A STUDY OF BIOAEROSOLS IN INDOOR AIR OF FOOD GODOWNS OF VISAKHAPATNAM, INDIA

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1 A STUDY OF BIOAEROSOLS IN INDOOR AIR OF FOOD GODOWNS OF VISAKHAPATNAM, INDIA M. Kiranmai Reddy* 1, T. Srinivas and K. Aruna Lakshmi 2 1. Department of Chemistry, GIT, GITAM University, Visakhapatnam, Andhra Pradesh (INDIA) 2. Department of Biotechnology, GIT, GITAM University, Visakhapatnam, Andhra Pradesh (INDIA) * mkiranmai_reddy@rediffmail.com Received September 10, 2011 Accepted February 5, 2012 ABSTRACT Bioaerosols which include bacteria, fungi, virus and organic dust present in the air can cause several allergic reactions. A study was conducted in five different areas of food godowns in Visakhapatnam (India) to analyze the composition and intensity of bacterial and fungal ores using gravity Petri-plate method. The bacteria were present in the order of 1.8x10 2 to 5.3x10 6. Fungal ores identified in these areas were 1.7x10 3 to 4.2x10 6. The most dominating ecies are Mucor, Rhizopus, Aergillus, Penicillum, Alternaria, Cladoorium, Trichoderma, Fusarium, Pseudomonas, Proteus vulgari and Enterobacter aerogenes. Strains of Aergillus producing as high as µg/kg aflatoxin. Key Words : Bioaerosols, Fungi, Bacteria, Aflatoxin, Visakhapatnam INTRODUCTION There are several cereal storage houses in and around Visakhapatnam for storing rice grains for months. A large quantity of dust is generated in grain storage houses thus causing a potential health risk to workers due to inhalation of grain dust 1. Fungi produce mycotoxin and bacteria endotoxin which can cause allergies to the farm workers 2,3. Improper hygienic conditions may be causes of microbial air pollution 4. Earlier reports, mostly from abroad, revealed that grain dust exposure caused both acute and chronic reiratory impairments 5. It was also reported that grain dust fever and asthma are very much common amongst the workers exposed to grain dust 6. However prolonged exposure may induce both cough and utum 7. Chronic inhalation of grain dust has been shown to adversely affect pulmonary health 8. There are a few reports which deal with exposure to air borne aflatoxin through inhalation 2,9 and have been studied mostly in Europe and North American countries and very less in tropical countries. There are very scanty reports regarding the occupational exposure to fungi, bacteria and other organic dust in India *Author for correondence AIMS AND OBJECTIVES The aim of present study was to evaluate the microbiological contamination levels in different cereal storage houses and food godowns and to assess the degree of exposure to aflatoxin in Visakhapatnam area. MATERIAL AND METHODS Sampling of air was performed from August 2009 to February 2011 in different cereal storage houses and food godowns located in Visakhapatnam. Five different areas were selected viz. Kancherlapalem, Poornamarket, Narshimhanagar, Gajuwaka and Pendurti. Air sampling The air samples were taken by the use of Petri plate method. Two samples were collected from each of the five sites twice a day on two days per week basis for a period of 18 months. Plates were exposed to air 3 minute and 5 minutes reectively so that the air borne microflora can be trapped. The Petri-plates were placed at 0ft to 6ft height above the ground. Microbiological examination of the air Petri-plates containing rose bengal chloromophenicol agar were exposed to air to trap any ore that falls or blow into them. After 446

2 exposing the open petri-plate for 3 to 5 min, the plates were closed and incubated at 25 º C for 5 days. A total of 2880 samples were collected from each site. The fungi developed for five days was observed under a microscope after staining with cotton blue. The cultural and morphological characteristic of fungi were identified by the use of pictorial atlas of soil and seed fungi. 17 For isolating bacteria, nutrient agar plates were exposed to air from 3 to 5 minutes and incubated at 37 º C. Similarly for bacterial ores 2880 samples were collected. Bacterial isolates were identified based on the morphological characters like colony colour, texture followed by biochemical tests performed according to the standard protocols 18. Aflatoxin detection of Aergillus flavus and Aergillus parasiticus was done by indirect ELISA method by the authentic authority. International Crop Research Institute Of Semi-Arid Tropics Hyderabad 19. RESULTS AND DISSCUSSION During the 18 months research work, 30 fungal ecies belonging to 19 genera and 7 bacterial ecies were recorded in different cereal storage houses and food godowns of Visakhapatnam. The concentrations of air borne microorganisms were relatively low during summer season ranging from 1.8x10 2 to 2.2x10 3 and 1.3x10 2 to 2.7x10 3 for bacteria and fungi reectively. The bacterial ores in air were high during rainy season varying from 7x10 4 to 5.3x10 6. On the other hand the fungal ores were reported high during winter season from 8.6x10 4 to 4.2x10 6. (Table 3 and Table 4) The most common fungi were Aergillus flavus 92% during winter season in kancherlapalem food godowns and 91% in gajuwaka cereal storage houses. The other fungal ores reported were Aergillus parasiticus ranged from 9% to 62%, Aergillus niger 5% to 57%, Aergillus fumigatus 3% to 39%. Similarly, Alternaria alternata are high during winter season in poornamarket 73% followed by pendurti 72%. Some other ecies like Botrytis, Cephaloorium, Cercoora, Cladoorium, Curvularia, Fusarium, Helminthoorium, Table 1 : Percentage of fungal colonies in food godowns S/N Fungal Species 447

