Objective: This research was arranged to explore the effect of supplementation of a combination of Lactobacillus plantarum and Saccharomyces cerevisiae as a new probiotic in fermented rice straw-based rations on in vitro digestibility and ruminal characteristics. Materials and Methods: A randomized group design with 3 types of treatment and 4 replications as a group was used in this study. A probiotic inoculum containing L. plantarum and S. cerevisiae with 1 × 1010 colony-forming unit (CFU)/ml. Treatments were followed by: P1 = complete rations without probiotics (control), P2 = P1 supplemented 0.5% probiotics, and P3 = P1 supplemented 1% probiotics. Substrate complete rations were based on the fermented rice straw and concentrate (60%:40%). Parameters of digestibility and rumen fermentation products were determined after 48 h of incubation. Results: Probiotics supplemented with fermented rice straw-based rations significantly increased (p < 0.05) digestibility and rumen characteristics in vitro. Supplementation with 1% probiotics (P3) produces the highest digestibility compared to other treatments: in-vitro dry matter digestibility (IVDMD) (55%), in-vitro organic matter digestibility (IVOMD) (58.28%), in-vitro crude protein digestibility (IVCPD) (84.42%), in-vitro acid detergent fiber digestibility (IVADFD) (53.99%), in-vitro neutral detergent fiber digestibility (IVNDFD) (58.39%), and in-vitro cellulose digestibility (IVCLD) (67.12%). Rumen pH (6.76–6.80) did not change significantly (p > 0.05) due to supplemented probiotics. Probiotic supplementation in rations significantly (p < 0.05) increased the content of NH3 and total volatile fatty acid (VFA). Supplementation with 1% probiotic (P3) showed the highest concentration of NH3 (26.56 mg/100 ml) and was also followed by the total VFA (115.75 mM) compared to the control (22.59 mg/100 ml and 103.00 mM, respectively). Conclusion: Supplementation of 1% probiotics (combination of L. plantarum and S. cerevisiae) containing 1 × 1010 CFU/ml in fermented rice straw-based rations increases nutrient digestibility, that is, IVDMD, IVOMD, IVCPD, IVADFD, IVNDFD, and IVCLD, and also increases rumen fermentation, which is the concentration of NH3 and total VFA.      J. Adv. Vet. Anim. Res., 10(1): 96–102, March 2023 http://doi.org/10.5455/javar.2023.j657

Objective: This research was arranged to explore the effect of supplementation of a combination of Lactobacillus plantarum and Saccharomyces cerevisiae as a new probiotic in fermented rice straw-based rations on in vitro digestibility and ruminal characteristics. Materials and Methods: A randomized group design with 3 types of treatment and 4 replications as a group was used in this study. A probiotic inoculum containing L. plantarum and S. cerevisiae with 1 × 1010 colony-forming unit (CFU)/ml. Treatments were followed by: P1 = complete rations without probiotics (control), P2 = P1 supplemented 0.5% probiotics, and P3 = P1 supplemented 1% probiotics. Substrate complete rations were based on the fermented rice straw and concen¬trate (60%:40%). Parameters of digestibility and rumen fermentation products were determined after 48 h of incubation. Results: Probiotics supplemented with fermented rice straw-based rations significantly increased (p < 0.05) digestibility and rumen characteristics in vitro. Supplementation with 1% probiotics (P3) produces the highest digestibility compared to other treatments: in-vitro dry matter digestibil¬ity (IVDMD) (55%), in-vitro organic matter digestibility (IVOMD) (58.28%), in-vitro crude protein digestibility (IVCPD) (84.42%), in-vitro acid detergent fiber digestibility (IVADFD) (53.99%), in-vitro neutral detergent fiber digestibility (IVNDFD) (58.39%), and in-vitro cellulose digestibility (IVCLD) (67.12%). Rumen pH (6.76–6.80) did not change significantly (p > 0.05) due to supplemented probiotics. Probiotic supplementation in rations significantly (p < 0.05) increased the content of NH3 and total volatile fatty acid (VFA). Supplementation with 1% probiotic (P3) showed the highest concentration of NH3 (26.56 mg/100 ml) and was also followed by the total VFA (115.75 mM) compared to the control (22.59 mg/100 ml and 103.00 mM, respectively). Conclusion: Supplementation of 1% probiotics (combination of L. plantarum and S. cerevisiae) containing 1 × 1010 CFU/ml in fermented rice straw-based rations increases nutrient digestibility, that is, IVDMD, IVOMD, IVCPD, IVADFD, IVNDFD, and IVCLD, and also increases rumen fermenta¬tion, which is the concentration of NH3 and total VFA. [J Adv Vet Anim Res 2023; 10(1.000): 96-102]

