Objective: This short study describes the occurrence of pathogenic Leptospira spp. in two major wet markets in Kota Bharu, Kelantan, Malaysia. Materials and Methods: 30 rodents (20 rats and 10 shrews) were caught in 2 wet markets, and a postmortem was performed to extract both kidneys. Molecular diagnosis via polymerase chain reaction (PCR) was conducted to detect leptospiral DNA using universal and pathogenic Leptospira primers, respectively. Results: The results showed that 20/28 (72%) rat samples were detected positive for Leptospira spp, and all shrews were negative. Further sequencing analysis identified L. interrogans and L. borgpetersenii as the most frequently Leptospirosis species from kidney samples. Conclusions: The presented study here sheds light on the presence of pathogenic leptospires harboring the rat population in both wet markets in Kelantan, which presents a great public health risk to wet market workers and visitors. J. Adv. Vet. Anim. Res., 9(2): 275–281, June 2022 http://doi.org/10.5455/javar.2022.i593

Objective: This short study describes the occurrence of pathogenic Leptospira spp. in two major wet markets in Kota Bharu, Kelantan, Malaysia. Materials and Methods: 30 rodents (20 rats and 10 shrews) were caught in 2 wet markets, and a postmortem was performed to extract both kidneys. Molecular diagnosis via polymerase chain reaction (PCR) was conducted to detect leptospiral DNA using universal and pathogenic Leptospira primers, respectively. Results: The results showed that 20/28 (72%) rat samples were detected positive for Leptospira spp, and all shrews were negative. Further sequencing analysis identified L. interrogans and L. borgpetersenii as the most frequently Leptospirosis species from kidney samples. Conclusions: The presented study here sheds light on the presence of pathogenic leptospires har¬boring the rat population in both wet markets in Kelantan, which presents a great public health risk to wet market workers and visitors. [J Adv Vet Anim Res 2022; 9(2.000): 275-281]

Objective: This short study describes the occurrence of pathogenic Leptospira spp. in two major wet markets in Kota Bharu, Kelantan, Malaysia. Materials and Methods: 30 rodents (20 rats and 10 shrews) were caught in 2 wet markets, and a postmortem was performed to extract both kidneys. Molecular diagnosis via polymerase chain reaction (PCR) was conducted to detect leptospiral DNA using universal and pathogenic Leptospira primers, respectively. Results: The results showed that 20/28 (72%) rat samples were detected positive for Leptospira spp, and all shrews were negative. Further sequencing analysis identified L. interrogans and L. borgpetersenii as the most frequently Leptospirosis species from kidney samples. Conclusions: The presented study here sheds light on the presence of pathogenic leptospires har¬boring the rat population in both wet markets in Kelantan, which presents a great public health risk to wet market workers and visitors. [J Adv Vet Anim Res 2022; 9(2.000): 275-281]

Objective: As one of the most valuable genetic resources of Ongole beef cattle globally, the Sumba Ongole (SO) cattle population is being studied in this investigation of myostatin (MSTN) gene polymorphism and its association with growth traits. Materials and Methods: Blood samples from 161 SO cattle were collected and analyzed. Deoxyribonucleic acid (DNA) was isolated. The DNA was electrophoresed and extracted, and finally, the annealing temperature was optimized, followed by amplification and sequencing. Next, we used a Basic local alignment search tool to assess the sequencing data. Results: The analysis revealed 22 single nucleotide polymorphisms (SNPs) in the MSTN gene in this region that showed genetic variation. Two SNPs, c.424 G > A, and c.467 G > C, were found to be significantly associated with SO cattle phenotypes of wither height, heart girth, and hip height (p < 0.05) but not with body weight or body length (p > 0.05). Conclusion: As a result of our findings, the MSTN gene polymorphism and its correlation with growth traits in SO cattle may be employed as a candidate marker in SO cattle and other beef cattle breeds. J. Adv. Vet. Anim. Res., 9(4): 565–572, December 2022 http://doi.org/10.5455/javar.2022.i625

