Haemonchus contortus, an obligatory haematophagus worm infection in small ruminants: Population genetics and genetic diversity.

Haemonchus contortus, a stomach worm, is prevalent in ruminants worldwide. They particularly hamper profitable small ruminant production. Here, we estimate the genetic variation of H. contortus collected from slaughtered goats and sheep from various geographic zones of Bangladesh using multiple genes. To perform this, adult parasites were isolated from the abomasum of slaughtered animals (sheep and goats). Among them, 79 male H. contortus were identified by microscopy. Following the extraction of DNA, ITS-2 and cox1 genes were amplified and subsequently considered for sequencing. After alignment and editing, sequences were analyzed to find out sequence variation, diversity pattern of genes, and population genetics of isolates. Among the sequence data, the analyses identified 19 genotypes of ITS-2 and 77 haplotypes of cox1 genes. The diversity of nucleotides was 0.0103 for ITS-2 and 0.029 for cox1 gene. The dendogram constructed by the genotype and haplotype sequences of H. contortus revealed that two populations were circulating in Bangladesh without any demarcation of host and geographic regions. Analysis of population genetics demonstrated a high flow of genes (89.2 %) within the population of the worm in Bangladesh. The Fst value showed very little amount of genetic difference among the worm populations of Bangladesh but marked genetic variation between different continents. The findings are expected to help explain the risks of anthelmintic resistance and the transmission pattern of the parasite, and also provide a control strategy against H. contortus.

Zoonotic human liver flukes, a type 1 biocarcinogen, in freshwater fishes: genetic analysis and confirmation of molluscan vectors and reservoir hosts in Bangladesh.

BACKGROUND: Opisthorchiid flukes, particularly Opisthorchis viverrini, Opisthorchis felineus, Clonorchis sinensis, and Metorchis spp. are the most common fish-borne zoonotic human liver flukes (hLFs). Liver fluke infections are more prevalent in resource-deprived and underprivileged areas. We herein estimated the prevalence of the metacercariae (MC) of major hLFs in common large freshwater fishes (lFWF) marketed for human consumption from some selected areas of Bangladesh along with detection of their molluscan vectors and reservoirs. METHODS: The current status of fish-borne zoonotic hLF infections in lFWF was investigated along with their molluscan vectors and mammalian reservoir hosts in Mymensingh and Kishoreganj in Bangladesh from July 2018-June 2022 using conventional and multiple molecular techniques, such as PCR, PCR-restriction fragment length polymorphism (RFLP), sequencing, and bioinformatic analyses. The infection rate of fishes was analyzed using the Z-test and the loads of MC were compared using the chi-squared (χ2) test. RESULTS: The MC of C. sinensis, Opisthorchis spp., and Metorchis spp. were detected in 11 species of common and popular lFWF. In lFWF, the estimated prevalence was 18.7% and the mean load was 137.4 ± 149.8 MC per 100 g of fish. The prevalence was the highest (P < 0.05) in spotted snakehead fishes (Channa punctata, 63.6%). The highest rate of infection (P < 0.05) was observed with the MC of C. sinensis (11.8%). Metacercariae were almost equally (P > 0.05) distributed between the head and body of fishes. The infection rate was slightly higher in cultured (19.6%) fishes. The MC of C. sinensis, O. felineus, O. viverrini, and Metorchis orientalis in fishes were confirmed using PCR, PCR-RFLP and bioinformatics. The cercariae of opisthorchiid (Pleurolophocercus cercariae) flukes were only recovered from Bithynia spp. (3.9%, 42 out of 1089). The ova of hLFs from dogs (4.3%, 5 out of 116) and cats (6.0%, 6 out of 100), and adult flukes (M. orientalis) from ducks (41.1% 113 out of 275) were detected. CONCLUSIONS: The MC of hLFs are highly prevalent in fresh water fishes in Bangladesh. Reservoir hosts, such as street dogs, cats, and ducks carried the patent infection, and residents of Bangladesh are at risk.

Comparison of the first time detected Oesophagostomum asperum with Oesophagostomum columbianum in sheep and goats in Bangladesh based on the trinity: Morphology, morphometry and genetic diversity.

