Identification of Novel Feline Paramyxoviruses in Guignas (Leopardus guigna) from Chile

The family of paramyxoviruses has received growing attention as several new species have been identified recently, notably two different clusters in domestic cats, designated as feline morbillivirus (FeMV) and feline paramyxovirus (FPaV). Their phylogenetic origin and whether wild felids also harbor these viruses are currently unknown. Kidney samples from 35 guignas (Leopardus guigna), a wild felid from Chile, were investigated for paramyxoviruses using consensus-RT-PCR. In addition, thirteen serum samples of guignas were screened for the presence of FeMV-specific antibodies by an immunofluorescence assay (IFA). Viral RNA was detected in 31% of the kidney samples. Phylogenetic analyses revealed two well-supported clusters, related to isolates from domestic cats, rodents and bats. No significant histopathology changes were recorded in infected guignas. Serology identified two samples which were positive for FeMV-specific antibodies. Our study highlights the diversity of paramyxovirus infections in felids with special emphasis on guignas from Chile

Large-scale survey for canine vector-borne parasites in free-ranging dogs and foxes from six diverse bioclimatic regions of Chile

Chile is a large country with a marked range of climate conditions that make it an ideal scenario for the study of vector-borne parasites (VBPs); however, knowledge about their distribution is limited to a few confined areas of this country. The presence of Hepatozoon spp., piroplasmids, Leishmania spp. and filarioids was investigated through molecular and serological methods in blood and serum samples of 764 free-ranging rural dogs, 154 Andean foxes (Lycalopex culpaeus), and 91 South American grey foxes (Lycalopex griseus) from six bioclimatic regions across Chile. Hepatozoon spp. DNA was exclusively detected in foxes (43% prevalence), including sequences closely related to Hepatozoon felis (24.1%; only Andean foxes), Hepatozoon americanum (16.2%; only grey foxes), and Hepatozoon canis (1.25%; in one grey fox). Risk factor assessment identified a higher probability of Hepatozoon infection in juvenile foxes. DNA of piroplasmids was detected in 0.7% of dogs (Babesia vogeli) but in no fox, whilst antibodies against Babesia sp. were detected in 24% of the dogs and 25% of the foxes, suggesting a wider circulation of canine piroplasmids than previously believed. A positive association between the presence of antibodies against Babesia and high Rhipicephalus sanguineus sensu lato burden was observed in dogs. Leishmania spp. DNA and antibodies were detected in 0.8% and 4.4% of the dogs, respectively. Acanthocheilonema reconditum was the only blood nematode detected (1.5% of the dogs and no fox). Differences in prevalence among bioregions were observed for some of the VBPs. These results expand our knowledge about the occurrence of vector-borne parasites in Chile, some of which are firstly reported herein. This information will facilitate the diagnosis of vector-borne diseases in domestic dogs and improve the control measures for both domestic and wild canids

Distribution, transmission, and risk factors of infection with canine vector-borne pathogens in wild foxes and free-ranging dogs in Chile

Canine vector-borne pathogens (CVBP) distribution tends to be circumscribed to certain areas with climatic features where their vectors are able to persist. In this aspect, Chile possesses a marked variety of bioclimates that provides an ideal scenario to study the distribution of vector-borne pathogens, considering that ticks of the Rhipicephalus sanguineus species group are widely distributed in the country. Despite this, information about the presence, distribution, and impact of CVBP in domestic and wild canines in Chile is very scarce. In this study, blood and serum samples of 981 dogs, 155 Andean foxes (Lycalopex culpaeus), and 90 grey foxes (L. griseus) were obtained from six different bioclimatic regions throughout Chile.Para acceder a la videoconferencia completa, hacer clic en "Enlace externo".Sociedad Latinoamericana de Ecología de Vectore

Wild deer (Pudu puda) from Chile harbor a novel ecotype of Anaplasma phagocytophilum

