Objective: The objective of this study was to evaluate the effects of caffeine and taurine on the motility and viability of chilled equine semen. Materials and Methods: A total of 12 ejaculates were collected from three mature stallions with proven fertility during the breeding season. The gel-free spermatic fraction of each ejaculate was divided into two aliquots and diluted with a semen extender (either INRA 96® or BotuSemen Gold®). The aliquots were then split and assigned to one of the six treatment groups: control (no supplement), caffeine (2 and 4 mM), taurine (25 and 50 mM), and a combination of caffeine (2 mM) plus taurine (25 mM). Samples were stored at 4°C and analyzed at different time points (0, 24, 48, 72, and 96 h) to evaluate total (TMOT) and progressive (PMOT) motility and viability by computer-assisted sperm analysis. Results: Regardless of the extender, PMOT and TMOT decreased over time. However, compared with the control, the treatment with 4 mM caffeine significantly mitigated the decrease in PMOT at 72 h. Additionally, semen treated with a combination of caffeine plus taurine maintained a significantly higher PMOT at 96 h, with improved viability at all time points. Conclusions: The combination of caffeine plus taurine helps maintain chilled equine semen viabil¬ity and progressive motility up to 96 h independently of the extender used. [J Adv Vet Anim Res 2021; 8(4.000): 635-641]

Objective: The study investigated the effect of different stocking density (SD) rates on the welfare, growth, and hemato-biochemical parameters in broiler chickens. Materials and Methods: 106 broiler chicks of 10 days old were used and assigned into four groups: A, B, C, and D. The chicks of group A were reared in floor space containing one bird per square foot area (SD1.0). The chicks of groups B, C, and D were reared at 1.5, 2.0, and 2.5 birds per square foot area (SD1.5, SD2.0, and SD2.5). Welfare, body weight, and hemato-biochemical parameters were assessed and monitored by physical observation and laboratory methods. Results: The birds reared at SD2.0, and SD2.5 rates showed increased panting breathing. Wet feces adhered below the vent. There were a significant number of birds showing dirtiness of body and feathers. Birds reared in SD2.5 were familiar with moist litters and high ammonia smell. Foot-pad dermatitis, scratches, and blister formation were detected in the leg. The study revealed that the higher SD negatively correlated to the welfare behavior indicators. Live body weight was significantly (p < 0.05) decreased in birds reared at higher SD rates. Birds housed in SD1.0 and SD1.5 are optimum for body weight and improved feed conversion ratio. The hemato-biochemical parameters of birds reared at various SD rates did not differ. The total leucocyte count increased significantly, while total serum proteins decreased gradually as SD rates increased. Conclusion: This work explores that higher SD negatively affects welfare and growth performance in broiler chickens. [J Adv Vet Anim Res 2021; 8(4.000): 642-649]

Objective: Duck virus enteritis is a severe viral disease that kills ducks and swans worldwide. The clinical manifestations, gross pathology, molecular detection, and characterization of the duck virus enteritis virus (DVEV) in Australian black swans at a safari park in Bangladesh were described in this case report. Materials and Methods: On a safari park in Bangladesh, an Australian black swan flock exhibited clinical signs of anorexia, greenish watery diarrhea, increased thirst, partial paralysis, and death. Postmortem examinations of deceased swans revealed extensive pathological abnormalities in the trachea, liver, and spleen. To isolate DVEV, a viral inoculum produced from the liver and spleen of dead swans was implanted into 913-day-old embryonated duck eggs via the chorioallantoic membrane (CAM) route. DVEV was confirmed using a polymerase chain reaction (PCR) assay. Phylogenetic analysis was used to determine the genetic relationship between the DVEV isolates from Australian black swans, and 16 DVEV isolates previously described in the GenBank. Results: Hemorrhage was noted in the annular ring of the trachea, as well as an enlarged and hemorrhagic liver and spleen. The PCR assay amplified a 446-bp fragment of the DVEV DNA polymerase gene in the liver, spleen, and CAM homogenates. The phylogenetic analysis found that the DVEV isolates from swans were comparable to those from Bangladesh, India, Vietnam, China, Germany, the USA, and Egypt. Conclusion: According to the findings of this study, the DVEV was the cause of illness and mortality in an Australian black swan flock. [J Adv Vet Anim Res 2021; 8(4.000): 557-562]

