Ticks (Acari: Ixodidae) and rickettsiae associated with wild boars in a rural area of Minas Gerais, Brazil.

Wild boars or feral pigs are classified by the Brazilian Institute for the Environment and Renewable Resources (IBAMA) in "Category I of invasive exotic species". They cause economic losses, harm the environment, serve as hosts and reservoirs for several zoonotic disease agents, and provide a blood meal for tick species that act as vectors for zoonotic diseases. The objective of this study was to identify tick species on wild boars, assess host-seeking ticks in the related environment, and identify other potential tick hosts coexisting with wild boars on a farm located in the state of Minas Gerais, southeastern Brazil. Additionally, the study aimed to determine the presence of rickettsiae in these arthropods and assess the exposure of wild boars to rickettsiae species from the Spotted Fever Group and Rickettsia bellii through serology. A total of 3585 host-seeking ticks from three species (Amblyomma sculptum - 41.58%; Amblyomma dubitatum - 0.39% and Rhipicephalus microplus - 0.05%) were collected in the environment and A. sculptum was the most abundant species. Thirty-one wild boars were evaluated, resulting in the collection of 415 ticks, all of which were A. sculptum. Rickettsia DNA was not detected in samples of A. sculptum and R. microplus from the environment or in A. sculptum ticks from wild boars. However, all A. dubitatum ticks (n = 14) had Rickettsia bellii DNA confirmed by the species-specific PCR protocol. Out of the 31 serum samples from wild boars, 24 reacted with at least one Rickettsia antigen. Among these, seven individuals exhibited a reaction to a probable homologous antigen (PHA) of three rickettsiae species: R. rickettsii (n = 3), R. amblyommatis (n = 3) and R. rhipicephali (n = 1). Despite the high prevalence of seroreactivity, titers were low, indicating limited exposure to Rickettsia spp. Camera traps generated 874 animal records, capturing a total of 1688 individuals. At least 11 species of birds and 14 species of mammals (12 wild and two domestic) shared the environment with wild boars and potentially shared ticks with them. These findings provide baseline information for understanding the sharing of ticks and tick-borne pathogens between wild boars and other animals within the Cerrado biome. Further studies are necessary to monitor the potential and actual risk of wild boars to harbor infected ticks and their role in the transmission and maintenance cycle of Rickettsia spp.

Molecular identification of Sarcocystis neurona in tissues of wild boars (Sus scrofa) in the border region between Brazil and Uruguay.

Sarcocystis neurona, owing to its clinical importance in domestic animals, is currently one of the most studied agents, presenting a wide range of intermediate hosts that have not yet been described, mainly in wild fauna. Thus, the aim of this study was to describe the detection and molecular detection of S. neurona by amplification of the 18S rRNA region in the tissues of wild boars killed by boar control program in border Brazil Uruguay. A total of 79 samples of DNA from wild boar tissues from the LADOPAR/UFSM sampling bank were used, with Nested-PCR reactions being performed for amplification of the 18S rRNA region and the expected final product of 290 bp. Subsequently, the positive samples were subjected to restriction fragment length polymorphism (RFLP) technique with the restriction enzymes DdeI and HPAII. A second semi-Nested reaction was performed to obtain a larger sequence of nucleotides with amplification of the 18S region and the expected final product of 500 bp for S. neurona and Nested amplification ITS1 with product final of 367 pb. In 32 samples, it was possible to detect S. neurona both by nested Nested-PCR reaction and RFLP, and the presence of the agent was confirmed by sequencing, corresponding to 40.51% of the total tissues evaluated. This is the first report of the occurrence of this species of Sarcocystis in wild boars, and further studies evaluating the role of these animals as intermediate hosts, and in the epidemiology of this protozoan are necessary, as well as verifying the risk factors for infection.

Torque teno sus virus k2a (TTSuVk2a) in wild boars from northeastern Patagonia, Argentina.

Torque teno sus virus k2a (TTSuVk2a) is a member of the family Anelloviridae that can establish persistent infections in both domestic pigs and wild boars. Its association with diseases has not been precisely elucidated, and it is often considered only as a commensal virus. This infectious agent has been reported in herds throughout the world. In this study, we investigated the detection rate and diversity of TTSuVk2a in free-living wild boars from northeastern Patagonia, Argentina. Total DNA was extracted from tonsil samples of 50 animals, nested PCR assays were carried out, and infection was verified in 60% of the cases. Sequence analysis of the viral non-coding region revealed distinct phylogenetic groups. These clusters showed contrasting patterns of spatial distribution, which presented statistically significant differences when evaluating spatial aggregation. In turn, the sequences were compared with those available in the database to find that the clusters were distinguished by having similarity with TTSuVk2a variants of different geographic origin. The results suggested that Patagonian wild boar populations are bearers of diverse viral strains of Asian, European, and South American provenance.

