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Brief Original Article Prevalence of Microorganisms of Hygienic Interest Brief Original Article Prevalence of microorganisms of hygienic interest in an organized abattoir in Mumbai, India Sudhakar Bhandare, Ashish M. Paturkar, Vikas S. Waskar and Ravindra J. Zende Department of Veterinary Public Health, Bombay Veterinary College, Parel, Mumbai-12, India Abstract Background: The magnitude of food-borne illnesses in India is unknown because of lack of surveillance networks. Monitoring the prevalence of food-borne pathogens and indicators of contamination in primary production at abattoirs is imperative for creating a data bank and for effective control of such pathogens before they enter the food chain. Methodology: Microorganisms of hygienic interest were screened for their prevalence at Deonar Abattoir, Mumbai. Swab samples from 96 sheep/goat carcass sites were collected and analyzed for Staphylococcus spp., Bacillaceae, Clostridiaceae and Enterobacteriaceae. 2 Results: Average Staphylococcus aureus and Staphylococcus epidermidis counts were 3.15 ± 0.18 and 3.46 ± 0.17 log10 CFU/cm , 2 respectively. Bacillus cereus, Bacillus subtilis and Clostridium spp. counts were 3.10 ± 0.08, 3.41 ± 0.19 and 0.76 ± 0.06 log10 CFU/cm , 2 respectively. The Escherichia coli count was 3.54 ± 0.06 and the Klebsiella aerogenes count was 3.22 ± 0.22 log10 CFU/cm . Counts for 2 2 Proteus vulgaris and Proteus mirabilis were 3.44 ± 0.14 log10 CFU/cm and 3.71 ± 0.12 log10 CFU/cm , respectively. S. epidermidis had the highest percentage prevalence at (41.6%), followed by K. aerogenes (31.9%), B. subtilis (28.2%) and P. vulgaris (23.6%). Salmonella spp. were not isolated. Conclusions: The data demonstrate high prevalence and diversity of micro flora on carcasses in the primary Indian production facility, which might be attributed to either human handling or improper dressing especially during evisceration process. Appropriate training for personal and production hygiene is essential for workers in Indian meat production facilities. Key words: microorganisms of hygienic interest; sheep/goat carcasses; abattoir; prevalence J Infect Dev Ctries 2010; 4(7):454-458. (Received 3 March 2010 – Accepted 30 March 2010) Copyright © 2010 Bhandare et al. This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Introduction As far as foods of animal origin are concerned, Food-borne diseases are a threat to public health India is the largest milk producer in the world [4]; worldwide. The repercussions are not only health however, in spite of the potential from the large related but also have economic ramifications from the livestock population, meat production is below loss of business over food safety issues. In the United capacity but is building. The total meat processing States of America (USA), 60% of cases requiring capacity in India is over one million tons per annum, hospitalization are caused by the food-borne bacteria of which 40-50 percent is utilized [5]; thus there is [1], but in developing countries related data is not tremendous scope in this sector. Given the present available because of a lack of precise health-care attention from the government and private infrastructure and data management. It is estimated entrepreneurs, the future of the meat industry in India that annually about 1.8 million children in developing looks prosperous; however, much work is needed in countries succumb to death from acute diarrhoeal maintaining quality assurance to prevent food-borne illnesses, with food or water being the major causes diseases. Monitoring the prevalence of of many of these illnesses [2]. Chugh [3] has microorganisms of hygienic interest in primary identified the emerging and reemerging zoonotic, production at abattoirs is imperative for creating a food-borne and waterborne diseases as well as the data bank and for effective control of such pathogens diseases caused by multidrug resistant organisms as before they further enter the food chain. The present the major public health threats in India. However, the study was undertaken at an organized abattoir in magnitude of food-borne illnesses in India is Mumbai city to determine the prevalence of unknown due to the lack of a coordinated microorganisms of hygienic interest. surveillance network. Table 1. Mean bacterial counts (log10 CFU/cm2 ± SD) and percentage prevalence of various food-borne pathogens isolated from sheep/goat 458 455 - carcasses at Deonar Abattoir, Mumbai. Average Sr. Food borne Percentage After Percentage Percentage After flaying After washing Average counts percentage No pathogen