Objective: The percentage of infertility cases in this world is about 50%. Seahorses (Hippocampus spp.) are widely used in traditional medicine. Several studies suggest that seahorses have ethno pharmacological characteristics, such as fertility, antioxidants, and antifatigue. The purpose of this study was to determine whether seahorse extract (SE) (Hippocampus comes L.) has an effect on fertility and serum biochemistry in rats induced by depo medroxyprogesterone acetate (DMPA). Materials and Methods: All animals were induced with 1.25 mg/kg BW of DMPA. Animals were grouped into five groups: namely aquadest, 1% CMC, and SE doses of 150, 225, and 300 mg/kg BW. The rats were gavage every morning from week 7 until 18. At the end of our study, semen from the vas deferens and blood from the heart were analyzed. We analyzed with a one-way analysis of variance and Bonferroni’s post hoc tests (α 95%). Results: The concentration of spermatozoa had a significant difference in dose of 150 mg/kg BW compared to other groups (p = 0.04). In contrast, the motility (p = 0.012) and viability of spermatozoa (p = 0.007) were highly significant differences (p < 0.05 and p < 0.01) at 300 mg/kg BW. Testosterone levels were not significantly (p = 0.162; p > 0.05), but tended to increase at 300 mg/ kg BW (11.01%). Nevertheless, serum biochemistry was insignificant (p > 0.05) in all groups. Conclusion: SE (Hippocampus comes L.) ameliorates fertility and serum biochemistry in rats induced by DMPA. J. Adv. Vet. Anim. Res., 10(1): 126–131, March 2023 http://doi.org/10.5455/javar.2023.j661

Objective: The percentage of infertility cases in this world is about 50%. The seahorse (Hippocampus spp.) is wide to traditional medicine. Several studies stated that seahorse has eth¬nopharmacological features, such as fertility, antioxidants, and antifatigue. The purpose of this study was to determine whether seahorse extract (SE) (Hippocampus comes L.) affects fertility and serum biochemistry in rats induced by depo medroxyprogesterone acetate (DMPA). Materials and Methods: All animals were induced by 1.25 mg/kg BW DMPA. The animals were grouped into five groups: aquadest, CMC 1%, and SE doses of 150, 225, and 300 mg/kg BW. The rats were gavage every morning from week 7 until 18. At the end of our study, the semen from the vas deferens and blood from the heart were analyzed. We analyzed with a one-way analysis of variance and Bonferroni's post hoc tests (α 95%). Results: The concentration of spermatozoa had a significant difference in dose of 150 mg/kg BW compared to other groups (p = 0.04). In contrast, motility (p = 0.012) and viability of spermatozoa (p = 0.007) were highly significant differences (p < 0.05 and p < 0.01) in dose 300 mg/kg BW. The testosterone levels have not significantly (p = 0.162; p > 0.05), but the tendency increased in dose 300 mg/kg BW (11.01%). Nevertheless, serum biochemistry was insignificant (p > 0.05) in all groups. Conclusion: The SE ameliorates fertility and serum biochemistry in DMPA-induced rats. [J Adv Vet Anim Res 2023; 10(1.000): 126-131]

