direct fed microbial

Aim: To study biological response of the isolated strain (Direct fed microbial- DFM) of Enterococcus spp. in broilers at different dose rates. 
 
Materials and Methods: Various treatments consisted of T0 : Control (Culture medium); T1 - isolated direct fed microbial namely Enterococcus faecium (2.8 x 108 cfu/kg feed); T2 - Enterococcus faecium (4.8 x 108 cfu/kg feed), T3 - Enterococcus faecium (6.8 x 108 cfu/kg feed) and T4 - Enterococcus faecium (8.8 x 108 cfu/kg feed). Growth attributes and feed consumption was recorded at regular intervals. Randomly 6 birds per treatment were chosen for blood collection and for estimation of calcium, phosphorus and cholesterol concentration in blood. Representative sample of birds were slaughtered and caecal contents were collected under sterile conditions to study microbial flora. 
 
Result: Dose specific results for growth attributes, FCR, plasma concentration of calcium, phosphorus and cholesterol, and microbiological evaluation of caeca were obtained after supplementation of isolated DFM. Enterococcus spp. supplementation at low doses (2.8 x 108 cfu) resulted in significant increase in weight gain and better FCR. Low dosage supplementation of Enterococcus spp. also increased calcium concentration and lowered cholesterol in blood. Birds in treatment T4 supplemented with high dose of Enterococcus spp. performed poorly as compared to control. All treatments had high ratio of gram positive to gram negative bacteria. 
 
Conclusion: It was concluded that isolated strain of Enterococcus acted as probiotics only at low dosage (2.8 x 108 cfu). Further it was found that overall biological performance was negatively correlated to higher dose (8.8 x 108 cfu) of supplementations of Enterococcus spp. Micro floral study of caecal content indicated that the strain was able to establish itself in the gut.


Introduction
reached inside the gastrointestinal tract.The intestinal bacterium Enterococcus faecium has been used for The term Probiotics is defined by United Nation more than a decade as a probiotic strain.and WHO [1] expert panel as live organism which Hence, the aim of this study was to evaluate the when administered in adequate amount confers a use of Enterococcus faecium isolated from leopard health benefit on host.In recent years, probiotics have faeces supplemented at different dose rates as a emerged as major tool in improving the feed efficiency, probiotics in poultry birds.growth promotion and disease control in animals.Probiotics could be useful in decreasing the cost of

production, disease prevention and for better growth in
Experimental animals and ethical approval: Three food animals.Numerous studies till date have hundred (300) commercial broiler chicks of Vancob demonstrated increased weight gain and better feed strain were procured for the present investigation.conversion ratio as a result of probiotic supplemen-Experiment was ethically approved in accordance with tation in food animals [2][3][4][5][6].Probiotics exert positive laws and regulations.effect on host by improving the gut health [7,8].Further it also affects mineral absorption [9] and cholesterol Study design: The broiler chicks were divided into 5 concentration in blood [2,10].The action of probiotics groups with three replicates consisting of 20 chicks varies with the dose rate [11], therefore it is very each in a randomized block design.The feed was important to formulate dose for a particular probiotics formulated and prepared as per the BIS standards [12].microorganism so that suitable microbial balance is Perusal of the proximate composition ( Gram's stain and subsequently count was obtained for cfu/kg feed), T -Enterococcus faecium (6.8 x 10 there was no significant (P>0.05)difference between ml of sample was obtained employing plate count method.
the treatments.The FCR value was better for T and 1 highest for control but difference was again non-Method of administration of probiotics: Isolated microbial significant (P>0.05)among different treatments.culture was mixed with feed quota of a particular The difference in live weight gain and FCR in treatment before feeding every day and it was different treatments during the starter phase was also continued during entire period of the feeding trial.
found to be non-significant (P >0.05).Standard management practices were followed for No distinct effect of probiotic supplementation rearing.Different parameters such as weight gain and was found during pre-starter and starter phase (Table-2).feed consumed was recorded at regular interval.
During the finisher phase, highest live weight gain and Blood collection: Approximately 5 ml of blood was better FCR was observed in treatment T that was 1 collected and plasma separated from randomly chosen significant as compared to control and other treatments.6 birds per treatment for estimation of calcium, The live weight gain in treatment T was better (P 2 phosphorus and cholesterol.Calcium was estimated by <0.01) than T but comparable to T and T .The FCR Elmer 3100 (USA, 1982)] while inorganic phosphorus significant (P<0.01) as compared to all other and cholesterol were estimated by photometer 5010 V treatments including control.Only the birds in T 1 2.0 using analytical kits of Bayer diagnostics.performed better (P <0.01) than control, while in other groups it was statistically comparable during the Microbial count in large intestine: Representative finisher phase.sample of birds were slaughtered and caecal contents Overall evaluation of results also revealed that were collected under sterile conditions to study birds in Treatment T performed significantly better as microbial flora.Total bacterial count of the represen-1 tative sample from caeca of birds was done on nutrient compared to control and other treatments in terms of agar by plate count method.Serial dilutions were made gain in weight and FCR whereas birds in treatment T , 4 which was fed with high doses of probiotics, performed with direct fed microbial, whereas the count of Gram poor in terms of growth performance.
-ve bacilli in large intestine was highest in control as Thus, the perusal of results revealed that probiotics compared to all other treatments (Table-4).The above supplementation at different doses did not affect the pattern probably indicates that the DFM was able to performance of birds during pre-starter and starter establish itself in the GIT of broiler birds as compared phase but had significant effect during the finisher to control and excluded the dominance of Gram -ve phase.Probiotics beneficially affects the performance bacteria in the large intestine of broiler birds.This may of poultry [1, 5,15] but suitable microbial balance be be probably due to the reason that Enterococcus strain reached which positively affects the bird's performance possesses good colonization ability [23] also at a particular dose of probiotics.The overall trend supporting the fact that probiotics are able to lower the revealed that though probiotics supplementation at low colonization of coliforms and other gram -ve bacteria doses was beneficial in terms of weight gain and FCR.[24,25].higher doses may have detrimental effect on growth Conclusion performance of broiler birds.Similar results were obtained by other workers who found a negative Thus it may be concluded that feeding of isolated correlation of higher dose of probiotics with direct fed microbial (DFM) Enterococcus faecium performance of broiler birds, supporting the above from leopard faeces (Pathera leo) at a dose rate of 2.

3 8 different
bacteria types.cfu/kg feed) and T -Enterococcus faecium (8.8 x 10 4 cfu/kg feed).Statistical analysis: The data was analyzed by using one-way analysis of variance technique (ANOVA) as Isolation and characterization of probiotic: Direct fed per Snedecor and Cochran [14] by using SPSS version microbial Enterococcus faecium was isolated from 12. leopard (Patheraleo) faeces and characterized using Results and Discussion morphological and biochemical characteristics as proposed by Cowan et al. [13].Isolated probiotic strain Gain in weight and feed conversion ration (FCR): The Enterococcus faecium was grown in Elliker broth and gain in live weight of different treatments during the O subsequently stored at 4 C.The viability and pre-starter phase was almost similar to each other and concentration of the Enterococcus faecium culture per (AAS) [Perkin value was highest in treatment T and was highly 4

Table - 1
. Proximate composition of pre-starter, starter and finisher diet of broiler chicken.

Table - 3
. Blood plasma concentration of Ca, P and cholesterol in broiler chicks supplemented with Enterococcus strain PSE-Pooled standard error; Figures bearing different super scripts within a row differ significantly, ** P<0.01 Table-4.Microbial count in large intestine of broilers chicks supplemented with Enterococcus strain