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Host genetic variability can modulate infection resistance, although its role in infection clearance remains unclear. Hookworm disease ( Uncinaria sp.) is the leading cause of pup mortality in several otariid species, although the parasite can be cleared through immune-mediated processes. We evaluated the association of host genetic diversity, body condition and immune response with hookworm resistance and/or clearance in the South American fur seal ( Arctocephalus australis ). Uninfected pups had higher heterozygosity than parasitized individuals, indicating a negative relationship between heterozygosity and the chances of infection. Likewise, pups that died of hookworm infection had lower heterozygosity than those that died of non-infectious causes. Interestingly, once infected, pups that survived hookworm infection had heterozygosities similar to pups that died of hookworm disease. However, pups that cleared the infection had a higher body mass and parasite-specific immunoglobulin G levels than those that did not recover or died of hookworm disease. Thus, although heterozygosity predicted resistance to and mortality from hookworm infections, it did not affect parasite clearance, which was facilitated by better body condition and adaptive immune responses. This demonstrates that host genetic variability and host-environment interactions influence disease dynamics, acting at different, well-defined stages of infection.

Wild rodents often harbor Cryptosporidium species that can be transmitted to multiple mammal hosts. In Chile, little is known about Cryptosporidium in wild rodents, and available studies have been focused on morphological findings with no molecular-based evidence. A longitudinal survey was conducted between 2021 and 2022 to investigate the occurrence of Cryptosporidium spp. in populations of the Darwin's leaf-eared mouse ( Phyllotis darwini ) living in protected and rural transitional areas in north-central Chile, using staining and molecular methods. A total of 247 fecal samples were collected and examined by the modified Ziehl-Neelsen (ZN) staining test, 54 of which were positive for Cryptosporidium -like oocysts. Molecular analyses were carried out by PCR of the partial 18S ribosomal RNA and 60 kDa glycoprotein ( gp60 ) genes. Cryptosporidium infection was confirmed in 34 samples (13.7 %) based on the PCR amplification, and individual (i.e., sex, and body mass index) and ecological variables (i.e., type of site and season) were not statistically significant (p > 0.05). Using the nucleotide sequencing of the partial 18S rRNA gene, Cryptosporidium parvum was identified in nine isolates. Also, C. parvum subgenotype family IIa was determined in seven samples by the partial gp60 gene, including the subtype IIaA17G4R1 in two samples . This is the first molecular evidence of Cryptosporidium parvum IIa in Phyllotis darwini in Chile. These results indicate potential cross-species transmition between wild rodents and domestic-wild animals in north-central Chile. More research is needed to understand better the role of wild rodents in the transmission of Cryptosporidium spp. in Chile., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Gerardo Acosta-Jamett reports financial support was provided by ANID Fondecyt Regular 2021 N. 1211190. Patricio D. Carrera-Jativa reports financial support was provided by ANID Programa de Becas Doctorado Nacional N. 21200220. Patricio D. Carrera-Jativa reports financial support was provided by WWF Russell E. Train Fellowship. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Mammals in drylands face environmental challenges exacerbated by climate change. Currently, human activity significantly impacts these environments, and its effects on the energy demands experienced by individuals have not yet been determined. Energy demand in organisms is managed through elevations in glucocorticoid levels, which also vary with developmental and health states. Here, we assessed how anthropization, individual characteristics, and seasonality influence hair glucocorticoid concentration in the Darwin's leaf-eared mouse ( Phyllotis darwini ) inhabiting two areas with contrasting anthropogenic intervention in a semi-arid ecosystem of northern Chile. Hair samples were collected (n = 199) to quantify hair corticosterone concentration (HCC) using enzyme immunoassays; additionally, sex, body condition, and ectoparasite load were recorded. There were no differences in HCC between anthropized areas and areas protected from human disturbance; however, higher concentrations were recorded in females, and seasonal fluctuations were experienced by males. The results indicate that animals inhabiting semi-arid ecosystems are differentially stressed depending on their sex. Additionally, sex and season have a greater impact on corticosterone concentration than anthropogenic perturbation, possibly including temporal factors, precipitation, and primary production. The influence of sex and seasonality on HCC in P. darwini make it necessary to include these variables in future stress assessments of this species.

The tick Amblyomma tigrinum inhabits areas with diverse climatic conditions, with adult stages parasitizing wild canids, such as chilla (Lycalopex griseus) and culpeo (Lycalopex culpaeus) foxes. We described the infestation loads in wild foxes captured at three sites (periurban, rural and wild) through an anthropization gradient in north-central Chile. We tested whether local-scale environmental and/or individual host factors can predict tick abundance by using negative binomial models. During 2018-2020 (spring and summer), we captured 116 foxes (44 chillas and 72 culpeos), and 102 of them were infested with ticks (87.9%, CI=80.6-93.2%). We collected 996 A. tigrinum adult ticks, estimating a total mean abundance of 8.6±0.8 ticks/host. Periurban and rural foxes harbored greater tick loads than foxes from the wild site (2.34 and 1.71 greater, respectively) while tick abundance in summer decreased by up to 57% compared to spring. Tempered, more humid climate conditions of the periurban site could favor the development and survival of adults A. tigrinum; and ticks may have adopted a quiescent stage or similar survival mechanisms to cope with summer temperature increases related to the ongoing megadrought. Further studies are warranted to understand the underlying factors determining the life cycle of A. tigrinum at larger spatiotemporal scales.