Objective: The goal of this study was to look at quinolone-resistant (QR) Escherichia coli (E. coli) from retail beef and poultry meat in Egypt by looking at the QR mechanisms in the resistant strains. Materials and Methods: In total, 120 samples of raw poultry meat (n = 60) and beef meat (n = 60) were purchased from Mansoura retail stores between January and March 2021, and evaluated microbiologically for E. coli. Then, an antimicrobial sensitivity test was applied to all isolates. The prevalence of QR E. coli with concern for the QR determinants, including quinolone resistance-determining regions (QRDRs) mutations, the plasmid-mediated quinolone resistance gene (PMQR), and the efflux pump activity were determined. Results: The total prevalence of E. coli was 34.2% (41/120). Noticeably, the prevalence of E. coli in poultry meat (40%, 24/60) was higher than that of beef (28%, 17/60). All strains were assessed for their antimicrobial susceptibility using the disc diffusion technique; the highest rate of resistance (100%) was displayed to clindamycin and cefuroxime, followed by ampicillin (97.6%), doxycycline (92.7%), amoxicillin-clavulanate (92.7%), nalidixic acid (NA) (80.5%), sulfamethoxazole/trimethoprim (70.7%), chloramphenicol (63.4%), gentamicin, and azithromycin (58.5% each). Multiple antimicrobial resistance (strains resistant to three or more antimicrobial classes) was displayed by 97.6% of E. coli isolates. Regarding QR, 37 isolates could resist at least one of the examined quinolones. Regarding PMQR genes, qnrS was determined in 70% (7/10) of QR E. coli, while qnrA, qnrB, and qnrD were not identified. While the mutations determined regions of QR in the resistant E. coli isolates, S83L was the most prevalent in gyrase subunit A either alone or combined with D87N and D87Y, and three isolates of QR E. coli isolates revealed a topoisomerase IV subunit mutation harboring S80I. 20% of the isolates displayed efflux activity, as NA showed a considerable difference between its zones of inhibition. Conclusion: The high prevalence of antimicrobial-resistant E. coli, with concern for QR strains harboring different resistance mechanisms in poultry meat and beef, threatens the public’s health. Thus, standard manufacturing procedures and adequate hygiene conditions must be followed in all phases of meat preparation, production, and consumption, and public knowledge should be improved.   J. Adv. Vet. Anim. Res., 10(3): 490–499, September 2023 http://doi.org/10.5455/javar.2023.j702

Objective: The goal of this study was to look at quinolone-resistant (QR) Escherichia coli (E. coli) from retail beef and poultry meat in Egypt by looking at the QR mechanisms in the resistant strains. Materials and Methods: In total, 120 samples of raw poultry meat (n = 60) and beef meat (n = 60) were purchased from Mansoura retail stores between January and March 2021, and evaluated microbiologically for E. coli. Then, an antimicrobial sensitivity test was applied to all isolates. The prevalence of QR E. coli with concern for the QR determinants, including quinolone resistance-determining regions (QRDRs) mutations, the plasmid-mediated quinolone resistance gene (PMQR), and the efflux pump activity were determined. Results: The total prevalence of E. coli was 34.2% (41/120). Noticeably, the prevalence of E. coli in poultry meat (40%, 24/60) was higher than that of beef (28%, 17/60). All strains were assessed for their antimicrobial susceptibility using the disc diffusion technique; the highest rate of resistance (100%) was displayed to clindamycin and cefuroxime, followed by ampicillin (97.6%), doxycycline (92.7%), amoxicillin-clavulanate (92.7%), nalidixic acid (NA) (80.5%), sulfamethoxazole/trimethoprim (70.7%), chloramphenicol (63.4%), gentamicin, and azithromycin (58.5% each). Multiple antimicrobial resistance (strains resistant to three or more antimicrobial classes) was displayed by 97.6% of E. coli isolates. Regarding QR, 37 isolates could resist at least one of the examined quinolones. Regarding PMQR genes, qnrS was determined in 70% (7/10) of QR E. coli, while qnrA, qnrB, and qnrD were not identified. While the mutations determined regions of QR in the resistant E. coli isolates, S83L was the most prevalent in gyrase subunit A either alone or combined with D87N and D87Y, and three isolates of QR E. coli isolates revealed a topoisomerase IV subunit mutation harboring S80I. 20% of the isolates displayed efflux activity, as NA showed a considerable difference between its zones of inhibition. Conclusion: The high prevalence of antimicrobial-resistant E. coli, with concern for QR strains harboring different resistance mechanisms in poultry meat and beef, threatens the public's health. Thus, standard manufacturing procedures and adequate hygiene conditions must be followed in all phases of meat preparation, production, and consumption, and public knowledge should be improved.

Objective: Whole genome sequencing (WGS) of Aeromonas veronii Alim_AV_1000 isolated from ulcerative lesions of Shing fish (stringing catfish; Heteropneustes fossilis) was performed during the outbreak year 2021. Materials and Methods: Using next-generation sequencing (Illumina) technology, WGS was accomplished, resulting in the sequencing, assembly, and analysis of the entire genome of the A. veronii strain. Moreover, the genomic features, virulence factors, antimicrobial resistome, and phylogenetic analysis for the molecular evolution of this strain were also examined. Results: The genome size of the A. veronii Alim_AV_1000 strain was 4,494,515 bp, with an average G+C content of 58.87%. Annotation revealed the known transporters and genes linked to virulence, drug targets, and antimicrobial resistance. Conclusion: The findings of the phylogenetic analysis revealed that the strain of the present study has a close relationship with the China strain TH0426 and strain B56. This study provides novel information on A. veronii isolated from Shing fish in Bangladesh. J. Adv. Vet. Anim. Res., 10(3): 570–578, September 2023 http://doi.org/10.5455/javar.2023.j711