prevalence evisceration prevalence prevalence 2010; 4(7):454 2010; prevalence 2 S. aureus 3.09 ± 0.11 13.9 3.24 ± 0.21 29.1 3.11 ± 0.21 13.8 3.15 ± 0.18 18.9 S. 3 ± ± ± ± epidermidis 3.48 0.16 33.3 3.67 0.13 50.0 3.24 0.22 41.6 3.46 0.17 41.6 4 B. cereus 3.17 ± 0.10 11.1 3.04 ± 0.04 8.3 3.08 ± 0.11 11.1 3.10 ± 0.08 10.2 J Infect Dev Ctries Dev Infect J 5 B. subtilis 3.41 ± 0.31 30.5 3.48 ± 0.15 29.1 3.34 ± 0.12 25 3.41 ± 0.19 28.2 Clostridium 6 0.00 ± 0.00 0.00 2.27 ± 0.17 20.8 2.00 ± 0.00 2.7 0.76 ± 0.06 7.8 spp. 7 E. coli 3.55 ± 0.08 11.1 3.95 ± 0.06 20.8 3.11 ± 0.05 8.3 3.54 ± 0.06 13.4 K. 8 ± ± ± ± aerogenes 3.05 0.15 22.2 3.39 0.30 45.8 3.23 0.20 27.7 3.22 0.22 31.9 9 P. vulgaris 3.32 ± 0.15 30.5 3.89 ± 0.18 29.1 3.10 ± 0.09 11.1 3.44 ± 0.14 23.6 11 P. mirabilis 3.69 ± 0.28 22.2 3.96 ± 0.06 8.3 3.49 ± 0.03 5.5 3.71 ± 0.12 12.0 Salmonella 12 0.00 ± 0.00 0.00 0.00 ± 0.00 0.00 0.00 ± 0.00 0.00 0.00 ± 0.00 0.0 spp. Figure 1. crobes in an abattoir in in Mumbai abattoir in an crobes of mi Prevalence - . et al et Bhandare Bhandare Bhandare et al. – Prevalence of microbes in an abattoir in Mumbai J Infect Dev Ctries 2010; 4(7):454-458. Materials and methods anaerobic technique upon Sodium Polymyxin All the media used were dehydrated and Sulphadiazine (SPS) agar incubated at 44C for 24 purchased from HiMedia laboratories, Mumbai, hours. India. Swab samples were collected from sites on Various members of the family Enterobacteriaceae sheep and goat carcasses after various slaughter were isolated and enumerated using MacConkey agar operations (flaying, evisceration and washing) at an with crystal violet. Eosin Methylene Blue agar was organised abattoir (i.e., a modern, mechanised and employed for isolation and identification of E. coli export-oriented abattoir with proper infrastructure showing typical metallic sheen. To isolate Salmonella and skilled manpower to slaughter a large number of spp. swab samples were homogenised in 225 ml animals, which follows the safety specifications buffered peptone water and pre-enriched at 37°C for given by the Meat and Meat Product Order of 1973 24-48 hours; then 1 ml of culture was transferred to 10 issued by the Directorate of Marketing and ml of Selenite Cystine broth (enrichment medium) and Inspection, Government of India) at Deonar, incubated at 44°C for 18 hours. Selective plating was Mumbai. A total of 96 samples were taken at the done on Brilliant Green Sulpha agar and incubated abattoir from six different carcasses on six occasions 43oC for 24 hours. (Table 1) from the neck, shoulder, brisket, loin, flank Two to three characteristic colonies of each and rump after flaying and washing. After bacterium were further subjected to purification, evisceration samples were collected from the brisket identification, and characterization. Microscopic and rib medial, brisket and rib lateral, and loin and examination was conducted on smears stained by flank regions. Carcass sites were sampled by the modified Gram’s staining as described by Beveridge swab technique. Briefly, an area of 100 cm2 was [6]. Characterisation and identification of organisms marked with a sterile frame of 10 cm x 10 cm for was done according to Barrow and Feltham [7] and each site on the carcass. Sterile cotton wool swabs (3 Cheesbrough [8]. All counts were converted to log10 cm long and 1 cm in diameter) held by wooden sticks CFU/cm2 along with standard deviation and percentage moistened with 0.1% peptone were rubbed on the prevalence was calculated. sites continuously for 30 seconds and transferred to a screw-capped test tube containing 10 ml of sterile Results maintenance medium (0.85% NaCl and 0.1% The results obtained are shown in Table 1 and peptone). The screw-capped test tubes were brought Figure 1. The Staphylococcus spp. isolated were to the laboratory in a thermos flask containing ice and Staphylococcus aureus and Staphylococcus processed immediately. epidermidis with their average counts as 3.15 ± 0.18 2 Test tubes containing swabs were shaken on a and 3.46 ± 0.17 log10 CFU/cm , respectively. From vortex mixer for 30 seconds for uniform distribution of the family Bacillaceae, Bacillus cereus and Bacillus microorganisms. Tenfold serial dilution of all the subtilis were isolated with their average counts as 2 samples was prepared using sterile normal saline 3.10 ± 0.08 and 3.41 ± 0.19 log10 CFU/cm , solution (NSS). Then the samples were processed for respectively. The counts of Clostridium spp. from viable counting. family Clostridiaceae were 0.76 ± 0.06 log10 Food-borne organisms were enumerated using CFU/cm2.
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