Bacteriological analysis of vegetables found in local market of Kawardha

Introduction

Vegetables are a vital part of a healthy diet, but they can also harbor a variety of bacteria. Some of these bacteria are harmless, while others can cause foodborne illness if not properly controlled. The fresh vegetables are most recommended and widely consumed highly nutritious form of food. Chemical composition and nutritive value of these product are often influenced by a number of pathogenic microorganisms (Shobha S 2014) [13] . Bacteriological analysis of vegetables is essential for ensuring food safety. By analyzing the bacterial profile of vegetables, we can gain valuable insights into potential food safety risks and implement appropriate measures to minimize them. The primary sources of contamination of microorganisms in fresh vegetables are municipal waste water discharged for irrigation and washing purposes used by local farmers. Village farmers have been found to use river waste water, primarily contaminated by coli form groups, for vegetable washing and irrigation. This practice is known to be the primary source of pathogenic microorganisms in fresh products (Kasture N. 2017) [7] . These vegetables are contaminated during harvesting, transporting and selling in markets (Ekinci M, et al., 2014) [4] . Microbial attacks, pests and climatic changes. The term "food spoiling" describes a variety of alterations that cause food to lose its flavor, texture, aroma, or appearance. Poor handling procedures both before and after harvest were the cause of the vegetable contamination (Yafetto et al., 2019) [18] . The kinds of microorganisms that cause agricultural crops to deteriorate are indicative of a number of microbial problems in those crops (Kaur, A. 2017) [8] . Hand picking, planting and other pre-and post-harvest activities bring them into contact with the veggies. Additionally, there are frequent in-person interactions between vendors and customers in the market where these vegetables are sold (Chaturvedi et al., 2013) [1] . The outermost layers of raw vegetables are contaminated with a variety of microorganisms, depending on a number of factors, including the microbial population of the environment from which the food was taken, the state of the raw product, handling techniques, storage conditions, and time.

The inside tissues of healthy plants have no presence of microorganisms. Fresh vegetables are almost always suggested without quantitative restrictions in diets. Some of these same technologies have also raised the possibility of human sickness due to a variety of dangerous microorganisms. The primary sources of contamination of fresh vegetables are contaminated water, fecal debris, unsanitary conditions, and vendor handling of vegetables and vegetable products. The use of raw animal manure for fertilizer can increase the threat of contamination of vegetables (Sahile S. et al., 2019) [11] . Vegetables are consumed without any heat treatment, sometimes without washing and peeling and therefore the possibility of foodborne diseases is more. Pathogenic bacteria can contaminate vegetables when they are growing in fields or farms (Tambekar et al., 2006) [14] Vegetables can become contaminated by microbes through interaction with processing equipment and the raw materials. The microorganisms that exist on the surfaces of raw, whole produce appear to be the major source of microbial contamination and consequent spoilage of fresh-cut vegetables. If fresh-cut produce is processed in a facility that has been used for produce processing for a long time, it is also possible for people or equipment to come into contact with spoilage bacteria during the processing and packaging stages (Garg et al., 1990). Pathogenic bacteria are present in salad vegetables. (Uwamere O. 2013) [16] . The vegetable market is known for being both heavily trafficked and enriched with human activity. People who work in these regions handle a range of vegetables that come from different places, which exposes them to a lot of organic dust that contains both vegetable debris and a variety of aerosolized microorganisms (Pathak et al., 2009) [9] . Contamination of fresh product with microbial pathogens and antibiotic-resistant bacteria is special concern, because it is likely consumed raw, without any type of cleansing or processing (Hassan et al., 2011) [6] . In order to prevent or minimize bacterial contamination to the lowest level, this study advises everyone participating in the vegetable value chain to enhance their hygiene practices, particularly when handling or processing vegetables at most. The government ought to develop transportable water irrigation methods for growing vegetables and inform growers, retailers, and consumers on the risks posed by these pathogens so that greater care is taken from planting to harvesting. In addition, customers are encouraged to wash veggies well before consuming them.

Materials and Methods

Study site

The study was carried out in local market of Kawardha i.e. vegetable market Kawardha, Naveen market Kawardha, vegetable market Raveli. In the study of present research work three local market areas were selected.

Collection of Samples

Vegetables samples were collected from the local market of Kawardha Chattisgarh. Six samples were collected from the market, which were, Spinach (Spinacia oleracea), Bitter guard (Momordica charantia), Cauliflower (Brassica oleracea var botrytis), Cabbage (Brassica oleracea var. capitata), Brinjal (Solanum melongena), and lady's finger (Abelmoschus esculentus). According to (Dr Ko Tin) sterile polythene zip bags were used for collection of samples. Collected samples were then transported in microbiology laboratory of APSGNS Govt P.G. College Kawardha. The samples were processed for bacteriological analysis within 1-4 hours (Weldezgina et al., 2016) [17] .