Objective: Whole genome sequencing (WGS) of Aeromonas veronii Alim_AV_1000 isolated from ulcerative lesions of Shing fish (stringing catfish; Heteropneustes fossilis) was performed during the outbreak year 2021. Materials and Methods: Using next-generation sequencing (Illumina) technology, WGS was accomplished, resulting in the sequencing, assembly, and analysis of the entire genome of the A. veronii strain. Moreover, the genomic features, virulence factors, antimicrobial resistome, and phylogenetic analysis for the molecular evolution of this strain were also examined. Results: The genome size of the A. veronii Alim_AV_1000 strain was 4,494,515 bp, with an average G+C content of 58.87%. Annotation revealed the known transporters and genes linked to virulence, drug targets, and antimicrobial resistance. Conclusion: The findings of the phylogenetic analysis revealed that the strain of the present study has a close relationship with the China strain TH0426 and strain B56. This study provides novel information on A. veronii isolated from Shing fish in Bangladesh.

Objective: The aerolysin (aerA) is a virulence indicator used to identify the pathogenicity of the Aeromonas strain. Targeting a pathogen’s crucial virulence gene for detection is essential, as it determines the potential threat to the host. This study aimed to develop a gold nanoparticle (AuNP) probe for detecting the gene aerA in Aeromonas hydrophila among field samples. Materials and Methods: Kidney samples among both healthy and sick Nile tilapias in five provinces of Luzon Island were collected for bacterial analysis. Screening using specific primers targeting aerA was conducted in parallel with testing the AuNPs probe on the same sample set. The positive control provided by BFAR-NFLD, confirmed by polymerase chain reaction (PCR) assay, was used as a positive sample containing the target gene. Results: The AuNP probe demonstrated a computed accuracy of 81.32%, sensitivity of 100%, and specificity of 81.26%. Among the 257 reactions, 59 were false positives, while no false negative results were observed. The AuNP probe could detect aerA at levels as low as 30 ng/µl. The low prevalence of the target gene may be attributed to the use of general media instead of specific media like Rimler-Shotts agar. Conclusion: The established colorimetric detection method for A. hydrophila with the aerA gene offers a swift alternative to PCR, negating the requirement for advanced equipment like a thermal cycler. J. Adv. Vet. Anim. Res., 10(4): 593–598, December 2023 http://doi.org/10.5455/javar.2023.j714

Objective: The aerolysin (aerA) is a virulence indicator used to identify the pathogenicity of the Aeromonas strain. Targeting a pathogen's crucial virulence gene for detection is essential, as it determines the potential threat to the host. This study aimed to develop a gold nanoparticle (AuNP) probe for detecting the gene aerA in Aeromonas hydrophila among field samples. Materials and Methods: Kidney samples among both healthy and sick Nile tilapias in five provinces of Luzon Island were collected for bacterial analysis. Screening using specific primers targeting aerA was conducted in parallel with testing the AuNPs probe on the same sample set. The positive control provided by BFAR-NFLD, confirmed by polymerase chain reaction (PCR) assay, was used as a positive sample containing the target gene. Results: The AuNP probe demonstrated a computed accuracy of 81.32%, sensitivity of 100%, and specificity of 81.26%. Among the 257 reactions, 59 were false positives, while no false negative results were observed. The AuNP probe could detect aerA at levels as low as 30 ng/μl. The low prevalence of the target gene may be attributed to the use of general media instead of specific media like Rimler-Shotts agar. Conclusion: The established colorimetric detection method for A. hydrophila with the aerA gene offers a swift alternative to PCR, negating the requirement for advanced equipment like a thermal cycler. [J Adv Vet Anim Res 2023; 10(4.000): 593-598]

