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International Journal of Recent Scientific Research Vol. 4, Issue, 6, pp.998 1000, July, 2013 ISSN: 0976-3031

International Journal of Recent Scientific Research

RESEARCH ARTICLE A STUDY ON THE MICROBIOLOGICAL QUALITY OF THE PACKAGED SPROUTS MARKETED IN CHENNAI, TAMILNADU
Sangeetha, K., Madanraj, S and *Rajarajan, S
PG and Research dept of Microbiology & Biotechnology, Presidency College (Aut), Chepauk, Chennai-05 ARTICLE INFO
Article History:
Received 13th, June, 2013 Received in revised form 25th, June, 2013 Accepted 14th, July, 2013 Published online 30th July, 2013

ABSTRACT
Sprouts are highly nutritious food, rich in enzymes. In India, sprouts are mainly prepared at home or produced on small scale and sold in local markets in open condition by small units. The microbiological quality of the packed sprouts is studied less in India and especially in Chennai. Hence the Microorganisms were isolated, identified and characterised from the packed sprouts collected from Chennai, India by the standard morphological and biochemical characterisitics. Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Salmonella paratyphi A, Salmonella typhi and Aspergillus niger were the contaminant pathogenic microorganisms and hence these microorganisms could pose a high risk of causing foodborne diseases among consumers,if consumed raw. Copy Right, IJRSR, 2013, Academic Journals. All rights reserved.

Key words:
Sprouts, microorganisms,local markets,Chennai.

INTRODUCTION
Sprouting is the practice of germinating seeds to be eaten either raw or cooked. The consumption of sprouts has increased due to their high nutritional quality, fiber, vitamins, mineral content and low calorific value. Sprouts are made from Pulses (pea family): alfalfa, fenugreek, mung bean, lentil, pea, chickpea, soybean. Cereals: wheat, maize (corn), rice, barley, rye. Oilseeds: sesame, sunflower, almond, hazelnut, linseed.Vegetables and herbs: broccoli, carrot, spinach, cabbage, celery, fennel, onion, parsley, radish, turnip, leek, watercress, mustard, rocket (arugula), lemon grass, lettuce, clover, mizuna, milk thistle, tatsoi. In India, sprouting involves soaking of seeds for 12 hr and sprouting for 24 hr. traditionally, the sprouts were eaten after cooking; however, recently the consumption of uncooked sprouts in salads has increased due to the high nutritious content. In India, the sprouts are mainly prepared at home or produced on small scale and sold in local markets and the shopping malls in big cities have started selling packaged sprouts in refrigerated condition. A variety of pathogenic bacteria, such as Salmonella serotypes (Jerngklinchan and Saitanu 1993; Joce et al.,1990;Mahon et al., 1997;Ponka et al., 1995;Van Beneden 1999) ,E. coli O157:H7(Hara-Kudo et al., 1997, Itoh et al., 1998), Bacillus cereus (Harmon et al., 1987), Aeromonas hydrophila, and Staphylococcus aureus have been isolated from sprouts (Barak, Whitehand, and Charkowski .,2002). Studies have shown attachment and growth of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in alfalfa sprouts (Schoeller, Ingham, and. Ingham. 2002; Ferguson; Lisa 2004) . Between 1995 and 2003, there were 13 outbreaks of Salmonella and E. coli O157:H7 infections associated with commercial sprouts in California (Janet C.Mohle-Boetani et

