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Abstract Trained immunity is a phenomenon in which brief exposure to an infectious agent or a vaccine can induce long-lasting changes in the host's immune system, enhancing protection against subsequent infections. The concept of trained immunity has a significant impact on the field of immunology and has the potential to revolutionize how we approach vaccination and infectious disease control. Investigations into trained immunity are rapidly advancing and have led to the development of new vaccines and immunotherapeutic strategies that harness the power of this phenomenon. While more investigations are needed to fully understand the mechanisms of trained immunity and its potential limitations, the prospects for its future application in clinical practice are promising. Here, we describe trained immunity as a biological process and explore the innate cues, epigenetic changes, and metabolic reprogramming activities that affect how trained immunity is induced.

ABSTRACT Avian leukosis is a significant viral disease affecting chicken populations globally, including Bangladesh, resulting in high mortality and morbidity rates and causing substantial economic losses in the commercial poultry industry. This study aimed to detect avian leukosis virus (ALV) during recent outbreaks in Bangladesh, utilising a molecular‐based approach. A total of 14 liver samples were collected from the suspected layer flocks in Bangladesh. The diagnosis of ALV infection in chickens was confirmed through necropsy, histopathological examinations, reverse transcription‐polymerase chain reaction (RT‐PCR), and sequence analysis. Gross observations revealed severe liver enlargement with scattered white nodules on the surface in the infected chickens. Histopathological observations showed the infiltration of huge mononuclear inflammatory cells in the periportal area of liver and microvesicular fatty degeneration and necrosis of some hepatocytes. RT‐PCR results identified three samples positive for the env gene of ALV. Sequence analysis of the env genes demonstrated high homology among the identified strains (97%–98%) and with reference strains (92%–96%) at the nucleotide level. The phylogenetic tree revealed close relatedness of the three identified strains to reference strains from India, USA, and China. Mutational analysis indicated several mutations throughout the envelope glycoprotein of the identified strains. Protein structure analysis showed minor changes in the hydrophobic region of the envelope protein of the identified strains. In conclusion, this study, the first detailed investigation in Bangladesh, contributes to understanding ALV epidemiology, highlights genetic diversity, and emphasises the necessity for further investigations and the implementation of effective control measures in the affected regions.

Antimicrobial resistance is a global threat. On the other hand, Bangladesh produces high-quality mangoes, yet the mango coat and seed kernel, which contain medicinal components, remain unused. Therefore, this study investigated the antibacterial potential and toxicity of ethanol extracts from mango (Mangifera indica) seed kernels, which specifically target the bacterial strains Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Klebsiella sp. Amrapali variant mango seeds were collected, dried, ground into a fine powder, and extracted with ethanol at various ratios. The efficacy of the crude extract was tested via the disc diffusion method. The results demonstrated significant antibacterial activity against gram-positive bacteria (S. aureus and B. cereus), with clear zones of inhibition observed, especially in a dose-dependent manner. The pure crude extract inhibited the growth of S. aureus with a zone of 23 mm, identical to that produced by doxycycline. However, the extract exhibited limited activity against gram-negative bacteria (E. coli and Klebsiella sp.). Additionally, the extract was effective against multidrug-resistant S. aureus. The pure crude extract produced a 22.5 mm zone of inhibition against multidrug-resistant S. aureus, which was slightly smaller than that of gentamicin (23 mm) but larger than those of chloramphenicol (21 mm), vancomycin (20 mm), and tetracycline (16 mm). In vivo toxicity was assessed in mice, revealing no significant adverse effects on the hepatic structure or renal cortex at lower doses (100 μl of pure crude extract). However, higher doses caused mild histopathological changes in the liver and kidneys. These findings suggest that mango seed kernel extract holds promise as an alternative antibacterial agent, particularly against gram-positive and antibiotic-resistant bacteria, while being relatively safe at lower doses. Further research is needed to elucidate the active compounds, mechanisms of action, and broader applications of this extract in combating antibiotic-resistant bacterial infections.

Abstract Introduction Lumpy skin disease, an economically significant bovine illness, is now found in previously unheard‐of geographic regions. Vaccination is one of the most important ways to stop its further spread. Aim Therefore, in this study, we applied advanced immunoinformatics approaches to design and develop an effective lumpy skin disease virus (LSDV) vaccine. Methods The membrane glycoprotein was selected for prediction of the different B‐ and T‐cell epitopes by using the immune epitope database. The selected B‐ and T‐cell epitopes were combined with the appropriate linkers and adjuvant resulted in a vaccine chimera construct. Bioinformatics tools were used to predict, refine and validate the 2D, 3D structures and for molecular docking with toll‐like receptor 4 using different servers. The constructed vaccine candidate was further processed on the basis of antigenicity, allergenicity, solubility, different physiochemical properties and molecular docking scores. Results The in silico immune simulation induced significant response for immune cells. In silico cloning and codon optimization were performed to express the vaccine candidate in Escherichia coli. This study highlights a good signal for the design of a peptide‐based LSDV vaccine. Conclusion Thus, the present findings may indicate that the engineered multi‐epitope vaccine is structurally stable and can induce a strong immune response, which should help in developing an effective vaccine towards controlling LSDV infection.

Adenoviruses are non-enveloped DNA viruses that cause a wide range of symptoms, from mild infections to life-threatening diseases in a broad range of hosts. Due to the unique characteristics of these viruses, they have also become a vehicle for gene-transfer and cancer therapeutic instruments. Adenovirus vectors can be used in gene therapy by modifying wild-type viruses to render them replication-defective. This makes it possible to swap out particular viral genes for segments that carry therapeutic genes and to employ the resultant vector as a means of delivering genes to specified tissues. In this review, we outline the progressive development of adenovirus vectors, exploring their characteristics, genetic modifications, and range of uses in clinical and preclinical settings. A significant emphasis is placed on their crucial role in advancing gene therapy, cancer therapy, immunotherapy, and the latest breakthroughs in vaccine development for various diseases.

This review delves into the historical context, current epidemiological landscape, genomics, and pathobiology of monkeypox virus (MPXV). Furthermore, it elucidates the present vaccination status and strategies to curb the spread of monkeypox. Monkeypox, caused by the Orthopoxvirus known as MPXV, is a zoonotic ailment. MPXV can be transmitted from person to person through respiratory droplets during prolonged face-to-face interactions. While many cases of monkeypox are self-limiting, vulnerable groups such as young children, pregnant women, and immunocompromised individuals may experience severe manifestations. Diagnosis predominantly relies on clinical presentations, complemented by laboratory techniques like RT-PCR. Although treatment is often not required, severe cases necessitate antiviral medications like tecovirimat, cidofovir, and brincidofovir. Vaccination, particularly using the smallpox vaccine, has proven instrumental in outbreak control, exhibiting an efficacy of at least 85% against mpox as evidenced by data from Africa. Mitigating transmission requires measures like wearing surgical masks, adequately covering skin lesions, and avoiding handling wild animals.