Objective: This work examined the results of different dietary protein levels on indigenous Nigerian guinea fowl growth performance and carcass yield. Materials and Methods: One hundred and eight unsexed indigenous keets were randomly assigned to three treatments with experimental diets containing 22%, 24%, and 26% crude protein (CP) in a completely randomized design, with 3 replicates with 12 keets in each replicate. The parameters were measured, including proximate composition, feed intake, growth rate, feed conversion ratio (FCR), and carcass yield. Results: The birds fed diets containing 24% CP gained the most body weight at 619.83 gm and had the highest FCR of 3.45. The presence of CP had an impact that was significant (p < 0.05) on most carcass traits and prime cuts but not on gastrointestinal tract weight, head weight, dress percentage, or organ yield. Conclusion: The study concluded that the dietary CP level of 24% was optimal for body weight gain and carcass yield. J. Adv. Vet. Anim. Res., 10(3): 403–409, September 2023 http://doi.org/10.5455/javar.2023.j693

Objective: This work examined the results of different dietary protein levels on indigenous Nigerian guinea fowl growth performance and carcass yield. Materials and Methods: One hundred and eight unsexed indigenous keets were randomly assigned to three treatments with experimental diets containing 22%, 24%, and 26% crude protein (CP) in a completely randomized design, with 3 replicates with 12 keets in each replicate. The parameters were measured, including proximate composition, feed intake, growth rate, feed conversion ratio (FCR), and carcass yield. Results: The birds fed diets containing 24% CP gained the most body weight at 619.83 gm and had the highest FCR of 3.45. The presence of CP had an impact that was significant (p < 0.05) on most carcass traits and prime cuts but not on gastrointestinal tract weight, head weight, dress percentage, or organ yield. Conclusion: The study concluded that the dietary CP level of 24% was optimal for body weight gain and carcass yield.

Objective: The effect of zeolite on Ca plasma concentration, reproductive parameters, body condition score (BCS), and colostrum properties was evaluated in Holstein dairy cows during the postpartum period. Materials and Methods: Sixty pregnant cows were allocated to three experimental groups, including 1) a control (CON) diet; 2) a CON diet + 0.75% dry matter (DM) zeolite (100 gm/day/ cow); and 3) a CON diet + 1.5% DM zeolite (200 gm/day/cow). Experimental diets were fed to cows during the last 4 weeks of pregnancy. Blood sample evaluation for Ca concentration was done. The prevalence of hypocalcemia and reproductive parameters, including BCS and colostrum properties, was also measured. Results: Total Ca and ionized calcium (Ca++) concentrations in plasma at 6 and 12 h after calving were higher in zeolite-consuming cows than those in CON s (p < 0.01). Mean frequencies for severe and subclinical hypocalcemia in zeolite-consuming cows were 11.64% and 19.36% lower than those in the CON group, respectively (p < 0.01). The mean pregnancy efficiency of these cows was also 12.94% higher than the CON (p < 0.01). The mean BCS (p ≤ 0.05) and colostrum quality (p < 0.01) of zeolite-consuming cows were at maximum, but their mean colostrum yield was lower than that of the CON group (p < 0.01). Conclusion: Zeolite dietary supplementation at levels of 0.75% and 1.5% DM of the periparturient diet is recommended for better CON of hypocalcemia, improved BCS and colostrum quality, and better prevention of postpartum reproductive disorders in dairy cows. J. Adv. Vet. Anim. Res., 10(3): 410–420, September 2023 http://doi.org/10.5455/javar.2023.j694