al., 2001). In 11 of the 13 outbreaks, the pathogen was found to be Salmonella. Salmonella is considered a zero-tolerance pathogen in sprouts. L. monocytogenes has been isolated from sprouts, but no outbreaks related to it have been reported. Studies have shown association of L. monocytogenes with root hairs of alfalfa sprouts and has been reported that washing treatment was not sufficient to remove them. Salmonellosis is the common infection associated with the outbreaks due to eating of raw sprouts (Centers for Disease Control and Prevention. 2002) In India, the sprouts marketed in Mumbai and its suburbs were found to have poor microbiological quality with a high bacterial load and a high percentage of samples were found to be contaminated with pathogens such as Salmonella species, E. coli and coagulase-positive S. aureus (Sunil D.Saroj et al., 2006). Nonpathogenic E. coli was detected in 13% of the mung, 26% of the matki, 40% of the chana, and 19% of the vatana samples. Salmonella typhimurium was detected in 21% of the mung, 40% of the matki, and 4% of the chana samples. Salmonella Dublin was detected in 2% of the mung samples, and Salmonella Washington was detected in 4% of the matki samples. Listeria monocytogenes and E. coli O157:H7 was not detected in any of the samples examined. Coagulase-positive S. aureus was detected in 4% of the mung, 11% of the matki, and 4% of the chana samples. (Sunil Saroj et al., 2006); (Vandan nagar and Jayant Bandekar 2009); (Viswanathan., and R. Kaur. 2001). As there is no other way to guarantee pathogen-free raw sprouts, the U.S. government issued a warning in 1999 regarding the hazard of eating raw sprouts. In response to the increasing number of food borne illnesses, governments all over the world are intensifying their efforts to improve food safety. The World Health Assembly adopted a resolution (WHA 53.15) in which, the World Health Organization (WHO) was asked to give greater emphasis on

* Corresponding author: Rajarajan, S PG and Research dept of Microbiology & Biotechnology, Presidency college(Aut), Chepauk,Chennai-05

International Journal of Recent Scientific Research, Vol. 4, Issue, 7, pp. 998 - 1000, July, 2013 food safetywith the goal of developing suitable, integrated food safety systems for the reduction in health risk along the entire food chain, from primary producer to the consumers.(Sudershan, Pratima Rao and Kalpagam Polasa 2009). Although no serious outbreaks have been reported in India by the consumption of raw sprouts except a few (unpublished data) but a billion of people are affected by salmonellosis each year. There is a lack of study in the microbiological quality of sprouts marketed in the Chennai cities other than Mumbai .Hence the study was carried to identify the pathogenic microorganisms from the raw packed sprouts marketed in Chennai. sprouting has to be followed for each sprout varieties to increase the safety of the product. Though the label claims that the Sprouts were processed in UV treated samples for three samples (BRAND A), it had considerable microorganisms and the microorganisms present in it include Pseudomonas aeruginosa,E.coli,Klebsiella spp;. About 54% of the collected samples showed the growth of Pseudomonas aeruginosa .Though Pseudomonas aeruginosa is not a diarrhoegenic bacteria, it causes diarrhoea in immunocompromised patients. Hence it may pose a risk of causing diarrhoea to the immunocompromised patients,if they consumes it raw. Also the presence of Pseudomonas poses a high risk due to the quorum sensing and biofilm formation by the organism that could result in the transfer of virulence to other bacteria. Escherichia coli isolated in our study had the characteristic biochemical reactions but did not produce metallic sheen showing a typical characteristic of the E.coli O157:H7 which was the causative agent for the outbreak associated with the many sprouts. All the varieties had the growth of Aspergillus niger, a common seed contaminant. Hence the study clearly showed that the seeds were not treated properly which is in accordance with other and earlier reports of Jed W Fahey et al., 2006. The presence of Salmonella paratyphi A and salmonella typhi on the sprouts poses a high risk of succumbing to typhoid fever to the general population. Except the studies done by Sunil Saraj et al 2006 and Nagar and Bandekar, 2009 on the microbiological quality of packed sprouts which had shown to

MATERIALS AND METHODS

Sprout samples of three different brands that were commonly sold were obtained from local market in Chennai (Table 1). About 1 g of each sample was homogenised in 100 ml of sterile physiological saline. After appropriate serial dilutions, 0.1 ml of the serially diluted sample was plated onto the Nutrient agar plate and SDA agar plate for isolation of bacteria and fungi respectively .The sample was spread uniformly using a L-rod. Nutrient agar plates and Sabourauds dextrose agar plates were then incubated at 37C and 28 C respectively. Also the Sprout samples were directly placed on the Nutrient agar plate and Sabourauds dextrose agar plate exposing the cotyledons, hypocotyls for direct contact with the medium for the isolation of bacteria and fungi respectively. The bacterial and fungal growths were identified as per the standard morphological and biochemical tests.

