claimed effected. Reduction of risk of cancer and heart diseases, stimulation of the immune system. Allicin is garlic’s defense mechanism against attacks by pests. The enzyme allinase converts the chemical alliin to allicin, which is toxic to insects and microorganisms. The antimicrobial activity of allicin was discovered in 1044 by Cakallite. Allicin extracted from garlic looses beneficial properties within hours and turns into other sulphur containing compounds. Diallyl trisulfade, which is similar to allicin but is chemically produced is stable and is used for treatment bacterial, fungal and parasitic infections.

METHOD OF EXTRACTION FROM ALLIUM SATIVUM

STEPS:



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Purchased fresh garlic from market.

Peeled it and then rinsed with water.

Cut into small pieces with sterile knife.

Air dried under a shade for 14 days at room temperature.

Garlic bulbs were reduced to coarse powder.

10gm of garlic powder was soaked in 100ml of ethanol 80% and methanol 96% for 30 hours.

Filtered with what man no. 1 filter paper.

Used a rotary evaporator and finally dried in oven at 40°c for 24 hours.

100mg of each dried extract was dissolved in an appropriate vol. of DMSO and the vol. reached to 1ml to get the final conc. Of 100gm/ml.

Then store the extract in a dark bottle at 40°c for further use.

STREPTOCOCCUS IN TILAPIA

Streptococcus is a major problem for tilapia in aquaculture and can rapidly spread throughout the farm. It can lead to mass-death and unlike many other diseases a streptococcus infection can be fatal even for large and otherwise healthy fishes. Weakened fish is especially susceptible to the illness.

A tilapia can catch streptococcosis from an infected fish as well as from the environment. In aquaculture streptococcosis is usually caused by the bacterium Streptococcus agalactiae. Streptococcosis is usually caused by the bacterium Streptococcus iniae also occurs in fish farms but such outbreaks are usually not as lethal as those caused by Streptococcus agalactiae.

SYMPTOMS OF STREPTOCOCCOSIS IN TILAPIA

1. Eye problems.

2. Abscesses.

3. Skin hemorrhages.

4. Ascites.

5. CNS Problems.

FORMULATED FEED

1. In culture practices where food is not sufficient, artificial feed otherwise called supplementary feed is necessary. However for optimum growth and metabolism of animal a well balanced nutritious diet is required.

2. The nutritionally balanced diet contains protein fat carbohydrates, minerals, vitamins and specially carotenoids because caroteniods are the primary source of the pigmentation.

3. Properly formulated prepared feeds have a well balanced energy to protein ratio.

4. Feed is available in a variety of size ranging from the fine crumbles for small fish to large (1/2 inch or larger) pellets. The pellets size should be approximately 20% to 30% of the size of mouth of fish.

FEED INGREDIENTS

• Wheat flour

• Rice bran

• Fish meal

• Soya bean oil or oil cake

Because these ingredients are easily available and are cheapest also as compare to the other. And the amount of these ingredients depends upon their proteinous value. The feed should be comprises approx. 30% protein.

TABLE NO.1 PERCENTAGE COMPOSITION OF BASAL DIET.

Ingredients Diet

Fish Meal 43.4%

Wheat Flour 28.3%

Rice Bran 28.3%

Total 100%

PREPARATION OF CONTROL DIET:

1. Artificial diet was prepared to feed the fishes with various feed ingredients. Fish meal, wheat flour were finally ground and proximate analysis was performed using Pearson square method.

2. Based on this analysis, one practical diet was formulated containing 30% protein. Vitamin and mineral mixes were then added by continuous mixing.

3. Distilled water (27°C) was slowly added to the

feed to get desired consistency for pelleting. This was then extruded in hand pelletizer using a 1mm die and dried for 4-6 hours below 45°C and subsequently air-dried overnight to reduce moisture content less than 10%.

Dried feed was chopped into pellets in a blender passed through sieves to ensure a homogenous particle size(1.0-2.0mm). The feed was further stored at room temperature in air tight jars.



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PREPARATION OF EXPERIMENTAL DIET:

1. The experimental diets impregnated with Garlic extract were prepared by replacing fishmeal in the basal diet by Garlic extract at the rate of 5%.

2. Different percentages of fish meal were used for feed formulation while wheat flour and vitamin-mineral mixture contents were the same as in the four formulated feeds.

3. The feed was pelletized with hand pelletizer, dried in hot air oven at 50°C and stocked in air tight jars.

TABLE-2: PERCENTAGE COMPOSITION OF VARIOUS INGREDIENTS IN EXPERIMENTAL DIETS/100G.

