RESUMEN
The increasing frequency and cost of zoonotic disease emergence due to global change have led to calls for the primary surveillance of wildlife. This should be facilitated by the ready availability of remotely sensed environmental data, given the importance of the environment in determining infectious disease dynamics. However, there has been little evaluation of the temporal predictiveness of remotely sensed environmental data for infection reservoirs in vertebrate hosts due to a deficit of corresponding high-quality long-term infection datasets. Here we employ two unique decade-spanning datasets for assemblages of infectious agents, including zoonotic agents, in rodents in stable habitats. Such stable habitats are important, as they provide the baseline sets of pathogens for the interactions within degrading habitats that have been identified as hotspots for zoonotic emergence. We focus on the enhanced vegetation index (EVI), a measure of vegetation greening that equates to primary productivity, reasoning that this would modulate infectious agent populations via trophic cascades determining host population density or immunocompetence. We found that EVI, in analyses with data standardised by site, inversely predicted more than one-third of the variation in an index of infectious agent total abundance. Moreover, in bipartite host occupancy networks, weighted network statistics (connectance and modularity) were linked to total abundance and were also predicted by EVI. Infectious agent abundance and, perhaps, community structure are likely to influence infection risk and, in turn, the probability of transboundary emergence. Thus, the present results, which were consistent in disparate forest and desert systems, provide proof-of-principle that within-site fluctuations in satellite-derived greenness indices can furnish useful forecasting that could focus primary surveillance. In relation to the well-documented global greening trend of recent decades, the present results predict declining infection burden in wild vertebrates in stable habitats; but if greening trends were to be reversed, this might magnify the already upwards trend in zoonotic emergence.
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Ecosistema , Roedores , Animales , Animales Salvajes , BosquesRESUMEN
In June 2023, a fatal disease outbreak in cats occurred in Poland. Most cases tested in Poland (29 of 47) were positive for highly pathogenic avian influenza (HPAI) A (H5N1) virus. Genetic analyses revealed clade 2.3.4.4b with point mutations indicative of initial mammalian hosts adaptations. Cat viral sequences were highly similar (n = 21), suggesting a potential common infection source. To investigate possible infection routes, our group tested food samples from affected households. HPAI H5N1 virus was detected in one poultry meat sample.
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Subtipo H5N1 del Virus de la Influenza A , Virus de la Influenza A , Gripe Aviar , Animales , Gatos , Subtipo H5N1 del Virus de la Influenza A/genética , Gripe Aviar/epidemiología , Polonia/epidemiología , Aves , Filogenia , MamíferosRESUMEN
We report results from serologic surveillance for exposure to SARS-CoV-2 among 1,237 wild rodents and small mammals across Europe. All samples were negative, with the possible exception of 1. Despite suspected potential for human-to-rodent spillover, no evidence of widespread SARS-CoV-2 circulation in rodent populations has been reported to date.Esitämme tulokset serologisesta tutkimuksesta, jossa seulottiin SARS-CoV-2 tartuntojen varalta 1,237 luonnonvaraista jyrsijää ja piennisäkästä eri puolilta Eurooppaa. Kaikki näytteet olivat negatiivisia, yhtä näytettä lukuun ottamatta. SARS-CoV-2:n läikkymisen ihmisistä jyrsijöihin on arveltu olevan mahdollista, mutta todisteet viruksen laajamittaisesta leviämisestä jyrsijäpopulaatioissa puuttuvat.
