Objective: The present study was aimed to evaluate dose-dependent effects of phytobiotic (Galibiotic) supplements in feed on growth performance, hematological parameters, intestinal pH, and gut bacterial population in broiler chick. Materials and Methods: A total of 50 ten day old broiler chicks were divided into five groups, namely, Group A as control (without galibiotic), Group B (galibiotic at 1 gm/kg feed), Group C (gal­ibiotic at 2 gm/kg feed), Group D (galibiotic at 5 gm/kg feed), and Group E (galibiotic at 10 gm/kg feed). All the birds were reared for 42 days and samples were collected before and after sacrifice. Results: Live body weights showed no significant differences between the groups but overall feed conversion ratios (FCRs) of treatment groups were significantly low in Group E having the lowest. Blood samples collected for hematology differed significantly (p < 0.01) among the different groups. Intestinal pH was lower in treatment groups with Group E having the lowest. Cecal total viable count was highest in Group A and lowest in Group E. The cecal coliform count was low in all the treatment groups. Conclusion: Along with previously published report, it may conclude that the phytobiotic could be used as an alternative to antibiotics due to positive growth performance, lower FCR, carcass quality, and improved gut health of broiler chicks. J. Adv. Vet. Anim. Res., 6(2): 253-259, June 2019

Objective: The present study was aimed to evaluate dose-dependent effects of phytobiotic (Galibiotic) supplements in feed on growth performance, hematological parameters, intestinal pH, and gut bacterial population in broiler chick. Materials and Methods: A total of 50 ten day old broiler chicks were divided into five groups, namely, Group A as control (without galibiotic), Group B (galibiotic at 1 gm/kg feed), Group C (gal¬ibiotic at 2 gm/kg feed), Group D (galibiotic at 5 gm/kg feed), and Group E (galibiotic at 10 gm/kg feed). All the birds were reared for 42 days and samples were collected before and after sacrifice. Results: Live body weights showed no significant differences between the groups but overall feed conversion ratios (FCRs) of treatment groups were significantly low in Group E having the lowest. Blood samples collected for hematology differed significantly (p < 0.01) among the different groups. Intestinal pH was lower in treatment groups with Group E having the lowest. Cecal total viable count was highest in Group A and lowest in Group E. The cecal coliform count was low in all the treatment groups. Conclusion: Along with previously published report, it may conclude that the phytobiotic could be used as an alternative to antibiotics due to positive growth performance, lower FCR, carcass quality, and improved gut health of broiler chicks. [J Adv Vet Anim Res 2019; 6(2.000): 253-259]

Objective: This research aims to study the effectiveness of landmark- and semi-landmark-based geometric morphometric (LMSL) in the identification of four Culex vectors, namely, C. quinquefas­ciatus, C. visnui, C. sitiens, and C. whitmorei in Thailand and also compared the potential between LMSL and the landmark-based geometric morphometric method (LM). Materials and Methods: The overall size of the wing sample was estimated by the centroid size. Shape variables were computed as principal components of the “partial warp” calculated after generalized procrustes analysis of raw coordinates. Discriminant analysis of the canonical variables performed to explore the shape dissimilarity between Culex species has been shown as a factor map and to calculate the Mahalanobis distance. Size and shape differences based on pairwise Mahalanobis distances were tested using non-parametric methods (1,000 cycles) with Bonferroni correction at a p-value of <0.05. Results: A total of 120 individuals were used that were divided into 30 individuals per Culex species. The mean CS of C. sitiens had the largest wings followed by C. visnui, C. quinquefasciatus, and C. whitmorei in LM and LMSM. The patterns of statistical difference in CS of both methods were similar and wing shapes among Culex species were different based on a comparison of pairwise Mahalanobis distances (p < 0.05) in both methods. For the cross-validated reclassification test, LM provided Culex species separation ranging from 54% to 84% and 51% to 93% for LMSM. Conclusion: Thus, LMSM is another option to use for the identification in mosquito vectors that have a curved line on the wing specific to the species. J. Adv. Vet. Anim. Res., 6(3): 278-283, September 2019

