Objective: Cannabis sativa L. is aware of a rich source of bioactive substances with various structures that exhibit pharmacological activity in the central nervous system, cardiovascular, cerebrovascular, respiratory, reproductive, and gastrointestinal systems. Materials and Methods: In this study, cannabis sugar leaves were soaked in 99% ethanol, followed by evaporation. The antibacterial effect of the cannabis sugar leaf extract was then evaluated using the disc diffusion method. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined using broth dilution. Results: The results of this study indicated that the cannabis sugar leaf extract inhibited Bacillus cereus, Vibrio cholerae, Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis when compared to tetracycline, but it did not inhibit Pseudomonas aeruginosa. The MIC and MBC of the cannabis sugar leaves extract against B. cereus, V. cholerae, E. coli, S. aureus, and S. epidermidis were 0.977, 1.953, 31.25, 62.5, 125, 250, 250, 500, 250, and 500 mg/ml, respectively. The bioactive compounds in cannabis sugar leaf extract were identified using high-performance liquid chromatography. Conclusion: The results indicated that the major bioactive compounds were Δ-9- tetrahydrocannabinol (THC) and cannabidiol (CBD). While minor bioactive compounds included gallic acid and tannic acid. These results support the benefits of cannabis sugar leaf extract, which has been used for its pharmacological properties and may be useful as an alternative antimicrobial agent in medicine. J. Adv. Vet. Anim. Res., 12(1): 44–52, March 2025 http://doi.org/10.5455/javar.2025.l870

Objective: Cannabis sativa L. is aware of a rich source of bioactive substances with various struc¬tures that exhibit pharmacological activity in the central nervous system, cardiovascular, cerebro¬vascular, respiratory, reproductive, and gastrointestinal systems. Materials and Methods: In this study, cannabis sugar leaves were soaked in 99% ethanol, followed by evaporation. The antibacterial effect of the cannabis sugar leaf extract was then evaluated using the disc diffusion method. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined using broth dilution. Results: The results of this study indicated that the cannabis sugar leaf extract inhibited Bacillus cereus, Vibrio cholerae, Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis when compared to tetracycline, but it did not inhibit Pseudomonas aeruginosa. The MIC and MBC of the cannabis sugar leaves extract against B. cereus, V. cholerae, E. coli, S. aureus, and S. epidermidis were 0.977, 1.953, 31.25, 62.5, 125, 250, 250, 500, 250, and 500 mg/ml, respectively. The bioactive compounds in cannabis sugar leaf extract were identified using high-performance liquid chromatography. Conclusion: The results indicated that the major bioactive compounds were Δ-9- tetrahydrocan-nabinol (THC) and cannabidiol (CBD). While minor bioactive compounds included gallic acid and tannic acid. These results support the benefits of cannabis sugar leaf extract, which has been used for its pharmacological properties and may be useful as an alternative antimicrobial agent in medicine. [J Adv Vet Anim Res 2025; 12(1.000): 44-52]

Objective: The increasing antibiotic resistance in poultry pathogens presents a significant public health risk, leading to the exploration of effective alternatives in broiler feed, particularly probiotics such as Bacillus subtilis. This study aimed to isolate B. subtilis strains from indigenous chicken feces that can inhibit Escherichia coli and Salmonella typhimurium strains, common causes of diarrhea in poultry. Materials and Methods: Bacillus strains were isolated from chicken feces and screened for antibacterial activity using an agar well diffusion assay. Bacillus strains were identified via 16S rRNA gene sequencing. Their probiotic potential was assessed through in vitro assays measuring extracellular enzyme production, adhesion properties, and resilience to acidic and bile salt conditions. Freeze-drying techniques were applied to evaluate strain viability and stability. In vivo studies determined the colonization ability of selected strains in the chicken intestine. Results: From 121 B. subtilis isolates, six B. subtilis strains demonstrated notable antibacterial activity against both E. coli and S. typhimurium. Five strains were confirmed as B. subtilis through sequencing. Based on their probiotic attributes, B. subtilis H1 and B. subtilis BSn5 were identified as the most promising candidates. Notably, B. subtilis BSn5 exhibited stable viability when freeze dried, surviving for up to two months, and successfully colonized the chicken intestinal tract in vivo. Conclusion: These findings indicate that B. subtilis BSn5 may serve as a viable probiotic alternative to antibiotics in poultry, with regular supplementation necessary to sustain its benefits. J. Adv. Vet. Anim. Res., 12(1): 53–63, March 2025 http://doi.org/10.5455/javar.2025.l871