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COVID-19 , Animales , Humanos , COVID-19/epidemiología , SARS-CoV-2 , Roedores , Anticuerpos Antivirales , Europa (Continente)/epidemiologíaRESUMEN
Major histocompatibility complex (MHC) genes encode proteins crucial for adaptive immunity of vertebrates. Negative frequency-dependent selection (NFDS), resulting from adaptation of parasites to common MHC types, has been hypothesized to maintain high, functionally relevant polymorphism of MHC, but demonstration of this relationship has remained elusive. In particular, differentiation of NFDS from fluctuating selection, resulting from changes in parasite communities in time and space (FS), has proved difficult in short-term studies. Here, we used temporal data, accumulated through long-term monitoring of helminths infecting bank voles (Myodes glareolus), to test specific predictions of NFDS on MHC class II. Data were collected in three, moderately genetically differentiated subpopulations in Poland, which were characterized by some stable spatiotemporal helminth communities but also events indicating introduction of new species and loss of others. We found a complex association between individual MHC diversity and species richness, where intermediate numbers of DRB supertypes correlated with lowest species richness, but the opposite was true for DQB supertypes-arguing against universal selection for immunogenetic optimality. We also showed that particular MHC supertypes explain a portion of the variance in prevalence and abundance of helminths, but this effect was subpopulation-specific, which is consistent with both NFDS and FS. Finally, in line with NFDS, we found that certain helminths that have recently colonized or spread in a given subpopulation, more frequently or intensely infected voles with MHC supertypes that have been common in the recent past. Overall, our results highlight complex spatial and temporal patterns of MHC-parasite associations, the latter being consistent with Red Queen coevolutionary dynamics.
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Arvicolinae , Helmintos , Animales , Arvicolinae/genética , Helmintos/genética , Antígenos de Histocompatibilidad Clase II/genética , Polonia , Polimorfismo Genético , Selección GenéticaRESUMEN
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiologic agent of coronavirus disease and has been spreading worldwide since December 2019. The virus can infect different animal species under experimental conditions, and mink on fur farms in Europe and other areas are susceptible to SARS-CoV-2 infection. We investigated SARS-CoV-2 infection in 91 mink from a farm in northern Poland. Using reverse transcription PCR, antigen detection, and next-generation sequencing, we confirmed that 15 animals were positive for SARS-CoV-2. We verified this finding by sequencing full viral genomes and confirmed a virus variant that has sporadic mutations through the full genome sequence in the spike protein (G75V and C1247F). We were unable to find other SARS-CoV-2 sequences simultaneously containing these 2 mutations. Country-scale monitoring by veterinary inspection should be implemented to detect SARS-CoV-2 in other mink farms.
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COVID-19 , Visón , Animales , Granjas , Humanos , Polonia/epidemiología , SARS-CoV-2RESUMEN
Routine genomic surveillance on samples from COVID-19 patients collected in Poland during summer 2021 revealed the emergence of a SARS-CoV-2 Delta variant with a large 872 nt deletion. This change, confirmed by Sanger and deep sequencing, causes complete loss of ORF7a, ORF7b, and ORF8 genes. The index case carrying the deletion is unknown. The standard pipeline for sequencing may mask this deletion with a long stretch of N's. Effects of this deletion on phenotype or immune evasion needs further study.
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COVID-19 , SARS-CoV-2 , Humanos , PoloniaRESUMEN
Bank voles in Poland are reservoirs of zoonotic viruses. To determine seroprevalence of hantavirus, arenavirus, and cowpox virus and factors affecting seroprevalence, we screened for antibodies against these viruses over 9 years. Cowpox virus was most prevalent and affected by extrinsic and intrinsic factors. Long-term and multisite surveillance is crucial.
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Arvicolinae/virología , Enfermedades de los Roedores/epidemiología , Enfermedades de los Roedores/virología , Animales , Estudios Transversales , Femenino , Historia del Siglo XXI , Masculino , Polonia/epidemiología , Estudios Seroepidemiológicos , ZoonosisRESUMEN
The CRISPR/Cas9 system, a natural defence system of bacterial organisms, has recently been used to modify genomes of the most important protozoa parasites. Successful genome manipulations with the CRISPR/Cas9 system are changing the present view of genetics in parasitology. The application of this system offers a major chance to overcome the current restriction in culturing, maintaining and analysing protozoan parasites, and allows dynamic analysis of parasite genes functions, leading to a better understanding of pathogenesis. CRISPR/Cas9 system will have a significant influence on the process of developing novel drugs and treatment strategies against protozoa parasites.
