Morphometry and Molecular Identification of Haemonchus Cobb, 1898 (Trichostrongylidae: Nematoda) Isolates from Small Ruminants in Tanzania Based on Mitochondrial cox 1 and rRNA-ITS genes.

The genus Haemonchus is the major abomasal parasite of ruminants responsible for substantial economic losses in tropical and temperate regions. This study was conducted to clarify the morphometry and molecular characterisation of Haemonchus species isolated from sheep in Babati district, Tanzania. A total of 486 trichostrongylid nematodes were recovered from five sheep. Of the total worms, 106 nematodes were distinguished by 37 males and 69 females. The asymmetrical length of dorsal ray and the distance of bulb at the apex of spicules were used for identification of males. In females, the linguiform vulvar flap was the most predominant with 33 out of 69 (48%) compared with knobbed morph type which was 25/69 (36%) and smooth morph type with 11/69 (16%). Partial cox1 sequence fragments of Haemonchus contortus isolates showed 98.8%, 99.3%, 99.7%, 99.5%, 99.3%, and 98.4% in male, smooth, knobbed, linguiform A, linguiform B, and linguiform C, respectively; with the average nucleotide divergence ranged from 1.03 to 2.35%. The amplified fragments of ITS-2 genes in knobbed, linguiform A, and smooth morphotypes revealed 99.4%, 98.5%, and 98.3%, respectively. Phylogenetic analysis was evaluated by employing Bayesian inference and maximum-likelihood, and the tree was distinctly separated into three clusters focusing on H. contortus in cluster I within the family Haemonchidae. Genetic drifting, mutation, and modification of the morphological features of the Haemonchus species described to have an impact on the development of drug resistance. Species identification is necessary to understand which species infect animal host. We recommend more studies on the parasites intensity and the strategies for controlling Haemonchus species in Tanzania.

Insights to helminth infections in food and companion animals in Bangladesh: Occurrence and risk profiling.

INTRODUCTION: A better understanding of the epidemiology of helminths in animal hosts is important in order to ensure animal welfare, public health and food safety. The aim of this study was to explore parasitism in common animals in Bangladesh. Perception and understanding of animal owners regarding parasitic diseases management were also assessed. MATERIALS AND METHODS: A total of 550 fecal samples were examined from common animals (cattle, goat, pig, chicken, dog, and cat) across three different areas of Bangladesh (Dhaka, Sylhet, and Chattogram) from January 2020 to March 2021. Associated risk factors were assessed through questionnaire surveys among 50 animal owners. Parasitological assessment was done using the combined sedimentation-flotation method, and factors associated with infection were modeled using mixed-effects logistic regression. RESULTS: Helminths including Toxocara sp., Spirometra sp., Capillaria sp., Trichuris sp., opisthorchiid, Ascaris suum, Fasciola sp., Paramphistomum sp., strongyles, hookworms, roundworms, taeniid, and acanthocephalans were detected in the examined animals, and overall prevalence was 59.3% (95% CI = 54.1-62.8). Parasites were found in 61.3% (245/400) of food animals and 54.0% (81/150) of companion animals. Animal owners have a good understanding of parasite infections; however, that knowledge was not being translated into practice. Logistic regression analysis showed that frequency of deworming, animal husbandry practice, contact with untreated animals, and treatment-seeking behaviors were significantly associated with parasitic infection. CONCLUSION: Several types of zoonotic parasites are widely prevalent in animals of Bangladesh and pose a potential risk to human health. This study highlights the need to diagnose animal parasitic infection and intensified case management to avoid spillovers to animals and humans.

Genotypes of Echinococcus Species from Cattle in Tanzania.