One Health at gunpoint: Impact of wild boars as exotic species in Brazil - A review.

Wild boars have been listed among the 100 most invasive species worldwide, spreading impacts to all continents, with the exception of Antarctica. In Brazil, a major source of introduction was a commercial livestock importation for exotic meat market, followed by successive escapes and releases to natural ecosystems. Currently found in all six Brazilian biomes, with reports in 11 Brazilian states, wild boars have invaded natural and agricultural areas. Wild boars have been reportedly indicated as hosts and reservoirs of several zoonotic diseases in Brazil, including toxoplasmosis, salmonelosis, leptospirosis, brucellosis, tuberculosis, trichinellosis, and hepatitis E. Wild boars have been also associated with Brazilian spotted fever and rabies, infected while providing plentiful exotic blood supply for native ticks and hematophagous bats. Due to their phylogenetic proximity, wild boars may present ecological niche overlapping and direct disease risk to native white-lipped and collared peccaries. Moreover, wild boars may post an economical threat to Brazilian livestock industry due to restrictive diseases such as Aujeszky, enzootic pneumonia, neosporosis, hemoplasmosis, and classic swine fever. Finally, wild boars have directly impacted in environmentally protected areas, silting up water springs, rooting and wallowing native plants, decreasing native vegetal coverage, disbalancing of soil components, altering soil structure and composition. Wild boar hunting has failed as a control measure to date, according to the Brazilian Ministry of Environment, due to private hunting groups mostly targeting males, intentionally leaving females and piglets alive, disseminating wild boar populations nationwide. Meanwhile, non-government animal welfare organizations have pointed to animal cruelty of hunting dogs and wild boars (and native species) during hunting. Despite unanimous necessity of wild boar control, eradication and prevention, methods have been controversial and should focus on effective governmental measures instead occasional game hunting, which has negatively impacted native wildlife species while wild boars have continuously spread throughout Brazil.

Seropositive Wild Boars Suggesting the Occurrence of a Wild Cycle of Trichinella spp. in Brazil.

Trichinella is a zoonotic nematode traditionally detected worldwide in both domestic and wild animals. In South America, along with the occurrence of this parasite in domestic pigs and wild boars, there are reports of infection in wild carnivores. Brazil is considered free of the domestic cycle of Trichinella, but there is unpublished serological evidence of infection in wild boars, which changed the Brazilian status in OIE regarding the disease after an official communication. We investigated Trichinella spp. infection in wild boars and wild carnivores in the Southeastern region of Brazil. A total of 136 samples were tested, 121 from wild boars and 15 from wild carnivores. Artificial enzymatic digestion (AED) tests were performed on muscle samples from 37 wild boars and 15 wild carnivores, and 115 serum samples from wild boars were tested by iELISA. Seven serum samples from wild boars tested positive (7/115 = 6.1%, 95% CI 3.0-12.0), but no larvae were found in the AED. There was no significant difference between sex, age, and location of the samples. The serological results suggest that a wild cycle of Trichinella spp. may occur in Brazil, but further analyses should be performed to confirm the presence of the parasite.

Risk factors associated with ticks and Rickettsia spp. exposure in wild boars (Sus scrofa), hunting dogs, and hunters of Brazil.

BACKGROUND AND AIM: Wild boars have recently been implicated as the maintainers and carriers of Amblyomma spp. ticks, which are essential for Rickettsia spp. transmission. Consequently, wild boar hunting may increase the risk of tick exposure and subsequent human tick-borne infection and disease. Therefore, this study was conducted to evaluate the risk factors for ticks and Rickettsia spp. exposure in wild boars, hunting dogs, and hunters in Brazilian biomes. MATERIALS AND METHODS: The statistical relationship of Rickettsia spp. antibodies were evaluated using the Chi-square test in 80 wild boars, 170 hunting dogs, and 49 hunters. RESULTS: The only statistically significant difference in seropositivity found in this study was between male and female wild boars (p=0.034), probably associated with in-park exposure to Amblyomma brasiliense infected with Rickettsia spp. CONCLUSION: The absence of statistical differences in the associated risk factors for hunting dogs and hunters may indicate a random exposure to Rickettsia spp.

