Objective: The present study aims to investigate molecular confirmation for Cryptosporidium species in fish handlers in Baghdad City, central Iraq. Materials and Methods: Sixty stool samples were collected between early November 2023 and late April 2024. All samples were examined phenotypically using a modified Ziehl-Neelsen stain and genotypically (nested polymerase chain reaction technique) based on a partial sequence of 18S rRNA genes with sequencing and phylogenetic tree analysis. Results: The total molecular results identified Cryptosporidium parvum with an infection rate of 45% (27/60). A higher infection rate of 51.9% (14/27) was found in the age group between 15 and 35 years, and male handlers recorded a lower infection rate (45%) than females (41.6%). April had a higher elevation in the infection rate of 60% (6/10) than other months. Conclusion: The C. parvum was the only species found in fish handlers, and these local isolates have higher similarity with other isolates of China and Iran. J. Adv. Vet. Anim. Res., 12(1): 1–7, March 2025 http://doi.org/10.5455/javar.2025.l866

Objective: The present study aims to investigate molecular confirmation for Cryptosporidium species in fish handlers in Baghdad City, central Iraq. Materials and Methods: Sixty stool samples were collected between early November 2023 and late April 2024. All samples were examined phenotypically using a modified Ziehl-Neelsen stain and genotypically (nested polymerase chain reaction technique) based on a partial sequence of 18S rRNA genes with sequencing and phylogenetic tree analysis. Results: The total molecular results identified Cryptosporidium parvum with an infection rate of 45% (27/60). A higher infection rate of 51.9% (14/27) was found in the age group between 15 and 35 years, and male handlers recorded a lower infection rate (45%) than females (41.6%). April had a higher elevation in the infection rate of 60% (6/10) than other months. Conclusion: The C. parvum was the only species found in fish handlers, and these local isolates have higher similarity with other isolates of China and Iran. [J Adv Vet Anim Res 2025; 12(1.000): 1-7]

Objective: This study aims to explore mutation based on single nucleotide polymorphism (SNP) in the Cholecystokinin B receptor (CCKBR) gene of Pelung chickens. Materials and Methods: We collected DNA samples from 48 Pelung roosters that had won the crowing competition. The CCKBR target encompasses exon 3, intron 3, exon 4, and a part of intron 4, a long 601 bp. This target was replicated using PCR with specific primers that were designed by Primer-BLAST from NCBI. We generated the nucleotide sequence from the PCR product’s sequencing results. The SNP analysis was done by BioEdit and MEGA. Genotyping and haplotyping were done based on nonsynonymous single nucleotide polymorphisms (SNPs) on exons 3 and 4. We calculated allele and genotype frequency, heterozygosity, and Hardy-Weinberg Equilibrium (HWE) using POPGENE 32 programs. Results: This study found three nonsynonymous single nucleotide polymorphisms. The nsSNP in exon 3 altersthe coding for the 210th amino acid from serine to asparagine (g.1290 G > A/S210N), while the SNPs in exon 4 alter the coding for the 232nd amino acid from valine to phenylalanine (g.1423G > T/V232F) and the 243rd amino acid that changes the amino acid valine to glycine (g.1457T > G/V243G). The frequency of the mutated alleles is lower than the unmutated alleles. However, the mutation at position g.1457T > G/V243G produces a higher frequency than the unmutated allele. The allele and genotype frequency were not in HWE. It was caused by intensive selection in Pelung chickens, especially for growing capacity. Conclusion: Nonsynonymous mutation on CCKBR may cause variations in the crowing and other traits such as the growth of Pelung chickens. Further studies are needed to explore the CCKBR gene, including the relationship of the gene with the vigor and/or stress level of Pelung chickens. J. Adv. Vet. Anim. Res., 12(1): 141–148, March 2025 http://doi.org/10.5455/javar.2025.l881

