Metabarcoding of pathogenic parasites based on copro-DNA analysis of wild animals in South Korea.

Four species of dominant wild animals, namely, Prionailurus bengalensis euptilurus, Nyctereutes procyonoides koreensis, Hydropotes inermis argyropus, and Sus scrofa coreanus, are hosts of potential infectious agents, including helminths and protozoa. Therefore, it is necessary to analyze the infectious agents present in these wild animals to monitor and control the spread of pathogens. In the present study, fecal samples from 51 wild animals were collected from the mountains of Yangpyeong, Hoengseong, and Cheongyang in South Korea and metabarcoding of the V9 region of the 18S rRNA gene was performed to identify various parasite species that infect these wild animals. Genes from nematodes, such as Metastrongylus sp., Strongyloides spp., Ancylostoma sp., and Toxocara sp., were detected in the fecal samples from wild animals. In addition, platyhelminthes, including Spirometra sp., Echinostomatidae gen. sp., Alaria sp., Neodiplostomum sp., and Clonorchis sp., and protozoa, including Entamoeba sp., Blastocystis sp., Isospora sp., Tritrichomonas sp., Pentatrichomonas sp., and Cryptosporidium sp., were detected. In the present study, various parasites infecting wild animals were successfully identified using metabarcoding. Our technique may play a crucial role in monitoring parasites within wild animals, especially those causing zoonoses.

Adhesive Bowel Obstruction (ABO) in a Stranded Narrow-Ridged Finless Porpoise (Neophocaena asiaeorientalis sunameri).

In this case report, we present a rare occurrence of a narrow-ridged finless porpoise (Neophocaena asiaeorientalis sunameri), discovered on the coast of Jeju Island, Republic of Korea, that was afflicted with adhesive bowel obstruction (ABO), a life-threatening condition that has scarcely been reported in cetaceans. Diagnosis of ABO was confirmed via radiological and clinical assessments. Post-mortem computed tomography and necropsy revealed ABO between two loops of the jejunum at the L8 level. The mesenteric tissue covering the intestinal lesion was severely thickened with increased tension. Both bowel loops were fixed to the mesentery and acutely angulated, leading to asymmetrical thickening of the cross-sectional bowel walls. The intestinal lumen was stenosed because of pressure from the firm mesenteric band, and no fecal matter was observed in the lumen of the posterior bowel or rectum. Calcified nodules were detected, and histological analysis suggested parasitic or suspected post-parasitic infections. The primary cause of the intestinal lesions is presumed to be a reaction related to parasitic infection. However, further investigations would establish a definitive link between parasitic infections and ABO in this species. This case highlights the importance of studying rare medical conditions in wildlife, providing valuable insights into marine mammal health.

Phylogenetic and phylogeographic analyses of Anisakis simplex sensu stricto (Nematoda: Anisakidae) from the common minke whale in Korean waters.

The genus Anisakis is among the most significant parasites to public health, as it causes anisakiasis, a parasitic infection in humans resulting from consuming raw or undercooked seafood. Although the infection status of i>Anisakis in second intermediate hosts, such as marine fishes and cephalopods, and humans have been severally reported in Korea, no information about the definitive host in Korean waters is available. In 2014, 2 adult gastric nematodes were collected from a common minke whale (Balaenoptera acutorostrata) found in the East Sea, Korea. These worms were identified as A. simplex sensu stricto (s.s.) by comparing the mitochondrial COX2 marker with previously deposited sequences. Phylogenetic and phylogeographic analyses of A. simplex (s.s.) worldwide revealed 2 distinct populations: the Pacific population and the European waters population. This is the first report on adult i>Anisakis and its definitive host species in Korea. Further studies on Anisakis infection in other cetacean species and marine mammals in Korean seas are warrantedi>Anisakis.

Eight cases of canine thelaziosis found in two localities in Korea.

Thelazia callipaeda, a parasitic nematode that causes thelaziosis in various mammals, including humans, is known to be endemic in Korea. However, life cycle-related information on the parasite, primarily from human infection and a few dog cases, is limited. This study reports additional cases of T. callipaeda infections in dogs from both rural and urban areas in Korea, indicating the potential for transmission to humans and other animals. We collected 61 worms from 8 infected dogs from Paju and Cheongju Cities and observed their morphological characteristics under a light microscope. The findings indicate that T. callipaeda infections in animals in Korea may be underestimated and are distributed close to human environments. Our results contribute to the growing knowledge of the reservoir hosts of T. callipaeda in Korea and highlight the importance of continued surveillance and research to prevent and control this emerging zoonotic disease.

Molecular evidence of hookworms in public environment of Bangladesh.

