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Duck plague (DP) is one of the most important viral diseases which affects the duck population across the globe including Bangladesh. The present work was conducted to detect DP virus (DPV) from haor areas using a molecular-based approach, and compared with the contemporary isolate through molecular and phylogenetic analysis. For this purpose, 38 individual samples were collected from the Netrokona (n=20) district of the Mymensingh division and Kishoreganj (n=18) district of the Dhaka division. The identification of DVP was carried out by polymerase chain reaction (PCR) targeting DPV specific DNA polymerase genes followed by sequencing. PCR positive viral samples were used to propagate into 11-13 days old embryonated duck eggs through chorio-allantoic membrane (CAM) route for virus isolation. DPV were then propagated into duck embryo fibroblast (DEF) monolayer cell culture and confirmed by PCR. Among the 38 samples, 27 isolates were confirmed as DPV with the PCR amplicon size of 446 bp. Pathogenicity tests through the inoculation into day-old ducklings confirmed pathogenic strain. The PCR products of the isolated DPV specific DNA polymerase gene were sequenced commercially and submitted to GenBank (GenBank Accession No. KX768734.1). The sequence showed resemblance to isolates previously reported in India (GenBank Accession No. KX511893.1, KJ451479.1, KM012009.1), and China (GenBank Accession No. EF643559.1). Sequencing data also revealed nucleotide differences between Anatid herpes 1_BAU_DMH (previous report from our laboratory) and the present isolates. Further characterization, such as nucleotide and amino acid sequencing, would help to understand the strains along with its epidemiology. [ J Adv Biotechnol Exp Ther 2021; 4(1.000): 44-52]

Objectives: E. coli O157 is considered as one of the important pathogens causing subclinical mastitis in dairy cows. This study was undertaken to isolate E. coli O157 from the milk samples collected from subclinical mastitic cows using polymerase chain reaction (PCR) and investigate their antibiotic sensitivity patterns. Materials and Methods: 50 California Mastitis Test (CMT) positive milk samples were collected from apparently healthy crossbreed dairy cows in Baghabari, Sirajganj. For the enrichment and isolation of the organism, nutrient broth, MacConkey agar, and EMB agar were used. Later on, observing the biochemical tests result, all the isolates of E. coli were confirmed by PCR using genus-specific 16SrRNA primers. PCR-positive samples were then screened for the presence of the rfbO157 gene using gene-specific primers. The antibiotic sensitivity pattern of E. coli was assessed by the disk diffusion method against seven commonly used antibiotics. Results: Altogether, 8 (16%) isolates of E. coli were obtained, among which 5 (10%) were rfbO157 PCR positive. From the antibiotic sensitivity test, Gentamicin was the highest (75%) sensitive to the isolates, followed by Levofloxacin (62.5%), Cefixime (50%), Tetracycline (50%), and Ceftriaxone (25%). The highest resistance pattern was found against Ampicillin (100%) and Amoxycillin (87.5%). Conclusion: Raw milk containing E. coli O157 does not only reflect the status of the dairy herd. Additionally, it poses a serious threat to human health if it is consumed raw or used to make any type of value-added food product. [Vet. Res. Notes 2021; 1(2.000): 6-11]

Objectives: This study was designed to determine the Shiga-toxin producing genes and investigate antibiotic sensitivity patterns of the Escherichia (E.) coli isolated from ready-to-eat betel leaf from different roadside vendors. Materials and Methods: A total of 100 samples from the Bangladesh Agricultural University surroundings area were collected and screened for the detection of E. coli based on cultural, staining, and biochemical properties, followed by molecular characterization using genus-specific 16SrRNA primers by Polymerase Chain Reaction. stx-1 positive E. colis antimicrobial sensitivity pattern was determined by the disc diffusion method against ten antimicrobial agents. Results: According to this study, 44 out of 100 samples were confirmed to be associated with E. coli. Further, 44 positive E. coli were analyzed for stx-1 and stx-2 genes by duplex-PCR. From the results, two isolates were confirmed to be positive for the presence of the stx-1 gene, but no isolate was found to be positive for the stx-2 gene. The antimicrobial sensitivity test revealed that Ciprofloxacin was the most sensitive antimicrobial agent against the Shiga-toxin-producing E. coli (STEC), followed by Ceftriaxone and Levofloxacin. On the other hand, Amoxycillin and Cephradine were found to be highly resistant. Conclusion: Shiga-toxin-producing E. coli (STEC) is an important diarrheal pathogen that leads to watery to bloody diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome in humans. From the study, Levofloxacin and Ciprofloxacin can be recommended as effective drugs to control STEC infections in humans. [Vet. Res. Notes 2021; 1(2.000): 12-16]