Competing Interests: All authors have completed and submitted the International Committee of Medical Journal Editors form for disclosure of potential conflicts of interest. Thomas Weitzel, Constanza Martínez-Valdebenito, Katia Abarca, and Gerardo Acosta-Jamett report institutional support (including equipment and laboratory materials for diagnostic testing), travel support, and honoraria from the National Research and Development Agency of Chile (Agencia Nacional de Investigación y Desarrollo) and the National Fund for Scientific and Technological Development (Fondo Nacional de Desarrollo Científico y Tecnológico). No other potential conflicts of interest were disclosed.

Despite the extensive information on the effects of habitat alteration on the structure of helminth communities in small mammals, the evidence is still inconclusive. A systematic review was carried out using the PRISMA (Preferred Reporting Items for Systematic Review and Meta-Analysis) guideline to compile and synthesize available literature on the influence of habitat alteration on the structure of helminth communities in small mammals. The aim of this review was to describe the variation in infection rates of helminth species associated with habitat alteration and to discuss the theoretical framework that may explain such changes in relation to parasite, host, and environmental features. Twenty-three scientific articles published between 2005 and 2022 were reviewed, 22 of which investigated parasite prevalence, 10 parasite burden, and 14 parasite richness in both altered and natural habitats. Information in assessed articles suggests that the structure of helminth communities in small mammals can be impacted by anthropogenic habitat alteration in various ways. Infection rates of monoxenous and heteroxenous helminths may increase or decrease in small mammals depending on whether their hosts (definitive and intermediate) are available, and environmental and host conditions modify the survival and transmission of parasitic forms. Also, given that habitat alteration may favor inter-species contacts, transmission rates of low host-specific helminths could be increased due to exposure to new reservoir hosts. In a continually changing world, it is essential to assess the spatio-temporal variations of helminth communities in wildlife inhabiting altered and natural habitats to determine potential impacts on wildlife conservation and public health., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The main objective of this study was to estimate the performance, under local epidemiological conditions, of two in-house ELISA assays for the combined detection of anti-SARS-CoV-2 IgA, IgM, and IgG immunoglobulins. A total of 94 serum samples were used for the assessment, where 44 corresponded to sera collected before the pandemic (free of SARS-CoV-2 antibodies), and 50 sera were collected from confirmed COVID-19 patients admitted to the main public hospital in the city of Valdivia, southern Chile. The Nucleocapsid (Np) and the receptor-binding domain (RBD) proteins were separately used as antigens (Np and RBD ELISA, respectively) to assess their diagnostic performance. A receiver operating characteristic (ROC) analysis was performed to estimate the optical density (OD) cut-off that maximized the sensitivity (Se) and specificity (Sp) of the ELISA assays. Np ELISA had a mean Se of 94% (95% CI = 83.5-98.8%) and a mean Sp of 100% (95% CI = 92.0-100%), with an OD 450 nm positive cut-off value of 0.88. On the other hand, RBD ELISA presented a mean Se of 96% (95% CI = 86.3-99.5%) and a mean Sp of 90% (95% CI = 78.3-97.5%), with an OD 450 nm positive cut off value of 0.996. Non-significant differences were observed between the Se distributions of Np and RBD ELISAs, but the latter presented a significant lower Sp than Np ELISA. In parallel, collected sera were also analyzed using a commercial lateral flow chromatographic immunoassay (LFCI), to compare the performance of the in-house ELISA assays against a commercial test. The LFCI had a mean sensitivity of 94% (95% CI = 87.4-100%) and a mean specificity of 100% (95% CI = 100-100%). When compared to Np ELISA, non-significant differences were observed on the performance distributions. Conversely, RBD ELISA had a significant lower Sp than the LFCI. Although, Np ELISA presented a similar performance to the commercial test, this was 2.5 times cheaper than the LFCI assay (labor cost not considered). Thus, the in-house Np ELISA could be a suitable alternative tool, in resource limited environments, for the surveillance of SARS-CoV-2 infection, supporting further epidemiological studies., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Ramírez-Reveco et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Scrub typhus is a potentially severe infection caused by bacteria of the genus Orientia, endemic in Asia-Pacific and recently discovered in southern Chile. The presented study aimed to determine the prevalence and species richness of rodent-associated trombiculid mites and their infection with Orientia spp. in different areas of two regions in southern Chile., Methodology/principal Findings: During summer 2020, trombiculid mites were collected from rodents captured in three areas in southern Chile known to be endemic for scrub typhus (Cochamó and Chiloé Island in the Los Lagos Region and Tortel in the Aysén Region). A total of 132 rodents belonging to five species were captured using Sherman-like traps; 89.4% were infested with trombiculids. Mite specimens were morphologically identified and subsequently tested by Orientia-specific qPCR. Six mite species were identified. Among chigger-infested rodents, 33.9% carried Orientia-positive mites; this rate was higher in Tortel (63.8%) than in Cochamó (45.0%) and Chiloé Island (2.0%). The analysis of individual mites (n = 901) revealed that 31.2% of Herpetacarus antarctica samples (n = 202) were positive for Orientia DNA; the prevalence was 7.0% in Paratrombicula neuquenensis (n = 213), 6.9% in Herpetacarus eloisae (n = 144), 3.6% in Argentinacarus expansus (n = 55), and 0% in Paratrombicula goffi (n = 110) and Quadraseta chiloensis (n = 177). The southernmost site (Tortel) showed the highest rates of trombiculid infestation, trombiculid load, and Orientia infection in the captured rodents., Conclusions/significance: Our study provides new insights into the trombiculid fauna and prevalence of Orientia in mites collected from wild rodents in southern Chile. Orientia DNA was detected in four of the six mite species. Rates of infestation, mite loads, and Orientia prevalences differed geographically and were highest in the Aysén Region. Our data improve our knowledge on possible vectors of scrub typhus and their distribution in Chile., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Silva de la Fuente et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Gastrointestinal parasites are well-documented in small mammals from north-central Chile, but little is known about endoparasites of rodents in southern Chile. A survey was conducted between January and February 2018 to evaluate gastrointestinal parasites and risk factors of wild rodents that live in rural areas in Northern Chiloé Island, Chile. A total of 174 fecal samples from rodents of six native and one introduced species were collected and examined using the Mini-FLOTAC method. Also, 41 individuals of four native wild rodent species were examined furtherly to determinate adult parasites from gastrointestinal tracts. The overall prevalence of endoparasites was 89.65% (156). Helminth egg types included: Rodentolepis spp., Capillariidae, Trichuris sp., Syphacia sp., oxyurid-type eggs, Strongyloides sp., Spirurid-type eggs, Strongilid-type eggs, Moniliformis sp., and an unidentified nematode egg and larvae. Protozoa comprised coccidia, amoeba, and unidentified cysts. From necropsies, adult parasites involved Syphacia sp. Trichuris sp., Protospirura sp. and Physaloptera sp. In Abrothrix olivacea, individuals with low-body-mass index exhibited reduced infection probability for Spirurid-type and Strongilid-type eggs. Some parasites in this study may affect human health. In rural settings where environmental conditions are changing, more research should be undertaken to understand parasitic infections in wildlife and implications for public health and conservation.