Antimicrobial resistance (AMR) is a crucial and emerging multifactorial "One Health" problem involving human and animal health, agriculture, aquaculture, and environment; and posing a potential public health hazard globally. The containment of AMR justifies effective surveillance programs to explicate the magnitude of the problem across the contributing sectors. Laboratory-based AMR testing and characterization is the key component of an AMR surveillance program. An AMR surveillance program should have a "top management" for fund mobilization, planning, formulating, and multilateral coordinating of the surveillance activities. The top management should identify competent participating laboratories to form a network comprising a reference laboratory and an adequate number of sentinel laboratories. The responsibilities of the reference laboratory include the development of standardized test methods for ensuring quality and homogeneity of surveillance activities, providing training to the laboratory personnel, and in-depth AMR characterization. The sentinel laboratories will take the responsibilities of receiving samples, isolation and identification of microbes, and initial AMR characterization. The sentinel laboratories will use simple antimicrobial susceptibility test (AST) methods such as disk diffusion tests, whereas the reference laboratories should use automated quantitative AST methods as well as advanced molecular methods to explicit AMR emergence mechanisms. Standard guidelines set by Clinical Laboratory Standards Institute or the European Committee on Antimicrobial Susceptibility Testing, should be followed to bring about conformity and harmonization in the AST procedures. AMR surveillance program in animals is eventually similar to that in human health with the exception is that veterinary antibiotics and veterinary pathogens should be given preference here. Hence, the review study was envisaged to look deep into the structure of the AMR surveillance program with significance on laboratory-based AMR testing and characterization methods.

Birds, especially wild waterfowl and migratory birds have the potential to carry antibiotic-resistant bacteria, but their role in the dissemination of these resistant pathogens is still neglected in Bangladesh. To the best of our knowledge, this study was carried out for the first time in Bangladesh to isolate and determine the occurrence of multidrug-resistant (MDR) Shigella spp. from fecal materials of wild waterfowl and migratory birds. A total of 80 fecal materials from wild waterfowl (n = 50) and migratory birds (n = 30) were screened to detect MDR Shigella isolates. Shigella spp. were isolated and identified by culturing, staining, and biochemical tests followed by polymerase chain reaction (PCR). A disk diffusion assay was employed to investigate antibiotic phenotypes, while the resistance genes were detected by PCR. Among the 80 samples, 15 (18.75%) were found positive for Shigella spp. by PCR, among which the occurrence rate of Shigella spp. was higher in migratory birds (20%, 6/30) than in wild waterfowl (18%, 9/50). By the disk diffusion test, 86.67% (13/15) of Shigella spp. isolates were found to be MDR in nature, including 93.33% of isolates resistant to imipenem. Moreover, frequent and moderate resistance was also observed against tetracycline (86.67%), azithromycin (80%), ampicillin (66.67%), ciprofloxacin and cotrimoxazole (40%), meropenem (26.67%), and streptomycin (13.33%). The bivariate analysis revealed a positive correlation between the resistance profiles of ciprofloxacin and cotrimoxazole, imipenem and tetracycline, tetracycline and ampicillin, and imipenem and azithromycin. Furthermore, the isolates had a multiple antibiotic resistance index of up to 0.47. Antibiotic resistance genes tetA and SHV were found in 69.23% and 50% of relevant antibiotic-resistant Shigella spp. isolates, respectively. The present study suggests that wild waterfowl and migratory birds are reservoirs of MDR Shigella spp., which may have detrimental impacts on One Health components. We suggest keeping these birds under an AMR monitoring program to avoid the possibility of AMR contamination of the environment and its consequences in all health settings.

Upland cotton is sensitive to 2,4-dichlorophenoxyacetic acid (2,4-D), and spray drift from adjacent crops can cause yield loss. The identification of potentially 2,4-D-tolerant cotton chromosome substitution (CS) lines with well-characterized tolerance mechanisms could provide a significant step into the development and genetic improvement of upland cotton. Experiments were conducted to understand the possible mechanism of herbicide tolerance in CS-T04-15, CS-T07, and CS-B15sh, 2,4-D herbicide-tolerant cotton CS lines compared with TM-1, the 2,4-D herbicide-susceptible recurrent parent of the CS line. Using [14C]2,4-D, the percent absorption rate and the translocation patterns of the 14C-labeled herbicide at 6 to 48 h after treatment (HAT) were determined. The tolerant cotton CS lines showed 15%–19% [14C]2,4-D uptake, while TM-1 exhibited a reduced uptake of only 1.4% [14C]2,4-D at 24 HAT. Distribution of the absorbed [14C]2,4-D showed that 2%–5% was translocated outside the treated leaf in the tolerant CS lines. In contrast, 77% of the herbicide was translocated above and below the treated leaf in line TM-1. Interestingly, CS-T04-15 showed a restricted movement of 14C below the treated leaf at 6 to 48 HAT, suggesting novel information on the mechanisms of herbicide tolerance to 2,4-D in cotton. This finding is the first report on upland cotton demonstrating a complex differential uptake and translocation associated with herbicide tolerance for [14C]2,4-D in cotton CS lines.

Enterococci are commensal bacteria that inhabit the digestive tracts of animals and humans. The transmission of antibiotic-resistant genes through human–animal contact poses a potential public health risk worldwide, as zoonoses from wildlife reservoirs can occur on every continent. The purpose of this study was to detect Enterococcus spp. in rhesus macaques (Macaca mulatta) and to investigate their resistance patterns, virulence profiles, and biofilm-forming ability. Conventional screening of rectal swabs (n = 67) from macaques was followed by polymerase chain reaction (PCR). The biofilm-forming enterococci were determined using the Congo red agar plate assay. Using the disk diffusion test (DDT), antibiogram profiles were determined, followed by resistance and virulence genes identification by PCR. PCR for bacterial species confirmation revealed that 65.7% (44/67) and 22.4% (15/67) of the samples tested positive for E. faecalis and E. faecium, respectively. All the isolated enterococci were biofilm formers. In the DDT, enterococcal isolates exhibited high to moderate resistance to penicillin, rifampin, ampicillin, erythromycin, vancomycin, and linezolid. In the PCR assays, the resistance gene blaTEM was detected in 61.4% (27/44) of E. faecalis and 60% (9/15) of E. faecium isolates. Interestingly, 88.63 % (39/44) of E. faecalis and 100% (15/15) of E. faecium isolates were phenotypically multidrug-resistant. Virulence genes (agg, fsrA, fsrB, fsrC, gelE, sprE, pil, and ace) were more frequent in E. faecalis compared to E. faecium; however, isolates of both Enterococcus spp. were found negative for the cyl gene. As far as we know, the present study has detected, for the first time in Bangladesh, the presence of virulence genes in MDR biofilm-forming enterococci isolated from rhesus macaques. The findings of this study suggest employing epidemiological surveillance along with the one-health approach to monitor these pathogens in wild animals in Bangladesh, which will aid in preventing their potential transmission to humans.