Objective: The increasing antibiotic resistance in poultry pathogens presents a significant public health risk, leading to the exploration of effective alternatives in broiler feed, particularly probiotics such as Bacillus subtilis. This study aimed to isolate B. subtilis strains from indigenous chicken feces that can inhibit Escherichia coli and Salmonella typhimurium strains, common causes of diarrhea in poultry. Materials and Methods: Bacillus strains were isolated from chicken feces and screened for antibacterial activity using an agar well diffusion assay. Bacillus strains were identified via 16S rRNA gene sequencing. Their probiotic potential was assessed through in vitro assays measuring extracellular enzyme production, adhesion properties, and resilience to acidic and bile salt conditions. Freeze-drying techniques were applied to evaluate strain viability and stability. In vivo studies determined the colonization ability of selected strains in the chicken intestine. Results: From 121 B. subtilis isolates, six B. subtilis strains demonstrated notable antibacterial activity against both E. coli and S. typhimurium. Five strains were confirmed as B. subtilis through sequencing. Based on their probiotic attributes, B. subtilis H1 and B. subtilis BSn5 were identified as the most promising candidates. Notably, B. subtilis BSn5 exhibited stable viability when freeze-dried, surviving for up to two months, and successfully colonized the chicken intestinal tract in vivo. Conclusion: These findings indicate that B. subtilis BSn5 may serve as a viable probiotic alternative to antibiotics in poultry, with regular supplementation necessary to sustain its benefits. [J Adv Vet Anim Res 2025; 12(1.000): 53-63]

Objective: A cross-sectional study was conducted to reveal the hematology, serum biochemistry, and reproductive behavior of free-roaming cats in the Sylhet Division of Bangladesh. Materials and Methods: Overall, 130 free-roaming cats were captured, of which 52 were Tom (male), 69 were Queen (female), and 9 were kittens. Regarding their ages, the cats were divided into three groups: <1 year as kittens, 1 to 2 years as young, and adults over two years. Thirty blood samples were drawn from the cephalic or saphenous veins of the hind leg for hematological and serum biochemical analysis. Results: The findings revealed that the average hemoglobin concentration was significantly higher in kittens than in young and adult cats (p <1 year as kittens, 1 to 2 years as young, and adults over two years. Thirty blood samples were drawn from the cephalic or saphenous veins of the hind leg for hematological and serum biochemical analysis. Results: The findings revealed that the average hemoglobin concentration was significantly higher in kittens than in young and adult cats (p >< 0.05). In comparison to young and adult cats, kittens had considerably greater red blood cell, packed cell volume, mean corpuscular volume, and platelet counts, whereas mature cats had much higher white blood cell counts. The number of lymphocytes and monocytes was shown to be non-significant, while other components, such as neutrophils, eosinophils, and basophils, varied considerably by age group. Rural cats showed significantly (p < 0.05) higher alanine transaminase and aspartate aminotransferase, whereas urban cats showed higher serum glucose (p < 0.001). The duration of the cat’s estrous cycle was (5.63 ± 1.75) days, as were the inter-estrous phase’s length (7 days), gestation time (66.6 days), and queening rate (65.2). There were 3.92 ± 0.55 kittens per litter. Males reached puberty at a young age of 9.3 months, while females did so at 8.3 months. Conclusion: These findings may contribute to the growing body of research on free-roaming cat populations, which is important for understanding the ecology and behavior of these cats and constructing strategies for their conservation and welfare. J. Adv. Vet. Anim. Res., 12(1): 70–79, March 2025 http://doi.org/10.5455/javar.2025.l873

Objective: A cross-sectional study was conducted to reveal the hematology, serum biochemistry, and reproductive behavior of free-roaming cats in the Sylhet Division of Bangladesh. Materials and Methods: Overall, 130 free-roaming cats were captured, of which 52 were Tom (male), 69 were Queen (female), and 9 were kittens. Regarding their ages, the cats were divided into three groups: [J Adv Vet Anim Res 2025; 12(1.000): 70-79]