The exposure of a research team to chigger mites in southern Chile allowed the first identification of a trombiculid species as vector and reservoir of scrub typhus outside the tsutsugamushi triangle, providing unique insights into the ecology and transmission of this recently discovered rickettsial infection in South America., (© The Author(s) 2021. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Scrub typhus is a potentially severe rickettsiosis, caused by Orientia tsutsugamushi in the Asia-Pacific region. Recently, however, two distinct pathogens, " Candidatus Orientia chuto" and " Candidatus Orientia chiloensis", have been discovered in the Middle East and South America, respectively. Since the novel pathogens differ significantly from O. tsutsugamushi , many established diagnostic methods are unreliable. This work describes the development and validation of a new quantitative real-time PCR (qPCR) assay (Orien16S) for the detection of all known Orientia species. Based on a 94 bp sequence of the 16S rRNA gene ( rrs ), Orien16S recognized DNA samples from O. tsutsugamushi ( n = 41), Ca. O. chiloensis ( n = 5), and Ca. O. chuto ( n = 1), but was negative for DNA preparations from closely related rickettsiae and other members of the order Rickettsiales ( n = 22) as well as unrelated bacterial species ( n = 11). After its implementation in Chile, the assay was verified, correctly identifying all tested eschar and buffy coat samples ( n = 28) of clinical suspected cases. Furthermore, Orien16S detected Orientia DNA in trombiculid mites collected in endemic regions in southern Chile. The presented novel qPCR assay provides a useful tool for detecting Orientia and diagnosing scrub typhus from all geographical regions., Competing Interests: JJ and AR are employed by the Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Jiang, Martínez-Valdebenito, Weitzel, Farris, Acosta-Jamett, Abarca and Richards.)

Background: Cystic echinococcosis (CE) caused by Echinococcus granulosus sensu lato (s.l.) is a neglected and underdiagnosed parasitic zoonosis that has a significant socioeconomic impact on rural communities relying on livestock farming. CE is endemic across Latin America, including Chile, where the Coquimbo region exhibits a relatively high record of hospital-based human cases and infected animals. However, the incidence of hospitalized CE cases may underestimate the real burden of infection in a population, since the majority of cases never reach medical attention or official disease records., Methodology/principal Findings: In 2019, a cross-sectional, community-based study was conducted with the objectives of estimating for the first time the prevalence of human abdominal CE using abdominal ultrasound (US) screening in volunteers residing in urban and rural localities of the Monte Patria municipality located in Limarí province, Coquimbo region, Chile, and identifying the risk factors associated with human infection. Pre-screening activities included a 16-h lecture/hands-on training aimed at rural physicians that focused on the diagnosis of CE by US, based on current WHO recommendations. A total of 2,439 (~8% of municipality inhabitants) people from thirteen target localities were screened by abdominal US in June-July 2019. We found an overall CE prevalence of 1.6% (95% CI 1.1-2.2) with a significantly higher likelihood of infection in rural localities, older age classes and people drinking non-potable water; 84.6% of infected volunteers were newly diagnosed with CE. Cysts were either in active or inactive stages in equal proportions; active cysts were detected in all age classes, while 95.7% of inactive cysts occurred in >40 years-old subjects., Conclusions/significance: This is the first US survey aimed at detecting human infection caused by Echinococcus granulosus s.l. in Chile. Our findings indicate a high CE prevalence in the area, and contribute to define the demographic and behavioral risk factors promoting the transmission of the parasitic infection within target communities. Our results support the implementation of cost-effective strategies for the diagnosis, treatment and control of CE, and the need to improve the epidemiological surveillance system in Chile., Competing Interests: The authors have declared that no competing interests exist.