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Sistemas CRISPR-Cas , Eucariontes/fisiología , Parásitos/fisiología , Animales , Eucariontes/genética , Genoma de Protozoos/genética , Parásitos/genéticaRESUMEN
The reprogramming of human induced pluripotent stem cells (hiPSCs) proceeds in a stepwise manner with reprogramming factors binding and epigenetic composition changes during transition to maintain the epigenetic landscape, important for pluripotency. There arises a question as to whether the aberrant epigenetic state after reprogramming leads to epigenetic defects in induced stem cells causing unpredictable long term effects in differentiated cells. In this review, we present a comprehensive view of epigenetic alterations accompanying reprogramming, cell maintenance and differentiation as factors that influence applications of hiPSCs in stem cell based technologies. We conclude that sample heterogeneity masks DNA methylation signatures in subpopulations of cells and thus believe that beside a genetic evaluation, extensive epigenomic screening should become a standard procedure to ensure hiPSCs state before they are used for genome editing and differentiation into neurons of interest. In particular, we suggest that exploitation of the single-cell composition of the epigenome will provide important insights into heterogeneity within hiPSCs subpopulations to fast forward development of reliable hiPSC-based analytical platforms in neurological disorders modelling and before completed hiPSC technology will be implemented in clinical approaches.
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Reprogramación Celular/genética , Reprogramación Celular/fisiología , Epigénesis Genética , Enfermedades del Sistema Nervioso/genética , Enfermedades del Sistema Nervioso/fisiopatología , Animales , Epigenómica , Humanos , Células Madre Pluripotentes/fisiologíaRESUMEN
UNLABELLED: A significant number of studies report growing resistance in nematodes thriving in both humans and livestock. This study was conducted to evaluate the in vitro and in vivo anthelmintic efficiency of Curcubita pepo (C. pepo) L. hot water extract (HWE), cold water extract (CWE) or ethanol extract (ETE) on two model nematodes: Caenorhabditis elegans (C. elegans) and Heligmosoides bakeri (H. bakeri). METHODS: Raman, IR and LC-MS spectroscopy analyses were performed on the studied plant material to deliver qualitative and quantitative data on the composition of the obtained extracts: ETE, HWE and CWE. The in vitro activity evaluation showed an impact of C. pepo extracts on C. elegans and different developmental stages of H. bakeri. The following in vivo experiments on mice infected with H. bakeri confirmed inhibitory properties of the most active pumpkin extract selected by the in vitro study. All of the extracts were found to contain cucurbitine, aminoacids, fatty acids, and-for the first time-berberine and palmatine were identified. All C. pepo seed extracts exhibited a nematidicidal potential in vitro, affecting the survival of L1 and L2 H. bakeri larvae. The ETE was the strongest and demonstrated a positive effect on H. bakeri eggs hatching and marked inhibitory properties against worm motility, compared to a PBS control. No significant effects of pumpkin seed extracts on C. elegans integrity or motility were found. The EtOH extract in the in vivo studies showed anthelmintic properties against both H. bakeri fecal egg counts and adult worm burdens. The highest egg counts reduction was observed for the 8 g/kg dose (IC50 against H. bakeri = 2.43; 95% Cl = 2.01-2.94). A decrease in faecal egg counts (FEC) was accompanied by a significant reduction in worm burden of the treated mice compared to the control group. CONCLUSIONS: Pumpkin seed extracts may be used to control of Gastrointestinal (G.I.) nematode infections. This relatively inexpensive alternative to the currently available chemotherapeutic should be considered as a novel drug candidate in the nearest future.