Objective: This study aims to explore mutation based on single nucleotide polymorphism (SNP) in the Cholecystokinin B receptor (CCKBR) gene of Pelung chickens. Materials and Methods: We collected DNA samples from 48 Pelung roosters that had won the crowing competition. The CCKBR target encompasses exon 3, intron 3, exon 4, and a part of intron 4, a long 601 bp. This target was replicated using PCR with specific primers that were designed by Primer-BLAST from NCBI. We generated the nucleotide sequence from the PCR product's sequencing results. The SNP analysis was done by BioEdit and MEGA. Genotyping and haplotyp¬ing were done based on nonsynonymous single nucleotide polymorphisms (SNPs) on exons 3 and 4. We calculated allele and genotype frequency, heterozygosity, and Hardy-Weinberg Equilibrium (HWE) using POPGENE 32 programs. Results: This study found three nonsynonymous single nucleotide polymorphisms. The nsSNP in exon 3 alters the coding for the 210th amino acid from serine to asparagine (g.1290 G > A/S210N), while the SNPs in exon 4 alter the coding for the 232nd amino acid from valine to phenylalanine (g.1423G > T/V232F) and the 243rd amino acid that changes the amino acid valine to glycine (g.1457T > G/V243G). The frequency of the mutated alleles is lower than the unmutated alleles. However, the mutation at position g.1457T > G/V243G produces a higher frequency than the unmutated allele. The allele and genotype frequency were not in HWE. It was caused by intensive selection in Pelung chickens, especially for growing capacity. Conclusion: Nonsynonymous mutation on CCKBR may cause variations in the crowing and other traits such as the growth of Pelung chickens. Further studies are needed to explore the CCKBR gene, including the relationship of the gene with the vigor and/or stress level of Pelung chickens. [J Adv Vet Anim Res 2025; 12(1.000): 141-148]

Objective: This research was designed to explore the potential of mycotoxin binders derived from zeolite and bioherbal formulations as natural feed additives to enhance growth performance and intestinal characteristics in broilers. Materials and Methods: The study utilized 320 Lohmann MB 202 broilers, sourced from PT. Japfa Comfeed Indonesia, commencing from day 1 and extending over a period of 35 days. The methodological framework employed a completely eandomized design, incorporating two factors. The primary factor analyzed was the type of feed additive, designated as Zeolite (A1) and Bioherbal (A2). The secondary factor considered was the level of mycotoxin binder inclusion in the feed, set at four increments: 0% (T1), 0.2% (T2), 0.4% (T3), and 0.6% (T4), resulting in a total of eight treatment combinations, each replicated four times. The observational metrics focused on production performance and specific intestinal characteristics of the broilers. Results: The findings indicated that while the interaction between feed type and the level of additive use did not significantly influence feed consumption, weight gain, feed conversion ratio, or villi length (p > 0.05), there was a notable impact on the villi surface area (p < 0.05) and a pronounced effect on villi count and crypt depth (p < 0.01). Conclusion: The study concluded that mycotoxin binders containing zeolite effectively reduce mycotoxin levels in feed, whereas bioherbal additives significantly improve intestinal health. Thus, a 0.6% inclusion level of these additives is recommended as a viable alternative to antibiotics in broiler chicken diets. J. Adv. Vet. Anim. Res., 12(1): 149–156, March 2025 http://doi.org/10.5455/javar.2025.l882

Objective: This research was designed to explore the potential of mycotoxin binders derived from zeolite and bioherbal formulations as natural feed additives to enhance growth performance and intestinal characteristics in broilers. Materials and Methods: The study utilized 320 Lohmann MB 202 broilers, sourced from PT. Japfa Comfeed Indonesia, commencing from day 1 and extending over a period of 35 days. The methodological framework employed a completely randomized design, incorporating two factors. The primary factor analyzed was the type of feed additive, designated as Zeolite (A1) and Bioherbal (A2). The secondary factor considered was the level of mycotoxin binder inclusion in the feed, set at four increments: 0% (T1), 0.2% (T2), 0.4% (T3), and 0.6% (T4), resulting in a total of eight treat¬ment combinations, each replicated four times. The observational metrics focused on production performance and specific intestinal characteristics of the broilers. Results: The findings indicated that while the interaction between feed type and the level of additive use did not significantly influence feed consumption, weight gain, feed conversion ratio, or villi length (p > 0.05), there was a notable impact on the villi surface area (p < 0.05) and a pronounced effect on villi count and crypt depth (p < 0.01). Conclusion: The study concluded that mycotoxin binders containing zeolite effectively reduce mycotoxin levels in feed, whereas bioherbal additives significantly improve intestinal health. Thus, a 0.6% inclusion level of these additives is recommended as a viable alternative to antibiotics in broiler chicken diets. [J Adv Vet Anim Res 2025; 12(1.000): 149-156]