Trichinellosis is a zoonosis with worldwide distribution. Several hosts have been recognized around the world; however, there is a lack of knowledge of the role of feral mammals in Chile in its transmission. Herein, we tested muscle samples from 555 individuals among American minks (Neovison vison Schreber, 1777. n = 100) and several myomorph rodent species (Muridae and Cricetidae. n = 455) from southern Chile by artificial digestion to detect Trichinella larvae and identified the larvae at the species level through molecular analyses. Rodents were captured in agricultural and wild protected areas of several administrative regions (hereafter: region), while minks were captured in agricultural areas of the Los Ríos region. Trichinella spiralis larvae were detected in a synanthropic black rat (0.24%) of the Ñuble region and in seven minks (7%) trapped in agricultural areas of the Los Ríos region. The present results suggest that T. spiralis circulation is restricted to synanthropic rodents and minks living in agricultural areas where the parasite circulates among domesticated pigs. This study represents the first record of T. spiralis in a mustelid mammal in South America, increasing the number of feral species that could participate in the reservoir., (© 2021 Wiley-VCH GmbH.)

Blood samples of 626 rural dogs, 140 Andean foxes ( Lycalopex culpaeus ), and 83 South American grey foxes ( L. griseus ) from six bioregions of Chile spanning 3000 km were screened for Mycoplasma DNA by conventional PCR and sequencing. Risk factors of infection were inferred using Generalized Linear Mixed Models and genetic structure by network analyses. Overall, Mycoplasma haemocanis / Mycoplasma haemofelis ( Mhc / Mhf ) and Candidatus Mycoplasma haematoparvum ( C Mhp) observed prevalence was 23.8% and 12.8% in dogs, 20.1% and 7.2% in Andean foxes, and 26.5% and 8.4% in grey foxes, respectively. Both hemoplasmas were confirmed in all the bioregions, with higher prevalence in those where ticks from the Rhipicephalus sanguineus species group were absent. Candidatus M. haematominutum and a Mycoplasma sp. previously found in South American carnivores were detected in one fox each. Although the most prevalent Mhc/Mhf and C Mhp sequence types were shared between dogs and foxes, network analysis revealed genetic structure of Mhc/Mhf between hosts in some regions. Male sex was associated with a higher risk of Mhc/Mhf and C Mhp infection in dogs, and adult age with C Mhp infection, suggesting that direct transmission is relevant. No risk factor was identified in foxes. Our study provides novel information about canine hemoplasmas with relevance in distribution, transmission routes, and cross-species transmission.

Three species of chigger mites are recorded in our collections from four species of cricetid rodents on Chiloé Island (southern Chile, Los Lagos Region), an area endemic to scrub typhus (Orientia sp.). Two species are described as new-Herpetacarus (Abonnencia) eloisae sp. nov. and Quadraseta chiloensis sp. nov. One species, Paratrombicula goffiStekolnikov and González-Acuña 2012, is for the first time recorded on a mammal host (one species of cricetid rodent), and its distribution is extended to the Los Lagos Region of Chile. The genus ProschoengastiaVercammen-Grandjean, 1967 is synonymized with the subgenus Herpetacarus (Abonnencia)Vercammen-Grandjean, 1960, and four new combinations are established: Herpetacarus (Abonnencia) herniosa (Brennan and Jones, 1961), comb. nov., Herpetacarus (Abonnencia) insolita (Brennan and Jones, 1961), comb. nov., Herpetacarus (Abonnencia) macrochaeta (Brennan and Jones, 1961), comb. nov., and Herpetacarus (Abonnencia) antarctica (Stekolnikov and Gonzalez-Acuña, 2015), comb. nov., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Domestic dogs ( Canis familiaris ) often cohabite at interfaces shared by humans and wildlife, interacting with wild canids as predators, prey, competitors and reservoirs of several multi-host pathogens, such as canid-borne micro and macro parasites that could impact on wildlife, livestock and public health. However, spatio-temporal patterns of indirect interactions as promoters of pathogen transfer between domestic and wild canids are largely unknown. In this study, we used camera traps to describe the activity patterns and habitat use of dogs, chilla ( Lycalopex griseus ) and culpeo ( Lycalopex culpaeus ) foxes and identify the local-scale factors that may affect the frequency of dog-fox interactions through an anthropization gradient of the Coquimbo region, Chile. We assessed local-scale variables that may predict the number of interactions between dogs and foxes, and compared the time interval between dog-culpeo and dog-chilla interactions. Our findings suggested that closeness to urbanized zones predicts the frequency of indirect interactions between dogs and foxes. We found higher number of dog-fox interactions (60 interactions) at a periurban site adjacent to two coastal towns (Tongoy and Guanaqueros), compared to other two more undisturbed sites (12 interactions) increasingly distanced from urbanized areas. We showed that dogs interacted more frequently with chilla foxes (57 interactions) than with culpeo foxes (15 interactions), and the first interaction type occurred almost exclusively at the periurban site, where dogs and chillas were more frequently detected than in the other sites. We detected a marked temporal segregation between dogs and foxes, but dog-chilla interactions resulted in shorter time intervals (2.5 median days) compared to dog-culpeo interactions (7.6 median days), suggesting a higher potential risk of pathogen spillover between the first species pairing. Based on previous studies, we suggest periurban zones may constitute a potential focus of pathogen exposure between dog and fox populations in the study area. Our research contributes to improving the knowledge on the spatio-temporal patterns of interspecific contact between invasive and native carnivores within the context of multi-host pathogen dynamics. Our outcomes will inform theoretical epidemiological models designed to predict and minimize the contact risk between domestic and threatened species, guiding effective control strategies at the wildlife-domestic interface., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Hernández, Manqui, Mejías and Acosta-Jamett.)