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Antinematodos/farmacología , Cucurbita/química , Extractos Vegetales/farmacología , Semillas/química , Aminoácidos/análisis , Animales , Antinematodos/química , Berberina/análisis , Alcaloides de Berberina/análisis , Ácidos Grasos/análisis , Ratones , Extractos Vegetales/química , Proteínas de Plantas/análisis , Rabdítidos/efectos de los fármacosRESUMEN
Parasites are considered to be an important selective force in host evolution but ecological studies of host-parasite systems are usually short-term providing only snap-shots of what may be dynamic systems. We have conducted four surveys of helminths of bank voles at three ecologically similar woodland sites in NE Poland, spaced over a period of 11 years, to assess the relative importance of temporal and spatial effects on helminth infracommunities. Some measures of infracommunity structure maintained relative stability: the rank order of prevalence and abundance of Heligmosomum mixtum, Heligmosomoides glareoli and Mastophorus muris changed little between the four surveys. Other measures changed markedly: dynamic changes were evident in Syphacia petrusewiczi which declined to local extinction, while the capillariid Aonchotheca annulosa first appeared in 2002 and then increased in prevalence and abundance over the remaining three surveys. Some species are therefore dynamic and both introductions and extinctions can be expected in ecological time. At higher taxonomic levels and for derived measures, year and host-age effects and their interactions with site are important. Our surveys emphasize that the site of capture is the major determinant of the species contributing to helminth community structure, providing some predictability in these systems.
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Arvicolinae/parasitología , Helmintiasis Animal/epidemiología , Enfermedades de los Roedores/epidemiología , Factores de Edad , Animales , Biodiversidad , Estudios Transversales , Femenino , Helmintiasis Animal/parasitología , Helmintos/crecimiento & desarrollo , Intestinos/parasitología , Masculino , Polonia/epidemiología , Prevalencia , Enfermedades de los Roedores/parasitología , Análisis Espacio-TemporalRESUMEN
Abundance and prevalence of helminth infections often differ between host sexes, and are usually biased in favor of males. Relatively few cases of female-biased parasitism have been reported. We sampled bank voles in three woodland sites in N.E. Poland over 11 years at 3-4-year intervals, and assessed their parasite burdens. Prevalence and abundance of the stomach nematode Mastophorus muris were consistently higher among females. Among adult female bank voles from the two sites that showed the highest prevalence with M. muris, both prevalence and abundance were significantly higher in lactating bank voles, but not pregnant animals, and the effect of lactation was evident in both sites, in all four surveys, and in both age classes. Although the magnitude of the effect of lactation varied between years, it was not confounded by any significant interactions with other factors. We hypothesize that mature and reproductively active female bank voles are subject to higher exposure compared with males of similar age, as a consequence of the increased content of invertebrates in their diet, including the intermediate hosts of M. muris, required to meet the higher increased energy and protein demands of nursing litters throughout the summer months.
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Arvicolinae/parasitología , Parasitosis Intestinales/veterinaria , Enfermedades de los Roedores/epidemiología , Infecciones por Spirurida/veterinaria , Spiruroidea/fisiología , Animales , Femenino , Humanos , Parasitosis Intestinales/epidemiología , Parasitosis Intestinales/parasitología , Lactancia , Masculino , Polonia/epidemiología , Embarazo , Complicaciones Parasitarias del Embarazo/epidemiología , Complicaciones Parasitarias del Embarazo/parasitología , Complicaciones Parasitarias del Embarazo/veterinaria , Prevalencia , Enfermedades de los Roedores/parasitología , Estaciones del Año , Factores Sexuales , Infecciones por Spirurida/epidemiología , Infecciones por Spirurida/parasitología , Spiruroidea/aislamiento & purificación , Estómago/parasitologíaRESUMEN
For this study, 724 gastrointestinal tracts of slaughter horses were investigated to determine the prevalence, intensity of Anoplocephala perfoliata and tapeworm development stages over the second, third and fourth quarter of 2012 and the first quarter of 2013. For each positive horse, faecal samples were collected from the rectum or small colon for coproscopic examinations. The samples were analysed using dedicated modified sedimentation-flotation methods. In total, 52 horses were infected with A. perfoliata in the course of the study, with an overall prevalence of 7.2 %. The prevalence changed over the study period; however, not markedly. The overall mean of A. perfoliata abundance was 12.3 (3.23) and did not differ significantly between the quarters. Mean invasion intensity did not differ significantly between the quarters. The quantity of mature tapeworms did not differ significantly over the study period; however, there was a significant difference in the number of immature tapeworms. The highest number of mature tapeworms was found in the first quarter of 2013. The number of detected tapeworm eggs rose significantly over the study period. The total number of tapeworms did not have a significant influence on the presence/absence of detected eggs. However, there was a noticeable difference between the number of mature tapeworms and presence/absence of eggs in faeces. This clearly indicates that the efficacy of the modified sedimentation-flotation method is influenced by seasonality, and therefore the most effective and reliable time for detection of A. perfoliata in equines is the first quarter of the year.