Objective: This research aims to isolate, screen, and identify some candidates for endophytic fungus-producing cellulase and cyanidase. Materials and Methods: Fungi were isolated from cassava leaves that had undergone surface sterilization. The fungal isolates were qualitatively tested for their ability to produce cellulase and cyanidase enzymes by adding carboxy methyl cellulose (CMC) and KCN to the media. Enzyme production was indicated by the formation of clear zones around the growing colonies. Isolates that tested positive for cellulase and cyanidase production underwent further quantitative screening to measure enzyme activity using a spectrophotometer at wavelengths of 540 nm and 400 nm, respectively. The isolates showing the highest cellulase and cyanidase activity were identified through 18S rRNA analysis using the Sanger DNA sequencing method. Results: The research obtained six pure isolates of endophytic fungus, namely Y1; Y2; Y3; Y4; Y5; and Y6. Four isolates had the ability to degrade CMC with a clear zone between 0.1 until 0.5 mm, and three isolates had the ability for KCN degrade. The highest activity for cellulase and cyanidase degrading enzymes was produced by isolate Y2. After molecular identification using 18S rRNA, isolate Y2 had 98.82% similarity to Phomopsis sp. 32PG/F. Conclusion: Six isolates of endophytic fungi were obtained, Y1; Y2; Y3; Y4; Y5; and Y6. Four isolate the ability of to degrade CMC and three isolate the ability for KCN degrade. Isolate Y2 is the isolate with the best activity for cellulase and cyanidase degrading enzymes, namely 2.99 U/ml and 2.19 U/ml. After molecular identification using 18S rRNA, isolate Y2 had 98.82% similarity to Phomopsis sp. 32PG/F. J. Adv. Vet. Anim. Res., 12(1): 169–178, March 2025 http://doi.org/10.5455/javar.2025.l884

Objective: This research aims to isolate, screen, and identify some candidates for endophytic fungus-producing cellulase and cyanidase. Materials and Methods: Fungi were isolated from cassava leaves that had undergone surface sterilization. The fungal isolates were qualitatively tested for their ability to produce cellulase and cyanidase enzymes by adding carboxy methyl cellulose (CMC) and KCN to the media. Enzyme production was indicated by the formation of clear zones around the growing colonies. Isolates that tested positive for cellulase and cyanidase production underwent further quantitative screening to measure enzyme activity using a spectrophotometer at wavelengths of 540 nm and 400 nm, respectively. The isolates showing the highest cellulase and cyanidase activity were identified through 18S rRNA analysis using the Sanger DNA sequencing method. Results: The research obtained six pure isolates of endophytic fungus, namely Y1; Y2; Y3; Y4; Y5; and Y6. Four isolates had the ability to degrade CMC with a clear zone between 0.1 until 0.5 mm, and three isolates had the ability for KCN degrade. The highest activity for cellulase and cyanidase degrading enzymes was produced by isolate Y2. After molecular identification using 18S rRNA, isolate Y2 had 98.82% similarity to Phomopsis sp. 32PG/F. Conclusion: Six isolates of endophytic fungi were obtained, Y1; Y2; Y3; Y4; Y5; and Y6. Four iso¬late the ability of to degrade CMC and three isolate the ability for KCN degrade. Isolate Y2 is the isolate with the best activity for cellulase and cyanidase degrading enzymes, namely 2.99 U/ml and 2.19 U/ml. After molecular identification using 18S rRNA, isolate Y2 had 98.82% similarity to Phomopsis sp. 32PG/F. [J Adv Vet Anim Res 2025; 12(1.000): 169-178]

