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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.

Live spore Sterne 34F-2 strain of Bacillus anthracis vaccine is being used to control anthrax disease in Bangladesh. After produced this vaccine in Livestock Research Institute (LRI) under the Department of Livestock Services (DLS), it is distributed at the farmers level through district and Upazila Livestock Offices. In these distribution pathways, the vaccine has been transported and stored for a few days in each station. The present study was carried out to evaluate the humoral immune response of the anthrax vaccine and to measure the impact of existing transportation and storage systems on immunity status. For that a total of 60 cattle were randomly selected , divided into three groups and used the vaccines collected from three distribution points. Serum samples were collected before and after the 1st month, 4th month, 7th 10th, and 13th month of vaccination respectively the anthrax antibody level in blood were monitored. The optical density was converted to ELISA units (EU) and used to express the antibody level in the vaccinated animals. It was significantly increased above the protection level (1.00) for a year. Before vaccination, the average ELISA unit of serum sample was 0.18± 0.01, after vaccination it was raised above the protective level (1.00) within one month and continued up to a year. In the chi-square test (95% confidence level), there was no significant difference (p

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: 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 fumigatusand Aspergillus nigerfailed 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. Adv. Vet. Anim. Res., 7(2): 338-344, June 2020

Natural colorants have been used in several ways throughout human history, such as in food, dyes, pharmaceuticals, cosmetics, and many other products. The study aimed to isolate the natural colorant-producing filamentous fungi Aspergillus niger from soil and extract pigments for its potential use specially for food production. Fourteen soil samples were collected from Madhupur National Park at Madhupur Upazila in the Mymensingh district, Bangladesh. The Aspergillus niger was isolated and identified from the soil samples by following conventional mycological methods (cultural and morphological characteristics), followed by confirmatory identification by a polymerase chain reaction (PCR) of conserved sequences of ITS1 ribosomal DNA using specific oligonucleotide primers. This was followed by genus- and species-specific primers targeting Aspergillus niger with an amplicon size of 521 and 310 bp, respectively. For pigment production, a mass culture of Aspergillus niger was conducted in Sabouraud dextrose broth in shaking conditions for seven days. The biomass was subjected to extraction of the pigments following an ethanol-based extraction method and concentrated using a rotary evaporator. Aspergillus niger could be isolated from three samples. The yield of extracted brown pigment from Aspergillus niger was 0.75% (w/v). Spectroscopic analysis of the pigments was carried out using a UV–VIS spectrophotometer. An in vivo experiment was conducted with mice to assess the toxicity of the pigments. From the colorimetric and sensory evaluations, pigment-supplemented products (cookies and lemon juice) were found to be more acceptable than the control products. This could be the first attempt to use Aspergillus niger extracted pigment from soil samples in food products in Bangladesh, but for successful food production, the food colorants must be approved by a responsible authority, e.g., the FDA or the BSTI. Moreover, fungal pigments could be used in the emerging fields of the food and textile industries in Bangladesh.

Objective: The present study was performed for isolation, identification, and molecular detection of Avipoxvirus [Turkeypox virus (TPV), Fowlpox virus (FPV), and Pigeonpox virus (PPV)] from field outbreaks in some selected areas of Mymensingh division, Bangladesh. Materials and Methods: A total of 60 suspected cutaneous nodular samples (10 TPV, 20 PPV, and 30 FPV) were collected. The samples were then subjected to isolation and identification by chicken embryo propagation followed by confirmation using polymerase chain reaction (PCR). Results: The TPV, FPV, and PPV were successfully isolated and identified from the nodular samples using embryo propagation and PCR technique targeting pox virus p4b gene. Out of 10 Turkeypox suspected field samples, five (50%) were positive for TPV. Similarly, among 30 Fowl pox suspected field samples, 12 (40%), and out of 20 Pigeonpox suspected field samples, eight (40%) were found to be positive for FPV and PPV, respectively. The overall prevalence of avipox (TPV, FPV, and PPV) virus infections in Mymensingh division was 41.67% (n = 25/60). Conclusion: This study has shown that TPV, FPV, and PPV are circulating in Mymensingh division. The isolated TPV, FPV, and PPV field isolates can be used as vaccine candidates to develop an effective vaccine for effective controlling of the avipox in Mymensingh division and surrounding areas. [J Adv Vet Anim Res 2019; 6(1.000): 54-59]