Objective: Chicken infectious anemia virus (CIAV) is an economically important emerging infec¬tion of poultry as it causes immunosuppression and reduces egg production. Although it is world¬wide distributed and first reported (single case) in Bangladesh in 2002, no epidemiological and serological investigations have been conducted. The current study aimed to conduct a serologi¬cal investigation on the prevalence of CIAV infection in broiler breeder and layer farms in some selected areas of Bangladesh. Materials and Methods: A total number of 460 sera samples were randomly collected from unvac¬cinated broiler breeder and layer flocks, of which 276 were from 11 broiler breeder farms and 184 from 12 layer farms. The sera samples were subjected to a commercially available enzyme-linked immunosorbent assay kit to observe antibodies induced by CIAV. Results: Results demonstrated that the overall prevalence of CIAV was 83.6% among a total of 460 samples. In broiler breeder birds, the prevalence was 89.9%, whereas it was 78.3% in layer birds. A higher number of female birds was found to be seropositive than male birds. However, chickens of all age groups were found to be susceptible to the virus. Conclusions: These results indicate the presence of CIAV in Bangladesh, which may be the sequel of naturally occurring either vertical or horizontal infection in all bird flocks tested without clinical symptoms of the disease. A further epidemiological investigation will be required, followed by molecular isolation and characterization of the virus for suitable vaccine candidate selection and/ or preparation. [J Adv Vet Anim Res 2021; 8(2.000): 323-329]

Coronavirus disease-19 (COVID-19) caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is now a pandemic threat. This virus is supposed to be spread by human to human transmission. Cellular angiotensin-converting enzyme 2 (ACE2) is the receptor of SARS-CoV-2 which is identical or similar in different species of animals such as pigs, ferrets, cats, orangutans, monkeys, and humans. Moreover, a recent study predicted that dogs might be secondary hosts during the evolution of SARS-CoV-2 from bat to human. Therefore, there is a possibility of spreading SARS-CoV-2 through domestic pets. There are now many reports of SARS-CoV-2 positive cases in dogs, cats, tigers, lion, and minks. Experimental data showed ferrets and cats are highly susceptible to SARS-CoV-2 as infected by virus inoculation and can transmit the virus directly or indirectly by droplets or airborne routes. Based on these natural infection reports and experimental data, whether the pets are responsible for SARS-CoV-2 spread to humans; needs to be deeply investigated. Humans showing clinical symptoms of respiratory infections have been undergoing for the COVID-19 diagnostic test but many infected people and few pets confirmed with SARS-CoV-2 remained asymptomatic. In this review, we summarize the natural cases of SARS-CoV-2 in animals with the latest researches conducted in this field. This review will be helpful to think insights of SARS-CoV-2 transmissions, spread, and demand for seroprevalence studies, especially in companion animals.

Background: Duck viral enteritis, commonly known as duck plague (DP), is an acute and contagious fatal disease in ducks, geese, and swan caused by the DP virus (DPV). It poses a serious threat to the growth of duck farming in the haor (wetland) areas of Bangladesh. Aim: This study aimed to detect circulating DPV by molecular characterization, followed by phylogenetic analysis, targeting the UL30 gene in infected ducks from five haor districts in Bangladesh and to observe the variation in the genome sequence between the field virus and vaccine strain of DPV. Methods: A total of 150 samples (liver, 50; intestine, 50; and oropharyngeal tissue, 50) were collected from DP-suspected sick/dead ducks from 50 affected farms in Kishoreganj, Netrokona, B. Baria, Habiganj, and Sunamganj districts of Bangladesh. For the identification of DPV in collected samples, polymerase chain reaction (PCR) was utilized. Nucleotide sequences of the amplified UL30 gene were compared with those of other DPV strains available in GenBank. Results: Of the 150 samples, 90 (60%) were found to be positive for DPV, as confirmed by PCR. Organ-wise prevalence was higher in the liver (72%), followed by the intestine (64%) and oropharyngeal tissue (44%). Regarding areas, the highest and lowest prevalence in the liver and intestine was observed in Habiganj and B. Baria, respectively, whereas the highest and lowest prevalence in the oropharyngeal tissue was observed in B. Baria and Habiganj, respectively. Two isolates, BAU/KA/DPV(B1)/2014 from Kishoreganj and BAU/KA/DPV(B4)/2014 from Sunamganj, were sequenced, and phylogenetic analysis revealed that these isolates are evolutionarily closely related to Chinese isolates of DPV. Additionally, the isolates of DPV BAU/KA/DPV(B1)/2014 and BAU/KA/DPV(B4)/2014 showed the highest (98%) similarity with each other. The nucleotide sequence of the isolate BAU/KA/DPV(B1)/2014 exhibited higher nucleotide variability (264 nucleotides) than that of the vaccine strain (accession no. EU082088), which may affect protein function and additional drug sensitivity. Conclusion: Based on the findings of the molecular study, it can be assumed that the Bangladeshi isolates and all Chinese isolates of DPV may have a common ancestry.

Antimicrobial resistance (AMR) is a crucial multifactorial and complex global problem and Bangladesh poses a regional and global threat with a high degree of antibiotic resistance. Although the routine application of antimicrobials in the livestock industry has largely contributed to the health and productivity, it correspondingly plays a significant role in the evolution of different pathogenic bacterial strains having multidrug resistance (MDR) properties. Bangladesh is implementing the National Action Plan (NAP) for containing AMR in human, animal, and environment sectors through "One Health" approach where the Department of Livestock Services (DLS) is the mandated body to implement NAP strategies in the animal health sector of the country. This review presents a "snapshot" of the predisposing factors, and current situations of AMR along with the weakness and strength of DLS to contain the problem in animal farming practices in Bangladesh. In the present review, resistance monitoring data and risk assessment identified several direct and/or indirect predisposing factors to be potentially associated with AMR development in the animal health sector of Bangladesh. The predisposing factors are inadequate veterinary healthcare, monitoring and regulatory services, intervention of excessive informal animal health service providers, and farmers' knowledge gap on drugs, and AMR which have resulted in the misuse and overuse of antibiotics, ultimate in the evolution of antibiotic-resistant bacteria and genes in all types of animal farming settings of Bangladesh. MDR bacteria with extreme resistance against antibiotics recommended to use in both animals and humans have been reported and been being a potential public health hazard in Bangladesh. Execution of extensive AMR surveillance in veterinary practices and awareness-building programs for stakeholders along with the strengthening of the capacity of DLS are recommended for effective containment of AMR emergence and dissemination in the animal health sector of Bangladesh.

