Toll Free Helpline (India): 1800 1234 070

Rest of World: +91-9810852116

Free Publication Certificate

Vol. 11, Special Issue 2 (2022)

Isolation, identification, molecular characterization and antibiogram studies of Salmonella spp. isolated from calf diarrhea in and around Udaipur (Rajasthan)

Author(s):
Anju Chaudhary, Sudeep Solanki, Deepak Sharma, Rajesh Singathia, Karishma Rathore, Arpita Sain and Durga Devi
Abstract:
Background: Calf diarrhoea has been a major threat to dairy farms and industry. It is a multi-etiology disease with Salmonella spp.as one of the predominant cause. Emergence of antimicrobial resistance is of serious concern because it has rendered the present day antibiotics less effective. The present study aims for isolation, molecular characterization and antimicrobial resistant profiling of Salmonella spp. from calf.
Method: A total of 100 fecal samples were collected from calves of cattle and buffalo of below 3 months of age. Sterile fecal swabs (Hi-media, India) were used for sample collection. All the samples were transported immediately to the laboratory under cold chain and were processed for isolation of Salmonella spp. using standard microbiological techniques. For isolation and identification of Salmonella spp. a loopful of fecal sample was enriched in Selenite broth and incubated at 42 °C for 12-14 hours. After that a loopful of selenite broth was inoculated and streaked separately onto selective agar plates as xylose Lysine Deoxycholate (XLD) agar, Brilliant Green agar (BGA), MacConkeyꞌs agar and incubated at 37 ℃ for 24hours. Partially identified cultures were suspended in normal saline and smeared over a slide. The smears were allowed to air dry followed by heat fixing and Gram’s staining. All Gram negative culture was subjected to biochemical characterization by using various biochemical tests such as catalase, oxidation, oxidation-fermentation, Indole, methyl –red, Voges-Poskauer, citrate test, TSI and urease test. Molecular characterization by Genomic DNA isolation using heat treatment method All the biochemically confirmed isolates for Salmonella spp. Where subjected to PCR for the presence of invA gene (284bp) genus specific primer (F-GTGAAATTATCGCCACGTTCGGGCAA) (R-TCATCGCACCGTCAAAGGAACC) (Borges et al., 2017) [4] were used to amplify Salmonella strain.
Results: Only 3% (3/100) of them were found positive for invA (universal stress protein) gene. The nucleotide sequences flanking the gene encoding the invA and was found to be highly specific for Salmonella. The antibiogram of Salmonella isolates by Kirby Bauer disc diffusion method revealed highest resistance against azithromycin, streptomycin and tetracycline and lowest against ciprofloxacin and gentamicin. All three isolates were MDR. Hence, surveillance based study and search for an alternative to antibiotics is needed to address the global issue of the antimicrobial resistance.
Pages: 1413-1417  |  407 Views  167 Downloads
How to cite this article:
Anju Chaudhary, Sudeep Solanki, Deepak Sharma, Rajesh Singathia, Karishma Rathore, Arpita Sain and Durga Devi. Isolation, identification, molecular characterization and antibiogram studies of Salmonella spp. isolated from calf diarrhea in and around Udaipur (Rajasthan). The Pharma Innovation Journal. 2022; 11(2S): 1413-1417.

Call for book chapter