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 3.9×10 5 , 4.9×10 6 , 3.5×10 4 , 1.1×10 3 and 1.5×10 3 CFU/mL, while E. coli counts were 5.5×10 7 , 6.3×10 7 , 4.4×10 5 , 3.3×10 3 and 3.1×10 3 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

Among over 180 Aspergilli, Aspergillus fumigatus is the most common etiological agent causing invasive mold infection mostly in immunocompromised human and animal. Besides, the fungus is used for various useful purposes. However, for the utilization of A. fumigatus as a useful candidate, accurate identification is crucial. Here, the research work was aimed at identifying A. fumigatus from poultry feed samples using conventional and molecular techniques. Out of 23 feed samples, 2 (8.7%) were found to be positive for A. fumigatus. The internal transcribed spacer 1 (ITS 1) and ITS 2 regions and the 5.8S ribosomal DNA (rDNA) region of the fungus were amplified by polymerase chain reaction. The ITS regions are located between the 18S and 28S rRNA genes, and rRNA gene for 5.8S RNA separates these two ITS regions. The isolated gene has been sequenced and deposited in the GenBank (accession no. KC142152). The gene was 100% similar to other reference species of A. fumigatus, whereas in phylogenetic analysis, a clear distance was found in the cases of other Aspergilli. Based on the unique nature of the ITS1 and ITS2 regions and phylogenetic analysis of the genes, A. fumigatus was correctly identified. The isolated strain could be a good candidate for further studies especially for utilization in the field of biotechnology.Res. Agric. Livest. Fish.4(3): 165-171, December 2017

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. [J Adv Vet Anim Res 2016; 3(1.000): 45-50]

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. Adv. Vet. Anim. Res., 7(2): 360-366, June 2020

Vaccination is usually practiced to prevent and control anthrax in Bangladesh. For this purpose, vaccine prepared from Sterne F-24 strain ofBacillus anthracisby Livestock Research Institute (LRI), Mohakhali, Dhakahas long been used in this country. However, in some cases anthrax occurred in vaccinated animals in Bangladesh. A total of 100 cattle at LalTeer Livestock Research and Development Farm, LalTeerLivestock Limited, Bangladesh, aging between 3-6 years and weighing between 250-400 kg were randomly selected for vaccination purpose. Blood samples (n=100) were collected before the vaccination for collecting pre-vaccination serum, andthe animals were vaccinated (at 1 mL/animal; 1x107 spores/mL) with the anthrax vaccine produced by LRI. All blood samples from the vaccinated animals were collected on day 7, 28, 60, 90, 120, 150, 180, 240, 270, 300, 330, and 360 of post-vaccination, and serum samples were prepared. The antibody levels in the serum samples against anthrax were monitored using an Enzyme-Linked Immunosorbent Assay (ELISA). Over the course of 12 months, the antibody titers were found at the level higher than the reference value. Though there were reports on anthrax suspected cases in this farm, no such cases were reported during the study period. Thus, the vaccine appears to induce adequate antibody response against anthrax in Bangladesh.Microbes and Health, January 2015. 4(1): 36-38

Here, we report the first genome sequence of Pasteurella multocida BAUTB2 isolated from a buffalo that died from hemorrhagic septicemia in Rajshahi, Bangladesh. Using Illumina HiSeq technology, the BAUTB2 genome length was determined to be 2,439,149 bp, with 40.8% GC content, 2,307 coding sequences (CDS), 6 rRNAs, 51 tRNAs, and 4 noncoding RNAs (ncRNAs).