Objectives: The present study estimated the seroprevalence of avian reovirus (ARV) infections in backyard chickens of the Mymensingh district in Bangladesh. Materials and Methods: Considering several risk factors, a total of 460 serum samples were col¬lected from backyard chickens from eight Upazilas of the Mymensingh district in Bangladesh. Blood samples were taken from the wing vein using 3-ml sterile syringes and kept at room tem¬perature for clotting in a slanting position and then transported to the laboratory maintaining the cool chain. Subsequently, the prepared sera were harvested and stored at −20°C until used. Finally, an indirect enzyme-linked immunosorbent assay (ELISA) was performed to detect ARV-specific antibodies using a commercial ARV antibody detection ELISA test kit. Results: The results revealed high prevalence rates of ARV antibodies, with a total seroprevalence of 69.78% (321/460). Area-wise, 74.55% (82/110) seroprevalence was recorded as the highest in Mymensingh Sadar, whereas 64% (32/50) was the lowest in Gauripur Upazila. With regard to sex, female chickens showed a significantly higher (p < 0.05) seroprevalence as 90.33% (271/300) compared to male chickens 31.25% (50/160). With regard to age groups, the seroprevalence of ARV infection was 59.33% (89/150) within 28 weeks, 82% (205/250) within 916 weeks, and 45% (27/60) within 1720 weeks, respectively. Based on hygienic conditions, the highest sero¬prevalence of ARV was noted in backyard chickens housed in poor conditions 80% (120/150) than good conditions 50% (40/80). Backyard chickens reared in free-ranging conditions exhibited a significantly higher seroprevalence 73.33% (220/300) of ARV antibodies compared to rearing in separate houses 63.12% (101/160). The seroprevalence of ARV was higher in crossbreeds 71.67% (43/60), brought from market 76% (38/50), and unhealthy 78.57% (55/70) backyard chickens than non-descriptive indigenous 69.5% (278/400), home-reared 69.02% (283/410), and healthy chick¬ens 68.21% (266/390). Conclusion: The high prevalence of ARV antibodies revealed in the current study indicates an extensive exposure of ARV to backyard chickens in Bangladesh that may be transmitted naturally to other chickens, ultimately leading to ominous economic effects on the poultry sector. [J Adv Vet Anim Res 2020; 7(3.000): 546-553]

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 phy¬logenetic analysis. Neighbor-joining trees were constructed by using the Molecular Evolutionary Genetics Analysis 6, having the field nucleotide sequences of FMDV and related sequences avail¬able 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% iden¬tity. A phylogenetic analysis revealed that the FMDV serotype O PanAsia-02 sub-lineage was con¬firmed in Bangladesh under the Middle EastSouth 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 neigh¬boring countries may act as the source of diversified FMDV serotypes in Bangladesh. [J Adv Vet Anim Res 2020; 7(2.000): 360-366]

Aim: Microbiological risk analysis of ready-to-eat fast foods for sale on the campus of Bangladesh Agricultural University was undertaken to understand the contribution of such foods to foodborne disease. Materials and Methods: From each of 10 randomly selected fast food outlets, a total of 120 samples of six food items were collected to assess common microbial load. In parallel, vendors were asked about their food production and selling practices, while consumers (n=200) were asked about their consumption of fast foods and whether they had experienced symptoms of foodborne disease. Results: Aerobic plate count (APC) varied from 6.92 to 7.24 log colony-forming unit (cfu)/g, Staphylococcus spp. 4.67 to 5.15 log cfu/g, Salmonella spp. 3.67 to 4.22 log cfu/g, and Escherichia coli 4.10 to 4.58 log cfu/g. Microbial risk assessment of Staphylococcus spp., Salmonella spp., and E. coli for daily, weekly, or monthly consumption was estimated by Monte Carlo simulation (10,000 iterations). The consumer survey showed 57% chance of being infected by ready-to-eat fast food samples. The calculated mean annual risks of Staphylococcus spp., Salmonella spp., and E. coli infection were about 100% in all cases. Conclusion: Thus, the study revealed high risk of infection associated with the consumption of ready-to-eat fast foods.

Objective: The study was aimed to analyze the microbiological quality of mixed vegetable salads and to understand the risk related with its consumption from different restaurants around Bangladesh Agricultural University campus in Mymensingh. Materials and Methods: Sixty (60) samples of mixed vegetable salads were taken from twelve (12) different restaurants in five different time points from each restaurant. In parallel, restaurant workers were asked for handling practices while the consumers were interviewed about their salad consumption pattern and whether they had experienced any health-related problems. Microbial risk assessment of Staphylococcus spp., Salmonella spp., and Escherichia coli was esti¬mated by Monte Carlo simulation (10,000 iterations), an exponential model. Results: Aerobic plate count was ranged from 7.73 ± 0.61 to 9.04 ± 0.26 log cfu/gm, Staphylococcus spp. from 4.64 ± 0.61 to 6.42 ± 0.53 log cfu/gm, Salmonella spp. from 4.75 ± 0.08 to 5.27 ± 0.53 log cfu/gm, and E. coli from 4.98 ± 0.20 to 6.66 ± 0.80 log cfu/gm. From the survey, it was found that total consumers had 18% chances where the male had 13% and the female had 30% chances of being infected with salads. Again frequent, average, and occasional consumers had 31%, 13%, and 0% chances, respectively, of being infected with those salads. From the Monte Carlo simulation, the calculated mean annual risks of Staphylococcus spp., Salmonella spp., and E. coli infection for the three exposure scenarios were found to be about 100%. Conclusion: The study actually revealed the potential microbial contamination in mixed vegetable salads which may impact on food safety and human health. So, the findings suggest that following hygienic measures during processing and handling the microbiological quality of mixed vegetables salads can be improved. [J Adv Vet Anim Res 2020; 7(1.000): 34-41]

Background and Aim: Houseflies (Musca domestica) are synanthropic insects which serve as biological or mechanical vectors for spreading multidrug-resistant bacteria responsible for many infectious diseases. This study aimed to detect antibiotic-resistant bacteria from houseflies, and to examine their resistance genes. Materials and Methods: A total of 140 houseflies were captured using sterile nylon net from seven places of Mymensingh city, Bangladesh. Immediately after collection, flies were transferred to a sterile zipper bag and brought to microbiology laboratory within 1 h. Three bacterial species were isolated from houseflies, based on cultural and molecular tests. After that, the isolates were subjected to antimicrobial susceptibility testing against commonly used antibiotics, by the disk diffusion method. Finally, the detection of antibiotic resistance genes tetA, tetB, mcr-3, mecA, and mecC was performed by a polymerase chain reaction. Results: The most common isolates were Staphylococcus aureus (78.6%), Salmonella spp., (66.4%), and Escherichia coli (51.4%). These species of bacteria were recovered from 78.3% of isolates from the Mymensingh Medical College Hospital areas. Most of the isolates of the three bacterial species were resistant to erythromycin, tetracycline, penicillin and amoxicillin and were sensitive to ciprofloxacin, ceftriaxone, chloramphenicol, gentamicin, and azithromycin. Five antibiotic resistance genes of three bacteria were detected: tetA, tetB, mcr-3, and mecA were found in 37%, 20%, 20%, and 14% isolates, respectively, and no isolates were positive for mecC gene. Conclusion: S. aureus, Salmonella spp., and E. coli with genetically-mediated multiple antibiotic resistance are carried in houseflies in the Mymensingh region. Flies may, therefore, represent an important means of transmission of these antibiotic-resistant bacteria, with consequent risks to human and animal health.