Objective: The article aimed to study the relationship between the physico-chemical qualities of pig meat and their pre-slaughter weights. Materials and Methods: In this study, 60 pigs were reared for fattening under the same condi¬tions of keeping and feeding, slaughtered, and used to sample the longest back muscle meat with pre-slaughter weights of 110 and 130 kg. The samples were evaluated according to generally accepted methods for assessing the physico-chemical qualities in pig meat. Results: Samples of meat from animals slaughtered at 130 kg had higher values for marbling by 2.0 points or 5.24% (p < 0.01), active acidity pH in ham muscles by 0.20 pH or 3.57% (p < 0.01), and the longest muscle of the back by 0.10 pH or 1.82% (p < 0.001). The pre-slaughter weight factor substantially affected the pH of ham muscles at 10.35% and on the marbling of meat in the longest back muscle at 13.31%. Pigs slaughtered at 110 kg had a greater increase in the color intensity of the meat and an increase in its water holding capacity. At a pre-slaughter weight of 130 kg, increasing the marbling and decreasing the softness of the flesh demonstrated a modest, adverse relationship. Conclusion: The findings support the use of pre-slaughter weight management to enhance pork quality. [J Adv Vet Anim Res 2021; 8(3.000): 521-533]

Objectives: This study aims to investigate the prevalence and antibiogram of Staphylococcus aureus and methicillin-resistance S. aureus (MRSA) in rabbits, rabbit handlers, and rabbitry environments in Terengganu. Materials and Methods: Swab samples from 183 rabbits (183 oral and 183 ear swabs), 45 rabbit handlers (45 oral and 45 nasal), and environmental (n = 180) samples from rabbitries were collected from 10 rabbit farms in Terengganu. The associated S. aureus isolates from the swabs were isolated using phenotypic microbiology tests. The bacteria were confirmed by polymerase chain reaction targeting nuc (S. aureus) and mecA (MRSA) genes. The antibiogram of all S. aureus isolates was determined using the KirbyBauer test. Results: Staphylococcus aureus was detected in 19% of rabbits, 26.7% of rabbit handlers, and 8.8% of swabs from the rabbitry environment. However, MRSA (0%) could not be detected. Antibiotic susceptibility test revealed that S. aureus from rabbits showed low resistance ( [J Adv Vet Anim Res 2021; 8(3.000): 388-395]

Objective: Natural substances found in dietary sources and medicinal plants have attracted con¬siderable attention in recent years as chemopreventive agents. Spirulina is a blue-green alga that possesses chemopreventive properties. The purpose of this study was to determine the effect of spirulina on rats with inorganic arsenic (As) [sodium arsenite (NaAsO2)]-induced lipid peroxidation. Materials and Methods: 120 rats were randomly assigned to 10 groups and designated T0, T1, T2, T3, T4, T5, T6, T7, T8, and T9. One group was kept as a control (T0) that received no treatment. The seven groups received 3.0 mg of NaAsO2/kg body weight in drinking water and were given spirulina ad libitum. T1 was treated with NaAsO2 but not with spirulina. Two groups of rats (T2 and T3), on the other hand, were treated with spirulina without receiving any As (NaAsO2). T2 received agro-based spirulina (Ab-Sp; grown in 1.5% soybean meal media and harvested on day 12 of seed inoculation) at 2.0 gm/kg feed, whereas T3 received commercially available spirulina (Com-Sp) at 2.0 gm/kg feed. T4, T5, and T6 were concurrently treated with Ab-Sp at 1.0, 1.5, and 2.0 gm/ kg of feed. On the other hand, T7, T8, and T9 induced by NaAsO2 were concurrently treated with Com-Sp at 1.0, 1.5, and 2.0 gm/kg feed. All groups received treatment for 90 days. Results: The efficacy of both spirulina in preventing lipid peroxidation caused by As was deter¬mined quantitatively by measuring the rats serum malondialdehyde (MDA). The results indicated that As supplementation increased serum MDA levels, whereas both types of spirulina signifi¬cantly decreased them. The highest dose of Ab-Sp (2.0 gm/kg feed) was found to be the most effective in preventing lipid peroxidation in rats treated with inorganic As. Conclusion: Ab-Sp could be a natural, cost-effective, and safe measure to mitigate As toxicity. [J Adv Vet Anim Res 2021; 8(2.000): 330-338]

