Epidemiology and molecular detection of Anaplasma spp. in goats from Chattogram district, Bangladesh.

OBJECTIVES: Anaplasmosis is an economically important disease affecting cattle, buffalo, sheep, goat etc. The study was conducted to determine the prevalence, potential risk factors and molecular identification of circulating Anaplasma spp. in goats in Chattogram district, Bangladesh. MATERIAL AND METHODS: Four hundred blood samples were collected from goats of different ages, breeds, sex, coat color and body condition. These goats were selected based on some inclusion criteria through the period of July 2017 to June 2018. Samples were examined microscopically (Giemsa staining method) followed by polymerase chain reaction (PCR) and sequencing to identify of Anaplasma spp. RESULTS: The overall prevalences were estimated 5.75% (23/400) and 15.75% (63/400) by microscopy and PCR, respectively. Anaplasma ovis (A. ovis) and Anaplasma marginale (A. marginale) were identified with the prevalence of 14.75% (59/400) and 1.0% (4/400), respectively through PCR. Among different risk factors, jamnapari breed (p = 0.027), no use of acaricide (p = 0.025) and presence of tick (p < 0.01) were found to be significantly associated with anaplasmosis. Sequence analysis of msp4 gene revealed that, Anaplasma spp. detected in the present study were highly similar with those of China, Venezuela, Mongolia, Spain, Tunisia, Cyprus, Italy, Brazil, Argentina, Australia, Japan and Columbia. CONCLUSIONS: In conclusion, strategic use of acaricide can control tick that ultimately will control the anaplasmosis in goats. Besides, rearing local goats in compare to cross and exotic breed are also recommended for the farmer to prevent the disease.

Study on bacterial pathogens through multiplex polymerase chain reaction system and their antimicrobial resistance pattern in goats presumed with fever and/or diarrhea.

BACKGROUND AND AIM: Goat is one of the major livestock species, plays an important role in the economy of Bangladesh. However, the outbreak of different infectious diseases in goats causes high mortality and economic losses due to lack of proper diagnosis and treatment. Conventional culture-based methods for detecting specific pathogens as confirmatory diagnosis are laborious as well as time-consuming in comparison to multiplex polymerase chain reaction (mPCR), by which multiple pathogens can be detected at a time. The present study was aimed to perform faster molecular identification of bacterial pathogens from goats presumed with fever and/or diarrhea and their antimicrobial resistance (AMR) pattern. MATERIALS AND METHODS: A total of 200 blood samples were collected from goats at S. A. Quaderi Teaching Veterinary Hospital (SAQTVH) in Chattogram Veterinary and Animal Sciences University for the period of July 2017-April 2018. DNA was extracted and subsequently, mPCR assay was performed for the screening of several bacterial pathogens (Salmonella spp., Listeria monocytogenes, Bacillus cereus, Yersinia enterocolitica, Campylobacter jejuni, Campylobacter coli, Clostridium perfringens, Vibrio cholerae, and Staphylococcus aureus). An antimicrobial susceptibility test against ten antimicrobials for positive samples of each organism was conducted using the Kirby-Bauer Disk-Diffusion Method on selective media. RESULTS: S. aureus, C. perfringens, L. monocytogenes, and Salmonella spp. were detected from collected samples and their overall prevalence was 11.5%, 3.5%, 1%, and 20.5%, respectively. The most common clinical signs were mild fever, nasal discharge, dyspnea, and coughing (39.1%) for S. aureus, diarrhea, convulsion, abdominal pain, and incoordination (57.1%) for C. perfringens, fever, protrusion of tongue, and incoordination (100%) for L. monocytogenes, and fever, anorexia, dehydration with mucous feces (36.6%) for Salmonella spp. infection in goats. AntimGentamicinicrobial diagram of S. aureus showed resistance against Cefotaxime (74%), Cefixime (65%), and Tetracycline (65%); highly sensitive against Amoxicillin (48%), Ciprofloxacin (44%), and Gentamicin (44%). On the other hand, C. perfringens showed highly resistant against Ampicillin (71%), Gentamicin (71%), sensitive against Penicillin (57%), and Cefotaxime (57%). L. monocytogenes were found to be sensitive to Penicillin (100%) and Cefixime (100%) and Salmonella spp. showed resistance to Ampicillin (78%) and Amoxicillin (59%) but sensitive to Ciprofloxacin (54%). CONCLUSION: This study identified pathogens with their specific clinical signs in goats presumed fever and/or diarrhea through mPCR with their AMR pattern in SAQTVH, Chattogram. Potential risk factors, measuring the strength of association of disease caused by these particular pathogens, were also determined. mPCR may use as an effective tool for rapid detection of pathogens in animal.

Molecular Epidemiology and Characterization of Theileria in Goats.

The study was carried out to determine the prevalence and associated risk factors of theileriosis in goats of Chattogram district, Bangladesh. Molecular characterization of circulating Theileria in this area was also undertaken. A total of 400 samples were collected from goats of different breeds, ages and sex with relevant information of rearing and management. The prevalence of theileriosis was 8.50% (34/400) by polymerase chain reaction though all of those samples were test-negative by microscopic examination. Among different risk factors season, breed and tick infestation were found to be significantly (p ≤ 0.05) associated with the prevalence of theileriosis in goats. Serous nasal discharge and swollen lymph nodes were determined to be significant clinical signs. The Theileria spp. detected in the present study closely resemble isolates which were previously detected in Myanmar and China. Further large scale epidemiological studies are required to identify the circulating species and responsible vectors, which would facilitate control measures for this disease in Bangladesh.

Estimation of the prevalence and determination of risk factors associated with demodicosis in dogs.

OBJECTIVES: Demodicosis is a vital skin problem in dogs. The present study has determined the prevalence and associated risk factors of demodicosis in dogs and the response to treatment. MATERIALS AND METHODS: A total of 100 skin scrapings were collected from dogs having dermatological lesions brought to the Teaching and Training Pet Hospital and Research Center of Chattogram Veterinary and Animal Sciences University for treatment purpose. The collected scrapings were dissolved in 10% potassium hydroxide to detect mites through microscopic examination. Various risk factors like breed, age, sex, hair type, health status, and management system (indoor and outdoor) were analyzed using the logistic regression model. Positive cases were treated with oral ivermectin (Scabo®; at 0.6 mg/kg/day) along with amitraz 12.5% (Ridd®) diluted to 0.05% for rubbing on the body after bathing with chlorhexidine shampoo (PetHex®). Clavulanate amoxicillin (Moxaclav®) and omega-3 fatty acids (OMG-3®) were also suggested to prevent secondary bacterial infection and to maintain skin and hair coat integrity. The recovery rate was observed every 2-4 weeks of treatment upto 11-13 weeks. RESULTS: The overall prevalence of demodicosis was recorded as 27%. Hair type, health status, and management system were significantly (p < 0.05) associated with the occurrence of demodicosis in dogs. Following treatment, the first negative skin scraping for mite was noticed at 8-10 weeks of treatment, and in all cases, clinical signs completely disappeared at 80-90 days of treatment. CONCLUSION: A good percentage of the dogs having dermatological lesions was determined as demodicosis. Long-term oral ivermectin and topical amitraz, an oral antibiotic, and nutritional therapy are effective against canine demodicosis. Keeping long-haired dogs with good body condition and indoor management is highly suggestive of preventing and controlling the disease.

Multidrug Antimicrobial Resistance and Molecular Detection of mcr-1 Gene in Salmonella Species Isolated from Chicken.

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.