International Journal of Pharma and Bio Sciences IMPACT OF PLANT EXTRACT, COCCINIA GRANDIS ON DIGESTIVE TRACT MICROFLORA ABSTRACT

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1 Research Article Biotechnology International Journal of Pharma and Bio Sciences ISSN IMPACT OF PLANT EXTRACT, COCCINIA GRANDIS ON DIGESTIVE TRACT MICROFLORA HEMALATHA.J*, KALEENA.P.K AND K. VALIVITTAN *Department of Biotechnology, St Peter s University, Avadi, Chennai , Tamilnadu, India. Dept. of Advanced Zoology & Biotechnology, Presidency College, Chennai, Tamilnadu, India. ABSTRACT Secondary metabolites of plant extracts are effective to maintain the health status of the living organisms, mainly on the digestive system of animals. Coccinia grandis is a source of nutrient with abundance of effective microorganisms to digest the consumed foods on one hand and improve the health status of consumer on the other hand. With this background in the present study effectiveness of various organic solvent extracts of C.grandis on the intestinal microbial population was screened. Among the various extracts of Coccinia grandis studied ethanol extract of Coccinia grandis exhibited a profound increase in THBP in all the regions of the gut and the percentage increase was 30, 13 and 13 at 75 ug/ml and was 43, 40.5 and 45 at 100 ug/ml in foregut, midgut and hindgut respectively. In the case of ethanol extract Coccinia grandis at the concentration of 100 ug/ml Pseudomonas, Proteus, Achromobacter, Flavobacterium sp and E.coli were found to increase in colony wise. No much variation was observed in the microbial population of Staphylococcus where as a profound elimination of microbial population in case of Micrococcus, Streptococcus and Bacillus KEYWORDS: gut flora, microbial analysis and C.grandis. HEMALATHA.J *Dept of Biotecchnology,St. Peter s University, Avadi, Chennai Tamilnadu, India *Corresponding author B

2 INTRODUCTION C.grandis is an evergreen shrub, spread widely throughout the tropics because it thrives in a variety of soils, propagates easily, and bears fruit relatively quickly. The phytochemical compound of the C.grandis has enhanced the digestibility 2. Plants and plant extracts are effective mainly on the digestive system of animals. They function either by wiping out the pathogenic microflora in the digestive system or increasing the concentration of microbial population in the digestive system that contributes to improved digestion and absorption of nutrients 8. The stimulation of beneficial bacteria generates an effective protection against pathogenic microorganisms and a balanced intestinal microflora 7. The significant decrease in the numbers of pathogenic microorganisms is accompanied by the increase in the amount of mucous Ig A thereby protecting the intestinal mucosa against pathogenic microorganisms 5.Indeed many of the beneficial actions of plant extracts are thought to be via the additive and or synergistic actions of several secondary metabolites acting at a single or multiple target sites 1. Probably the most extensively researched and reviewed of the plant secondary metabolites in terms of both their anti nutritional and potential beneficial effects on ruminants are the tannins 3. Many plant secondary metabolites possess hormone like bioactive properties. Most data on their systemic effects come from medical and pharmaceutical research. Very little research, if any at all has investigated the possibilities of exploiting these properties to manipulate peripheral metabolism for the benefit of animal performance (Yoshiki et al., 1998 and Henry Greathead, 2003). With this background in the present study effectiveness of various organic solvent extracts of Coccinia grandis on the intestinal microbial population of fish was screened. MATERIALS AND METHODS The plant sample (C.grandis) was collected in early morning and shadow dried in preparation of extract. Ten gram of powdered plant material of Coccinia grandis were taken in clean sterile Soxhlet apparatus and extraction was done with 100 ml of different solvents (low polar to high polar) like as hexane, butanol, ethanol, chloroform and water. After extraction the extracts were dried at room temperature until extract reach into solid form. From the solid extract suitable concentrations were made using Dimethyl sulfo-oxide (DMSO) for further analysis. Laboratory breed fish Cyprinus carpio of either sex (2 months of age; weighing 25-30g) were used to evaluate the gut flora activity of different organic extracts of the Coccinia grandis. Fishes were housed under standard condition of temperature (27 C), 12h/12h light /dark cycles and fed with balanced pellet diet (Lipton, India Ltd). The acclimatized animals were divided into control (no treatment) and test groups (treated with plant extracts). After treatment of 21 days the test animal s digestive system (stomach, intestine and rectum samples) was collected for microbial analysis. Total heterotrophic bacterial population The total heterotrophic bacterial population was enumerated by pour plate technique using nutrient agar medium. The stomach, intestine and rectum samples were homogenized individually using a known volume of sterilized distilled water to make serial dilutions. After serial dilution with precaution, one ml of aliquots of appropriate dilutions of the sample was pipetted out into sterile petridishes and 15 to 20 ml of sterile nutrient agar medium were poured. The medium and the inoculums were thoroughly mixed using turn table and the medium was allowed to solidify. Duplicate plates were also maintained. The numbers of bacterial colonies were counted after 48 hrs of incubation. The bacterial populations were expressed as number of colony forming units (CFU) per gram samples analyzed. Generic composition of gut micro flora Representative of morphologically dissimilar well isolated colonies were selected at random from the nutrient agar plates of stomach, intestine and rectum samples. The selected colonies were sub cultured to check B

