Objective: This study was carried out to estimate the sex ratio, maturity size, gonadosomatic index (GSI), and peak breeding season of mud crabs.  Materials and Methods: Samples were collected randomly from the estuary and river of the study area. Sampling was carried out monthly from April to September at every full moon during one high tide. A total number of 240 specimens were sampled, where 53 individuals were hermaphro­dite. The crabs were shifted alive to the biology and histology lab for detailed biological study. Sex was determined. Male and female sex ratio and breeding season were also investigated.  Results: The male:female ratio was 1:0.96 and the ovarian development was categorized into five stages based on internal observations, viz. immature (stage I), underdeveloped (stage II), early developed (stage III), late developed (stage IV), and mature (stage V). The maturity percentages were 37%, 19%, 13%, 11%, and 20%, respectively. 50% maturation was estimated at 82.36 mm inter­nal carapace width (ICW). The highest mean GSI value was 7.97 ± 3.03. The mature stage was found in all the working periods. This shows that females have activated ovaries in all the working months, and the species are continuous breeders. A higher frequency of vitellogenic ovary and higher GSI value were found in September. The maximum GSI value was found in the size group 70–79 mm.  Conclusion: The study shows that the capture from the wild sources of mud crabs without any regulation can threaten the population structure. The capture of female mud crabs should be more than 82.36 mm ICW, which will help conserve and protect young crabs. J. Adv. Vet. Anim. Res., 8(1): 44-50, Mar 2021 http://doi.org/10.5455/javar.2021.h483

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 circu­lating 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., 8(1): 51-57, Mar 2021 http://doi.org/10.5455/javar.2021.h484

Objective: The paper’s primary goal is to report the devastating impact of carbon tetrachloride (CCl4) on rat testicular tissue and the possible protecting function of propolis against CCl4 based on its free radical scavenging and inflammatory relief properties.  Materials and Methods: A total of 24 adult male albino rats had been classified into four groups (six rats/group). Rats of group 1 served as control, whereas groups 2–4 received propolis (200 mg/kg/day), CCl4 (3 ml/kg/day), and propolis/CCl4, respectively. After 4 weeks, the collected sera were applied for the estimation of lipid profile and sex hormones. Also, histopathological picture, malondialdehyde, and tumor necrosis factor alpha (TNFα) gene profile was measured in collected testicular tissues.  Results: The present information revealed a noteworthy change (p < 0.05) in lipid profile, decrease in testicular weight, testosterone, antioxidants values along with a prominent increase (p < 0.05) in estradiol, lipid peroxidation values, and expression of TNFα in rats administrated with CCl4 com­pared to control. Moreover, the histopathological profile showed the degeneration of the epithe­lium. Interestingly, propolis attenuated the destructive effect of CCl4 on rat testes.  Conclusion: The examined dose of propolis reduced oxidation, and inflammatory reactions resulted from CCl4 exposure and proved that it might have a helpful part in free radicals inter­ceded diseases. J. Adv. Vet. Anim. Res., 8(1): 70-77, Mar 2021 http://doi.org/10.5455/javar.2021.h487

Objectives: Demodicosis is a vital skin problem in dogs. The present study has determined the prevalence and associated risk factors of demodicosis in dogs and the response to treatment.  Materials and Methods: A total of 100 skin scrapings were collected from dogs having dermatolog­ical lesions brought to the Teaching and Training Pet Hospital and Research Center of Chattogram Veterinary and Animal Sciences University for treatment purpose. The collected scrapings were dissolved in 10% potassium hydroxide to detect mites through microscopic examination. Various risk factors like breed, age, sex, hair type, health status, and management system (indoor and outdoor) were analyzed using the logistic regression model. Positive cases were treated with oral ivermectin (Scabo®; at 0.6 mg/kg/day) along with amitraz 12.5% (Ridd®) diluted to 0.05% for rub­bing on the body after bathing with chlorhexidine shampoo (PetHex®). Clavulanate amoxicillin (Moxaclav®) and omega-3 fatty acids (OMG-3®) were also suggested to prevent secondary bacte­rial infection and to maintain skin and hair coat integrity. The recovery rate was observed every 2–4 weeks of treatment upto 11–13 weeks.  Results: The overall prevalence of demodicosis was recorded as 27%. Hair type, health status, and management system were significantly (p < 0.05) associated with the occurrence of demodicosis in dogs. Following treatment, the first negative skin scraping for mite was noticed at 8–10 weeks of treatment, and in all cases, clinical signs completely disappeared at 80–90 days of treatment.  Conclusion: A good percentage of the dogs having dermatological lesions was determined as demodicosis. Long-term oral ivermectin and topical amitraz, an oral antibiotic, and nutritional therapy are effective against canine demodicosis. Keeping long-haired dogs with good body con­dition and indoor management is highly suggestive of preventing and controlling the disease. J. Adv. Vet. Anim. Res., 8(1): 116-122, Mar 2021 http://doi.org/10.5455/javar.2021.h493