Table 1 Different types of samples collected from different brands

Sample no 1 2 3 4 5 6 7 8 9 10 11 Brand name Brand A Brand A Brand A Brand B Brand B Brand B Brand B Brand B Brand A Brand C Brand C Name of the Sprout Mung bean Sorghum bicolor (White-seeded) Mixed sprouts Black gram Fenugreek Double beans cowpeas Chick pea Horse gram Mung Bean Mixed Sprouts Other specification on the label Cultivated in U.V treated water Cultivated in U.V treated water Cultivated in U.V treated water -

RESULTS AND DISCUSSION

Microorganisms isolated from the different sprouts include Pseudomonas aeruginosa,Escherichia coli,Klebsiella species,Salmonella paratyphi A and Salmonella typhi and Aspergillus niger (Table II). Based on the morphological ,physiological and biochemical characteristics, Pseudomonas aeruginosa, E.coli, Staphylococcus aureus, Salmonella paratyphi A and Salmonella typhi were the clinically important pathogenic microorganisms isolated from different sprouts marketed in Chennai. Aspergillus niger was the only fungi isolated from all the tested sprout samples. In India no reports exist regarding the outbreaks due to sprouts (except unpublished data) and the microorganisms isolated from sprouts other than the studies in Mumbai to the best of our knowledge. Thus this is the first report clearly showing the distribution of microorganisms with different kind of sprouts and hence different treatment procedure of the seeds and

have poor microbiological quality with a high bacterial load contaminated with pathogens such as Salmonella spp, E.coli and coagualase positive S.aureus .Our study provides the first hand report on the microbiological quality and the distribution of pathogenic microorganisms on different sprouts, marketed in Chennai. Food safety has emerged as an important global issue with international trade and public health implications. In response to the increasing number of food borne illnesses, governments all over the world are intensifying their efforts to improve food safety. Owing to the high nutritious value of the raw sprouts, people inclusive of diabetic patients and immunocompromised patients are consuming the sprouts in their diet without cooking. Hence to avoid the gastrointestinal infections, this study recommends the people to eat the cooked sprouts and also the current treatment procedure of the sprouts adopted by the both small scale and large scale industries has to be changed before packaging and marketing to the public to ensure consumers safety.

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International Journal of Recent Scientific Research, Vol. 4, Issue, 7, pp. 998 - 1000, July, 2013 Table 2 Microorganisms isolated from different sprouts
Sample no 1 2 3 4 5 6 7 8 9 10 11 Name of the Sprout Mung bean Sorghum bicolour (White-seeded) Mixed sprouts Black gram Fenugreek Double beans cowpeas Chick pea Horse gram Mung Bean Mixed Sprouts Bacteria isolated Pseudomonas aeruginosa Escherichia coli , Pseudomonas aeruginosa Pseudomonas aeruginosa Klebsiella species Klebsiella species, pseudomonas aeruginosa Staphylooccus species Salmonella paratyphiA Salmonella paratyphiA Salmonella paratyphiA Salmonella paratyphiA Pseudomonas aeruginosa, Salmonella typhi Pseudomonas aeruginosa Klebsiella species, Salmonella typhi Fungi isolated Aspergillus niger Aspergillus niger Aspergillus niger Aspergillus niger Aspergillus niger Aspergillus niger Aspergillus niger Aspergillus niger Aspergillus niger Aspergillus niger Aspergillus niger

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