Objective: As one of the most valuable genetic resources of Ongole beef cattle globally, the Sumba Ongole (SO) cattle population is being studied in this investigation of myostatin (MSTN) gene polymorphism and its association with growth traits. Materials and Methods: Blood samples from 161 SO cattle were collected and analyzed. Deoxyribonucleic acid (DNA) was isolated. The DNA was electrophoresed and extracted, and finally, the annealing temperature was optimized, followed by amplification and sequencing. Next, we used a Basic local alignment search tool to assess the sequencing data. Results: The analysis revealed 22 single nucleotide polymorphisms (SNPs) in the MSTN gene in this region that showed genetic variation. Two SNPs, c.424 G > A, and c.467 G > C, were found to be significantly associated with SO cattle phenotypes of wither height, heart girth, and hip height (p < 0.05) but not with body weight or body length (p > 0.05). Conclusion: As a result of our findings, the MSTN gene polymorphism and its correlation with growth traits in SO cattle may be employed as a candidate marker in SO cattle and other beef cattle breeds. [J Adv Vet Anim Res 2022; 9(4.000): 565-572]

Objective: This study describes the newest deoxyribonucleic acid (DNA) barcoding Asian redtail catfish (Hemibagrus nemurus) in the Progo River, Magelang, Central Java, Indonesia. Materials and Methods: Ten fish were caught in the Progo River, Magelang, Central Java, Indonesia. The polymerase chain reaction was the molecular diagnosis to detect the sequences of DNA of Cytochrome Oxidase I compared to National Center for Biotechnology Information data (GenBank). Results: The results showed that the percent identity was not 100% with H. nemurus data from other locations (GenBank), including Indonesia. The closest percent identity was H. nemurus from Java Island (Accession ID: MK312566.1) with 97.6% similarity. The genetic mutation that happened might be due to environmental change (pollution) in the Progo River recently. Conclusions: This study showed a genetic mutation in H. nemurus from Progo River may be caused by environmental change. Low pollution exposure levels may not be detrimental (lethal) to fish. However, it can affect fish fertility, which leads to population degradation (gene variation). Attention must be increased for fish survival in the future. J. Adv. Vet. Anim. Res., 9(4): 591–600, December 2022 http://doi.org/10.5455/javar.2022.i628

Objective: This study describes the newest deoxyribonucleic acid (DNA) barcoding Asian redtail catfish (Hemibagrus nemurus) in the Progo River, Magelang, Central Java, Indonesia. Materials and Methods: Ten fish were caught in the Progo River, Magelang, Central Java, Indonesia. The polymerase chain reaction was the molecular diagnosis to detect the sequences of DNA of Cytochrome Oxidase I compared to National Center for Biotechnology Information data (GenBank). Results: The results showed that the percent identity was not 100% with H. nemurus data from other locations (GenBank), including Indonesia. The closest percent identity was H. nemurus from Java Island (Accession ID: MK312566.1) with 97.6% similarity. The genetic mutation that hap¬pened might be due to environmental change (pollution) in the Progo River recently. Conclusions: This study showed a genetic mutation in H. nemurus from Progo River may be caused by environmental change. Low pollution exposure levels may not be detrimental (lethal) to fish. However, it can affect fish fertility, which leads to population degradation (gene variation). Attention must be increased for fish survival in the future. [J Adv Vet Anim Res 2022; 9(4.000): 591-600]