PREPARATION OF EXPERIMENTAL DIET /100g DIETS FISH MEAL WHEAT FLOUR RICE BRAN GARLIC EXTRACT

FEED FOR CONTROL

GROUP 43.40% 28.3% 28.3% 0%

EXPERIMENTAL FEED

WITH (5%GARLIC

EXTRACT)

38.40% 28.3% 28.3% 5%

OBSERVATIONS

WEIGHT AND SURVIVAL RATE ANALYSIS

At the termination of the experiment, highest value for average weight gain in the fishes was observed in feed with 5% Allium sativum which came out to be the

tremendous increase in the body weight as for as they overcome with these infection on the other side the fishes in the control group was observed with the great loss in the weight due to the bacterial infection.

TABLE.3 SHOWING OBSERVED WEIGHT (GM) OF OREOCHROMIS MOSSAMBICUS USING DIFFERENT FEEDS

Aquarium Feeds

Observed average weight in gm Initial Average Weight

7th day

14th

day

21st

day

28th

day

35th day

42nd

day

49th

day

60th day

1st

Control Feed for infected fishes

33.6 gm 32.0gm 30.2gm 28.6gm 27.2gm 26.0gm 24.6gm 23.2gm 22.2gm

2nd

Feed with 5% Allium sativam

30.2gm 31.8gm 33.2gm 34.8gm 36.4gm 39.0gm 41.2gm 44.0gm 46.2gm

TABLE .4 SHOWING SURVIVAL RATE OF OREOCHROMIS MOSSAMBICUS AT DURING EXPERIMENTAL PERIOD.

Aquarium Feeds Survival rate (%)

1st Control feed 75%

2nd Feed with 5% Allium sativum 100%

FIG.1. SHOWING VARIATIONS IN WEIGHT IN THE FISHES IN CONTROL AND EXPERIMENTAL GROUP.



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FIG.2 BAR GRAPH SHOWING SURVIVAL RATE OF FISH IN CONTROL AND EXPERIMENTAL GROUP.

RESULTS

Dietary supplementation with garlic extract for 60 days prior to challenge significantly reduced the Streptococcus sp. Infection.

The result of the study reinforce the growing view that dietary supplementation of garlic extract is beneficial to fish health by conferring protection against pathogens,

As earlier we took two aquariums for this experiment, and we took 16 fish species of Oreochromis mossambicus and put them in the aquariums (8 species each) and named them

1 Control Group 2 Experimental Group

CONTROL GROUP EXPERIMENTAL GROUP

FIG.3. SHOWING THE AQUARIUM OF CONTROL GROUP. FIG. 4 SHOWING THE AQUARIUM OF EXPERIMENTAL GROUP.

And treat the fishes of both the groups with normal feed for two weeks This allows them to adapt the environment completely. On the other side we brought water from the same lake and culture bacteria with that contaminated water

Once the fishes adapt the environment completely add the prepared bacterial media in both the groups and kept them for at least 7 days (one week) and allowed the fishes to come in contact with bacterial infection. After seven days we observed that all the fishes was infected as shown in the below pics.



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FIG. 5. SHOWING THE BACTERIAL INFECTION (CONTROL GROUP) FIG. 6. SHOWING BACTERIAL INFECTION ( EXPERIMENTAL GROUP)

FIG 7. SHOWING THE INFECTED FISHES OF CONTROL GROUP TREATED WITH NORMAL FEED.

FIG 8. SHOWING THE FISHES OF EXPERIMENTAL GROUP TREATED WITH THE FORMULATED FEED (5%ALLIUM SATIVUM)



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CONCLUSION

• Fish is one of the cheapest and most promising sources of animal protein. People can easily digest 93.2% of fish protein and this shows the importance of fishes for us. By knowing the demands of the public some people start culturing it to increase the fish production.

• But there are some dreadful diseases and infections also which effect the production of fishes badly. The most common disease is known as bacterial infection. In Tilapia fish (Oreochromis mossambicus) the bacteria species Streptococcus agalactiae is commonly found.

• In this dissertation our main motto was to overcome with this problem.

• So we brought 16 fishes of species Oreochromis mossambicus and put them in two different aquarium namely control group and experimental group 8 species each and allowed them with the bacterial infection which was mostly seen in them.

• Then treat both the group with different feeds, The control group was fed with the normal feed but experimental group was fed with the 5% Allium sativum feed.

• And at the end of our experiment we observe that the phytochemicals of Allium sativum was highly effective on the bacterial infection and the result showed that variation in the weight of the fishes also.

• The fishes treated with 5% Allium sativum feed gain their weight normally and also overcome with all type of infection and it strengthen their immune system. It also increase the survival of the fishes. Which directly affect the production of the fishes.

• After this successful experiment we recommend this formulated feed (5% Allium sativum) to all the fisherman because it is very cheap and doesn’t had any side effect .

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“THE EFFECT OF PHYTOCHEMICALS OF ALLIUM SATIVUM ON …