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Journal of Food Protection, Vol. 50, No. 5, Pages 401-403 (May 1987) Copyright' International Association ot Milk, Food and Environmental Sanitarians

Survey of the Microbiological Quality of Whole, Black Pepper and Turmeric Powder Sold in Retail Shops in Bombay

H. GEETA and P. R. KULKARNI* Downloaded from http://meridian.allenpress.com/jfp/article-pdf/50/5/401/1657362/0362-028x-50_5_401.pdf by guest on 02 October 2021

Food and Fermentation Technology Division, Department of Chemical Technology, University of Bombay, Bombay-400 019, India (Received for publication February 24, 1986)

ABSTRACT zones, viz. East, West, South, North and Central. Three shops from each zone were selected. Three samples of 2 kg of each Microbiological analysis of loosely packed, whole, black spice were collected from each shop. The environmental condi­ pepper and turmeric powder obtained from retail shops in the tions in the vicinity of shops selected in each zone were as city of Bombay revealed that the samples of both spices were follows: for North Zone, shops were situated in residential col­ highly contaminated. Aerobic plate counts of black pepper onies with clean surroundings; for Central Zone, selected shops ranged from 12.1 X 107 to 81.9 x 108 c.f.u. per gram and were situated by the roadside with hectic activity but were in turmeric powder from 4.1 X 107 to 73.6 X 108 c.f.u. per a fairly clean environment; for East Zone, shops selected were g. In both spices, mesophilic sporeformers like Bacillus oc­ curred. Coliforms ranged in counts from 102-103 per g. Fungal situated on the outskirts of the city in moderately clean sur­ counts ranged from 0.6 X 104 to 16 X 105 per g for black roundings; for West Zone, shops selected were in the vicinity pepper and from 0.5 x 103 to 11.1 x 105 per g for turmeric of unhygienic buffalo sheds; shops in the South Zone were un­ powder. Fungal flora included mainly Aspergillus spp. with the hygienic. occurrence of Mucor in some of the samples. No other or­ The retail shop conditions were temperature, ambient (i.e. ganisms were observed in the dilutions plated. The extent of 30-35°C); relative humidity, 55-70%; and period of storage of contamination was slightly greater in pepper than in turmeric, samples, 4-6 months. The different samples of the whole, black although both spices were of a poor quality when compared pepper and turmeric powder were stored in clean, washed and with international standards. oven sterilized glass bottles until used.

Preparation of the sample Ten grams of each sample were weighed under aseptic condi­ India has been one of the major spice producing and tions and added to 90 ml of sterile saline solution. This was exporting countries from early days. Amongst the spices, taken as the 10"' dilution and further serially diluted up to 10"9 black pepper maintains the highest status and accounts (5). One ml of the required dilution was added to 15-20 ml for a little more than one fourth of the total world trade of the molten and cooled medium using the pour plate method. in spices. Turmeric is another major spice, and is used All media used for plating were purchased from M/s. HiMedia for its coloring ability and medicinal values. Often the Lab. Pvt. Ltd., Bombay. To obtain the total aerobic bacterial 7 8 9 top quality spices are exported and the quality of produce count, 10" , 10" and 10" dilutions were plated using Plate Count agar and plates were incubated at 37°C for 48 h. For available for local consumption may be far from satisfac­ 1 2 3 tory. Spices have been reported to be frequently contami­ coliforms, 10" , 10" and 10" dilutions were plated on Violet Red Bile agar medium and plates were incubated at 37°C for nated {6-9,11,15). However, there are hardly any data 24 h. Baird Parker agar was used for Staphylococcus detection available on the microbial quality of spices sold in loose and isolation for which the dilutions used were 10"', 10"2 and packs in retail markets in a city like Bombay where a 10"3. For selective enrichment and isolation of Salmonella and range of quality, catering to various socio-economic Shigella, 10"', 10"2 and 10"3 dilutions were plated on Bismuth groups situated in specific localities, is found. The pre­ sulfite agar and the plates incubated at 37°C for 24 h (5). Dilu­ sent work was therefore undertaken with a view to sur­ tions of 10"2, 10"3 and 10"4 were plated on Nutrient agar and veying the range of quality of two of the major spices plates were incubated in anaerobic jars at 37°C and 55°C for and finding out the relationship, if any, between the area 7 d to obtain the mesophilic and thermophilic anaerobic count. To detect the presence of Clostridium, the same dilutions were of sampling and microbial quality. plated with Clostridial agar and plates were incubated at 37°C or 55°C for 7 d. Yeast and mold count were obtained with MATERIALS AND METHODS potato dextrose agar (pH 5.6 ± 0.2) using 10"3, 10"* and 10"5 dilutions and the plates incubated at room temperature for 5 Sample collection d. For convenience, the city of Bombay was divided into five

