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: Collagen is a fibrous protein that is primarily used in the pharmaceutical and biomedical industries. This study isolates and characterizes type-1 collagen from Sardine longiceps (scales, skin, and muscle). Materials and Methods: Collagen was isolated from S. longiceps using two methods: acid-solubilized collagen and pepsin-solubilized collagen. Sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDSPAGE) was used to estimate the molecular weight of isolated collagen. Ultraviolet (UV)-visible spectrophotometry analysis was used to confirm the collagen extracted (type-I collagen). The functional groups of isolated collagens were identified using fourier trans¬form infrared (FTIR) analysis. The X-ray diffraction (XRD) technique was used to investigate the crystallinity of isolated collagen. The high-pressure liquid chromatography (HPLC) technique was used to study the amino acid composition. Results: SDSPAGE of S. longiceps revealed molecular weights ranging from 116 kDa for α-2 to 97 kDa for α-1. UV-visible spectra showed an absorbance value below 300 nm, and the results confirmed type-I collagen. FTIR showed major functional groups like amide A, B, I, II, and III. XRD determined the crystallinity of isolated collagen. The HPLC results showed the presence of higher glycine content, followed by proline and hydroxyl proline in the extracted collagen. Conclusion: The overall study confirmed that fish waste materials (scales, skin, and muscles) could be used as an alternative source for collagen. [J Adv Vet Anim Res 2021; 8(4.000): 679-686]

Objective: The purpose of this study was to investigate how pomegranate peel extract (PPE) can prevent lipid oxidation, peroxide value, and pathogenic bacteria growth in buffalo meat. Materials and Methods: PPE and buffalo meat were employed in this investigation. The buffalo meat marinated with PPE was evaluated by refrigerating it at a temperature of 5°C ± 1°C on days 0, 4, 8, 12, and 16. PPE was added to buffalo meat at a rate of 0% as a control (K0), 0.50% (K1), 1.00% (K2), 1.50% (K3), and 2.00% (K4). Results: The addition of PPE lowered the total plate count, peroxide value, lipid, and pH between treatments and storage period (p < 0.05). PPEs high concentration of polyphenols, flavonoids, antioxidants, and antibacterial substances may decrease lipid oxidation, peroxide production, and bacterial growth rate. Conclusions: Marinating buffalo meat in PPE may help maintain the meats freshness while being stored at a refrigerator temperature (5°C ± 1°C). [J Adv Vet Anim Res 2021; 8(4.000): 612-618]

Objective: This study aimed to isolate and identify Escherichia coli from broiler samples from Sukabumi, Indonesia. Also, antibiogram studies of the isolated bacteria were carried out consid¬ering the detection of the antibiotic resistance genes. Materials and Methods: Cloaca swabs (n = 45) were collected from broilers in Sukabumi, Indonesia. Isolation and identification of E. coli were carried out according to standard bacterio¬logical techniques and biochemical tests, followed by confirmation of the polymerase chain reac¬tion targeting the uspA gene. Antibiotic sensitivity test, using several antibiotics [tetracycline (TE), oxytetracycline (OT), ampicillin (AMP), gentamicin (CN), nalidixic acid (NA), ciprofloxacin (CIP), enrofloxacin (ENR), chloramphenicol, and erythromycin] was carried out following the Kirby Bauer disk diffusion method. Detection of antibiotic resistance coding genes was carried out by PCR using specific oligonucleotide primers. Statistical analysis was carried out with one-way anal¬ysis of variance. Results: The results showed that 55.6% (25/45) of the samples were associated with the pres¬ence of E. coli. Antibiotic sensitivity test showed that the E. coli isolates were resistant to TE (88%; 22/25), OT (88%; 22/25), AMP (100%; 25/25), CN (64%; 16/25), NA (100%; 22/25), CIP (88%; 22/25), ENR (72%; 18/25), chloramphenicol (0%; 0/25), and erythromycin (92%; 23/25). On the other hand, the antibiotic resistance coding genes were tetA (86.4%; 19/22), blaTEM (100%; 25/25), aac(3)-IV (0%; 0/16), gyrA (100%; 25/25), and ermB (13%; 3/23). It was found that chlor¬amphenicol is markedly different from other antibiotic treatment groups. Conclusion: Escherichia coli was successfully isolated from cloacal swabs of broiler in Sukabumi, Indonesia. The bacteria were resistant to TE, OT, AMP, CN, NA, CIP, ENR, and erythromycin. Chloramphenicol was more sensitive and effective than other antibiotics in inhibiting the growth of E. coli. The antibiotic resistance genes detected were tetA, blaTEM, gyrA, and ermB. [J Adv Vet Anim Res 2021; 8(1.000): 84-90]

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 nutri¬ent 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 0.751.25 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 2021; 8(2.000): 203-209]

Objective: The objective of this study was to obtain an insight into the role of the vascular endothelial growth factor (VEGF) in pregnancy-associated mammary epithelial development and angiogenesis. However, we examined the primary VEGF receptor (VEGFR-2) in the mouse mammary cycle. Materials and Methods: The virgin (1012 weeks), days 10 and 18 of pregnancy (P-10 and P-18), days 0, 5, 10, and 20 of lactation (L-0, L-5, L-10, and L-20), and days 5 and 10 of post-weaning (W-5 and W-10) stage were all used in this study. Immunohistochemistry and Western blotting were carried out on mammary tissues taken from three mice at each stage. Results: VEGFR-2 was detected immunohistochemically in the cytoplasm of mammary epithelial and endothelial cells. Immunostaining for VEGFR-2 was consistently positive in mammary endothelial cells across all stages, whereas staining intensity in epithelial cells varied across the mammary cycle. Additionally, immunoblot analysis indicated a 220 kDa unique band of VEGFR-2 protein at all stages of the mammary cycle, with the maximum expression reported toward the end of pregnancy and gradually decreasing toward the end of lactation. Conclusion: In conclusion, the presence of VEGFR-2 in the mammary epithelium in addition to the endothelium suggests that VEGF plays an autocrine and paracrine role in the development, proliferation, and differentiation of the mammary epithelium during pregnancy. [J Adv Vet Anim Res 2021; 8(4.000): 581-588]