[Background] Deer species play an important role in the enzootic cycles of several Anaplasma species. While in the Northern Hemisphere ticks of genus Ixodes are well recognized vectors of these intracellular bacteria, less is known regarding the biological cycles of Anaplasma spp. in South America.[Methods] Using PCR protocols and Sanger sequencing, we assessed the presence of Anaplasma spp. in blood and ticks collected on a native deer species (Pudu puda) from southern Chile.[Results] Based on phylogenetic analyses of the 16S rRNA, gltA and groEL genes and calculation of average sequence divergence for groEL, our results bring to light a novel genovariant of Anaplasma phagocytophilum (named strain “Patagonia”). The strain represents a novel ecotype within the A. phagocytophilum species complex and was detected in both P. puda and their ticks. Using a larger matrix, denser taxon sampling and outgroup, our maximum-likelihood- and Bayesian-inferred phylogenies for groEL provide an accurate picture of the topology of A. phagocytophilum ecotypes and their evolutionary relationships.[Conclusions] This is the first report of an ecotype of A. phagocytophilum in South America. Our results provide novel insight into the genetic diversity and ecology of this complex of bacterial lineages. Further studies should elucidate the enzootic cycle of A. phagocytophilum strain “Patagonia” and assess its pathogenic potential for pudues, domestic animals and humans in the region.This study was funded by the “Fondo Nacional de Desarrollo Científico y Tecnológico (FONDECYT)” No. 11220177, and by the ANID BECAS/Scholarship Program/DOCTORADO NACIONAL/2019–21190078 and 2020–21200182. Funders had no role in the study design, data collection, analysis, preparation of the manuscript and decision to publish. The contribution of JEU was funded by the Atracción Talento de la Comunidad de Madrid Fellowship Program (REFF 2019-T2/AMB-13166).Peer reviewe

Epidemiology and molecular characterization of Carnivore protoparvovirus-1 infection in the wild felid Leopardus guigna in Chile

14 Pág. Centro de Investigación en Sanidad Animal (CISA)Landscape anthropization has been identified as one of the main drivers of pathogen emergence worldwide, facilitating pathogen spillover between domestic species and wildlife. The present study investigated Carnivore protoparvovirus-1 infection using molecular methods in 98 free-ranging wild guignas (Leopardus guigna) and 262 co-occurring owned, free-roaming rural domestic cats. We also assessed landscape anthropization variables as potential drivers of infection. Protoparvovirus DNA was detected in guignas across their entire distribution range, with observed prevalence of 13.3% (real-time PCR) and 9% (conventional PCR) in guignas, and 6.1% (conventional PCR) in cats. Prevalence in guigna did not vary depending on age, sex, study area or landscape variables. Prevalence was higher in juvenile cats (16.7%) than in adults (4.4%). Molecular characterization of the virus by amplification and sequencing of almost the entire vp2 gene (1,746 bp) from one guigna and five domestic cats was achieved, showing genetic similarities to canine parvovirus 2c (CPV-2c) (one guigna and one cat), feline panleukopenia virus (FPV) (one cat), CPV-2 (no subtype identified) (two cats), CPV-2a (one cat). The CVP-2c-like sequence found in a guigna clustered together with domestic cat and dog CPV-2c sequences from South America, suggesting possible spillover from a domestic to a wild species as the origin of infection in guigna. No clinical signs of disease were found in PCR-positive animals except for a CPV-2c-infected guigna, which had haemorrhagic diarrhoea and died a few days after arrival at a wildlife rescue centre. Our findings reveal widespread presence of Carnivore protoparvovirus-1 across the guigna distribution in Chile and suggest that virus transmission potentially occurs from domestic to wild carnivores, causing severe disease and death in susceptible wild guignas.Comisión Nacional de Investigación Científica y Tecnológica, Grant/Award Number: Fondecyt Iniciación 11150934 and PAI 77190064; National Geographic Society, Grant/Award Number: C309-15; Morris Animal Foundation, Grant/Award Number: D15ZO-413Peer reviewe

Molecular detection and characterization of hemoplasmas in the pudu (Pudu Puda), a native CERVID from CHILE