Objective: This research aims to study the effectiveness of landmark- and semi-landmark-based geometric morphometric (LMSL) in the identification of four Culex vectors, namely, C. quinquefas¬ciatus, C. visnui, C. sitiens, and C. whitmorei in Thailand and also compared the potential between LMSL and the landmark-based geometric morphometric method (LM). Materials and Methods: The overall size of the wing sample was estimated by the centroid size. Shape variables were computed as principal components of the partial warp calculated after generalized procrustes analysis of raw coordinates. Discriminant analysis of the canonical variables performed to explore the shape dissimilarity between Culex species has been shown as a factor map and to calculate the Mahalanobis distance. Size and shape differences based on pairwise Mahalanobis distances were tested using non-parametric methods (1,000 cycles) with Bonferroni correction at a p-value of [J Adv Vet Anim Res 2019; 6(3.000): 278-283]

Objective: Major histocompatibility complex (MHC) is a set of molecular proteins on the surface of antigen presenting cells encoded by a large gene family which are important parts of the immune system. This study was conducted to convey information on the genetic characteristics of the MHC II DRB3 gene in riverine and swamp buffaloes. Materials and Methods: Characterization of MHC II DRB3 gene was carried out using polymerase chain reaction (PCR)-based assay. Thirty-milliliter milk samples were collected from 10 swamp-type and 10 riverine-type buffaloes. RNA from milk samples were extracted using Trizol and then followed by reverse transcription-PCR (RT-PCR). Results: The phylogenetic analysis with 1,000 bootstrap replications clearly showed complex parsimony in MHC II DRB3 gene between 10 riverine- and 10 swamp-type but also confirmed that the samples are similar to Bubalus bubalis. Aligned sequences of the 20 water buffaloes were compared with three other ruminants (Bos taurus, Ovis aries, and Capra hircus) and non-ruminant (Sus scrofa) that serve as an outgroup. MHC sequences from GenBank show that there was an average of 705 identical pairs, with 22 transitional pairs and 30 transversional pairs with a ratio of 0.7. Conclusion: Based on the molecular data, the current study conforms to other works of literature that this gene is highly polymorphic which can be due to its function in the immune responsiveness and disease resistance. Further study on the immunological response of MHC II DRB3 to infection may elucidate its underlying function and role in the protection against specific disease of animals. J. Adv. Vet. Anim. Res., 6(3): 308-314, September 2019

Objective: Major histocompatibility complex (MHC) is a set of molecular proteins on the surface of antigen presenting cells encoded by a large gene family which are important parts of the immune system. This study was conducted to convey information on the genetic characteristics of the MHC II DRB3 gene in riverine and swamp buffaloes. Materials and Methods: Characterization of MHC II DRB3 gene was carried out using polymerase chain reaction (PCR)-based assay. Thirty-milliliter milk samples were collected from 10 swamp-type and 10 riverine-type buffaloes. RNA from milk samples were extracted using Trizol and then followed by reverse transcription-PCR (RT-PCR). Results: The phylogenetic analysis with 1,000 bootstrap replications clearly showed complex parsimony in MHC II DRB3 gene between 10 riverine- and 10 swamp-type but also confirmed that the samples are similar to Bubalus bubalis. Aligned sequences of the 20 water buffaloes were compared with three other ruminants (Bos taurus, Ovis aries, and Capra hircus) and non-ruminant (Sus scrofa) that serve as an outgroup. MHC sequences from GenBank show that there was an average of 705 identical pairs, with 22 transitional pairs and 30 transversional pairs with a ratio of 0.7. Conclusion: Based on the molecular data, the current study conforms to other works of literature that this gene is highly polymorphic which can be due to its function in the immune responsiveness and disease resistance. Further study on the immunological response of MHC II DRB3 to infection may elucidate its underlying function and role in the protection against specific disease of animals. [J Adv Vet Anim Res 2019; 6(3.000): 308-314]