Oesophagostomum spp. (Family: Chabertiidae) is keeping a low profile in terms of severity in Bangladesh while maintaining economic loss through disguise within sheep and goats. The study was performed to identify prevalence, confirmation of species through morphology and morphometry followed by phylogeny using ITS2 and COX1 genes. In total 384 slaughterhouse-sourced small and large intestines were pooled from Mymensingh, Kishoreganj, Netrokona, Sherpur and Tangail districts of Mymensingh division. Followed by isolation, O. columbianum and O. asperum were identified following their key morphological features. Notably, O. asperum was first time detected in Bangladesh. The overall prevalence of Oesophagostomum spp. was found 60.93%. The prevalence of O. columbianum (64.95%) was almost double than that of O. asperum (35.04%). Among several characters, only the distance between anus to tail tip showed a significant morphological disparity in female. The Neighbor-joining (NJ) phylogenic trees based on ITS2 and COX1 genes confirmed the study species. The first time identified O. asperum along with morphometry and phylogeny will add value to the fact that nematodes are invisibly present with high prevalence in this country. This study will help to draw specific attention to command a practical control strategy for intervening in economic loss.

Evaluation of accuracies of genomic predictions for body conformation traits in Korean Holstein.

OBJECTIVE: This study aimed to assess the genetic parameters and accuracy of genomic predictions for twenty-four linear body conformation traits and overall conformation scores in Korean Holstein dairy cows. METHODS: A dataset of 2,206 Korean Holsteins was collected, and genotyping was performed using the Illumina Bovine 50K single nucleotide polymorphism (SNP) chip. The traits investigated included body traits (stature, height at front end, chest width, body depth, angularity, body condition score, and locomotion), rump traits (rump angle, rump width, and loin strength), feet and leg traits (rear leg set, rear leg rear view, foot angle, heel depth, and bone quality), udder traits (udder depth, udder texture, udder support, fore udder attachment, front teat placement, front teat length, rear udder height, rear udder width, and rear teat placement), and overall conformation score. Accuracy of genomic predictions was assessed using the single-trait animal model genomic best linear unbiased prediction method implemented in the ASReml-SA v4.2 software. RESULTS: Heritability estimates ranged from 0.10 to 0.50 for body traits, 0.21 to 0.35 for rump traits, 0.13 to 0.29 for feet and leg traits, and 0.05 to 0.46 for udder traits. Rump traits exhibited the highest average heritability (0.29), while feet and leg traits had the lowest estimates (0.21). Accuracy of genomic predictions varied among the twenty-four linear body conformation traits, ranging from 0.26 to 0.49. The heritability and prediction accuracy of genomic estimated breeding value (GEBV) for the overall conformation score were 0.45 and 0.46, respectively. The GEBVs for body conformation traits in Korean Holstein cows had low accuracy, falling below the 50% threshold. CONCLUSION: The limited response to selection for body conformation traits in Korean Holsteins may be attributed to both the low heritability of these traits and the lower accuracy estimates for GEBVs. Further research is needed to enhance the accuracy of GEBVs and improve the selection response for these traits.

Genome-Wide Association Study to Identify QTL for Carcass Traits in Korean Hanwoo Cattle.

This study aimed to identify genetic associations with carcass traits in Hanwoo cattle using a genome-wide association study. A total of 9302 phenotypes were analyzed, and all animals were genotyped using the Illumina Bovine 50K v.3 SNP chip. Heritabilities for carcass weight (CWT), eye muscle area (EMA), backfat thickness (BF), and marbling score (MS) were estimated as 0.42, 0.36, 0.36, and 0.47, respectively, using the GBLUP model, and 0.47, 0.37, 0.36, and 0.42, respectively, using the Bayes B model. We identified 129 common SNPs using DGEBV and 118 common SNPs using GEBV on BTA6, BTA13, and BTA14, suggesting their potential association with the traits of interest. No common SNPs were found between the GBLUP and Bayes B methods when using residuals as a response variable in GWAS. The most promising candidate genes for CWT included SLIT2, PACRGL, KCNIP4, RP1, XKR4, LYN, RPS20, MOS, FAM110B, UBXN2B, CYP7A1, SDCBP, NSMAF, TOX, CA8, LAP3, FAM184B, and NCAPG. For EMA, the genes IBSP, LAP3, FAM184B, LCORL, NCAPG, SLC30A9, and BEND4 demonstrated significance. Similarly, CYP7B1, ARMC1, PDE7A, and CRH were associated with BF, while CTSZ, GNAS, VAPB, and RAB22A were associated with MS. This finding offers valuable insights into genomic regions and molecular mechanisms influencing Hanwoo carcass traits, aiding efficient breeding strategies.