Molecular detection of Porcine cytomegalovirus (PCMV) in wild boars from Northeastern Patagonia, Argentina / Detección molecular de citomegalovirus porcino (PCMV) en jabalíes del noreste de la Patagonia argentina

ABSTRACT Porcine cytomegalovirus(PCMV) is a recognized pathogen of domestic swine that is widely distributed around the world. PCMV is the etiological agent of inclusion body rhinitis and has also been associated with other diseases that cause substantial losses in swine production. Wild boar populations can act as reservoirs of numerous infectious agents that affect pig livestock, including PCMV. The aim of this work was to assess the circulation of this virus in free-living wild boars that inhabit Northeastern Patagonia (Buenos Aires and Río Negro Provinces), Argentina. Nested-PCR assays were conducted to evaluate the presence of PCMV in samples of tonsil tissue collected from 62 wild boar individuals. It was found that the overall rate of infection was about 56%, with significant higher values (almost 90%) in the age group corresponding to piglets (animals less than 6 months old). In addition, a seasonal variation was observed in the PCMV detection rate, with an increase during the transition from summer to autumn. In conclusion, this study confirmed that wild boars are major carriers and dispersal agents of PCMV in Northeastern Patagonia, which raises the necessity to evaluate the extent to which this virus affects local livestock production.

RESUMEN El citomegalovirus porcino (CMVP) es un reconocido patógeno de los cerdos domésticos y cuenta con una amplia distribución mundial. Es el agente etiológico de la rinitis por cuerpos de inclusión y también se lo ha asociado con otras enfermedades que causan pérdidas sustanciales en la producción porcina. Las poblaciones de jabalíes pueden actuar como reservorios de numerosos agentes infecciosos que afectan al ganado porcino, incluido el CMVP. El objetivo de este trabajo fue evaluar la circulación de este virus en jabalíes de vida libre que habitan en la región noreste de la Patagonia argentina, en las provincias de Buenos Aires y Río Negro. Se realizaron ensayos de PCR anidada para evaluar la presencia de CMVP en muestras de tejido de amígdalas tomadas de 62 jabalíes. Se encontró que la tasa general de infección fue de aproximadamente el 56%, con valores significativamente más altos (casi el 90%) en el grupo de edad correspondiente a los lechones (animales con menos de 6meses). Además, se observó una variación estacional en la tasa de detección de CMVP, con un incremento durante la transición de verano a otoño. En conclusión, este estudio confirmó que los jabalíes son importantes portadores y agentes de dispersión del CMVP en el noreste patagónico, lo cual plantea la necesidad de evaluar en qué medida este virus afecta la producción ganadera local.

Risk of Introduction of Classical Swine Fever Into the State of Mato Grosso, Brazil.

Classical swine fever (CSF) is considered one of the most important diseases of swine because of the far-reaching economic impact the disease causes to affected countries and regions. The state of Mato Grosso (MT) is part of Brazil's CSF-free zone. CSF status is uncertain in some of MT's neighboring States and countries, which has resulted in the perception that MT is at high risk for the disease. However, the risk for CSF introduction into MT has not been previously assessed. Here, we estimated that the risk for CSF introduction into the MT is highly heterogeneous. The risk associated with shipment of commercial pigs was concentrated in specific municipalities with intense commercial pig production, whereas the risk associated with movement of wild boars was clustered in certain municipalities located close to the state's borders, mostly in northern and southwestern MT. Considering the two pathways of possible introduction assessed here, these results demonstrate the importance of using alternative strategies for surveillance that target different routes and account for different likelihoods of introduction. These results will help to design, implement, and monitor surveillance activities for sustaining the CSF-free status of MT at times when Brazil plans to expand the recognition of disease-free status for other regions in the country.

EXPOSURE OF WILD BOAR (SUS SCROFA) TO THE COMMON VAMPIRE BAT AND LACK OF IMMUNE PROTECTION TO RABIES VIRUS IN BRAZILIAN HUNTERS.

