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: A phylogenetic study was carried out on the avian influenza virus (AIV) isolated from a disease outbreak in Sidenreng Rappang Regency, South Sulawesi, Indonesia, in 2018.  Material and Methods: Oropharyngeal swabs and organ samples were obtained from ducks that showed clinical symptoms: torticollis, fascial edema, neurological disorders, the corneas appear cloudy, and death occurs less than 1 day after symptoms appear. In this study, isolate A/duck/ Sidenreng Rappang/07180110-11/2018 from duck was sequenced and characterized.  Results: It was found that each gene segment of the virus has the highest nucleotide homology to the Indonesian highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.2.1c. Multiple alignments of the sample Hemagglutinin (HA) gene with the avian influenza references virus showed that the pattern of amino acid arrangement in the cleavage site PQRERRRK-RGLF is the characteristic of the HPAI virus. In addition, the HA gene contained Q222 (glutamine) and G224 (glycine), signify­ing a high affinity to avian receptor binding specificity (SA α2,3 Gal). Furthermore, there was no genetic reassortment of this virus based on the phylogenetic analysis of HA, NA, PB1, PB2, PA, NP, M, and NS genes.  Conclusion: The HPAI H5N1 clade 2.3.2.1c virus was identified in duck farms in South Sulawesi, Indonesia. J. Adv. Vet. Anim. Res., 8(2): 346-354, June 2021

Objective: The avian influenza virus (AIV) subtype H9N2 circulating in Indonesia has raised increasing concern about its impact on poultry and its public health risks. In this study, the H9N2 virus from chicken poultry farms in Java was isolated and characterized molecularly.  Materials and Methods: Thirty-three pooled samples of chicken brain, cloacal swab, trachea, and oviduct were taken from multiple chickens infected with AIV in five regions of Java, Indonesia. The samples were isolated from specific pathogenic-free embryonated eggs that were 9 days old. Reverse transcription polymerase chain reaction and sequencing were used to identify H9N2 viruses.  Results: This study was successful in detecting and characterizing 13 H9N2 isolates. The sequenc­ing analysis of hemagglutinin genes revealed a 96.9%–98.8% similarity to the H9N2 AIV isolated from Vietnam in 2014 (A/muscovy duck/Vietnam/LBM719/2014). According to the phylogenetic analysis, all recent H9N2 viruses were members of the lineage Y280 and clade h9.4.2.5. Nine of the H9N2 isolates studied showed PSKSSR↓GLF motifs at the cleavage site, while four had PSKSSR↓GLF. Notably, all contemporary viruses have leucine (L) at position 216 in the recep­tor-binding region, indicating that the virus can interact with a human-like receptor.  Conclusion: This study described the features of recent H9N2 viruses spreading in Java’s poul­try industry. Additionally, H9N2 infection prevention and management must be implemented to avoid the occurrence of virus mutations in the Indonesian poultry industry. J. Adv. Vet. Anim. Res., 8(2): 423-434, June 2021 http://doi.org/10.5455/javar.2021.h530

Objective: The purpose of this study was to compare the effects of pretreatment with medetomidine (Me), midazolam (Mi), and ketamine (Ke) on stress-related neurohormonal and metabolic responses in isoflurane-anesthetized cats undergoing ovariohysterectomy and castration. Materials and Methods: We prospectively recruited 112 client-owned healthy mixed-breed cats. In both surgeries, we divided the cats into seven groups (eight cats per group): non-treatment (control), Me (50 μg/kg), Mi (0.5 mg/kg), Ke (5 mg/kg), Me + Mi, Me + Ke and Me + Mi + Ke administered intramuscularly. After pretreatments, we maintained anesthesia with isoflurane and oxygen. Venous blood was taken before pretreatment, pre- and post-operatively during anesthesia, and at early- and complete-recovery. Results: Both plasma adrenaline and noradrenaline were reduced during anesthesia in all groups. Plasma cortisol increased during anesthesia and at early recovery in non-Me-treated groups, whereas it decreased in Me-treated groups in both surgeries. Plasma insulin and non-esterified fatty acid (NEFA) decreased, and glucose increased during anesthesia in all groups, but hyperglycemia and decrease in NEFA were greater in Me-treated groups. Conclusions: In isoflurane-anesthetized cats undergoing surgeries, premedication with Me alone and in combination is useful for reducing the perioperative stress-related increase in cortisol and catecholamines except for hyperglycemia. J. Adv. Vet. Anim. Res., 8(4): 563-575, December 2021 http://doi.org/10.5455/javar.2021.h546

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). PPE’s 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 meat’s freshness while being stored at a refrigerator temperature (5°C ± 1°C). J. Adv. Vet. Anim. Res., 8(4): 612–618, December 2021 http://doi.org/10.5455/javar.2021.h552

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 viabil­ity and progressive motility up to 96 h independently of the extender used. J. Adv. Vet. Anim. Res., 8(4): 635–641, December 2021 http://doi.org/10.5455/javar.2021.h555

Objective: In developing countries, such as Bangladesh, the live bird market (LBM) is a vital location for the trading of live poultry. The study was carried out in nine LBMs located around Bangladesh to ascertain the present regulations and procedures governing their operation. Additionally, the responsibilities and levels of engagement of the stakeholders were determined.  Materials and Methods: The data were gathered through the use of a semi-structured interview guide. Thematic analysis was used to code the interview transcripts iteratively.  Results: The findings indicated that the government was directly and indirectly involved in the leasing process of the markets. A market in this country is divided into numerous sectors, includ­ing LBM, fish market, vegetable market, and grocery stores. A market’s hygienic condition is highly dependent on market authority’s decisions. In some markets, market officials conducted routine sanitary inspections. Veterinarians played a little role in the inspection procedure.  Conclusion: There is no adequate, functional monitoring system to ensure that LBMs adhere to cleanliness and adequate and functional biosecurity. Biosecurity enhancements, effective clean­ing programs, and regular monitoring by relevant authorities are critical for LBMs in Bangladesh. J. Adv. Vet. Anim. Res., 8(4): 671–678, December 2021 http://doi.org/10.5455/javar.2021.h559