Objectives: This study investigates the effects of 2 mT static magnetic field (SMF) exposure for 1 h on the expression of Stim1 and Itpr3 genes in hepatic cells of obese mice. By examining these critical regulators of calcium (Ca2+) signaling and cellular metabolism, the research aims to elucidate the role of SMF in modulating molecular pathways essential for Ca2+ homeostasis and metabolic regulation in the context of obesity. Materials and Methods: Liver samples were obtained from C57BL/6J mice and preserved in RNALater. The samples were divided into two main groups: the control group, which received a standard diet, and the obese group, which was exposed to a high-fat diet. Furthermore, the obese group was stratified based on the duration of SMF exposure, including intervals of 0, 2, 7, 14, and 21 days (1 h per day with an intensity of Bmax = 2 mT). Statistical tests were conducted with a significance level of p < 0.05. Results: The research findings highlighted a noteworthy increase in the relative expression of Stim1 and Itpr3 among obese mice exposed to SMF for 7 days (obe7) and those exposed for 14 days (obe14) in comparison to the obese group without SMF exposure. Both the obe7 and obe14 groups exhibited no significant difference in relative Stim1 expression when compared to the nonobese group. However, in terms of Itpr3 expression, the obe14 group did not show a significant difference from the non-obese mouse group. The results of the correlation analysis unveiled a substantial and robust correlation between the relative expression of Stim1 and Itpm3 (r = 0.627, p < 0.001). Conclusion: These findings suggest a potential link between SMF exposure, the expression of Ca2+ regulatory genes, and the intricate pathways involved in obesity-related molecular responses. J. Adv. Vet. Anim. Res., 12(1): 231–237, March 2025 http://doi.org/10.5455/javar.2025.l890

Objectives: This study investigates the effects of 2 mT static magnetic field (SMF) exposure for 1 h on the expression of Stim1 and Itpr3 genes in hepatic cells of obese mice. By examining these critical regulators of calcium (Ca2+) signaling and cellular metabolism, the research aims to elucidate the role of SMF in modulating molecular pathways essential for Ca2+ homeostasis and metabolic regulation in the context of obesity. Materials and Methods: Liver samples were obtained from C57BL/6J mice and preserved in RNALater. The samples were divided into two main groups: the control group, which received a standard diet, and the obese group, which was exposed to a high-fat diet. Furthermore, the obese group was stratified based on the duration of SMF exposure, including intervals of 0, 2, 7, 14, and 21 days (1 h per day with an intensity of Bmax = 2 mT). Statistical tests were conducted with a significance level of p < 0.05. Results: The research findings highlighted a noteworthy increase in the relative expression of Stim1 and Itpr3 among obese mice exposed to SMF for 7 days (obe7) and those exposed for 14 days (obe14) in comparison to the obese group without SMF exposure. Both the obe7 and obe14 groups exhibited no significant difference in relative Stim1 expression when compared to the non-obese group. However, in terms of Itpr3 expression, the obe14 group did not show a significant difference from the non-obese mouse group. The results of the correlation analysis unveiled a substantial and robust correlation between the relative expression of Stim1 and Itpr3 (r = 0.627, p < 0.001). Conclusion: These findings suggest a potential link between SMF exposure, the expression of Ca2+ regulatory genes, and the intricate pathways involved in obesity-related molecular responses. [J Adv Vet Anim Res 2025; 12(1.000): 231-237]

