Bee venom with an antimicrobial effect is a powerful natural product. One of the most important areas where new antimicrobials are needed is in the prevention and control of multi-drug resistant pathogens. Today, antibacterial products used to treat multi-drug resistant pathogen infections in hospitals and healthcare facilities are insufficient to prevent colonisation and spread, and new products are needed. The aim of the study is to investigate the antibacterial effect of the bee venom (BV), a natural substance, on the species of Methicillin resistant Staphylococcus aureus, Vancomycin resistant Enterococcus faecalis, Carbapenem resistant Escherichia coli, Carbapenem resistant Klebsiella pneumoniae and Carbapenem resistant Acinetobacter baumannii. As a result of this study, it was found that MIC90 and MBC90 values ranged from 6.25 μg/mL – 12.5 μg/mL and numbers of bacteria decreased by 4–6 logs within 1–24 h for multi-drug resistant pathogens. In particular, Vancomycin resistant Enterococcus faecalis isolate decreased 6 log cfu/mL at 50 μg/mL and 100 μg/mL concentrations in the first hour. The effective bacterial inhibition rate of bee venom suggests that it could be a potential antibacterial agent for multi-drug resistant pathogens.Contribution: The treatment options of antibiotic-resistant pathogens are a major problem in both veterinary and human medicine fields. We have detected a high antibacterial effect against these agents in this bee venom study, which is a natural product. Apitherapy is a fashionable treatment method all over the world and is used in many areas of health. Bee venom is also a product that can be used as a drug or disinfectant raw material and can fill the natural product gap that can be used against resistant bacteria.

Bee venom with an antimicrobial effect is a powerful natural product. One of the most important areas where new antimicrobials are needed is in the prevention and control of multi-drug resistant pathogens. Today, antibacterial products used to treat multi-drug resistant pathogen infections in hospitals and healthcare facilities are insufficient to prevent colonisation and spread, and new products are needed. The aim of the study is to investigate the antibacterial effect of the bee venom (BV), a natural substance, on the species of Methicillin resistant Staphylococcus aureus, Vancomycin resistant Enterococcus faecalis, Carbapenem resistant Escherichia coli, Carbapenem resistant Klebsiella pneumoniae and Carbapenem resistant Acinetobacter baumannii. As a result of this study, it was found that MIC90 and MBC90 values ranged from 6.25 μg/mL – 12.5 μg/mL and numbers of bacteria decreased by 4–6 logs within 1–24 h for multi-drug resistant pathogens. In particular, Vancomycin resistant Enterococcus faecalis isolate decreased 6 log cfu/mL at 50 μg/mL and 100 μg/mL concentrations in the first hour. The effective bacterial inhibition rate of bee venom suggests that it could be a potential antibacterial agent for multi-drug resistant pathogens. Contribution: The treatment options of antibiotic-resistant pathogens are a major problem in both veterinary and human medicine fields. We have detected a high antibacterial effect against these agents in this bee venom study, which is a natural product. Apitherapy is a fashionable treatment method all over the world and is used in many areas of health. Bee venom is also a product that can be used as a drug or disinfectant raw material and can fill the natural product gap that can be used against resistant bacteria.

Objective: Certain advantages of donkeys are still not listed as for other equine species. Moreover, donkeys lack comprehensive scientific studies. The present study examines the histological architecture and histochemical characteristics of the esophagus in the Iraqi local breed donkey (Equus asinus). Materials and Methods: Eight esophagus samples were collected from a local breed donkey. Tissue specimens (~1 cm3 ) were collected from the cervical, thoracic, and abdominal regions of the esophagus and processed via routine histological technique. The tissue sections were stained with hematoxylin and eosin, Massons Trichrome, and combined Alcian blue (pH 2.5) plus PAS (AB-PAS). Results: The esophagus of the local breed donkey had folded mucosa wrapped by thin non-keratinized stratified squamous epithelium. The heights of epithelia of the cervical and thoracic regions of the esophagus were significantly higher than that of the abdominal regions. The lamina propria consisted of dense fibrous tissue that appeared thickest in the thoracic and abdominal regions of the esophagus. The muscularis mucosa disappears at the cervical region, while the thoracic and abdominal regions of the esophagus contain thick, scattered, interrupted bundles of smooth muscle fibers. Tunica submucosa was very thick at the thoracic and abdominal regions of the esophagus, composed of loose connective tissue filled with compound tubular mucoserous esophageal glands. Using a combined AB-PAS stain, mucous alveoli within the esophageal glands indicated strong acidic mucopolysaccharide. Tunica muscularis of the cervical and thoracic regions was built up by striated muscle fibers and turned into smooth type at the abdominal region of the esophagus. Conclusion: The esophagus of the local breed donkey shows considerable histological similarities with the other mammals that make this species reliable as an experimental model of digestive tissue. J. Adv. Vet. Anim. Res., 10(1): 14–20, March 2023 http://doi.org/10.5455/javar.2023.j647

