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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. [ABSTRACT FROM AUTHOR]

Objective: The research aimed to isolate, adapt to cell culture, and characterize the lumpy skin disease virus (LSDV) from clinically infected cattle in Bangladesh. Materials and Methods: From September 2019 to June 2020, 37 skin nodules and skin swabs were aseptically collected from afflicted cattle in the outbreak regions of Jhenaidah and Kishoreganj in Bangladesh. The LSDV was isolated from embryonated specific pathogen-free (SPF) chicken eggs along the chorioallantoic membrane (CAM) route and the Vero cell line after several blind passages. The viral attachment protein was targeted for molecular detection using polymerase chain reactions (PCR). For phylogenetic analysis, PCR-positive products were partially sequenced. Results: The virus was evident in the cell line, showed cytopathic effects after the 13 blind passage, and on the CAM of SPF chicken eggs, exhibited thickening of the CAM with pock-like lesions. A total of 12 samples (32.43%) tested positive for LSDV by PCR. Phylogenetic analysis of the present isolates (accession numbers MN792649 and MN792650) revealed 100% similarity with strains from India (MN295064), Kenya (AF325528, MN072619, KX683219), Greece (KY829023), Serbia (KY702007), and Kazakhstan (MN642592); moreover, 99.43% to 100% similarity to the sheep pox virus. Conclusion: Partially sequenced LSDV was developed as a vaccine seed and was first isolated in Bangladesh and characterized at the molecular level. [ABSTRACT FROM AUTHOR]

Objectives: This study was designed to isolate, identify, and determine the prevalence of Aspergilli in commercial chicken in selected areas of Bangladesh. Materials and Methods: A total of 50 lung samples from suspected dead chickens, comprising broilers (n = 32) and layers (n = 18), aged between 5 days and 45 weeks, were collected from poultry farms located in the Gazipur district in Bangladesh. Fungi were primarily identified based on the colony morphology using potato dextrose agar (PDA). DNA was extracted from the suspected colonies. Aspegillus spp. was detected by genus-specific ASAP-1 and ASAP-2. Aspergillus spp. were then screened by polymerase chain reaction targeting Aspergillus flavus (FLA-1 and FLA-2), Aspergillus fumigatus (ASPU and Af3r), and Aspergillus niger (ASPU and Nilr). Results: The overall prevalence of Aspergillus spp. was 44% (n = 22/50; p < 0.05). Among the Aspergilli, A. flavus was detected in 10% (n = 5/50) of the samples. Similarly, A. fumigatus and A. niger were detected at 26% (n = 13/50) and 8% (n = 4/50) respectively. Three samples were associated with more than one fungus; two fungi (A. flavus and A. niger) were in two samples, and three fungi (A. flavus, A. fumigatus, and A. niger) were in one sample. Conclusion: Isolation and prevalence of Aspergillus spp. in commercial chicken were studied for the first time in Bangladesh. [ABSTRACT FROM AUTHOR]

This research presents the application of glycine amino acid for establishing firm cell‐substrate interaction instead of expensive adhesion proteins, peptides and peptide derivatives. The glycine amino acid is chemically functionalized on the coverslip to achieve self‐assembled nanostructure. Glycine self‐assembly on NaCl treated coverslips is initiated with SiONa+:COO− linkage while their nanostructure is achieved with formation of glycine chain through NH3+:COO− covalent linkage between the adjacent molecules. The functionalization steps are confirmed by Fourier‐transform infrared spectroscopy (FTIR) investigation. The atomic force microscopy (AFM) and scanning electron microscopy (SEM) investigations reveal that glycine growth initiates at 4 Hours (H) post‐treatment while maximum growth appears after 8H‐10H. Both the vertical and horizontal growth of nanostructures show dependence on functionalization periods. Various levels of glycine functionalized surface show different levels of baby hamster kidney (BHK‐21) cell adhesion and proliferation efficiency with maximum performance for 10H functionalized surface. The adhesion and proliferation performance of 10H glycine functionalized surface shows negligible difference when compared with glycine‐aspartic acid (RGD) functionalized surface. Finally, growth curves obtained from both glycine and RGD functionalized surface reveal exponential growth phage up to 48H followed by stationary phage between 48H and 72H while death of many cells appears from 72H to 96H. Thus, this research concluded that glycine functionalized surface is equally effective for cell adhesion and proliferation. [ABSTRACT FROM AUTHOR]

