Veterinary Research Forum

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 The aim of this study was to investigate the probiotic potential of autochthonous Lactobacillus species isolated from buffalo calves against multidrug-resistant Escherichia coli. A total of 252 rectal swabs were collected from healthy neonatal buffalo calves under 30 days old from six districts of Andhra Pradesh, India in a completely randomized design from August 2019 to August 2021, of which 190 Lactobacillus strains were isolated based on cultural, morphological, biochemical and molecular tests. Out of 190 isolates, 57 showed high levels of auto-aggregation (> 80.00%) and hydrophobicity (> 60.00%) and 51 of the 57 isolates had a zone of inhibition greater than 15.00 mm in diameter against multidrug-resistant E. coli in an Agar well diffusion assay. Among the 51 isolates, 36 were found to be acid and bile tolerant and showed varying levels of sensitivity to antibiotics such as erythromycin, clindamycin, tetracycline, chloramphenicol, and ampicillin. Among the 36 isolates, Limosilactobacillus reuteri 178, L. reuteri 209, L. fermentum 182, L. fermentum 211, and Lactiplanti-bacillus plantarum 34 were non-hemolytic, and none of the isolates were able to hydrolyse gelatine. Therefore, these five autochthonous Lactobacillus species may be used in probiotic or synbiotic formulations against multidrug resistant E. coli in buffalo calves.

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 this study was to compare the sedative and cardiovascular effects of the combination of xylazine-acepromazine versus xylazine-pregabalin - in horses. Four healthy crossbred horses were included in the study and assigned to two treatments. In treatment I (T1), the animals received xylazine hydrochloride (1.00 mg kg^-1) in combination with acepromazine maleate (0.05 mg kg^-1) intravenously. In treatment II (T2), the animals received intragastric administration of pregabalin (4.00 mg kg^-1) followed by xylazine hydrochloride (1.00 mg kg^-1) intravenously after 60 min. Head height above ground (HHAG) and echocardiographic indices were evaluated. In T1, recordings were made 5 minubefore and 5, 15, 30, 60, and 90 minu after drug administration. In T2, recordings were made 5 min before pregabalin, 55 minu after pregabalin administration, and then 5, 15, 30, 60, and 90 min after xylazine hydrochloride acepromazine injection. Analyses of the data showed there were no significant differences regarding HHAG and echocardiographic indices between the two treatments. Intragastric administration of pregabalin prior to xylazine could be considered as an alternative premedication regimen when acepromazine administration is contraindicated or undesirable.

Abstract The ostrich (Struthio camelus) is an important wild species highlighted in national and international livestock industry. This research was conducted to analyze the development of the ostrich respiratory system during fetal and embryonic stages. A total of 50 fertile ostrich eggs were collected from commercial farms and then incubated at 36.00 - 37.00 ˚C and 25.00 ± 2.00% humidity for 40 days. Sections were taken on days 13, 22, 26, 30, 36, and 42 of incubation from the lung and the cranial, middle, and caudal parts of the neck after decapitation of ostrich embryos and blood drainage. After fixation, processing, blocking, and sectioning, all samples were stained by Hematoxylin and Eosin, Alcian Blue (AB), Van Gieson, and Periodic acid-Schiff (PAS) techniques. It was concluded that the trachea in the 13-day-old embryo and goblet cells (PAS-positive and AB-positive) had incomplete rings of hyaline cartilage and differentiation of mesenchymal to the loose connective tissue. The bronchial stage of the lung was observed in the 22-day-old embryo, pseudoglandular stage in the 26-day-old embryo, and parabrachial and air capillary stage in the 30-day-old embryo. The information obtained from this study will be useful for diagnosing pathologies affecting this vital system and results in improving industrial breeding management.