8 Pág. Centro de Investigación en Sanidad Animal (CISA)Hemotropic mycoplasmas cause hemolytic anemia in a variety of wild and domestic mammals. Despite growing evidence about their widespread presence and genetic diversity in wildlife, their presence has never been investigated in Chilean artiodactyls. We aimed to describe the presence and diversity of hemoplasmas in pudus (Pudu puda), a small cervid native to Chile. Hemoplasma infection was assessed in blood samples from 43 wild and 33 captive pudus from central and southern Chile by direct sequencing of the 16S rRNA gene. We detected hemoplasmas in 13%, with no statistical differences between wild (19%) and captive animals (6%). A sequence closely related to Mycoplasma ovis was present both in wild (14%) and captive (6%) pudus. Two previously undescribed sequences, classified in a clade including hemoplasmas from carnivores, were found in one wild pudu each. This study presents the first evidence of the presence of M. ovislike organisms in Chile and of the susceptibility of pudus to infection with hemoplasmas. Further research is needed to understand the pathologic consequences of this pathogen for pudus, its effects at the population level, and their potential impact on the health small ruminants and other wildlife species in Chile.Peer reviewe

Molecular survey and phylogenetic analysis of Bartonella sp., Coxiella sp., and hemoplamas in pudu (Pudu puda) from Chile: first report of Bartonella henselae in a wild ungulate species

IntroductionRecent evidence shows a high diversity of infectious agents in wildlife that represent a threat to human, domestic, and wild animal health. In Chile, wild populations of the most common cervid species, pudu (Pudu puda), have been reported as hosts for novel pathogens such as Mycoplasma ovis-like and a novel ecotype of Anaplasma phagocytophilum. A better understanding of the epidemiology of this group and other intracellular bacteria that might have cervids as hosts would enlighten their population relevance. This study aimed to determine the occurrence and genetic diversity of Bartonella spp., hemotropic mycoplasmas, and Coxiella burnetii in pudus from Chile.MethodsThe DNA was extracted from the blood samples of 69 wild free-ranging and 30 captive pudus from Chile. A combination of real-time (nouG gene for Bartonella and IS1111 element for C. burnetii) and conventional PCR (16S rRNA for hemotropic Mycoplasma spp. and rpoB, gltA, and ITS for Bartonella spp.) was used for pathogen screening and molecular characterization.ResultsDNA of Bartonella spp. was detected in 10.1% [95% CI (5.2–18.2%)] samples, hemotropic Mycoplasma spp. in 1.7% [95% CI (0.08–10.1%)], and C. burnetii in 1.0% [95% CI (0.05–6.3%)] samples. Two sequenced samples were identified as Mycoplasma ovis-like, and one free-ranging pudu was positive for C. burnetii. While one captive and two free-ranging pudus were positive for Bartonella henselae, one wild pudu was co-positive for B. henselae and Bartonella sp., similar to Bartonellae identified in ruminants.DiscussionTo the best of our knowledge, this is the first report of B. henselae in wild ungulate species, and C. burnetii and Bartonella spp. in wild ungulate species in South America. Further research will be necessary to evaluate the potential role of pudu as reservoirs of infection and identify the sources for disease transmission among humans and wild and domestic animals

Lack of Exposure to Mycobacterium bovis and Mycobacterium avium subsp. paratuberculosis in Chilean Cervids, and Evidence of a New Mycobacterium-Like Sequence

5 Pág. Centro de Investigación en Sanidad Animal (CISA)Screening of serum and fecal samples from huemul (Hippocamelus bisulcus) and pudu (Pudu puda) from southern Chile for Mycobacterium bovis and Mycobacterium avium paratuberculosis (MAP) found all but four samples Mycobacterium-negative. The positive sequences showed only 92-93% similarity with MAP and were from remote Isla Riesco populations.This work was supported by Asociacion Kauyeken through Fondo Nacional de Desarrollo Regional Project 5961 and the J. M. Kaplan Fund, Buin Zoo, NGO Chiloe Silvestre, Universidad San Sebastian-Sede Puerto Mont and ANID/CONICYT Fondecyt 3160056 and 11181180 (to D.M.-A.), and Estancia Anita BeatrizPeer reviewe

Mycobacterium tuberculosis sensu stricto in African Apes, What Is Its True Health Impact?