Objective: This study aimed to use the potential chitosan processing waste from shrimp heads as a premix to improve the production performance of laying chickens. Materials and Methods: This research uses shrimp head waste, NaOH, hydrochloric acid, Na2SeO3, and NaCl solutions. Processing shrimp head waste into chitosan is done in three ways, namely demineralization, deproteination, and deacetylation. Each of the resulting liquid wastes is then mixed until a neutral mixture is obtained. The data observed were the nutrient and amino acid content of liquid waste from the deproteinization, demineralization, and deacetylation processes, as well as the neutral mixture. Results: The results showed liquid waste from the process of making chitosan from shrimp head waste using deproteination, demineralization, and deacetylation methods, as well as a neutral mixture containing little energy, protein, and fat but is rich in minerals. The most abundant and complete amino acid content is found in deproteination process wastewater and neutral mixtures. Conclusion: In conclusion, the waste from making chitosan from shrimp head waste through deproteinization, demineralization, and deacetylation, and the neutral mixture contains enough minerals and amino acids needed by livestock, so it has the potential to be used as a premix. J. Adv. Vet. Anim. Res., 12(1): 117–122, March 2025 http://doi.org/10.5455/javar.2025.l878

Objective: This study aimed to use the potential chitosan processing waste from shrimp heads as a premix to improve the production performance of laying chickens. Materials and Methods: This research uses shrimp head waste, NaOH, hydrochloric acid, Na₂SeO₃, and NaCl solutions. Processing shrimp head waste into chitosan is done in three ways, namely demineralization, deproteination, and deacetylation. Each of the resulting liquid wastes is then mixed until a neutral mixture is obtained. The data observed were the nutrient and amino acid content of liquid waste from the deproteinization, demineralization, and deacetylation processes, as well as the neutral mixture. Results: The results showed liquid waste from the process of making chitosan from shrimp head waste using deproteination, demineralization, and deacetylation methods, as well as a neutral mixture containing little energy, protein, and fat but is rich in minerals. The most abundant and complete amino acid content is found in deproteination process wastewater and neutral mixtures. Conclusion: In conclusion, the waste from making chitosan from shrimp head waste through deproteinization, demineralization, and deacetylation, and the neutral mixture contains enough minerals and amino acids needed by livestock, so it has the potential to be used as a premix. [J Adv Vet Anim Res 2025; 12(1.000): 117-122]

Objective: This study aimed to assess Aerodramus fuciphagus (Thunberg, 1812) nest’s mineral content, cytotoxicity effects, and antioxidant activity. The Edible birds’ nests (EBNs) were obtained from Tanah Merah (TM), Tumpat (TU), and Jeli (JE) districts in Kelantan, Malaysia. Materials and Methods: The mineral content was assessed via inductively coupled plasma mass spectrometry. The cytotoxicity effect was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide and brine shrimp lethality assays. The antioxidant activity was investigated by 2,2´-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and ferric-reducing antioxidant power (FRAP) assays. Results: Valuable macro- and microelements, including sodium, potassium, aluminum, calcium, magnesium, zinc, iron, and selenium, were detected in the EBN samples. The cytotoxicity test revealed that JE hydrochloric acid (HCl) extract had promising efficacy against HeLa cell lines, with IC50 values of 20.00 ± 0.57 µg/ml. Similarly, TU sodium hydroxide (NaOH) extract inhibited the proliferation of MCF-7 cell lines (IC50 0.03 ± 0.01 µg/ml). Based on the BSL assay, the EBNs were considered safe for consumption as the LC50 values of all the samples were below the toxic level (>1,000 µg/ml). TM EBNs aqueous, TU aqueous, and HCl extracts showed promising antioxidant activity at IC50 values (10 µg/ml) in the DPPH assay. However, the NaOH extract from TU exhibited the highest FRAP value (1.38 ± 0.00 mM gallic acid equivalents gm⁻¹) compared to all other extracts. Conclusion: This study’s findings demonstrated the potential of EBN as an alternative anticancer agent and natural antioxidant. J. Adv. Vet. Anim. Res., 12(1): 132–140, March 2025 http://doi.org/10.5455/javar.2025.l880

