Objective: This study aims to explore mutation based on single nucleotide polymorphism (SNP) in the Cholecystokinin B receptor (CCKBR) gene of Pelung chickens. Materials and Methods: We collected DNA samples from 48 Pelung roosters that had won the crowing competition. The CCKBR target encompasses exon 3, intron 3, exon 4, and a part of intron 4, a long 601 bp. This target was replicated using PCR with specific primers that were designed by Primer-BLAST from NCBI. We generated the nucleotide sequence from the PCR product's sequencing results. The SNP analysis was done by BioEdit and MEGA. Genotyping and haplotyp¬ing were done based on nonsynonymous single nucleotide polymorphisms (SNPs) on exons 3 and 4. We calculated allele and genotype frequency, heterozygosity, and Hardy-Weinberg Equilibrium (HWE) using POPGENE 32 programs. Results: This study found three nonsynonymous single nucleotide polymorphisms. The nsSNP in exon 3 alters the coding for the 210th amino acid from serine to asparagine (g.1290 G > A/S210N), while the SNPs in exon 4 alter the coding for the 232nd amino acid from valine to phenylalanine (g.1423G > T/V232F) and the 243rd amino acid that changes the amino acid valine to glycine (g.1457T > G/V243G). The frequency of the mutated alleles is lower than the unmutated alleles. However, the mutation at position g.1457T > G/V243G produces a higher frequency than the unmutated allele. The allele and genotype frequency were not in HWE. It was caused by intensive selection in Pelung chickens, especially for growing capacity. Conclusion: Nonsynonymous mutation on CCKBR may cause variations in the crowing and other traits such as the growth of Pelung chickens. Further studies are needed to explore the CCKBR gene, including the relationship of the gene with the vigor and/or stress level of Pelung chickens. [J Adv Vet Anim Res 2025; 12(1.000): 141-148]

Objective: This study aims to explore mutation based on single nucleotide polymorphism (SNP) in the Cholecystokinin B receptor (CCKBR) gene of Pelung chickens. Materials and Methods: We collected DNA samples from 48 Pelung roosters that had won the crowing competition. The CCKBR target encompasses exon 3, intron 3, exon 4, and a part of intron 4, a long 601 bp. This target was replicated using PCR with specific primers that were designed by Primer-BLAST from NCBI. We generated the nucleotide sequence from the PCR product’s sequencing results. The SNP analysis was done by BioEdit and MEGA. Genotyping and haplotyping were done based on nonsynonymous single nucleotide polymorphisms (SNPs) on exons 3 and 4. We calculated allele and genotype frequency, heterozygosity, and Hardy-Weinberg Equilibrium (HWE) using POPGENE 32 programs. Results: This study found three nonsynonymous single nucleotide polymorphisms. The nsSNP in exon 3 altersthe coding for the 210th amino acid from serine to asparagine (g.1290 G > A/S210N), while the SNPs in exon 4 alter the coding for the 232nd amino acid from valine to phenylalanine (g.1423G > T/V232F) and the 243rd amino acid that changes the amino acid valine to glycine (g.1457T > G/V243G). The frequency of the mutated alleles is lower than the unmutated alleles. However, the mutation at position g.1457T > G/V243G produces a higher frequency than the unmutated allele. The allele and genotype frequency were not in HWE. It was caused by intensive selection in Pelung chickens, especially for growing capacity. Conclusion: Nonsynonymous mutation on CCKBR may cause variations in the crowing and other traits such as the growth of Pelung chickens. Further studies are needed to explore the CCKBR gene, including the relationship of the gene with the vigor and/or stress level of Pelung chickens. J. Adv. Vet. Anim. Res., 12(1): 141–148, March 2025 http://doi.org/10.5455/javar.2025.l881