Objective: Humpback (hpbk) mice harbor a pathogenic mutation in the Notch3 gene and can serve as a beneficial animal model for investigating human myopathy, kyphosis, and developmen¬tal disorders, including lateral meningocele syndrome. Detection of the point mutation in hpbk mice is important for maintaining strains and scrutinizing genetic rescues, especially considering that homozygous mice are infertile and indistinguishable from their littermates at a young age. This study aimed for the development of a novel, precise, and time-saving genotyping method to identify the mutation in hpbk mice. Materials and Methods: In order to study the hpbk mouse line, we describe how we applied sev¬eral tools, including quantitative polymerase chain reaction (qPCR), multiplex tetra-primer ampli¬fication-refractory mutation system (ARMS-PCR) and Sanger sequencing, toward the recognition of heterozygous and homozygous mice. Results: The Notch3 mutation was clearly identified using qPCR and ARMS assays, but the latter was a more precise and cost-effective approach. The lengths of the ARMS-PCR amplicons are 210 bp and 164 bp for the wild-type and hpbk alleles, respectively. Moreover, the genotyping results for each mouse were corroborated by Sanger DNA sequencing. Conclusion: Our newly developed PCR-based ARMS system affords a swift and precise way to genotype the hpbk mice. ARMS-PCR does not rely on any advanced equipment and is useful as a genotyping method for other model organisms that harbor a pathogenic variant. [J Adv Vet Anim Res 2021; 8(1.000): 7-13]

Objectives: The comprehensive in silico study aims to figure out the most effective aromatic phytochemical ligands among a number from a library, considering their pharmacokinetic effi¬cacies in blocking angiotensin-converting enzyme 2 (ACE2) receptorsevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) S protein complex formation as part of a target-specific drug designing. Materials and Methods: A library of 57 aromatic pharmacophore phytochemical ligands was prepared from where the top five ligands depending on Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) and quantitative structure-activity relationship (QSAR)-based pharmacokinetic properties were considered. The selected ligands were optimized for commenc¬ing molecular docking and dynamic simulation as a complex with the ACE2 receptor to compare their blocking efficacy with the control drug. The ligandreceptor complexes accuracy in prevent¬ing the Spike (S) protein of SARS-CoV-2 penetration inside the host cells has been analyzed through hydrogenhydrophobic bond interactions, principal component analysis (PCA), root mean square deviation (RMSD), root mean square fluctuation (RMSF), and B-Factor. Advanced in silico program¬ming language and bioanalytical software were used for high throughput and authentic results. Results: ADMET and QSAR revealed Rhamnetin, Lactupicrin, Rhinacanthin D, Flemiflavanone D, and Exiguaflavanone A as the ligands of our interest to be compared with the control Cassiarin D. According to the molecular docking binding affinity to block ACE2 receptor, the efficiency mount¬ings were Rhinacanthin D > Flemiflavanone D > Lactupicrin > Exiguaflavanone A > Rhamnetin. The binding affinity of the Cassiarin DACE2 complex was (−10.2 KJ/mol) found inferior to the Rhinacanthin DACE2 complex (−10.8 KJ/mol), referring to Rhinacanthin D as a more stable candi¬date to use as drugs. The RMSD values of proteinligand complexes evaluated according to their structural conformation and stable binding pose ranged between 0.1~2.1 Å. The B-factor showed that very few loops were present in the protein structure. The RMSF peak fluctuation regions ranged 5250, predicting efficient ligandreceptor interactions. Conclusion: The experiment sequentially measures all the parameters required in referring to any pharmacophore as a drug, considering which all aromatic components analyzed in the study can strongly be predicted as target-specific medication against the novel coronavirus 2019 infection. [J Adv Vet Anim Res 2021; 8(1.000): 24-35]