Objective: The following study examines for the first time the changes that occur in the post-partum period following abortion in the first trimester of dairy cows using hormonal, hematological, and oxidant/antioxidant profiles. In addition, a bacteriological examination was also performed to explore the role of infections in the complications that occur during this period. Materials and Methods: One hundred cows were split into two equal groups: The first group enrolled cows that suffered from abortion in the first trimester. The second group enrolled cows that did not experience abortion problems (the control group). Uterine swabs were collected from cows. Blood samples were collected for hormonal, hematological, and oxidative profiles. Results: Results reveal that Escherichia coli, Staphylococcus spp., and Streptococcus spp. are the opportunistic bacteria that were isolated from abortive cows with multidrug-resistant (MDR) characteristics. Red blood cell (RBC) count, hemoglobin, mean corpuscular hemoglobin (MCH), and MCH concentration (MCHC) were significantly higher in the abortive group than in controls in the first 3 days after calving. Conversely, total leukocyte count, platelet count, neutrophils, eosinophils, and immunoglobulin G and M were significantly lower in the abortion group than in controls. The concentrations of estradiol, prostaglandin F2α, oxytocin, and cortisol are significantly increased in the abortive cows, while progesterone is significantly decreased. The levels of malondialdehyde (MDA) were higher in the abortive group, while the levels of superoxide dismutase (SOD), glutathione peroxidase (GPx), and total antioxidant capacity (TAC) were lower. Conclusion: Abortion during the first trimester of pregnancy increases the risk of postpartum opportunistic bacterial invasion of the uterus. Oxidative stress (OS) and neutropenia are the most important findings that may occur in the postpartum period after abortion and may be due to the abortion itself or its predisposition to opportunistic bacterial invasion of the uterus, which finally causes a fertility reduction. J. Adv. Vet. Anim. Res., 10(4): 654–666, December 2023 http://doi.org/10.5455/javar.2023.j721

Objective: The following study examines for the first time the changes that occur in the post-partum period following abortion in the first trimester of dairy cows using hormonal, hematological, and oxidant/antioxidant profiles. In addition, a bacteriological examination was also performed to explore the role of infections in the complications that occur during this period. Materials and Methods: One hundred cows were split into two equal groups: The first group enrolled cows that suffered from abortion in the first trimester. The second group enrolled cows that did not experience abortion problems (the control group). Uterine swabs were collected from cows. Blood samples were collected for hormonal, hematological, and oxidative profiles. Results: Results reveal that Escherichia coli, Staphylococcus spp., and Streptococcus spp. are the opportunistic bacteria that were isolated from abortive cows with multidrug-resistant (MDR) characteristics. Red blood cell (RBC) count, hemoglobin, mean corpuscular hemoglobin (MCH), and MCH concentration (MCHC) were significantly higher in the abortive group than in controls in the first 3 days after calving. Conversely, total leukocyte count, platelet count, neutrophils, eosinophils, and immunoglobulin G and M were significantly lower in the abortion group than in controls. The concentrations of estradiol, prostaglandin F2α, oxytocin, and cortisol are significantly increased in the abortive cows, while progesterone is significantly decreased. The levels of malondialdehyde (MDA) were higher in the abortive group, while the levels of superoxide dismutase (SOD), glutathione peroxidase (GPx), and total antioxidant capacity (TAC) were lower. Conclusion: Abortion during the first trimester of pregnancy increases the risk of postpartum opportunistic bacterial invasion of the uterus. Oxidative stress (OS) and neutropenia are the most important findings that may occur in the postpartum period after abortion and may be due to the abortion itself or its predisposition to opportunistic bacterial invasion of the uterus, which finally causes a fertility reduction. [J Adv Vet Anim Res 2023; 10(4.000): 654-666]