Objective: Certain advantages of donkeys are still not listed as for other equine species. Moreover, donkeys lack comprehensive scientific studies. The present study examines the histological architecture and histochemical characteristics of the esophagus in the Iraqi local breed donkey (Equus asinus). Materials and Methods: Eight esophagus samples were collected from a local breed donkey. Tissue specimens (~1 cm³) were collected from the cervical, thoracic, and abdominal regions of the esoph¬agus and processed via routine histological technique. The tissue sections were stained with hema¬toxylin and eosin, Massons Trichrome, and combined Alcian blue (pH 2.5) plus PAS (AB-PAS). Results: The esophagus of the local breed donkey had folded mucosa wrapped by thin non-ke¬ratinized stratified squamous epithelium. The heights of epithelia of the cervical and thoracic regions of the esophagus were significantly higher than that of the abdominal regions. The lamina propria consisted of dense fibrous tissue that appeared thickest in the thoracic and abdominal regions of the esophagus. The muscularis mucosa disappears at the cervical region, while the thoracic and abdominal regions of the esophagus contain thick, scattered, interrupted bundles of smooth muscle fibers. Tunica submucosa was very thick at the thoracic and abdominal regions of the esophagus, composed of loose connective tissue filled with compound tubular mucose¬rous esophageal glands. Using a combined AB-PAS stain, mucous alveoli within the esophageal glands indicated strong acidic mucopolysaccharide. Tunica muscularis of the cervical and thoracic regions was built up by striated muscle fibers and turned into smooth type at the abdominal region of the esophagus. Conclusion: The esophagus of the local breed donkey shows considerable histological similarities with the other mammals that make this species reliable as an experimental model of digestive tissue. [J Adv Vet Anim Res 2023; 10(1.000): 14-20]

Objective: The research was conducted to calculate feed, calcium (Ca) intake, Ca requirements, and egg production for Arabic hens during the early egg-laying period. Materials and Methods: A total of 135 30-week-old Silver female Arabic pullets were randomly allocated to one of three treatments with five replicate cages with a semi-scavenging system of nine pullets per cage in a completely randomized design and allowed to choose Ca from limestone and oyster shells. As a control (T1), pullets were only given a complete feed with Ca and phosphorus percentages according to Hy-line International in 2018. Other treatment feeds were control feed without limestone fed with a combination of limestone (T2) or oyster shell (T3) separately. Results: The treatments had no effect (p > 0.05) on feed, grit and Ca intake (gm/bird/week), egg weight (gm), egg mass, egg production (%), and feed efficiency, but had an effect (p < 0.05) on Ca concentration (%). Ca concentration was the same at T1 and T3, and both were higher than at T2. Conclusion: The female Arabic chickens could fulfill Ca requirements by selecting from different sources of Ca. Limestone is better than the oyster shell as a source of Ca. The Ca requirement for Arabic hensin the early laying period based on the Ca concentration of the feed intake is sufficient at around 3.64% since producing the same number of egg production and heavier egg weight compared to a higher Ca level. J. Adv. Vet. Anim. Res., 10(1): 30–41, March 2023 http://doi.org/10.5455/javar.2023.j649

