Zoonotic Genotypes of Enterocytozoon bieneusi in Wild Living Invasive and Native Carnivores in Poland

Wild carnivores, both introduced and native species, are able to adapt well to peri-urban environments, facilitating cross-species pathogen transmission with domestic animals, and potentially humans. The role of wild living reservoir hosts cannot be ignored because of their known carriage of E. bieneusi zoonotic genotypes. In the past decades, populations of wild living carnivores, i.e., native, such as red foxes, and invasive, such as raccoon dogs and raccoons, have increased and adapted to synanthropic environments across Europe, including Poland. The knowledge concerning E. bieneusi genotype identification and distribution in wild carnivores is limited worldwide. A total of 322 individual fecal samples from six carnivore species, i.e., raccoon, raccoon dog, red fox, European badger, pine and beech martens, were collected and then analysed for the presence of E. bieneusi using the nested PCR method. Overall prevalence of the pathogen was estimated to be as high as 27.3%. The infection rates for E. bieneusi varied between the carnivore species, from 13.7% in beech martens to 40.4% in raccoon dogs. Based on sequence analysis of the ITS region of the rRNA gene marker, we detected five known genotypes of E. bieneusi in examined animals. In the invasive species, E. bieneusi NCF2 and D genotypes have been identified, whereas in the native ones, E. bieneusi NCF2, D, C, EbCar2 and Type IV genotypes were identified. All E. bieneusi genotypes recorded in this survey clustered in Group 1, showing their zoonotic potential. Our results provide the first description of the occurrence and genotypes of the microsporidian E. bieneusi in wild living population of raccoon dogs in Europe. Our findings are important for the study of pathogen epidemiology and emphasize the fact that the invasive and the native wild living carnivores, both widely distributed, should be considered more seriously as significant sources of zoonotic pathogens hazardous to domestic and farmed animals and humans.

Factors and features of manifestation and distribution of rabies on the territory of the Republic of Tuva

The dynamics and features of animal rabies manifestation on the territory of the Republic of Tuva have been studied. In the Republic of Tuva in the period of 1979-2019 outbreaks of rabies were reported in 1979, 1987, 2007-2009, 2012-2014, 2018-2019. There were 79 adverse locations revealed with 127 laboratory confirmed cases of rabies of four species of wild animals, two domestic carnivores and four agricultural animals. For the first time the cause of the outbreak of rabies was the introduction of infection by wild carnivores from adverse neighboring regions and Mongolia. Subsequently, a natural focus of rabies developed on the territory of the region. The epidemic process of rabies proceeded with five waves of intensity exacerbation, which was facilitated by a sharp increase in the population of wild carnivores, especially wolves, as well as an uncontrolled population of stray dogs with the involvement of domestic and farm animals in the epizootic process. The spread of the disease is influenced by the natural and climatic conditions of the region, the traditional distant pasture management of livestock with the movement of livestock to pastures adjacent to the habitat of wild predators. Domestic and farm animals are attacked by predators, most often wolves and foxes. Infection of animals and humans with rabies occurs through bites with saliva. As a result, rabies foci appear in these areas. Zones of high and low degree of epizootic danger and zones free from rabies have been identified on the territory of the Republic of Tuva. The high-risk areas include the steppe and semi-desert territories of the Ubsu-Nur and Tuva depressions. The mountain-taiga Todzhinsky and Tere-Kholsky regions of the eastern zone are classified as areas of low epizootic danger; Pii-Khem and Kaa-Khem regions of the Yenisei basin of the central zone are classified as rabies-free.

Rabies eradication measures in wildlife in the Republic of Moldova

Introduction. Rabies is one of the most dangerous zoonoses in the world with a lethality rate around 100% of infected animals. The main factor in rabies’ control is the prophylactic immunization of domestic and wild carnivores. Material and methods. As a research material for this manuscript, a number of rabies cases in wildlife over the last 5 years was studied, as well as some data from the national program regarding the vaccination of wildlife animals in the Republic of Moldova. Results. The anual incidence of rabies in wildlife was determined to range between 12 to 25% of the total number of rabies cases at national level. Due to vaccination by combining some methods (aerial and manual), the incidence of positive cases of rabies in wildlife has decreased by 48% in the last 5 years. Conclusions. The use of the mixed vaccination method of wildlife animals (manual and the distribution of baits by plane) contributed significantly to reducing the incidence of rabies in wildlife.

Molecular Characteristics of Carnivore protoparvovirus 1 with High Sequence Similarity between Wild and Domestic Carnivores in Taiwan

Carnivore protoparvovirus 1 (CPPV-1) is a DNA virus causing gastrointestinal disease and immunosuppression in various terrestrial carnivores. Domestic dogs and cats are considered the primary CPPV-1 reservoirs. The habitat overlap of wild carnivores and free-roaming dogs increases the threat of CPPV-1 transmission between them. This study explored the CPPV-1 distribution among wild carnivores in Taiwan through PCR screening and compared the partial capsid protein (VP2) gene sequences from wild and domestic carnivores. In total, 181 samples were collected from 32 masked palm civets (Paguma larvata), 63 Chinese ferret badgers (Melogale moschata), and 86 crab-eating mongooses (Herpestes urva), from 2015 to 2019 were screened for CPPV-1. The average prevalence of CPPV-1 was 17.7% (32/181), with the highest prevalence in masked palm civets (37.5%). In addition, a masked palm civet was coinfected with two CPPV-1 strains. Among the 33 partial VP2 gene sequences, 23 were identical to the sequences amplified from domestic dogs and cats in Asia, and the remaining 10 were identified for the first time. This study supported the circulation of CPPV-1 strains with the same genomic features as domestic carnivores that are also in wild carnivores from the same environment in Taiwan by molecular data. Therefore, further population control and health management of free-roaming domestic carnivores are recommended.

Molecular survey of Besnoitia spp. (Apicomplexa) in faeces from European wild mesocarnivores in Spain. Short running title: Molecular survey of Besnoitia spp in faeces from wild carnivores.

Numerous studies have unsuccessfully tried to unravel the definitive host of the coccidian parasite Besnoitia besnoiti. Cattle infections by B. besnoiti cause a chronic and debilitating condition called bovine besnoitiosis that has emerged in Europe during the last two decades, mainly due to limitations in its control associated to the absence of vaccines and therapeutical tools. Although the exact transmission pathway of B. besnoiti is currently unknown, it is assumed that the parasite might have an indirect life cycle with a carnivore as definitive host. Current lack of studies in wildlife might underestimate the importance of free-living species in the epidemiology of B. besnoiti. Thus, the aim of the present study is to assess the presence of Besnoitia spp. in free-ranging mesocarnivores in Spain. DNA was searched by PCR on faeces collected from wild carnivores as a first approach to determine which species could be considered as potential definitive host candidates in further research. For this purpose, a total of 352 faecal samples from 12 free-living wild carnivore species belonging to the Canidae, Felidae, Herpestidae, Mustelidae, Procyonidae, and Viverridae families were collected in seven Spanish regions. PCR testing showed that Besnoitia spp. DNA was present in four faecal samples from red foxes collected in western Spain, an area with the greatest density of extensively reared cattle and associated to high incidence of bovine besnoitiosis in the country. To date, this is the first report of a Besnoitia besnoiti-like sequence (99.57% homology) from carnivore faeces in a worldwide context. Red foxes might contribute to the epidemiology of B. besnoiti, although further studies, mostly based on bioassay, would be needed to elucidate the accuracy and extent of these interesting findings.