In recent years, the spectrum and epidemiology of human rickettsioses has become an emerging topic in Chile. This survey aimed to assess the seroprevalence of spotted fever group rickettsiae (SFGR), typhus group rickettsiae (TGR), and scrub typhus group orientiae (STGO) in northern, central, and southern Chile. We performed a cross-sectional study of healthy adults in rural and urban settings of five regions. Participants were chosen by double stratified random sampling in urban and by convenience in rural locations (n = 1302). Serum specimens were analyzed for group-specific IgG antibodies against SFGR, TGR, and STGO by enzyme-linked immunosorbent assays (ELISAs). Overall seroprevalences to SFGR, TGR, and STGO were 5.3 %, 1.2 %, and 0.4 %, respectively. Prevalences showed geographical differences. Statistical analyses revealed an association of older age with seropositivity to SFGR and to TGR and of rural setting and male gender with seropositivity to SFGR. The study indicates that SFGR, TGR, and STGO are endemic in Chile. The very low STGO seroprevalence might indicate an insufficient sensitivity of serological tests using Asian O. tsutsugamushi strains as ELISA antigens for the detection of antibodies against Chilean Orientia species., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Cystic echinococcosis (CE) is a parasitic zoonosis, caused by the larval stage of Echinococcus granulosus. This disease has a worldwide distribution and is considered an important zoonosis in Chile, especially in regions where raising livestock dominates. We aimed to describe the spatial distribution and risk factors for CE in cattle and sheep slaughtered in the Aysén Region, Chile between 2015 and 2016, to inform disease control in this hyper-endemic region. Clustering and hotspot analyses of CE at the farm-level were performed (Moran's global index, local indicator of spatial autocorrelation (LISA), and the scan statistic Poisson model), and we used spatial interpolation to display areas with a higher risk of CE. Mixed effects logistic regression models were fit to assess the association between municipality of origin and age of animals and CE detected at slaughter, with the farm of origin included as a random effect. Overall, data from 1532 cattle farms (1078 geocoded) and 30,805 cattle were analyzed, with a 64 % and 39 % CE prevalence at the farm and animal-level, respectively. For sheep farms, data from 381 (296 geocoded) farms and 58,223 sheep were analyzed, with a 71 % and 18 % CE prevalence at the farm and animal-level, respectively. Globally, the CE prevalence in both cattle and sheep farms was not clustered. However, we found 14 cattle and 23 sheep farms with high prevalence that were locally clustered. We also detected 6 and 11 clusters in cattle and sheep farms, respectively. Risk of reported CE varied according to municipalities of origin, and the age of animals. This study confirms that the Aysén region is a hyper-endemic region for CE. It demonstrates the usefulness of the traceability of animals slaughtered for understanding the epidemiology of CE. The results suggest that in this hyper-endemic region in which there are limited financial resources available, that intervention activities - at the beginning of a control program - should be focused on farms with high prevalence and identified disease clusters., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Scrub typhus is a potentially fatal rickettsiosis caused by Orientia species intracellular bacteria of the genus Orientia. Although considered to be restricted to the Asia Pacific region, scrub typhus has recently been discovered in southern Chile. We analyzed Orientia gene sequences of 16S rRNA (rrs) and 47-kDa (htrA) from 18 scrub typhus patients from Chile. Sequences were ≥99.7% identical among the samples for both amplified genes. Their diversity was 3.1%-3.5% for rrs and 11.2%-11.8% for htrA compared with O. tsusugamushi and 3.0% for rrs and 14.8% for htrA compared with Candidatus Orientia chuto. Phylogenetic analyses of both genes grouped the specimens from Chile in a different clade from other Orientia species. Our results indicate that Orientia isolates from Chile constitute a novel species, which, until they are cultivated and fully characterized, we propose to designate as Candidatus Orientia chiloensis, after the Chiloé Archipelago where the pathogen was identified.