Anthrax is one of the most important diseases that can be transmitted from cattle to humans, especially in countries with a high density of cattle. Cattle are usually infected by the ingestion of Bacillus anthracis spores with food ingredients. For that reason, the adequacy of anthrax spores in the soil is considered as one of the causes of anthrax in cattle. The present study is based on the identification of anthrax spores in the soil and the development of a map based on the presence of spores, so that the people of the community can take a practical step in this regard in selecting the place for their cattle to graze. For this purpose, soil samples were collected from the studied areas. After extraction of soil DNA with DNA extraction kits, all samples were tested using a qualitative detection method. For a rapid and reliable qualitative detection system for Pathogenic B. anthracis spores isolated from soil samples, the specific primer of two plasmids, pXO1 (gene Pag), pXO2 (genes Cap), and S-layer locus (gene Sap) were used. The presence of spores was found in 7 out of 50 samples. The results of a qualitative detection method of Bacillus anthracis spores in the pastureland inserted as per their GPS coordinates. Based on the presence of pathogenic anthrax spore in a specific location, a map was then prepared and displayed in the community. So that the cattle owners can decide where their cattle will graze and be careful to prevent spore contact of their animal. Among the various effective anthrax prevention measures, rich mapping according to the presence of spores can be helpful in community level that will protect people from anthrax disease in the future. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Objective: Shiga-toxin producing Escherichia coli (STEC) is the most important foodborne bacterial pathogen worldwide and the bovine animals are assumed as a reservoir of this pathogen. The present study was conducted to assess the role of bovine animals as the source of STEC. Materials and methods: To assess the role of bovine animals as the source of STEC, we examined 100 samples (50 rectal swab and 50 beef samples) collected from the local beef slaughterhouses by cultural, morphological, biochemical tests and polymerase chain reaction. Finally, the drug resistance pattern of isolated organisms has been examined. Result: In the preliminary screening by polymerase chain reaction (PCR), E. coli was more prevalent in rectal swab (n=21/50) than beef samples (n=16/50). Among 39 isolated E. coli, 10 isolates were confirmed as STEC (Rectal swab=7, Beef=3) by PCR, where stx2 gene (n=7/10) was predominant than stx1 gene (n=3/10). Remaining 29 isolates did not react to stx primers in PCR. Presence of STEC in beef samples was significantly associated with the fecal contamination at P≤0.1 (0.074818) in Pearson's correlation coefficient method. In addition, most of the isolated STEC strains were resistant to one or more commonly used antimicrobials in the country. Conclusion: The bovine animals and its products could be an important source of multidrug-resistant STEC in the country. [ABSTRACT FROM AUTHOR]

