tf Reseach Jour of Biological Seenee Inflorescence Flore, August 2008p. 33-37 Prevalence of Migeobial Iniabi Bhakyaraj RJ. Vetriseh , Arun Sasi, N. Yogananth, A. Chanthura and S, ts from Cowshed and its € PG. and Research Department of Microbiology. 1J. College of Atts and Seer Padukkota “Deparment of Microbiology. Se 1622 404, Tail Nad In amalathayar College of Arts and Sehence (Wom) Department of Botany, Govt, Collegeof Arts and Science, Ka, Tamil Ni Marathi ann Nad, tia Corresponding author: rraibakye Received 8 Nah 2008 revered 3 Jane 2028: acpi uy 208 Abstract Cow dung con:sins vast quantities of pathogenic hactria and fungi Totaly four Nair ‘Sirepiococcus sp, Bacills sp, Closiradinn sp. Stapvtocor st emtectum, Aspergillus wus, Aspergillus versicolor, Asporglas ws, from cow dung with so and two dominant types of bacteria ie. Seprocaceus sp se, Fusarium semitectum, Aspergillus flavus were chosen for treatment with Na,Co, the exper tion spectra in both bacterial and fungal colonies compared with those in he 0 con showed ahighe soups. Tapers avons wore ists Pacts yp ad 1 fans ‘Keywords: Pathogenic, Cow dung, inhibition spectrum, No,CO, Optical density Introduction Our way of life has intensified the quantum of pollution. No place ean be called safe from pollution ‘What varies is the type of pollutant, waste-water pollution, ar pollution, radio active pollution and soon. Microbial pollution is the most important type of pollution for people inthe medical or paramedical field Microorganisms are ubiquitous in nature. ‘They are tnainly of two types, non-pathogenic and pathogenic “There are certain opportunistic pathogens, which given a chance ean produce diseases in human beings and animals. The mere presence of these microorganisms in ‘definite strength ean produce contaminantion in various ‘media (Mond ‘and undigested remains of plant food plus vast quantities 1981), Dungeonsists ofthe macerated of bacteria (mostly dead) as well a animal waste and bile products. such as broken down red blood ee! pigments. Ruminants produce fine textured dung of fibrous plant material whereas horses, witha less efficient system, produce much coarser excreta Dung decomposes rapidly because the macerated material has high nitrogen content, available aeration nd high water content that rected from fluctuations (Ing, 1989). Dany Fh nins, growth factors and mineral ions, some of which are metabolic by- products of microbes in herbivores" gut. Dung also contains a lara amount of readily avaiable carbohydrates (Hane 1988). All classes of fungi are found on dung, Zysomycetes usually appearing First followed by the 3Prevalence of Microbia Inabitants fom Cowshed and its Controt ‘Ascomycetes and finally the Basideomycets. Their distribution is influenced by’ the number of herbivores present in an area (Keendrik, 1985). Microbial inhabitants ofthe cowshed, cause diseases in human beings In event researches in California, these pathogens are controlled by using the Carbonate method. The use of Sodium Carbonate for inhibiting the growth and activity of pathogens and inthe management of disease has often tbeen emphasized (Anonymous, 2003). With this basic informa wn, the present investigation has been justifiably planned to isolate the ‘microbial inhabitants of cow sheds (including soil and ‘cow dung) and to study the pathogenic bacteria and fungi which cause diseases and the effect of chemicals such as Sodium Carbonate on the test organisms Materials and Methods In this study, for isolation.of microbial inhabitants, ‘cow dung and soil samples were collected from the ccowshed. The samples were placed in polythene bags. closed tightly and stored in efrigerator. The bacteria and fungal strains were isolated using diution-plating, technique and the diluted samples were taken up to 10°, 10% and 10° dilutions plated on Nutrient agar (NA) for bacteria and the dilutions 102, 10° and 10* plated on Sabouraud Dextrose Agar medium (SDA) for fing. The bacteria and fungal isolates were identified by wsing standard manuals, From the isolated colonies four predominant bacterial and fungal eotonies ephococens ™ sp, Bacillus sp, Fusarium semirectum and Aspersilas flavus ors selects fo For the evaluation of minimal inhibitory etfeet (MIE) of Na,CO, om the growtl of the test organism “Sireprococeus sp an Baciflus sp. the microbial bioassay method was used, Nutrient broth supplemented with magnesium was prepared and distributed in test tubes ‘each with 10m! of medium, On autoclaved. cooled one phof 24hrs culture of Sirprocacens sp and ‘Bacillus sp were added separately. ‘The different concentrations of Na,CO, were added separately 10 the ‘cooled nutrient broth to give the concentration of 1%. 