Objective: The present study aimed to explain the effect of fetal bovine serum (FBS) on the in vitro production of porcine embryos and the molecular effects of FBS on the growing of porcine embryos. Materials and Methods: Immature porcine oocytes were matured and fertilized in vitro. The resulting zygotes were cultured in porcine zygotic medium-3- until day 7 and FBS was added on day 4. Without FBS, it was treated as a control group. Quantitative real-time PCR and 2′,7′-dichloro-di¬hydro-fluorescein diacetate (H2DCFDA) molecular staining techniques were used to detect the expression patterns of apoptosis-associated genes and the accumulation of reactive oxygen species (ROS), respectively. Paired students t-test was used by GraphPad Prism statistical software. Results: FBS supplementation boosted blastocyst (BL) development and total cell count per BL substantially (p < 0.05). However, hatching and hatched BLs also increased in the FBS-treated group compared to the control. We also found that ROS accumulation in FBS-treated embryos was significantly reduced (p < 0.05) compared to the control group. The expression of the anti-apoptotic gene BCL-2 was significantly increased in FBS-treated BLs, but the pro-apoptotic gene, caspase-3 expression, was significantly reduced in FBS-treated BLs. Conclusion: Our results suggest that FBS supplementation in porcine culture media could increase porcine embryo production by decreasing ROS accumulation and increasing the anti-apoptotic gene expression in developing BLs. [J Adv Vet Anim Res 2021; 8(4.000): 589-596]

Objectives: The purpose of this study was to determine the species composition of Eimeria circu¬lating in Mymensingh district, Bangladesh, using Internal Transcribed Spacer 1 (ITS1) sequences in polymerase chain reaction (PCR) assay. Materials and Methods: Coccidian oocysts were isolated and sporulated in a solution containing 2% potassium dichromate from litter slurry collected from 13 commercially active broiler farms in the research region. Genomic DNA was isolated from sporulated oocysts and used to amplify the Eimeria species-specific ITS1 gene by PCR amplification. Electrophoresis of 1.5% agarose gel was used to visualize the amplified PCR products. Results: In the study samples from Mymensingh district, Bangladesh, the presence of Eimeria brunetti, Eimeria acervulina, Eimeria necatrix, Eimeria mitis, and Eimeria tenella was identified. Conclusions: The findings of this study may shed light on the zonal approach to chicken coccidio¬sis control. Additionally, it suggests that ITS1-based PCR might be used in the field to accurately identify Eimeria species. [J Adv Vet Anim Res 2021; 8(3.000): 489-493]

Vitamin C supplementation is important for the growth and development of bullfrog tadpoles under optimum water temperature conditions. Therefore, an experiment was carried out to evaluate the effects of vitamin C supplementation on the diet of bullfrog tadpoles at a low temperature. A total of 480 tadpoles with a mean weight of 0.078 g were distributed in 12 aquariums each containing 40 L of water in a closed water recirculation system. The experimental design was entirely randomized with four treatments (0, 150, 300, and 600 mg kg-1 of L-ascorbic acid monophosphate) and three replicates. The productive performance was measured by the weight gain, feed conversion, diet consumption, protein efficiency, carcass yield, hepatosomatic index, viscerosomatic index, visceral fat index, dry matter, and ethereal carcass extract. The water temperature during the experimental period was 21.74 ± 0.43 °C. Vitamin C supplementation did not influence carcass yield and viscerosomatic index. However, there was a quadratic effect of vitamin C supplementation on the weight gain, apparent feed conversion, protein efficiency, visceral fat index, hepatosomatic index, and ethereal carcass extract. Based on these results, bullfrog tadpoles should be supplemented with 600 mg vitamin C kg-1 of the diet when subjected to water temperatures of around 22 °C.

The objective of this study was to determine the ability of prostaglandin E2 (PGE2) to induce ovulation and expression of PGE2 receptor (EP2 and EP4) and COX genes (COX-1 and COX-2) in the ovary and pituitary of prepubertal mice. The positive control consisted of the application of 5 μg of gonadotropin-releasing hormone (GnRH, n = 29); the negative control applied 0.5 mL of phosphate buffered saline (PBS, n=31); the treatment tested the application of 250 μg of PGE2 (n = 29), making a total of 89 prepubertal mice (BALB/c). Mice were euthanized 14 to 15 h after treatments to detect ovulation and tissue collection. A Chi-square test was used to compare the proportion of animals ovulating. Gene expressions and number of ovulation were analyzed by one-way ANOVA and Tukey’s test was used to compare means among groups. A greater proportion of mice (P < 0.001) ovulated after receiving GnRH (89.7%, 26/29) compared to PGE2 group (58.6%, 17/29). However, the proportion was higher compared to those treated with PBS (0%, 0/31). Ep2 gene expression in the pituitary was > two-fold higher (P < 0.05) in the PGE2 group compared to the PBS and GnRH groups. Further, PGE2 stimulated Cox1 (2.7 fold, P < 0.05) while GnRH stimulated Cox2 expression (6.5 fold, P < 0.05) in the pituitary when compared to the PBS group. In conclusion, our results support the hypothesis that PGE2 can induce ovulation in prepubertal mice with a concomitant increase in Ep2 and Cox1 gene expression in the pituitary gland.