Prevalence of methicillin-resistant Staphylococcus aureus and pattern of antimicrobial resistance in mastitis milk of cattle in Chitwan, Nepal.

BACKGROUND: The threat of methicillin-resistant Staphylococcus aureus (MRSA) exists globally and has been listed as a priority pathogen by the World Health Organization. One of the sources of MRSA emergence is livestock and its products, often raised in poor husbandry conditions. There are limited studies in Nepal to understand the prevalence of MRSA in dairy animals and its antimicrobial resistance (AMR) profile. A cross-sectional study was conducted in Chitwan, one of the major milk-producing districts of Nepal, from February 2018 to September 2019 to estimate the prevalence of MRSA in milk samples and its AMR profile. The collected milk samples (n = 460) were screened using the California Mastitis Test (CMT) and positive samples were subjected to microbiological analysis to isolate and identify S. aureus. Polymerase Chain Reaction (PCR) was used to identify the presence of the mecA gene and screen for MRSA. RESULTS: In total, 41.5% (191/460) of milk samples were positive in the CMT test. Out of 191 CMT positive milk samples, the biochemical tests showed that the prevalence of S. aureus was 15.2% (29/191). Among the 29 S. aureus isolates, 6.9% (2/29) were identified as MRSA based on the detection of a mecA gene. This indicates that that 1.05% (2/191) of mastitis milk samples had MRSA. The antibiotic sensitivity test showed that 75.9% (22/29) and 48.3% (14/29) S. aureus isolates were found to be sensitive to Cefazolin and Tetracycline respectively (48.3%), whereas 100% of the isolates were resistant to Ampicillin. In total 96.6% (28/29) of S. aureus isolates were multidrug-resistant (MDR). CONCLUSIONS: This study revealed a high prevalence of S. aureus-mediated subclinical mastitis in dairy herds in Chitwan, Nepal, with a small proportion of it being MRSA carrying a mecA gene. This S. aureus, CoNS, and MRSA contaminated milk poses a public health risk due to the presence of a phenotype that is resistant to very commonly used antibiotics. It is suggested that dairy herds be screened for subclinical mastitis and treatments for the animals be based on antibiotic susceptibility tests to reduce the prevalence of AMR. Furthermore, future studies should focus on the Staphylococcus spp. to explore the antibiotic resistance genes in addition to the mecA gene to ensure public health.

Prevalence, antibiogram and risk factors of thermophilic Campylobacter spp. in dressed porcine carcass of Chitwan, Nepal.

BACKGROUND: Campylobacter is the primary cause of food borne gastroenteritis. Moreover, the emergence of multiple drug resistant campylobacters from poultry and pork has produced a potential threat to public health. Research addressing these issues is sparse in Nepal. So, this cross-sectional study aims at determining the prevalence, antibiogram and risk factors of campylobacters from dressed porcine carcass of Chitwan, Nepal. RESULTS: We collected 139 samples of dressed porcine carcass from 10 different pork shops located in Chitwan district and processed according to OIE Terrestrial Manual, 2008, chapter 2.8.10. Antibiogram of identified Campylobacter spp. was evaluated against nine commonly used antibiotics by using disc diffusion method following CLSI guidelines. The prevalence of Campylobacter spp. was 38.84% (C. coli 76% and C. jejuni 24%). There was no significant difference (p > 0.05) between the prevalence rate of male (32.4%) and female (41%) carcass. Ampicillin and erythromycin showed the highest resistance (92.59% each) followed by colistin (72.2%), tetracycline (61.1%), nalidixic acid and cotrimoxazole (44.4% each), ciprofloxacin (31.5%) and gentamicin (5.56%). Moreover, 77.8% of the isolates were resistant to more than two antimicrobials. Nalidixic acid and tetracycline showed significant difference (p < 0.05) in the resistivity pattern among different species of Campylobacters. The association between prevalence rate and regular sanitization of slaughter slab equipments was significant (p < 0.05). Similarly, prevalence rate was significantly associated (p < 0.01) with chilling and contamination of intestinal content with carcass. CONCLUSIONS: The pork meat of Chitwan is highly contaminated with antibiotic-resistant Campylobacters and slaughtering practices play significant role in contamination. It is necessary to train the butchers about hygienic slaughtering practice. The consumers as well as butchers should adopt safety measures to prevent themselves from antibiotic resistant campylobacters. The veterinary practitioners should adopt prudent use of antibiotics in pigs.