JOURNAL OF FOOD PROTECTION, VOL. 50, MAY 1987 402 GEETA AND KULKARNI

TABLE 1. Microbiological analysis of whole black pepper samples collected from shops in the city of Bombay Aerobic Salmonella sllo P plate Coliform Staphylococcal Shigella Clostridial Yeast and hygienic count count count count count mold count conditions Zone per g per g per g per g per g per g 7 Very good North 35 X 10 3.1 X104 7 36 x 10 <10 <10 <10 <10 2.1 X104 42X107 7 Fairly 2. Central 12X10 1.2X104 7 good 15X10 <10 <10 <10 <10 0.6 X 104 16X107 8 2 Fairly 3. East 42 X 10 42X10 16X105 8 good 36xl0 55X102 <10 <10 <10 6.8 X 105 8 2 59X10 43X10 llxlO5 Poor 4. West 74X108 9.8 X 103 4 15X10 Downloaded from http://meridian.allenpress.com/jfp/article-pdf/50/5/401/1657362/0362-028x-50_5_401.pdf by guest on 02 October 2021 63xl08 13xl03 <10 <10 <10 16xl04 59X108 22X103 17X104 Poor 5. South 81x10s 15X103 7.88 X104 63 x 108 11X103 <10 <10 <10 5.6 X104 59X108 17X103 9.9 X104 "Each result is an average of nine analyses.

TABLE 2. Microbiological analysis of turmeric powder samples collected from retail shops in the city of Bombay" Aerobic Salmonella Shop plate Coliform Staphylococcal Shigella Clostridial Yeast and hygienic count count count count count mold count Zone conditions per g per g per g pcrg per g perg 1 Very 18X107 12X102 7.1 XlO4 (North) good 18X 107 9.5 XlO2 <10 <10 <10 4.1 XlO4 18X107 11 XlO2 3.0 XlO4 2 Fairly 4.1 xlO7 7.2 XlO3 1 XlO4 (Central) good 10X107 5.6 x 103 <10 <10 <10 1.3 XlO4 7.2X107 6.5 xlO3 2 XlO4 3 Fairly 31 X 108 HOxlO2 7.4 XlO5 (East) good 47X108 97 XlO2 <10 <10 <10 4.4 XlO5 45X108 94 xlO2 11 X 10s 4 Poor 20X107 8.7 XlO4 (West) 20X107 <10 <10 <10 <10 8.2 X 104 20X107 8.7 XlO4 5 Poor 64X108 0.5 XlO3 (South) 73X108 <10 <10 <10 <10 0.5 XlO3 70 XlO8 lxlO3 "Each result is an average of nine analyses.

Colonies were counted and results per gram of the sample tation showed higher counts than the other, as expected were calculated. Colony characteristics and gram staining of the for the total bacterial population. Most of the organisms various types of bacteria found were determined and further isolated were gram-positive, sporeforming, catalase-posi- identification and confirmation was done as follows. tive Bacillus spp. These were further identified as B. Identification of Bacillus spp. was done using the method of cereus, Bacillus subtilis, Bacillus megaterium, Bacillus Seenappa and Kempton (12). Bacillus cereus was confirmed by selective growth on Bacillus cereus agar. Coliforms were iden­ licheniformis and Bacillus pumilus. As seen from Table tified by biochemical tests (2,14). Fungi were identified on the 1, coliforms were found in samples from three zones out basis of the color of the colonies on potato dextrose agar, stain­ of the five, where environmental conditions were not so ing characteristics using lactophenol cotton blue and the ar­ hygienic. Staphylococcus, Salmonella and Shigella were rangement of the fungal hyphae and spores (2). absent from all the samples analyzed. Coliforms were further identified as Enterobacter aerogenes and Es­ cherichia coli. Anaerobic organisms were not found in RESULTS any of the samples analyzed. Yeasts were also absent. Among the molds present, Aspergillus niger, Aspergillus Table 1 shows the microbial analysis of whole, black flavus and Aspergillus parasiticus were identified in all pepper collected from shops in the city of Bombay. Of the samples while Mucor spp. were found only in sam­ the five zones, three zones with a poorer degree of sani­ ples from zones No. 3, 4 and 5.

JOURNAL OF FOOD PROTECTION, VOL. 50, MAY 1987 MICROBIOLOGY OF PEPPER AND TURMERIC 403

Table 2 shows the microbial analysis of turmeric pow­ hence should be prevented as far as possible. Second, der from shops in the city of Bombay. Here again, the fungal growth can lead to off-flavors, discoloration and aerobic plate count showed an abundance of gram-posi­ changes in the appearance of spices, making the product tive sporeforming, catalase-positive Bacillus spp. which unacceptable in the market. Third, fungi can alter other­ included B. subtilis, B. cereus, B. pumilus, B. lichenifor- wise unfavorable substrates allowing the growth of bac­ mis and Bacillus coagulans. The aerobic plate count was teria. higher for samples from zones 3, 4 and 5 where the The present study points out clearly the need for im­ levels of sanitation in the environment were poorer than proving the harvesting, post-harvest handling and storage in the first two zones. In contrast to pepper samples, col- conditions of spices in India. Faster drying methods, pro­ iforms were found in turmeric samples from those shops cessing, proper packaging and hygienic, dry storage con­ where degrees of sanitation was higher and absent in the ditions, decontamination treatments in the wholesale and other shops which were situated in a more unhygienic retail shops all can lead to a spice of improved microbial environment. Coliforms included E. aerogenes and E. quality. Environmental hygiene has a definite role to play coli. Also, Salmonella, Shigella and anaerobes were not in the microbial quality of spice and hence must be taken Downloaded from http://meridian.allenpress.com/jfp/article-pdf/50/5/401/1657362/0362-028x-50_5_401.pdf by guest on 02 October 2021 found. Fungal identification showed the presence of A. care of to get a spice of improved quality. flavus, A. parasiticus and A. niger in all the shops, while Mucor spp. were found in all the shops except one. ACKNOWLEDGMENT

