Molecular survey of selected viruses in Pudus (Pudu puda) in Chile revealing first identification of caprine herpesvirus-2 (CpHV-2) in South American ungulates.

Viral diseases jeopardize the health of wildlife in Chile. However, this country lacks health surveillance programs that allow for defining preventive measures to tackle such diseases. The objective of this study was to determine the occurrence and the genetic diversity of pestivirus, herpesvirus and adenovirus in pudus from Chile. Blood samples from wild (n=34) and captive (n=32) pudus were collected between 2011 and 2019 and analyzed through consensus PCR. All the samples were negative to pestivirus and adenovirus. Herpesvirus was confirmed in four captive, and one wild pudu. All four zoo animals share the same sequence for both polymerase and glycoprotein genes. Both sequences share a 100% identity with caprine herpesvirus-2, classifying them in the same cluster as the Macavirus group. In turn, novel sequences of the polymerase and glycoprotein B genes were obtained from the wild pudu. Our study reports the first evidence of CpHV-2 infection in Chile and South American ungulate populations. Further research will be necessary to assess the pathogenicity of CpHV-2 in this species. It is also urgently recommended that molecular, serological and pathological screening should be conducted in Chilean wild and captive pudus to understand the impact of the herpesvirus on their populations.

MOLECULAR DETECTION AND CHARACTERIZATION OF HEMOPLASMAS IN THE PUDU (PUDU PUDA), A NATIVE CERVID FROM CHILE.

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.

SEROSURVEY FOR SELECTED PARASITIC AND BACTERIAL PATHOGENS IN DARWIN'S FOX (LYCALOPEX FULVIPES): NOT ONLY DOG DISEASES ARE A THREAT.

The Darwin's fox (Lycalopex fulvipes) is one of the most endangered carnivores worldwide, with the risk of disease spillover from domestic dogs being a major conservation threat. However, lack of epidemiologic information about generalist, non-dog-transmission-dependent protozoal and bacterial pathogens may be a barrier for disease prevention and management. To determine the exposure of some of these agents in Darwin's fox populations, 54 serum samples were collected from 47 Darwin's foxes in Southern Chile during 2013-18 and assessed for the presence of antibodies against Brucella abortus, Brucella canis, Coxiella burnetii, pathogenic Leptospira (serovars Grippotyphosa, Pomona, Canicola, Hardjo, and Copehageni), Toxoplasma gondii, and Neospora caninum. The highest seroprevalence was detected for T. gondii (78%), followed by pathogenic Leptospira (14%). All the studied Leptospira serovars were confirmed in at least one animal. Two foxes seroconverted to Leptospira and one to T. gondii during the study period. No seroconversions were observed for the other pathogens. No risk factors, either intrinsic (sex, age) or extrinsic (season, year, and degree of landscape anthropization), were associated with the probability of being exposed to T. gondii. Our results indicate that T. gondii exposure is widespread in the Darwin's fox population, including in areas with minimal anthropization, and that T. gondii and pathogenic Leptospira might be neglected threats to the species. Further studies identifying the causes of morbidity and mortality in Darwin's fox are needed to determine if these or other pathogens are having individual or population-wide effects in this species.

Serological prevalence and risk factors of Toxoplasma gondii in Zoo Mammals in Chile.

Toxoplasma gondii is a zoonotic cosmopolitan protozoan that causes a high mortality rate among zoo mammals such as New World primates, meerkats, marsupials and Pallas' cat. It has been recently reported in Chile, mainly among wild populations, but also as the cause of death of a kangaroo and a mara. However, there has not been a T. gondii report at a Zoo population level in Chile in the last 35 years. The aim of the study was to estimate the seroprevalence and risk factors associated with T. gondii infection in mammals housed in a zoo located in the Metropolitan Region of Chile between 2011 and 2018. In this study, we analyzed 350 samples, from 324 animals, belonging to 57 species of carnivores, non-human primates, macropodids, ungulates and rodents to detect the presence of anti-T. gondii antibodies. Additionally, 20 animals were longitudinally sampled to evaluate intra-zoo infection. Using a commercial indirect Enzyme-Linked Immuno Sorbent Assay (ELISA) test, we detected T. gondii antibodies in 72 (22.2 %) samples. The overall seroprevalence estimates were 48.4 % in felines, 22.9 % in non-feline carnivores, 21.1 % in ungulates and 15.0 % in non-human primates. There were no positive samples from rodents or marsupials. Of animals sampled longitudinally, only a culpeo fox (Lycalopex cualpaeus) became seropositive along the study indicating exposition inside the facility. T. gondii seroprevalence differed significantly in taxonomic groups (p = 0.003), felines are statistically different from non-feline carnivores (NFC) (p = 0.040), ungulate (p = 0.027) and non-human primates (NHP) (p = 0.009). Annual prevalence comparison was performed showing no statistical difference (p = 0.941). A multivariable logistic regression was performed to ascertain the effect of taxonomic groups, proximity to water sources, diet, sex and type of housing on seropositivity. Only taxonomic group was statistically significant, indicating that NFC (OR = 0.35; 95 % CI = 0.15 - 0.83; p = 0.017), ungulates (OR = 0.30; 95 % CI = 0.13 - 0.69; p = 0.005), and NHP (OR = 0.25; 95 % CI = 0.09 - 0.72; p = 0.010) have lower risk of positivity to T. gondii compared to felines. Additionally, a black-faced spider monkey (Ateles chamek) and a siamang (Symphalangus syndactylus) were seropositive, being the first description of T. gondii infection in these species worldwide. As seen in previous studies, the widespread presence and exposure of T. gondii in zoo mammals was confirmed, and there may be contact with the agent and transmission within the zoo, which was confirmed by one animal became seropositive over the time. This fact could be a health problem for animals susceptible to fatal toxoplasmosis.