Objective: Avian influenza is a zoonotic disease with a pandemic potential that can infect avian and mammalian species, including humans. Studies aimed at investigating avian influenza virus (AIV) status in asymptomatic chickens and their shedding are uncommon in Bangladesh. Therefore, the current study aimed to examine the distribution of AIV subtypes in asymptomatic commercial chicken flocks and to identify the possible risk factors associated with this infection in two selected sub-districts of Bangladesh. Materials and Methods: A total of 582 oropharyngeal swabs were collected from 23 chicken farms during 2019 and evaluated for the presence of AIV and its subtypes by real-time reverse transcription PCR assays. Risk factors associated with AIV infection were analyzed from questionnaire data. Results: Overall, AIV prevalence was 7.73% (n = 45) with 7.39% and 7.92% in Dhamrai and Gazipur Sadar sub-districts, respectively. In AIV-positive samples, the prevalence of A/H5N1, A/H5N2, A/ H9N1, and A/H9N2 was 31.11%, 28.89%, 6.67%, and 8.89%, respectively. None of the samples were positive for N6 and N8. The odds ratio (OR) of AIV infection was 1.15 in broiler versus layer and 2 in Sonali versus layer chickens. The OR was 1.95 for medium versus small, 2.6 for large versus small flock size, 1.5 for moderate versus good biosecurity, and 2.92 for poor versus good biosecurity practicing farms. Conclusion: The results demonstrated that A/H5N1, A/H5N2, A/H9N1, and A/H9N2 are circu¬lating in asymptomatic chickens of selected areas. Strict farm biosecurity practices and avoiding higher flock density are recommended to prevent AIV spread in the study. [J Adv Vet Anim Res 2021; 8(1.000): 51-57]

Objective: The purpose of this work was to investigate the combination of early-age acclimation and linseed dietary inclusion in enriching polyunsaturated fatty acids (PUFAs) in broilers meat as a strategy to mitigate heat stress. Materials and Methods: A total of 400 broiler chicks were assigned to four experimental groups with four duplicates (25 animals each): C: control (basal diet), AC: early-age acclimated (basal diet), Cl: fed 5% ground linseed, and Acl: early-age acclimated and fed 5% ground linseed. The lipid and fatty acid contents of different parts (breast, thigh, liver, subcutaneous, and abdominal fat) of broilers were determined. Results: Low levels of lipids and unsaturated fatty acids have been found in the meat of accli¬mated broilers. Higher levels of linolenic acids were noted in Cl thigh meat compared to C (6% vs. 2.68%, respectively). The results showed that oleic and linoleic acids constitute a large part of the PUFAs of different meats. The most elevated levels of monounsaturated fatty acids were recorded in the breast meat of AcL animals. The highest content of omega-3 was recorded in the liver of AcL animals compared to that of C (14.98% vs. 7.8%, respectively). Conclusion: We suggest that the combination of treatments during hot conditions has led to the reversion of the environment-affected variables to accepted values, and yields better ther¬moresistance, PUFA-enriched meat, and safeguard animal health which conferred to birds better solutions to reduce fatigue and hypoxic activities, which induces a considerable consumption of oxygen. [J Adv Vet Anim Res 2021; 8(2.000): 237-245]