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Not AvailableCampylobacter is one of the important foodborne zoonotic bacterial pathogen that causes enteric disorders in animals, birds as well as in humans. The organism is fastidious in nature, requires microaerophilic environment for its growth and survival. Morphologically, it is gram- negative rods with spiral and gull wing appearance. Polymerase chain reaction (PCR) is gold standard method for the detection of Campylobacter from clinical and food samples. For performing PCR, extraction of DNA to be used as template is a challenging task due to lysis resistant nature of bacteria. The genomic DNA isolation was attempted from pure cultures of Campylobacter by three methods viz. Snap-chill, Salt-Tris EDTA (STE) and Columns based commercial kit method. The average concentration of extracted DNA was highest in STE method (03 -3500 ng/μl) followed by Kit method (03 -2000 ng/μl) and Snap-chill method (00-20 ng/μl). The absorbance ratio at 260 nm and 280 nm (A260/A280) was high up to 1.90 in STE method followed by up to 1.80 with column-based kit and 1.5 with snap-chill method which reflects the high purity of isolated DNA by STE and kit-based protocol. As per the results STE method and Kit method was comparable to each other but due to high cost of commercial kit, STE method is proposed to be desirable and may be used routinely for extraction of DNA of lysis resistant bacteria. The PCR results also advocate the preference of STE method over kit method while Snap-chill method was not found effective for lysis resistant Campylobacter isolates.ICAR Sponsored project-Outreach Programme on Zoonotic Diseases

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Not AvailableCampylobacter species are one of the important foodborne zoonotic bacterial pathogens triggering enteritis in domestic, wild captive and non-captive animals as well as birds that spreads through different environmental sources as water, feed, meat and milk. In the present study comparative isolation and identification of Campylobacter jejuni and Campylobacter coli was attempted from faecal samples of wild animals during the year 2020-2021. A total of 430 faecal samples were collected from zoos/wild life sanctuaries and national parks of Uttar Pradesh, Uttarakhand and Chhattisgarh states. Since thermophilic Campylobacters- target bacteria are microaerophilic and fastidious in nature, require specific temperature and nutrition for its optimum growth, hence to find out the suitable selective media, present study was designed. After aseptic collection, processing and primary isolation, growth of identified Campylobacters were assessed on five artificial media, broadly categorized into blood free and blood containing media. The overall positivity of Campylobacter spp. was 59 of 430 (13.72%) with Campylobacter jejuni (7.67 per cent) and Campylobacter coli (6.05 per cent). After enrichment, plating on CBA with selective supplement yielded a significantly higher (P<0.05) prevalence (4.65%) of Campylobacter species. No significant differences could be observed in mCCDA (3.72%) and BA (3.48%). Least isolation observed on CA (2.0%) and HCCA (1.16%). Multiplex PCR results confirmed speciation as well as sensitivity of each culture methods. As the majority of Campylobacter spp. were isolated by CBA with selective supplement. Hence, this may be the method of choice for isolation of Campylobacter species because of presence of hemin in sheep blood as oxygen quenching agent.ICAR Sponsored project-Outreach Programme on Zoonotic Diseases