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Mastitis is a global disease occurring in dairy cows, causing notable economic losses. Extensive use of antibiotics could allow the emergence of mobile antimicrobial resistance genes in mastitis-causing pathogens. This study aimed to investigate the prevalence and characterization of colistin resistance genes in E. coli recovered from bovine mastitic milk. A total of 74 E. coli isolates were investigated for antimicrobial resistance. The presence of mcr-1, mcr-2, mcr-3, mcr-4, and mcr-5 plasmid-mediated resistance genes, as the most crucial contributors to resistance to colistin, was examined by Multiplex PCR. Antimicrobial susceptibility patterns of all isolates to the seven most common antibiotics applied in dairy herds, including colistin, ceftriaxone, ampicillin, tetracycline, gentamicin, enrofloxacin, and trimethoprim-sulfamethoxazole were determined by the DD test. Among all samples, 70 isolates (94.6%) were resistant to colistin. In the MIC test, all isolates were also resistant to colistin, which was in agreement with the DD test. None of the E. coli isolates carried plasmid-mediated colistin resistance mcr-1 to 5 genes in Multiplex PCR. Despite the important role of food-producing animals in the transfer of antibiotic resistance, mastitis-causing E. coli isolates were not the source of mcr 1 to 5 genes in this study. The present research showed a high level of phenotypic resistance to colistin, while there was no agreement with their genotypic resistance. Consumption of polymyxins in dairy calves and the probable existence of other more effective resistance genes could be the reason for this high rate of phenotypic resistance. [ABSTRACT FROM AUTHOR]

The aim of the present study was to evaluate the efficacy of macrolides to eliminate intramammary infection (IMI) caused by Staphylococcus aureus (S. aureus) and Streptoccus spp. 3 weeks before calving time. Eighty Holstein dairy cows with subclinical mastitis pathogens were divided into three groups. Three weeks before expected parturition time, cows in group 1 received tilmicosin (n = 29), cows in group 2 received tylosin (n = 30) and cows in group 3 were left as negative control (n = 21). Milk samples were obtained on 3 and 7 days after calving. Randomly amplified polymorphic DNA (RAPD) method was determined for all of the S. aureus isolates that had the same isolates before and after parturition. The total cure rate was 63.33, 75.86 and 66.66% for tylosin, tilmicosin and control groups, respectively. Furthermore, cure rates were not significant, when each type of mastitis causing pathogens were considered separately. The incidence of clinical mastitis during 60 days after calving for tylosin, tilmicosin and Control groups was 23.33, 27.58 and 38.09%, respectively. Only four S. aureus isolated before drying-off were similar to post-calving isolate, according to RAPD-PCR method. In conclusion, antibiotic therapy before calving improved the cure rate numerically, however, it was not significant. [ABSTRACT FROM AUTHOR]

The present study was performed to assess the resistance profile to fluoroquinolone and to determine mutations in gyrA and parC genes of Escherichia coli in bovine coliform mastitis. Fluoroquinolones (norfl oxacin (NOR), ciprofl oxacin (CIP), enrofl oxacin (NFX), levofloxacin (LEV), and ofloxacin (OFL) were tested against E. coli isolates, isolated from bovine mastitis (100 milk samples) by disk diff usion method. To determine the extent of gyrA and parC mutations associated with fluoroquinolone resistance in E. coli, two isolates with the highest resistance to each fluoroquinolone were submitted for the amplification and sequencing of the quinolone resistance-determining regions (QRDRs) of gyrA and parC genes. Th e disk diffusion method indicated that E. coli isolates had the highest intermediate resistance to OFL (16.7%), followed by NFX and NOR (15%), while they had low resistance to CIP and LEV (3.33%). A few silent mutations in gyrA (in codons 91, 100, 111, 131, 132) and in parC (in codons 91, 157, 159) were detected in QRDRs, and mutations in nucleotides 65, 80, and 83 in gyrA, and 195, 209, 212 in parC were detected in the other isolate. These results showed an intermediate rate of resistance to fl uoroquinolones in E. coli isolates from raw milk of cows with coliform mastitis [ABSTRACT FROM AUTHOR]

Background and Objectives: Escherichia coli is responsible for various enteric and extraintestinal infections in animals and humans. Iron as an essential nutrient, has a proven role in pathogenicity of E. coli. Pathogenic E. coli benefits of having complicated systems for iron acquisition but our current knowledge is limited because of complexity of these systems. In the present study, three multiplex-PCR assays were developed to screen nine different virulence genes related to diverse iron acquisition systems in E. coli. Materials and Methods: The multiplex-PCR systems were designed and optimized in three panels. Each panel includes a triplex-PCR cocktail. The panels are as follow: panel 1: iroN, iutA and fecA; panel 2: fyuA, sitA and irp2; and panel 3: iucD, chuA and tonB. A total of 39 pathogenic E. coli was screened according to the designed multiplex-PCR. Results: In total, the top three frequent genes were tonB (100%), fecA (66.6%) and sitA (58.9%). With the exception of fecA and tonB, comparing the prevalence of genes among different origin of isolates (human, cattle, poultry and pigeon) showed significant associations (P < 0.05). Moreover, the iroN, sitA and iucD genes were significantly prevalent (P < 0.05) among members of extraintestinal pathogenic E. coli in comparison with the group of diarrheagenic E. coli. Conclusion: The current multiplex-PCR assays could be a valuable, rapid and economic tool to investigate diverse iron acquisition systems in E. coli for more precise virulence typing of pathogenic or commensal strains. [ABSTRACT FROM AUTHOR]