Pathological lesions associated with avian schistosomes (Digenea: Schistosomatidae) in the freshwater snail Chilina dombeiana (Gastropoda: Chilinidae) from Southern Chile.

Avian schistosomes inhabit the blood stream of domestic and wild birds with aquatic snails as their intermediate hosts. In the Neotropics there is an emerging effort to describe species from these hosts, including Chile, although the knowledge about their pathological consequences is mostly understudied. This study aimed to describe the pathological changes associated with the parasitism of a native schistosomatid restricted to the Southern Cone of Neotropics. To achieve this, a total of 401 Chilina dombeiana snails (Chilinidae) were collected in two locations from Southern Chile. All of them were disposed to cercarial release procedure for three consecutive days. Furcocercariae released were stained and characterized by microscopic evaluation. Then, all snails were dissected under stereomicroscope and preserved in 10 % buffered formalin until histopathological analysis was performed. Eight out 401 (P = 2 %) snails were found parasitized with avian schistosomes. The released furcocercariae were identified as Schistosomatidae gen. sp. Lineage II which was previously reported in the same host. The main pathological change was an atrophy of ovotestes and an absence or mild infiltration of hemocytes in the surrounding tissues. Besides, a co-infection with echinostomes was found which was associated with a moderate hemocyte infiltration, granuloma-like lesion, and a reduced presence of schistosome' sporocysts. The latter would suggest an antagonistic interaction between these two digeneans, as has been proposed in the Echinostoma spp.-Schistosoma mansoni model. Despite the above, the release of furcocercariae was present but reduced, in contrast with the non-release of echinocercariae. This interaction requires further attention. This study represents the first attempt to characterize the pathological consequences of parasitism by a native, yet undescribed, avian schistosome in an endemic snail. Future studies should consider experimental infections to understand the dynamics of single infections in other Chilina species, including inter- and intra-specific parasitism as previous studies have found, including this study.

Phylogenetic and ecological factors affecting the sharing of helminths between native and introduced rodents in Central Chile.

In order to analyse the effect of hosts' relationships and the helminthic load on the switching of parasites between native and introduced hosts, we sampled rodents belonging to two suborders from Central Chile. We compared the number of helminthic species shared between murids (introduced) and cricetid (native, same suborder) rodents to those shared between murids and hystricomorphs (native, different suborder), and we assessed the association between parasitic presence, abundance and geographical dispersion in source hosts to the presence and abundance in recipient hosts. Introduced rodent species shared more helminth species with cricetid rodents than with non-cricetids. Presence and abundance in recipient hosts was not associated with the prevalence and mean abundance in source hosts' population. The mean abundance of parasites in source hosts throughout the territory and wider dispersion was positively associated with the likelihood of being shared with a recipient host. Closer relationships between native and introduced hosts and high parasitic abundance and dispersion could facilitate host switching of helminths between native and introduced rodents. This work provides the first documentation of the importance of parasitic abundance and dispersion on the switching of parasites between native and introduced hosts.

[Analysis of human cyst echinococcosis in a northern Chilean region from 2008 to 2012]. / Análisis de la hidatidosis humana en la Región de Coquimbo entre los años 2008 y 2012.

BACKGROUND: The Coquimbo Region in Chile has the highest incidence of hydatidosis in central and northern Chile. AIM: To analyze the incidence of human cystic echinococcosis (CE) in the Coquimbo Region between 2008 and 2012. MATERIAL AND METHODS: Official records of reported cases were analyzed. The association of the location, age and sex with the presence/absence of CE was analyzed by means of logistic regressions. The incidence was reported as cases per 100,000 inhabitants. RESULTS: The mean annual incidence was 2.6 cases per 100,000 inhabitants, which ranged from 3.1 (2010) to 1.9 (2012) cases. The districts with higher incidence were Punitaqui (21.2 cases), Paihuano (9 cases), Rio Hurtado (8 cases), Canela (8.1 cases), Monte Patria (7.9 cases), Vicuña (6.9 cases) and Combarbalá (6.9 cases). The incidence in these locations was significantly higher than elsewhere in the region. Males had a significantly higher incidence than females (3.3 and 1.8 respectively). Incidence increased significantly with age, notably increasing after 45 years of age. CONCLUSIONS: Rural locations had the highest incidences of CE in the Coquimbo Region. The disease was reported more commonly in adults and mainly in men.

Molecular survey of tick-borne infectious agents in cricetid rodents (Rodentia: Cricetidae) in Central and Southern Chile.

