Veterinary Research Forum

Abstract An emerging global threat is the development of resistance to various antibiotic groups among pathogenic bacteria, including Pasteurella multocida. The excessive use of antibiotics and presence of antibiotic resistance carriers play a significant role in the development of multiple resistances to anti-microbial agents. Given the role and importance of plasmids in the development of antibiotic resistance, it is important to prevent the transfer, replication, and multiplication of plasmids by eliminating plasmids. In this study, the role of plasmids in the development and transmission of antibiotic resistance in P. multocida was investigated and evaluated. This study was performed on P. multocida isolates collected from apparently healthy and ailing animals (February to September 2014). Identification of P. multocida isolates was carried out using standard biochemical tests and kmt1 gene amplification. After initial tests and plasmid DNA extraction, five plasmid-containing isolates were selected for further work. To compare antibiotic resistance after plasmid removal, the disk diffusion method was used. Based on the results, it was determined that all plasmid-containing isolates that showed a high level of antibiotic resistance (especially with large plasmids) became sensitive to anti-microbial agents after plasmid removal. Therefore, according to the findings, the role and importance of plasmids in the development of antibiotic resistance in P. multocida was determined. It was also found in this study that the existence and persistence of plasmids by P. multocida bacteria was of great importance in the role of plasmids in the development of antibiotic resistance.

Abstract Bovine clinical mastitis is an economically important disease in dairy industry worldwide resulting in reduction of milk yield and quality. Among mycotic mastitis, Candida spp. are commonly occurring opportunistic mycosis in immunocompromised animals. The micro-organism’s causing mastitis has high zoonotic potential and has been linked with rapid growth and introduction of antimicrobial resistance between animals and humans. The present study was conducted to isolate and identify the common pathogenic Candida spp. from bovine mastitis cases in India. The isolates were phenotypically characterized by culturing on Sabouraud’s dextrose agar, Hichrome Candida differential agar and germ tube production test. Antibiogram was also performed to determine their antifungal activities. The phenotypically positive isolates were confirmed by polymerase chain reaction (PCR) and genetically analyzed by targeting 18S-ITS1-5.8S-ITS2-28S region specific for Candida spp. and identified the yeast at the species level. The antibiogram showed the isolates were highly sensitive with ketoconazole, clotrimazole and miconazole. The PCR assay identified C. lusitaniae and C. tropicalis based on the two distinctive amplicon sizes (592bp and 737bp) respectively. Also, the sequence analysis and phylogeny confirmed C. lusitaniae in six sequences and C. tropicalis in one sequence. It is worth noting that in this study, the species identification was consistent among PCR and genetic analysis. Therefore, the PCR based identification system of the fungal species performed in this study could be an efficient and time saving tool for early diagnosis of clinical mastitis in milch animal, which allows prompt control and application of speedy effective treatment.