Objective: This study aimed to assess Aerodramus fuciphagus (Thunberg, 1812) nest's mineral content, cytotoxicity effects, and antioxidant activity. The Edible birds' nests (EBNs) were obtained from Tanah Merah (TM), Tumpat (TU), and Jeli (JE) districts in Kelantan, Malaysia. Materials and Methods: The mineral content was assessed via inductively coupled plasma mass spectrometry. The cytotoxicity effect was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide and brine shrimp lethality assays. The antioxidant activity was investigated by 2,2´-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and ferric-reducing antioxidant power (FRAP) assays. Results: Valuable macro- and microelements, including sodium, potassium, aluminum, calcium, magnesium, zinc, iron, and selenium, were detected in the EBN samples. The cytotoxicity test revealed that JE hydrochloric acid (HCl) extract had promising efficacy against HeLa cell lines, with IC50 values of 20.00 ± 0.57 μg/ml. Similarly, TU sodium hydroxide (NaOH) extract inhibited the proliferation of MCF-7 cell lines (IC50 0.03 ± 0.01 μg/ml). Based on the BSL assay, the EBNs were considered safe for consumption as the LC50 values of all the samples were below the toxic level (>1,000 μg/ml). TM EBNs aqueous, TU aqueous, and HCl extracts showed promising antioxidant activity at IC50 values (10 μg/ml) in the DPPH assay. However, the NaOH extract from TU exhib¬ited the highest FRAP value (1.38 ± 0.00 mM gallic acid equivalents gm⁻¹) compared to all other extracts. Conclusion: This study's findings demonstrated the potential of EBN as an alternative anticancer agent and natural antioxidant. [J Adv Vet Anim Res 2025; 12(1.000): 132-140]

Objectives: The primary objective of this study is to provide a comprehensive analysis of the differences between conventional and brown midrib (BMR) sorghum in terms of biomass, nutrient quality, nutrient digestibility, and animal performance. Materials and Methods: A comprehensive database was created by integrating 73 datasets from 29 articles. The different studies were denoted as random effects, while the BMR sorghum variety was described as a fixed component. Afterward, these two aspects were calculated utilizing a linear mixed model. Results: According to the findings of the present meta-analysis, conventional sorghum produces higher quantities of forage and grain biomass in comparison to BMR (p < 0.001). As expected, BMR sorghum has a lower content of neutral detergent fiber (NDF), acid detergent fiber, and lignin compared to conventional sorghum (p < 0.001). The digestibility of BMR sorghum performed better than conventional sorghum (p < 0.001), especially for NDF digestibility (54.98% vs. 47.37%). BMR shows suitability as a fodder option for dairy cows due to its superior milk yield compared to conventional sorghum (15.04 vs. 14.06 kg/day; p < 0.01). Conclusion: In conclusion, BMR sorghum produces higher biomass compared to conventional sorghum. Nevertheless, in terms of nutrient quality and digestibility, BMR sorghum is the most optimal choice. The results will significantly improve animal performance. J. Adv. Vet. Anim. Res., 12(1): 157–168, March 2025 http://doi.org/10.5455/javar.2025.l883

Objectives: The primary objective of this study is to provide a comprehensive analysis of the differences between conventional and brown midrib (BMR) Sorghum in terms of biomass, nutrient quality, nutrient digestibility, and animal performance. Materials and Methods: A comprehensive database was created by integrating 73 datasets from 29 articles. The different studies were denoted as random effects, while the BMR Sorghum variety was described as a fixed component. Afterward, these two aspects were calculated utilizing a linear mixed model. Results: According to the findings of the present meta-analysis, conventional Sorghum produces higher quantities of forage and grain biomass in comparison to BMR (p < 0.001). As expected, BMR Sorghum has a lower content of neutral detergent fiber (NDF), acid detergent fiber, and lignin compared to conventional Sorghum (p < 0.001). The digestibility of BMR Sorghum performed better than conventional Sorghum (p < 0.001), especially for NDF digestibility (54.98% vs. 47.37%). BMR shows suitability as a fodder option for dairy cows due to its superior milk yield compared to conventional Sorghum (15.04 vs. 14.06 kg/day; p < 0.01). Conclusion: In conclusion, BMR Sorghum produces higher biomass compared to conventional Sorghum. Nevertheless, in terms of nutrient quality and digestibility, BMR Sorghum is the most optimal choice. The results will significantly improve animal performance. [J Adv Vet Anim Res 2025; 12(1.000): 157-168]