Sample Preparation and Dilution

1 Gram (1g) of each sample were weighted then homogenized and added on peptone water. The respective samples were then transferred for serial dilution. Six fold of serial dilution were done using 1 ml of homogenate and 9 ml of sterile distilled water in five test tubes. In sterile petri dish filled with nutrient agar, and 10 -6 dilution were spread out.

Bacterial Count

A volume of 0.1 ml appropriate dilution was spread-plated in duplicate on pre dried surfaces of Plate Count Agar plates. Inoculated plates were incubated at 30 0 c for 24 to 48 hours. For microbial counts, plates with colonies between 30 and 300 were considered. Microbial load in each vegetable sample was determined as CFU/ml and was calculated using formula:-(Kaur, A. and Bhowate, P. 2017) [8] .

CFU = Number of colonies appearing on the plate X Dilution factor Volume of sample taken

Morphological and biochemical Characterization

The isolated strains from vegetables were characterized on basis of morphological and biochemical characterization i. e. Gram Staining, Fermentation of Sugars (Glucose, Lactose and Mannitol), Indole, Methyl red, Voges Proskauer, Citrate, Catalase, Urease and Starch hydrolysis tests.

Antibacterial Susceptibility/Resistance Test

The antibacterial susceptibility/ resistance testing for isolated bacteria were determined according to modified Kirby Bauer disc diffusion technique as described by Clinical Laboratory Standard Institute. The following 3 drugs, namely, Penicillin G, Tetracycline (Sahile, S. et al., 2019) [11] and amoxicillin were used to determine the antibacterial activity of the isolates.

Results and Discussion

Bacteriological analysis of vegetables from local markets is a critical component of ensuring food safety and preventing the spread of food borne illnesses. This analysis involves examining samples of various vegetables for the presence of harmful bacteria, such as Salmonella spp., Pseudomonas spp., Bacillus spp., Citrobacter spp., Staphylococcus spp., and Klebsiella spp., which can pose serious health risks if consumed. A total of fourteen bacterial stains were isolated from spinach (Spinacia oleracea), bitter guard (Momordica charantia), cauliflower (Brassica oleracea var botrytis), cabbage (Brassica oleracea var. capitata), brinjal (Solanum melongena) and lady's finger (Abelmoschus esculentus). In the present study, five gram-positive and nine gram-negative bacteria were isolated. On the basis of morphological and biochemical test three strains, namely V1A, V1C, and V2A, were isolated and identified as Staphylococcus spp. Also detected the presence of Staphylococcus spp. in vegetables. While V1B, V3A, V3B, and V5B strains were identified as Pseudomans spp. Reported the presence of Psedomonas spp. in vegetables. V2C and V5B strains were identified as Bacillus spp. Kaur A. et al. (2017) [8] detect the presence of Bacillus spp. V4B, V5B, and V6A strains were identified as Salmonella spp. Chukwuma OP. (2016) [2] also reported the presence of Salmonella spp., Staphylococcus spp., and Bacillus spp. in vegetables. Pathak et al. (2009) [9] found the presence of Bacillus spp. and Citrobacter spp. and Klebsiella spp. V2B strain is identified as Citrobacter spp., and V6B strain is identified as Klebsiella spp. Several other workers have also detected the same results for bacterial isolates from vegetables. Sahile, S. (2019) [11] reported Klebsiella spp., Psedomonas spp., Staphylococcus spp., and salmonella spp. in vegetables. Dr. Ko Tin reported the presence of Salmonella spp., E. coli, and Staphylococcus spp., Razzaq et al., (2014) [10] found Klebsella spp. in vegetables. E. coli was not isolated from any of the samples tested in the current study. The presence of E. coli in some green leafy vegetables examined. E. coli was not found in any of the samples examined investigation. The antibiotic resistance patterns of Staphylococcus spp., and Salmonella spp. isolated in the current study showed a low percentage of resistance to Penicillin G, Tetracycline, and Amoxicillin. This finding is partly similar to the previous report. Almost 99.99 percentages of Salmonella spp. stains were resistant to all the antibiotics tested in the present study. In the present study, a high number of Pseudomonas spp. was resistant to penicillin G and to tetracycline and amoxicillin.

Conclusion

Bacteriological analysis of vegetables is crucial for ensuring food safety and preventing the spread of foodborne illnesses. In this examination, the number and kinds of bacteria, both good and bad strains, that were present on the surface of vegetables were evaluated.

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