3 purity after noting morphology and pigmentation of colony. Then the pure bacterial strains were again sub cultured in nutrient agar slants. The slant cultures were stored at 4 o C in refrigerator and periodical subculturing was done to maintain the viability of the bacterial strains. The bacterial cultures were identified up to generic level by employing the scheme of Simidu and Also 4. All the experiments were carried out in triplicates. RESULTS AND DISCUSSION The THBP count in different regions of gut of control and plant extracts treated fish was given in Figure 1. When compared the THBP in the different regions of gut due to various extracts of Coccinia grandis with that of control, microbial population was found to be increased in fish administered with various extracts of Coccinia grandis. Among the various extracts of Coccinia grandis studied ethanol extract of Coccinia grandis exhibited a profound increase in THBP in all the regions of the gut and the percentage increase was 30, 13 and 13 at 75ug/ml and was 43, 40.5 and 45 at 100ug/ml in foregut, midgut and hindgut respectively, where as in fish administered with butanol extract of Coccinia grandis (100ug/ml) the THBP was found to decrease in mid and hind gut by 4 and 7 percentages. A moderate level of increase in THBP was observed in the various gut regions of the fish due to all other extracts of Coccinia grandis (Table 1). Total heterotrophic bacterial population count in the different regions of the gut included symbiotic as well as pathogenic microbes. Symbiotic microbes have probiotic role and promote digestive ability by producing microbial enzymes. The increase in THBP in different regions of the gut due to Coccinia grandis extract treatment indicated the increase in beneficial microbes and was substantiated by the generic composition of the gut microflora of the present study. The increment of beneficial gut microflora had enhanced the digestive ability, food consumption, energy and overall health of the animal. B

4 Figure 1 Total heterotrophic bacterial population in the alimentary tract of fish maintained as control and fish exposed to various extracts of fruits of Coccinia grandis. In the plant extract treated fish, the populations of the following bacterial genera were not much affected. They were Pseudomonas, Proteus, Bacillus, Flavobacterium and Micrococcus (Table 2). In the case of ethanol extract Coccinia grandis at the concentration of 100 ug/ml Pseudomonas, Proteus, Achromobacter, Flavobacterium sp and E.coli were found to increase in colony wise. No much variation was observed in the microbial population of Staphylococcus Sp. where as a profound elimination of microbial population in case of Micrococcus, Streptococcus and Bacillus In fish due to butanol extract of Coccinia grandis 33% elimination of Staphylococcus was observed. In fish administered with all extracts of Coccinia grandis showed slight increase in microbial population particularly Pseudomonas, Bacillus sp and Flavobacterium while a profound increase of Proteus sp and Achromobacter sp was noticed. Regarding the microbial population B

5 E.coli and Aeromonas sp no variation was observed in the various regions of the gut of fish after administered with plant extracts. It is obvious from the present study that the ethanol extract of Coccinia grandis had a good bacterial activity as recorded by Tullanithi et al 6., (2010) in A.aspera. Table 1 Total heterotrophic bacterial population in the alimentary tract of fish maintained as control and fish exposed to various extract of fruits of Coccinia grandis in fish. CONCLUSION Group of animal Extract concentration (µg/ml) Bacterial density (CFU/g) Fore-gut Mid-gut Hind-gut Control Hexane Butanol Ethanol Chloroform Water Table 2 Generic composition of bacterial strains isolated from fish maintained as control and fish exposed to various extract of Coccinia grandis Experimental Animal Plant material (ppm) Micro-coccus Strepto-coccus Staphylo-coccus Pseudo-monas Bacterial genera Bacillus Achrom-obacter Control Hexane Butanol Ethanol Chloroform Water Among the various extracts of Coccinia grandis studied ethanol extract of Coccinia grandis exhibited a profound increase in THBP in all the regions of the gut. From this the plant extract of C. grandis show some active principles to enhance the beneficial microbes in host system. These active principles would be helpful to prepare the immunostiumulant drugs. Flavo-bacterium Proteus Aero-monas Escher-ichia Total no. strains B

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