Molecular Detection and Identification of Piroplasm in Cattle from Kathmandu Valley, Nepal.

BACKGROUND: Tick-borne protozoan parasites (TBPPs) cause significant problems for domestic animals' health in Nepal. TBPPs are routinely diagnosed by labor-intensive blood smear microscopy. In Nepal, there are some reports of Babesia and Theileria in cattle, although species identification is rarely performed. Therefore, we performed conventional nested PCR (nPCR) followed by sequence analysis to identify TBPP species infecting cattle in Nepal. METHODS: One hundred and six blood samples were collected from cattle in the Kathmandu Valley. Thin blood smears were prepared for microscopic examination. Parasite DNA was extracted from the blood, and nPCR and sequencing were performed to identify the TBPPs present. RESULTS: Among the 106 samples, 45 (42.5%) were positive for piroplasm (Babesia spp. and Theileria spp.) via microscope observation and 56 (52.8%) samples were positive via nPCR. The obtained PCR products were used for direct sequencing, and we identified the species as B. bigemina, B. bovis, T. annulate and T. orientalis. Phylogenetic analyses showed that the B. bovis, B. bigemina and T. orientalis sequences from this study belonged to each species clade. On the other hand, T. annulate was divided into two clades in the analysis, and our T. annulate sequences were also divided in these two clades. The piroplasm-positive cattle showed lower hemoglobin and red blood cells than healthy cattle. CONCLUSIONS: To the best of our knowledge, this study is the first to apply molecular detection and species determination of TBPPs in cattle in Nepal. The results of this study may be used as a starting point for the development of successful TBPP surveillance and prevention programs in Nepal.

Gastrointestinal parasites of zoonotic importance detected in porcine faeces in Chitwan National Park, Nepal.

Wild boar (Sus scrofa) is considered to be a potential source of zoonotic parasites. Wild boars are found at considerable number in and around the Chitwan National Park (CNP). The information regarding their intestinal parasites is limited. A cross-sectional study was carried out to determine the prevalence of gastrointestinal parasites in wild boars in CNP. A total of 100 fresh fecal samples were subjected to microscopic examination using direct smear, floatation and sedimentation method. Overall, 95% fecal samples were found positive for at least one parasite. Prevalence of protozoan parasites was found comparatively higher (70%) followed by nematode (56%) and trematode (12%). Nine gastrointestinal parasites such as Eimeria sp. (70% without micropyle and 40% with micopyle), Fasciola sp. (12%), Strongyloides sp. (56%), strongyle-type nematodes (49%), Stephanurus sp. (44%), Globocephalus sp. (38%), Metastrongylus sp. (12%), Ascaris sp. (7%) and Trichuris sp. (6%). were recorded. Eimeria sp. exhibited the highest prevalence while Trichuris had the least prevalence. This study has provided baseline information regarding the diversity of gastrointestinal parasites in wild boars. It requires continuous study at molecular level to explore other species of parasites and verify their zoonotic potential.

Minimum risk of intestinal parasitic infection to the visitors of Ghandruk, Kaski, Nepal: a pilot survey.

Ghandruk is one of the famous destinations of both international and domestic tourists situated in Kaski, Nepal. Travel-related diseases are an important aspect that one should consider before making a travel plan. Among diseases, zoonotically important ones make serious worries among visitors. In order to assess the existence of the zoonotically important parasitic disease in Ghandruk, a pilot survey was carried out by examining representative stool samples (n=51) of local residence, domestic animals, pet animals in Ghandruk. Samples were examined using direct smear as well as concentration methods. A questionnaire survey was conducted to see the associated risk factors among residents and their livestock of Ghandruk. None of the faecal samples from residents (n=14) found positive for any kind of intestinal parasites (IPs), while samples from most of the livestock: chicken (86%, 6/7), pigeons (75%, 3/4), cow (66%, 2/3), mule (60%, 3/5), and dog (60%, 3/5) showed heavy infection, except goat and buffalo indicated no infection. Eimeria spp., Ascardia spp. and cestodes spp. were the most prevalent IPs in livestock. Periodic deworming, walking outdoor with sandals/shoes, frequent use of soap and water for handwashing as reported by most of the residents (>80%) could be the main reason behind zero prevalence of IPs in them. The heavy infection among livestock may be incriminated to the contaminated vicinity and free-range livestock and poultry which were noticed in contact with river, sludge, and toilets during our field observation. Conclusively, it indicates that the residents of Ghandruk are conscious about their health, but have not paid satisfactory attention to the hygiene of their domestic animals including livestock, poultry and even pet. Though the observed parasites in livestock are of minimal zoonotic importance regarding safety of visitors, it is deemed necessary for at least to apply some preventive measures to mitigate the burden of parasites in their animals.