Infectious bursal disease (IBD) is a highly contagious disease that causes significant economic loss in chickens. A cross-sectional study was conducted in the Mymensingh district of Bangladesh to determine the seroprevalence of IBD virus (IBDV) antibodies in backyard chickens and their association with different epidemiological risk factors. A total of 460 serum samples were randomly collected from backyard chickens that had not been previously vaccinated against IBDV. The collected sera were examined using an enzyme-linked immunosorbent assay (ELISA). Data on epidemiological risk factors were collected through face-to-face interviews with owners and subjected to both uni- and multivariable risk analyses to determine their association with IBDV infection. Using ELISA, the overall seroprevalence of IBDV antibodies in backyard chickens was 83.4% (95% confidence interval: 79.8%–86.6%), among which, a significantly higher seroprevalence was recorded in females (83.4%, 345/350), 4–6 weeks age group (95.3%, 244/256), and unhealthy (95.0%, 57/60) backyard chickens than those of males, other age groups, and healthy chickens, respectively. Furthermore, chickens reared in free-ranging housing systems (93.3%, 280/300) and poor-conditioned houses (98.0%, 147/150) showed a significantly higher seropositivity of IBDV antibodies than those reared in separated housing systems and other hygienic-conditioned houses, respectively. Moreover, compared with their counterparts, a higher but nonsignificant seroprevalence of IBDV antibodies was observed in backyard chickens that were selected from Fulbaria Upazila (88.8%; 80/90) and which were brought from the marketplace (85.7%, 60/70). A higher seropositivity of IBDV antibodies was shown to be statistically associated with various critical epidemiological risk factors, indicating that field strains of IBDV were exposed in backyard chickens and could be readily transferred horizontally. Proper prevention and control methods, villagers’ awareness of IBD, and the rapid and widespread use of seroepidemiological investigations could help to reduce the spread of IBDV infection in backyard chickens.

Cotton genetic resources contain diverse economically important traits that can be used widely in breeding approaches to create of high-yielding elite cultivars with superior fiber quality and adapted to biotic and abiotic stresses. Nevertheless, the creation of new cultivars using conventional breeding methods is limited by the cost and proved to be time consuming process, also requires a space to make field observations and measurements. Decoding genomes of cotton species greatly facilitated generating large-scale high-throughput DNA markers and identification of QTLs that allows confirmation of candidate genes, and use them in marker-assisted selection (MAS)-based breeding programs. With the advances of quantitative trait loci (QTL) mapping and genome-wide-association study approaches, DNA markers associated with valuable traits significantly accelerate breeding processes by replacing the selection with a phenotype to the selection at the DNA or gene level. In this review, we discuss the evolution and genetic diversity of cotton Gossypium genus, molecular markers and their types, genetic mapping and QTL analysis, application, and perspectives of MAS-based approaches in cotton breeding.

The subtype prevalence, drug resistance- and pathogenicity-associated mutations, and the distribution of the influenza A virus (IAV) isolates identified in Bangladesh from 2002 to 2019 were analyzed using bioinformatic tools. A total of 30 IAV subtypes have been identified in humans (4), avian species (29), and environment (5) in Bangladesh. The predominant subtypes in human and avian species are H1N1/H3N2 and H5N1/H9N2, respectively. However, the subtypes H5N1/H9N2 infecting humans and H3N2/H1N1 infecting avian species have also been identified. Among the avian species, the maximum number of subtypes (27) have been identified in ducks. A 3.56% of the isolates showed neuraminidase inhibitor (NAI) resistance with a prevalence of 8.50, 1.33, and 2.67% in avian species, humans, and the environment, respectively, the following mutations were detected: V116A, I117V, D198N, I223R, S247N, H275Y, and N295S. Prevalence of adamantane-resistant IAVs was 100, 50, and 30.54% in humans, the environment, and avian species, respectively, the subtypes H3N2, H1N1, H9N2, and H5N2 were highly prevalent, with the subtype H5N1 showing a comparatively lower prevalence. Important PB2 mutations such D9N, K526R, A588V, A588I, G590S, Q591R, E627K, K702R, and S714R were identified. A wide range of IAV subtypes have been identified in Bangladesh with a diversified genetic variation in the NA, M2, and PB2 proteins providing drug resistance and enhanced pathogenicity. This study provides a detailed analysis of the subtypes, and the host range of the IAV isolates and the genetic variations related to their proteins, which may aid in the prevention, treatment, and control of IAV infections in Bangladesh, and would serve as a basis for future investigations.

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]

Abstract Background Brucellosis is a zoonotic disease caused by bacteria Brucella spp. belonging to the genus Brucella. It is endemic in domesticated animals in Bangladesh. Isolation, identification and genetic characterization of Brucella spp. in dairy cattle are essential to undertake appropriate control and preventive measures. The study was conducted to isolate and characterize the Brucella spp. circulating in dairy cattle. Methods Uterine discharge (n = 45), milk (n = 115), vaginal swab (n = 71), placenta (n = 7) and aborted fetus (n = 2) were collected. Brucella selective agar plates were inoculated with samples and incubated at 37 ◦C for 14 days under 5% CO2 for isolation of Brucella spp. Brucella suspected colonies were recovered from samples were confirmed by genus and species specific PCR assays. Genetic characterization was performed by Multi Locus Variable number tandem‐repeat Analysis‐16 (MLVA‐16). Results The isolates of Brucella recovered from samples were confirmed as B. abortus by AMOS‐ERY PCR assay. The classical biotyping method confirmed all 10 B. abortus isolates belonged to the biovar 3. The MLVA‐16 assay indicated all B. abortus isolates identical and the same genotype 40, based on panel 1 MLVA‐8. Conclusion Dendrogram analysis revealed all B. abortus isolates of the study were identical to three isolates from Brazil, one isolate of France and closely related to Chinese isolates. This is the first report of isolation and genetic characterization of B. abortus from the dairy cattle in Bangladesh.