Objective: The objective of this investigation was to identify and detect the reassortant infectious bursal disease virus (IBDV) strain from broiler farms suspected of being infected. Materials and Methods: The broiler yielded 72 samples, including the spleen and bursa of Fabricius. The tissues underwent histological examination before being used in a typical PCR molecular investigation. Results: The strain was subsequently termed IBDV ASPVB. The IBDV ASPVB strain in Iraq has been identified as a novel reassortant strain based on the results of PCR, sequencing, and phylogenetic analysis of partial segments A and B. Segment A of this strain is derived from the highly pathogenic IBDV strain. In contrast, segment B is derived from other field reassortant strains. Infection with this strain might result in minor clinical symptoms but substantial damage to lymphoid organs, leading to compromised immunological responses. Conclusion: As a result of ongoing evolution, this study demonstrates that IBDV in Iraq exhibits a wide range of histological, genetic, and phenotypic variation; to our knowledge, this paper represents the first report of reassortant IBDV in Iraq.  J. Adv. Vet. Anim. Res., 12(2): 385–395, June 2025 http://doi.org/10.5455/javar.2025.l906

Objective: The objective of this investigation was to identify and detect the reassortant infectious bursal disease virus (IBDV) strain from broiler farms suspected of being infected. Materials and Methods: The broiler yielded 72 samples, including the spleen and bursa of Fabricius. The tissues underwent histological examination before being used in a typical PCR molecular investigation. Results: The strain was subsequently termed IBDV ASPVB. The IBDV ASPVB strain in Iraq has been identified as a novel reassortant strain based on the results of PCR, sequencing, and phylogenetic analysis of partial segments A and B. Segment A of this strain is derived from the highly pathogenic IBDV strain. In contrast, segment B is derived from other field reassortant strains. Infection with this strain might result in minor clinical symptoms but substantial damage to lymphoid organs, leading to compromised immunological responses. Conclusion: As a result of ongoing evolution, this study demonstrates that IBDV in Iraq exhibits a wide range of histological, genetic, and phenotypic variation; to our knowledge, this paper represents the first report of reassortant IBDV in Iraq. [J Adv Vet Anim Res 2025; 12(2.000): 385-395]

Objective: Antibiotic resistance is a worldwide health challenge. Close interaction with dogs carrying antibiotic-resistant zoonotic agents poses a risk to human health. The present study aimed to characterize antibiotic-resistant Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) isolated from dogs in Southern Benin. Materials and methods: A total of 336 swabs (112 buccal, 112 nasal, and 112 rectal) from 112 dogs living in the communes of Abomey-Calavi and Cotonou were analyzed for E. coli and S. aureus presence. Bacterial isolates were tested for antibiotic (penicillins, tetracyclines, aminoglycosides, cephalosporins, sulfonamides, and macrolides) susceptibility using the disc diffusion method, and antibiotic-resistant strains were characterized by the polymerase chain reaction method. Results: A 41.07% and 20.53% of dogs harbored E. coli and S. aureus, respectively. Escherichia coli and S. aureus isolates showed resistance to penicillin (100% and 81.48%), tetracycline (44.64% and 59.26%), and other antimicrobials tested. Escherichia coli isolates harbored resistance genes blaTEM (63.46%), tetA (62.50%), and strA-strB (55.56%). tetK (100%), tetM (100%), and blaZ (82.61%) were present in S. aureus isolates. Escherichia coli strains harbored virulence genes fimH (61.54%), kpsMTII (26.92%), fyuA (19.23%), and eae (1.92%), whereas 20.83% of S. aureus strains harbored pvl and fnbA. Conclusion: The results of the current study reveal the urgent need for stricter controls on antibiotic use. Implementing guidelines, responsible prescribing, and increasing public awareness are crucial steps to address this problem. J. Adv. Vet. Anim. Res., 12(2): 396–419, June 2025 http://doi.org/10.5455/javar.2025.l907