Objective: The research was conducted to calculate feed, calcium (Ca) intake, Ca requirements, and egg production for Arabic hens during the early egg-laying period. Materials and Methods: A total of 135 30-week-old Silver female Arabic pullets were randomly allocated to one of three treatments with five replicate cages with a semi-scavenging system of nine pullets per cage in a completely randomized design and allowed to choose Ca from limestone and oyster shells. As a control (T1), pullets were only given a complete feed with Ca and phosphorus percentages according to Hy-line International in 2018. Other treatment feeds were control feed without limestone fed with a combination of limestone (T2) or oyster shell (T3) separately. Results: The treatments had no effect (p > 0.05) on feed, grit and Ca intake (gm/bird/week), egg weight (gm), egg mass, egg production (%), and feed efficiency, but had an effect (p < 0.05) on Ca concentration (%). Ca concentration was the same at T1 and T3, and both were higher than at T2. Conclusion: The female Arabic chickens could fulfill Ca requirements by selecting from different sources of Ca. Limestone is better than the oyster shell as a source of Ca. The Ca requirement for Arabic hens in the early laying period based on the Ca concentration of the feed intake is sufficient at around 3.64% since producing the same number of egg production and heavier egg weight compared to a higher Ca level. [J Adv Vet Anim Res 2023; 10(1.000): 30-41]

Objective: This study was conducted to validate polymerase chain reaction (PCR) as a confirmatory diagnostic tool to find out the presence and frequency of Streptococcus agalactiae (S. agalactiae) 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. dysgalactiae (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., 10(2): 178–184, June 2023 http://doi.org/10.5455/javar.2023.j667

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]

Objective: Arcobacter butzleri, Listeria monocytogenes, Staphylococcus aureus, and Campylobacter jejuni are significant foodborne pathogens regarding the consumption of raw poultry meat. An existing survey was conducted to assess the occurrence of S. aureus, C. jejuni, A. butzleri, and L. monocytogenes in raw poultry meat samples. Materials and Methods: Ninety-four raw ostrich, turkey, chicken, and quail meat samples were collected and subjected to culture-based analysis. Staphylococcus aureus, C. jejuni, A. butzleri, and L. monocytogenes isolates were confirmed by standard biochemical techniques. Results: The occurrence of A. butzleri, C. jejuni, L. monocytogenes, and S. aureus in poultry meat samples was 11.45%, 17.70%, 1.04%, and 16.66%, respectively. L. monocytogenes was absent in chicken, turkey, and ostrich meat samples. Only one quail meat (4.16%) was positive for L. monocytogenes. The uppermost contamination rate with A. butzleri, C. jejuni, and S. aureus was found in chicken (25%), turkey (25%), and turkey (25%) meat samples, respectively. The concurrent occurrence of A. butzleri + C. jejuni + S. aureus bacteria amid the examined poultry meat samples was 2.08%. Conclusion: This is an initial report of A. butzleri, S. aureus, C. jejuni, and L. monocytogenes in poultry meat samples. Adequate cooking of poultry meat can diminish foodborne diseases due to A. butzleri, S. aureus, L. monocytogenes, and C. jejuni bacteria, and these species may constitute a public health problem.  J. Adv. Vet. Anim. Res., 10(2): 205–210, June 2023 http://doi.org/10.5455/javar.2023.j670

Objective: Arcobacter butzleri, Listeria monocytogenes, Staphylococcus aureus, and Campylobacter jejuni are significant foodborne pathogens regarding the consumption of raw poultry meat. An existing survey was conducted to assess the occurrence of S. aureus, C. jejuni, A. butzleri, and L. monocytogenes in raw poultry meat samples. Materials and Methods: Ninety-four raw ostrich, turkey, chicken, and quail meat samples were collected and subjected to culture-based analysis. Staphylococcus aureus, C. jejuni, A. butzleri, and L. monocytogenes isolates were confirmed by standard biochemical techniques. Results: The occurrence of A. butzleri, C. jejuni, L. monocytogenes, and S. aureus in poultry meat samples was 11.45%, 17.70%, 1.04%, and 16.66%, respectively. L. monocytogenes was absent in chicken, turkey, and ostrich meat samples. Only one quail meat (4.16%) was positive for L. monocytogenes. The uppermost contamination rate with A. butzleri, C. jejuni, and S. aureus was found in chicken (25%), turkey (25%), and turkey (25%) meat samples, respectively. The concur¬rent occurrence of A. butzleri + C. jejuni + S. aureus bacteria amid the examined poultry meat samples was 2.08%. Conclusion: This is an initial report of A. butzleri, S. aureus, C. jejuni, and L. monocytogenes in poultry meat samples. Adequate cooking of poultry meat can diminish foodborne diseases due to A. butzleri, S. aureus, L. monocytogenes, and C. jejuni bacteria, and these species may constitute a public health problem. [J Adv Vet Anim Res 2023; 10(2.000): 205-210]