Objective: This study aims to assess the impact of aging time on the quality of meat, fatty acid profiles, and volatile compounds in cooked Thai native beef. Materials and Methods: The experiment utilized a randomized complete block design, where the aging time (0, 7, 14, 21, and 28 days) served as the treatment and the longissimus thoracis et lumborum muscle from Thai native cattle (n = 4) was considered a block. Results: Meat color and Warner-Bratzler shear force values decreased as the aging time increased. Furthermore, the moisture content decreased while the protein and ash content increased with a longer aging time. The proportions of fatty acids showed significant differences with varying aging times. These fatty acids exhibited the highest proportions in the 14-day dry-aged beef samples. The levels of short-chain aldehydes (pentanal, heptanal, octanal, nonanal, decanal, and 2-nonenal) and alcohols (1-hexanol and 1-octanol) decreased as the aging time increased. However, there was an increase in long-chain aldehydes (tetradecanal, hexadecanal, heptadecanal, and octadecanal) with longer aging time. In addition, the levels of toluene, dodecane, tridecane, methyl-pyrazine, 2,5-dimethyl-pyrazine, trimethyl-pyrazine, and dimethyl trisulfide were higher in 14-day dry-aged beef compared to beef that had not been aged. Furthermore, a correlation was found between the number of the 6 significant fatty acids in the dry-aged beef samples and the 12 volatile compounds in cooked beef. Conclusion: Our results suggest that aging times significantly influence the fatty acid profiles of Thai native beef, which in turn are correlated with the characteristic volatile compounds. J. Adv. Vet. Anim. Res., 12(1): 179–191, March 2025 http://doi.org/10.5455/javar.2025.l885

Objective: This study aims to assess the impact of aging time on the quality of meat, fatty acid profiles, and volatile compounds in cooked Thai native beef. Materials and Methods: The experiment utilized a randomized complete block design, where the aging time (0, 7, 14, 21, and 28 days) served as the treatment and the longissimus thoracis et lumborum muscle from Thai native cattle (n = 4) was considered a block. Results: Meat color and Warner-Bratzler shear force values decreased as the aging time increased. Furthermore, the moisture content decreased while the protein and ash content increased with a longer aging time. The proportions of fatty acids showed significant differences with varying aging times. These fatty acids exhibited the highest proportions in the 14-day dry-aged beef samples. The levels of short-chain aldehydes (pentanal, heptanal, octanal, nonanal, decanal, and 2-non¬enal) and alcohols (1-hexanol and 1-octanol) decreased as the aging time increased. However, there was an increase in long-chain aldehydes (tetradecanal, hexadecanal, heptadecanal, and octadecanal) with longer aging time. In addition, the levels of toluene, dodecane, tridecane, methyl-pyrazine, 2,5-dimethyl-pyrazine, trimethyl-pyrazine, and dimethyl trisulfide were higher in 14-day dry-aged beef compared to beef that had not been aged. Furthermore, a correlation was found between the number of the 6 significant fatty acids in the dry-aged beef samples and the 12 volatile compounds in cooked beef. Conclusion: Our results suggest that aging times significantly influence the fatty acid profiles of Thai native beef, which in turn are correlated with the characteristic volatile compounds. [J Adv Vet Anim Res 2025; 12(1.000): 179-191]

Objective: The objectives of this study are to measure the body weight (BW) and morphometric parameters of Napu buffaloes, identify the growth hormone (GH) and insulin-like growth factor-1 (IGF-1) gene polymorphisms, and associate them with the BW and morphometric parameters of Napu buffaloes. Materials and Methods: This study used 39 Napu buffaloes (9 males and 30 females). Morphometric measurements were performed using a measuring tape. The GH and IGF-1 gene diversity analysis was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing techniques. Results: The results showed that there is diversity in BW and body size of Napu buffaloes due to the influence of age and sex. The results of PCR-RFLP analysis of the GH|AluI gene showed 0% VV, 100% LL, and 0% VL genotypes. While the IGF-1|SnaBI gene shows the genotype AA 0%, BB 100%, and AB 0%. The sequencing results of the GH and IGF-1 genes in Napu buffaloes did not find mutations. Conclusion: Age and sex in Napu buffaloes may affect BW and body size. The GH and IGF-1 genes in Napu buffaloes are monomorphic, so no association can be made with the morphometric parameters of Napu buffaloes. J. Adv. Vet. Anim. Res., 12(1): 205–213, March 2025 http://doi.org/10.5455/javar.2025.l887

