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: This study sought to determine the occurrence, molecular identification, antimicrobi¬al-resistant trends, and gene distribution of Staphylococcus aureus in pet cats and their owners' hand swabs. Materials and Methods: From different places and clinics in Mymensingh and Dhaka, 168 pet cat samples and 42 hand swab samples from cat owners were obtained. The organisms were scruti¬nized by assessing the outcomes using conventional and molecular techniques. The disc diffusion technique was applied to find the resistance pattern against 12 antibiotics, and genes were dis¬covered by targeting specific genes using PCR. Results: The occurrence of pathogenic S. aureus in pet cats was 7.74%, while it was 9.50% in pet owners' hand swabs, and 25.0% of the pet owner's hand swabs contained these genes. Staphylococcus aureus was utterly resistant to amoxicillin, ampicillin, cefixime, erythromycin, and imipenem in both pet cat and hand swabs of pet owner samples. All S. aureus isolates had a multi¬drug-resistant phenotype, and 1 from pet cats (O19) and 1 from pet owner hand swabs (H9) were resistant to all 12 antibiotics in the 7 antimicrobial classes. Several antibiotic-resistance genes were detected by PCR. Conclusion: The study confirmed multidrug-resistant pathogenic S. aureus in pet cats and their owners in Bangladesh, indicating a major health risk to both people and cats. Thus, a holistic and integrated one-health approach between veterinary and medical specialists is needed to mitigate the global distribution of these zoonotic antibiotic-resistant S. aureus strains.

Objective: Estimating microbial protein synthesis (MPS) in the rumen of growing lambs based on the urinary excretion of purine derivatives (PDs) and forage to concentrate (F/C) ratio. Materials and Methods: 36 similar-growing male lambs (weight 32.53 ± 1.90 kg; age 93 ± 6.63 days) were used in a completely randomized design with four groups: a) the 20–80 F/C ratio (dry hay 10% + wheat straw 10%), b) the 20–80 F/C ratio (dry hay 0% + wheat straw 20%), c) the 10–90 F/C ratio (dry hay 5% + wheat straw 5%), d) and the 10–90 F/C ratio (dry hay 0% + wheat straw 10%) with nine replicates. Results: Total PD and rumen MPS synthesized increased (10.98 vs. 13.25 mmol/day and 59.45 vs. 71.80 gm/day) in group d compared to group a. Dry organic matter intake (0.869 kg/day), fermentable dry organic matter (0.563 kg/day), and microbial nitrogen (N) yield (11.48 gm/day) of group d were at the maximum, but in terms of gN/kg dry organic matter (22.37 gm/kg), the mean of group c was higher than others. Conclusion: Increasing the level of food concentrate and the gradual removal of alfalfa from the diet increased the excretion of PD and MPS in the rumen. It was also found that urinary PD monitoring is an accurate indicator for the estimation of MPS.

Objective: The study aimed to determine the potential anthelmintic activity of the ethyl acetate extract of Eleusine indica that will result in an effective reduction in fecal egg per gram (EPG) counts in naturally infected goats compared to the commercial anthelmintic levamisole. Materials and Methods: The experimental animals were 21 goats naturally infected with gas¬trointestinal nematodes. The goats were divided into groups that were given a single dose of E. indica extract. Five concentrations of E. indica were tested for anthelmintic activity: 100, 200, 300, 400, and 500 mg extract/kg body weight. Fecal sample collection was done before treatment, during the first treatment, and every week thereafter for 28 days post-treatment (dpt). A modi¬fied McMaster technique was used to determine the EPG of feces, and the mean efficacies of the extracts were compared with those of the commercial anthelmintic levamisole. Results: As early as 7 dpt, there was an observed reduction in the epg counts after the administration of E. indica extracts across all concentrations. Administering 500 mg of extract/kg body weight resulted in a maximum efficacy of 56.21%. However, the efficacy achieved was lower than that of levamisole (96.83%). Conclusion: The results show that the E. indica extract can reduce the fecal EPG counts of natu¬rally infected goats, thus creating a potential natural anthelmintic that can be developed further.