Objective: The objective of the present study was to evaluate the effects of using dried chaya leaf meal (DCLM) as a protein feed in a diet on growth performance, blood profiles, and carcass quality in a native Thai chicken. Materials and Methods: Eighty 14-day-old Pradu Hangdum chicks were divided into four groups with four replicates each: control (without DCLM), 10%, 20%, and 30% DCLM inclusion in mash feed. Growth performance was recorded weekly until 98 days of age. Blood profile, carcass quality, and visceral organ weight were measured at 98 days of age. Results: The 10%–30% DCLM inclusion in the diet did not affect feed intake or feed efficiency; however, the body weight gain of chicks decreased linearly with the increase in DCLM inclusion. The heterophils, eosinophils, and monocytes were linearly increased with the increasing DCLM levels in the groups. The serum blood chemistry did not differ among the groups, while the AST in 10% and 20% DCLM was lower than in the control. Increasing the level of DCLM in the chicken diet did not affect carcass quality. Conclusion: The DCLM can be used as a feed ingredient in Thai native chicken feed up to 20%. J. Adv. Vet. Anim. Res., 10(1): 51–56, March 2023 http://doi.org/10.5455/javar.2023.j651

Objective: The objective of the present study was to evaluate the effects of using dried chaya leaf meal (DCLM) as a protein feed in a diet on growth performance, blood profiles, and carcass quality in a native Thai chicken. Materials and Methods: Eighty 14-day-old Pradu Hangdum chicks were divided into four groups with four replicates each: control (without DCLM), 10%, 20%, and 30% DCLM inclusion in mash feed. Growth performance was recorded weekly until 98 days of age. Blood profile, carcass quality, and visceral organ weight were measured at 98 days of age. Results: The 10%–30% DCLM inclusion in the diet did not affect feed intake or feed efficiency; however, the body weight gain of chicks decreased linearly with the increase in DCLM inclusion. The heterophils, eosinophils, and monocytes were linearly increased with the increasing DCLM levels in the groups. The serum blood chemistry did not differ among the groups, while the AST in 10% and 20% DCLM was lower than in the control. Increasing the level of DCLM in the chicken diet did not affect carcass quality. Conclusion: The DCLM can be used as a feed ingredient in Thai native chicken feed up to 20%. [J Adv Vet Anim Res 2023; 10(1.000): 51-56]

Objective: This study aimed to establish and focus on the implications of precise energy and protein levels on reproductive performance. This study based on field facts showed that excessive feed intake, especially protein intake, to increase productivity will lead to some decreased reproductive efficiency through disruption of reproductive hormonal compound biosynthesis or increasing blood urea nitrogen (BUN), which has rarely been studied. Materials and Methods: Healthy and mature Saanen crossbred goats raised by traditional farmers (2–4 years old; ± 45 kg) were divided into three experimental groups [T0 = basal feed according to the feed provided by farmers, T1 = protein and energy balanced feed according to National Research Council (NRC) (±5%), T2 = feed >20% excess protein, and excess energy from NRC] (n = 75). The factorial method was used in the experimental design with a post-hoc least significant difference test. Results: The data showed a significant difference (p < 0.05) in the biochemical or hormonal parameters between the control and the treatment groups. Also, T1 showed leads in any blood profile and reproductive hormone parameters such as total protein (79.6 ± 5.3 gm/dl), cholesterol (3.26 ± 0.47 mmol/l), progesterone (1.79 ± 2.21 ng/ml), and estrogen (47.85 ± 5.51 pg/ml), as well as the lowest levels of BUN (11.6 ± 1.59 mmol/l) and cortisol (25.07 ± 14.85 ng/ml) levels. T1 had the highest reproductive potential after treatment. Conclusion: The adverse effects of high and low protein consumption on reproductive hormones seem to be related to the blood profile and ovarian function, as hormone concentrations change significantly and lead to reproductive impairment. The data showed that balanced nutrient levels (5%/T1) resulted in excellent blood and hormone parameters.  J. Adv. Vet. Anim. Res., 10(2): 257–268, June 2023 http://doi.org/10.5455/javar.2023.j677