Objective: The objectives of this study are to measure the body weight (BW) and morphometric parameters of Napu buffaloes, identify the growth hormone (GH) and insulin-like growth factor-1 (IGF-1) gene polymorphisms, and associate them with the BW and morphometric parameters of Napu buffaloes. Materials and Methods: This study used 39 Napu buffaloes (9 males and 30 females). Morphometric measurements were performed using a measuring tape. The GH and IGF-1 gene diversity analysis was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing techniques. Results: The results showed that there is diversity in BW and body size of Napu buffaloes due to the influence of age and sex. The results of PCR-RFLP analysis of the GH|AluI gene showed 0% VV, 100% LL, and 0% VL genotypes. While the IGF-1|SnaBI gene shows the genotype AA 0%, BB 100%, and AB 0%. The sequencing results of the GH and IGF-1 genes in Napu buffaloes did not find mutations. Conclusion: Age and sex in Napu buffaloes may affect BW and body size. The GH and IGF-1 genes in Napu buffaloes are monomorphic, so no association can be made with the morphometric parameters of Napu buffaloes. [J Adv Vet Anim Res 2025; 12(1.000): 205-213]

Objective: The present study was designed to conduct molecular and morphological identification of cat fleas (Ctenocephalides felis) from Bangladesh along with nucleotide polymorphism and phylogenetic analysis. Materials and Methods: Samples were collected from two hosts (cat and human). The species was identified through morphological studies first, and then DNA was extracted for subsequent molecular analysis. A part of the mitochondrial 16S rRNA gene was amplified by polymerase chain reaction using extracted DNA as a template. The amplified region was sequenced using the Sanger dideoxy method. The sequence was subjected to NCBI BLASTn search. BioEdit and MEGA 11 software were used for multiple sequence alignment (MSA) and generating a phylogenetic tree. Results: Morphological features such as shape, size, and appendages showed similarity with C. felis. The metatibial formula of chaetotaxy (2-2-2-2-1-3) was confirmed for species-level identification. NCBI BLASTn search showed the highest sequence identity with the available sequence of C. felis such as 99.78% (NC_049858.1) and 99.12% (MW420044.1 and MK941844.1) with 100% query coverage. MSA of C. felis sequences from different geographical distributions show their sequence affinities with each other, and the phylogenetic tree presents their relationship with each other. Conclusion: Both morphological and molecular studies clearly indicate the identity and confirmation of cat flea (C. felis) from Bangladesh. J. Adv. Vet. Anim. Res., 12(1): 280–286, March 2025 http://doi.org/10.5455/javar.2025.l894

Objective: The present study was designed to conduct molecular and morphological identification of cat fleas (Ctenocephalides felis) from Bangladesh along with nucleotide polymorphism and phylogenetic analysis. Materials and Methods: Samples were collected from two hosts (cat and human). The species was identified through morphological studies first, and then DNA was extracted for subsequent molecular analysis. A part of the mitochondrial 16S rRNA gene was amplified by polymerase chain reaction using extracted DNA as a template. The amplified region was sequenced using the Sanger dideoxy method. The sequence was subjected to NCBI BLASTn search. BioEdit and MEGA 11 software were used for multiple sequence alignment (MSA) and generating a phylogenetic tree. Results: Morphological features such as shape, size, and appendages showed similarity with C. felis. The metatibial formula of chaetotaxy (2-2-2-2-1-3) was confirmed for species-level identification. NCBI BLASTn search showed the highest sequence identity with the available sequence of C. felis such as 99.78% (NC_049858.1) and 99.12% (MW420044.1 and MK941844.1) with 100% query coverage. MSA of C. felis sequences from different geographical distributions show their sequence affinities with each other, and the phylogenetic tree presents their relationship with each other. Conclusion: Both morphological and molecular studies clearly indicate the identity and confirmation of cat flea (C. felis) from Bangladesh. [J Adv Vet Anim Res 2025; 12(1.000): 280-286]