Tick-borne infectious agents (TBIAs) include several bacteria and protozoa that can infect vertebrates, including humans. Some of these agents can cause important diseases from both a public health perspective, such as Lyme disease, and from an animal health and production viewpoint, such as Texas fever. In Chile, several studies have assessed the presence of tick-borne disease agents in vectors and mammal hosts, mainly in the northern regions, but few studies have assessed the presence of these agents in Central and Southern Chile. This study aimed to assess the presence of three groups of TBIAs-Borrelia, Anaplasmataceae, and Piroplasmida-in cricetid rodents of Central and Southern Chile. A total of 207 specimens from 13 localities between the O'Higgins and Los Lagos regions were captured. DNA was extracted from the liver and spleen, and subsequently underwent polymerase chain reaction (PCR) amplification targeting the 16S rRNA, flaB, and 18S rRNA genes to detect DNA from Borrelia, Anaplasmataceae, and Piroplasmida, respectively. Although no DNA from these TBIAs was detected, the DNA extraction process was validated by optimal DNA purity ratios (an A260/A280 ratio within the 1.6-2.0 range) and successful internal control amplification in all samples. These results, in addition to findings from previous reports, suggest a very low prevalence of these TBIAs in the rodent population studied. Further research into the factors influencing the presence of these agents and their vectors will provide insight into the reasons underlying this low prevalence.

Gastrointestinal parasites in wild rodents in Chiloé Island-Chile.

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.

Survey and Molecular Characterization of Sarcocystidae protozoa in Wild Cricetid Rodents from Central and Southern Chile.

In Chile, studies of parasites from the family Sarcocystidae (Apicomplexa) have mostly been related to domestic animals. We aimed to assess the presence of Sarcocystidae taxa in cricetid rodents from Central and Southern Chile. We studied 207 rodents, encompassing six species, from 13 localities. We isolated DNA from tissue samples, amplified the Sarcocystidae 18S rRNA gene with polymerase chain reaction, and performed phylogenetic analyses using maximum likelihood and Bayesian inferences. In addition, we examined blood smears and performed histological studies in organs from Sarcocystidae DNA-positive animals. Three specimens were DNA-positive and three genotypes were retrieved and named: Sarcocystis sp. P61, related to Sarcocystis strixi, was detected in two Abrothrix olivacea. Toxoplasmatinae gen. sp. P99 was retrieved from those same two specimens, and was related to Toxoplasma and other genera, although it branched independently. Besnoitia sp. R34 was detected in one Abrothrix hirta, and was clustered with congeneric species associated with rodents. No protozoa were found during microscopic studies; thus, it was not possible to confirm parasitic interactions rather than accidental encounters. However, the close relatedness of the retrieved genotypes to parasites of rodents supports the hypothesis of host-parasite associations. All three genotypes are suggested as potential new taxa, including a putative new genus.

Molecular surveillance of potential SARS-CoV-2 reservoir hosts in wildlife rehabilitation centers.

BACKGROUND: The COVID-19 pandemic, caused by SARS-CoV-2 infection, has become the most devastating zoonotic event in recent times, with negative impacts on both human and animal welfare as well as on the global economy. Although SARS-CoV-2 is considered a human virus, it likely emerged from animals, and it can infect both domestic and wild animals. This constitutes a risk for human and animal health including wildlife with evidence of SARS-CoV-2 horizontal transmission back and forth between humans and wild animals. AIM: Molecular surveillance in different wildlife rehabilitation centers and wildlife associated institutions in Chile, which are critical points of animal-human interaction and wildlife conservation, especially since the aim of wildlife rehabilitation centers is to reintroduce animals to their original habitat. MATERIALS AND METHODS: The survey was conducted in six WRCs and three wildlife associated institutions. A total of 185 samples were obtained from 83 individuals belonging to 15 different species, including vulnerable and endangered species. Each specimen was sampled with two different swabs: one oropharyngeal or nasopharyngeal according to the nostril diameter, and/or a second rectal sample. RNA was extracted from the samples and two different molecular assays were performed: first, a conventional RT-PCR with pan-coronavirus primers and a second SARS-CoV-2 qPCR targeting the N and S genes. RESULTS: All 185 samples were negative for SARS-CoV-2. CLINICAL RELEVANCE: This study constitutes the first report on the surveillance of SARS-CoV-2 from wildlife treated in rehabilitation centers in Chile, and supports the biosafety procedures adopted in those centers.

The genus Trichinella and its presence in wildlife worldwide: A review.