Objective: This study was conducted to determine the colony forming units (CFU) to isolate, identify, and antibiotic sensitivity of Staphylococcus aureus from chicken nuggets (CN). Materials and Methods: Sixty CN samples from two brands were collected from different superstores in Mymensingh, Bangladesh. Uncooked, oven-cooked (OC), or gas stove-cooked (GSC) CN samples were inoculated onto mannitol salt agar and blood agar. Results: The total staphylococcal count (TSC) for uncooked CN ranged from log 4.68 to log 5.11 CFU/gm. For OC CN, TSC ranged from log 3.29 to log 3.62 CFU/gm. For GSC CN, TSC ranged from log 3.09 to log 3.49 CFU/gm. Relative to uncooked CN, microwave oven-cooked and GSC samples significantly reduced the TSC of CN(p < 0.01). Using the polymerase chain reaction assay and standard biochemical testing, only 8 out of 60 CN samples contained S. aureus. Staphylococcus aureus were resistant to Ampicillin (100%), Amoxicillin (100%), Oxacillin (75%), Cefixime (87.5%), Doxycycline (75%), intermediately sensitive to Erythromycin (25%), Cephalexin (12.5%), Ciprofloxacin (25%), Gentamicin (12.5%), Doxycycline (12.5%) and sensitive to Oxacillin (25%), Azithromycin (100%), Erythromycin (75%) Cephalexin (87.5%), Cefixime (12.5%), Chloramphenicol (100%), Ciprofloxacin (75%), Gentamicin (87.5%), Doxycycline (12.5%), and Vancomycin (100%). Conclusion: This study reports the first isolation and identification of S. aureus from CN in Bangladesh. GSC CN was better than OC and uncooked CN. Data also suggest that CN is contaminated with multidrug-resistant S. aureus, which poses a public health hazard. J. Adv. Vet. Anim. Res., 9(4): 601–609, December 2022 http://doi.org/10.5455/javar.2022.i629

Objective: This study was conducted to determine the colony forming units (CFU) to isolate, identify, and antibiotic sensitivity of Staphylococcus aureus from chicken nuggets (CN). Materials and Methods: Sixty CN samples from two brands were collected from different super¬stores in Mymensingh, Bangladesh. Uncooked, oven-cooked (OC), or gas stove-cooked (GSC) CN samples were inoculated onto mannitol salt agar and blood agar. Results: The total staphylococcal count (TSC) for uncooked CN ranged from log 4.68 to log 5.11 CFU/gm. For OC CN, TSC ranged from log 3.29 to log 3.62 CFU/gm. For GSC CN, TSC ranged from log 3.09 to log 3.49 CFU/gm. Relative to uncooked CN, microwave oven-cooked and GSC samples significantly reduced the TSC of CN (p < 0.01). Using the polymerase chain reaction assay and standard biochemical testing, only 8 out of 60 CN samples contained S. aureus. Staphylococcus aureus were resistant to Ampicillin (100%), Amoxicillin (100%), Oxacillin (75%), Cefixime (87.5%), Doxycycline (75%), intermediately sensitive to Erythromycin (25%), Cephalexin (12.5%), Ciprofloxacin (25%), Gentamicin (12.5%), Doxycycline (12.5%) and sensitive to Oxacillin (25%), Azithromycin (100%), Erythromycin (75%) Cephalexin (87.5%), Cefixime (12.5%), Chloramphenicol (100%), Ciprofloxacin (75%), Gentamicin (87.5%), Doxycycline (12.5%), and Vancomycin (100%). Conclusion: This study reports the first isolation and identification of S. aureus from CN in Bangladesh. GSC CN was better than OC and uncooked CN. Data also suggest that CN is contami¬nated with multidrug-resistant S. aureus, which poses a public health hazard. [J Adv Vet Anim Res 2022; 9(4.000): 601-609]

Objective: The study was conducted to determine the marbling score, fat and meat color, cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and physical–chemical content of male Bali beef fed fermented pineapple peel. Materials and Methods: Twelve heads of male Bali cattle with an initial weight of 168.46 ± 11.95 kg were put into individual cages at random based on a completely randomized design with three treatments and four heads of Bali cattle as replicates, namely T0 = NG + (39% MC + 61% RB + 0% fermented pineapple peel); T1 = NG + (10% MC + 70% RB + 20% fermented pineapple peel with yeast culture); and T2 = NG + (15% MG + 65% RB + 20% pineapple peel fermented by lactic acid bacteria). The sample of Bali cattle meat used in testing the research variables was the LD muscle, with as many as 24 samples for each treatment. The data were analyzed based on the analysis of variance using the Statistical Product and Service Solutions software program, following Duncan’s test with 5% confidence. Results: The results showed that the treatment T2 could increase the marbling quality of the meat from 2.58% to 4.00%. The cholesterol content (80 mg/100 gm), HDL (60 mg/100 gm), LDL (10 mg/100 gm), water-holding capacity (36.10%), cooking loss (29.16%), tenderness/shear force (4.08 kg/cm2), crude protein (22.99%), crude fat (4.23%), and meat collagen (1.65%) were determined. Conclusions: Adding 20% of fermented pineapple peel by lactic acid bacteria to the ration can improve the quality of marbling, produce cholesterol, and the physical–chemical value of meat that meets the Indonesian National Standard. J. Adv. Vet. Anim. Res., 9(3): 419–431, September 2022 http://doi.org/10.5455/javar.2022.i610

