Effect Of Different Levels Of Probiotic Additive Biology Essay

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To evaluate the effect of different levels of probiotic additive on growth indices and body composition of juvenile O. niloticus and L. rohita, a 60 days experiment will be conducted. Feed will be given at two feeding levels 4 and 6% of body weight each level contain 0(control), 2, 4, 6 and 8% of probiotic Biogen (manufactured by SEARLE Pakistan limited). 300 juvenile O.niloticus and L. rohita (150 of each specie) will be distributed in 10 glass aquarium (3 replicates of each treatment).Water of aquaria will be changed after two days and feed remains and feces will be siphoned to maintain good water quality. Fish growth parameters viz; body weight and total length will be measured on weekly basis. Growth performance in terms of weight gain (WG), specific growth rate (SGR), feed conversion ratio (FCR), survival rate (SR) and body composition (moisture, carbohydrate, crude protein, lipid and ash content) will be analyzed. Water sample from aquarium will be collected and analyzed for physical and chemical characteristics.The data thus obtained will be subjected to statistical analysis using ANOVA and correlation.



(Synopsis for M.Phil Zoology)

TITLE: Effect of different levels of probiotic additive on growth indices and body composition of juvenile Oreochromisniloticus and Labeorohita.

Date of Admission : 21-11-2011

Date of Initiation : Ist March, 2013

Probable Duration : 60 Days


Name of student : MisbahNoureenDogar

Regd.# : 2009-ag-938

Supervisor : Dr. Abdul Mateen


Dr. Abdul Mateen (Chairman)

Dr. Muhammad Afzal (Member)

Dr. ZahedMahmood (Member)


Aquaculture imparts a major role in economy of many countries providing new employment opportunities to improve the economic situation for the people in these regions but its development faces many problems like disease, low production. These problems are due to aquatic animals exposed to stressful conditions, poor nutrient balance in the diet of fish and poor environmental conditions. Disease, mortality and poor growth is mainly due to physiological stress (El-Harounet al., 2006, Rollo et al., 2006).

Hormones, antibiotics, some salt compounds and plant extracts have been used for disease control and growth enhancement.But aquaculture faces troubles due to the adverse effects of antibiotics as they accumulated in a tissue that is why antibiotics have been prohibited in many countries due to their carcinogenic effects in telosts (Gatesoupe, 2007,Nayaket al., 2007).Antibiotics are also prohibited because pathogens gain resistance against antibiotics and these pathogens are responsible for spreading disease especially in stressful conditions(Suzeret al., 2008).

Use of probiotics in fish diet gave a new idea in aquaculture. Probiotics are live microorganisms, which gave better microbial balance in intestine, better utilization of feed and also enhance growth of organisms (Verschuereet al., 2000).Fish fed with diet having probiotics showed better growth rate because of better nutrient assimilation as well as reducing mortality and increasing resistance against pathogens(Irianto and Austin, 2002).

Several studies showed that probiotic in diet improve the health of larval and juvenile fish increase the disease resistance, body composition and growth performance. Fish like African cat fish, Clariasgariepince, Tilapia, O.niloticus (Lara-floreset al., 2003), Indian major carp,LabeorohitaHam (Kumar et al., 2006) andSeabass, Dicentrarchuslabraxshowed better growth, beneficial microorganisms in gut, increase digestive enzyme activity and feed utilization (Watson et al., 2008).

Oreochromisniloticusand Labeorohita are important freshwater fish species cultured in world. Considering the economic value of carp farming and also limiting studies of probiotics in juvenile carp diets, this experiment is planned to determine thedifferent levels of probiotic additives on growth indices and body composition of juvenile oreochromisniloticus and Labeorohita.

Review of Literature

Literature regarding the present work is reviewed as below:

Kumar et al. (2006) studied the probiotic incorporation in diet of Indian major carp, Labeorohita,by using Bacillus subtilis, a Gram-positive, aerobic, endosporeforming bacterium The diet containing B. subtilis showed maximum per cent survival (87.50%), weight gain (35.5%), TLCs (3.23Ã- 104 cells mm-3), haemoglobin content (7.4 g%), total protein (2.37 g dL-1) and globulin content (1.28 g dL-1) during the pre-challenge. Enzymes showed higher activities during post challenge. The result suggests that B. subtilis can be used as a commercial product for use in aquaculture.

Soltan and El-Laithy (2008) studied the effect of addition of different probiotics (Bacillus subtilisand Biogen®), spices (garlic or fennel) and also a combination of B. subtiliswith garlic or fennel in the diets of Nile tilapia. 420 Nile tilapia fry were distributed in seven treatments, 3 replicates of each treatment. Control (D1),others diets were supplied by additives, B. subtilis(D2), 0.2% Biogen® (D3), 1% garlic (D4), 1% fennel (D5), B. subtilis+1% garlic (D6) and B. subtilis+ 1%fennel (D7).The results of this study showed that diets with probiotics and spices significantly improved survival rate of Nile tilapia and diet containing combination of probiotics and spices showed best survival rate but not significantly differ from diet containing probiotics and spices alone. Feed utilization, Growth performance including body weight (BW), total length (TL), specific growth rates (SGR), feed conversion ratio (FCR) were higher in all treatments other than control.

Marzouket al (2008) studied the influence of types of probiotics on growth performance and intestinal microbial flora of Nile tilapia. Growth was determined in terms of body weight gain (WG), feed conversion ratio (FCR), and protein efficiency ratio (PER). Fish was distributed into three experimental groups. Control was fed with basal diet without any probiotics,second group was fed with dead Saccharomyces cerevisae.At the end of the experiment fish gains weight 5.8±0.25, 10.7±1.58 and 12.3±14.5. FCR 5.8, 3.0 and 4.7 for control, group 1 and group 2 respectively. Results indicated that fish fed with live Bacillus and Saccharomyces cerevisae gave best growth as compared to other groups.