Morphological and phylogenetic analysis of Raillietina spp. in indigenous chickens (Gallus gallus domesticus) in Bangladesh.

Raillietina spp. (Cestoda: Davaineidae), the most common cestodes in indigenous chickens, cause a substantial production loss in poultry industry in Bangladesh. Here, we estimated the prevalence, confirmed the species and determined the genetic pattern of species of Raillietina using molecular tools. We collected and examined 375 chickens randomly from household of different villages of Mymensingh sadar and Gouripur upazila, Mymensingh district and adult parasites were isolated and identified. Genomic DNA was extracted from collected parasites, amplified ITS-2 and ND-1 genes, sequenced and analyzed. Out of 375 samples, 270 (72.0%) were found positive with Raillietina species and mean worm burden was 10.46 ± 0.56. Microscopically, three species of Raillietina, such as R. cesticillus (37.9%), R. echinobothrida (41.1%) and R. tetragona (52.8%) were detected on the basis of their morphological features. The total length, length and width of scolex, sucker and rostellum were also measured. Among different factors, age, farming nature and flock size of chickens were significantly (p < 0.05) influenced Raillietina infections. For further validation, the sequences of ITS-2 gene generated in this study were matched with reference sequences of R. cesticillus, R. echinobothrida and R. tetragona and found 99.63% - 100% similarity. The phylogenetic analyses of ITS-2 and ND-1 sequences were clustered together with the reference sequences of R. cesticillus, R. echinobothrida and R. tetragona confirming microscopic identification. This is the first confirmation of species of Raillietina along with the prevalence of the species, which will be helpful for the formulation of a control strategy and provide basic information for further molecular study.

Genome-Wide Association Studies for Body Conformation Traits in Korean Holstein Population.

The objective of this study was to identify quantitative trait loci (QTL) and nearby candidate genes that influence body conformation traits. Phenotypic data for 24 body conformation traits were collected from a population of 2329 Korean Holstein cattle, and all animals were genotyped using the 50 K Illumina bovine SNP chip. A total of 24 genome-wide significant SNPs associated with 24 body conformation traits were identified by genome-wide association analysis. The selection of the most promising candidate genes was based on gene ontology (GO) terms and the previously identified functions that influence various body conformation traits as determined in our study. These genes include KCNA1, RYBP, PTH1R, TMIE, and GNAI3 for body traits; ANGPT1 for rump traits; MALRD1, INHBA, and HOXA13 for feet and leg traits; and CDK1, RHOBTB1, and SLC17A1 for udder traits, respectively. These findings contribute to our understanding of the genetic basis of body conformation traits in this population and pave the way for future breeding strategies aimed at enhancing desirable traits in dairy cattle.

Ascaridia galli infection in chicken: Pathobiology and immunological orchestra.