Background: Pet-related tick-borne infections constitute an emerging problem in human and veterinary medicine worldwide. In Chile, two pathogens of the Anaplasmataceae family, Anaplasma platys and Ehrlichia canis , have been identified in recent years in dogs and vectors. This epidemiological survey aimed to determine the human and canine seroprevalence of Anaplasma spp. in urban and rural areas of different regions in Chile and to analyze the factors associated with seropositivity. Methods: We performed a cross-sectional household-based study in four regions, where healthy adults and their household dogs were included. Households were chosen by double stratified random sampling in urban areas and by convenience sampling in rural areas. Anaplasma seroreactivity was determined by a commercial microimmunofluorescence assay using Anaplasma phagocytophilum antigens. A questionnaire survey was applied to assess risk factors for Anaplasma seropositivity. Results: A total of 1105 persons and 905 dogs were included. The overall seroprevalence in humans was 9.4%, ranging from 5.6% in central Chile to 13.7% in the extreme north; in dogs the overall prevalence was 44.0% and ranged from 37.2% in the northern region to 61.1% in central Chile. Risk factors for human seropositivity were lower education and external deworming of dogs, whereas canine seropositivity was associated to urban site, mixed-breed, older animals, and tick infestation. Conclusions: This cross-sectional study suggests a broad exposure of both dogs and dog owners to Anaplasma or related agents in Chile. Further epidemiological and clinical studies are necessary to understand the complete spectrum and epidemiology of tick-borne zoonotic infections in the different ecoclimatic regions of Chile.

Background: Scrub typhus is an emerging vector-borne zoonosis, caused by Orientia spp. and transmitted by larvae of trombiculid mites, called chiggers. It mainly occurs within a region of the Asia-Pacific called the tsutsugamushi triangle, where rodents are known as the most relevant hosts for the trombiculid vector. However, the reservoir(s) and vector(s) of the scrub typhus outside Asia-Pacific are unknown. The disease has recently been discovered on and is considered endemic for Chiloé Island in southern Chile. The aim of the present work was to detect and determine the prevalence of chiggers on different rodent species captured in probable sites for the transmission of orientiae responsible for scrub typhus on Chiloé Island in southern Chile and to molecularly examine collected chiggers for the presence of Orientia DNA., Methodology/principal Findings: During the austral summer 2018, rodents were live-trapped in six sites and examined for chigger infestation. All study sites were rural areas on Chiloé Island, previously identified as probable localities where human cases acquired the scrub typhus. During a total of 4,713 trap-nights, 244 rodents of seven species were captured: the most abundant was Abrothrix olivacea. Chiggers were detected on all seven rodent species with a 55% prevalence rate. Chiggers showed low host specificity and varied according to site specific host abundance. Three genera of trombiculids were identified. Herpetacarus was the most abundant genus (93%), prevalent in five of the six sites. Infestation rates showed site specific differences, which were statistically significant using a GLM model with binomial errors. Molecular analyses proved that 21 of 133 (15.8%) mite pools were positive for Orientia species, all of them belonged to the genus Herpetacarus., Conclusions/significance: This study firstly reports the presence of different rodent-associated chigger mites positive for Orientia sp., in a region endemic for scrub typhus in southern Chile. Herpetacarus and two other genera of mites were found with high infestation rates of rodents in sites previously identified as probable exposure of scrub typhus cases. A substantial percentage of mite pools were positive for Orientia DNA, suggesting that chigger mites serve as vectors and reservoirs of this emerging zoonosis in South America., Competing Interests: The authors have declared no competing interests.

Cystic echinococcosis (CE) is a zoonotic parasitic disease associated with Echinococcus granulosus . The parasite is maintained by domestic and wild canids as definitive hosts with several ungulate species as intermediate hosts in domestic and peridomestic transmission cycles. In Chile, CE is endemic, and the role of livestock and dogs ( Canis lupus familiaris ) in the cycle and the accidental infection of humans are widely documented at rural sites. However, the role of wild herbivores in wild cycles or the potential transmission of CE from livestock is still unknown in Chile and the rest of South America. We used molecular techniques to describe CE infecting a Patagonian huemul ( Hippocamelus bisulcus ) in Cerro Castillo National Reserve (Aysén region, Chile). We make inferences about the risk of disease spillover from sympatric domestic and wild species. The DNA-based molecular analysis revealed that the huemul was infected with E. granulosus G1 genotype, sharing haplotypes with other G1 samples collected from sheep ( Ovis aries ) and cattle ( Bos taurus ) worldwide. Geographic overlap between sheep and huemul populations in the reserve likely facilitates parasite spillover into wild deer populations, with shepherd or stray dogs and wild foxes ( Lycalopex culpaeus ) potentially acting as bridging hosts between livestock and the endangered huemul. Further studies are warranted to understand the implications of E. granulosus for huemul conservation throughout the Chilean Patagonia.