Rabies virus is recognized as one of the most fatal zoonotic agents affecting all mammals. Wild boars (Sus scrofa), classified as a large-size exotic invasive species in Brazil with nationwide hunting permitted, may serve as an extra blood source for the common vampire bat (Desmodus rotundus). Our aim was to document wild boar exposure to vampire bats to determine the seroprevalence of rabies virus antibodies in wild boars and to determine the immune status of hunters in southern and central-western Brazilian regions. Serum samples were collected from 80 wild boars and 49 hunters from natural and degraded areas of the Atlantic Forest biome of southern Brazil and in degraded areas of the Cerrado biome of central-western Brazil. The rabies-modified rapid fluorescent focus inhibition test was performed to detect the presence of rabies virus neutralizing antibodies in wild boars and considered seropositive when ≥0.10 IU/mL. The simplified fluorescence inhibition microtest was used for samples from hunters with a titer of ≥0.50 IU/mL and considered indicative of seroconversion. While 11% (9/80) of wild boars had serum titers for rabies exposure (≥0.10 IU/mL), 88% (43/49) of corresponding hunters lacked immune protective titers (<0.50 IU/mL). Wild boars showed serum titers for rabies likely due to contact with contaminated saliva of vampire bats or from infected carcass consumption. Additionally, Brazilian wild boars can be exposed to rabies and may play an important role in the sylvatic rabies cycle by providing a blood supply for vampire bats, highlighting the possibility of direct transmission of rabies virus to hunting dogs and hunters. These findings suggested hunters are a potential risk group for contracting rabies, and the World Health Organization may consider adding this occupation to their recommendations of who should receive the pre-exposure rabies vaccination.

Molecular detection of Porcine cytomegalovirus (PCMV) in wild boars from Northeastern Patagonia, Argentina.

Porcine cytomegalovirus (PCMV) is a recognized pathogen of domestic swine that is widely distributed around the world. PCMV is the etiological agent of inclusion body rhinitis and has also been associated with other diseases that cause substantial losses in swine production. Wild boar populations can act as reservoirs of numerous infectious agents that affect pig livestock, including PCMV. The aim of this work was to assess the circulation of this virus in free-living wild boars that inhabit Northeastern Patagonia (Buenos Aires and Río Negro Provinces), Argentina. Nested-PCR assays were conducted to evaluate the presence of PCMV in samples of tonsil tissue collected from 62 wild boar individuals. It was found that the overall rate of infection was about 56%, with significant higher values (almost 90%) in the age group corresponding to piglets (animals less than 6 months old). In addition, a seasonal variation was observed in the PCMV detection rate, with an increase during the transition from summer to autumn. In conclusion, this study confirmed that wild boars are major carriers and dispersal agents of PCMV in Northeastern Patagonia, which raises the necessity to evaluate the extent to which this virus affects local livestock production.

Molecular identification of Mycobacterium spp. isolated from Brazilian wild boars.

Wild boars (Sus scrofa) are susceptible to mycobacterial infections, including tuberculous and non-tuberculous mycobacteria. Recently, Mycobacterium spp. infections were described in Brazilian wild boars, which can act as bacterial reservoirs. Here, we aim to characterize 15 Mycobacterium spp. isolates from Brazilian wild boars' tissues through partial sequencing of the heat shock protein 65 (hsp65) gene and phylogenetic analysis. The isolates were classified as M. tuberculosis (33.3%), M. colombiense (33.3%), M. avium subsp. hominissuis (13.3%), M. parmense (13.3%) and M. mantenii (6.66%). The isolates classified as M. tuberculosis were confirmed as variant bovis by PCR. At phylogenetic analysis some isolates formed separated clades, indicating genetic variability. Different Mycobacterium species were recovered from wild boars circulating in Brazil, including mycobacteria associated to zoonotic infections, such as M. tuberculosis. In addition, this is the first report in Brazilian wild boars on M. mantenii and M. parmense detection, two recently described pathogenic mycobacteria. However, the isolates' genetic diversity-i.e. identities lower than 100% when compared to reference sequences-suggests that other genotyping tools would allow a deeper characterization. Nonetheless, the reported data contributes to the knowledge on mycobacterial infections in wild boars from Brazil.

Health profile of free-range wild boar (Sus scrofa) subpopulations hunted in Santa Catarina State, Brazil.