Objective: In this study, we investigated the potential immune-enhancing effects in addition to anti-oxidative stress properties of commercially accessible Bacillus subtilis supplementation in Barki lambs. Materials and Methods: Twenty apparently healthy weaned Barki lambs were used in this study and distributed randomly into two experimental groups: Negative control group, received control basal diet without any feed supplements and a supplemented group, received control basal diet supplemented with water added to commercially accessible bacilli at 1 gm/l/day for 30 consecutive days. Blood samples were collected from each lamb before starting the experiment (T0), 2 weeks (T15), and 4 weeks (T30) post-supplementation for serum biochemical analyses, total leucocytes and lymphocytes count, and real-time polymerase chain reaction assays. Results: The supplemented group showed a significant increase (p < 0.05) in the total number of leukocytes and the number of lymphocytes, lysozyme activity, reduced glutathione, total antioxidant capacity with a significantly lower malondialdehyde values at T30 and significantly higher levels (p < 0.05) of serum catalase and nitric oxide at T15 as compared with control ones. B. subtilis elicited maximal up-regulation of most of the studied genes compared with the control group. Conclusion: The results herein suggest that B. subtilis could be used as useful nutritional supplements to support the immune system in healthy lambs. J. Adv. Vet. Anim. Res., 6(3): 333-340, September 2019

Objective: In this study, we investigated the potential immune-enhancing effects in addition to anti-oxidative stress properties of commercially accessible Bacillus subtilis supplementation in Barki lambs. Materials and Methods: Twenty apparently healthy weaned Barki lambs were used in this study and distributed randomly into two experimental groups: Negative control group, received control basal diet without any feed supplements and a supplemented group, received control basal diet supplemented with water added to commercially accessible bacilli at 1 gm/l/day for 30 consecutive days. Blood samples were collected from each lamb before starting the experiment (T0), 2 weeks (T15), and 4 weeks (T30) post-supplementation for serum biochemical analyses, total leucocytes and lymphocytes count, and real-time polymerase chain reaction assays. Results: The supplemented group showed a significant increase (p < 0.05) in the total number of leukocytes and the number of lymphocytes, lysozyme activity, reduced glutathione, total antioxidant capacity with a significantly lower malondialdehyde values at T30 and significantly higher levels (p < 0.05) of serum catalase and nitric oxide at T15 as compared with control ones. B. subtilis elicited maximal up-regulation of most of the studied genes compared with the control group. Conclusion: The results herein suggest that B. subtilis could be used as useful nutritional supplements to support the immune system in healthy lambs. [J Adv Vet Anim Res 2019; 6(3.000): 333-340]

Objective: The objective of the study was to investigate Foot and Mouth Disease virus (FMDV) outbreak in cattle in the Sarankhola Upazila under Bagerhat district of Bangladesh with isolation, identification, and molecular characterization of FMDV during April 2018. Materials and Methods: This Upazila is located at southern border of Bangladesh and surrounded by mangrove forest Sundarban. The outbreak investigation team collected epidemiological data from outbreak location. In addition, the team collected a total of 30 (15 calves, 15 adult) tongue epithelial tissue samples from a clinically FMD-affected cattle. The confirmation of FMDV and its three serotypes (A, O, and Asia-1) was performed by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). An amplified product of the VP1 region of FMDV genome was sequenced by Sanger sequencing method after cultivation and reconfirmation of FMDV into the BHK21 cell line. Genetic variability was studied by constructing a phylogenetic tree. Results: The investigation survey was carried out in overall 8,393 (8,393/15,580; 53.89%) cases including 3,050 (3,050/8,393; 36.34%) cases in calf and 5,343 (5,343/8,393; 59.77%) cases in adult cattle. The overall case fatality rate (CFR) was recorded as 2.27% (354/15,580) with significantly highest CFR in the calf (71.46%; 253/354) compared to an adult. The collected all 30 samples found with FMDV positive and mixed infection of all samples with serotype Asia-1 and serotype O were observed. In BHK 21 cell line, the eight FMDV positive samples showed a typical cytopathic effect during the third passage. Finally, DNA sequence data of two isolates found closely related with the isolates of bordering country India and Myanmar. Conclusion: The investigation identified the risk factors involved in an outbreak of FMDV, namely, sharing the common paddy land after harvesting, no FMD vaccination, the interaction between cattle and wildlife, and cross bordering movement. J. Adv. Vet. Anim. Res., 6(3): 346-354, September 2019