Pasteurella multocida type A is the etiologic agent of fowl cholera, a highly contagious and fatal disease of chickens. The present research work was performed for the isolation, identification and molecular detection of P. multocida Type A from chickens. Liver, heart and spleen of suspected dead chicken (n=35) were collected from Gazipur and Pabna districts in Bangladesh. The targeted bacteria from the samples were isolated, identified and characterized based on their morphology, staining, cultural, biochemical characters, pathogenicity test, histopathological study and Polymerase Chain Reaction (PCR). The P. multocida organism was isolated from 11.42% (n=4/35) samples. The organisms were gram negative, non-spore forming rod, non-motile, occurring singly or pairs in Gram staining, whereas in Leishman’s stain, bipolar shaped organisms were observed. All the isolates were found positive for oxidase and catalase tests, produced indole, and fermented glucose, mannitol and sucrose. Necrotic foci in liver and congestion with hemorrhages in heart were found on necropsy. After pathogenicity test, the pathological changes were reconfirmed by histopathology depicting congestion, hemorrhage and lymphocyte infiltration in heart, liver and spleen tissues. In type specific PCR reaction, the organisms were confirmed as P. multocida Type A. In conclusion, P. multocida type A is prevalent among poultry in the studied regions; thus, care must be taken to control of the disease. http://dx.doi.org/10.5455/javar.2015.b104

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.

Cytochrome P450 monooxygenases (P450s) are ubiquitous heme-thiolate proteins that have potential biotechnological application. Thermostable-P450s that can withstand hostile industrial conditions, such as high temperatures, extremes of pH and organic solvents, are needed for biotechnological usage. Here, for the first time, we report a large number of thermostable-P450s from two thermophilic biomass-degrading fungi, Myceliophthora thermophila and Thielavia terrestris. Genome-wide P450 analysis revealed the presence of 79 and 70 P450s (P450ome) in T. terrestris and M. thermophila. Authentic P450s containing both the P450 signature domains (EXXR and CXG) were classified as follows: T. terrestris (50 families and 56 subfamilies) and M. thermophila (49 families and 53 subfamilies). Bioinformatics analysis of P450omes suggested the presence of a large number of thermostable-P450s. Based on aliphatic index cut-off (>90), 14 and 11 P450s were determined to be thermostable in T. terrestris and M. thermophila. Among the thermostable P450s, six P450s from T. terrestris and three from M. thermophila had a melting temperature (Tm) of >65 °C, suggesting their hyperthermal tolerance. Analysis of the instability index of two ascomycete P450omes revealed the presence of 12 and 19 in vitro stable P450s in T. terrestris and M. thermophila. Overall, six P450s from T. terrestris and four from M. thermophila showed both thermal tolerance and in vitro stability. Thermophilic ascomycetes P450s are of potential interest from a structural, mechanistic and biotechnological point of view, as five P450s showed higher thermal tolerance and five showed higher in vitro stability compared to the well-characterized thermostable-P450s CYP175A1 (bacteria) and CYP119 (archaea).

Black quarter (BQ), caused by Clostridium chauvoei, is an economically important, highly infectious bacterial disease of cattle, goat and sheep. The present study was conducted to isolate and identify the causal agent of BQ from field cases. A total of 32 samples from necropsied areas of cattle presented to the Teaching Veterinary Hospital, Bangladesh Agricultural University were collected. The samples were subjected for bacteriological culture followed by identification through a series of conventional bacteriological techniques, staining properties and biochemical tests. The bacteria were cultured in anaerobic condition using candle jar. Morphological characters of bacteria were observed after Gram stain. The suspected samples were subjected for several passages in liver infusion broth to get pure culture. On neomycin blood agar, the bacteria showed hemolysis. Based on the cultural and staining characteristics and biochemical tests, 4 (12.5%) out of 32 samples were confirmed as Cl. chauvoei. It is concluded that Cl. chauvoei has been successfully isolated and identified from BQ affected cattle. [Fundam Appl Agric 2019; 4(1.000): 649-654]