Objective: This study aims to determine the protein profile based on molecular weight (MW) and testosterone levels in seminal plasma (SP) that correlates to the semen quality of candidate Madura bulls. Material and Methods: A total of 10 male candidate madura bulls underwent semen evaluation (motility, viability, membrane plasma integrity (MPI), and sperm concentration). The centrifuge was run at 1,200 rpm (4°C) for 20 min to collect SP. SP testosterone levels were measured using an Enzyme-linked immunosorbent assay. The characterization of SP proteins in Madura bulls was done using 1D sodium dodecyl-sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. All parameters were analyzed using Pearson correlation analysis. Result: The results of the SDS-PAGE analysis found eight protein bands with the highest MW of 110 kDa and the lowest of 12 kDa. The mean and SD of SP testosterone levels were 20.58 ± 8.56 ng/ml, motility 59.32% ± 20.14%, viability 67.45% ± 20.22%, MPI 32.77% ± 16.52%, and sperm concentration 1,002.64 ± 429.33 106/mm3. Proteins with MWs of 110 and 91 kDa significantly correlated with MPI, and 110 kDa negatively correlated with sperm concentration (p < 0.05). Proteins with MWs of 73 and 36 kDa significantly correlated with SP testosterone levels, while proteins with MWs of 29 kDa significantly correlated with sperm viability (p < 0.05). Conclusion: The expressed protein fraction based on MW is closely related to the quality of semen, so it has the potential to be a biomarker of semen quality. Further research is needed to determine the specific proteins in certain fractions.

Objective: This research aims to identify the effect of various organic solvents such as n-hexane, ethyl acetate (EtOAc), and methanol (MeOH) on the antioxidant and antibacterial activities of Curcuma zedoaria extract, against three Gram-positive bacteria, namely Staphylococcus aureus, Bacillus subtilis, and Streptococcus pneumoniae, and three Gram-negative bacteria, namely Escherichia coli, Salmonella typhi, and Pseudomonas aeruginosa. Materials and Methods: As much as 1 kg of white turmeric rhizome (C. zedoaria) was extracted two times for 24 h using 3 l of MeOH before evaporating. The extract was then fractionated using n-hexane six times per 2 h, with each volume of 500 ml, and continued with the EtOAc fractionation. The MeOH fraction was added to 300 ml of water before adding 400 ml of EtOAc. Once the fractionation process was complete, all fractions were concentrated using a rotary evaporator. Results: The C. zedoria extract fractioned using MeOH produces alkaloids, phenolics, flavonoids, saponins, and coumarin compounds. The fractionation with EtOAc also produces alkaloids, phenolics, flavonoids, saponins, coumarin compounds, and triterpenoids. Meanwhile, fractionation with n-hexane only produces alkaloids and triterpenoid compounds. EtOAc and MeOH fractions had good activity in reducing free radicals produced by 2,2-diphenyl-1-picrylhydrazyl (DPPH), with an average IC50 value of 153.49 ± 2.66 and 185.77 ± 3.91 ppm, respectively. In contrast, the n-hexane fraction has weak antioxidant activity with an IC50 value of 837.92 ± 5.32 ppm. The n-hexane fraction has better activity compared to MeOH and EtOAc. The lowest concentration required was 2,500 ppm for all types of bacteria. Conclusion: Curcuma zedoaria extract produces alkaloids, phenolics, flavonoids, saponins, coumarins, and triterpenoids when fractionated with MeOH or EtOAc. Only alkaloids and triterpenoids are produced using n-hexane. EtOAc and MeOH fractions have good activity in reducing free radicals generated by DPPH, with an average IC50 value of 153.49 ± 2.66 and 185.77 ± 3.91 ppm, respectively. However, n-hexane has weak antioxidant activity, with an average IC50 value of 837.92 ± 5.32 ppm. All fractions have moderate antibacterial activity, but the extract of n-hexane from C. zedoary has better antibacterial activity compared to MeOH and EtOAc. The lowest concentration required is 2,500 ppm for all types of bacteria.