Objective: The objective of the study was to investigate Foot and Mouth Disease virus (FMDV) outbreak in cattle in the Sarankhola Upazila under Bagerhat district of Bangladesh with isolation, identification, and molecular characterization of FMDV during April 2018. Materials and Methods: This Upazila is located at southern border of Bangladesh and surrounded by mangrove forest Sundarban. The outbreak investigation team collected epidemiological data from outbreak location. In addition, the team collected a total of 30 (15 calves, 15 adult) tongue epithelial tissue samples from a clinically FMD-affected cattle. The confirmation of FMDV and its three serotypes (A, O, and Asia-1) was performed by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). An amplified product of the VP1 region of FMDV genome was sequenced by Sanger sequencing method after cultivation and reconfirmation of FMDV into the BHK21 cell line. Genetic variability was studied by constructing a phylogenetic tree. Results: The investigation survey was carried out in overall 8,393 (8,393/15,580; 53.89%) cases including 3,050 (3,050/8,393; 36.34%) cases in calf and 5,343 (5,343/8,393; 59.77%) cases in adult cattle. The overall case fatality rate (CFR) was recorded as 2.27% (354/15,580) with significantly highest CFR in the calf (71.46%; 253/354) compared to an adult. The collected all 30 samples found with FMDV positive and mixed infection of all samples with serotype Asia-1 and serotype O were observed. In BHK 21 cell line, the eight FMDV positive samples showed a typical cytopathic effect during the third passage. Finally, DNA sequence data of two isolates found closely related with the isolates of bordering country India and Myanmar. Conclusion: The investigation identified the risk factors involved in an outbreak of FMDV, namely, sharing the common paddy land after harvesting, no FMD vaccination, the interaction between cattle and wildlife, and cross bordering movement. [J Adv Vet Anim Res 2019; 6(3.000): 346-354]

Objectives: Drug residues in poultry products could lead to the development of antibiotic-resistant bacteria as in any living animal and human alike. Extensive use of antibiotics in animals to promote growth rate, increase feed efficiency, and for prevention of intestinal infections has led to the development of resistant bacteria in the gastrointestinal tract. The present study was conducted to evaluate the effects of biological supplementation of probiotic, phytobiotic, and their combination over antibiotic on growth performance, microbial load, and hematological parameters in Broiler. Materials and methods: Sixty-five broiler chicken were divided into five groups (12 birds in each group), namely, group A (basal diet), group B (antibiotic, Renamycin 100®), group C (phytobiotic, Galibiotic®), group D (probiotic, Bio-Top®), and group E (combination, Galibiotic®+Galibiotic®) and five were sacrificed for baseline data on day 0. Results: Average final live weight gain was highest in group D (probiotic) than other groups. The feed conversion ratio was highest in group A and lowest in the probiotic group (group C). Blood samples were collected on 14th and 28th day for hematological studies. The mean hematology values regarding the total erythrocyte count, hemoglobin concentration, packed cell volume, and erythrocyte sedimentation rate differed significantly (p < 0.05) among groups. The pH of all the treatment groups was significantly decreased compared to the control group (p < 0.05) where group C was significantly (p < 0.05) lower than all other groups. Highest total viable cell count was observed in control (group A) and total coliform count in phytobiotic (group C) was significantly lower than in other treatment groups (p < 0.05). Conclusion: It may conclude that biological supplements have a significant positive impact on growth performance, hematological parameters, and gut microbial load in broiler chicken of which the probiotic showing the best effects. Supplementation of probiotic in feed could be one of the best candidates as an alternative to antibiotics as growth promoter for safe broiler production. J. Adv. Vet. Anim. Res., 6(3): 409-415, September 2019