Cystic echinococcosis is a zoonotic parasitic disease caused by Echinococcus species. Tanzania is one of the endemic countries with cystic echinococcosis. This study focussed on identifying genotypes of Echinococcus spp. in Tanzania. We collected 7 cysts from cattle in Mwanza municipal (n=4) and Loliondo district (n=3). The cysts from Mwanza were all E. ortleppi and fertile. In contrast, the cysts from Loliondo were all E. granulosus sensu stricto and sterile. Two from the 4 cysts were a new haplotype of E. ortleppi (G5). These results can improve the preventive and control programs for humans and livestock in Tanzania. To our knowledge, this study is considered the first to identify the genotype and haplotype of Echinococcus spp. in Tanzania.

Insights into geohelminth contamination in Bangladesh: feasibility of a modified diagnostic method and prevalence study.

INTRODUCTION: The objectives of this study were to evaluate a modified method for isolating geohelminth eggs and to assess the geohelminth contamination in Bangladesh. METHODS: The efficacy of the modified method was evaluated using naturally contaminated and experimentally seeded soil samples. In total, 240 samples were assessed from four different sites in three geographic contexts. A questionnaire survey with 50 professionals was conducted. RESULTS: The modified method showed considerable efficacy in isolating parasitic eggs from naturally contaminated soil (54.0%) and experimentally seeded soils (63.0% for Toxocara eggs and 52.0% for Ascaris eggs). The modified method was described as convenient by the majority of participants. Overall prevalence was 52.5%, with several species of helminth observed, including Toxocara sp., Ascaridia galli/Heterakis gallinarum, Ascaris sp., hookworms/strongyles, Capillaria sp., Trichuris sp., and taeniids). The contamination rate was found to be higher around livestock farms (76.7%), followed by latrines (63.3%), households (41.6%), and schools (28.3%). CONCLUSIONS: The modified method was shown to be feasible in terms of field applicability and egg recovery rate, and could be adopted in low-resource settings. A substantial prevalence of geohelminths was observed, with some of the species associated with zoonoses. These findings highlight the urgent need for widespread mapping of geohelminths to avoid spillovers to animals and humans.

Mitochondrial Genome of Spirometra theileri Compared with Other Spirometra Species.

This study was carried out to provide information on the taxonomic classification and analysis of mitochondrial genomes of Spirometra theileri. One strobila of S. theileri was collected from the intestine of an African leopard (Panthera pardus) in the Maswa Game Reserve, Tanzania. The complete mtDNA sequence of S. theileri was 13,685 bp encoding 36 genes including 12 protein genes, 22 tRNAs and 2 rRNAs with absence of atp8. Divergences of 12 protein-coding genes were as follow: 14.9% between S. theileri and S. erinaceieuropaei, 14.7% between S. theileri and S. decipiens, and 14.5% between S. theileri with S. ranarum. Divergences of 12 proteins of S. theileri and S. erinaceieuropaei ranged from 2.3% in cox1 to 15.7% in nad5, while S. theileri varied from S. decipiens and S. ranarum by 1.3% in cox1 to 15.7% in nad3. Phylogenetic relationship of S. theileri with eucestodes inferred using the maximum likelihood and Bayesian inferences exhibited identical tree topologies. A clade composed of S. decipiens and S. ranarum formed a sister species to S. erinaceieuropaei, and S. theileri formed a sister species to all species in this clade. Within the diphyllobothridean clade, Dibothriocephalus, Diphyllobothrium and Spirometra formed a monophyletic group, and sister genera were well supported.

Morphometrical and Molecular Characterization of Oesophagostomum columbianum (Chabertiidae: Oesophagostominae) and Haemonchus contortus (Trichostrongylidae: Haemonchinae) Isolated from Goat (Capra hircus) in Sylhet, Bangladesh.

