An insight into the epidemiology of foodborne zoonotic fascioliasis in small ruminants in northwestern region of Bangladesh.

Fascioliasis is one of the most common foodborne zoonotic infection of ruminants in Bangladesh. To estimate the prevalence and associated risk factors of fascioliasis and extent of liver damage, 825 livers of sheep and goats were randomly inspected during onsite slaughterhouse visiting in Naogaon, Natore, Rajshahi and Joypurhat districts. The overall prevalence of fascioliasis was 25.09% and significantly (P = 0.008) higher in goats (26.11%) than sheep (24.00%). During gross inspection, Fasciola infected livers were increased in size, fibrosed, fatty, multiple white or reddish necrotic foci on the parietal surface, hard to cut, calcified, and numerous mature and immature flukes were also observed. In histoarchitecture, inflammatory cell infiltration in the hepatic parenchyma and periportal area, fibrous connective tissue proliferation around necrotic area, hyperplastic bile duct, congestion, and primary biliary cirrhosis were seen in acute and chronic fascioliasis. Epidemiological investigations revealed that fascioliasis was higher in goats than sheep. Age, sex, BCS and season were found to have statistically significant associations with fascioliasis in goats. In case of sheep, age (OR = 5.8671; 95% CI: 2.9482-11.6757, P < 0.0001), sex (OR = 3.7317; 95% CI: 1.9052-7.3094, p < 0.0001), BCS (OR = 6.0346; 95% CI: 1.7986-20.2472, p < .0001), and season (OR = 8.2308; 95% CI: 3.9922-16.9693, p = < .0001) were also found to have statistically significant associations with fascioliasis. Results of the study can help for molecular epidemiology of fascioliasis in small ruminants to plan fluke control programs for safe food production.

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.

Molecular and phylogenetic characterization of zoonotic Trichostrongylus species from goats for the first time in Bangladesh.

BACKGROUND: Trichostrongylus is one of the most important nematodes infecting animals and humans. The current study was designed to identify the Trichostrongylus species infecting goats by multiplex PCR and phylogenetic analysis. METHODS: A total of 124 goats' viscera were collected from different abattoirs of Mymensingh division. Trichostrongylus species were isolated and characterized based on morphometry, multiplex PCR and phylogenetic analysis. RESULTS: Among 124 viscera of goats, 39 were positive with two species, Trichostrongylus colubriformis and Trichostrongylus vitrinus, revealing an overall 31.45% prevalence. Morphological identification of Trichostrongylus species was confirmed by multiplex PCR amplification of the ITS2 gene and sequencing. Partial sequencing of the ITS2 gene of two species revealed seven single nucleotide polymorphisms (three transitions and four transversions) in this study. The neighbor-joining phylogenetic tree demonstrated that T. colubriformis and T. vitrinus isolates were clustered together with the reference sequences that belong to the clade A and B without any geographical boundaries. CONCLUSIONS: This is the first report on molecular and phylogenetic analysis of Trichostrongylus species from ruminants in Bangladesh. These results provide the baseline data for understanding the zoonosis and epidemiology of this parasite in Bangladesh and global perspectives.

Efficacy of clinically used anthelmintics against toxocariasis of buffalo calves in Bangladesh.

Toxocariasis is a paramount parasitic disease having > 50% prevalence among newborn buffalo calves in Bangladesh. The objective of this study was to compare the efficacy of clinically used anthelmintics and their subsequent effect on the haematological parameters and body weight in buffalo calves in commercial buffalo farms in coastal region. Thirty-two buffalo calves below 3 months of age with clinical Toxocara infection had been selected and treated with albendazole (ABZ), levamisole (LVM) and ivermectin (IVM). The EPG counts, hematological parameters and initial body weight of buffalo calves were recorded on the day of anthelmintic treatment (day 0). Fecal samples were collected on day 14 and 28 including hematological parameters and body weight were recorded on day 28 post-treatment. The efficacy (%) of anthelmintics were estimated by fecal egg count reduction test (FECRT). The parameters like Hb, PCV, ESR, TEC, TLC were analyzed from the blood samples. The FECRT revealed 96.83% efficacy for IVM followed by 94.23% and 85.84% for LVM and ABZ, respectively. Anthelmintic treated calves showed significant (p < 0.01) increase in Hb, PCV, TEC and body weight, and decrease in ESR and TLC as a result of worm expulsion from buffalo calves after 28 days of post-treatment. Among the tested anthelmintics, IVM was found to be more effective against toxocariasis in buffalo calves. This is a novel information on anthelmintics efficacy in buffalo calves in Bangladesh. Details study is recommended on the efficacy of anthelmintics in different buffalo management systems by in vitro egg hatch assay (EHA) test.