The genus Trichinella has a worldwide distribution, infecting people, domestic animals, and wildlife. It includes 13 genotypes, which are geographically delimited; Trichinella is transmitted to people through the ingestion of undercooked meat. Historically, it has been associated with pigs, but most Trichinella species affect wildlife, and cases of trichinellosis due to the consumption of game meat have been emerging. Therefore, it is important to monitor the sources of transmission to domestic animals and humans. The objective of this work was to analyse reports of Trichinella spp. in wild/feral animals around the world to identify the needs of future research in the epidemiology of the sylvatic cycle. A search of studies published until 2021 was conducted using Web of Science and SciELO. In the Palearctic, the most commonly studied hosts were wild boars and red foxes, and hosts with the highest prevalence rates were polar bears and martens. In the Nearctic, red foxes and black bears were the most frequently studied hosts, and the highest prevalence was found for wolverines and brown bears. In the Neotropics, positive reports were only identified in two countries, with wild boars being the most commonly studied species, and armadillos featuring the highest prevalence. In the Afrotropics, Trichinella limits its presence to Sub-Saharan Africa, where lions are the most studied hosts, and spotted hyenas have the highest prevalence. In the Indo-Malaya and Australasia ecozones, information on wildlife is scarce; the Norwegian rat is the most frequently studied host, and the Tasmanian devil has the highest prevalence of infection. In the last decade, research on world wildlife has increased which is associated with more frequent trichinellosis outbreaks caused by the consumption of wild meat. The results suggest the need to increase research in developing countries, particularly where more diverse sources of meat are available for human consumption.

Assessment of the Vaccination Program against Cystic Echinococcosis in Sheep in the Pehuenche Community of Central Chile.

Echinococcosis is a neglected zoonosis that uses dogs and sheep as its main hosts in Chile. The Eg95 vaccine against sheep infection has been included in some control programs. Here, we assess the efficacy of the vaccination program in the hyperendemic Alto Biobío commune after 3 years of execution. Fisher's test and generalized linear models were used in the assessment. The program tried to offer a first dose at 2 months of age, a booster 1 month later, and yearly vaccination. Given logistic difficulties, important delays in vaccination occurred, and most animals did not receive the first booster. Dog deworming was not included in the program. Likely due to the aforementioned factors, the overall frequency of infection was not lower, but the proportion of large (>5 mm) cysts and fertile cysts was smaller after the program. The frequency of infection and/or the number of cysts were lower when the age at first dose was younger and the first booster was administered 1 month after the first dose. The results suggest that vaccination affects both cyst development after the larvae reach the target organs, as well as the development of the protoscolex once the cysts start developing.

Patterns of Gastrointestinal Helminth Infections in Rattus rattus, Rattus norvegicus, and Mus musculus in Chile.

Few studies have assessed the patterns of parasite populations of rodents over a longitudinal gradient in Chile. In this work, the gastrointestinal helminthic fauna of invasive rodents in Chile was examined to assess the association between their presence/absence and abundance with latitude, host sex, and host body condition, and to assess the coexistence and correlation of the abundance between parasite species. Rodents were obtained from 20 localities between 33 and 43°S. Helminths were extracted from the gastrointestinal tract and identified morphologically. Overall, 13 helminth taxa were obtained. The most frequently identified parasite species was Heterakis spumosa, and the most abundant was Syphacia muris, while Physaloptera sp. was the most widely distributed. No locality presented with a coexistence that was different from that expected by chance, while the abundance of five helminthic species correlated with the abundance of another in at least one locality, most likely due to co-infection rather than interaction. Host sex was associated with parasite presence or abundance, and female sex-biased parasitism was notably observed in all cases. Body condition and latitude presented either a positive or negative association with the presence or abundance of parasites depending on the species. It is notable that the likely native Physaloptera sp. is widely distributed among invasive rodents. Further, gravid females were found, suggesting spillback of this species to the native fauna. The low frequency and abundance of highly zoonotic hymenolepid species suggest that rodents are of low concern regarding gastrointestinal zoonotic helminths.

Temporal and geographic analysis of trichinellosis incidence in Chile with risk assessment.

Trichinellosis is a foodborne disease caused by several Trichinella species around the world. In Chile, the domestic cycle was fairly well-studied in previous decades, but has been neglected in recent years. The aims of this study were to analyze, geographically, the incidence of trichinellosis in Chile to assess the relative risk and to analyze the incidence rate fluctuation in the last decades. Using temporal data spanning 1964-2019, as well as geographical data from 2010 to 2019, the time series of cases was analyzed with ARIMA models to explore trends and periodicity. The Dickey-Fuller test was used to study trends, and the Portmanteau test was used to study white noise in the model residuals. The Besag-York-Mollie (BYM) model was used to create Bayesian maps of the level of risk relative to that expected by the overall population. The association of the relative risk with the number of farmed swine was assessed with Spearman's correlation. The number of annual cases varied between 5 and 220 (mean: 65.13); the annual rate of reported cases varied between 0.03 and 1.9 cases per 105 inhabitants (mean: 0.53). The cases of trichinellosis in Chile showed a downward trend that has become more evident since the 1980s. No periodicities were detected via the autocorrelation function. Communes (the smallest geographical administrative subdivision) with high incidence rates and high relative risk were mostly observed in the Araucanía region. The relative risk of the commune was significantly associated with the number of farmed pigs and boar (Sus scrofa Linnaeus, 1758). The results allowed us to state that trichinellosis is not a (re)emerging disease in Chile, but the severe economic poverty rate of the Mapuche Indigenous peoples and the high number of backyard and free-ranging pigs seem to be associated with the high risk of trichinellosis in the Araucanía region.