This study was aimed at describing two (2) intestinal nematodes from naturally infected native breed of goats (Capra hircus) in Bangladesh, identified as Oesophagostomum columbianum (Curtice, 1890) Stossich 1899 and Haemonchus contortus (Rudolphi, 1803) Cobb, 1898. The identification was made based on morphometric features and was confirmed by amplifying internal transcribed spacer (ITS) and cytochrome c oxidase (cox1) gene. Well-developed lateral alae, distinct cervical papillae anteriorly to esophageal expansion, and male spicule length (0.73-0.79 mm, n = 2) were characteristically observed in O. columbianum. At the same time, male spicule length (0.40-0.46 mm, n = 2) and position of female vulvar flap (4.30-4.54 mm from posterior end, n = 3) were observed in H. contortus. DNA sequence homology of the ITS and cox1 gene of both specimens revealed the same results, showing similarity to the GenBank sequences of O. columbianum (GenBank No. KC715827; JX188470) and H. contortus (GenBank No. KJ724377; HQ389229). Phylogenetic analysis computed by maximum livelihood (ML) from the ITS nucleotide sequences revealed that the O. columbianum and H. contortus isolates identified in this study were clustered in the same clade with isolates from China and Iran, respectively. This study, for the first time, illustrates the characteristics of O. columbianum and H. contortus in Bangladesh, combining both morphological and molecular data. The universal primer-based polymerase chain reaction (PCR) protocol could be an economical and efficient option for researchers from poor resource settings for precise identification of nematodes. The information generated in this study may contribute to formulating effective control strategies against these nematodes.

Insight into One Health Approach: Endoparasite Infections in Captive Wildlife in Bangladesh.

Introduction: Endoparasites in captive wildlife might pose a threat to public health; however, very few studies have been conducted on this issue, and much remains to be learned, especially in limited-resource settings. This study aimed to investigate endoparasites of captive wildlife in Bangladesh. Perception and understanding of veterinarians regarding one health and zoonoses were also assessed. Materials and Methods: A cross-sectional study was conducted from October 2019 to August 2020. A total of 45 fecal samples from 18 different species of wild animals (i.e., 11 species of mammals: herbivores, carnivores, and omnivores, six birds, and a single reptile species) were collected randomly. Parasitological assessments were done by modified formalin ether sedimentation technique and rechecked by Sheather's sugar floatation technique. Molecular identification of Spirometra spp. was conducted by amplifying the cytochrome c oxidase 1 (cox1) gene. Questionnaire surveys among 15 veterinarians and an in-depth interview (IDI) with a zoo officer were conducted. Results: Helminths (Spirometra sp., Capillaria sp., Ascaridia/Heterakis, opisthorchiid, strongyles, acuariid, hookworms, roundworms, and unidentified nematode larvae) and protozoa (coccidian oocyst) were identified, and the overall prevalence was 48.9% (22/45). The cox1 sequences (341 bp) of the Bangladesh-origin Spirometra species from lion showed 99.3-99.7% similarity to the reference sequences of Spirometra decipiens (GenBank No: KJ599679.1; MT122766). The majority of study participants (86.6%) agreed about the importance of endoparasite control in zoo animals, and 73.3% expressed that the one health concept should be promoted in Bangladesh. Only 6.7% of veterinarians perceived confidence in diagnosing parasitic diseases and preventing antiparasiticidal resistance. Conclusions: In the present survey, we found a considerable prevalence of endoparasites in captive wildlife. For the first time, zoonotically important S. decipiens from lion was molecularly characterized in Bangladesh. Veterinarian training is required to improve parasite control knowledge and practice. This study highlights the need for routine parasitological assessment, promotion of one health, and improvement of the implementation of current parasite control strategies in zoo animals.

First Infestation Case of Sarcoptic Mange from a Pet Rabbit Oryctolagus Cuniculus in Republic of Korea.