Objective: This experiment was undertaken to assess the scope and cost-effectiveness of the fer­mented corn mixture (FCM)-based buffalo fattening approach compared to urea molasses straw (UMS) and silage-based approach.  Materials and Methods: A completely comparative randomized design experiment was con­ducted for 90 days with three treatments and five buffalo bulls in each. UMS, silage, and FCM roughage-based fattening diets were attributed as T1, T2, and T3, respectively. Two types of protein supplements, i.e., Type 1 (Bangladesh Livestock Research Institute-developed) for T1 and T2 and Type 2 (prescribed by farmers) for T3 treatments, were used. All the parameters were analyzed through Statistical Package for the Social Sciences, 20 software.  Results: Dry matter intake (kg, %live weight) was significantly higher in T1 (2.65), followed by T2 (2.34) and T3 (2.00), respectively. The crude protein intake, digestible crude protein intake, and digestible dry matter intake (kg/d) significantly (p < 0.05) differed between T1 and T3, but not T2. The digest­ibility of acid detergent fiber (65.97) was significantly higher for T3 than T1 and T2 (54.44 and 58.73, respectively). Neutral detergent fiber digestibility of T3 (70.35) also differed (p < 0.05) with T1 (60.97) but not T2 (64.78). No difference was observed in the case of growth, but feed conversion ration was found to be significantly (p < 0.05) better in T2 (7.10) than T1 (8.35), where T3 (7.24) was neutral. The significantly (p < 0.001) highest expense [216.37 Bangladesh taka (BDT)/kg gain] was required for T1, followed by T2 and T3 (174.47 and 126.33 BDT/kg gain, respectively). Net profit from T3 and T2 (15,877 and 15,175 BDT, respectively) gained significantly (p < 0.05) higher than T1 (11,265 BDT).  Conclusion: The FCM-based diet was suitable and cost-effective as a buffalo fattening approach. J. Adv. Vet. Anim. Res., 8(2): 195-202, June 2021

Objective: The objective of this study was to determine the location, number, and direction of the nutrient foramen in the humerus, radius, femur, and tibia bones of mixed breed dogs.  Materials and Methods: The humerus, radius, femur, and tibia of both (left and right) limbs of mixed breed dogs were examined in this study. The number, location, and direction of the nutrient foramina were identified. Once identified, the diameter of each nutrient foramen was measured and the site index calculated.  Results: Only one nutrient foramen was identified in the humerus, radius, tibia, and right femur, while the foramen numbers ranged from one to three in the left femurs examinated. The nutri­ent foramen was localized on the caudal surface in the radii, femurs, tibias, and left humeri. Contrasting, however, 75% were located on the caudal surface of the right humeri and 25% on the lateral surface. The average diameter of the nutrient foramen of the humerus ranged from 0.88 to 1.00 mm, while it ranged from 1.13 to 1.25 mm in the radius. On the hind limb, the diameter of the nutrient foramen on the femur ranged from 1.2 to 1.3 mm and 0.75–1.25 mm on the tibia. The nutrient foramen was directed towards the corresponding joint in 100% of the humeri and tibias, 75% of the radii, and 60%–80% of the femurs examined.  Conclusion: The anatomical data on the nutrient foramen obtained in this study will be valuable for veterinarians when diagnosing pathological bone lesions and for orthopedic surgery. J. Adv. Vet. Anim. Res., 8(2): 203-209, June 2021

Objective: Sertoli cells (SCs) are important sustentacular cells in the seminiferous tubules of the testis. Isolation and identification of SCs are the premise for studying their functions. Since New Zealand rabbit is a stable strain which is widely used for biomedical research and animal farming, this study aimed to develop a simple and effective protocol for SC isolation in New Zealand rabbits.  Materials and Methods: The SCs of three 30-day-old New Zealand rabbits were isolated by incu­bation with enzymatic digestion I (Dulbecco’s modified Eagle medium supplemented with 1 mg/ ml collagenase IV and 50 μg/ml DNase I) and digestion II (digestion I + 1 mg/ml hyaluronidase + 1 mg/ml trypsin), as well as differential plating. The cells were enriched and identified by using immunocytochemical staining and reverse transcription polymerase chain reaction analysis.  Results: Homogeneous cells were obtained. They presented the typical large cell body and an irregular pyramidal shape after differential plating and passaging. These cells expressed mRNA of the SC marker sex-determining region Y-box 9 (SOX9) instead of the Leydig cell marker StAR. Immunocytochemically, they are positive of SOX9, GATA binding protein 4, and androgen-binding protein.  Conclusion: The SCs were enriched from the testicular tissues of prepubertal New Zealand rabbits by a simple and effective protocol, which provides a basis for further theoretical researches and practical applications. J. Adv. Vet. Anim. Res., 8(2): 218-223, June 2021