Endemic scrub typhus was recently detected on Chiloé Island in southern Chile. We report a series of cases, acquired over a wide geographical range in continental Chile during 2016-2018, demonstrating that this emerging rickettsial infection is also found on the mainland of South America.

Marketing is a stressful process for beef calves, because they are removed from their environment, often weaned just before loading, loaded, transported, and unloaded. It also involves extended periods without food and water and mixing with unfamiliar animals in an unknown environment. Some studies have shown that calves sold through markets are exposed to extended fasting periods even when they undergo only short transportation times. The aim of this controlled study was to determine the consequences for beef calves of a short transportation time followed by a prolonged time without food and water on their tympanic temperature (TT), maximum eye temperature (MET), blood variables related to stress, and live weight. Ten calves were transported for 3 h and then kept in an outdoor pen for 21 h, completing a 24 h fasting period. Sampling took place before loading, after transport and unloading, and then after completing 24 h without food and water. TT, MET, blood glucose, and creatine kinase (CK) increased significantly after transportation. Live weight decreased across sample times (mean of 10 kg per calf after 24 h of fasting), which was consistent with the higher concentration of β-HB found after fasting. Further studies to measure the actual consequences of true, commercial marketing on calf welfare and productivity are needed., Competing Interests: The authors declare no conflicts of interest.

Cystic echinococcosis (CE), a zoonotic disease caused by tapeworms of the species complex Echinococcus granulosus sensu lato, represents a substantial global health and economic burden. Within this complex, E. granulosus sensu stricto (genotypes G1 and G3) is the most frequent causative agent of human CE. Currently, there is no fully reliable method for assigning samples to genotypes G1 and G3, as the commonly used mitochondrial cox1 and nad1 genes are not sufficiently consistent for the identification and differentiation of these genotypes. Thus, a new genetic assay is required for the accurate assignment of G1 and G3. Here we use a large dataset of near-complete mtDNA sequences (n = 303) to reveal the extent of genetic variation of G1 and G3 on a broad geographical scale and to identify reliable informative positions for G1 and G3. Based on extensive sampling and sequencing data, we developed a new method, that is simple and cost-effective, to designate samples to genotypes G1 and G3. We found that the nad5 is the best gene in mtDNA to differentiate between G1 and G3, and developed new primers for the analysis. Our results also highlight problems related to the commonly used cox1 and nad1. To guarantee consistent identification of G1 and G3, we suggest using the sequencing of the nad5 gene region (680 bp). This region contains six informative positions within a relatively short fragment of the mtDNA, allowing the differentiation of G1 and G3 with confidence. Our method offers clear advantages over the previous ones, providing a significantly more consistent means to distinguish G1 and G3 than the commonly used cox1 and nad1., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Echinococcus granulosus sensu stricto (s.s.) is the major cause of human cystic echinococcosis worldwide and is listed among the most severe parasitic diseases of humans. To date, numerous studies have investigated the genetic diversity and population structure of E. granulosus s.s. in various geographic regions. However, there has been no global study. Recently, using mitochondrial DNA, it was shown that E. granulosus s.s. G1 and G3 are distinct genotypes, but a larger dataset is required to confirm the distinction of these genotypes. The objectives of this study were to: (i) investigate the distinction of genotypes G1 and G3 using a large global dataset; and (ii) analyse the genetic diversity and phylogeography of genotype G1 on a global scale using near-complete mitogenome sequences. For this study, 222 globally distributed E. granulosus s.s. samples were used, of which 212 belonged to genotype G1 and 10 to G3. Using a total sequence length of 11,682 bp, we inferred phylogenetic networks for three datasets: E. granulosus s.s. (n = 222), G1 (n = 212) and human G1 samples (n = 41). In addition, the Bayesian phylogenetic and phylogeographic analyses were performed. The latter yielded several strongly supported diffusion routes of genotype G1 originating from Turkey, Tunisia and Argentina. We conclude that: (i) using a considerably larger dataset than employed previously, E. granulosus s.s. G1 and G3 are indeed distinct mitochondrial genotypes; (ii) the genetic diversity of E. granulosus s.s. G1 is high globally, with lower values in South America; and (iii) the complex phylogeographic patterns emerging from the phylogenetic and geographic analyses suggest that the current distribution of genotype G1 has been shaped by intensive animal trade., (Copyright © 2018 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Background: Scrub typhus is a potentially life-threatening vector-borne infection caused by Orientia species. It occurs mainly in the Asian-Pacific region, where it causes significant morbidity and mortality. Recently, an endemic focus of scrub typhus has been described in South America, on Chiloé Island in southern Chile. Dogs have been used as sentinel hosts to determine the presence and spatial distribution of various vector-borne infections. Their suitability to gain insight into human exposure to Orientia tsutsugamushi has been suggested in studies from Asia., Methodology: In January 2016, we conducted a cross-sectional study, which included the two main cities on Chiloé Island. Canine blood samples were obtained in households, chosen by double stratified random sampling in urban and by convenience in rural locations. Specimens were tested by ELISA for IgG antibodies against whole-cell antigen preparations from three strains of O. tsutsugamushi. Data were further analyzed for factors associated with seropositivity including spatial clustering., Results: Serum samples from 202 dogs (104 urban, 98 rural) were tested for IgG against O. tsutsugamushi, of which 43 (21.3%) were positive. Seroprevalence rates were higher in rural than in urban settings (p<0.01) and in older compared to younger dogs (p<0.01). Spatial analysis by LISA indicated the presence of four localities of highly grouped cases., Conclusions: The detected seroprevalence supports the endemicity of scrub typhus in southern Chile and suggests a wide exposure of household dogs to the infected, yet unknown vector(s). The spatial data will be used for future research identifying further human cases as well as the local vector(s)/reservoirs for scrub typhus in southern Chile. The study reinforces that dogs are useful sentinels for Orientia spp. in regions of uncertain endemicity and distribution., Competing Interests: The authors have declared that no competing interests exist.