In November 2019 a 5-month-old mixed-breed rabbit presented to Chungbuk National University Veterinary Teaching Hospital, Cheongju-si, Chungbuk, Republic of Korea (Korea) with symptoms comprising pruritus, crusts on skin, poor appetite and reduced defecation. The rabbit was purchased 2 months prior from a pet shop located in a big market, and that the symptoms were first observed about 2 weeks prior to the hospital visit. Physical examination revealed that the patient had crust formation and alopecia on the nose together with lesions on the digits. A skin scraping test was performed using mineral oil and a high density of mites was observed by microscopy. Each mite showed a round, tortoise-like body with 4 comparatively short pairs of legs. The anus was located at the terminal unlike with suspected pathogen, Notoedres cati. Based on morphological characteristics, we identified the mite as Sarcoptes sp. Ivermectin was administered weekly by subcutaneous injection at a dosage of 0.4 mg/kg, and 4 weeks of follow-up study revealed the patient was fully recovered. And no more mites were detected from the case. This is the first case report of sarcoptic mange in a pet rabbit in Korea.

Morphological and Molecular Identification of Spirometra Tapeworms (Cestoda: Diphyllobothriidae) from Carnivorous Mammals in the Serengeti and Selous Ecosystems of Tanzania.

Spirometra tapeworms (Cestoda: Diphyllobothriidae) collected from carnivorous mammals in Tanzania were identified by the DNA sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and internal transcribed spacer 1 (ITS1), and by morphological characteristics. A total of 15 adult worms were collected from stool samples and carcasses of Panthera leo, Panthera pardus, and Crocuta crocuta in the Serengeti and Selous ecosystems of Tanzania. Three Spirometra species: S. theileri, S. ranarum and S. erinaceieuropaei were identified based on morphological features. Partial cox1 sequences (400 bp) of 10 specimens were revealed. Eight specimens showed 99.5% similarity with Spirometra theileri (MK955901), 1 specimen showed 99.5% similarity with the Korean S. erinaceieuropaei and 1 specimen had 99.5% similarity with Myanmar S. ranarum. Sequence homology estimates for the ITS1 region of S. theileri were 89.8% with S. erinaceieuropaei, 82.5% with S. decipiens, and 78.3% with S. ranarum; and 94.4% homology was observed between S. decipiens and S. ranarum. Phylogenetic analyses were performed with 4 species of Spirometra and 2 species of Dibothriocephalus (=Diphyllobothrium). By both ML and BI methods, cox1 and ITS1 gave well supported, congruent trees topology of S. erinaceieuropaei and S. theileri with S. decipiens and S. ranarum forming a clade. The Dibothriocephalus species were sisters of each other and collectively forming successive outgroups. Our findings confirmed that 3 Spirometra species (S. theileri, S. ranarum, and S. erinaceieuropaei) are distributed in the Serengeti and Selous ecosystems of Tanzania.

Identity of Spirometra theileri from a Leopard (Panthera pardus) and Spotted Hyena (Crocuta crocuta) in Tanzania.

In the present study, a Spirometra species of Tanzania origin obtained from an African leopard (Panthera pardus) and spotted hyena (Crocuta crocuta) was identified based on molecular analysis of cytochrome c oxidase I (cox1) and NADH dehydrogenase subunit I (nad1) as well as by morphological observations of an adult tapeworm. One strobila and several segments of a Spirometra species were obtained from the intestine of an African male leopard (Panthera pardus) and spotted hyena (Crocuta Crocuta) in the Maswa Game Reserve of Tanzania. The morphological characteristics of S. theileri observed comprised 3 uterine loops on one side and 4 on the other side of the mid-line, a uterine pore situated posterior to the vagina and alternating irregularly either to the right or left of the latter, and vesicular seminis that were much smaller than other Spirometra species. Sequence differences in the cox1 and nad1 genes between S. theileri (Tanzania origin) and S. erinaceieuropaei were 10.1% (cox1) and 12.0% (nad1), while those of S. decipiens and S. ranarum were 9.6%, 9.8% (cox1) and 13.0%, 12.6% (nad1), respectively. The morphological features of the Tanzania-origin Spirometra specimens coincided with those of S. theileri, and the molecular data was also consistent with that of S. theileri, thereby demonstrating the distribution of S. theileri in Tanzania. This places the leopard (Panthera pardus) and spotted hyena (Crocuta Crocuta) as new definitive hosts of this spirometrid tapeworm.