Objective: Cadmium chloride (CdCl2 ) is an environmentally toxic pollutant that can cause reprotoxicity. Cyperus esculentus and Euterpe oleracea are potent antioxidant plants currently used to counteract the action of harmful pollutants. The present experiment was intended to evaluate and comp are the role of C. esculentus hydroethanolic extract (CHE) and E. oleracea in treating the reprotoxicity induced by CdCl2 in rats. Materials and Methods: Forty adult male rats (160–210 gm) were allocated into five groups equally. Control group: received 5 ml of normal saline (NS); the other treatment groups were injected with CdCl2 as a single dose for two weeks to induce testicular toxicity. After 14 days, the four groups were treated orally daily for two months as follows: The cadmium group (Cd) received NS, the third group (TC) was administered 800 mg/kg BW of CHE, the fourth group (TO) received 500 mg/kg BW of E. oleracea, and the fifth group (TCO) received CHE with E. oleracea. Results: The live sperm and motility, serum testosterone, follicle-stimulating hormone (FSH), testicular superoxide dismutase (SOD), catalase (CAT), steroidogenic acute regulatory protein (StAR), 17β-hydroxysteroid dehydrogenase, and 3β-hydroxysteroid dehydrogenase (3β-HSD) were significantly increased in the TCO, TC, and TO groups compared with the Cd group. Testicular nitric oxide and malondialdehyde were elevated significantly in the Cd group compared to the TC, TO, TCO, and control groups. The fold changes of Fshβ, Lhβ, and Gnrh genes were upregulated in the TCO group compared to the Cd and control groups. Conclusion: The combination of CHE with E. oleracea showed improvements in rat testicles affected by cadmium toxicity via upregulated reproductive gene expression and its antioxidant effects. J. Adv. Vet. Anim. Res., 10(4): 685–695, December 2023 http://doi.org/10.5455/javar.2023.j724

Objective: Cadmium chloride (CdCl2) is an environmentally toxic pollutant that can cause reprotoxicity. Cyperus esculentus and Euterpe oleracea are potent antioxidant plants currently used to counteract the action of harmful pollutants. The present experiment was intended to evaluate and comp are the role of C. esculentus hydroethanolic extract (CHE) and E. oleracea in treating the reprotoxicity induced by CdCl2 in rats. Materials and Methods: Forty adult male rats (160–210 gm) were allocated into five groups equally. Control group: received 5 ml of normal saline (NS); the other treatment groups were injected with CdCl2 as a single dose for two weeks to induce testicular toxicity. After 14 days, the four groups were treated orally daily for two months as follows: The cadmium group (Cd) received NS, the third group (TC) was administered 800 mg/kg BW of CHE, the fourth group (TO) received 500 mg/kg BW of E. oleracea, and the fifth group (TCO) received CHE with E. oleracea. Results: The live sperm and motility, serum testosterone, follicle-stimulating hormone (FSH), testicular superoxide dismutase (SOD), catalase (CAT), steroidogenic acute regulatory protein (StAR), 17β-hydroxysteroid dehydrogenase, and 3β-hydroxysteroid dehydrogenase (3β-HSD) were significantly increased in the TCO, TC, and TO groups compared with the Cd group. Testicular nitric oxide and malondialdehyde were elevated significantly in the Cd group compared to the TC, TO, TCO, and control groups. The fold changes of Fshβ, Lhβ, and Gnrh genes were upregulated in the TCO group compared to the Cd and control groups. Conclusion: The combination of CHE with E. oleracea showed improvements in rat testicles affected by cadmium toxicity via upregulated reproductive gene expression and its antioxidant effects. [J Adv Vet Anim Res 2023; 10(4.000): 685-695]

Objective: The principal objective of this research was to examine the influence of zinc-methionine (Zn-Met) addition on feed on the in vitro ruminal enzyme activities, fermentation characteristics, methane production, and digestibilities of feed nutrients. Materials and Methods: The dosage of Zn-Met as a source of organic Zn was added to feed based on dry matter (DM) as follows: 0-without Zn addition (CON), 30 Zn mg/kg-low (LZM), 60 Zn mg/ kg-medium (MZM), and 90 Zn mg/kg-high (HZM). Results: The results indicated significant impacts of Zn-Met addition on various parameters. Compared to the CON group, all Zn-Met treatments resulted in increased total volatile fatty acids (VFA) (linear; p < 0.001), carboxymethyl-cellulase activity (linear; p < 0.001), and gas production at 48-h of incubation (linear; p < 0.001, quadratic; p < 0.001). Additionally, the MZM and HZM groups exhibited higher levels of amylase activity (linear; p < 0.001), protease activity (linear; p = 0.006), microbial protein (linear; p = 0.009), DM digestibility (linear; p < 0.001), organic matter (OM) digestibility (linear; p < 0.001), crude protein (CP) digestibility (linear; p = 0.004), and crude fiber (CF) digestibility (linear; p = 0.003) than CON treatment. However, the treatments did not have any noteworthy effects on pH, the individual VFA (acetate, propionate, and butyrate) proportions, NH3 -N concentration, and methane production (p > 0.05). Conclusion: It could be summarized that supplementing 60 and 90 Zn mg/kg DM as Zn-Met could improve the in vitro ruminal enzyme activities, fermentation characteristics, and nutrient digestibility without affecting methane production. J. Adv. Vet. Anim. Res., 10(4): 696–703, December 2023 http://doi.org/10.5455/javar.2023.j725