Objectives: The rise of antibiotic-resistant strains of Staphylococcus aureus in dairy milk products is a global concern, compromising treatment efficacy and highlighting the need for innovative solutions. Therefore, a study was conducted to isolate S. aureus strains (N = 21) from raw milk samples of cows infected with subclinical bovine mastitis. Additionally, the resistance of these strains against 12 different antibiotics was examined. Materials and Methods: Sixty raw cow milk samples, 20 from each of three separate dairy farms in Lahore city, were collected and screened for the presence of S. aureus. It was discovered that 70% of these milk samples were contaminated with this bacterium, indicating a widespread presence across the farms. Different isolation tests were employed in this study, including gram staining, capsule staining, catalase, mannitol salt fermentation, DNase, coagulase, and oxidase. Results: The obtained results revealed that the isolated strains of S. aureus showed % of their resistance against different antibiotics in the order of amoxicillin (85%), penicillin (71%), gentamicin (CN) (42%), carbenicillin and trimethoprim-sulfamethoxazole (33%), streptomycin, ciprofloxacin, and oxytetracycline (28%), cefotaxime (10%), and chloramphenicol (4%) in decreasing order, respectively. However, these strains showed no resistance against Bacitracin and Ampicillin. Conclusion: The existence of resistant strains of S. aureus has been attributed to various factors, such as poor milk hygiene, delayed milk transportation, subclinical bovine mastitis among dairy cows, and antibiotic-resistant genes. Thus, our present study will provide useful information about the resistant strains of S. aureus, which may transfer through cows into milk and then produce serious food-borne diseases in human beings. This study will be helpful to improve and control the quality of dairy products in Pakistan. [J Adv Vet Anim Res 2025; 12(1.000): 252-259]

Objectives: The rise of antibiotic-resistant strains of Staphylococcus aureus in dairy milk products is a global concern, compromising treatment efficacy and highlighting the need for innovative solutions. Therefore, a study was conducted to isolate S. aureus strains (N = 21) from raw milk samples of cows infected with subclinical bovine mastitis. Additionally, the resistance of these strains against 12 different antibiotics was examined. Materials and Methods: Sixty raw cow milk samples, 20 from each of three separate dairy farms in Lahore city, were collected and screened for the presence of S. aureus. It was discovered that 70% of these milk samples were contaminated with this bacterium, indicating a widespread presence across the farms. Different isolation tests were employed in this study, including gram staining, capsule staining, catalase, mannitol salt fermentation, DNase, coagulase, and oxidase. Results: The obtained results revealed that the isolated strains of S. aureus showed % of their resistance against different antibiotics in the order of amoxicillin (85%), penicillin (71%), gentamicin (CN) (42%), carbenicillin and trimethoprim-sulfamethoxazole (33%), streptomycin, ciprofloxacin, and oxytetracycline (28%), cefotaxime (10%), and chloramphenicol (4%) in decreasing order, respectively. However, these strains showed no resistance against Bacitracin and Ampicillin. Conclusion: The existence of resistant strains of S. aureus has been attributed to various factors, such as poor milk hygiene, delayed milk transportation, subclinical bovine mastitis among dairy cows, and antibiotic-resistant genes. Thus, our present study will provide useful information about the resistant strains of S. aureus, which may transfer through cows into milk and then produce serious food-borne diseases in human beings. This study will be helpful to improve and control the quality of dairy products in Pakistan. J. Adv. Vet. Anim. Res., 12(1): 252–259, March 2025 http://doi.org/10.5455/javar.2025.l892