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Cestodos/aislamiento & purificación , Infecciones por Cestodos/veterinaria , Enfermedades de los Caballos/diagnóstico , Parasitosis Intestinales/veterinaria , Animales , Cestodos/crecimiento & desarrollo , Infecciones por Cestodos/diagnóstico , Infecciones por Cestodos/epidemiología , Infecciones por Cestodos/parasitología , Clima , Heces/parasitología , Enfermedades de los Caballos/epidemiología , Enfermedades de los Caballos/parasitología , Caballos , Parasitosis Intestinales/diagnóstico , Parasitosis Intestinales/epidemiología , Parasitosis Intestinales/parasitología , Recuento de Huevos de Parásitos , Polonia/epidemiología , Prevalencia , Estaciones del Año , TemperaturaRESUMEN
The autopsy of 487 slaughter horses revealed the presence of Anoplocephala perfoliata in 36 animals. The invasions varied in the intensity (3 to 2,069 tapeworms) and in the level of tapeworms' proglottid maturity. Twenty nine horses were found to contain tapeworms with gravid proglottid. Fecal samples collected from the rectum were tested using following techniques: flotation with solution-saturated NaCl, decantation, McMaster's, and modified sedimentation-flotation methods (50 g feces samples, flotation solution-saturated NaCl and sucrose, specific gravity 1.25 g/ml). The number of A. perfoliata positive fecal samples was significantly higher using the sedimentation-flotation methods 21 (58.33%) than flotation 6 (16.66%), decantation 3 (8.33%), and McMaster's 1 (2.77%) techniques. The sensitivities of the coprological methods during the patent period were 20.69, 10.34, 3.45, and 72.41% for the flotation, decantation, McMaster's, and sedimentation-flotation method, respectively. Sedimentation-flotation techniques proved to be more sensitive than other one. The lowest intensity of invasion possible to detect using this method was nine tapeworms with gravid proglottid.
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Cestodos/aislamiento & purificación , Infecciones por Cestodos/veterinaria , Heces/parasitología , Enfermedades de los Caballos/parasitología , Animales , Infecciones por Cestodos/diagnóstico , Infecciones por Cestodos/parasitología , Enfermedades de los Caballos/diagnóstico , Caballos , Sensibilidad y EspecificidadRESUMEN
By 2030, an estimated 2 billion international tourist trips are expected annually worldwide, with citizens of Poland as important contributors. Illness rates among returnees from developing regions range between 43% and 79%. Properly diagnosing fever in these travelers is vital due to potentially serious implications. After visiting tropical and subtropical zones, the main health complaints are diarrhea, fever, and skin lesions. A reliable diagnosis begins with taking a comprehensive travel history and identifying potential risks. In travelers returning from subSaharan Africa, malaria caused by Plasmodium falciparum is the main cause of fever, affecting 50 in every 1000 cases. Among returnees from Southeast Asia, dengue is dominant, occurring in 50-60 per 1000 cases, and its prevalence rises significantly nowadays. Other significant diseases include chikungunya, Zika, typhoid fever, amebic liver abscess, and occasionally viral hemorrhagic fevers. SARSCoV2 and influenza viruses are crucial pathogens as well. An indepth assessment of the travel history, combined with knowledge on tropical diseases, are key to the diagnostic process, and algorithms may be helpful in selecting appropriate tests and treatment methods.