Objectives: The goal of this study was to isolate, molecularly identify, and antibiogram of Staphylococcus aureus, as well as to investigate risk factors for subclinical mastitis in the Jhenaidah and Chuadanga districts of Bangladesh. Materials and Methods: A total of 100 milk samples were collected from apparently healthy goats at various farms in the Jhenaidah and Chuadanga districts. To detect samples with subclinical mastitis (SCM), the California Mastitis Test (CMT) was done. S. aureus was isolated and identified from milk samples that tested positive for CMT using a combination of standard bacteriological examinations, biochemical tests, and PCR confirmation. The disc diffusion test was used to determine the antibiogram status of the isolates. The risk factors for SCM in goats were analyzed using a semi-structured questionnaire with thirteen variables. Results: Some characteristics were significantly associated with goat mastitis, including farm, breed, shed construction, floor surface condition, and farm adviser. The CMT identified 52 positive samples for SCM, with 73.07% (n = 38/52) suggesting the presence of the nuc gene. Antibiotic susceptibility testing revealed that the isolated S. aureus was totally resistant to the antibiotics Penicillin, Oxacillin, and Vancomycin (21%). On the other hand, Tetracycline, Gentamycin, Norfloxacin, and Levofloxacin were all susceptible to all isolates. Conclusion: SCM is a severe problem in goats in Jhenaidah and Chuadanga districts. To minimize the risk of SCM infection in goats, hygiene precautions can be followed, and education among goat farm owners and farmers can be improved. [Vet. Res. Notes 2021; 1(2.000): 17-22]

Objectives: The purpose of this study was to isolate and identify Candida spp. and Saccharomyces spp. from bakery products (bread and cake) using a range of cultural (e.g., colony shape, size, color, elevation, surface edge, consistency, etc.) and morphological (e.g., cell shape, bud, etc.) features. Materials and Methods: Cake and bread samples (n = 40) were obtained from various retail out¬lets in Bangladesh Agricultural University (BAU), Mymensingh. The samples were collected asep¬tically and transferred to the Department of Microbiology and Hygiene at BAU, Mymensingh, for analysis. Processed samples were inoculated onto Potato Dextrose Agar media and incubated at a humidity of 75% for 5-7 days at 28°C. The fungi were primarily identified based on their colony structures. DNA was extracted from suspected colonies and was used for the detection of Candida spp. and Saccharomyces spp. using genus-specific primers (ITS-1 and ITS-4). Results: Candida spp. and Saccharomyces spp. had a combined prevalence of 50% (n = 20/40; p < 0.05). Candida spp. was identified in 15% (n = 6/40) of the fungi, while Saccharomyces spp. was detected in 35% (n = 14/40). Conclusion: In conclusion, it can be stated that bakery foods have a significant impact on human fungal infection in Bangladesh. [Vet. Res. Notes 2021; 1(1.000): 1-5]

