Subclinical mastitis (SCM), a silent threat in the dairy sector of Bangladesh poses a significant economic impact and serves as a potential source of infection for healthy cows, hindering efforts to achieve milk self-sufficiency. Despite the importance of this issue, limited research has been conducted on mastitis in Sylhet region of Bangladesh. This study aimed to investigate the molecular prevalence, antimicrobial susceptibility profile and resistant genes detection on pathogens (Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae) causing SCM. In a cross-sectional study utilizing convenience sampling, 325 milk samples were collected from apparently healthy dairy cows. Initially, SCM was detected using the modified Whiteside test (MWST) method. Suspected positive samples were then subjected to bacteriological culture and standard biochemical assays, followed by molecular identification through polymerase chain reaction (PCR). Finally, antimicrobial susceptibility testing was conducted on all PCR-positive samples using the Kirby-Bauer disk diffusion method on Mueller-Hinton agar. In the Sylhet district, the prevalence of SCM was 64.92 % (211 out of 325) at the individual animal level and 82.69 % (43 out of 52) at the herd level. Among the SCM-positive animals, S . aureus was found in 31.75 % (67 out of 211) of cases, E . coli in 81.99 % (173 out of 211), and K . pneumoniae in 66.82 % (141 out of 211). K . pneumoniae had the highest prevalence at 60 % in Zakiganj, S . aureus at 45 % in Zakiganj, and E . coli at 72 % in Bishwanath Upazila. Extended spectrum beta lactamase (ESBL) genes bla_TEM, bla_OXA, bla_CTX-M1,bla_CTX-M2,MultiCase_DHA were identified. Additionally, antibiotic resistance genes tet(A), AAC (3)-iv, and Sul1 were also detected. The pathogens exhibited resistance to Penicillins (ampicillin, amoxicillin), Cephems (cefuroxime, ceftazidime), and Tetracyclines (tetracycline). However, all three bacteria were highly sensitive to meropenem, amikacin, gentamicin, ciprofloxacin, and sulfamethoxazole-trimethoprim. These findings highlight the critical need for targeted antimicrobial stewardship and effective control measures to mitigate the impact of SCM on dairy production and animal health in the Sylhet region of Bangladesh.

The study was conducted to evaluate the prebiotic effects of Spirulina as a growth and immunity promoter for broiler chickens. Birds (n=100) were randomly and equally distributed into four groups (T0, T1, T2 and T3) and fed on a diet containing 0, 2, 4 and 8 g Spirulina/kg feed respectively for 4 weeks. The body weight was significantly (P<0.05) increased in the treatment groups fed with Spirulina diet from 7th days to 28th days old. FCR was also significantly (P<0.05) decreased among the treatment groups. Hematological parameters were significantly (P<0.05) increased except ESR which was decreased significantly (P<0.05) in the treatment group. Aspartate aminotransferase (ALT) and alanine aminotransferase (AST) level were significantly (P<0.05) decreased in all the treatment groups. The study suggests that, Spirulina is a good natural feed additive which has a tremendous effect to improve the broiler production and thereby may reduce the production cost. [J Adv Vet Anim Res 2015; 2(3.000): 304-309

In the present study, a total of 720 samples were collected from retail raw meat from 13 upazilas in Sylhet District, Bangladesh, of which 225 samples were from cattle meat, 210 samples were from goat meat, and 285 samples were from chicken meat. Salmonella enterica serovars Typhimurium and Enteritidis were screened for extended-spectrum β-lactamase (ESBL) genes using multiplex PCR. Among the 720 samples, Salmonella spp. was detected in 28.06% (202 out of 720) of the samples, with S. Enteritidis and S. Typhimurium were identified in 11.53% (83 out of 720) and 12.22% (88 out of 720) of the samples, respectively. It was found that all Salmonella enterica serovars isolated from cattle meat displayed multidrug resistance (MDR) based on antimicrobial susceptibility testing. Notably, a significant proportion of S. Enteritidis isolates and all S. Typhimurium isolates from goat meat demonstrated complete resistance to multiple drugs (ampicillin, cefuroxime, and ceftazidime). Regarding chicken meat, out of 89 isolates encompassing both S. Typhimurium and S. Enteritidis, 57 isolates (64.04%) exhibited MDR. Additionally, bla_CTX-M-1 exhibited the highest occurrence at 15.69% for S. Typhimurium and 7.89% for S. Enteritidis in chicken meat. Moreover, bla_CTX-M-9 was only detected at 3.92% for S. Enteritidis in chicken meat. Furthermore, bla_OXA had the highest prevalence rate of 19.04% for S. Enteritidis and 25.80% for S. Typhimurium in cattle meat, followed by chicken meat. These findings highlight the urgency for monitoring ESBL-producing Salmonella in retail raw meat and the need for strict measure to manage antibiotic use to prevent the spread of multidrug-resistant and ESBL-producing Salmonella strains, thereby protecting humans and reducing public health risks.