BACKGROUND: Ascaridia galli is the largest gut-dwelling helminth of chickens, which confers adverse effects on meat and egg production; thus, on the animal protein supply and the economy. Both adult and immature parasites affect gut health, but larval stages play a major role in pathology. AIMS: Here, we present immunology and pathology of A. galli in chickens. MATERIALS AND METHODS: Literatures were surveyed through online platforms such as PubMed, Google Scholar and Researchgate. RESULTS: The larvae cause excessive mucus production, damage to the intestinal gland, hemorrhage, anemia, diarrhea, and malnutrition. The adult worms can cause death by intestinal obstruction and intussusception. Although both cellular and humoral immunity are involved in fighting against ascariasis, the role of naturally acquired immunity is poorly defined. In cellular immunity, Th-2 cytokines (IL-4, IL-5, IL-9, and IL-13), goblet cells (mucin), gut-associated lymphoid tissues, CD8α+ intraepithelial cells, TCRγδ + T cells, and TGF-ß4 form a protective band. Type 2 immunity provides protection by forming a network of endogenous damage-associated molecular patterns, chitin, and parasitic antigens. Among antibodies, IgY is the most prominent in chickens and provides temporary humoral protection. During parasitic infection, infiltration of various immune cells is evident, especially in the intestinal epithelium, lamina propria, and crypts of the duodenum and jejunum. In chickens older than 12 weeks, gradual reduction of worm burden is more successful than the younger birds. Female chickens exert a short-lived but higher level of protection by passing IgY to chicks in the form of egg yolk antibodies. In laying conditions, immunity differs between breeds. This review provides an overview of the silent but inevitable pathological changes induced by A. galli and the interaction of host immunity with the parasite.

Wide variation of heterozygotic genotypes of recent fasciolid hybrids from livestock in Bangladesh assessed by rDNA internal transcribed spacer region sequencing and cloning.

Fascioliasis causes high economic losses in livestock and underlies public health problems in rural areas, mainly of low-income countries. The increasing animal infection rates in Bangladesh were assessed, by focusing on host species, different parts of the country, and rDNA sequences. Fasciolid flukes were collected from buffaloes, cattle, goats and sheep from many localities to assess prevalences and intensities of infection. The nuclear rDNA internal transcribed spacer (ITS) region including ITS-1 and ITS-2 spacers was analyzed by direct sequencing and cloning, given the detection of intermediate phenotypic forms in Bangladesh. The 35.4% prevalence in goats and 55.5% in buffaloes are the highest recorded in these animals in Bangladesh. In cattle (29.3%) and sheep (26.8%) prevalences are also high for these species. These prevalences are very high when compared to lowlands at similar latitudes in neighboring India. The high prevalences and intensities appear in western Bangladesh where cross-border importation of animals from India occur. The combined haplotype CH3A of Fasciola gigantica widely found in all livestock species throughout Bangladesh fits its historical connections with the western Grand Trunk Road and the eastern Tea-Horse Road. The "pure" F. hepatica sequences only in clones from specimens showing heterozygotic positions indicate recent hybridization events with local "pure" F. gigantica, since concerted evolution did not yet have sufficient time to homogenize the rDNA operon. The detection of up to six different sequences coexisting in the cloned specimens evidences crossbreeding between hybrid parents, indicating repeated, superimposed and rapidly evolving hybridization events. The high proportion of hybrids highlights an increasing animal infection trend and human infection risk, and the need for control measures, mainly concerning goats in household farming management. ITS-1 and ITS-2 markers prove to be useful for detecting recent hybrid fasciolids. The introduction of a Fasciola species with imported livestock into a highly prevalent area of the other Fasciola species may lead to a high nucleotide variation in the species-differing positions in the extremely conserved fasciolid spacers. Results suggest that, in ancient times, frequent crossbreeding inside the same Fasciola species gave rise to the very peculiar characteristics of the present-day nuclear genome of both fasciolids.

Ex-vivo efficacy of commercially available anthelmintics against blood feeding stomach worm Haemonchus contortus of ruminants.