Objectives: Drug residues in poultry products could lead to the development of antibiotic-resistant bacteria as in any living animal and human alike. Extensive use of antibiotics in animals to promote growth rate, increase feed efficiency, and for prevention of intestinal infections has led to the development of resistant bacteria in the gastrointestinal tract. The present study was conducted to evaluate the effects of biological supplementation of probiotic, phytobiotic, and their combination over antibiotic on growth performance, microbial load, and hematological parameters in Broiler. Materials and methods: Sixty-five broiler chicken were divided into five groups (12 birds in each group), namely, group A (basal diet), group B (antibiotic, Renamycin 100®), group C (phytobiotic, Galibiotic®), group D (probiotic, Bio-Top®), and group E (combination, Galibiotic®+Galibiotic®) and five were sacrificed for baseline data on day 0. Results: Average final live weight gain was highest in group D (probiotic) than other groups. The feed conversion ratio was highest in group A and lowest in the probiotic group (group C). Blood samples were collected on 14th and 28th day for hematological studies. The mean hematology values regarding the total erythrocyte count, hemoglobin concentration, packed cell volume, and erythrocyte sedimentation rate differed significantly (p < 0.05) among groups. The pH of all the treatment groups was significantly decreased compared to the control group (p < 0.05) where group C was significantly (p < 0.05) lower than all other groups. Highest total viable cell count was observed in control (group A) and total coliform count in phytobiotic (group C) was significantly lower than in other treatment groups (p < 0.05). Conclusion: It may conclude that biological supplements have a significant positive impact on growth performance, hematological parameters, and gut microbial load in broiler chicken of which the probiotic showing the best effects. Supplementation of probiotic in feed could be one of the best candidates as an alternative to antibiotics as growth promoter for safe broiler production. [J Adv Vet Anim Res 2019; 6(3.000): 409-415]

Objective: The study was carried out to develop a wet litter model with magnesium chloride to assess the effectiveness of a polyherbal formulation (PHF) on growth performance, litter and cecal moisture (LCM) level, cecal consistency (CC) score, and footpad lesions (FPLs) score in Ross 308 broiler chickens. Materials and Methods: 1,200 one-day-old chicks were assigned into five groups: normal control, negative control [NTC; treated with 1.7% magnesium chloride hexahydrate (MgCl2.6H2O)], and three treatment groups, T1, T2, and T3, where 750, 1,000, and 2,000 gm/ton of PHF, respectively, were supplemented. All the groups were fed a basal diet until day 7. However, the NTC and treat­ment groups were fed a diet with MgCl2 from days 8 to 42. Results: The addition of MgCl2 for 35 days worsened the growth performance traits in broilers and induced wet litter problems (FPL, high LCM, and poor CC) in the NTC group. However, PHF (750, 1,000, and 2,000 gm/ton) ameliorated the negative effect of a diet with MgCl2 on growth perfor­mance and wet litter problems, but a better response with respect to LCM and CC was observed in 2,000 gm/ton of PHF group, followed by that in 1,000 gm/ton of PHF group and 750 gm/ton of PHF group on day 42. Conclusion: The wet litter broiler model was developed through excessive feeding of MgCl2, which caused the performance parameters to worsen and the emergence of problems associated with the wet litter. Supplementation with PHF ameliorated these problems and, therefore, it can be used for the management of wet litter in poultry. J. Adv. Vet. Anim. Res., 6(4): 536-543, December 2019

Objective: The study was carried out to develop a wet litter model with magnesium chloride to assess the effectiveness of a polyherbal formulation (PHF) on growth performance, litter and cecal moisture (LCM) level, cecal consistency (CC) score, and footpad lesions (FPLs) score in Ross 308 broiler chickens. Materials and Methods: 1,200 one-day-old chicks were assigned into five groups: normal control, negative control [NTC; treated with 1.7% magnesium chloride hexahydrate (MgCl2.6H2O)], and three treatment groups, T1, T2, and T3, where 750, 1,000, and 2,000 gm/ton of PHF, respectively, were supplemented. All the groups were fed a basal diet until day 7. However, the NTC and treat¬ment groups were fed a diet with MgCl2 from days 8 to 42. Results: The addition of MgCl2 for 35 days worsened the growth performance traits in broilers and induced wet litter problems (FPL, high LCM, and poor CC) in the NTC group. However, PHF (750, 1,000, and 2,000 gm/ton) ameliorated the negative effect of a diet with MgCl2 on growth perfor¬mance and wet litter problems, but a better response with respect to LCM and CC was observed in 2,000 gm/ton of PHF group, followed by that in 1,000 gm/ton of PHF group and 750 gm/ton of PHF group on day 42. Conclusion: The wet litter broiler model was developed through excessive feeding of MgCl2, which caused the performance parameters to worsen and the emergence of problems associated with the wet litter. Supplementation with PHF ameliorated these problems and, therefore, it can be used for the management of wet litter in poultry. [J Adv Vet Anim Res 2019; 6(4.000): 536-543]