Objective: Antibiotic resistance is a worldwide health challenge. Close interaction with dogs carrying antibiotic-resistant zoonotic agents poses a risk to human health. The present study aimed to characterize antibiotic-resistant Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) isolated from dogs in Southern Benin. Materials and methods: A total of 336 swabs (112 buccal, 112 nasal, and 112 rectal) from 112 dogs living in the communes of Abomey-Calavi and Cotonou were analyzed for E. coli and S. aureus presence. Bacterial isolates were tested for antibiotic (penicillins, tetracyclines, aminoglycosides, cephalosporins, sulfonamides, and macrolides) susceptibility using the disc diffusion method, and antibiotic-resistant strains were characterized by the polymerase chain reaction method. Results: A 41.07% and 20.53% of dogs harbored E. coli and S. aureus, respectively. Escherichia coli and S. aureus isolates showed resistance to penicillin (100% and 81.48%), tetracycline (44.64% and 59.26%), and other antimicrobials tested. Escherichia coli isolates harbored resistance genes blaTEM (63.46%), tetA (62.50%), and strA-strB (55.56%). tetK (100%), tetM (100%), and blaZ (82.61%) were present in S. aureus isolates. Escherichia coli strains harbored virulence genes fimH (61.54%), kpsMTII (26.92%), fyuA (19.23%), and eae (1.92%), whereas 20.83% of S. aureus strains harbored pvl and fnbA. Conclusion: The results of the current study reveal the urgent need for stricter controls on antibiotic use. Implementing guidelines, responsible prescribing, and increasing public awareness are crucial steps to address this problem. [J Adv Vet Anim Res 2025; 12(2.000): 396-419]

Objective: Our study is conducted to identify serotypes, antibiotic resistance, heavy metal resistance, and virulent genes in Escherichia coli isolated from goats raised in small-scale farms in some provinces of the Mekong Delta, Vietnam. Material and Methods: A total of 203 E. coli isolates from goat feces were examined by PCR for serotypes (O8, O9, O25, O26, O45, O103, O146, O157, and O159), eight antibiotic-resistance genes, four heavy-metal-resistance genes, and four pathogenic genes. Results: By PCR, 20.20% of E. coli isolates belonging to serotypes O8 (6.40%), O45 (13.30%), and O159 (0.49%) were identified. Antibiotic-resistance genes were recorded at high rates in E. coli isolates, especially genes blaampC (98.52%), tetA (50.74%), sulII (34.48%), qnrA (20.69%), and aadA1 (20.69%). Moreover, 55.67% of these E. coli isolates harbored multiple antibiotic-resistance genes. Among heavy-metal-resistance genes, the gene czcD encoding for resistance to cobalt, zinc, and cadmium was the most prevalent (59.11%). In addition, the most frequent virulent gene was stx1 (15.27%), followed by gene stx2 (6.90%), eae, and hlyA (1.48%). Conclusion: These results revealed that goats were a natural reservoir of pathogenic E. coli serotypes, which could cause severe diseases in animals and humans. Moreover, these E. coli isolates showed a high ability to resist diverse antibiotics. Thus, managing the prevalence of pathogenic E. coli is essential for protecting public health in the Mekong Delta. J. Adv. Vet. Anim. Res., 12(2): 420–426, June 2025 http://doi.org/10.5455/javar.2025.l908

Objective: Our study is conducted to identify serotypes, antibiotic resistance, heavy metal resistance, and virulent genes in Escherichia coli isolated from goats raised in small-scale farms in some provinces of the Mekong Delta, Vietnam. Material and Methods: A total of 203 E. coli isolates from goat feces were examined by PCR for serotypes (O8, O9, O25, O26, O45, O103, O146, O157, and O159), eight antibiotic-resistance genes, four heavy-metal-resistance genes, and four pathogenic genes. Results: By PCR, 20.20% of E. coli isolates belonging to serotypes O8 (6.40%), O45 (13.30%), and O159 (0.49%) were identified. Antibiotic-resistance genes were recorded at high rates in E. coli isolates, especially genes blaampC (98.52%), tetA (50.74%), sulII (34.48%), qnrA (20.69%), and aadA1 (20.69%). Moreover, 55.67% of these E. coli isolates harbored multiple antibiotic-resistance genes. Among heavy-metal-resistance genes, the gene czcD encoding for resistance to cobalt, zinc, and cadmium was the most prevalent (59.11%). In addition, the most frequent virulent gene was stx1 (15.27%), followed by gene stx2 (6.90%), eae, and hlyA (1.48%). Conclusion: These results revealed that goats were a natural reservoir of pathogenic E. coli serotypes, which could cause severe diseases in animals and humans. Moreover, these E. coli isolates showed a high ability to resist diverse antibiotics. Thus, managing the prevalence of pathogenic E. coli is essential for protecting public health in the Mekong Delta. [J Adv Vet Anim Res 2025; 12(2.000): 420-426]

