Veterinary Research Forum

Abstract Bovine subclinical mastitis represents a major cause of severe economic losses in dairy farms. This research aimed to detect the antimicrobial resistance trends of Staphylococcus aureus and to determine the presence of mecA, mphC, lnuA, tetK and tetL antimicrobial resistance genes in raw bulk milk in the period between December 2023 and February 2024. One hundred raw bulk cow milk samples were gathered from different dairy farms in Egypt. The prevalence of subclinical bovine mastitis was 65.00% using California mastitis test. The prevalence of isolated S. aureus was 46.15% via bacterial culturing and all isolates (n = 30) were confirmed via hemolytic activity, catalase and coagulase test, and gram staining followed by polymerase chain reaction targeting nuc1 gene. Antimicrobial sensitivity test was applied on all confirmed S. aureus isolates utilizing the disk diffusion method on Mueller-Hinton agar. The highest resistance was verified for tetracycline at 100% followed by erythromycin and clindamycin at 56.66 and 16.66%, respectively. The highest sensitivity at 100% was verified for amikacin, ampicillin, amoxicillin plus clavulanic acid, ampicillin plus sulbactam, ciprofloxacin, colistin, gentamicin, imipenem, tobramycin, doxycycline and vancomycin. Multidrug resistance was found in 20.00% of the total isolates. Methicillin resistant S. aureus represented by mecA gene was identified in 83.33% of isolates. Macrolides resis­tant S. aureus represented by mphC gene was identified in 16.66% of isolates. Lincosamide resistant S. aureus represented by inuA gene was identified in 66.66% of isolates. Tetracycline resistant S. aureus represented by tetK and tetL genes was detected in 23.33 and 53.33% of isolates, respectively. This study provided antibiotic-resistant S. aureus profiles to dairy farms to avoid treatment failure, adverse effects on animal health and economic impact for the owner of the animal.

Abstract Staphylococcus aureus is one of the most common causes of mastitis worldwide. This study aimed to determine the prevalence and antimicrobial resistance (AMR) patterns of S. aureus in mastitic milk samples collected from camel farms in Matrouh Governorate, Egypt. A total of 200 mastitic camel milk samples were evaluated for S. aureus using both conventional culture-based and molecular-based methods. Antibiotic susceptibility testing of S. aureus isolates was conducted using disc diffusion and agar dilution methods, with antibiotic resistance genes identified through polymerase chain reaction with specific primers. Out of samples tested, 60 (30.00%) were positive for S. aureus. The isolates displayed the highest of resistance against piperacillin-tazobactam (55.00%) followed by trimethoprim- sulfamethoxazole (45.00%) and amoxicillin (40.00%). Half of the isolates were multidrug-resistant (MDR). The AMR genes included methicillin-resistant gene (mecA), β-lactamase gene (blaZ), tetracycline resistance gene (tetK), erythromycin resistance gene (ermB) and vancomycin resistant gene (vanA) were detected in 100%, 100%, 95.00%, 90.00% and 20.00% of the isolates, respectively. In conclusion, the presence of MDRS aureus as a cause of clinical camel mastitis is a significant veterinary and public health concern. These findings highlight the importance of proper antibiotic use in Egyptian camel farms and the need for molecular techniques to fully understand the genetic profile of antimicrobial-resistant S. aureus isolates.

Abstract Staphylococcus aureus is gaining worldwide attention because of its substantial impact on public health. The current study aimed to characterize S. aureus strains isolated from wild birds in the Kasur district of Punjab, Pakistan from 2021 to 2022. A total of one hundred samples were collected from five wild bird species. The samples were enriched, inoculated on selective agars and cultured for 24 hr at 37.00 ˚C. All isolates were verified by matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF MS) and polymerase chain reaction (PCR) after Gram staining. Positive isolates were screened for phenotypic (Kirby-Bauer disk diffusion and minimum inhibitory concentration s), genotypic antibiotic resistance, and virulence genes. These samples yielded 30 (30.00%) S. aureus isolates, confirmed by polymerase chain reaction utilizing the 16S rRNA gene. Staphylococcus aureus was more prevalent in cloacal samples (16.00%) than oral samples (14.00%). Various S. aureus isolates showed varying degrees of resistance to three different antibiotics. Oxacillin (56.66%; n = 17) and tetracycline (33.33%; n = 10) showed the highest resistance rates with the lowest susceptibility (43.33%; n = 13). In contrast, vancomycin, rifampicin, linezolid, and daptomycin were 100% susceptible. Further disc diffusion study revealed resistance to tetracycline (33.33%), erythromycin (16.66%), and gentamicin (10.00%). The tetK gene was found in 33.33% of wild bird samples, while the ermA gene was found in 16.66% of samples. The aacA-D gene was only found in three (10.00%) isolates. None of the isolates tested positive for virulence genes. In conclusion, S. aureus is carried by wild birds in this area, posing a potentail threat to both humans and animals.

Abstract 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 The macrolides appear to have considerable effects for treatment of bovine mastitis because of excellent diffusion into the mammary gland, long half-life, low protein binding, high intracellular concentration and lipid solubility. Acquired resistance to macrolides in Staphylococcus aureus is primarily related to target-site modification through acquisition of an erm gene. In the present study the prevalence of both phenotypic and genotypic tylosin resistance in S. aureus isolates (n = 103) from subclinical mastitis in nine dairy farms belonging to three different province of Iran were investigated. Overall, ermA, ermB and ermC was found in 7.80%, 32.00%, and 20.40% of S.aureus isolates, respectively. A very high percent of isolates (56.90%) were resistant to tylosin. MIC₉₀ and MIC₅₀ values were 64 and 32 µg mL^-1, respectively. Most of tylosin resistant isolates did not harbour any erm gene but ermB was dominant gene among 58 tylosin resistant isolates of S. aureus. In overall, tylosin resistance was prevalent in S. aureus isolates obtained from bovine mastitis in Iran.

Abstract Doogh is the most popular and commonly consumed yoghurt-based Iranian drink. The aim of this study was to investigate the antibacterial effects of Ziziphora clinopodioides essential oil (ZEO) at 0.10 and 0.20% concentrations, nisin at 250 and 500 IU mL^-1, and their combination against Salmonella typhimurium and Staphylococcusaureus in doogh during storage at 4 ˚C for 9 days. Nine batches were studied as follows: control: no ZEO or nisin added, A: 0.10% ZEO, B: 0.20% ZEO, C: 250 IU mL^-1 nisin, D: 500 IU mL^-1 nisin, E: 0.10% ZEO + 250 IU mL^-1 nisin, F: 0.10% ZEO + 500 IU mL^-1 nisin, G: 0.20% ZEO + 250 IU mL^-1 nisin and H: 0.20% ZEO + 500 IU mL^-1 nisin. Based on gas-chromatography and mass spectrometry, carvacrol (65.22%), thymol (19.51%), p-cymene (4.86%) and ɣ-terpinene (4.63%) were the major components of ZEO. The populations of S. typhimurium and S.aureus in samples treated with all concentrations of the ZEO and nisin were kept below 1 log CFU mL^-1 on day 5 of storage, while the count of S. typhimurium and S.aureus was found as 2.72 ± 0.02 and 2.21 ± 0.00 log CFU mL^-1 on day 5 for untreated samples, respectively. The ZEO separately and in combination with nisin, was very effective against these two common food-borne pathogens. The ZEO alone and in combination with nisin could be considered as a potential strong antimicrobial agent that can be used for the growth inhibition of aforementioned bacteria in food products especially doogh.