Objective: Previously, we have shown that predicted zymogen granule protein 16 homolog B (P-G3MZ19) existed in Bali cattle (Bos javanicus) saliva. It was suggested that P-G3MZ19 is a mem­ber of the mannose-binding lectin family that plays an essential role in innate immunity. In the present study, we aimed to analyze the structure and ligand-binding of P-3MZ19 in Bali cattle saliva.  Materials and Methods: Saliva of four adult healthy Bali cattle was collected, lyophilized, and subjected to two-dimensional (2-D) gel electrophoresis. The target spot of around 17 kDa related to P-G3MZ19 was excised for matrix-assisted laser desorption ionization time-of-flight mass spec­trometer/time-of-flight mass spectrometer mass spectrometry analysis and sequencing. The structure and the ligand-binding of P-3MZ19 were analyzed using bioinformatics software pro­grams published elsewhere.  Results: Based on Iterative Threading ASSEmbly Refinement the 3D model of P-G3MZ19 was suggested to have similarities to exo-alpha-sialidase (EC 3.2.1.18); while its ligand-binding sites consisted of seven residues, i.e., 25aa-26aa (Gly-Gly), 95aa (Phe), 138aa (Tyr), 140aa (Leu), 141aa (Gly), and 143aa (Thr).  Conclusion: The structure of P-G3MZ19 of Bali cattle saliva and its ligand-binding sites have been successfully determined by using bioinformatics techniques. The biological and immunological roles of the peptide are currently under investigation based on P-G3MZ19 synthetic peptides. J. Adv. Vet. Anim. Res., 8(2): 224-229, June 2021

Objective: The TYR (Tyrosinase) and MC1R (Melanocortin 1 receptor) genes are recognized as important genes involved in plumage pigmentation in Korean native chickens. Specifically, most color patterns in chicken result from differential expression of the TYR gene. In this study, the co-segregation of the pigmentation and sequence of the TYR and MC1R genes was investigated through intercrosses between red (R1q1), red with black and black plumage color types of native Korean chickens.  Materials and Methods: Using DNA, RNA, and tissue by plumage color of each Korean native chickens, the role of major genes in pigmentation of pheomelanin was evaluated. Reverse tran­scription polymerase chain reaction, sequencing, western blot, and immunohistochemical were performed to determine the effect of TYR and MC1R genes on plumage pigmentation in Korean native chickens.  Results: The KCO line (Korean chicken Ogol: Black-line) with an EEC _ genotype exhibited black feathers, whereas red and red mixed with black chicken with EeC genotype exhibited white feathers. There were notable differences between the base sequences of MC1R and TYR in three Korean chicken breeds, with the highest variation in TYR. Perhaps this is the key characteristics of Korean chicken. Further, we analyzed the expression patterns of MC1R and TYR genes in each type of Korea native chicken and observed that TYR expression was high in feather follicle (R1q2) of KCO tissue. However, native red (Korean chicken red) and native red with black (Korean chicken red dark) chickens have increased TYR expression in the tissue. However, the expression of MC1R was much different from that of TYR.  Conclusion: In this study, our results suggest that the differences in position and TYR expres­sion levels exert more influence on plumage pigmentation in native Korean chicken breeds than changes in MC1R expression levels. J. Adv. Vet. Anim. Res., 8(2): 266-273, June 2021

Objective: Fast foods are often responsible for staphylococcal foodborne illness. The present study was carried out to isolate Staphylococcus spp. from various fast foods sold in Mymensingh and to determine their antibiogram.  Materials and Methods: Overall, 60 samples of fast foods sold in different restaurants were screened by culture, biochemical tests, and polymerase chain reaction (PCR) to isolate and iden­tify Staphylococcus spp., followed by employing of disk diffusion method to determine their anti­biotic resistance patterns.  Results: Among these 60 samples, 8 [13.33%, 95% confidence interval (CI): 6.91%–24.17%] were positive for Staphylococcus spp. by cultural and biochemical properties. By PCR, four (6.67%, 95% CI: 2.62%–15.93%) isolates were confirmed as Staphylococcus aureus by targeting nuc gene. Additionally, all the S. aureus isolates were coagulase-positive. By antibiogram profiles, all the Staphylococcus isolates exhibited resistance to azithromycin and erythromycin (95% CI: 67.56%– 100.00%), and frequently resistance to cefixime (75%, 95% CI: 40.93%–95.56%), ampicillin (50%, 95% CI: 21.52%–78.48%), and amoxicillin (37.5%, 95% CI: 13.68%–69.43%); moderate to lower resistance was found against cefotaxime, gentamicin, and doxycycline. In addition, all the isolates were sensitive to ciprofloxacin and chloramphenicol. Interestingly, 75% (6/8; 95% CI: 40.93%– 95.56%) isolates were multidrug-resistant (MDR) in nature. Furthermore, the indices of multiple antibiotic resistance (MAR) were ranged from 0.2 to 0.6.  Conclusion: This study revealed that fast foods sold in restaurants were contaminated with MDR and MAR Staphylococcus isolates, having potential public health significance. J. Adv. Vet. Anim. Res., 8(2): 274-281, June 2021