Effect of different synbiotic administration methods on growth, carcass characteristics, ileum histomorphometry, and blood biochemistry of Cobb-500 broilers.

Background and Aim: To combat enteric infections and antibiotic resistance in the poultry industry, researchers seek alternatives such as probiotics, prebiotics, and synbiotics as growth promoters. Synbiotics support probiotic growth through the supply of essential nutrients. The study's objectives were to assess the most effective delivery methods for synbiotics and evaluate their growth, histomorphometric, and hematological impacts on Cobb-500 broilers. Materials and Methods: Two studies, independently conducted, employed a completely randomized design. One hundred and eighty viable eggs in the first trial were assigned to three groups: Control (T1), sterile water (T2), and synbiotic in sterile water (T3). On the 21st day of hatching, hatchability, day-old body weights, and ileum samples for histomorphometric analysis were recorded. In the second trial, out of 500 viable eggs, 200 eggs were fed in ovo with synbiotics (PoultryStar® sol, Biomin Singapore Pte Ltd, Singapore) on 17.5 days and 300 were set aside without in ovo injection. The treatments were control (T1), in water synbiotic (T2), in ovo synbiotic (T3), combination of in ovo synbiotic and synbiotic in feed (T4), and synbiotic in feed only (T5). On 21 and 42 days, blood, ileum, and visceral organ samples were collected for laboratory analysis. Data on weight gain, daily feed intake, and water consumption were recorded for 42 days. Results: The initial experiment's results revealed a decrease in hatchability, slight weight increase, and significant intestinal morphological changes with the use of an in ovo synbiotic. Applying synbiotic through various methods in the second trial yielded better growth results, lower blood cholesterol, and significantly longer (p < 0.05) villi on 21 days. Conclusion: Using the in ovo method to administer synbiotics lowered hatchability. Use of synbiotics with any method or in combination enhances growth, ileum structure, dressing yield, feed efficiency, and cholesterol levels in blood. Synbiotics enhance gut health and overall performance in broilers when used through diverse approaches.

Virulence genes of avian pathogenic Escherichia coli isolated from commercial chicken in Nepal.

Colibacillosis is the most common bacteriological disease in poultry. The purpose of this study was to determine the recovery rate of avian pathogenic Escherichia coli (APEC) strains, the distribution, prevalence of Escherichia coli Reference (ECOR) collection and virulence associated gene (VAG) in four types of chickens infected by colibacillosis. Commercial broilers and layers had the highest percentage of positive APEC isolates (91%). We confirmed the ECOR phylogroup including B1 and E for the first time in Nepal. The prevalences of these phylogroups among chicken types were significantly different (p < 0.001). Among 57 VAGs, the number of genes found per isolate ranged from 8 to 26, with the top 5 VAGs being fimH (100%), issa (92.2%), traTa (90.6%), sit chro. (86%), and ironEC (84.8%). We found significant differences in gene prevalence among the chicken types. The predominance of B1 and E, and the VAG patterns suggest considering ECOR phylogroup and VAGs while formulating strategies for the prevention and control of APEC.

Reproduction of fowl typhoid by respiratory challenge with Salmonella Gallinarum.

Fowl typhoid is a disease of adult chickens and is caused by Salmonella Gallinarum infection via the alimentary tract. The experimental reproduction of fowl typhoid per os (PO) requires artificial conditions to minimize the effect of gastric acid, and several Salmonella serovars have been known to be transmitted via the respiratory route. Therefore, we have hypothesized the existence of a respiratory route for Salmonella Gallinarum infection and have attempted to reproduce fowl typhoid via intratracheal challenge. In accordance with our hypothesis, the intratracheal challenges of Salmonella Gallinarum reproduced exactly same lesions as fowl typhoid and induced higher mortality and morbidity than those of the PO challenge. Therefore, this study represents the first reproduction of fowl typhoid via respiratory route, and our findings may be useful for understanding the transmission of Salmonella Gallinarum in the field.