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.

Prevalence of hookworm infections among stray dogs and molecular identification of hookworm species for the first time in Bangladesh.

Hookworms are the most common and voracious blood-sucking parasites of the small intestines of mammalian hosts such as dogs, cats, ruminants and humans. Canine hookworms are endemic in the Southeast Asian countries including Bangladesh. There is scarcity of information on the prevalence of hookworms of stray dogs in Bangladesh. The present study determined the prevalence of canine hookworms using fecal examination followed by morphometric and molecular identification. Fecal samples were collected from 320 stray dogs living in rural areas of Mymensingh district (Gauripur upazila, Mymensingh sadar upazila and Tarakanda upazila) and hookworm eggs were identified using the flotation techniques. The overall prevalence of hookworm was 79.1% through microscopic examination. Estimated fecal prevalence was higher in Gauripur upazila (89.7%) followed by Mymensingh sadar upazila (84.8%) and Tarakanda upazila (53.2%). Five hookworm species were identified based on the morphometric examination, namely, Ancylostoma caninum, Ancylostoma ceylanicum, Ancylostoma tubaeforme, Ancylostoma braziliense and Ancylostoma duodenale, respectively. Polymerase Chain Reaction (PCR) was performed with the genomic DNA by targeting the 5.8S rRNA (~ 404 bp) and Cytochrome oxidase-1 (Cox 1, ~ 450 bp) and confirmed the identification for the first time in Bangladesh. This study reveals that stray dogs may act as reservoir hosts of human hookworm infection. Further detail molecular study is warranted to explore the genetic diversity of hookworms that infect both dogs and human in Bangladesh.

Prevalence and risk factors of Toxocara vitulorum infection in buffalo calves in coastal, northeastern and northwestern regions of Bangladesh.

Buffalo (Bubalus bubalis) is popularly known as the black gold of South Asia, consisting of 97% of the world buffalo population. Among the parasitic infections, Toxocara vitulorum is one of the most common and harmful parasites of buffalo calves in Bangladesh. A cross-sectional study was conducted to explore the prevalence and associated risk factors of T. vitulorum infection of buffalo calves in four regions of Bangladesh. A total of 1751 fecal samples were collected and examined using flotation followed by the McMaster technique for counting the eggs per gram of feces (EPG) of T. vitulorum. The overall prevalence of T. vitulorum infection in buffalo calves was 22.9%. Significantly (p < 0.001) higher prevalence was found in the Barishal coastal area (35.7%) followed by Chattogram coastal area (29.2%), northeastern region (15.5%) and northwestern region (8.3%). Buffalo calves aged 1-3 months were heavily infected with T. vitulorum (51.7%) which was statistically different (p < 0.001) compared to those >3-6 months (27.6%) and > 6-12 months (6.5%). According to univariate analysis, coastal regions, rainy season, young age, gender, indigenous river type, buffalo calves with poor body condition and soft feces were found significantly associated with T. vitulorum infections. Coastal regions, rainy season and young age were identified as the risk factors of T. vitulorum infection in buffalo calves by final logistic regression model. This study reveals that T. vitulorum infection is endemic in Bangladesh and widely distributed in the coastal regions. Therefore, attention to buffalo calves regarding in-depth clinical effects and current therapeutic approaches against this nematode should be evaluated along with the economic impact of infection. Exploring the genetic diversity of T. vitulorum may help to reveal the host-parasite relationship in the future.

Morphological and phylogenetic analyses of Rhipicephalus microplus ticks from Bangladesh, Pakistan and Myanmar.

Ticks of the Boophilus subgenus, classified in the genus Rhipicephalus, are widespread in subtropical and tropical regions, but knowledge on their distribution in Asia is fragmentary. The most important representatives belong to the Rhipicephalus (Boophilus) microplus complex, which is composed of species that share many morphological similarities and are therefore difficult to distinguish. In this study, adult Rhipicephalus (Boophilus) ticks from Bangladesh, Myanmar and Pakistan were subjected to phylogenetic and Scanning Electron Microscopy (SEM) analyses. Phylogenetic analyses of cytochrome oxidase 1 (cox1), internal transcribed spacer 2 (ITS2) and 12S rRNA gene sequences confirmed that the R. microplus complex consists of at least five taxa: R. annulatus, R. australis, and R. microplus clades A-C. Ticks from Bangladesh, Myanmar and Pakistan were assigned to R. microplus clade C. SEM images revealed a wide range of variation in the morphology of the clade C adults, including morphological features previously identified as critical for distinguishing R. microplus from R. australis, which is illustrative for the complications in identifying species within the R. microplus complex using morphology only. Further morphological, genomic and crossbreeding studies are required to elucidate the species status of R. microplus clades A-C.