Zika virus, a member of Flaviviridae is the etiology of Zika or Zika fever or Zika virus (ZIKV) disease characterized by mild symptoms similar to very mild form of Dengue or Chikungunya. The virus transmits through Aedes mosquitoes, particularly by Aedes aegypti. The most dangerous effect of ZIKV infection is the ability of the virus to cause microcephaly and congenital malformation to the newborn baby if the mother is infected. The neurological disorders including Guillain-Barre syndrome might be associated with adults and children due to ZIKV infections. Zika has emerged as a serious global public health problem as it has been found in 87 countries, particularly in Africa, America, and Asia and has no vaccine and treatment so far. Bangladesh is at a high risk of ZIKV infection and we consider ZIKV as a possible emerging threat for Bangladesh. This short review summarizes the insights of ZIKV infection, present status of the disease in Bangladesh and its neighboring countries, and recommendations for necessary preparations and strategies to be taken for effective controlling of the ZIKV infection in Bangladesh before getting any havoc. [J Adv Vet Anim Res 2019; 6(4.000): 575-582]

The present research work was carried out for the isolation and identification of bacterial flora from internal organs of broiler during the period from January 2012 to June 2012. Ten Hubbard classic broiler bird were purchased from retail market in Mymensingh, Bangladesh. The birds were sacrificed and their liver, lung, esophagus, duodenum and tracheal swab samples were collected (n=50). Using standard bacteriological techniques, Escherichia coli was isolated from 26 (52%) samples. Similarly, Salmonella spp. , Staphylococcus spp., Bacillus spp. , and Pasteurella spp. were isolated from 15 (30%), 10 (20%), 9 (18%) and 4 (8%) samples, respectively. On the basis of individual sample type, E. coli could be isolated from 8 (80%) duodenum samples being the most prevalent organism. On the other hand , Salmonella spp. , Staphylococci spp. , Bacillus spp. and Pasteurella spp. were identified in 5 (50%) lungs, 5 (50%) liver, 4 (40%) duodenum and 2 (20%) lungs samples, respectively. Among these isolated bacteria, E. coli was found to be pathogenic for mice. Antibiogram studies revealed that Ciprofloxacin was highly sensitive against all the isolated bacteria. Diversified bacterial species are prevalent in broiler. However, E. coli and Salmonella spp. infection might make the bird vulnerable for easy access of infection. Proper vaccination and use of selective antibiotics are crucial in protecting broilers from these pathogens. DOI: http://dx.doi.org/10.3329/mh.v1i2.14094 Microbes and Health, 2012 1(2): 72-75

A microbial consortium that reductively dechlorinates trichloroethene, cis-1,2-dichloroethene (cis-DCE), and vinyl chloride (VC) to ethene with methanogenesis was enriched from chloroethene-contaminated soil from Japan. Dechlorination activity was maintained for over 4 years. Using quantitative polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) analysis targeting the "Dehalococcoides" 16S rRNA gene, four strains were detected. Their growth and dechlorination activities were classified into two types: one that grows by converting cis-DCE to ethene and the other that grows by converting cis-DCE to VC. Then, the vcrA and bvcA genes encoding cis-DCE/VC reductive dehalogenases were detected. Inhibitors of methanogenesis (2-bromoethanesulfonate) and sulfidogenesis (molybdate) led to accumulation of cis-DCE and of VC respectively. These results suggest that methanogens and sulfate-reducing bacteria can play a significant role in dechlorination by "Dehalococcoides."

A functional library of cytochrome P450 monooxygenases from Aspergillus oryzae (AoCYPs) was constructed in which 121 isoforms were coexpressed with yeast NADPH-cytochrome P450 oxidoreductase in Saccharomyces cerevisiae . Using this functional library, novel catalytic functions of AoCYPs, such as catalytic potentials of CYP57B3 against genistein, were elucidated for the first time. Comprehensive functional screening promises rapid characterization of catalytic potentials and utility of AoCYPs.