Objective: The study was conducted to determine the marbling score, fat and meat color, choles¬terol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and physical–chemical con¬tent of male Bali beef fed fermented pineapple peel. Materials and Methods: Twelve heads of male Bali cattle with an initial weight of 168.46 ± 11.95 kg were put into individual cages at random based on a completely randomized design with three treatments and four heads of Bali cattle as replicates, namely T0 = NG + (39% MC + 61% RB + 0% fermented pineapple peel); T1 = NG + (10% MC + 70% RB + 20% fermented pineapple peel with yeast culture); and T2 = NG + (15% MG + 65% RB + 20% pineapple peel fermented by lactic acid bacteria). The sample of Bali cattle meat used in testing the research variables was the LD muscle, with as many as 24 samples for each treatment. The data were analyzed based on the analysis of variance using the Statistical Product and Service Solutions software program, following Duncan's test with 5% confidence. Results: The results showed that the treatment T2 could increase the marbling quality of the meat from 2.58% to 4.00%. The cholesterol content (80 mg/100 gm), HDL (60 mg/100 gm), LDL (10 mg/100 gm), water-holding capacity (36.10%), cooking loss (29.16%), tenderness/shear force (4.08 kg/cm2), crude protein (22.99%), crude fat (4.23%), and meat collagen (1.65%) were determined. Conclusions: Adding 20% of fermented pineapple peel by lactic acid bacteria to the ration can improve the quality of marbling, produce cholesterol, and the physical–chemical value of meat that meets the Indonesian National Standard. [J Adv Vet Anim Res 2022; 9(3.000): 419-431]

Objective: To elucidate the cellular mechanisms of polyherbal formulation [Kolin PlusTM (KP)], genomics was performed to delineate the genes and pathways associated with lipid regulation through transcriptional profiling of the liver in commercial broilers raised on diets deficient in choline chloride (CCL). Materials and Methods: The gene expression patterns were studied for four groups [normal diet: normal, choline chloride deficient (CCD), KP (400 gm/ton), and CCL (400 gm/ton)] using Agilent microarray on day 42. The hierarchical cluster analysis was carried out on 12,614 differentially expressed genes (DEGs) with a similar expression. Results: Out of 12,614 significant DEGs, 1,926, 448, and 1,330 genes were expressed at higher rates, and 413, 482, and 1,364 were expressed at lower rates than CCD (CCD vs. normal), CCL (CCL vs. CCD), and KP (KP vs. CCD), respectively. GO enrichment analysis of DEG further revealed the significant association of biological process items with the lipid, sterol, and lipoprotein metabolic processes. In particular, peroxisome proliferator-activated receptor gamma coactivator 1 alpha, carnitine palmitoyl transferase I, hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit beta, and patatin-like phospholipase domain containing 2 genes involved in fatty acid oxidation and lipase C, ABCG5, ABCG8, acetyl-CoA carboxylase, ATP citrate lyase enzyme, and peroxisome proliferator-activated receptor gamma genes involved in lipogenesis were altered by KP intervention for lipid metabolism. Conclusions: These findings reveal that the supplementation of KP prevents fatty liver-associated problems in broiler chickens by modulating the expression of the above-mentioned genes that are responsible for the oxidation of fatty acids and lipogenesis in the liver. J. Adv. Vet. Anim. Res., 9(3): 432–439, September 2022 http://doi.org/10.5455/javar.2022.i611