Haemonchus contortus is the most prevalent and pathogenic gastrointestinal nematodes (GINs) in ruminants causing extensive economic losses. It is essential to estimate the efficacy of common commercially available anthelmintics against Haemonchus contortus parasite. Here, we standardized an ex-vivo culture platform for H. contortus and evaluated the efficacy of commonly used anthelmintics namely, albendazole (ABZ), levamisole (LVM), ivermectin (IVM), closantel (CLS) and rafoxanide (RFX). Adult worms were collected from abomasa of slaughtered animals, cultured in MEM, DMEM, M199 or RPMI with or without 20% FBS for up to 72 h. Cultured worms were incubated with ABZ, LVM, IVM, RFX or CLS in DMEM supplemented with 20% FBS at different concentrations (0.5-50 µg/ml) in triplicates and examined at 0, 3, 6, 12, 24, 36 and 48 h post treatment. Of the culture conditions, DMEM supplemented with 20% FBS supported the survival of H. contortus for (P < 0.001) longer period of time which was used in the evaluation of anthelmintics. The efficacy of CLS and RFX were significantly (P < 0.001) higher than other drugs and 100% mortality was observed at 2 µg/ml of CLS and RFX within 12 h post treatment. However, ABZ, LVM, and IVM showed significant effect at the concentration of 50 µg/ml with 48, 36, and 24 h, respectively. Morphological changes included severe cuticle disruption around the buccal cavity, posterior region and vulva as well as loss of cuticle structure integrity coupled with expulsion and fragmentation of digestive components of parasites when treated with 50 µg/ml of ABZ, LVM, and IVM and 2 µg/ml of RFX and CLS. Collectively, DMEM supplemented with 20% FBS can be used as ex-vivo culture platform for maintenance of H. contortus, and RFX and CLS can be used as the promising drugs for the prevention, control and treatment of H. contortus infections.

Diversity and prevalence of gastrointestinal parasites of Black Bengal goats in Natore, Bangladesh.

Objectives: The objective of this work was to estimate the diversity of gastrointestinal (GI) parasite species, their prevalence, and risk factors in Black Bengal goats (BBGs) of Natore, Bangladesh. Materials and Methods: Fecal samples from randomly selected 260 BBGs were processed through Stoll's ova counting method, floatation, and simple sedimentation method. Microscopy-based identification of parasitic eggs, cysts, or oocysts was made. A semi-structured questionnaire-based data on host and management practices were collected from the owner. Data analysis was done using Statistical Package for Social Sciences. Results: The overall prevalence of GI parasites in BBGs was 65.4%, with an individual prevalence of 8.5% for Fasciola gigantica, 21.5% for Paramphistomum spp., 20% for Haemonchus spp., 34.2% for Strongyloides spp., 8.5% for Trichuris spp., and 9.2% for Eimeria spp. No significant effect of host age, gender, body condition, animal rearing system, or housing floor type was observed on parasitism. Animals of young age, female, poorly body-conditioned, living in a free-range system, and housed on a muddy floor had a relatively higher susceptibility to infection. Deworming had a significant impact on reducing the frequency of caprine GI parasitism. Conclusions: Despite the significant effect of anthelmintic, the elevated prevalence of GI parasites in BBGs suggests a critical need for developing effective strategies to prevent caprine parasitoses.

Morphometry and genetic diversity pattern of Cysticercus tenuicollis, an important food-borne taeniid metacestode in goats in Bangladesh.

Cysticercus tenuicollis is a larval stage of Taenia hydatigena resulting in cysticercosis, and responsible for enormous economic loses, especially in livestock production. Here, we planned to determine the prevalence and explore genetic variation of C. tenuicollis isolated from goats based on small subunit ribosomal RNA (rrnS) and cytochrome oxidase subunit 1 (cox1). To do this, samples were collected from different slaughter houses of municipal areas such as Bramhapalli slaughterhouse, Jubileeghat slaughterhouse and Mesuabazar slaughterhouse at Mymensingh sadar, and tentatively identified by morphological and morphometrical analysis. To study genetic variation, DNA was extracted from C. tenuicollis, and amplified rrnS and cox1 genes using specific primers, and were sequenced. Among 1372 examined animals, 177 (12.9%) were infected with C. tenuicollis. Cysts were recovered from peritoneum (7.9%), liver (4.4%) and urinary bladder (0.6%) of the infected animals. Females (18.9%) and adults (20.7%) were significantly more susceptible than male (8.8%) and young (9.3%), respectively. Genetic analysis defined 8 distinct rrnS genotypes and 9 unique cox1 haplotypes among 20 C. tenuicollis isolates. The nucleotide diversities were 0.00283 and 0.00434 for rrnS and cox1 genes, respectively. Neighbor joining (NJ) trees of rrnS and cox1 gene were constructed and the studied sequences were clustered with reference sequences of T. hydatigena with strong nodal support (100%). To compare Bangladeshi isolates, a median joining network was constructed with the population from other geographical regions and hosts. This led to a clustering pattern, but the clusters were not built with unique geographical regions or hosts. In conclusion, this is the first study that describes the genetic variation of T. hydatigena population and suggests the existence of host-specific variants. Therefore, it is fundamental to dispose infected viscera, restrict dog movement and proper management of slaughter house for the prevention and control of cysticercosis.