Wild boars represent a potential dissemination source of important pathogens to public and animal health, since they can host several pathogens. This study aimed to evaluate the health profile of wild boars hunted for population control in the state of Santa Catarina (SC), south of Brazil. For this study, tissue and blood samples were collected from 61 wild boars hunted from October 2017 to November 2018. All 61 serum samples were screened for antibodies by different assays, presenting the following seroprevalence: 52.4% for porcine circovirus type 2 (PCV2); 21.3% for Leptospira spp.; 19.7% for Mycoplasma hyopneumoniae; 13.1% for hepatitis E virus (HEV); and 9.8% for influenza A virus (IAV). No seroreaction was observed for Brucella spp. and classical swine fever virus (CSFV). At necropsy, the main lesions observed were evidence of pulmonary metastrongylid parasites, haemorrhagic lymph nodes and white-spotted liver. Histopathological analyses mainly showed interstitial pneumonia, bronchopneumonia and bronchi-associated lymphoid tissue (BALT) hyperplasia. There was a significant association between metastrongylid parasitism and BALT hyperplasia in lungs. Pathological findings of wild boars indicate that the assessed individuals were not exposed to notifiable diseases, such as CSFV, African swine fever virus and foot-and-mouth disease, which cause economic damage by non-tariff trade barriers. However, the detection of antibodies against PCV2 and M. hyopneumoniae, pathogens that impact commercial production, indicates the circulation of these agents, with frequency variation in wild populations. This shows the potential risk of pathogen transmission among domestic swine populations and free-living ones. The detection of antibodies against zoonotic agents such as HEV, IAV and Leptospira spp., other than characterizing the pathogens' circulation in these populations, suggests a potential risk to public health and pig farming, depending on the interactions established with the environment, humans and domestic swine.

Study of Trichinella patagoniensis in wild boars.

Trichinellosis is a zoonotic disease, which represents a significant public health concern in some South American countries, such as Argentina and Chile. Its impact is essentially due to absence of adequate control measures on meat from game animals, as well as the presence of illegal slaughterhouses and the trade of meat products without being tested for this parasite. In Argentina, trichinellosis is an endemic disease. At present, Trichinella spiralis, Trichinella patagoniensis, Trichinella pseudospiralis, and Trichinella britovi have been detected in animals from Argentina. Until now, T. patagoniensis had only been found in mountain cougars (Puma concolor) in Argentina but there is limited information available. The present study intends to determine susceptibility, serological response and distribution of muscle larvae in wild boars infected with T. patagoniensis, T. spiralis and T. pseudospiralis. For each of the Trichinella species five wild boars were inoculated with 20,000 muscle larvae. Except for two specimens which died during the experiment, the animals were euthanized 19 weeks post infection (pi). Blood samples were collected throughout the study in order to determine the antibody kinetics. Also, nine muscle samples from each specimen were taken and analysed for determination of larval distribution. Additionally, four muscle samples were used to obtain muscle juices. Wild boars infected with T. patagoniensis showed little to no larvae in the muscle samples analysed while animals infected with T. spiralis and T. pseudospiralis had a significantly high larval load in all the samples analysed. Optical density (OD) values remained above the cut-off value throughout the experiment. This is the first study to characterize the biological aspects of T. patagoniensis in wild boars.

Complete genome characterization of porcine circovirus 3 recovered from wild boars in Southern Brazil.

In the present study, the complete nucleotide sequence of porcine circovirus 3 (PCV3) recovered from wild boars lymph nodes is described. The full genome was named PCV3-wb/Br/RS and comprises 2,000 nucleotides with two open reading frames (ORFs) with a stem-loop motif in intergenic region. The ORFs are oriented in opposite directions and encode the putative capsid (Cap) and replicase (Rep) proteins. Based on amino acid motif analysis, PCV3-wb/Br/RS as well as most of the sequences from wild boars are classified as PCV3b. Phylogenetic analysis including 97 PCV3 sequences available in databases showed that the PCV3-wb/Br/RS genome is more closely related to genomes recovered in Spain, China, Germany and Denmark. Phylogenetic inferences among PCV3-wb/Br/RS and other circoviruses confirmed that these seem to have a most recent common ancestor with bat-associated circoviruses. In addition, PCV3 infection was investigated by real-time PCR in a cohort of 80 wild boars in Southern Brazil. A total of 29 animals (36.3%) were PCV3-positive leading the conclusion that PCV3 is circulating in the wild boar population in Southern Brazil. The role played by PCV3-like infections in wild boars and the risk these could pose to commercial swine production within that region remains to be further investigated.

Achievements and Challenges of Classical Swine Fever Eradication in Brazil.