Since the Symposium on Mycobacterial Infections of Zoo Animals held at the National Zoological Park, Smithsonian Institution in 1976, our understanding of tuberculosis (TB) in non-domestic animals has greatly expanded. Throughout the past decades, this knowledge has resulted in improved zoo-habitats and facilities design, stricter biosecurity measures, and advanced diagnostic methods, including molecular techniques, that have significantly decreased the number of clinical disease caused by Mycobacterium tuberculosis in apes under human care settings. In the other hand, exponential growth of human populations has led to human encroachment in wildlife habitat which has resulted in increased inter-species contact and recurrent conflict between humans and wild animals. Although it is widely accepted that non-human primates are susceptible to M. tb infection, opinions differ with regard to the susceptibility to develop disease amongst different taxa. Specifically, some authors suggest that African apes are less susceptible to clinical tuberculosis than other species of primates. The aim of this review article is to evaluate the current scientific literature to determine the actual health impact of disease caused by Mycobacterium tuberculosis and more specifically Mycobacterium tuberculosis sensu stricto in African apes. The literature review included literature databases: Web of Science, Pubmed, Scopus, Wiley, Springer and Science direct, without temporal limit and proceedings of annual conferences in the field of wildlife health. Our general inclusion criteria included information about serological, molecular, pathological (macroscopic and/or microscopic), and clinical evidence of TB in African apes; while our, our more stringent inclusion selection criteria required that in addition to a gross pathology, a molecular test confirmed Mycobacterium tuberculosis sensu stricto as the cause of disease or death. We identified eleven reports of tuberculosis in African apes; of those, only four reports met the more stringent selection criteria that confirmed M. tb sensu stricto in six individuals. All reports that confirmed M. tb sensu stricto originated from zoological collections. Our review suggests that there is little evidence of disease or mortality caused by M. tb in the different species of African apes both under human care and free ranging populations. Additional studies are needed in free-ranging, semi-captive populations (sanctuaries) and animals under human care (zoos and rescue centers) to definitely conclude that this mycobacteria has a limited health effect in African ape species

Detection of persistent pestivirus infection in pudú (Pudu puda) in a captive population of artiodactyls in Chile

Background: Bovine Viral Diarrhea Virus (BVDV) is the viral agent causing the most important economic losses in livestock throughout the world. Infection of fetuses before their immunological maturity causes the birth of animals persistently infected with BVDV (PI), which are the main source of infection and maintenance of this pathogen in a herd. There is evidence of susceptibility to infection with BVDV in more than 50 species of the order Artiodactyla, and the ability to establish persistent infection in wild cervid species of South America could represent an important risk in control and eradication programs of BVDV in cattle, and a threat to conservation of these wild species. In this study, a serological and virological study was performed to detect BVDV infection in a captive population of non-bovine artiodactyl species in a Chilean zoo with antecedents of abortions whose pathology suggests an infectious etiology. Results: Detection of neutralizing antibodies against BVDV was performed in 112 artiodactyl animals from a zoo in Chile. Three alpacas (Vicugna pacos), one guanaco (Lama guanicoe) and seven pudus (Pudu puda) resulted seropositive, and the only seronegative pudu was suspected to be persistently infected with BVDV. Then two blood samples nine months apart were analyzed by a viral neutralization test and RT-PCR. Non-cytopathogenic BVDVs were isolated in both samples. A phylogenetic analysis showed that the virus was highly related to BVDV-1b strains circulating among Chilean cattle. Conclusions: This is the first report of a South American deer persistently infected with Bovine Viral Diarrhea Virus. Further studies are needed to determine the possible role of BVDV as a pathogen in pudus and as a threat to their conservation.Laboratory of Animal Virology, Faculty of Livestock and Veterinary Sciences, University of Chile Department of Conservation and Research, Buin Zoo Zoological Park, Chil