Objective: To elucidate the cellular mechanisms of polyherbal formulation [Kolin PlusTM (KP)], genomics was performed to delineate the genes and pathways associated with lipid regulation through transcriptional profiling of the liver in commercial broilers raised on diets deficient in choline chloride (CCL). Materials and Methods: The gene expression patterns were studied for four groups [normal diet: normal, choline chloride deficient (CCD), KP (400 gm/ton), and CCL (400 gm/ton)] using Agilent microarray on day 42. The hierarchical cluster analysis was carried out on 12,614 differentially expressed genes (DEGs) with a similar expression. Results: Out of 12,614 significant DEGs, 1,926, 448, and 1,330 genes were expressed at higher rates, and 413, 482, and 1,364 were expressed at lower rates than CCD (CCD vs. normal), CCL (CCL vs. CCD), and KP (KP vs. CCD), respectively. GO enrichment analysis of DEG further revealed the significant association of biological process items with the lipid, sterol, and lipoprotein metabolic processes. In particular, peroxisome proliferator-activated receptor gamma coactivator 1 alpha, carnitine palmitoyl transferase I, hydroxyacyl-CoA dehydrogenase trifunctional multienzyme com¬plex subunit beta, and patatin-like phospholipase domain containing 2 genes involved in fatty acid oxidation and lipase C, ABCG5, ABCG8, acetyl-CoA carboxylase, ATP citrate lyase enzyme, and peroxisome proliferator-activated receptor gamma genes involved in lipogenesis were altered by KP intervention for lipid metabolism. Conclusions: These findings reveal that the supplementation of KP prevents fatty liver-associated problems in broiler chickens by modulating the expression of the above-mentioned genes that are responsible for the oxidation of fatty acids and lipogenesis in the liver. [J Adv Vet Anim Res 2022; 9(3.000): 432-439]

Objective: The increase and prevalence of multidrug-resistant bacteria in livestock animals are serious public health concerns. This study aimed to identify the presence of the blaTEM gene in extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolated from rectal swabs of apparently healthy pigs in Malang District, East Java, Indonesia. Materials and Methods: A total of 120 rectal swab samples were collected from the pigs. The rectal swabs were screened for the presence of E. coli using standard microbiological identification procedures. The Kirby–Bauer disk diffusion method identified multidrug-resistant E. coli. Five different classes of antibiotics were used to identify multidrug-resistant isolates, including Ciprofloxacin, Trimethoprim, Tetracycline, Streptomycin, and Aztreonam. Multidrug-resistant E. coli isolates were characterized for the presence of ESBL using double-disk synergy test methods. The presence of blaTEM genes was determined using polymerase chain reaction methods. Results: The results of this study indicated that 107 (89.2%) out of 120 samples analyzed were positive for E. coli isolates. A total of 32 (29.9%) E. coli isolates were identified to be multidrug-resistant and further subjected to molecular testing. The molecular analysis revealed (5; 15.6%) E. coli isolates to harbor the blaTEM gene. Conclusion: The results of this study revealed that pigs and products of pork origin must be considered a source of transmission of ESBL-producing E. coli to public health important under the food chain. J. Adv. Vet. Anim. Res., 9(3): 447–452, September 2022 http://doi.org/10.5455/javar.2022.i613

Objective: The increase and prevalence of multidrug-resistant bacteria in livestock animals are serious public health concerns. This study aimed to identify the presence of the blaTEM gene in extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolated from rectal swabs of apparently healthy pigs in Malang District, East Java, Indonesia. Materials and Methods: A total of 120 rectal swab samples were collected from the pigs. The rectal swabs were screened for the presence of E. coli using standard microbiological identifi¬cation procedures. The Kirby–Bauer disk diffusion method identified multidrug-resistant E. coli. Five different classes of antibiotics were used to identify multidrug-resistant isolates, including Ciprofloxacin, Trimethoprim, Tetracycline, Streptomycin, and Aztreonam. Multidrug-resistant E. coli isolates were characterized for the presence of ESBL using double-disk synergy test methods. The presence of blaTEM genes was determined using polymerase chain reaction methods. Results: The results of this study indicated that 107 (89.2%) out of 120 samples analyzed were positive for E. coli isolates. A total of 32 (29.9%) E. coli isolates were identified to be multidrug-re¬sistant and further subjected to molecular testing. The molecular analysis revealed (5; 15.6%) E. coli isolates to harbor the blaTEM gene. Conclusion: The results of this study revealed that pigs and products of pork origin must be considered a source of transmission of ESBL-producing E. coli to public health important under the food chain. [J Adv Vet Anim Res 2022; 9(3.000): 447-452]