Objective: The study was aimed to isolate, identify, and characterize common indicator bacteria, including Escherichia coli, Salmonella spp., and Staphylococcus spp. in manure and bio-slurry sam¬ples of different livestock farms and biogas plants of Bangladesh. Materials and Methods: A total of 114 samples of manure and bio-slurry were collected from different livestock farms and biogas plants in Bangladesh. The total viable count (TVC), E. coli, Salmonella spp., and Staphylococcus spp. counts were determined by the spread plate technique method. Isolation and identification were performed by colony characteristics, staining, bio¬chemical tests, and, finally, by using PCR. Antibiotic susceptibility test of the isolated bacteria was tested against commonly used antibiotics by using the disk diffusion method. Results: The mean TVC, E. coli, Salmonella spp., and Staphylococcus spp. counts were ranged from 8.1910.75, 5.26.96, 5.816.87, 5.687.68 in manure samples and 7.268.65, 3.825.2, 45.54, 3.145.9 log cfu/gm in bio-slurry, respectively. In anaerobic digester after 30 days digestion, the presence of E. coli, Salmonella spp., and Staphylococcus spp. varied from 05.11, 04.84, and 05.59 log cfu/gm at 25°C, 27°C, 29°C, and 45°C temperature. Above-mentioned bacteria were absent in bio-slurry collected from anaerobic digester after 60 days digestion at environmental temperature. Bacterial counts were reduced significantly in both household slurry pits and exper¬imental anaerobic digester. Antibiotic susceptibility results revealed that multidrug-resistant indi¬cator bacteria were present in the bio-slurry samples. Conclusion: Our findings conclude that the microbial load after treatment of animal manure via anaerobic digestion (Biogas plant) was grossly reduced and the reduction of bacterial pathogen depends on the duration and temperature of digestion. [J Adv Vet Anim Res 2019; 6(3.000): 376-383]

Background and Aim: Avian reovirus (ARV) is a constraint to poultry industry in Bangladesh as a cause of several diseases in chickens, especially in broiler. However, the actual status of the viral infection is not known because the large-scale study is not conducted in this country. Therefore, this study aimed to check the presence and distribution of ARV-specific antibody in respect to area, types of chickens (broiler breeder, broiler, and layer), vaccination status, and age of chickens in Gazipur and Mymensingh districts of Bangladesh. Materials and Methods: A total of 276 chickens' blood samples were collected from two well-organized broiler breeder stock, seven broiler farms, and five layer farms located at two districts, namely Gazipur and Mymensingh of Bangladesh. Blood samples were collected from wing vein of the apparently healthy chickens using 3 ml of syringe and serum was harvested by keeping the syringe at room temperature in slanting position. The sera were transferred to the laboratory by maintaining the cool chain and further processing was performed by indirect enzyme-linked immunosorbent assay using ARV antibody test kit. Results: The results of serological test revealed that an average of 39.5% seropositive against ARV was recorded in chickens of Gazipur and Mymensingh districts. Among these, chickens of Gazipur district had the highest seropositivity of 50.5% than Mymensingh (30.7%). With respect to vaccination status, the seropositivity of vaccinated chickens in both areas was 100% and non-vaccinated chickens was 50.5% in Gazipur and 30.7% in Mymensingh district, respectively. However, regarding age groups, the seropositivity was higher in the age of 4-6 weeks (64.5%). Conclusion: The present serological findings showed a higher prevalence of ARV-specific antibodies in broiler birds. It indicates that the poultry industries of Bangladesh are contaminated with ARV which may naturally be transmitted to chickens either vertically or horizontally.

Objectives: This study was conducted to assess the prevalence and characterization of Staphylococcus aureus from chicken and quail eggshells and to study the antibiogram of the isolates. Materials and methods: A total of 300 eggs (220 chicken eggs and 80 quail eggs) were collected from different retail shops and farms in Mymensingh district. Swabs taken from the egg surfaces were cultured on Mannitol Salt Agar for the isolation of S. aureus. Polymerase chain reaction was conducted for confirmatory identification of the bacterial species targeting nuc gene, followed by confirmation of methicillin-resistant S. aureus by targeting the mecA gene. Antibiotic sensitivity test of the isolated bacteria was done against commonly used antibiotics by the disk diffusion method. Results: The prevalence of Staphylococcus spp. and S. aureus in the chicken eggshell surface was 20.45% and 10.45%, respectively. Similarly, the prevalence of Staphylococcus spp. and S. aureus in quail eggshell surface was 16.25% and 5%, respectively. Overall, 27 isolates were identified as S. aureus, of which 23 were from the chicken eggshell surface and four from quail eggshell surface. Among the seven isolates tested, overall four (57.14%) were positive for the nuc gene. On the other hand, the mecA gene could be detected in three (50%) S. aureus out of six oxacillin resistant isolates. The antibiogram study indicated that most of the isolates were resistant to the antibiotics under β-lactam group. Conclusion: The present study concludes that chicken and quail egg surface harbor multidrugresistant bacteria which may cause public health hazards, if these antibiotic-resistant bacteria are transferred to a human. [J Adv Vet Anim Res 2018; 5(4.000): 466-471]

Objective: The research work was conducted to determine the duration of protective efficacy and lowest immune protective titer of Salmonella bivalent vaccine containing Salmonella gallinarum and Salmonella pullorum prepared at the Livestock and Poultry Vaccine Research and Production Centre (LPVRPC) of Bangladesh Agricultural University (BAU), Mymensingh, Bangladesh. Materials and methods: The experimental chickens were subdivided into four main groups (A, B, C and D). Group A and B were vaccinated with BAU-Salmonella bivalent vaccine with dosed 0.5 mL intramuscularly at the age of seven weeks followed by a booster vaccination at 12 weeks of age while group C and D served as unvaccinated control. The sera samples were obtained at 7, 12, 15, 18, 23, 27, 30, 32, 34, 36 and 41 weeks of age of birds. Results: Significantly elevated level of immune response in terms of antibody production resulted from booster vaccination. Vaccinated chicken showed protective resistance following virulent challenge with isolates of S. gallinarum and S. pullorum (**P [J Adv Vet Anim Res 2018; 5(3.000): 282-289]