Scrub typhus, caused by Orientia tsutsugamushi and transmitted by larvae of trombiculid mites, is an endemic rickettsiosis in the Asia Pacific region. After the first identification of a case in Chile in 2006, more than 30 cases have been diagnosed by our group since 2015. Cases were detected predominantly during the Chilean summer months. Patients presented with fever, rash, and a typical eschar at the inoculation site; other frequent findings were intense headache, night sweats, increased laboratory markers of inflammation and transaminases. The vast majority of cases have been diagnosed in southern Chile (mainly Chiloé Island), although recently some cases were also identified in the central Metropolitan Region in patients returning from trips to southern Chile. Physicians attending Chilean patients should be aware of this emerging infection to be able to initiate empirical therapy with doxycycline. The confirmation of cases by the diagnostic methods available in Chile will contribute to a better understanding of the epidemiological and clinical relevance of this emerging infection in South America.

Cystic echinococcosis, a zoonotic disease caused by Echinococcus granulosus sensu lato (s. l.), is a significant global public health concern. Echinococcus granulosus s. l. is currently divided into numerous genotypes (G1-G8 and G10) of which G1-G3 are the most frequently implicated genotypes in human infections. Although it has been suggested that G1-G3 could be regarded as a distinct species E. granulosus sensu stricto (s. s.), the evidence to support this is inconclusive. Most importantly, data from nuclear DNA that provide means to investigate the exchange of genetic material between G1-G3 is lacking as none of the published nuclear DNA studies have explicitly included G2 or G3. Moreover, the commonly used relatively short mtDNA sequences, including the complete cox1 gene, have not allowed unequivocal differentiation of genotypes G1-G3. Therefore, significantly longer mtDNA sequences are required to distinguish these genotypes with confidence. The main aim of this study was to evaluate the phylogenetic relations and taxonomy of genotypes G1-G3 using sequences of nearly complete mitogenomes (11,443bp) and three nuclear loci (2984bp). A total of 23 G1-G3 samples were analysed, originating from 5 intermediate host species in 10 countries. The mtDNA data demonstrate that genotypes G1 and G3 are distinct mitochondrial genotypes (separated by 37 mutations), whereas G2 is not a separate genotype or even a monophyletic cluster, but belongs to G3. Nuclear data revealed no genetic separation of G1 and G3, suggesting that these genotypes form a single species due to ongoing gene flow. We conclude that: (a) in the taxonomic sense, genotypes G1 and G3 can be treated as a single species E. granulosus s. s.; (b) genotypes G1 and G3 should be regarded as distinct genotypes only in the context of mitochondrial data; (c) we recommend excluding G2 from the genotype list., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is a new and revolutionary identification method for microorganisms and has recently been introduced into clinical microbiology in many industrialized countries in Europe and North America., Objectives: Our study aimed to compare the performance and practicality of two commercial MALDI-TOF MS platforms in a head-to head manner at a routine laboratory in Chile., Methods: During a five-month period in 2012-13, the diagnostic efficiency (correct identification rate) and agreement between Microflex LT (Bruker Daltonics) and Vitek MS (bioMérieux) was compared in a parallel manner to conventional identification including genotypic analysis for difficult-to-identify strains. The study included 804 microbial isolates: 252 Enterobacteriaceae, 126 non-fermenters, 36 other gram-negative rods, 279 gram-positive cocci, 32 gram-positive rods, 32 anaerobes, and 47 yeasts. Other relevant factors of the two devices such as user friendliness and connectivity were also evaluated and compared., Results: Both systems correctly identified the vast majority (98%) of the isolates to the genus level. Vitek MS reached higher rates of identification to species and species complex level than Microflex LT (81% vs. 85% and 87% vs. 93%, respectively), which was mainly based on the higher performance among coagulase negative staphylococci and Candida isolates. The evaluation of user friendliness and other technical aspects showed only marginal differences, which slightly favored Vitek MS, mainly due to its ready-to-use supplies, easier connectivity and workflow integration, and availability of local technical support., Conclusions: Both MALDI-TOF MS systems permitted fast and accurate identification of most microbial strains and showed a high level of user-friendliness. The observed differences were marginal and slightly favored Vitek MS, mainly due to practicality and connectivity issues within our setting.