Objective: This study aimed to establish and focus on the implications of precise energy and protein levels on reproductive performance. This study based on field facts showed that exces¬sive feed intake, especially protein intake, to increase productivity will lead to some decreased reproductive efficiency through disruption of reproductive hormonal compound biosynthesis or increasing blood urea nitrogen (BUN), which has rarely been studied. Materials and Methods: Healthy and mature Saanen crossbred goats raised by traditional farm¬ers (2–4 years old; ± 45 kg) were divided into three experimental groups [T0 = basal feed according to the feed provided by farmers, T1 = protein and energy balanced feed according to National Research Council (NRC) (±5%), T2 = feed >20% excess protein, and excess energy from NRC] (n = 75). The factorial method was used in the experimental design with a post-hoc least significant difference test. Results: The data showed a significant difference (p < 0.05) in the biochemical or hormonal parameters between the control and the treatment groups. Also, T1 showed leads in any blood profile and reproductive hormone parameters such as total protein (79.6 ± 5.3 gm/dl), cholesterol (3.26 ± 0.47 mmol/l), progesterone (1.79 ± 2.21 ng/ml), and estrogen (47.85 ± 5.51 pg/ml), as well as the lowest levels of BUN (11.6 ± 1.59 mmol/l) and cortisol (25.07 ± 14.85 ng/ml) levels. T1 had the highest reproductive potential after treatment. Conclusion: The adverse effects of high and low protein consumption on reproductive hormones seem to be related to the blood profile and ovarian function, as hormone concentrations change significantly and lead to reproductive impairment. The data showed that balanced nutrient levels (5%/T1) resulted in excellent blood and hormone parameters. [J Adv Vet Anim Res 2023; 10(2.000): 257-268]

Objective: The present study analyzed the seminal plasma proteome and possible relationships between proteins and semen quality in azoospermic and normal Simmental bulls. Materials and Methods: Fresh semen plasma samples from the Lembang Artificial Insemination Center were used for this study, including one bull (76´ ejaculate) with very poor semen quality/ azoospermia (poor fresh semen/infertile; PFS) and three bulls with normal semen quality (normal fresh semen; NFS) for proteomic analysis using a pooled system (NFS-Stud) (60´ ejaculate). The only males obtained with very low quality or azoospermia (PFS) had sperm motility of <10% (one head). Bulls with azoospermic conditions produce fresh semen without sperm or with very little sperm concentration. A total of 109 proteins were identified in the seminal plasma of Simmental bulls analyzed using liquid chromatography-mass spectrometry. Bioinformatics analysis was used to explore total protein, expression, function, and protein mechanism in the seminal plasma of Simmental bulls. Results: The results showed that the seminal plasma proteins expressed in NFS bulls include ELSPBP1, SIL1, HSPA13, angiotensin-1 covering enzyme, and CRISP1. On the other hand, B2M, C3, CFB, venin-2, and cathepsin S contribute significantly to PFS. The NFS bull proteins play important roles in sperm capacitation, protein transport, sperm motility, spermatogenesis, immune tolerance, and fertilization, while the PFS proteins perform apoptotic and antigen pathway functions. Conclusion: There is an interaction between proteins in the seminal plasma of males with poor semen quality (PFS) and cases of infertility (azoospermia) that cause a decrease in sperm quality in PFS bulls. J. Adv. Vet. Anim. Res., 10(3): 370–377, September 2023 http://doi.org/10.5455/javar.2023.j689

Objective: The present study analyzed the seminal plasma proteome and possible relationships between proteins and semen quality in azoospermic and normal Simmental bulls. Materials and Methods: Fresh semen plasma samples from the Lembang Artificial Insemination Center were used for this study, including one bull (76´ ejaculate) with very poor semen quality/ azoospermia (poor fresh semen/infertile; PFS) and three bulls with normal semen quality (normal fresh semen; NFS) for proteomic analysis using a pooled system (NFS-Stud) (60´ ejaculate). The only males obtained with very low quality or azoospermia (PFS) had sperm motility of