Objectives: This research aimed to isolate, identify, and characterize a new strain of Bacillus cereus through different molecular biology approaches so that it could be further studied for therapeutic purposes against selective enteric pathogens. Materials and Methods: Pure isolates of B. cereus were prepared from buffalo yogurt samples in REMBA medium. Initially, the morphological, physiological, and biochemical properties were studied accordingly. Following the tests, the molecular identification for the strain identification was conducted through plasmid DNA extraction, PCR, agarose gel electrophoresis, and 16S rRNA sequencing up to 1.37 kb. Afterward, the antibiotic sensitivity [Epsilometer test (E-Test)] and antifungal activity were tested considering different concentrations. Being classified from the aforementioned tests, a comprehensive antimicrobial activity test was conducted using the cell-free-supernatant (CFS) of the test strain against selective enteric pathogens in humans in vitro. Besides, the different clusters of genes were identified and characterized for understanding the presumptive bacteriocins present in the CFS of the strain in silico, where molecular string properties were calculated. Finally, the evolutionary relationship among diversified bacteriocins synthesized by different Bacillus strains was studied to predict the CFS-containing bacteriocins of the new strain. Results: Purified isolates of B. cereus were Gram-positive rods and showed significant tolerance (p < 0.0001) to different concentrations of pH, phenol, bile salt, and NaCl. 16S rRNA revealed the strain as LOCK 1002, which was strongly sensitive to all the antibiotics used and resistant to selective antifungal agents. The CFS of B. cereus LOCK 1002 was found to be a very promising antagonist to all the enteric pathogens used in the culture condition. Two gene clusters were predicted to be interconnected and responsible for different presumptive bacteriocins. Conclusion: The newly identified LOCK 1002 can be a very potent strain of B. cereus in use as an antimicrobial agent for having different bacteriocin coding gene clusters. J. Adv. Vet. Anim. Res., 9(4): 663–675, December 2022 http://doi.org/10.5455/javar.2022.i635

Objectives: This research aimed to isolate, identify, and characterize a new strain of Bacillus cereus through different molecular biology approaches so that it could be further studied for therapeutic purposes against selective enteric pathogens. Materials and Methods: Pure isolates of B. cereus were prepared from buffalo yogurt samples in REMBA medium. Initially, the morphological, physiological, and biochemical properties were studied accordingly. Following the tests, the molecular identification for the strain identification was conducted through plasmid DNA extraction, PCR, agarose gel electrophoresis, and 16S rRNA sequencing up to 1.37 kb. Afterward, the antibiotic sensitivity [Epsilometer test (E-Test)] and anti¬fungal activity were tested considering different concentrations. Being classified from the aforementioned tests, a comprehensive antimicrobial activity test was conducted using the cell-free-su¬pernatant (CFS) of the test strain against selective enteric pathogens in humans in vitro. Besides, the different clusters of genes were identified and characterized for understanding the presump¬tive bacteriocins present in the CFS of the strain in silico, where molecular string properties were calculated. Finally, the evolutionary relationship among diversified bacteriocins synthesized by different Bacillus strains was studied to predict the CFS-containing bacteriocins of the new strain. Results: Purified isolates of B. cereus were Gram-positive rods and showed significant tolerance (p < 0.0001) to different concentrations of pH, phenol, bile salt, and NaCl. 16S rRNA revealed the strain as LOCK 1002, which was strongly sensitive to all the antibiotics used and resistant to selec¬tive antifungal agents. The CFS of B. cereus LOCK 1002 was found to be a very promising antago¬nist to all the enteric pathogens used in the culture condition. Two gene clusters were predicted to be interconnected and responsible for different presumptive bacteriocins. Conclusion: The newly identified LOCK 1002 can be a very potent strain of B. cereus in use as an antimicrobial agent for having different bacteriocin coding gene clusters. [J Adv Vet Anim Res 2022; 9(4.000): 663-675]