Objective: The effect of zeolite on Ca plasma concentration, reproductive parameters, body condition score (BCS), and colostrum properties was evaluated in Holstein dairy cows during the postpartum period. Materials and Methods: Sixty pregnant cows were allocated to three experimental groups, including 1) a control (CON) diet; 2) a CON diet + 0.75% dry matter (DM) zeolite (100 gm/day/cow); and 3) a CON diet + 1.5% DM zeolite (200 gm/day/cow). Experimental diets were fed to cows during the last 4 weeks of pregnancy. Blood sample evaluation for Ca concentration was done. The prevalence of hypocalcemia and reproductive parameters, including BCS and colostrum properties, was also measured. Results: Total Ca and ionized calcium (Ca++) concentrations in plasma at 6 and 12 h after calving were higher in zeolite-consuming cows than those in CON s (p < 0.01). Mean frequencies for severe and subclinical hypocalcemia in zeolite-consuming cows were 11.64% and 19.36% lower than those in the CON group, respectively (p < 0.01). The mean pregnancy efficiency of these cows was also 12.94% higher than the CON (p < 0.01). The mean BCS (p ≤ 0.05) and colostrum quality (p < 0.01) of zeolite-consuming cows were at maximum, but their mean colostrum yield was lower than that of the CON group (p < 0.01). Conclusion: Zeolite dietary supplementation at levels of 0.75% and 1.5% DM of the preparturient diet is recommended for better CON of hypocalcemia, improved BCS and colostrum quality, and better prevention of postpartum reproductive disorders in dairy cows.

Objective: This study aimed to assess the effect of adding flavoring agents to fermented cassava pulp (FCPU) on nutrient utilization, ruminal fermentation characteristics, and blood metabolites in growing cattle. Materials and Methods: A duplicated 3 × 3 Latin square design was randomly assigned to six growing beef cattle. Treatments were: 1) untreated FCPU (control), 2) 0.05% w/v vanilla-flavored FCPU, and 3) 0.05% w/v amyl acetate-flavored FCPU. Results: The results showed that flavoring agents did not affect dry matter intake (DMI) or digestibility of nutrients. Rumen pH and ammonia nitrogen concentrations did not change all treatments post feeding. Flavoring added to FCPU after feeding did not affect total volatile fatty acid (VFA) or VFA percentage. However, propionic acid levels tended to be lower in the vanilla-flavored FCPU group than those in the unflavored FCPU group at 0 h post feeding (p < 0.01). Growing bulls fed vanilla-flavored FCPU tended to have greater fungal zoospores in the rumen than those fed amyl acetate-flavored FCPU (p < 0.1) at 2 h after feeding. Dietary treatments did not affect blood glucose and urea nitrogen concentrations (p > 0.05). However, blood triglyceride concentration was greater for cattle fed a control diet than other treatments at 0 h post feeding (p < 0.05) and tended to be higher than those fed vanilla-flavored FCPU at 2 h after feeding (p < 0.1). Conclusion: It was suggested that adding vanilla or amyl acetate flavor to the FCPU showed no adverse effects on ruminal fermentation, blood metabolites, or nutritional digestibility; however, it did not increase DMI. J. Adv. Vet. Anim. Res., 10(3): 437–448, September 2023 http://doi.org/10.5455/javar.2023.j697

Objective: This study aimed to assess the effect of adding flavoring agents to fermented cassava pulp (FCPU) on nutrient utilization, ruminal fermentation characteristics, and blood metabolites in growing cattle. Materials and Methods: A duplicated 3 × 3 Latin square design was randomly assigned to six growing beef cattle. Treatments were: 1) untreated FCPU (control), 2) 0.05% w/v vanilla-flavored FCPU, and 3) 0.05% w/v amyl acetate-flavored FCPU. Results: The results showed that flavoring agents did not affect dry matter intake (DMI) or digestibility of nutrients. Rumen pH and ammonia nitrogen concentrations did not change all treat¬ments postfeeding. Flavoring added to FCPU after feeding did not affect total volatile fatty acid (VFA) or VFA percentage. However, propionic acid levels tended to be lower in the vanilla-flavored FCPU group than those in the unflavored FCPU group at 0 h postfeeding (p < 0.01). Growing bulls fed vanilla-flavored FCPU tended to have greater fungal zoospores in the rumen than those fed amyl acetate-flavored FCPU (p < 0.1) at 2 h after feeding. Dietary treatments did not affect blood glucose and urea nitrogen concentrations (p > 0.05). However, blood triglyceride concentration was greater for cattle fed a control diet than other treatments at 0 h postfeeding (p < 0.05) and tended to be higher than those fed vanilla-flavored FCPU at 2 h afterfeeding (p < 0.1). Conclusion: It was suggested that adding vanilla or amyl acetate flavor to the FCPU showed no adverse effects on ruminal fermentation, blood metabolites, or nutritional digestibility; however, it did not increase DMI.

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]