Accurate diagnosis by precise identification of causative agents is essential for the effectiveness of any control interventions. Despite high zoonotic potential, available literature on hookworms in Bangladesh is still scarce and nonspecific. The objective of this study was to determine the occurrence of hookworms in public locations across northeastern Bangladesh (Sylhet metropolitan area) using integrated parasitological and molecular assays. A total of 130 samples (80 soil and 50 environmental canine feces) were collected and examined using modified flotation technique and formalin-ether sedimentation methods. Modified plate culture was used to isolate larvae. The identification was made based on morphometric features and confirmed by amplifying the ITS region of the nuclear rDNA. Overall, 66.2% (86/130) of examined samples were positive for hookworms infection. Characteristic eggs (61-68 × 29-37 µm) and/or larvae of hookworms were observed in 73.8% (59/80) soils and 54.0% (27/50) environmental fecal samples. Rhabditiform larvae (0.48-0.54 × 0.04-0.07 mm) were observed in cultured samples. Genetic analysis of rDNA sequences revealed the presence of Ancylostoma caninum and Ancylostoma ceylanicum. In this study, hookworms' contamination of the public environment was substantial. To the best of our knowledge, this is the first molecular proof of A. caninum and A. ceylanicum observed in urban public environment in Bangladesh.

First Report of Sarcocystis pilosa from a Red Fox (Vulpes vulpes) Released for the Re-Introduction Project in South Korea.

The red fox (Vulpes vulpes) is a known host for various parasites, including numerous helminths and protozoans. Among these, certain species in the genus Sarcocystis (phylum Apicomplexa) have been documented to possess the capability to infect red foxes as definitive hosts. In South Korea, red foxes have been introduced and released as part of a re-introduction program. However, two months after its release, one of the foxes was found dead because of illegal trapping. The fox was necropsied, and a subsequent coprological study revealed oocysts of Sarcocystis sp. in the intestinal contents. The oocysts were identified as Sarcocystis pilosa based on the 18S rRNA and cytochrome c oxidase subunit 1 (cox1) gene sequences. It exhibited a 99.7-100% identity with 18S rRNA gene sequences and 99.1-99.8% identity with cox1 gene sequences from other previously reported S. pilosa samples. Additionally, it showed identities of 95.4-96.4% and 91.1-91.5% with the cox1 gene sequences of S. hjorti and S. gjerdei, while demonstrating 99.6 and 98.1% identity with the 18S rRNA gene sequences of S. hjorti and S. gjerdei, respectively. This is the first report from mainland Asia, excluding the Japanese archipelago, indicating that the life cycle of S. pilosa persists in South Korea.

An update of intestinal helminth infections among urban slum communities in Bangladesh.

Aim: To assess the prevalence of intestinal helminth infections and associated risk factors among people living in urban slums in Bangladesh. Methods: A cross-sectional study was conducted across three clusters: Dhaka, Sylhet and Chattogram. In total, 360 individuals divided into two groups (school-aged children and adults) provided stool samples and completed a semi-structured questionnaire. Parasitological assessment was performed using the formol-ether concentration technique. Results: Overall, 31.7% (114/360) of participants had helminthiasis, with 13.3% (48/360) having mixed infections. Among the infected participants, school-aged children had a higher rate of infection (41.7%, 75/180) compared with adults (21.7%, 39/180). Ascaris lumbricoides was the predominant parasite, followed by hookworms, Trichuris trichiura, Hymenolepis nana, Enterobius vermicularis and Rhabditis sp. Parasitic infections were significantly associated with type of latrine used, direct exposure to soil, open defaecation, and presence of free-roaming animals. Conclusion: Despite continuous efforts to control helminthiasis, a substantial proportion of the study participants were infected with intestinal helminths. Ignorance of the roles of the environment and animals was influential, and had a negative impact on existing control interventions. An integrated public health and veterinary public health approach is required for sustainable control of intestinal helminthiasis.

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.

Morphological and Molecular Characterization of Philometroides seriolae from Japanese Amberjack Seriola quinqueradiata caught in East Sea, Republic of Korea.

The Japanese amberjack Seriolae quinqueradiata is one of the most consumed fish species among the Koreans. However, information regarding parasitic infection in Japanese amberjack is scarce. This study described the morphological and molecular characteristics of a species of philometrid nematode, Philometroides seriolae, which was recovered from Japanese amberjack. This fish was caught in the sea of Goseong-gun, Gangwon-do, Republic of Korea (Korea). Six P. seriolae (Nematoda: Philometridae) were recovered from 2 Japanese amberjacks. These parasites were subgravid female which were 325-420 mm long and 2.95-3.27 mm wide. Furthermore, they had typical papillae distributed on their body surface with 14 papillae at the apical view. Sequence analysis of the small subunits of ribosomal RNA (SSU rRNA) showed high sequence identity (99.8%, 1,607/1,611-bp) with that of P. seriolae (GenBank accession no. FJ155811). This nematode species has been newly added to the Korean nematode fauna.