Objective: This study aimed to evaluate how well fresh broiler meat may be preserved in cold storage using chitosan–stearin as an edible coating. Materials and Methods: A completely randomized design with a 3 x 5 factorial arrangement and three replications was employed. Factor I represented the formula dosage (FD) (FD0 = 0% chitosan + 0% stearin; FD1 = 1.5% chitosan + 1% stearin; FD2 = 3% chitosan + 1% stearin), while Factor II represented storage time (ST) (ST0 = 0 days; ST1 = 3 days; ST2 = 6 days; ST3 = 9 days; ST4 = 12 days). Results: The results showed that the water content, cooking loss, protein content, and fat content of broiler meat were significantly affected (p < 0.01) by the FD and ST. Nonetheless, no significant difference (p > 0.05) was observed in the meat's ability to hold water. The broiler meat's pH was significantly affected (p < 0.01) by the FD, but it was not significantly affected (p > 0.05) by the ST. Furthermore, no treatment underwent testing, which revealed the presence of Escherichia coli and Salmonella sp. Conclusion: Chitosan–stearin edible coatings with different formula doses FD and ST consistently preserve the quality of fresh broiler meat during cold storage, with average values of water content ranging from 48.97% to 53.73%, water-holding capacity from 17.52% to 34.30%, cooking loss from 10.03% to 33.19%, pH levels from 4.93 to 5.53, protein content from 14.54% to 17.46%, fat content from 20.55% to 24.21%, and no detectable presence of E. coli and Salmonella sp. [J Adv Vet Anim Res 2025; 12(1.000): 99-105]

Objective: This study aimed to evaluate how well fresh broiler meat may be preserved in cold storage using chitosan–stearin as an edible coating. Materials and Methods: A completely randomized design with a 3 x 5 factorial arrangement and three replications was employed. Factor I represented the formula dosage (FD) (FD0 = 0% chitosan + 0% stearin; FD1 = 1.5% chitosan + 1% stearin; FD2 = 3% chitosan + 1% stearin), while Factor II represented storage time (ST) (ST0 = 0 days; ST1 = 3 days; ST2 = 6 days; ST3 = 9 days; ST4 = 12 days). Results: The results showed that the water content, cooking loss, protein content, and fat content of broiler meat were significantly affected (p < 0.01) by the FD and ST. Nonetheless, no significant difference (p > 0.05) was observed in the meat’s ability to hold water. The broiler meat’s pH was significantly affected (p < 0.01) by the FD, but it was not significantly affected (p > 0.05) by the ST. Furthermore, no treatment underwent testing, which revealed the presence of Escherichia coli and Salmonella sp. Conclusion: Chitosan–stearin edible coatings with different formula doses FD and ST consistently preserve the quality of fresh broiler meat during cold storage, with average values of water content ranging from 48.97% to 53.73%, water-holding capacity from 17.52% to 34.30%, cooking loss from 10.03% to 33.19%, pH levels from 4.93 to 5.53, protein content from 14.54% to 17.46%, fat content from 20.55% to 24.21%, and no detectable presence of E. coli and Salmonella sp. J. Adv. Vet. Anim. Res., 12(1): 99–105, March 2025 http://doi.org/10.5455/javar.2025.l876

Objective: This study aimed to compare two approaches to the brachial plexus: the traditional blind method and an additional technique, both of which use anatomical landmarks to guide needle insertion. Materials and Methods: The traditional and additional approaches were performed on both thoracic limbs of 24 cadavers (24 for each approach). Methylene dye is used for injection and nerve staining. We counted the nerves that successfully stained (staining less than 1 cm). The Mann–Whitney U test was used to compare approaches. Results: The findings indicated that all cadavers were successfully used. The traditional approach and the additional approach revealed success rates of 45.83% and 54.17%, respectively. We found no significant differences between the two techniques (p > 0.05). Conclusion: The additional approach presents a viable alternative to the traditional method for performing the brachial plexus block in goats. Further research into the clinical differences between these techniques could lead to useful insights that help make them more accurate and useful. [J Adv Vet Anim Res 2025; 12(1.000): 64-69]

Objective: This study aimed to compare two approaches to the brachial plexus: the traditional blind method and an additional technique, both of which use anatomical landmarks to guide needle insertion. Materials and Methods: The traditional and additional approaches were performed on both thoracic limbs of 24 cadavers (24 for each approach). Methylene dye is used for injection and nerve staining. We counted the nerves that successfully stained (staining less than 1 cm). The Mann– Whitney U test was used to compare approaches. Results: The findings indicated that all cadavers were successfully used. The traditional approach and the additional approach revealed success rates of 45.83% and 54.17%, respectively. We found no significant differences between the two techniques (p > 0.05). Conclusion: The additional approach presents a viable alternative to the traditional method for performing the brachial plexus block in goats. Further research into the clinical differences between these techniques could lead to useful insights that help make them more accurate and useful. J. Adv. Vet. Anim. Res., 12(1): 64–69, March 2025 http://doi.org/10.5455/javar.2025.l872