Objective: The principal objective of this research was to examine the influence of zinc-methionine (Zn-Met) addition on feed on the in vitro ruminal enzyme activities, fermentation characteristics, methane production, and digestibilities of feed nutrients. Materials and Methods: The dosage of Zn-Met as a source of organic Zn was added to feed based on dry matter (DM) as follows: 0-without Zn addition (CON), 30 Zn mg/kg-low (LZM), 60 Zn mg/kg-medium (MZM), and 90 Zn mg/kg-high (HZM). Results: The results indicated significant impacts of Zn-Met addition on various parameters. Compared to the CON group, all Zn-Met treatments resulted in increased total volatile fatty acids (VFA) (linear; p < 0.001), carboxymethyl-cellulase activity (linear; p < 0.001), and gas production at 48-h of incubation (linear; p < 0.001, quadratic; p < 0.001). Additionally, the MZM and HZM groups exhibited higher levels of amylase activity (linear; p < 0.001), protease activity (linear; p = 0.006), microbial protein (linear; p = 0.009), DM digestibility (linear; p < 0.001), organic matter (OM) digestibility (linear; p < 0.001), crude protein (CP) digestibility (linear; p = 0.004), and crude fiber (CF) digestibility (linear; p = 0.003) than CON treatment. However, the treatments did not have any noteworthy effects on pH, the individual VFA (acetate, propionate, and butyrate) proportions, NH3-N concentration, and methane production (p > 0.05). Conclusion: It could be summarized that supplementing 60 and 90 Zn mg/kg DM as Zn-Met could improve the in vitro ruminal enzyme activities, fermentation characteristics, and nutrient digestibility without affecting methane production. [J Adv Vet Anim Res 2023; 10(4.000): 696-703]

Objective: To determine the correlation between the molecular weight (MW) of proteins in seminal plasma and spermatozoa and the quality of fresh and frozen semen production in Pasundan bulls. Materials and methods: Nine selected Pasundan bulls, aged 5–10 years, from the Regional Artificial Insemination Center at Ciamis, West Java, Indonesia, were used in the study, with fresh semen sperm motility ≥70% and <70%. We analyzed the motility, viability, integrity of the intact plasma membrane (IPM), and the morphological characteristics of spermatozoa. 1D-SDS-PAGE analysis was performed to determine the protein profile by assessing MW, depicted as bands on the gel. Results: The motility, viability, and IPM of spermatozoa had lower values (p < 0.05) in Pasundan bulls named Bagaskara and Kertarajasa compared to the other bulls. Proteins with MW 35–50 kDa were not detected in the seminal plasma of Pasundan bulls, exhibiting low quality in fresh semen. The correlation analysis showed that the non-detected proteins with MW 35–50 kDa in seminal plasma correlated with spermatozoa motility (r = 0.421), viability (r = 0.424), and IPM (r = 0.428) so that fresh semen quality was low in both Pasundan bulls. Analysis of semen volume, spermatozoa concentration, and spermatozoa motility showed that the average frozen semen production of Pasundan bulls per ejaculate was 128.73 ± 15.35 straws. Conclusion: Protein analysis based on MW is a predictive indicator for the quality of fresh semen and the production of frozen semen in Pasundan bulls. Evaluation parameters of fresh semen quality by MW analysis can be used to select Pasundan bulls in Indonesia. J. Adv. Vet. Anim. Res., 10(4): 730–737, December 2023 http://doi.org/10.5455/javar.2023.j728

Objective: To determine the correlation between the molecular weight (MW) of proteins in seminal plasma and spermatozoa and the quality of fresh and frozen semen production in Pasundan bulls. Materials and methods: Nine selected Pasundan bulls, aged 5–10 years, from the Regional Artificial Insemination Center at Ciamis, West Java, Indonesia, were used in the study, with fresh semen sperm motility ≥70% and [J Adv Vet Anim Res 2023; 10(4.000): 730-737]