Objective: A cross sectional study was conducted in five Haor (wetland) districts of Bangladesh to investigate the epidemiological parameters and associated factors of recurrent outbreak of duck plague (DP) on the basis of molecular detection. Materials and methods: A total of 450 randomly selected duck farms containing 175,467 ducks were investigated for their morbidity, mortality and case fatality rates from December 2012 to November 2014. The farms were surveyed and the owners were interviewed using a prepared questionnaire about husbandry practice, disease history, bio-security measures, and flock and farmer details. A total of 150 samples (liver=50, intestine=50 and oro-pharyngeal tissue=50) were collected from duck plague suspected sick/dead ducks of north eastern part of Bangladesh during outbreak season. Samples were processed and PCR was done directly from the samples using primers specific for duck plague virus. Results: Out of 150 test samples of the fifty duck plague suspected ducks of fifty outbreaks, 90 samples (60%) of 36 ducks of thirty six outbreaks (72%) were found positive by PCR. Overall prevalence of duck plague was 8% at farm level and 3.30% at flock level. Morbidity, mortality and case fatality rates of duck plague at farm level were 52.08, 29.62 and 56.86%, respectively. Of the 22 variables selected for this study, 16 were found significant and the remaining 6 were found non-significant statistically. Conclusion: Results of the epidemiological investigation of the present study regarding duck mortality suggesting that education and training of the farmers on bio-security, modern husbandry practice, regular vaccination and innovation of cost effective intensive duck farming methods are necessary to control recurrent duck plague outbreak in Haor (wetland) areas. [J Adv Vet Anim Res 2018; 5(2.000): 131-139]

Palmer amaranth is a problematic common weed species, especially in cotton. With the wide use of chemical herbicide and herbicide-tolerant transgenic cotton lines, Palmer amaranth populations have developed tolerance to commonly used herbicides. It is imperative to develop alternative weed control methods to slow the evolution of herbicide-resistant weed populations and provide new strategies for weed management. Eleven chromosome substitution (CS) cotton lines (CS-B26lo, CS-T17, CS-B16-15, CS-B17-11, CS-B12, CS-T05sh, CS-T26lo, CS-T11sh, CS-M11sh, CS-B22sh, and CS-B22lo) were screened for weed-suppressing abilities in this study. The cotton lines were tested using the established stair-step assay. Height (cm) and chlorophyll concentration (cci) were measured for each plant in the system. The most significant variation in Palmer amaranth height reduction among the CS lines was observed 21 days after establishment. CS-B22sh (76.82%) and T26lo (68.32%) were most effective in reducing Palmer amaranth height. The cluster analysis revealed that CS-B22sh, and CS-T26lo were clustered in one group, suggesting similar genetic potential with reference to Palmer amaranth growth and development. CS-B22sh showed novel genetic potential to control the growth and development of Palmer amaranth, a problematic weed in cotton fields. Future experimentation should implement more parameters and chemical testing to explore allelopathic interactions among CS lines and Palmer amaranth.

Nutrients, including macronutrients such as Ca, P, K, and Mg, are essential for crop production and seed quality, and for human and animal nutrition and health. Macronutrient deficiencies in soil lead to poor crop nutritional qualities and a low level of macronutrients in cottonseed meal-based products, leading to malnutrition. Therefore, the discovery of novel germplasm with a high level of macronutrients or significant variability in the macronutrient content of crop seeds is critical. To our knowledge, there is no information available on the effects of chromosome or chromosome arm substitution on cottonseed macronutrient content. The objective of this study was to evaluate the effects of chromosome or chromosome arm substitution on the variability and content of the cottonseed macronutrients Ca, K, Mg, N, P, and S in chromosome substitution lines (CS). Nine chromosome substitution lines were grown in two-field experiments at two locations in 2013 in South Carolina, USA, and in 2014 in Mississippi, USA. The controls used were TM-1, the recurrent parent of the CS line, and the cultivar AM UA48. The results showed major variability in macronutrients among CS lines and between CS lines and controls. For example, in South Carolina, the mean values showed that five CS lines (CS-T02, CS-T04, CS-T08sh, CS-B02, and CS-B04) had higher Ca level in seed than controls. Ca levels in these CS lines varied from 1.88 to 2.63 g kg−1 compared with 1.81 and 1.72 g kg−1 for TM-1 and AMUA48, respectively, with CS-T04 having the highest Ca concentration. CS-M08sh exhibited the highest K concentration (14.50 g kg−1), an increase of 29% and 49% over TM-1 and AM UA48, respectively. Other CS lines had higher Mg, P, and S than the controls. A similar trend was found at the MS location. This research demonstrated that chromosome substitution resulted in higher seed macronutrients in some CS lines, and these CS lines with a higher content of macronutrients can be used as a genetic tool towards the identification of desired seed nutrition traits. Also, the CS lines with higher desired macronutrients can be used as parents to breed for improved nutritional quality in Upland cotton, Gossypium hirsutum L., through improvement by the interspecific introgression of desired seed nutrient traits such as Ca, K, P, S, and N. The positive and significant (p ≤ 0.0001) correlation of P with Ca, P with Mg, S with P, and S with N will aid in understanding the relationships between nutrients to improve the fertilizer management program and maintain higher cottonseed nutrient content.

Upland cotton (Gossypium hirsutum L.) growth and development during the pre-and post-flowering stages are susceptible to high temperature and drought. We report the field-based characterization of multiple morpho-physiological and reproductive stress resilience traits in 11 interspecific chromosome substitution (CS) lines isogenic to each other and the inbred G. hirsutum line TM-1. Significant genetic variability was detected (p < 0.001) in multiple traits in CS lines carrying chromosomes and chromosome segments from CS-B (G. barbadense) and CS-T (G. tomentosum). Line CS-T15sh had a positive effect on photosynthesis (13%), stomatal conductance (33%), and transpiration (24%), and a canopy 6.8 °C cooler than TM-1. The average pollen germination was approximately 8% greater among the CS-B than CS-T lines. Based on the stress response index, three CS lines are identified as heat- and drought-tolerant (CS-T07, CS-B15sh, and CS-B18). The three lines demonstrated enhanced photosynthesis (14%), stomatal conductance (29%), transpiration (13%), and pollen germination (23.6%) compared to TM-1 under field conditions, i.e., traits that would expectedly enhance performance in stressful environments. The generated phenotypic data and stress-tolerance indices on novel CS lines, along with phenotypic methods, would help in developing new cultivars with improved resilience to the effects of global warming.