Classical swine fever virus (CSFV) causes one of the most critical diseases in the porcine industry worldwide. In Brazil, the first description of the infection was reported in 1888, and the national recognition of the first free zone (FZ) occurred in 2001. Brazil has been recently recognized (2015-2016) by the World Organisation for Animal Health (OIE) with an FZ involving 15 states and the Federal District, corresponding to 95% of the industrial production of pigs in the country, and a non-free zone (NFZ), comprised by the North and Northeast regions of the country, with approximately 18% of the national pig herd and 5% of industrial production. This review aims to describe the history, the control and eradication actions, the recent occurrence of outbreaks in the NFZ, and the results obtained by the surveillance systems' action in the FZ for CSF in Brazil since its creation. In the passive surveillance system, the notification of the suspect cases of classical swine fever (CSF) is mandatory while in the active surveillance system adopted in the FZ consists of serological monitoring of certified swine breeding farms (CSBFs), intensive pig farming (IPF), non-technified pig herds (NTPig), surveillance in slaughterhouses and monitoring the populations of wild pigs. In this region, the last outbreaks of the disease occurred in 1998, while in the NFZ, 28 outbreaks were detected from 2005 to 2017, with an apparent lethality rate of 93.96% (840/894). However, in 2018 and 2019, 68 new outbreaks were registered with an apparent lethality rate of 75.05% (1095/1459). Therefore, in 2019, the Brazil CSF-Free Strategic Plan was created to eradicate the infection from the country's NFZ, since outbreaks in this region present a risk of reintroducing the disease FZ. Finally, differences in characteristics between the regions show factors that still need to be considered for the construction of a robust surveillance system in the NFZ and some improvements in the FZ. Thus, the control of CSF throughout the Brazilian territory requires strict sanitary guidelines, promoting animal health and, consequently, the national production chain's competitiveness.

New pieces in the transmission cycle of the hepatitis E virus in South America: first viral detection in wild boars from Argentina.

BACKGROUND: The hepatitis E virus (HEV) causes hepatitis worldwide. In Europe, wild boars are known to be viral reservoirs and sources of infection. In South America, there is a lack of information about HEV in these animals. METHODS: A total of 102 wild boar serum samples from Argentina (2014-2017) were studied for serological and molecular HEV detection. RESULTS: We obtained a seroprevalence of 19.6%, similar to that recently described in Uruguay (the only antecedent in South America). HEV ribonucleic acid (RNA) was amplified in two anti-HEV-positive samples. CONCLUSIONS: This is the first report of HEV circulation in wild boars from Argentina, adding evidence to the findings obtained in Uruguay indicating that wild boars could be viral reservoirs in South America.

Trichinella spiralis Infecting Wild Boars in Southern Chile: Evidence of an Underrated Risk.

Trichinellosis in Chilean wild animals has scarcely been documented. The introduction of wild boars into the wild environment represents a viable new host with a potential risk of infection for human health. The aim of this study was to determine the presence and prevalence of Trichinella in wild boars. Two hundred seventy eight wild boars from of the Southern Chile were examined by compression and artificial digestion techniques. The larvae in the positive samples were collected for taxonomic analysis through polymerase chain reaction-inter-simple sequence repeats and to calculate the parasitic burden. A prevalence of 1.8% (5/278) of infected animals and an average parasitic burden of 6.8 ± 2.1 larvae per gram were estimated. The only species identified by molecular techniques was Trichinella spiralis. Prevalence of T. spiralis in wild boars was similar to those described around the world. T. spiralis infection rate and parasite burden detected in Chilean wild boars represent a certain food-borne risk for human population.

Serological and virological survey of hepatitis E virus (HEV) in animal reservoirs from Uruguay reveals elevated prevalences and a very close phylogenetic relationship between swine and human strains.

Hepatitis E virus (HEV) infection is an issue of public health concern in high-income and non-endemic countries. Increasing evidence supports the hypothesis of a zoonotic route as the main mode of infection in this epidemiological setting, since the transmission of genotypes HEV-3 and HEV-4 from reservoirs to humans has been demonstrated. In America, studies have confirmed the circulation of HEV in pig herds but the zoonotic role of wild boars has never been evaluated. Uruguay has a high burden of HEV- associated acute hepatitis, and a close phylogenetic relationship was observed among human HEV-3 strains and European isolates detected in swine. However in this context, swine herds have never been surveyed. Herein is reported a survey of HEV in swine herds, pigs at slaughter-house and free-living wild boar populations. Two-hundred and twenty sera and 150 liver tissue samples from domestic pigs, and 140 sera from wild boars were tested for HEV by ELISA and PCR-based approaches. All tested swine farms resulted seropositive with an overall rate of 46.8%. In turn, 22.1% of the wild boars had anti-HEV antibodies. HEV RNA was detected in 16.6% and 9.3% of liver samples from slaughter-age pigs and adult wild boars sera, respectively. Three strains from domestic pig were also amplified by nested-PCR approaches. By contrast, none of the positive samples obtained from wild boars could be confirmed by nested-PCR. Phylogenetic analysis revealed a very high nucleotide identity among swine strains and sequences obtained from humans in Uruguay. Results showed that HEV is widely distributed among swine herds in Uruguay. Additionally, this study evidences for the first time in the American continent that wild boar populations are a reservoir for HEV, though its zoonotic role remains to be elucidated. Altogether, data presented here suggest a high zoonotic risk of HEV transmission from swine to humans.