Essaet al. (2010) evaluated the effect of different probiotic groups, Bacillus subtilis, Lactobacillus plantarum, and a mixture containing bacterial isolates (B. subtilisand L. plantarum) and a yeast, Saccharomyces cerevisiaeon growth performance, feed utilization and digestive enzyme activity of Nile tilapia. Five experimental diets, control diet was without probiotics Supplement diets were formulated to contain 107 CFU/g diet from Bacillus subtilis(D1),Lactobacillus plantarum (D2), a mixture containing bacterial isolates (B. subtilis&L. plantarum) (D3), while diet (D4) was formulated to contain 104 CFU/g diet of a yeast, Saccharomyces cerevisiae . Results showed that diets with probiotics improved fish growth,feed utilization as compared to control diets. S.cerevisiaeshowed higher amylase activity than the fish fed with control diet, other probiotics with exception of S.cerevisiae showed higher amylase, protease and lipase activity. These results recommended that Nile tilapia showed better growth performance and digestive enzyme activity if fed with diets containing probiotics.

Lara-Flores et al. (2010) studied the growth performance and intestinal enzyme activity in Nile tilapia as affected by two types of probiotics,a mix of two bacteria and yeast. Three diets one was a bacterial mixture containing Streptococcus faeciumand Lactobacillus acidophilus; a second was supplemented at 0.1% with the yeast Saccharomyces cerevisiae; and a third, was supplemented with a control diet without supplements. Two additional diets were formulated to contain 27% protein to serve as a stress factor. They were supplemented at 0.1% with either the bacterial probiotic mix or the yeast. The diets were fed for 9 weeks to tilapia fry housed in 20-L tanks at two densities: a high density of 20 fry per tank as a stress factor; and a low density of 10 fry per tank. Every week an organism was selected from each tank for the enzymatic analyses of unicellular protein, alkaline phosphatase, disaccharidases and peptidase. Results indicate that the fry fed with diets containing probiotic supplement exhibited greater growth rate than those fed with control diet. Of the four probiotic treatments, the 40% protein diet supplemented with yeast produced the best growth performance and feeding efficiency. This was attributed to an increase in the alkaline phosphatase activity, suggesting that yeast is an appropriate growth-stimulating additive in tilapia cultivation.

Noveirian and Nasrollahzadeh (2011) studied the growth response of common carp,Cyprinuscarpio as fed by different levels of biogen probiotics additives. Four practicle diets, one is control without biogen probiotic,other diets containing 0.1, 0.2, and 0.3% biogen were used to feed fish. Seventytwo juvenile common carp were distributed in 12 fiberglass tanks. During 60 days of experiment fish fed with probiotics showed higher body weight (BW), specific growth rate (SGR), protein efficiency ratio (PER), feed conversion ratio (FCR) and survival rate (SR) in comparison with control diet. The best results in food efficiency and growth performance were observed in fish fed diets containing 0.3% of Biogen probiotic that were significantly different with that in other treatments (P < 0.05 ). Growth performance did not differ significantly (P>0.05) in fish fed diets containing 0.1 and 0.2 percent of probiotic. Fish carcass composition in the diets receiving probiotic improved from initial values i.e. higher levels of protein and lipid with lower levels of moisture and ash content and showed significant difference (P< 0.05) only with the basal control diet. There were no significant differences (P > 0.05) in body composition between the treatments which received probiotic. Feeding juvenile carp with diets containing Biogen probiotic improves growth performances and feeding efficiency in them; the optimum addition of probiotic is 0.3 g/100gof diet.

Giriet al (2012) studied the effect of probiotic(Pseudomonas aeruginosa) VSG-2 on the innate immunity and disease resistance of Labeorohita. Fish was fed with four experimental diets, one was control, Second contains VSG-2 at 105, third 107 and fourth group with 109 CFU/g showed higher survival rates against Aeromonashydrophila infection. VSG-2 at 107 CFU/g can increase immunity and disease resistance in L.rohita as compared to other groups.

Materials and Methods

The 60 days experiment was conducted in Saline Fisheries Laboratory Department of Zoology and Fisheries, University of Agriculture Faisalabad to evaluate the effect of different levels of probiotic additives on growth indices and body composition of juvenile Nile tilapia, Oreochromisniloticus and Labeorohita.Three hundred OreochromisniloticusandLabeorohita will transfer to a laboratory and acclimated to the basal diet for 14 days.150 Nile tilapia and 150 L.rohita distributed to 30 aquaria, 10 fish 5 of each species in each aquarium.Water of aquaria will be changed after two days to maintain good water quality. Based on the feeding behavior of juvenile fish, feed will be given two times a day; feed will be adjusted at 4 and 8% of body weight. Fish will weighed once a week and daily ration will adjusted accordingly. The control diet is without feed additives (Biogen), while in other treatments diet will be supplemented by 2, 4, 6 and 8% of biogen.

Meat samples from each experimental aquarium will be collected and freezed for the proximate composition analysis of cultured fish species in terms of moisture, crude protein,total fats, and carbohydrates following Association of official Analytical chemist (A.O.A.C.1995).

The data will be analyzed through statistical analysis by following Steel et al. (1997) through Micro Computer. The comparisons of weekly based average values for various parameters were analyzed by using Analaysis of Variance (ANOVA) and comparison of means by Duncan's Multiple Range test with repeated sampling. Correlation analysis was also performed to find out the relationship among various physico-chemical characteristics.