Biotyping of Bacillus cereus from the street vended foods in

Aim: The present study was undertaken to describe the biotyping of Bacillus cereus isolated from different street vended mutton tikka and chutney samples. 
 
Materials and Methods: A total of 100 street vended food samples comprising of 60 mutton tikka and 40 chutney samples were tested. 
 
Results: The biotype 3 and biotype 4 showed the highest occurrence with, 29.63% and 25.93% isolates falling in these biotypes, respectively. The percentage occurrence of the biotypes 1, 6, 2, 5, and 7 was 14.81%, 11.11%, 7.40%, 7.40% and 3.84%, respectively. The most common found biotypes in Mutton tikka were biotypes 3(29.63%), 4(25.93%), 1(14.81%) and 6(11.11%). The Bacillus cereus strains isolated from chutney samples could be divided into 7 of the 9 possible biotypes. The biotypes 6 and 7 showed the highest occurrence with 38.46% and 30.76% falling in these biotypes, respectively. The biotype 5 and 2 were prevalent to the extent of 23.07%, 7.69%, respectively. The biotypes 3, 4 and 1 were absent. The mean bacterial count of 60 mutton tikka and 40 chutney samples was 4.6817 and 5.6575 log cfu/g. 10 
 
Conclusion: The field isolates and the standard strains of Bacillus cereus had similar cultural, morphological and biochemical characteristics. The biotypes recovered from the Mutton tikka samples were biotypes 3, 4, 1 and 6 and in chutneys the biotypes recovered were 6, 7, 5 and 2. The strains of Bacillus cereus were highly resistant to penicillin G (92.59%).


Introduction
Street foods are frequently associated with diarrhoeal diseases and occur due to improper use of additives, presence of pathogenic bacteria, environmental contaminants and disregard of good manufacturing practices (GMPs) and good hygienic practices (GHPs).Foodborne bacterial pathogens commonly detected in street-vended foods are Bacillus cereus, [1].Street foods are ready-to-eat foods and Clostridium perfringens, Staphylococcus aureus, Listeria beverages prepared and/or sold by vendors and monocytogenes, Escherchia coli and Salmonella spp.hawkers on the street from pushcarts or baskets or [6,7].Bacillus cereus is widely spread in the environbalance pole, or from stalls or shops having fewer than ment (soil, water and dust) from where it easily four permanent Walls [2].Street vended food types contaminates foods of both plant and animal origin differ greatly between countries and cultures and the such as pulses, vegetables, milk, meat and their products industry plays an important role in meeting the food [8].It possesses a diversified animal pathogencity and requirements of urban dwellers in many cities and produces two distinct types of enterotoxins, one of towns of developing countries [3].However, food which is highly heat stable, withstanding the tempo borne illnesses of microbial origin pose a major health erature of 126 C for 90 minutes.Bacillus cereus is an problem with street vended foods [4,5].
important cause of foodborne illness in man and is Street food vending has become an important public health issue and a great concern to everybody.This is due to widespread food borne diseases, due to the mushrooming of wayside food vendors who lack an adequate understanding of the basic food safety issues frequently involved in a number of foodborne out-Results breaks worldwide [9].
Biotyping: Using the biotyping scheme proposed by Kashmir is famous for its non-vegetarian cuisines [10] the study was extended to the fermentation of and most of these preparations are also sold in the three Ammonium salt sugars viz; xylose, salicin and market as vended foods.Common non-vegetarian street cellubiose.Of the theoretically 9 possible biotypes, the foods of Kashmir include Ristas, Kababs, Kantis, Bacillus cereus isolates from Mutton tikka could be Roganjosh, Fried fish and Mutton tikka, the latter divided into 7 biotypes.The biotype 3 and biotype 4 being most common and widely accepted street showed the highest occurrence with, 29.63% and vended food of Kashmir particularly of Srinagar city.25.93% isolates falling in these biotypes, respectively.Therefore, present study was conducted to know the The percentage occurrence of the biotypes 1, 6, 2, 5, Biotyping of Bacillus cereus from the street vended and 7 was 14.81%, 11.11%, 7.40%, 7.40% and 3.84%, mutton tikka and chutney samples in Srinagar city of respectively (Table -3).The most common found Kashmir valley of Jammu and Kashmir State, India.