Efficacy of flukicides on Fasciola gigantica, a food-borne zoonotic helminth affecting livestock in Bangladesh.

Fasciola gigantica, the causative agent of tropical fasciolosis, is a food-borne zoonotic trematode that affects around 80% livestock of Bangladesh. Triclabendazole (TCBZ), nitroxynil (NTON) and oxyclozanide (OCZN) are frequently used against fascioliasis; however, the current status of potency of these flukicides was unknown. In this study, in vitro efficacy of TCBZ, NTON and OCZN at various concentrations on F. gigantica has been evaluated by relative motility (RM), morphological distortions of apical cone through an inverted microscope, architectural and ultra-structural changes through histopathological and scanning electron microscopy (SEM). It is observed that TCBZ, NTON and OCZN at higher concentrations significantly (P < 0.05) reduced RM of the flukes compared to untreated control. NTON at 150 µg mL−1 was the most potent to reduce the motility within 4 h whereas TCBZ and OCZN were much delayed. Histopathological changes showed swollen, extensive cracking, numerous vacuoles and splitting of the tegument surrounding the spines; spine dislodged from its socket in treated flukes compared to untreated worms. Histopathological changes were more conspicuous at higher doses of TCBZ, NTON and OCZN. SEM has shown the disruption of the apical cone, apart from swelling of the tegument on the ventral surface corrugation and disruption of the ventral apical cone. All these changes indicate that NTON is the most potent in killing flukes in vitro among the tested flukicides and suggest the presence of TCBZ-resistant fluke populations in Bangladesh. It is imperative to explore the in vivo effects of these flukicides and subsequently their molecular mechanisms.

Phylogenetic analysis of Eimeria tenella isolated from the litter of different chicken farms in Mymensingh, Bangladesh.

BACKGROUND: Eimeria tenella is the most pathogenic intracellular protozoan parasite of seven Eimeria species causing chicken coccidiosis around the world. This species is particularly responsible for caecal coccidiosis leading to serious morbidity-mortality and financial loss in poultry production. METHODS: The present study explored the genetic diversity of E. tenella. Litter slurry was collected from 18 broiler farms located in Mymensingh district, Bangladesh. Litter samples were processed for oocyst isolation-identification using parasitological techniques followed by genomic DNA extraction from sporulated oocysts. For molecular analysis, the internal transcribed spacer 1 gene of E. tenella was amplified using species-specific primers and sequenced. After editing and alignment, 263 bp sequences were used for analysis. RESULTS: Genetic analysis showed seven distinct genotypes and detected six single nucleotide polymorphisms among the 18 E. tenella isolates. The nucleotide and genotype diversity were 0.00507 and 0.8235, respectively. A phylogenetic tree was constructed with 66 sequences (seven studied genotypes and 59 reference sequences from GenBank database). The neighbour-joining tree represented that the studied E. tenella isolates were grouped with reference E. tenella isolates with strong nodal support (100%) and the nucleotide sequences of E. tenella, E. necatrix, E. acervulina, E. brunetti, E. maxima, E. mitis and E. praecox formed separate clusters without any geographical boundaries. CONCLUSIONS: This is the first study on the genetic analysis of E. tenella from Mymensingh district, Bangladesh. These findings will provide baseline data on the species conformation and genetic variations of E. tenella. Further extensive investigation will be needed to reveal the population genetic structure of this parasite and thus will facilitate the planning of effective control strategies.

A large-scale epidemiological investigation on trematode infections in small ruminants in Bangladesh.