Objective: This study aimed to assess Aerodramus fuciphagus (Thunberg, 1812) nest's mineral content, cytotoxicity effects, and antioxidant activity. The Edible birds' nests (EBNs) were obtained from Tanah Merah (TM), Tumpat (TU), and Jeli (JE) districts in Kelantan, Malaysia. Materials and Methods: The mineral content was assessed via inductively coupled plasma mass spectrometry. The cytotoxicity effect was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide and brine shrimp lethality assays. The antioxidant activity was investigated by 2,2´-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and ferric-reducing antioxidant power (FRAP) assays. Results: Valuable macro- and microelements, including sodium, potassium, aluminum, calcium, magnesium, zinc, iron, and selenium, were detected in the EBN samples. The cytotoxicity test revealed that JE hydrochloric acid (HCl) extract had promising efficacy against HeLa cell lines, with IC50 values of 20.00 ± 0.57 μg/ml. Similarly, TU sodium hydroxide (NaOH) extract inhibited the proliferation of MCF-7 cell lines (IC50 0.03 ± 0.01 μg/ml). Based on the BSL assay, the EBNs were considered safe for consumption as the LC50 values of all the samples were below the toxic level (>1,000 μg/ml). TM EBNs aqueous, TU aqueous, and HCl extracts showed promising antioxidant activity at IC50 values (10 μg/ml) in the DPPH assay. However, the NaOH extract from TU exhib¬ited the highest FRAP value (1.38 ± 0.00 mM gallic acid equivalents gm⁻¹) compared to all other extracts. Conclusion: This study's findings demonstrated the potential of EBN as an alternative anticancer agent and natural antioxidant. [J Adv Vet Anim Res 2025; 12(1.000): 132-140]

Objective: This study aimed to assess Aerodramus fuciphagus (Thunberg, 1812) nest’s mineral content, cytotoxicity effects, and antioxidant activity. The Edible birds’ nests (EBNs) were obtained from Tanah Merah (TM), Tumpat (TU), and Jeli (JE) districts in Kelantan, Malaysia. Materials and Methods: The mineral content was assessed via inductively coupled plasma mass spectrometry. The cytotoxicity effect was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide and brine shrimp lethality assays. The antioxidant activity was investigated by 2,2´-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and ferric-reducing antioxidant power (FRAP) assays. Results: Valuable macro- and microelements, including sodium, potassium, aluminum, calcium, magnesium, zinc, iron, and selenium, were detected in the EBN samples. The cytotoxicity test revealed that JE hydrochloric acid (HCl) extract had promising efficacy against HeLa cell lines, with IC50 values of 20.00 ± 0.57 µg/ml. Similarly, TU sodium hydroxide (NaOH) extract inhibited the proliferation of MCF-7 cell lines (IC50 0.03 ± 0.01 µg/ml). Based on the BSL assay, the EBNs were considered safe for consumption as the LC50 values of all the samples were below the toxic level (>1,000 µg/ml). TM EBNs aqueous, TU aqueous, and HCl extracts showed promising antioxidant activity at IC50 values (10 µg/ml) in the DPPH assay. However, the NaOH extract from TU exhibited the highest FRAP value (1.38 ± 0.00 mM gallic acid equivalents gm⁻¹) compared to all other extracts. Conclusion: This study’s findings demonstrated the potential of EBN as an alternative anticancer agent and natural antioxidant. J. Adv. Vet. Anim. Res., 12(1): 132–140, March 2025 http://doi.org/10.5455/javar.2025.l880