Objective: The present study was carried out for the isolation, identification and antibiogram study of Salmonella spp. from apparently healthy Japanese quails (Coturnix japonica) at three different quail farms in Mymensingh, Bangladesh. Materials and methods: A total of 75 cloacal swab samples were randomly collected from apparently healthy Japanese quails from three different farms at Mymensingh, Bangladesh. The samples were subjected to a series of cultural and biochemical examination for the isolation of Salmonella followed by molecular detection by polymerase chain reaction (PCR). Motility of the Salmonella was performed by motility test and amplification of speF gene. The antibiogram profile of the isolates was also evaluated against commonly used antimicrobials by disc diffusion method. Results: The overall prevalence of Salmonella spp. in quails was found to be 13.33%(n=10/75). Out of the 10 isolates, seven were found to be motile. Farmwise, the prevalence of Salmonella spp. were 10%(n=3/30), 24%(n=6/25) and 5%(n=1/20) at the quail farms of Bangladesh Agricultural University (BAU), Shikarikanda and Akua, respectively. The antibiogram study revealed that all the isolates were resistant to both Erythromycin and Tetracycline. On the other hand, 100% isolates were sensitive to both Ciprofloxacin and Imipenem. Ninety percent isolates of Salmonella were resistant to Colistin sulphate. Neomycin was found to be sensitive to 80% Salmonella isolates. All the Salmonella isolates were found to be multidrug resistant (MDR). Conclusion: The presence of MDR Salmonella spp. in quails signifies public health importance of the organisms, which may be associated with food-borne illness. [J Adv Vet Anim Res 2018; 5(1.000): 60-66]

Salmonella is considered as a global problem ranking first among food borne diseases. All motile Salmonella of poultry origin are zoonotic and readily transmit to human via meat and eggs but reports on non - typhoidal Salmonella serovars circulating in layer chickens is very limited in South-East Asian countries including Bangladesh. Salmonella serovars isolated from apparently healthy chickens were characterized in the present study. Of 170 samples (cloacal swab 150 and feed 20) collected from commercial layer farms, motile Salmonella was isolated 4% (6/150) and 50% (10/20) respectively by cultural, biochemical, motility test and by detection of hisJ gene. About 5% (8/170) samples possessed serovar-specific gene fimA, suggesting that isolates were Salmonella enterica serovar Typhimurium. Antimicrobial susceptibility testing demonstrated that the isolated serovars were multidrug resistant. Therefore apparently healthy layer chickens harbour and transmit S. Typhimurium to the environment, although little is alarming since it has zoonotic significance and the isolates were resistant to commonly used first line of antibiotic in Salmonella infection.

Multidrug resistant (MDR) Salmonella are a leading cause of foodborne diseases and serious human health concerns worldwide. In this study we detected MDR Salmonella in broiler chicken along with the resistance genes and class 1 integron gene intl1. A total of 100 samples were collected from broiler farms comprising 50 cloacal swabs, 35 litter and 15 feed samples. Overall prevalence of Salmonella was 35% with the highest detected in cloacal swabs. Among the Salmonella, 30 isolates were confirmed as S. enterica serovar Typhimurium using molecular methods of PCR. Disk diffusion susceptibility test revealed that all the Salmonella were classified as MDR with the highest resistance to tetracycline (97.14%), chloramphenicol (94.28%), ampicillin (82.85%) and streptomycin (77.14%). The most prevalent resistance genotypes were tetA (97.14%), floR (94.28%), blaTEM-1 (82.85%) and aadA1 (77.14%). In addition, among the MDR Salmonella, 20% were positive for class 1 integron gene (intl1). As far as we know, this is the first study describing the molecular basis of antibiotic resistance in MDR Salmonella from broiler farms in Bangladesh. In addition to tetA, floR, blaTEM-1, aadA1 and intl1 were also detected in the isolated MDR Salmonella. The detection of MDR Salmonella in broiler chicken carrying intl1 is of serious public health concern because of their zoonotic nature and possibilities to enter into the food chain.