Intestinal Helminths in Wild Rodents from Native Forest and Exotic Pine Plantations (Pinus radiata) in Central Chile.

Native forests have been replaced by forestry plantations worldwide, impacting biodiversity. However, the effect of this anthropogenic land-use change on parasitism is poorly understood. One of the most important land-use change in Chile is the replacement of native forests by Monterey pine (Pinus radiata) plantations. In this study, we analyzed the parasitism (presence and prevalence) of intestinal helminths from fecal samples of wild rodents in three habitat types: native forests and adult and young pine plantations in central Chile. Small mammals were sampled seasonally for two years, and a total of 1091 fecal samples from seven small mammal species were analyzed using coprological analysis. We found several helminth families and genera, some of them potentially zoonotic. In addition, new rodent-parasite associations were reported for the first time. The overall helminth prevalence was 16.95%, and an effect of habitat type on prevalence was not observed. Other factors were more relevant for prevalence such rodent species for Hymenolepis sp. and season for Physaloptera sp. Our findings indicate that pine plantations do not increase helminth prevalence in rodents compared to native forests.

Molecular Survey of Parvoviruses and Mycoplasma spp. in Invasive American Mink (Neovison vison) from Southern Chile.

Using PCR, we evaluated the presence of parvoviruses and Mycoplasma spp. in 123 American mink (Neovison vison), an introduced invasive carnivore in Chile. Our results showed all analyzed animals were negative for both pathogen groups. We cannot completely dismiss their presence, but if present, their prevalence should be lower than 2%.

First record of Trichinella in Leopardus guigna (Carnivora, Felidae) and Galictis cuja (Carnivora, Mustelidae): new hosts in Chile.

BACKGROUND: Trichinellosis is a zoonotic disease with a worldwide distribution. It is caused by several species of nematodes in the genus Trichinella. Trichinella spp. are transmitted through predation or carrion consumption and occur in domestic and sylvatic cycles. In humans trichinellosis occurs due to the consumption of raw or undercooked, infected meat and is mainly associated with the household slaughter of pigs or the consumption of game animals without veterinary inspection, a cultural practice that is difficult to resolve. Therefore, knowledge of this parasite's reservoir is relevant for better implementing public health strategies. The aim of this study was to assess the presence of Trichinella sp. in several carnivore and omnivore vertebrates in central-southern Chile. METHODS: We collected muscle tissue from a total of 53 animals from 15 species and were digested to detect Trichinella larvae which were further identified to species level using molecular techniques. RESULTS: We detected Trichinella larvae in Leopardus guigna (Felidae) and Galictis cuja (Mustelidae). We identified the larvae collected from L. guigna as Trichinella spiralis, but we were unable to molecularly characterize the larvae from G. cuja. This is the first record of Trichinella in a native mustelid of South America and the first record of T. spiralis in L. guigna. This study identified two novel hosts; however, further work is needed to identify the role that these and other hosts play in the cycle of Trichinella in Chile.

Parasites of Native and Invasive Rodents in Chile: Ecological and Human Health Needs.

Invasive populations are a threat to biodiversity, resulting in the loss of species, and also a threat to human health, participating in the reservoir of diseases. Rodents are among the most important invasive species worldwide. Chile is a country that features island conditions in terms of geography and has been widely invaded by allochthonous rodents. In this mini-review, we updated the literature on macro-parasites infecting both native and invasive rodents and of vector-borne pathogens in continental Chile in order to assess the relative importance of invasive rodents from both ecological and public health points of view. A total of 174 parasite species were found, with Siphonaptera representing the most diverse group. When examining how parasites are shared between native and introduced rodents, the analysis suggests that parasites circulate freely within recipient populations, and are not significantly transmitted from source populations. Further, generalist parasites are typically more prone to being shared between native and introduced rodents. Most zoonotic parasites were reported in invasive rodents, suggesting that these rodents must represent a public health concern. Although several vector-borne pathogens have been reported in rodents or ectoparasites, most of the recently emerging research has illustrated that there is a lack of evidence on rodent-vector-borne zoonoses in most pathogens.