Objective: The present research work was undertaken with the objectives to investigate the prevalence and molecular detection of the causal agents of sub-clinical mastitis (SCM) in cows at milk shed areas in Sirajganj and Pabna districts, Bangladesh. Materials and methods: A total of 300 milk samples were randomly collected from Baghabari milk shed areas of Sirajganj and Pabna districts. The milk samples were subjected for California Mastitis Test (CMT) for identifying SCM. Total 81 positive samples were then used for the isolation and identification of associated bacteria and fungi using conventional microbiological examination and biochemical tests, followed by confirmation by polymerase chain reaction (PCR) using specific primers. Besides, universal primers were used for amplification and sequencing of PCR products where specific primers were not used. Results: The overall prevalence of SCM was 51% (n=153/300). Based on bacteriological examination and biochemical tests, several bacteria were identified in this study; the orgnaisms included Staphylococcus sp. (45.68%) , Streptococcus uberis (14.81%) , Escherichia coli (9.88%), Proteus sp. (19.75%) , Salmonella sp. (1.23%) , Acinetobacter sp. (7.41%), and fungus (1.23%). PCR technique confirmed the bacteria as Staphylococcus aureus (279-bp), Streptococcus uberis (884-bp), E. coli (16SrRNA 585-bp, stx 1 606-bp, rfbO157 497-bp) and Salmonella sp. ( Inv-A gene796-bp). Conclusion: This study reveals that SCM in dairy cattle is persisting in Sirajganj and Pabna districts of Bangladesh. Hygienic practices should be improved, and providing technical intereventions may reduce the rate of SCM in the study areas. http://doi.org/10.5455/javar.2017.d238

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.

We explored the molecular diversity of cytochrome P450 genes in the filamentous fungus Aspergillus oryzae using bioinformatic and experimental approaches. Based on bioinformatic annotation, we found 155 putative genes of cytochromes P450 in the whole genome sequence; however, 13 of 155 appeared to be pseudogenes due to sequence deletions and/or inframe stop codon(s). There are 87 families of A. oryzae cytochromes P450 (AoCYPs), indicating considerable phylogenetic diversity. To characterize A. oryzae AoCYPs, we attempted to isolate cDNAs using RT-PCR and determined their transcriptional capabilities. To date, we have confirmed gene expression of 133 AoCYPs and cloned 121 AoCYPs as full-length cDNAs encoding a mature open reading frame. Using experimentally deduced sequences and intron-exon organization, we analyzed AoCYPs phylogenetically. We also identified intronic consensus sequences in AoCYPs genes. The experimentally validated exonic and intronic sequences will be a powerful advantage in identification and characterization of novel P450s from various ascomycetous fungi.

Duck plague (DP) is the most feared duck disease in the world. For isolation, identification, molecular detection and characterization of DP virus (DPV), a total of 94 samples were collected from commercial farms (n=6) and households (n=13) from Rajshahi (n=37), Netrokona (n=35) and Mymensingh (n=22) districts of Bangladesh. The samples were processed and inoculated into 11-13 days old embryonated duck eggs for virus propagation. Virus was identified using agar gel immunodiffusion test (AGIT) and passive hemagglutination (PHA) test, and was confirmed by polymerase chain reaction (PCR) targeting DNA polymerase and gC genes, followed by sequencing. Pathogenicity tests were performed using duck embryos, ducklings and ducks. Among the 94 samples, 17 isolates were confirmed as DPV by PCR amplification of partial DNA polymerase (446-bp) and gC genes (78-bp), respectively. One of the isolates (Anatid herpes 1 BAU DMH) was sequenced and found to be closely related with a Chinese variant of DPV (GenBank: JQ647509.1). Thus, we assume that both Bangladeshi and Chinese isolates of DPV may have a common ancestor. [J Adv Vet Anim Res 2015; 2(3.000): 296-303]