Objective: This study was conducted to investigate different respiratory diseases in broiler and sonali birds in some selected districts of Bangladesh. Materials and Methods: We were collected a total of 460 blood samples from 46 farms with 36 broiler farms and 10 sonali farms (cross-breed) from 2015 to 2017. All the collected serum sam­ples were tested for determining specific antibodies of avian rhinotracheitis (ART) virus, infectious laryngotracheitis (ILT) virus, infectious bronchitis (IBV) virus, and Ornithobacterium rhinotracheale (ORT) infection using commercially available enzyme-linked immunosorbent assay kits. Results: The overall seropositivity was highest in ORT (45.9%), followed by IBV (37.6%), ART (2.6%), and ILT (0.4%). Out of 360 broiler samples, highest seropositivity was recorded in ORT (43.3%) and lowest in IBV (31.4%). Surprisingly, no broiler samples were found positive for ART and ILT. In case of sonali, the seropositivity was highest in IBV (60%) and lowest in ILT (2%). With respect to types of birds and age groups, the seropositive percentage of all four pathogens was found higher in sonali than broiler. Between two age groups of sonali, the seropositive percentage of ART (12%), ORT (55%), ILT (2%), and IBV (60%) was highest at 21–60 weeks of age compared to 5–20 weeks of age. However, based on location, the seropositive of ORT and IBV was highest in Jamalpur (63.3%) and Fulbariya and Trishal (50%) and lowest in Sreepur (16.7%) and Jamalpur (3.3%). Conclusion: The four pathogens are ubiquitous in nature for the sonali chickens, and the prev­alence of ORT and IBV was the most prevalent viruses in the study areas. This study indicates a need for improved surveillance and characterization of ORT and ART circulating in all types of poultry in Bangladesh. J. Adv. Vet. Anim. Res., 6(4): 561-566, December 2019

Objective: This study was conducted to investigate different respiratory diseases in broiler and sonali birds in some selected districts of Bangladesh. Materials and Methods: We were collected a total of 460 blood samples from 46 farms with 36 broiler farms and 10 sonali farms (cross-breed) from 2015 to 2017. All the collected serum sam¬ples were tested for determining specific antibodies of avian rhinotracheitis (ART) virus, infectious laryngotracheitis (ILT) virus, infectious bronchitis (IBV) virus, and Ornithobacterium rhinotracheale (ORT) infection using commercially available enzyme-linked immunosorbent assay kits. Results: The overall seropositivity was highest in ORT (45.9%), followed by IBV (37.6%), ART (2.6%), and ILT (0.4%). Out of 360 broiler samples, highest seropositivity was recorded in ORT (43.3%) and lowest in IBV (31.4%). Surprisingly, no broiler samples were found positive for ART and ILT. In case of sonali, the seropositivity was highest in IBV (60%) and lowest in ILT (2%). With respect to types of birds and age groups, the seropositive percentage of all four pathogens was found higher in sonali than broiler. Between two age groups of sonali, the seropositive percentage of ART (12%), ORT (55%), ILT (2%), and IBV (60%) was highest at 2160 weeks of age compared to 520 weeks of age. However, based on location, the seropositive of ORT and IBV was highest in Jamalpur (63.3%) and Fulbariya and Trishal (50%) and lowest in Sreepur (16.7%) and Jamalpur (3.3%). Conclusion: The four pathogens are ubiquitous in nature for the sonali chickens, and the prev¬alence of ORT and IBV was the most prevalent viruses in the study areas. This study indicates a need for improved surveillance and characterization of ORT and ART circulating in all types of poultry in Bangladesh. [J Adv Vet Anim Res 2019; 6(4.000): 561-566]