DISCUSSION Financial assistance provided by the Department of Atomic Energy, Government of India, is gratefully acknowledged. All the samples of both whole, black pepper and tur­ meric powder contained a heavy microbial load as com­ REFERENCES pared to the international standard specifications (for total count, <106/g; for fungal count, <104/g; for coliform 1. Buchanan, R. E., and W. E. Gibbons. 1975. p. 530. In Bergey's manual of determinative bacteriology, 8th ed. The Williams and count, <10/g of the sample) for these spices. These re­ Wilkins Co., Baltimore. sults are in agreement with reports by other workers 2. Cruickshank, R. 1972. Medical microbiology. Churchill (3,4,6-9,11,15). The samples obtained from the zones Livingstone, Edinburgh, pp. 52, 428-439. No. I and 2, where the environmental conditions were 3. Furia, E. 1977. Pepper - chemistry, microbiology and technology. much better, showed a lower bacterial count for both CRC Rev. Food Sci. Nutr., vol. 9, issue 2. CRC Press, Boca pepper and turmeric. Therefore, environment has a defi­ Raton, FL. 4. Govindrajan, V. S. 1980. Turmeric - chemistry, technology and nite role to play in the microbial load of the spices. quality. CRC Rev. Food Sci. Nutr. 12:212-216. Although in pepper, presence of coliforms was marked 5. Guarino, P. A., and H. J. Peppier. 1976. Spices and condiments. in the unhygienic shops and they were absent in the pp. 568-573. In M. L. Speck (ed.) Compendium of methods for others, turmeric samples showed otherwise. This indi­ microbiological examination of foods. Amer. Public Health Assoc, Washington, D.C. cates unhygienic handling of the spices before and during 6. Julseth, R. M., and R. H. Deibel. 1974. Microbial profile of their storage. selected spices and herbs at import. J. Milk Food Technol. 37:414- Fungal counts did not show any definite correlation 419. with the environment, which is as expected, since fungal 7. Krishnaswamy, M. A., J. D. Patel, K. K. S. Nair, and M. Muthu. contamination occurs before the spices reach the retail 1974. Microbiological quality of certain spices. Indian Spices. shops and the fungal spores remain dormant until favora­ ll(l-2):6. ble conditions for growth are provided. 8. Krishnaswamy, M. A., J. D. Patel, and N. Parthasarathy. 1971. Enumeraiton of micro-organisms in spices and spice mixtures. J. The absence of pathogens like Salmonella, Shigella Food Sci. Technol. 8:191. and Staphylococcus is quite encouraging. Their absence 9. Krishnaswamy, M. A., J. D. Patel, and N. Parthasarathy. 1973. could be due to the antimicrobial effects of spices (13) Some types of colifoms, aerobic mesophilic sporeformers, yeasts or the inhibitory action of some dominant flora over the and molds present in spices. J. Plantatn. Crops (India) I(supple- others (1). ment):200. 10. Maunders, D. T. 1976. Thermophilic flat sour sporeformers. pp. Some reports have shown the occurrence of 254-55. In M. L. Speck (ed.), Compendium of methods for micro­ Staphylococcus aureus (3) and Clostridium in black pep­ biological examination of foods. Amer. Public Health Assoc, per (9). Interestingly, the present work showed the ab­ Washington, D.C. sence of these pathogens in both spices. 11. Powers, E. M., R. Lawyer, and Y. Musouka. 1975. Microbiology The presence of sporeformers, viz. Bacillus and fungi, of processed spices. J. Milk Food Technol. 38:683. suggests the need for proper post-harvest treatment of 12. Seenappa, M., and A. G. Kempton. 1981. A simple key for the identification of Bacillus spp. common in foods. J. Food Sci. Tech­ spices. Although Bacillus spp. in general, do not affect nol. 26:131. public health, B. cereus can lead to food poisoning while 13. Shelef, L. A. 1983. Antimicrobial effect of spices: J. Food Safety B. coagulans leads to flat-sour spoilage (10) and hence 6:29-44. a deterioration of the taste in foods. Presence of E. coli 14. Williams and Wilkins. 1975. Bergey's manual of determinative indicates faecal contamination and hence the possibility bacteriology, 8th ed. Reprinted, Williams and Wilkins Co., Balti­ of enteric pathogens. Fungal spp. like Aspergillus can more. 15. Yessair, J., and O. B. Williams. 1942. Spice contamination and produce potent and carcinogenic toxins (aflatoxins) and its control. Food Res. 7:118.

JOURNAL OF FOOD PROTECTION, VOL. 50, MAY 1987