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: This work was conducted for the development of a 5-combi lateral flow immunochro¬matographic kit (LFK) for rapid and simultaneous identification of the common bacterial causes of bovine mastitis. The following pathogens are the identification targets of this kit: Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Streptococcus agalactiae, and Streptococcus pyogenes in milk samples from suspected bovine mastitis cases. The conventional microbiological identification of these agents is not only time-consuming and requires a fully equipped laboratory but also requires experienced personnel. Materials and Methods: Rabbit polyclonal antibodies (PAbs) specific to the antigenic components of the selected pathogens were prepared, and the pathogen-specific IgG was separated, purified, and conjugated with nanogold that was laid on the conjugate pad. Guinea pig PAbs specific to the microbial antigens of the selected pathogens were prepared, and their IgG content was separated, purified, and used as a capture antibody in the test (T) line on the nitrocellulose (NC) strips. Goat anti-rabbit IgG antibodies were used to capture the rabbit antibodies in the control (C) line of NC strips. The kit was held in a device comprising five strip-holding channels for the above-mentioned five bacterial species antigens. The developed LFK was evaluated, and its sensitivity and specificity were determined. Results: The developed kits were applied for the examination of bovine milk samples from suspected mastitis cases, and the average sensitivity, specificity, and accuracy of 5-combi LFK for the detection of the five selected bacterial species compared to bacteriological examination (gold standard test) were 93.90%, 80.83%, and 90.53%, respectively. The minimal microbial count that gave positive results using the developed LFK was 103 colony forming unit/ml. Treatment of the milk samples with an application buffer and its pre-incubation in trypticase soy broth for 6 h at 37°C before testing significantly increased the sensitivity of the prepared LFK. The developed kit proved simple and convenient, and the results could be obtained in less than 10 min. [J Adv Vet Anim Res 2023; 10(2.000): 292-300]

Objectives: The study aimed to account for baseline biometrical and histomorphometric testicu¬lar changes in Black Bengal goats during postnatal development. Materials and Methods: Black Bengal goats, divided into group I of VII; day 0; 1, 2 weeks; 1, 2, 4, and 6 months of age, respectively, were used in this study. Results: The biometrical and histomorphometric values of the testis varied significantly (p < 0.05) from postnatal 1–2 months. From day 0 to 2 months, seminiferous tubules, called sex cords, contained simply peripherally placed Sertoli cells and centrally placed gonocytes. Gonocytes, posi¬tioned in the center, moved centrifugally in the direction of the basement membrane of sex cords with the advancement of age, transformed into prespermatogonia, and were distributed among the Sertoli cells at the edge of sex cords that make up the basal cell layer in nearly all of the sem¬iniferous tubules by 2 months after birth. Initiation of spermatogenesis, i.e., stratification and lumination of seminiferous epithelium, took place in the 4th months. At 6 months, all types of spermatogenic cells had been identified. The onset of puberty, i.e., the establishment of sper¬matogenesis, was noticed to have been established at 6 months of postnatal age in Black Bengal goats, as shown by the spermatozoa that were adhered to the ad luminal border of the Sertoli cells and also in the tubular lumen. Conclusion: This research is the first to document the varying biometrical and histomorphometric measurements of the testis in Black Bengal goats from birth to puberty. [J Adv Vet Anim Res 2023; 10(2.000): 237-243]

Objective: This study was conducted to validate polymerase chain reaction (PCR) as a confirma¬tory diagnostic tool to find out the presence and frequency of Streptococcus agalactiae (S. aga¬lactiae) and Streptococcus dysgalactiae (S. dysgalactiae) in mastitic milk samples obtained from dairy cows in the southern region of Bangladesh. Materials and Methods: A total of 196 samples of bovine milk were collected from various dairy farms in the Chattogram metropolitan area of the southern part of Bangladesh. DNA extracted from isolates obtained by culturing California mastitis test (CMT)-positive mastitic milk samples (n = 146) on 5% sheep blood agar was used as a template for PCR. Two sets of specific primers based on the 16S rRNA gene were used to discriminate between S. agalactiae and S. dysgalactiae. Four PCR products were subjected to sequencing, followed by phylogenetic analysis. Results: The PCR analyses revealed that out of the 146 CMT-positive milk samples tested, 29 samples were positive for S. agalactiae (19.86%), while 26 samples were positive for S. dysgalac¬tiae (17.81%). Further sequence analysis of the corresponding PCR products and bioinformatics analysis verified the results. Conclusion: The study proves the efficiency of PCR as a useful diagnostic approach to determine the presence and prevalence of S. agalactiae and S. dysgalactiae in mastitic milk samples obtained from dairy cows. [J Adv Vet Anim Res 2023; 10(2.000): 178-184]