Two Cases of Mange Mite (Sarcoptes scabiei) Infestation in Long-Tailed Goral (Naemorhedus caudatus) in Republic of Korea.

The long-tailed goral, Naemorhedus caudatus (Mammalia: Bovidae), is one of the endangered animals in the Republic of Korea (Korea). Sarcoptic mange mites infested in diverse species of mammals, including humans, but no case has been reported in long-tailed gorals. We report 2 cases of mange mite, Sarcoptes scabiei, infestation in long-tailed gorals. Mange mites were sampled in the skin legions of the 2 long-tailed gorals, which were rescued in 2 different regions, Uljin-gun, Gyeongsangbuk-do and Cheorwon-gun, Gangwon-do, Korea. Our results showed that the ectoparasite was the itch mite that burrowed into skin and caused scabies on the morphological inspection and placed within the phylogenetic relations of the species. The present study confirmed for the first time in Korea that mange mites are pathogenic scabies of long-tailed goral. Closer surveillance of this pathogenic ectoparasite in zoonotic and infectious ecosystems is warranted.

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.

First Clinical Cases of Spirometrosis in Two Cats in Korea.

This study reports the first two clinical cases of spirometrosis caused by Spirometra sp. in cats in Korea. In these two cases, the cats vomited, and long proglottids of tapeworm were recovered. The sick cats presented with anorexia and lethargy. However, they unexpectedly showed no diarrhea, which is the main symptom of spirometrosis. Based on a fecal floatation test as well as morphological and molecular analyses, the parasite was diagnosed as Spirometra sp. The 2 cases were treated with praziquantel. This study suggests regular monitoring of health and deworming in companion animals, even when animals are well cared for, with regular preventive medication. Additionally, spirometrosis should be considered in the differential diagnosis in cases of gastrointestinal symptoms in Spirometra endemic areas.

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.

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.

Integration of health education intervention to improve the compliance to mass drug administration for soil-transmitted helminths infection in Bangladesh: An implementation research.

INTRODUCTION: In Bangladesh, the prevention and control strategy of soil-transmitted helminthiasis (STHs) is based on the mass drug administration (MDA) program. Despite bi-annual MDA since 2008, the reported compliance is still below the target, and the STH prevalence is high in several areas. This study was done to assess the feasibility and barriers of integrating health education (HE) intervention to achieve the target MDA compliance in the local context of Bangladesh. MATERIALS AND METHODS: A mixed-method study, utilizing PRISM (Practical Robust Implementation Sustainability Model) framework, was conducted between July 2017 to March 2018 in Dhaka and Sylhet divisions of Bangladesh. A total of 640 school-aged children selected from four different schools were divided into intervention and control groups. Eight focus group discussions (FGDs) and eight in-depth interviews (IDIs) were also conducted among 56 adults, including parents of school-aged children, school teachers, and health officers. RESULTS: Quantitative findings revealed that HE intervention had a significant role (P < .05) to improve the mean knowledge score in the intervention group (3.35) compared to the control group (0.29). STH preventive behaviours and MDA participating attitudes were also significantly increased in the intervention group (P < .05) compared to the control group. Some of the major barriers associated with HE integration identified in the qualitative study were budget deficiencies, inadequate training of program implementers, and information gaps. In contrast, the school environment and positive community attitudes were observed as supportive factors for the integration of HE. CONCLUSION: Increased knowledge score and behaviour changes due to HE intervention demonstrated in this study hint that integration of HE with MDA is feasible and can be promising to promote MDA compliance and to reduce STH prevalence in this setting. However, the allocation of adequate budget, as well as coordination and collaboration with local political context, should be addressed for the sustainability of integration.

Descriptions of Two Digenean Trematodes Found from a Chinese Sea Snake, Laticauda semifasciata, in Republic of Korea.

We intended to describe 2 digenean trematodes found from a Chinese sea snake, Laticauda semifasciata, as the new fauna in the Republic of Korea. The snake was caught offshore of Aewol-eup, Jeju-do, in August 2017. Two species of fluke were found in the lung and intestinal tract of the snake in the process of necropsy. They were identified as Pulmovermis cyanovitellosus Coil and Kuntz, 1960 and Harmotrema laticaudae Yamaguti, 1933, respectively based on the morphological characters. Pulmovermis cyanovitellosus showed elongated body with well-developed and elongated male genital system and compact vitelline. And H. laticaudae was characterized by linguiform body with heavily armed cirrus with excretory system. This is the first time both species have been reported and described off the Korean coast. We provide morphological descriptions with some comments on their biology and geographical distribution. In addition, the taxonomic validity of the genus Hydrophitrema Sandars, 1960 was discussed, in terms of morphological descriptions and host ranges. This study provides novel insight into digenean fluke species existing off the coast of Korea.

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.