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]

Objective: The aim of this study is to detect the prevalence and diagnosis of molecular characteristics of Cryptosporidium infection in handlers of domestic pigeons in Baghdad City. Traditional and molecular diagnostic methods were employed to detect and identify Cryptosporidium species. Materials and methods: Sixty stool samples were collected from the handlers of domestic pigeons, and various techniques, including direct smear, flotation concentration, staining methods, and DNA extraction coupled with nested polymerase chain reaction (PCR), were utilized. Results: The results obtained from traditional methods indicated an overall infection rate of 55% in handlers of domestic pigeons, while significant variations were noted between male and female handlers. Age group 21–40 years were found to have higher infection rates were found to have higher infection rates. The prevalence of Cryptosporidium also displayed temporal variability throughout the study period. Molecular analysis using nested PCR revealed higher infection rates of 86% in handlers of domestic pigeon samples. Genotyping and phylogenetic analysis identified the presence of Cryptosporidium parvam species in handlers of domestic pigeons, indicating zoonotic transmission potential. Conclusion: These findings underscore the high prevalence of Cryptosporidium infection among handlers of domestic pigeons in Baghdad City. J. Adv. Vet. Anim. Res., 10(4): 744–750, December 2023 http://doi.org/10.5455/javar.2023.j730

Objective: The aim of this study is to detect the prevalence and diagnosis of molecular characteristics of Cryptosporidium infection in handlers of domestic pigeons in Baghdad City. Traditional and molecular diagnostic methods were employed to detect and identify Cryptosporidium species. Materials and methods: Sixty stool samples were collected from the handlers of domestic pigeons, and various techniques, including direct smear, flotation concentration, staining methods, and DNA extraction coupled with nested polymerase chain reaction (PCR), were utilized. Results: The results obtained from traditional methods indicated an overall infection rate of 55% in handlers of domestic pigeons, while significant variations were noted between male and female handlers. Age group 21–40 years were found to have higher infection rates were found to have higher infection rates. The prevalence of Cryptosporidium also displayed temporal variability throughout the study period. Molecular analysis using nested PCR revealed higher infection rates of 86% in handlers of domestic pigeon samples. Genotyping and phylogenetic analysis identified the presence of Cryptosporidium parvam species in handlers of domestic pigeons, indicating zoonotic transmission potential. Conclusion: These findings underscore the high prevalence of Cryptosporidium infection among handlers of domestic pigeons in Baghdad City. [J Adv Vet Anim Res 2023; 10(4.000): 744-750]

Objective: Protein and carbohydrates are substances needed by ruminants, especially sheep. Providing protein and carbohydrates must pay attention to their degradation. In addition, balancing nutrients to meet the nutritional needs of rumen microbes is very important because the unbalanced availability of rumen degradable protein (RDP) and readily available carbohydrate (RAC) at one time can cause suboptimal microbial protein synthesis efficiency. Materials and Methods: Completely randomized design with a nonfactorial pattern of five treatments with three replications. Treatment includes RDP and RAC ratios, namely R1 = 2.30, R2 = 2.00, R3 = 1.70, R4 = 1.50, and R5 = 1.30. Results: The results showed that the balance of RDP and RAC in sheep ration formulation in vitro had a very significant effect (p < 0.01) on NH3 , microbial protein synthesis, total gas, total microbes, and organic matter digestibility (OMD) but had no significant effect (p > 0.05) on rumen pH and dry matter digestibility (DMD). Conclusion: The optimal balance of RDP and RAC in the formulation of sheep rations in vitro was obtained at a ratio of 2.30 with NH3 (mM) 8.47, rumen pH 5.97, microbial protein synthesis (mg/100 ml) 123, gas (ml/g of material) 145, total microbes (cells/ml) 2.012 × 106 , (log CFU cells/ ml) 6.3025, DMD 61.0%, and OMD 63.1%. J. Adv. Vet. Anim. Res., 10(4): 738–743, December 2023 http://doi.org/10.5455/javar.2023.j729