Trichinella spiralis in a cougar (Puma concolor) hunted by poachers in Chile.

Trichinellosis is a zoonosis caused by nematodes of the Trichinella genus, of which 10 species have been described. Species identification when larvae is found is only possible using molecular tools. Meat from pigs and game animals not subjected to veterinary inspection are the main sources of human infections. The hunting of native carnivores is prohibited in Chile due to conservation issues and the fact that those animals favor pest control. The illegal hunting of a cougar (Puma concolor) occurred in September 2020. Herein, the molecular identification of Trichinella larvae, by analyzing nuclear (expansion segment V) and mitochondrial (cytochrome C oxidase subunit I) sequences are described. Both the amplification of the expansion segment V region and the phylogenetic analysis of a segment of a fragment of the cytochrome c-oxidase subunit I sequence confirmed that the larvae belonged to T. spiralis. The case described herein represents the first evidence of illegal hunting of a protected mammal infected with Trichinella in Chile, highlighting the 'One Health' perspective to face this disease in the rural-sylvatic interphase.

Survey of Trichinella in American minks (Neovison vison Schreber, 1777) and wild rodents (Muridae and Cricetidae) in Chile.

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.

Factors associated with the presence of helminth eggs in the soil of public areas in Concepción, Chile.

The aim of this study was to assess the association between the distance to the nearest feces, the type of substrate to be studied (feces, soil, grass), and whether a given area was licensed or not with the presence of helminth eggs in Concepción, Chile. A total of 256 samples taken from feces and either from soil or grass at 10 cm, 50 cm, and 100 cm from feces (including 16 public areas), were examined for the presence of parasites using the ZnSO4 technique. The association between variables with the presence of eggs was assessed with multifactorial logistic regressions. 24.21% of samples presented at least one egg. The higher odds of Toxocara egg presence was positively associated with the licensed 'area type' and the soil 'sample type' (when compared with feces). The odds of Ancylostomatidae egg presence was positively associated with the licensed 'area type', but negatively associated with the soil 'sample type' (compared with feces). The results suggest that finding eggs must be interpreted differently based on the parasite species and substrate to be analyzed, and that the presence of parasitized dogs is a more important contributing factor than the frequency with which the ground is cleaned of feces.

Flukicide efficacy against Fasciola hepatica of Triclabendazole and Nitroxynil in cattle of the central valley of Chile.

On a farm with permanent history of fasciolasis a study was performed aimed to know the efficacy of triclabendazole (TCBZ) and then to contrast with that of nitroxynil. Thirty-nine cattle naturally infected with Fasciola hepatica were randomly allocated into 4 experimental groups: Group 1 (control) was left untreated. Group 2 was treated with of 12 mg/kg body weight (bw) of TCBZ by oral route. Group 3 treated with 24 mg/kg bw TCBZ orally. Group 4 was treated with 10 mg/kg bw of nitroxynil subcutaneously. The anthelmintic efficacy was calculated as the percentage of reduction in faecal egg count (FEC) at 14 and 28 d post-treatment. Results indicated that there were no significant differences in the percentage of FEC reduction between control group and the groups treated with 12 or 24 mg/kg of TCBZ. On the contrary, the treatment with nitroxinyl significantly reduced the FEC and decreased the percentage of positive animals. In conclusion, Fasciola hepatica is reported for first time as resistant to TCBZ in Chile, which highlights the need of rotating drugs and assessing the efficacy of the administered drug in order to avoid the selection of resistant worms.

Helminthic infection in three native rodent species from a semiarid Mediterranean ecosystem.

Helminth parasites are still not a well-known component of animal biodiversity. In this study, we describe and compare the endoparasite assemblages of three native rodent species from a semiarid Mediterranean ecosystem. A total of eight species of gastrointestinal helminth parasites were detected. The endoparasitic communities of the rodents Phyllotis darwini (5 species), Octodon degus (4 species), and Abrothrix olivaceus (3 species) were described in terms of their prevalence and mean abundance. In addition, endoparasite records by rodent species were submitted to rarefaction analyses to verify sampling robustness and used to compare richness and similarity among rodent species. O. degus presented the highest prevalence and parasitic load, but was of low diversity, while P. darwini presented the highest diversity, but had a low prevalence rate. Conversely, A. olivaceus presented the lowest prevalence and diversity. This study contributes to the existing knowledge of the endoparasitic fauna of rodent species from semiarid Mediterranean zones.