BACKGROUND: The important trematode species in small ruminants: Paramphistomum sp., Fasciola spp. and Schistosoma spp. seriously affect the productivity of domestic ruminants in endemic areas. METHODS: In the present study, we identified the potential risk factors associated with trematodes infections in small ruminants in seven topographic zones of Bangladesh using simple sedimentation and modified Stoll' ova counting technique. RESULTS: A total of 2440 samples were examined, where 965 were found positive with one or more trematode species with an overall prevalence of 39.5% (95% CI, 37.6%-41.5%) and intensity of infection was 264.77 ± 9.86 egg per gram of faeces. Three trematode species were identified namely Paramphistomum sp. (34.1%, 32.2%-36.0%), Fasciola spp. (7.5%, 6.5%-8.6%) and Schistosoma spp. (2.7%, 2.1%-3.5%). Prevalence of co-infection was 4.8%. The spatial distribution of trematode infections varied from 29.5% to 53.6%. Univariate analysis revealed that physiological condition of females, body condition, farming system, deworming and season were significantly (p < 0.05) associated with trematodes infections in small ruminants. By multiple logistic regression model, three factors such as physiological condition of females (pregnant and lactating), poor body condition and animals without deworming were identified as potential risk factors for trematodes infection in small ruminants. CONCLUSION: Trematode infections are prevalent in the study areas and Paramphistomum sp. is most common in different areas among the identified trematodes species. Government should take necessary action to appraise an effective control strategy of trematode infections in small ruminants.

Research Note: Genetic analysis, pathology, and vectors of echinostomiasis, a zoonotic helminth infection in chickens in Bangladesh.

Echinostomes (Trematoda: Echinostomatidae) are food-borne zoonotic flatworms that affect birds, animals and humans, and has been classified as neglected tropical diseases (NTDs) by the World Health Organization (WHO), which cause severe enteritis in poultry and hamper production. Here, we confirmed the species of echinostomes affecting chickens in Bangladesh along with their genetic analyses, pathology and vectors. We isolated and identified adult worms from chickens, cercariae from fresh water snails and metacerariae (MC) from some wild fishes. We recovered Echinostoma revolutum (10.3%) and Hypoderaeum conoideum (6.0%) from chickens. Zoonotic E. revolutum was confirmed by amplifying nad1 gene and subsequent sequencing. Several mutations were detected in nad1 gene and our isolates belonged to the Euro-Asian clade. We observed thickening of mucosal layer, hyperplasia of goblet cells, infiltration of eosinophils, lymphocytes and must cells in the infected intestine. About 5.3% snails were infected and the highest percentage of infection was found in Lymnaea luteola (12.1%). Echinostome infection in snails was the highest in November (9.6%) and lowest in February (3.1%) in Bangladesh. MC of echinostomes were identified from blue panchax (Aplocheilus panchax) and tank goby (Glossogobius giuris). In conclusion, echinostomiasis is a notable big problem in indigenous chickens in Bangladesh and people, especially, villagers are at risk.

Frequency of benzimidazole resistance in Haemonchus contortus populations isolated from sheep and goats in Bangladesh.

Anthelmintic resistance against gastrointestinal nematodes especially H. contortus of sheep and goat is a global issue. To address the gravity and extension of AR in Bangladesh, genotyping of 160 adult H. contortus parasitesw ere performed to confirm benzimidazole resistance allele from different geographic zones of Bangladesh based on allele specific PCR (AS-PCR). The genotype frequencies were 9.4% for homozygous resistant (rr), 61.2% for heterozygous (rS) and 29.4% for homozygous susceptible (SS) among the selected areas. The allelic frequency of the mutation conferring resistance (r) ranged from 27.5% to 52.5% indicating substantial existence of benzimidazole resistance in H. contortus in small ruminant nematodes. Therefore, it can be concluded that genotyping the F200Y polymorphism can be used to monitor the resistance and thereby to enhance the control on the development of anthelmintic resistance against H. contortus in small ruminant nematodes.

ITS1-PCR based identification of chicken Eimeria species in poultry litter from Mymensingh district, Bangladesh.