Objective: Protein and carbohydrates are substances needed by ruminants, especially sheep. Providing protein and carbohydrates must pay attention to their degradation. In addition, balancing nutrients to meet the nutritional needs of rumen microbes is very important because the unbalanced availability of rumen degradable protein (RDP) and readily available carbohydrate (RAC) at one time can cause suboptimal microbial protein synthesis efficiency. Materials and Methods: Completely randomized design with a nonfactorial pattern of five treatments with three replications. Treatment includes RDP and RAC ratios, namely R1 = 2.30, R2 = 2.00, R3 = 1.70, R4 = 1.50, and R5 = 1.30. Results: The results showed that the balance of RDP and RAC in sheep ration formulation in vitro had a very significant effect (p < 0.01) on NH3, microbial protein synthesis, total gas, total microbes, and organic matter digestibility (OMD) but had no significant effect (p > 0.05) on rumen pH and dry matter digestibility (DMD). Conclusion: The optimal balance of RDP and RAC in the formulation of sheep rations in vitro was obtained at a ratio of 2.30 with NH3 (mM) 8.47, rumen pH 5.97, microbial protein synthesis (mg/100 ml) 123, gas (ml/g of material) 145, total microbes (cells/ml) 2.012 × 106, (log CFU cells/ml) 6.3025, DMD 61.0%, and OMD 63.1%. [J Adv Vet Anim Res 2023; 10(4.000): 738-743]

Objective: This study evaluated the utilization of marigold leaves (MGLs) in rations and their impact on rumen enzyme activity, fermentation parameters, methane (CH4 ) emission, and nutrient digestibility in vitro. Materials and Methods: The experimental diets comprised different proportions of MGL incorporated into the dry matter (DM) rations. Experimental design: The MGL treatments in diets include 0% (MGL-0), 7% (MGL-7), and 14% (MGL-14). Results: Results indicated that MGL-14 substantially raised (p < 0.05) the rumen parameters, including NH3 –N and microbial protein, total volatile fatty acids, acetate (C2 ), propionate (C3 ), butyrate (C4 ), and the C2 :C3 ratio. In contrast, the MGL-7 and MGL-14 groups experienced a noteworthy reduction (p < 0.05) in the total protozoa population. The MGL-7 and MGL-14 treatments also led to a substantial increase in the digestibility of DM, organic matter (OM), and crude fiber (CF) in the rumen. However, they also resulted in a decline (p < 0.05) in crude protein (CP) digestibility. The DM and OM total digestibilities were higher (p < 0.05) in the MGL-14 and MGL-7 groups. The utilization of MGL did not influence (p > 0.05) the ruminal enzyme activities (carboxymethyl cellulase, amylase, protease), cumulative gas production, kinetics, ruminal pH value, CH4 and CO2 production, total CF, and CP digestibility. Conclusion: The utilization of MGL until 14% DM in diets can enhance ruminal fermentation parameters and nutrient digestibility in vitro without negatively affecting gas production kinetics or ruminal enzyme activities. However, it did not have any impact on CH4 production. J. Adv. Vet. Anim. Res., 10(4): 782–793, December 2023 http://doi.org/10.5455/javar.2023.j734

Objective: This study evaluated the utilization of marigold leaves (MGLs) in rations and their impact on rumen enzyme activity, fermentation parameters, methane (CH4) emission, and nutrient digestibility in vitro. Materials and Methods: The experimental diets comprised different proportions of MGL incorporated into the dry matter (DM) rations. Experimental design: The MGL treatments in diets include 0% (MGL-0), 7% (MGL-7), and 14% (MGL-14). Results: Results indicated that MGL-14 substantially raised (p < 0.05) the rumen parameters, including NH3–N and microbial protein, total volatile fatty acids, acetate (C2), propionate (C3), butyrate (C4), and the C2:C3 ratio. In contrast, the MGL-7 and MGL-14 groups experienced a noteworthy reduction (p < 0.05) in the total protozoa population. The MGL-7 and MGL-14 treatments also led to a substantial increase in the digestibility of DM, organic matter (OM), and crude fiber (CF) in the rumen. However, they also resulted in a decline (p < 0.05) in crude protein (CP) digestibility. The DM and OM total digestibilities were higher (p < 0.05) in the MGL-14 and MGL-7 groups. The utilization of MGL did not influence (p > 0.05) the ruminal enzyme activities (carboxymethyl cellulase, amylase, protease), cumulative gas production, kinetics, ruminal pH value, CH4 and CO2 production, total CF, and CP digestibility. Conclusion: The utilization of MGL until 14% DM in diets can enhance ruminal fermentation parameters and nutrient digestibility in vitro without negatively affecting gas production kinetics or ruminal enzyme activities. However, it did not have any impact on CH4 production. [J Adv Vet Anim Res 2023; 10(4.000): 782-793]