Objective: The present study was carried out to identify the associated Salmonella and Escherichia coli in betel leaves (Piper betle), and to develop an effective method to remove those microbes. Materials and methods: Betel leaves were collected from local and whole sale markets, and borouj (cultivation place). Salmonella and E. coli were isolated and identified by cultural, morphological, and biochemical tests followed by confirmation by polymerase chain reaction (PCR) targeting the genus specific 16S rRNA genes. Antibiogram of the isolated bacteria was performed by disc diffusion method. Different concentrations of Salmosan-A Soln were used to remediate the contaminating bacteria keeping the quality of betel leaves for longer periods. Results: Total Salmonella counts in the betel leaves were 3.9×105, 4.9×106, 3.5×104, 1.1×103 and 1.5×103 CFU/mL, while E. coli counts were 5.5×107, 6.3×107, 4.4×105, 3.3×103 and 3.1×103 CFU/mL in the betel leaves collected from K.R. market, Kewatkhali Bazaar, whole sale market, borouj in Kushtia and borouj in Natore, respectively. Antibiogram study revealed that the isolated bacteria were sensitive to doxycyclline, ciprofloxacin, chloramphenicol and cefotaxime. Application of 0.3% Salmosan-A Soln was found to be the most effective and suitable, where <1 CFU/mL in the betel leaves could be detected after treatment for 10 min. Conclusion: Results of this study indicated that treatment with 0.3% Salmosan-A Soln for 10 min is capable of removing Salmonella and E. coli from betel leaves keeping the treated leaves fresh up to 7 days of post-treatment. Method developed in this study can be adapted to reduce bacterial contamination of betel leaves more effectively in Bangladesh and other betel leaf exporting countries. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Objectives: The objectives of this study were to isolate and identify Pasteurella multocida from fowl cholera (FC) suspected chicken, and to prepare and efficacy determination of formalin killed fowl cholera vaccine using the isolated P. multocida strain. Materials and methods: A total of five suspected dead chickens were collected from Brothers Poultry Farm located at Gazipur district, Bangladesh. The samples were processed and the P. multocida was isolated through conventional bacteriological techniques, were finally confirmed by polymerase chain reaction using P. multocida specific primers targeting cap gene. The P. multocida isolate was used to develop a formalin killed fowl cholera vaccine. The efficacy of the newly prepared vaccine was determined in Starcross-579 chickens (n=30) aging 15 weeks either by injecting 1 mL (group-A; n=10) or 0.5 mL (group-B; n=10) vaccine containing approximately 3.2x108 CFU/mL P. multocida organism; 10 birds were kept as unvaccinated control. The sera from the vaccinated and control birds were collected and were subjected for antibody titre determination by enzyme-linked immunosorbent assay (ELISA). Finally the vaccinated birds were challenged using virulent strains of P. multocida to confer the protection against FC. Results: P. multocida could be isolated from both the samples. The formalin killed vaccine prepared from the isolated bacteria was subjected for the determination of antibody titre in chicken, and found that the antibody titres in the birds of group A and group B were 4.513 and 4.07 respectively after primary vaccination, and 4.893 and 4.37 respectively after booster vaccination. Most of the vaccinated birds were found to be survived after challenging with virulent strain of P. multocida. Conclusion: It is concluded that the causal agent of FC (P. multocida) was successfully isolated from FC affected dead chickens. The prepared formalin killed fowl cholera vaccine induces protective immune response and conferred protection against challenge infection caused by the virulent strain of P. multocida. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The study was conducted for the isolation and detection of Bacillus anthracis spores from soil collected from Sirajganj district (a north-western district of Bangladesh), and to assess the parameters that may relate to the repeated anthrax outbreak. A total of 48 soil samples were collected from the study area during January to November 2012. Endospores were extracted from soil and the Bacillus anthracis was identified using conventional bacteriological, biochemical and sensitivity test against Penicillin-G. The viable B. anthracis spores could be detected from 14 (29.17%) soil samples. Moisture content, pH, calcium and organic carbon contents of the soils were measured and the values of the endospore positive samples ranged from 6.31-28.37%, 5.17-7.22, 484.35-1372.35 ppm and 0.15-2.35%, respectively. All the endospore positive soil samples were of loamy type, while none of the clay type soil was found to be positive for B. anthracis, suggesting the influence of soil type on the occurrence of anthrax endospore in studied area. The mean pH of anthrax positive soil was weakly acidic (6.38±0.15), indicating that a suitable pH range for anthrax spore was present in the soil of Sirajganj. During the disease outbreak period (May and June) the average temperature of this area was 320C and the average rainfall was 158 mm and 90 mm, respectively. Although the temperature variation had no significant influence on the occurrence of anthrax spore, rainfall was found to be significant. [ABSTRACT FROM AUTHOR]