Anthrax, caused by Bacillus anthracis is an acute, febrile disease of warm blooded animals including humans. Social norms and poverty in addition to climatic factors such as soil conditions, seasons of year, ambient temperature and rainfall influence the persistence of the B. anthracis and anthrax outbreaks. The present study was designed to reveal the factors influencing the repeated outbreak of anthrax in Bangladesh. Considering the previous outbreaks of anthrax, Sirajganj, Bogra, Kushtia, Tangail and Mymensingh districts of Bangladesh were selected for this study. To elucidate the factors, qualitative data relating to the animal management, knowledge and behavior of the people; and quantitative data relating to soil conditions, ambient temperature and rainfall were acquired, and analyzed critically. Based on the outbreak histories, a year was divided into two seasons, anthrax prone season (May-November) and anthrax dry season (December-April). Anthrax spores could be isolated from 11.67% (n=14/120) of the soil samples collected from the study areas. The present study revealed that poor knowledge, lack of awareness, improper carcass disposal, inadequate vaccination, high Ca content and moisture in the soil along with high ambient temperature and rainfall during the anthrax prone season were the possible influencing factors of repeated outbreaks of anthrax in the study areas. Intensive propaganda to create public awareness of anthrax together with proper vaccination may reduce anthrax outbreaks in Bangladesh.

Objective: The methodology employed in this research was designed to identify and characterize the infectious bursal disease virus (IBDV) at the molecular level, originating from recent outbreaks in Bangladesh., Materials and Methods: The IBDV outbreak farm was investigated, and bursa of fabricius (BF) specimens were acquired from infected chickens. Initially, viruses in the processed samples were detected in chicken embryo fibroblast (CEF) cells, and the RT-PCR method was used to confirm IBDV. The positive samples were injected through chorioallantoic membrane route into the embryo of a 10-day-old specific pathogen-free (SPF) egg for virus isolation and pathogenicity testing. Finally, we sequenced the VP2 gene to identify phylogenetic relationships and detect mutations., Results: From the 77 collected samples, 42.85% (33/77) were found positive for cytopathic effects in CEF cells, and IBDV was detected in 31.16% (24/77) of the samples by RT-PCR. IBDV was isolated in SPF chicken embryos. In the pathogenicity test, infectious bursal disease was evident in seronegative chickens with visible signs of disease. Sequence analysis shows that the broiler-isolated viruses clustered with genotype A3B2 and backyard chickens with genotype A1B1. The presence of amino acid motifs for virulence markers was revealed in the partially sequenced VP2 gene with a mutation at S254G in four IBDV isolates from broilers. However, amino acids for virulence markers were absent in two isolates from backyard chickens, which shows sequence homology with IBDV classic strains., Conclusion: In this study, we identified and characterized circulating reassorted IBDV from vaccinated broilers, which may be one of the major causes of vaccination failure in broilers., Competing Interests: The researchers do not possess any competing priorities to disclose., (© The authors.)

Antimicrobial resistance (AMR) is a major health crisis globally. Migratory birds could be a potential source for antibiotic resistant (ABR) bacteria. Not much is known about their role in the transmission of ABR in Bangladesh. In this study, a total of 66 freshly dropped fecal materials of migratory birds were analyzed. Bacterial isolation and identification were based on cultural properties, biochemical tests, and polymerase chain reaction (PCR). The disk diffusion method was employed to evaluate antibiogram profiles. By PCR, out of 66 samples, the detection rate of Enterococcus spp. (60.61%; 95% confidence interval: 48.55-71.50%) was found significantly higher than Salmonella spp. (21.21%; 95% CI: 13.08-32.51%) and Vibrio spp. (39.40%; 95% CI: 28.50-51.45%). Enterococcus isolates were frequently found resistant (100-40%) to ampicillin, streptomycin, meropenem, erythromycin, and gentamicin; Salmonella isolates were frequently resistant (72-43%) to chloramphenicol, tetracycline, ampicillin, streptomycin, and erythromycin; and Vibrio spp. isolates were frequently resistant (77-31%) to vancomycin, ampicillin, erythromycin, tetracycline, and streptomycin. In addition, 60% (95% CI: 44.60-73.65%) Enterococcus spp., 85.71% (95% CI: 60.06-97.46%) Salmonella spp., and 76.92% (95% CI: 57.95-88.97%) Vibrio spp. isolates were multi-drug resistant (MDR) in nature. Three isolates (one from each bacterium) were found resistant against six classes of antibiotics. The bivariate analysis revealed strong associations (both positive and negative) between several antibiotic pairs which were resistant to isolated organisms. To the best of our knowledge, this is the first study in detecting MDR Enterococcus spp., Salmonella spp., and Vibrio spp. from migratory birds travelling to Bangladesh. Frequent detection of MDR bacteria from migratory birds travelling to Bangladesh suggests that these birds have the potential to carry and spread ABR bacteria and could implicate potential risks to public health. We recommend that these birds should be kept under an AMR surveillance program to minimize the potential risk of contamination of the environment with ABR as well as to reduce their hazardous impacts on health., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Author(s).)