3 Journal of Environmental Research And Development Vol. 6 No. 3, Jan-March 2012 Table 2 : Percentage of bacterial ecies S/N Bacterial Species Table 3 : Bacteria and fungi colony forming units S/N Sampling area 448

4 Table 4 : Identification of bacteria S/N Test Bacillus Staphylococcus Streptococcus Escherichia Enterobacter Proteus Pseudomonas subtilis aureus lactis coli aerogenes vulgaris 1 Gram Staining Rod, G+ Coccus G+ Coccus G+ Rod G- Rod G- Rod G- Rod G- 2 Cultural characteristics Abundant, Abundant Thin, even White, moist of Abundant thick Thin, blue Abundant thin opaque, white, opaque growth glistening white glistening gray, swarming white, medium waxy Golden growth growth turns green 3 Gelatin liquefactions rapid 4 Starch hydrolysis Lipid hydrolysis Litmus milk test peptonization Acid reduction Acid rapid Acid, Curd + gas Acid Alkaline Rapid reduction reduction peptonization 7 Lactose fermentation - A A AG AG Sucrose fermentation A A A A+ AG+ AG+ - 9 Dextrose fermentation A A A AG AG AG - 10 Nitrate reduction Indole production H 2 S production M.R Test V.P Test Citrate utilization Catalase activity Urease activity Oxidase activity

5 Table 5 : Aflatoxin µg / kg S/N Sampling Area Aergillus flavus Aergillus parasiticus 1 Kancherlapalem Poornamarket Narshimhanagar Gajuwaka Pendurti Mortierella, Mucor, Penicillium, Rhizopus, Stachybotrys, Trichoderma, Trichothecium, Verticillium and Yeast cells also varied from season to season as shown in Table 1. Similarly, bacterial ores like Bacillus subtilis varied from 16% to 88% and Enterobacter aerogenes 7% to 75% were high during rainy season. The percentage of Escherichia coli, Proteus vulgaris, Pseudomonas, Streptococcus aureus and Staphylococcus lactis were shown in Table 2. Isolates of A. flavus and A. parasiticus were inoculated on corn for 14 days and then aflatoxin is detected. The aflatoxin produced by A. flavus was as high as µg/kg in kancherlapalem food godowns where as A. parasituxus was µg/kg. Aflatoxin in five different areas was tabulated in Table 5. The different ore concentrations of air borne microorganisms inside the godowns were higher in winter season compared to summer. Our study also reported as high presence of microorganisms in winter season 22. Improper functioning of ventilation systems 20, storage moisture of feed rations, kinds of work practice and climatic conditions determine bioaerosols in air. In our study these conditions were seen. Both bacteria and fungal ore concentration in different godowns were different in summer, winter and rainy season. On an average bacteria like Enterobacter and Pseudomonas. were in the air which produced endotoxin which were considered as potential agents of occupational diseases 7. In the present study the bacteria were present in all the sampling sites posing a potential risk of allergies. Diseases like asthma and reiratory tract allergies were noticed in the workers of food godowns exposed to Aergillus and Alternaria In our study these two genera were present at high levels. Fungal and bacterial ores are more difficult to diagnose and treat than other allergies since fungi are more numerous and antigenically variable than other allergens and are exceedingly difficult to avoid 22,23. The occurrence of micro fungi is related to the presence of allergenic and toxin produced in food godowns Aergillus flavus strains have been recovered from the waste diosal area of the mill 15. In the present work Aergillus flavus and Aergillus parasiticus are observed as producing high aflatoxins in different sampling areas. CONCLUSION To prevent reiratory problems it is essential to control air borne ores either through wearing protective mask or practicing proper personal hygiene. Studies in this line are common in developed countries which resulted in timely health alters to the stakeholders. But developing countries too should turn their attention and promote similar studies and the results be used to provide health alters and safety precautions to the working class and safe guard their health. REFERENCES 1. Williams N, Skoulas A and Merriman J., Exposure to grain dust, A survey of health effects, J. Occup. Med., 6(1) , (1964). 2. Depico G. A., Redden W., Tsiatis A., Peters M.E. and Rankin J., Epidemiological study of clinical and physiological parameters in grain handlers of northern United States, Am. Rev. Reir. Dis., 130(1), , (1964). 3. Chan-Yeung M., Wong R. and Maclean L., Reiratory abnormalities among grain 450

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