Molecular survey of porcine teschovirus, porcine sapelovirus, and enterovirus G in captive wild boars (Sus scrofa scrofa) of Paraná state, Brazil / Estudo molecular de teschovírus suíno, sapelovírus suíno e enterovírus G em javalis (Sus scrofa scrofa) de cativeiro no Paraná

Porcine teschovirus (PTV), porcine sapelovirus (PSV), and enterovirus G (EV-G) are infectious agents specific to pig host species that are endemically spread worldwide. This study aimed to investigate the natural infection by these porcine enteric picornaviruses in wild boars (Sus scrofa scrofa) of Paraná state, Brazil, and to evaluate peccaries (Pecari tajacu and Tayassu pecari) as alternative host species for these viruses. Fecal samples (n=36) from asymptomatic wild boars (n=22) with ages ranging from 2 to 7 months old (young, n=14) and 2 to 4 years old (adult, n=8) and from peccaries (6 to 8 months old, n=14) were collected from a farm and a zoo, respectively, both located in Paraná state. Reverse transcription-polymerase chain reaction (RT-PCR) and nested-PCR (n-PCR) assays targeting the 5'non-translated region of the virus genome were used for screening the viruses. Porcine enteric picornaviruses were detected in 12 out of the 22 wild boar fecal samples. According to each of the viruses, EV-G was most frequently (11/22, 50%) detected, followed by PTV (10/22, 45.5%) and PSV (4/22, 18.2%). Regarding the age groups, young wild boars were more frequently (9/14, 64.3%) infected with PTV, PSV, and EV-G than adult animals (3/8, 37.4%). One n-PCR amplified product for each of the viruses was submitted to sequencing analysis and the nucleotide sequences were compared with the related viruses, which showed similarities varying from 97.7% to 100% for PTV, 92.4% to 96.2% for PSV, and 87.1% to 100% for EV-G. Peccaries tested negative for the viruses and in this study they did not represent infection reservoirs. This study is the first to report the molecular detection of PTV, PSV, and EV-G from captive wild boars in a South American country and the first to screen peccaries as alternative host species for porcine enteric picornavirus.(AU)

Teschovírus suíno (PTV), sapelovírus suíno (PSV) e enterovírus G(EV-G) são agentes infecciosos específicos da espécie suína que estão endemicamente disseminados em todo o mundo. O objetivo deste estudo foi investigar a infecção natural por estes picornavírus entéricos suínos em javalis (Sus scrofa scrofa) do estado do Paraná, Brasil e avaliar pecaris (Pecari tajacu e Tayassu pecari) como hospedeiros alternativos para estes vírus. Amostras fecais (n=36) de javalis assintomáticos (n=22) com idades de 2 a 7 meses (jovens, n=14) e 2 a 4 anos (adultos, n=8) e de pecaris (6 a 8 meses de idade, n=14) foram coletadas em um cativeiro e zoológico, respectivamente, ambos localizados no estado do Paraná. A transcrição reversa seguida por reações da polimerase em cadeia (RT-PCR) e nested-PCR com alvo na região 5'-não traduzida do genoma viral foram utilizadas para a identificação dos vírus. Picornavírus entéricos suínos foram detectados em 12 das 22 amostras fecais de javalis. De acordo com cada um dos vírus, EV-G foi mais frequentemente (11/22, 50%) detectado, seguido pelo PTV (10/22; 45,5%) e PSV (4/22; 18,2%). Considerando os grupos de idade, javalis jovens foram mais frequentemente (9/14; 64,3%) infectados com PTV, PSV e EV-G do que os javalis adultos (3/8; 37,4%). Um produto amplificado na nested-PCR para cada um dos vírus foi submetido à análise de sequenciamento e as sequências de nucleotídeos foram comparadas com vírus relacionados, o que mostrou que as similaridades variaram entre 97,7% a 100% para o PTV, 92,4% a 96,2% para o PSV e 87,1% a 100% para o EV-G. Os pecaris foram negativos para as viroses investigadas e neste estudo não se apresentaram como hospedeiros alternativos para as infecções. Este estudo é o primeiro a relatar a detecção molecular de PTV, PSV e EV-G em javalis de cativeiro de um país da América Latina e o primeiro a avaliar pecaris como espécie hospedeira alternativa para picornavírus entéricos suínos.(AU)

