An Integrated Coproscopic and Molecular Method Provides Insights into the Epidemiology of Zoonotic Intestinal Helminths of Dogs across Cambodia

Abstract

Introduction. In Cambodia, a limited number of focal surveys targeting dogs in rural communities have reported intestinal helminths of potential zoonotic risk as highly endemic. In this study, we investigated the prevalence, diversity, distribution, and risk predictors associated with zoonotic intestinal helminths infecting pet, community, and pagoda dogs across rural and urban settings in Cambodia through an integrated coproscopic and molecular approach. Methods. Faecal samples, demographic, and clinical data were collected from 457 dogs across Cambodia. Faeces were screened for gastroenteric parasites using sodium nitrate faecal floatation (1.30 SpGr) and multiplex TaqMan-based quantitative polymerase chain reaction (qPCR) assays for differentiation of canine hookworm species (Ancylostoma ceylanicum, Ancylostoma braziliense, Ancylostoma caninum, and Uncinaria stenocephala) and for Strongyloides spp. Conventional polymerase chain reaction (PCR) and DNA sequencing was used to further characterise eggs resembling zoonotic helminths that could not be designated to a species level by light microscopy alone. In addition, infection intensities for hookworms and Toxocara as eggs per gram of faeces were estimated to assess the dog age-dependent contribution of these zoonotic parasites in the environment. Finally, multiple logistic regression analyses were performed to identify risk predictors for gastrointestinal parasitoses. Results. Using combined coproscopic and molecular methods, we detected at least one helminth in 79% of the sampled dogs. Of these, 78.8% were infected with at least one zoonotic species, with A. ceylanicum (50%) and A. caninum (50%) constituting the most common parasites followed by A. braziliense (20%), Toxocara canis (15%), Strongyloides spp. (5%), Dipylidium caninum (2%), and Eucoleus spp. (2%). The indirect zoonotic helminths Spirometra erinaceieuropaei, Spirometra mansoni, and Haplorchis yokogawai contributed to 14% of the total infections in dogs. Dogs were also found mechanically passing eggs of large trematodes such as Fischoederius elongatus, Schistosoma spp., Paramphistomatidae, and Gastrothylax crumenifer. Contradictory to previous reports, the prevalence of T. canis was highest in adult dogs with egg shedding intensity peaking in dogs more than 7 years of age. Finally, we found that pale mucous membranes and low total protein were predictors of hookworm infection in dogs ( $p<0.05$ ). Conclusion. Dogs within both rural and urban settings across Cambodia are highly infected with a large diversity of zoonotic gastroenteric helminths. This study reports the presence of A. braziliense, the aetiological agent of hookworm-related “creeping eruptions” for the first time in Cambodia. Moreover, infection intensity data suggest that adult dogs should also be targeted by deworming campaigns to reduce the prevalence of zoonotic hookworm and T. canis infections within the country. These results highlight the need for an integrated approach to canine population management and parasite control in dogs across urban and rural settings in Cambodia to mitigate the public health risks and impacts posed by these helminths.