Micronutrients are essential for plant growth and development, and important for human health nutrition and livestock feed. Therefore, the discovery of novel germplasm with significant variability or higher micronutrients content in crop seeds is critical. Currently, there is no information available on the effects of chromosome or chromosome arm substitution in cotton on cottonseed micronutrients. Thus, the objective of this study was to evaluate the effects of chromosome or chromosome arm substitution on the variability and levels of micronutrients B, Fe, Cu, Zn, Mn, and Ni in cottonseed from chromosome substitution (CS) cotton lines. Our hypothesis was that interspecific chromosome substitution in cotton can affect cottonseed micronutrients content, resulting in significant differences and variabilities of these nutrients among CS lines and between CS lines and the controls. Nine CS lines were grown in two-field experiments at two locations (in 2013 in South Carolina, USA; and in 2014 in Mississippi, USA). TM-1 (the recurrent parent of the CS line) and AM UA48 (cultivar) were used as control. The results showed significant variability among CS lines compared to the controls AM UA48 and TM-1. For example, in South Carolina (SC), B concentration in cottonseed ranged from 10.35 mg kg−1 in CS-M02 to 13.67 mg kg−1 in CS-T04. The concentration of Cu ranged from 4.81 mg kg−1 in CS-B08sh to 7.65 mg kg−1 in CS-T02, and CS-T02 was higher than both controls. The concentration of Fe ranged from 36.09 mg kg−1 to 56.69 mg kg−1 (an increase up to 57%), and six CS lines (CS-B02, CS-B08sh, CS-M02, CS-M04, CS-T02, and CS-T04) had higher concentration than both controls in 2013. In 2014 at the Mississippi location (MS), similar observation was found with CS lines for micronutrients content. The CS lines with higher concentrations of these micronutrients can be used as a genetic tool toward QTL identification for desired seed traits because these lines are genetically similar with TM-1, except the substituted chromosome or chromosome segment pairs from the alien species. Chromosome substitution provides an effective means for upland cotton improvement by targeted interspecific introgression, yielding CS lines that facilitate trait discovery, such as seed micronutritional qualities, due to increased isogenicity and markedly reduced complexity from epistatic interactions with non-target alien chromosomes. The positive correlation between B, Cu, and Fe at both locations, between Ni and Mn, between Zn and Cu, and between Zn and Ni at both locations signify the importance of a good agricultural and fertilizer management of these nutrients to maintain higher cottonseed nutrient content.

Brucellosis is endemic in Bangladesh both in humans and in animals. A number of reasons complicate the diagnosis, as bovine brucellosis can be diagnosed by various serological tests. But the tests have a limitation; when the organism remains intracellular, the disease goes into chronic stage and the antibody titres may decline. The present study was conducted for isolation and detection of Brucella spp. by polymerase chain reaction (PCR) from seronegative cows. A total of 360 dairy cows from three geographical regions were screened serologically by Rose Bengal Plate Test (RBPT) where 24 samples were serologically positive and the rest of the samples were serologically negative. Among the 24 seropositive individuals, 11 were culture positive and 6 were culture positive from serologically negative dairy cows. The overall seroprevalence of brucellosis in cattle was 6.6% and in disease condition a higher prevalence was recorded in abortion (28.07%) followed by infertility (13.33%). To confirm the Brucella spp. in seronegative dairy cattle, the isolates were extracted and PCR was conducted, which produced 905 bp amplicon size of 6 Brucella spp. from milk sample. So, for the detection or eradication of brucellosis, a bacteriological test and a PCR technique should be performed with the serological test of milk.

Cotton fiber is an important commodity throughout the world. Fiber property determines fiber quality and commercial values. Previous studies showed that silencing phytochrome A1 gene (PHYA1) by RNA interference in Upland cotton (Gossypium hirsutum L. cv. Coker 312) had generated PHYA1 RNAi lines with simultaneous improvements in fiber quality (longer, stronger and finer fiber) and other key agronomic traits. Characterization of the altered molecular processes in these RNAi genotypes and its wild-type controls is a great interest to better understand the PHYA1 RNAi phenotypes. In this study, a total of 77 conserved miRNAs belonging to 61 families were examined in a PHYA1 RNAi line and its parental Coker 312 genotype by using multiplex sequencing. Of these miRNAs, seven (miR7503, miR7514, miR399c, miR399d, miR160, miR169b, and miR2950) were found to be differentially expressed in PHYA1 RNAi cotton. The target genes of these differentially expressed miRNAs were involved in the metabolism and signaling pathways of phytohormones, which included Gibberellin, Auxin and Abscisic Acid. The expression of several MYB transcription factors was also affected by miRNAs in RNAi cotton. In addition, 35 novel miRNAs (novel miR1-novel miR35) were identified in fibers for the first time in this study. Target genes of vast majority of these novel miRNAs were also predicted. Of these, nine novel miRNAs (novel-miR1, 2, 16, 19, 26, 27, 28, 31 and 32) were targeted to cytochrome P450-like TATA box binding protein (TBP). The qRT-PCR confirmed expression levels of several differentially regulated miRNAs. Expression patterns of four miRNAs-targets pairs were also examined via RNA deep sequencing. Together, the results imply that the regulation of miRNA expression might confer to the phenotype of the PHYA1 RNAi line(s) with improved fiber quality.

Most wild and semi-wild species of the genus Gossypium are exhibit photoperiod-sensitive flowering. The wild germplasm cotton is a valuable source of genes for genetic improvement of modern cotton cultivars. A bi-parental cotton population segregating for photoperiodic flowering was developed by crossing a photoperiod insensitive irradiation mutant line with its pre-mutagenesis photoperiodic wild-type G. darwinii Watt genotype. Individuals from the F2 and F3 generations were grown with their parental lines and F1 hybrid progeny in the long day and short night summer condition (natural day-length) of Uzbekistan to evaluate photoperiod sensitivity, i.e., flowering-time during the seasons 2008-2009. Through genotyping the individuals of this bi-parental population segregating for flowering-time, linkage maps were constructed using 212 simple-sequence repeat (SSR) and three cleaved amplified polymorphic sequence (CAPS) markers. Six QTLs directly associated with flowering-time and photoperiodic flowering were discovered in the F2 population, whereas eight QTLs were identified in the F3 population. Two QTLs controlling photoperiodic flowering and duration of flowering were common in both populations. In silico annotations of the flanking DNA sequences of mapped SSRs from sequenced cotton (G. hirsutum L.) genome database has identified several potential 'candidate' genes that are known to be associated with regulation of flowering characteristics of plants. The outcome of this research will expand our understanding of the genetic and molecular mechanisms of photoperiodic flowering. Identified markers should be useful for marker-assisted selection in cotton breeding to improve early flowering characteristics.

Dispersion of Love waves is studied in a fibre-reinforced layer resting on monoclinic half-space. The wave velocity equation has been obtained for a fiber-reinforced layer resting on monoclinic half space. Shear wave velocity ratio curve for Love waves has been shown graphically for fibre reinforced material layer resting on various monoclinic half-spaces. In a similar way, shear wave velocity ratio curve for Love waves has been plotted for an isotropic layer resting on various monoclinic half-spaces. From these curves, it has been observed that the curves are of similar type for a fibre reinforced layer resting on monoclinic half-spaces, and the shear wave velocity ratio ranges from 1.14 to 7.19, whereas for the case isotropic layer, this range varies from 1.0 to 2.19.