This study aimed to characterize virulence and antibiotic resistance genes in multidrug-resistant (MDR) and extensively drug-resistant (XDR) Escherichia coli and Klebsiella pneumoniae isolated from cases of subclinical mastitis (SCM) in buffaloes. A cross-sectional study was conducted on 1540 quarter milk samples collected from 385 buffaloes. Milk samples were screened using the California Mastitis Test and Modified Whiteside Test. Positive samples underwent bacterial culture, biochemical tests, biofilm detection and molecular analysis for pathogen identification and detection of virulence, resistance, and extended-spectrum beta-lactamase (ESBL) genes. The prevalence of SCM was 67.9% (1046/1540) at the quarter level and 80.8% (311/385) at the animal level. E. coli was identified in 9.5% (146/1540) of the samples, while K. pneumoniae was detected in 9.09% (140/1540). Virulence genes, such as stx1 (27.4%), and resistance genes, including aac(3)-iv (77.4%) and tetA (76.7%), exhibited higher prevalence. Additionally, β-lactamase genes, notably bla_TEM (67.1%), and ESBL genes, such as bla_CTX-M1, were detected. Biofilm formation was detected in 83.6% (122/146) of E. coli isolates and 75.7% (106/140) of K. pneumoniae isolates. Antimicrobial susceptibility testing revealed significant resistance to ampicillin, amoxicillin-clavulanic acid, and aminoglycosides. MDR was observed in 31.5% of E. coli and 39.3% of K. pneumoniae isolates, with XDR rates of 8.9% and 12.9%, respectively. These findings underscore the alarming spread of resistant pathogens in SCM-affected buffaloes, emphasizing the urgent need for ongoing surveillance and targeted intervention strategies.

Colistin (polymyxin E) is widely used in animal and human medicine and is increasingly used as one of the last-resort antibiotics against Gram-negative bacilli. Due to the increased use of colistin in treating infections caused by multidrug-resistant Gram-negative bacteria, resistance to this antibiotic ought to be monitored. The study was undertaken to elucidate the molecular mechanisms, genetic relationships and phenotype correlations of colistin-resistant isolates. Here, we report the detection of the mcr-1 gene in chicken-associated Salmonella isolates in Bangladesh and its in-silico functional analysis. Out of 100 samples, 82 Salmonella spp. were isolated from chicken specimens (liver, intestine). Phenotypic disc diffusion and minimum inhibitory concentration (MIC) assay using different antimicrobial agents were performed. Salmonella isolates were characterized using PCR methods targeting genus-specific invA and mcr-1 genes with validation for the functional analysis. The majority of the tested Salmonella isolates were found resistant to colistin (92.68%), ciprofloxacin (73.17%), tigecycline (62.20%) and trimethoprim/sulfamethoxazole (60.98%). When screened using PCR, five out of ten Salmonella isolates were found to carry the mcr-1 gene. One isolate was confirmed for Salmonella enterica subsp. enterica serovar Enteritidis, and other four isolates were confirmed for Salmonella enterica subsp. enterica serovar Typhimurium. Sequencing and phylogenetic analysis revealed a divergent evolutionary relationship between the catalytic domain of Neisseria meningitidis lipooligosaccharide phosphoethanolamine transferase A (LptA) and MCR proteins, rendering them resistant to colistin. Three-dimensional homology structural analysis of MCR-1 proteins and molecular docking interactions suggested that MCR-1 and LptA share a similar substrate binding cavity, which could be validated for the functional analysis. The comprehensive molecular and in-silico analyses of the colistin resistance mcr-1 gene of Salmonella spp. of chicken origin in the present study highlight the importance of continued monitoring and surveillance for antimicrobial resistance among pathogens in food chain animals.

Background: The most prevalent probiotic bacterium employed in the food industry is Lactobacillus because it can produce metabolites with antibacterial capabilities and exhibits hostility towards infections and microorganisms that cause spoilage.Aim: This study set out to identify naturally occurring Lactobacillus and plantaricin (pln EF) coding genes in raw cow milk and to assess the antibacterial potency of isolated Lactobacillus isolates.Methods: Following enrichment in De Man, Rogosa and Sharpe (MRS) broth, single colonies were isolated, and pure colonies were obtained by streaking on MRS agar. The 16S rRNA gene was amplified using polymerase chain reaction (PCR) to confirm the cultural positivity of all isolates. Additionally, the presence of plantaricin was verified by targeting the pln EF gene through PCR.Outcome: Out of the 166 raw milk specimens acquired from cows, 153 (91.17%; CI: 86.98–95.76) were identified as positive for Lactobacillus through both culture and biochemical screening. Subsequently, 121 (72.89%; CI: 65.46–79.49) of the isolates were affirmed to harbour Lactobacillus through PCR analysis. Within this subset, 6 isolates (4.96%; CI: 1.84–10.48) were found to possess the plnEF gene. When exposed to Lactobacillus isolates, Salmonella Typhimurium and Salmonella enterica displayed an average maximum zone of inhibition with a diameter measuring 24 mm. In contrast, Escherichia coli exhibited an average minimum zone of inhibition, featuring a diameter of 11 mm. Additionally, the Lactobacillus isolates demonstrated inhibitory zones against Staphylococcus aureus, Klebsiella pneumoniae and Klebsiella oxytoca, measuring 14, 22 and 19 mm, respectively.Clinical significance: Lactic acid bacteria, particularly Lactobacilli, are plentiful in cow milk and possess broad‐spectrum antibacterial properties.