Objective: This research work was conducted for the molecular characterization of the circulating foot-and-mouth disease (FMD) virus in Bangladesh and revealed out their serotype., Materials and Methods: The VP1 gene of six field isolates of FMD virus (FMDV) serotypes (two serotypes O, two serotypes A, and two serotypes Asia 1) was subjected for sequencing and phylogenetic analysis. Neighbor-joining trees were constructed by using the Molecular Evolutionary Genetics Analysis 6, having the field nucleotide sequences of FMDV and related sequences available in the GenBank., Results: The nucleotide sequences of the VP1 genes of serotypes O, A, and Asia-1 of the isolates revealed that overall isolates were 91%-100% similar to the isolates reported from Bangladesh and other neighboring countries. Among the isolates reported from Bangladesh, serotype O had 98%-100% identity, serotype A had 91%-100% identity, and serotype Asia-1 had 94%-100% identity. A phylogenetic analysis revealed that the FMDV serotype O PanAsia-02 sub-lineage was confirmed in Bangladesh under the Middle East-South Asian (ME-SA) topotype. On the other hand, we identified genotype VII (18) of Asia topotype (serotype A) and lineage C (serotype Asia-1)., Conclusion: The FMDV serotype O PanAsia-02 sub-lineage was confirmed in Bangladesh under the ME-SA topotype for the first time. The extensive cross-border animal movement from neighboring countries may act as the source of diversified FMDV serotypes in Bangladesh., Competing Interests: The author declares that they have no conflict of interests., (Copyright: © Journal of Advanced Veterinary and Animal Research.)

Objectives: Migratory birds play a major role in the transmission of pathogens globally, but still their role in the transmission of fungi in Bangladesh is not known. The present study was carried out for the isolation and molecular detection of fungi including Aspergillus from migratory birds traveling to Bangladesh., Materials and Methods: A total of 50 fecal samples were collected from BaojaniBaor, Magura, and areas close to Jahangirnagar University, Savar. The isolation of fungus was based on culture on Potato Dextrose Agar (PDA), followed by staining, morphology, and molecular detection using polymerase chain reaction (PCR)., Results: Among 50 samples, 40 showed positive for fungal growth on PDA, of which 30 yield only yeast-like colonies, five only molds, and five yielded both yeast and molds. The isolated molds produced various pigmented colonies, namely, black, whitish, grayish, olive green, and yellow. Among 10 molds, six were confirmed as fungi by PCR using genus-specific primers such as ITS1 and ITS4. Later, of these six fungi, five were confirmed as Aspergillus by PCR with primers such as ASAP1 and ASAP2 specific for Aspergillus genus. Therefore, the overall occurrence of Aspergillus was 10% (5/50). PCR specific for Aspergillus fumigatus and Aspergillus niger failed to produce specific PCR amplicon, suggesting that the isolated Aspergillus belongs to other groups., Conclusion: This is the first report describing the isolation and molecular detection of Aspergillus from fecal samples of migratory birds in Bangladesh. The present findings confirm that migratory birds are potential source for Aspergillus and other fungus in Bangladesh., Competing Interests: The authors declare that there is no conflict of interest in the publication of this article., (Copyright: © Journal of Advanced Veterinary and Animal Research.)