Molecular survey of porcine teschovirus, porcine sapelovirus, and enterovirus G in captive wild boars (Sus scrofa scrofa) of Paraná state, Brazil / Estudo molecular de teschovírus suíno, sapelovírus suíno e enterovírus G em javalis (Sus scrofa scrofa) de cativeiro no Paraná

Porcine teschovirus (PTV), porcine sapelovirus (PSV), and enterovirus G (EV-G) are infectious agents specific to pig host species that are endemically spread worldwide. This study aimed to investigate the natural infection by these porcine enteric picornaviruses in wild boars (Sus scrofa scrofa) of Paraná state, Brazil, and to evaluate peccaries (Pecari tajacu and Tayassu pecari) as alternative host species for these viruses. Fecal samples (n=36) from asymptomatic wild boars (n=22) with ages ranging from 2 to 7 months old (young, n=14) and 2 to 4 years old (adult, n=8) and from peccaries (6 to 8 months old, n=14) were collected from a farm and a zoo, respectively, both located in Paraná state. Reverse transcription-polymerase chain reaction (RT-PCR) and nested-PCR (n-PCR) assays targeting the 5'non-translated region of the virus genome were used for screening the viruses. Porcine enteric picornaviruses were detected in 12 out of the 22 wild boar fecal samples. According to each of the viruses, EV-G was most frequently (11/22, 50%) detected, followed by PTV (10/22, 45.5%) and PSV (4/22, 18.2%). Regarding the age groups, young wild boars were more frequently (9/14, 64.3%) infected with PTV, PSV, and EV-G than adult animals (3/8, 37.4%). One n-PCR amplified product for each of the viruses was submitted to sequencing analysis and the nucleotide sequences were compared with the related viruses, which showed similarities varying from 97.7% to 100% for PTV, 92.4% to 96.2% for PSV, and 87.1% to 100% for EV-G. Peccaries tested negative for the viruses and in this study they did not represent infection reservoirs. This study is the first to report the molecular detection of PTV, PSV, and EV-G from captive wild boars in a South American country and the first to screen peccaries as alternative host species for porcine enteric picornavirus.

Teschovírus suíno (PTV), sapelovírus suíno (PSV) e enterovírus G(EV-G) são agentes infecciosos específicos da espécie suína que estão endemicamente disseminados em todo o mundo. O objetivo deste estudo foi investigar a infecção natural por estes picornavírus entéricos suínos em javalis (Sus scrofa scrofa) do estado do Paraná, Brasil e avaliar pecaris (Pecari tajacu e Tayassu pecari) como hospedeiros alternativos para estes vírus. Amostras fecais (n=36) de javalis assintomáticos (n=22) com idades de 2 a 7 meses (jovens, n=14) e 2 a 4 anos (adultos, n=8) e de pecaris (6 a 8 meses de idade, n=14) foram coletadas em um cativeiro e zoológico, respectivamente, ambos localizados no estado do Paraná. A transcrição reversa seguida por reações da polimerase em cadeia (RT-PCR) e nested-PCR com alvo na região 5'-não traduzida do genoma viral foram utilizadas para a identificação dos vírus. Picornavírus entéricos suínos foram detectados em 12 das 22 amostras fecais de javalis. De acordo com cada um dos vírus, EV-G foi mais frequentemente (11/22, 50%) detectado, seguido pelo PTV (10/22; 45,5%) e PSV (4/22; 18,2%). Considerando os grupos de idade, javalis jovens foram mais frequentemente (9/14; 64,3%) infectados com PTV, PSV e EV-G do que os javalis adultos (3/8; 37,4%). Um produto amplificado na nested-PCR para cada um dos vírus foi submetido à análise de sequenciamento e as sequências de nucleotídeos foram comparadas com vírus relacionados, o que mostrou que as similaridades variaram entre 97,7% a 100% para o PTV, 92,4% a 96,2% para o PSV e 87,1% a 100% para o EV-G. Os pecaris foram negativos para as viroses investigadas e neste estudo não se apresentaram como hospedeiros alternativos para as infecções. Este estudo é o primeiro a relatar a detecção molecular de PTV, PSV e EV-G em javalis de cativeiro de um país da América Latina e o primeiro a avaliar pecaris como espécie hospedeira alternativa para picornavírus entéricos suínos.