OBJECTIVES: The purpose of this study was to determine the species composition of Eimeria circulating in Mymensingh district, Bangladesh, using Internal Transcribed Spacer 1 (ITS1) sequences in polymerase chain reaction (PCR) assay. MATERIALS AND METHODS: Coccidian oocysts were isolated and sporulated in a solution containing 2% potassium dichromate from litter slurry collected from 13 commercially active broiler farms in the research region. Genomic DNA was isolated from sporulated oocysts and used to amplify the Eimeria species-specific ITS1 gene by PCR amplification. Electrophoresis of 1.5% agarose gel was used to visualize the amplified PCR products. RESULTS: In the study samples from Mymensingh district, Bangladesh, the presence of Eimeria brunetti, Eimeria acervulina, Eimeria necatrix, Eimeria mitis, and Eimeria tenella was identified. CONCLUSIONS: The findings of this study may shed light on the zonal approach to chicken coccidiosis control. Additionally, it suggests that ITS1-based PCR might be used in the field to accurately identify Eimeria species.

Prevalence and factors influencing gastrointestinal parasitic infections in sheep in Bangladesh.

Gastrointestinal (GI) parasites are major contributors to decrease productivity in livestock over the world. A cross-sectional study was conducted in different areas of Bangladesh to determine the prevalence of GI parasitic infections and their association with the biotic and abiotic factors in sheep. A total of 572 faecal samples were collected from the selected areas of Bangladesh and microscopic examination was performed for the identification of parasites using flotation and sedimentation technique. Out of 572, 441 animals were found infected with one or more species of GI parasites with an overall prevalence of 77.1%. Nine types of parasites from four different classes were detected namely Strongyles (42.1%), Strongyloides sp. (27.1%) and Trichuris sp. (1.0%), Moniezia sp. (2.4%), Paramphistomum cervi (32.5%), Fasciola gigantica (6.1%) and Schistosoma sp. (3.5%), coccidia (16.6%) and Balantidium coli (7.9%). Nematodes infections (56.8%) were significantly highest among trematodes (37.9%), protozoa (24.4%) and cestode (2.4%). In the present study, all the biotic factors including sex, age, physiological condition of female and body condition score (BCS) of animals were insignificantly (p>0.05) associated with the prevalence of GI parasitic infection in sheep but among the abiotic factors, muddy housing of animals, rainy season, having no knowledge about GI parasites and illiteracy of farmers were significantly (p<0.05) associated with the GI parasitic infections. This epidemiological investigation will assist to build a suitable control program against GI parasites in sheep and thus, help to prevent production loss and increase livelihood of small holder farmers.

Small-scale farmers' perception and practice on coccidiosis management in broiler farm at Gazipur, Bangladesh.

Farmers' knowledge and farm management practices can potentially help to mitigate the prevalence and severity of chicken coccidiosis, a devastating protozoan disease. Here, we assessed the farmers' perception and on-farm practices of coccidiosis management in small-scale broiler farms at Gazipur district, Bangladesh. A semi-structured questionnaire was used to obtain data from the randomly selected 119 small-scale broiler producers followed by detection of coccidiosis cases through gross and microscopic examination of 58 broilers. Overall bird-level prevalence of coccidiosis was 34.48% in this study. Survey data revealed that most of the farmers maintained all-in-all-out strategy (68.91%), good quality chicks (73.11%) and floor system rearing (96.63%) in their farms, and a mixture of old and new litter was used as bedding in all the study farms. Interval between flocks in 57.99% of farms was 8 to 14 days. Prevalence of coccidiosis was recorded in all surveyed farms, even though routine anticoccidial treatment. At the age of 15 to 18 days of broiler, 68.07% of farmers used chemoprophylaxis. Usage of amprolium was the highest (74.78%) followed by toltrazuril (55.46%), sulphaquinoxaline (23.52%), sulphaclozine (28.57%), sulphadimidine (24.36%) and sulphadimethoxine (24.36%). Traditional herbal medicines like extracts of bollygum (Litsea glutinosa), sal (Shorea robusta) and arjuna (Terminalia arjuna) bark and leaves were also in-use by 4.20% farmers during coccidiosis outbreak. In conclusions, this study explored the field scenario of coccidiosis, relevant farmers' knowledge and practices, and recommends the judicious use of drugs for safe broiler production.