246. 3% and 4%, All tubes were thes ineubated at 37°C for 2ahes. The Optical density (OD) value of each set ssured at 345nm and compared with the OD values of control. Nutrient broth without Na.CO, was used as control. For the fungi, different concer rations of Na,CO, (ae, 2%, 3% and 4%) were aukled separa cooled SDA ™ un. Fuser semitectun “Aspergillus flavus were inoculate! at the center of the SDA medium separately. ‘The plates were ine ec #2! subated at for 48h. The radial growth of the test organisms was noted, Absence of Na,CO, in SDA medium served as the contra Results and Discussion Rumen microbes are eapuble of transforming most lof the toxie eompninals acewmrates ay pants 18Rescach Joumal of Biological Science Tnftorescence Foret 1, August 2008, pp. 33-37 toxic beneficial compounds. Contrary to this, other types of microbes which were unable 10 metabolize the toxic compounds but convert the initially non-toxie metabolites ‘tw harmful products could also exist especially in summer (Dev * Were isolated and identified as Sireptococcus sp, Bacillus 1987). Number of bacteria and fungal colonies sp, Clostridium sp, Staphylococcus sp and Fusarium sdmitectum, Aspergillus flavus, Aspergillus versicolor Aspergillus ustus, Aspergillus sydowi from animal husbandry premises especially the cowshed, ‘The (wo dominant bacterial strains ie., Streptococcus sp, Bacillus sp and two fungi Fusarium semitectum, Aspergillus “flavus were selected for microbial bioassay method. As inthe present study, compost-inhabiting pathogens were isolated from cow dung and composted soil which includes Aspergillus niger, Trichoderma harzianam, Bacillus cereus and Bacillus subtilis (Muhannned, 2003). Microbial inhabitants of cowshed can utilize magnesium as their energy source. (Anonymous, 2003). Hence the medium was supplemented with Magnesiui “The predominant pathogens were treated with Na,CO, Which minimizes their growth. Many pathogens are present in the excretory matter and veterinary wastes especially cow dung. These pathogens utilize magnesium, which is present in cow dung. The pathogenic growth promoted by Magnesium was observed and these pathogens were treated with the carbonate method (Anonymous, 2003), The minimal inhibitory effect was obtained in Sireprococeus sp. treated with IYaaFthe Na,CO, whieh had the tity to inhibit microbial growth. The vale of OD at 1% was 0.054 whereas in control tubes 0.156 absorbance was noted. In Bacillus sp, 2% of Na,CO, (0.107) inhibit more microbial yrowih when compared with all other concentrations and with control tubes, (0.522). In this earbon te method: Hacillus sp was OE inhibi hly susceptible to Na,CO, noted in Table 1 and Pig 1 In fungus, the mean radial growth rate of Frsarian in the Na,CO, content in the SDA medium. The highest decrease was in 4% of Na,CO, than in other concentrations. The growth rate of Aspergillus flavus wis highly arrested even in 1% Na,CO, treatment. The resalts are shown in Table 2 and Fig 2. This preliniy screening shows that better sanitation with Na.CO, 1s needed in order to minimize the emergence and spread of pathogens in the eowshed. “Table 1. Effect of Sodium Carbonate on bacterial pathogens “Concentration of 'Na,CO, Streptococcus 50 | 0st 0.06% 0.107 0077 oan 008 0319 0.156 [0s 3sPrevalence of Microbial Inhabitants from Cowshed and ts Coo x Table 2, Effet of Sodium Carbonate on fungal pathouens dis Concentration of | Growth rate of fangal pathogens i mm Fsemicectum flavus t © OD Value (absorbance) at 345 nm Streptococcus sp = 00 Value (absorbance) at Control 36a Research Jounal of Biological Science Inioresence Foret 1, August 2008, p. 33-37 Fig. 2. Effect of Sodium Carbonate on fungal pathogens e — | [© Growth rate of tngal pathogens in mm. F semicectum = Growin rate of fungal pathogens in mm A flaws 1% 2% References ‘Anonymous. (2003), Microbial inhabitants in cow dun, ‘Meenakshi Maruthuva Malar, Meenakshi Mission publishers, Madurai, pp.100. Davis HL. (1987). Temperate pastures: their production use and management, CSIRO, Australia, pp. 446. Francis § M, (1988). Fungus guns 2, Indian Journal of Microbiology, 12:78-79. Ing B. (1989). Why not look at the fungi? Carolina biological supply company, Carolina, a Kendrick Ft K. (1995), The 5 kingdom. Mycological publications, Waterloo, Ontari Muhammed $ and Anusha N/A. (2003), In viteo inhibition 1 Might inducing pattiogens by of growth of some feed compost inhabiting microbes, African Journal of Biotechnology. W61-164 Mondkar D. (1981). Agnihotra and microbes, Scientific data, Maharashira