Browsing by Author "Shathish Sharma, BJ"
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ArticleItem Open Access Detection and Molecular Characterization of Food Borne Pathogens from Ready to Eat Meat and Milk Products(2020-05) Rathnapraba, S; Shathish Sharma, BJ; Kanagaraju, P; Meignanalakshmi, S; Manoharan, S; TANUVASFoodborne illness still remains a major threat to public health. Among the various pathogens causing food borne outbreaks, Salmonella sps, Stapylococcus aureus, Bacillus cereus, Campylobacter sps have been incriminated for majority of outbreak cases. Now – a – days consumption of minimally processed ready to eat (RTE) food products increases the risk of food borne diseases. The aim of the present investigation was to detect the presence of FSSAI notified six food pathogens from ready to eat food products. A total of 250 samples (including milk, milk products, meat and meat products) were screened for the presence of food-borne bacterial pathogens like Staphylococcus aureus, Bacillus cereus, Campylobacter jejuni, Streptococcus sp and Salmonella sp on selective medium. Further, culture positive colonies were screened by colony morphology, gram staining, biochemical analysis and molecular methods. Antibiotic susceptibility test was performed for all isolates of Staphylococcus aureus, Bacillus cereus and Streptococcus spp. The results revealed that the Staphylococcus aureus had 100% resistant against Vancomycin, Methicillin and Penicillin antibiotics and Bacillus cereus isolates had 100% resistant against Vancomycin and Penicillin antibiotics and 100% sensitive towards Methicillin. Whereas the Streptococcus sp. shows 100% resistant against Gentamicin, Streptomycin, Tetracycline and Ciprofloxacin. The prevalence of multidrug resistant pathogenic isolates in ready to eat food products may act as potential source of foodborne intoxication under favourable conditions which need to be addressed.PresentationItem Open Access Detection and molecular characterization of food borne pathogens from ready to eat meat and milk products(2020-02) Shathish Sharma, BJ; Saranya, S; Rathnapraba, S; Meignanalakshmi, S; et al.; TANUVASFoodborne illness still remains a major threat to public health. Among the thirty different pathogens causing food borne outbreaks, Salmonella sps, Stapylococcus aureus, Bacillus cereus, Campylobacter sps have been incriminated for large majority of outbreak cases. Now – a – days consumption of minimally processed ready to eat (RTE) food products increases the risk of food borne diseases. The aim of the present investigation was to detect the presence of FSSAI notified six food pathogens from ready to eat food products. A total of 250 samples (including milk, milk products, meat and meat products) were screened for the presence of food-borne bacterial pathogens like Staphylococcus aureus, Bacillus cereus, Campylobacter jejuni, Streptococcus sp and Salmonella sp on selective medium. Further, culture positive colonies were screened by colony morphology, gram staining, biochemical analysis and molecular methods. Phylogenetic analysis was performed for the 13 isolates including 8 isolates of Staphylococcus aureus, 5 isolates of Bacillus cereus and 3 isolates of Streptococcus. A total of 11 Staphylococcus aureus and 5 Bacillus cereus isolates from meat products were sequence analysed and submitted to Genbank. Antibiotic susceptibility test was performed for all isolates of Staphylococcus aureus, Bacillus cereus and Streptococcus spp. The results revealed that the Staphylococcus aureus had 100% resistant against Vancomycin, Methicillin and Penicillin antibiotics andBacillus cereus isolates had 100% resistant against Vancomycin and Penicillin antibiotics and 100% sensitive towards Methicillin. Whereas the Streptococcus sp. shows 100% resistant against Gentamicin, Streptomycin, Tetracycline and Ciprofloxacin. Hence, these results indicate that the ready to eat food products can act as significant source of antibiotic resistance microbial strains.ArticleItem Open Access Molecular Detection of Food Borne Pathogen Bacillus cereus in Ready to Eat Meat Products(2020-04) Rathnapraba, S; Shathish Sharma, BJ; Saranya, S; Meignanalakshmi, S; TANUVASBacillus cereus is a causative agent for common foodborne outbreaks. In this study, we investigate the presence of food borne bacterial pathogen, Bacillus cereus in ready to eat meat products by cultural and molecular techniques and their antibiotic resistance profiles. Ready to eat meat products (n = 115) were procured from commercial outlets in and around Chennai. Enumeration and isolation of B. cereus was performed by conventional culture and molecular method targeting 185 bp of haemolysin (HL) gene. The isolates obtained were tested for antibiotic sensitivity against commonly used antibiotics. Out of 115 samples screened, 29 found to be positive with 25.2% incidence rate of Bacillus cereus in meat products. The obtained six isolates of B. cereus had 100% resistance against vancomycin and penicillin. The isolates were sensitive towards methicillin, streptomycin, tetracycline and chloramphenicol antibiotics. Hence, the occurrence of B. cereus in ready to eat meat products with the isolates being resistant to antibiotics may cause a serious public health concern and need to be addressed.ThesisItem Restricted PROPIDIUM MONOAZIDE (PMA) QUANTITATIVE POLYMERASE CHAIN REACTION (q PCR) FOR DETECTION OF LIVE BACTERIA IN FOOD SAMPLES(2018) Shathish Sharma, BJ; Rathnapraba, S; Meignanalakshmi, S; TANUVASThe present study was aimed to investigate the presence of foodborne bacterial pathogens in meat products. A total of 100 meat samples (including raw chicken, chicken nuggets, sausage, Kheema, cutlet, meat balls, pakoda, raw mutton, mutton balls, pork sausage, kheema, khabab, ham bacon, beef khabab, cubes, kheema and sausage) have been collected from different retail outlets in Chennai. The overall occurrence of B. cereus, S. aureus and E. coli in different meat samples were found to be 15%, 20% and 14%respectively. All the presumptive colonies were subjected to molecular screening by targeting the Hemolysin gene, fem A gene and mal B gene in B. cereus, S. aureus and E. coli respectively. Based on the molecular characterization, about 11, 5 and 5 isolates of B. cereus, S. aureus and E. coli were identical in screened meat samples. The isolates from the meat and meat products were subjected to antibiotic susceptibility test and the results revealed that all the isolates showed 100% resistance to Vancomycin and Penicillin. Staphylococcus aureus shows 100% resistance to Methicillin and 68.8% sensitivity to ciprofloxacin. In B. cereus they showed 100% sensitivity to methicillin whereas E. coli showed 100% resistance to ampicillin and 100% sensitivity to ciprofloxacin. PMA-qPCR was developed to quantify the viable foodborne bacterial pathogen in meat products. In the present study 2.5 mM concentration of PMA dye was used, which will help to prevent the amplification of DNA from the dead cells. The developed PMA-qPCR had the detection limit of 1X101.25 CFU/g, 1X100.80CFU/g and 1X 100.4 CFU/g forB. cereus, S.aureus and E. coli respectively. These PMA qPCR results were compared with conventional qPCR. These results indicated that the developed PMA-qPCR was more sensitive than the conventional culture method. The sensitivity, specificity and accuracy of developed PMA-qPCR were calculated using Kappa statistics method. The sensitivity of PMA-qPCR for Staphylococcus aureus, Bacillus cereus and Escherichia coli were found to be 60%, 54.4% and 60% respectively. Specificity of the PMA qPCR was found to be 66.7%, 66.7% and 64.4% for Bacillus cereus Staphylococcus aureus and Escherichia coli respectively. The PMA q-PCR could discriminate among viable and dead bacteria it detect only live bacteria from the food samples.