Objectives: The current study sought to ascertain the mRNA expression and establish the complementary DNA (cDNA) sequences of pigeon brain’s glutamate receptor 2B of N-methyl-D-aspartate (GluN2B) type. Material and Methods: Adult pigeons (Columba livia; n = 8, sharing an equal number of males and females) were used. After proper anesthesia, the brain was exposed, and small pieces of cerebellum, optic tectum, thalamus, and telencephalon were collected quickly; total ribonucleic acid (RNA) was isolated, and cDNA was synthesized for PCR amplification. The ABI Prism 3100 Genetic Analyzer was used to analyze the sequences of the corresponding cDNA fragments. Results: In RT-PCR, the findings unequivocally demonstrated that the pigeon brain’s cerebellum, optic tectum, thalamus, and telencephalon all expressed the mRNA for GluN2B. The cDNA sequence of pigeon GluN2B was obtained from PCR-amplified products and included 51 base pairs(bp) of the 5’ untranslated region (UTR), a 4,512-bp open reading frame, and 13 bps of the 3’ UTR. Pigeon GluN2B’s cDNA sequencing displayed 85% identity for human GluN2B and 95% identity for chicken. The amino acid sequences encoded by the pigeon GluN2B gene shared between 85% and 97% similarity with those of humans, rats, and mice. Molecular phylogenetic analysis using the neighbor-joining method showed that pigeon GluN2B is closely related to the GluN2B proteins of these other species. Conclusion: The findings suggest that certain neurons in the pigeon brain GluN2B mRNA. They also indicate the presence of various glutamatergic networks and connections within the avian brain. J. Adv. Vet. Anim. Res., 12(2): 427–432, June 2025 http://doi.org/10.5455/javar.2025.l909

Objectives: The current study sought to ascertain the mRNA expression and establish the complementary DNA (cDNA) sequences of pigeon brain's glutamate receptor 2B of N-methyl-D-aspartate (GluN2B) type. Material and Methods: Adult pigeons (Columba livia; n = 8, sharing an equal number of males and females) were used. After proper anesthesia, the brain was exposed, and small pieces of cerebellum, optic tectum, thalamus, and telencephalon were collected quickly; total ribonucleic acid (RNA) was isolated, and cDNA was synthesized for PCR amplification. The ABI Prism 3100 Genetic Analyzer was used to analyze the sequences of the corresponding cDNA fragments. Results: In RT-PCR, the findings unequivocally demonstrated that the pigeon brain's cerebellum, optic tectum, thalamus, and telencephalon all expressed the mRNA for GluN2B. The cDNA sequence of pigeon GluN2B was obtained from PCR-amplified products and included 51 base pairs (bp) of the 5' untranslated region (UTR), a 4,512-bp open reading frame, and 13 bps of the 3' UTR. Pigeon GluN2B's cDNA sequencing displayed 85% identity for human GluN2B and 95% identity for chicken. The amino acid sequences encoded by the pigeon GluN2B gene shared between 85% and 97% similarity with those of humans, rats, and mice. Molecular phylogenetic analysis using the neighbor-joining method showed that pigeon GluN2B is closely related to the GluN2B proteins of these other species. Conclusion: The findings suggest that certain neurons in the pigeon brain produce GluN2B mRNA. They also indicate the presence of various glutamatergic networks and connections within the avian brain. [J Adv Vet Anim Res 2025; 12(2.000): 427-432]