Abstract In November 2021, an investigation was conducted into an outbreak of abortion, stillbirth, and the birth of calves with congenital abnormalities (arthrogryposis and hydranencephaly) at a dairy farm in Dasht-e-Mughan city, Ardabil province. A total of 70 cows experienced these issues. To determine the cause of the outbreak, post-mortem brain tissue samples were collected from two calves affected by hydranencephaly, which occurred shortly after their birth. Polymerase chain reaction (PCR) testing was conducted for multiple viruses, including bovine viral diarrhea (BVD), border disease, Akabane, Schmallenberg, and bluetongue viruses (BTVs). The samples were positive only for Akabane virus. Serum samples were collected from a group of 60 cattle, consisting of 45 adult cows and 15 younger calves aged between 8 to 10 months. These samples were analyzed to detect the presence of antibodies against the Akabane and Schmallenberg viruses. Both of these viruses are known to be responsible for causing abortion, stillbirth, and congenital abnormalities in calves. Among 45 cows that tested by competitive enzyme-linked immunosorbent assay (cELISA), 26.66% and 33.33% exhibited antibodies against Akabane and Schmallenberg viruses, respectively. Notably, 20.00% of cows co-exhibited antibodies for both viruses. Despite PCR evidence implicating Akabane virus as the principal etiology of clinical signs observed in the affected herd, the high co-seropositivity to Schmallenberg virus, warrants a thorough investigation into potential viral interactions. Further research is required to determine the source of the virus and their transmission routes. This information could facilitate the refinement of disease control strategies and improving the management of reproductive challenges in such affected herds.

Abstract The poultry products are known as a source of zoonotic and multi-drug resistant pathogens, especially Salmonella spp. The objective of this study was using bacteriophages as an alternative anti-microbial agent against Salmonella typhimurium isolate from turkey poults. The antibiotic susceptibility test was used to identify the antibiotic resistance pattern of the isolates. The bacteriophage was purified, enhanced and titrated using the Spot test and double layer agar (DLA) techniques after being isolated from a chicken slaughterhouse and sewage treatment facility. By determining the morphological characteristics of resulting plaque, the specificity and host range of the phage were studied on S. typhimurium isolates. A total number of 22 suspected Salmonella isolates were confirmed biochemically positive in sample by cultures method. Nine of these isolates (40.90%) were identified as S. typhimurium by polymerase chain reaction. All of isolates (100%) were resistant to chloramphenicol, doxycycline, kanamycin, florfenicol, rifampin, and erythromycin. Seven isolates (77.77%) were resistant to amoxicillin and nalidixic acid. The plaques were present with 3.00 ± 0.22 mm in diameter on the culture of 6 out of 9 (66.66%) isolates of S. typhimurium on brain heart infusion broth using DLA method. The amount of phage titer was 7.60 × 10⁷ phage forming unit mL^-1 and its multiplicity of infection value was calculated as 5.06 × 10^-2 based on obtained results. In place of antibiotics, the multi-drug resistant (MDR) S. typhimurium was successfully destroyed by the isolated bacteriophage from wastewater. In vitro settings were used in this investigation to identify the efficient bacteriophages against MDR S. typhimurium.

Abstract Since decades, Newcastle disease (ND) has become endemic in the poultry population of the Indian subcontinent. ND is a highly contagious disease of poultry and other avian species. However, the genetic nature of ND viruses circulating in the rock pigeons is unraveled. The present investigation is a part of Newcastle disease virus (NDV) surveillance in wild birds. Two velogenic NDV strains could be isolated from apparently healthy rock pigeons, thus establishing the status of carrier/reservoir host. The fusion protein cleavage site in the fusion protein has multiple basic amino acid (RRRKRF) motifs similar to velogenic isolates. Phylogenetic analysis based on complete fusion gene sequences confirmed that the isolates belong to NDV sub genotype XIII 2.2. Further analysis revealed several amino acid substitutions in the hypervariable region, heptad repeat regions and neutralizing epitopes of the fusion protein and heptad repeat regions and antigenic sites of the hemagglutinin-neuraminidase (HN) protein that are critical for fusion. A unique D170A substitution in the neutralizing epitope is identified that is critical for structure and function of the fusion protein. Mutations within the virulence determinants including fusion (F) and HN, elucidate continuous evolution of the viruses among the rock pigeons. Accidental spillover of these mutated viruses into commercial poultry operations may result in disease outbreaks with economic breakdown.