Prevalence of gastrointestinal parasites among greater one-horned rhino in Chitwan National Park, Nepal.

Recently, many individuals of greater one-horned rhino (GOHR) were died with unknown reason in Chitwan National Park (CNP), Nepal. This has arisen chaos and confusion in the rhino conservation program of the country. This study was designed to determine the prevalence of gastrointestinal parasites among GOHR in the CNP. A total of 100 dung samples were collected opportunistically by the random sampling method. Dung samples were preserved in 2.5% potassium dichromate solution and analysed in the laboratory by the direct smear and concentration method (floatation and sedimentation). Eggs and larvae of gastrointestinal parasite were found in 91% examined samples. Altogether 13 different genera of parasites were identified with one protozoan i.e. Eimeria sp. (9%), nine nematodes i.e. Strongyloides sp. (65%), Ascaris sp. (16%), Haemonchus sp. (15%), Dromeostrongylus sp. (9%), Oxyuris sp. (8%), Bunostomum sp. (8%), Chabertia sp. (5%), Trichostrongylus sp. (4%) and Nematodirus sp. (2%), one cestode i.e. Anoplocephala sp. (16%) and two trematodes i.e. Paramphistomum sp. (31%) and Fasciola sp. (14%). Nematode parasites were found to be most prevalent (87% of samples) followed by trematodes (45%), cestodes (16%) and protozoans (9%). The study indicates a high prevalence of gastrointestinal parasites in the GOHR of CNP and identifies that there is need of strategic control measures to protect this endangered species from parasitic infection.

Livestock Depredation by Leopards and Tigers Near Bardia National Park, Nepal.

Wildlife attacks on livestock near human settlements are increasing due to the proximity of humans to protected areas. These attacks are often severe due to depredations of livestock adversely affecting the livelihoods of people. The nature of carnivore depredations on livestock can differ based on the carnivore species, animal husbandry practices, season, and deterrent technique. We surveyed people living near Bardia National Park (BNP), Nepal, to compare hoofed livestock depredations by leopards (Panthera pardus) and tigers (P. tigris) near (<1 km) and far (>1 km) from this protected area. Overall, 1476 hoofed livestock were reportedly depredated by leopards, and 209 by tigers, during 2015-2019. The number of hoofed livestock killed by leopards each season was, at least, 86% higher than the number killed by tigers. More livestock were killed at BNP irrespective of carnivore deterrent techniques used. Due to severe effects created by livestock depredations near BNP, we recommend using more efficacious deterrent techniques when practical, in addition to improved livestock husbandry practices such as night penning.

Prevalence of intestinal parasites in Deula community, Kirtipur Municipality, Kathmandu, Nepal.

Infections with intestinal protozoans, helminths, and other parasites are major, but overlocked problems in developing countries. Accurate surveys of intestinal parasites in Deula community inform empirical treatment regimens and can assess the impact of community-based drug treatment programs. There is limited information on this topic in Nepal. In a prospective study of intestinal parasites in symptomatic people of the Deula community, January-July 2018, samples were examined by microscopy of a direct and concentrated faecal sample. We studied 150 samples. The median (inter-quartile range) age of the people was 34.5 (16-50.5) years, 52% were female and 48% were male. The proportion of people with abdominal pain was 30.39%, diarrhea 42.16%, anemia 14.71% and malnutrition 3.92%. Altogether 102 (68%) parasitic infections were detected. The most common parasites using all methods of detection were Entamoeba histolytica (35.30%), Giardia lamblia (21.57%), Trichuris trichiura (14.71%), hookworm (13.73%), Ascaris lumbricoides (7.84%) and Strongyloides stercoralis (6.85%). E. histolytica was most common, followed by G. lamblia in all age groups, T. trichiura and A. lumbricoides. Hookworm and S. stercoralis were more common with increasing age. This study substantiate the significance of intestinal parasitic infections in indicative Deula community and the need for adequate facilities for laboratory diagnosis together with education to improve personal hygiene and sanitation.