Materials and Methods
(25.93%), 1(14.81%) and 6(11.11%)(Table -3).There was not much selectivity in the distribution of biotypes Sampling: Bacteriological quality of 100 street with regard to sampling area of Mutton tikka.vended food samples comprising 60 Mutton tikka However, biotype 7 could not be isolated from any samples and 40 chutney samples collected randomly zone except south zone.Biotype 2 was absent from from vendors of five zones of Srinagar city viz.East, central, north and west zone.Similarly biotype 5 was West, North, South and Central was studied in two absent from north, east and west zone.Biotype 1 and 6 subsequent seasons viz.autumn and winter.For this were absent from east and south zone.purpose, 6 samples of mutton tikka and 4 samples of The Bacillus cereus strains isolated from chutney chutney per zone per season were brought to veterinary samples could be divided into 7 of the 9 possible public health laboratory in ice and processed within 2biotypes.The biotypes 6 and 7 showed the highest 3hr for determination of various Bacillus cereus occurrence with 38.46% and 30.76% falling in these serotypes.The nature and the zone wise sampling biotypes, respectively.The biotype 5 and 2 were pattern are presented in Table-1.
prevalent to the extent of 23.07%, 7.69%, respectively.Biotyping of Bacillus cereus:The isolates were The biotypes 3, 4 and 1 were absent (Table 4).The biotyped based on their ability to ferment ammonium distribution of biotypes from different areas did not salt sugars viz.xylose, salicin and cellubiose as per the reveal any selectivity.scheme described by [10] (Table-2).Attempts were However, biotype 3, 4 and 1 could not be isolated made to locate the source of these biotypes in Mutton from any zone, whereas, biotype 5, 6 and 7 were absent tikka on the basis of their previous history and in north and east zone.Similarly, biotype 2 was absent available literature.
in all zones except central zone.The most common
The scheme to upgrade hygiene and quality of Biotyping has been used a simple tool to study street food has also been undertaken by the Ministry of the epidemiology of Bacillus cereus [10].The scheme Food Processing Industries, India.Under the proposed has been used exclusively for locating the source of programme, 10,000 street food vendors will be contamination of different foods by Bacillus cereus identified, and the majority of stake-holders will be [11,12].The ability of the isolates to ferment three upgraded in terms of quality and hygiene and efforts ammonium salt sugars viz.xylose, salicin and would be made to make it mandatory for the vendors to cellibiose has been used as a basis for the biotyping of register with the local authorities [21].Bacillus cereus.In the present study, the isolates of Bacillus cereus were grouped into 7 of the 9 Author's Contribution theoretically possible biotypes.The biotyping of Yasir Hafeez implement the study design.Asif iqbal isolates from mutton tikka revealed predominance of and Manzoor Ahmad collected the samples, drafted biotypes 3, 4, 1 and 6 with 29.63% isolates typed as the manuscript and revised manuscript.Asif iqbal and biotype 3, 25.93% as biotype 4, 14.81% as biotype 1 Manzoor ahmad also help in collection of samples.and 11.11% as biotype 6. Biotypes 3 and 4 have been Yasir Hafeez and Manzoor Ahmad helped in reported as most prevalent biotypes in meat and meat interpretation of data.All authors read approved the products by [10] and while typing heat resistant final manuscript.Bacillus cereus strains from boiled rice, found biotypes 7, 6, 5 and 1 as the most common biotypes Acknowledgements [12].Predominance of biotype 7, 6, and 5 from fried The authors are thankful to the Dean, Faculty of rice has earlier been reported by [11].Similarly the Veterinary Science and Animal Husbandry, Shuhama, biotyping of isolates from chutney samples revealed Alusteng, Srinagar, for providing the necessary predominance of biotypes 6, 7, 5 and 2 with 38.46% facilities at the time of research.isolates typed as biotype 6, 30.76% as biotype 7, 23.07% as biotype 5, 7.69% as biotype 2. The most Competing interests prevalent biotypes recovered from the mutton tikka Authors declare that they have no competing interests.samples were 3 and 4, which indicates meat as the source of Bacillus cereus in mutton tikka samples.References These findings are in the agreement with the findings of [10].Similarly in the chutney samples the common biotypes were 6 and 7, indicating spices as the source of contamination as was also reported by earlier 2.

Table - 2
. Biotyping scheme as proposed by Jha and biotypes recovered from chutney samples were biotypes

Table - 1
. Zone wise sampling pattern of Mutton tikka and the Chutney supplied with it.
. The mean viable counts recorded in

Table - 3
. The distribution of biotypes in Mutton tikka samples from different zones.

Table - 4
. The distribution of biotypes in Chutney samples from different zones.Biotyping of Bacillus cereus from the street vended foods in Srinagar area of Kashmir, India