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Malaria , Fiebre Tifoidea , Infección por el Virus Zika , Virus Zika , Humanos , Malaria/complicaciones , Malaria/diagnóstico , Malaria/epidemiología , Fiebre/etiología , Fiebre Tifoidea/complicaciones , Fiebre Tifoidea/diagnóstico , Fiebre Tifoidea/epidemiología , Viaje , Polonia , Infección por el Virus Zika/complicacionesRESUMEN
BACKGROUND: Biomonitoring is an essential activity for identifying possible vectors and reservoirs of pathogens and predicting potential outbreaks. Wild red foxes are present in both sylvatic and synanthropic environments, making them potential carriers of zoonotic pathogens. Experimental studies have shown that both coyotes and red foxes can transmit SARS-CoV-2. This study aimed to assess the prevalence and seroprevalence of SARS-CoV-2 in wild red foxes hunted in northern Poland. METHODS: Oral swabs, blood clots or heat tissue samples were collected from 292 red foxes hunted in northern Poland. We used both molecular (RT-PCR) and serological (IFA) approaches to detect SARS-CoV-2 infections in the sampled animals. RESULTS: We did not find any evidence of SARS-CoV-2 infection in the collected samples, using both molecular and serological methods. CONCLUSIONS: Despite foxes having frequent contact with humans, human waste, and other animals, they do not appear to participate in the circulation of the SARS-CoV-2 virus in our geographical region. Nevertheless, we believe that continuous biomonitoring should be performed to assess the SARS-CoV-2 epidemiological situation in the wild.
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Background: Our study explores the role of bats as reservoirs of coronaviruses. Methods: We conducted virological screening of bats hibernating in military bunkers at the Natura 2000 site "Nietoperek" in Western Poland collecting oral and anal swab samples from 138 bats across six species to apply a combination of pan-coronavirus and SARS-CoV-2 specific PCR assays. Results: Only one anal swab tested positive for coronavirus. No SARS-CoV-2 was detected in any of the samples. The low prevalence of coronavirus in the studied colony contrasts with higher rates found in other regions and may be influenced by hibernation. Conclusions: Hibernating bats may show a low prevalence of coronavirus, potentially due to the hibernation process itself. This finding indicates that hibernating bats may not be the most optimal subjects for screening zoonotic pathogens. However, biomonitoring of bats for emerging and reemerging diseases is recommended for comprehensive epidemiological insights.
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In an age defined by rapid globalization and unprecedented technological advancements, the field of infectious diseases stands at the intersection of complex challenges and promising opportunities [...].
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Enfermedades Transmisibles , Humanos , Enfermedades Transmisibles/epidemiologíaRESUMEN
Candidatus Neoehrlichia mikurensis (CNM) and Hepatozoon spp. are important vector-borne parasites of humans and animals. CNM is a relatively recently discovered pathogen of humans. Hepatozoon are parasites of reptiles, amphibians and mammals, commonly found in rodents and carnivores worldwide. The present study aimed to determine the prevalence of CNM and Hepatozoon spp. in three species of Microtus and to assess the occurrence of vertical transmission in naturally-infected voles. Molecular techniques were used to detect pathogen DNA in blood and tissue samples of captured voles and their offspring. The prevalence of CNM in the vole community ranged 24-47% depending on Microtus species. The DNA of CNM was detected in 21% of pups from three litters of six infected Microtus dams (two Microtus arvalis and one M. oeconomus) and in 3/45 embryos (6.6%) from two litters of eight CNM-infected pregnant females. We detected Hepatozoon infection in 14% of M. arvalis and 9% of M. oeconomus voles. Hepatozoon sp. DNA was detected in 48.7% of pups from seven litters (6 M. arvalis and 1 M. oeconomus) and in two embryos (14.3%) obtained from one M. arvalis litter. The high prevalence of CNM infections in the Microtus spp. community may be a result of a relatively high rate of vertical transmission among naturally infected voles. Vertical transmission was also demonstrated for Hepatozoon sp. in M. arvalis and M. oeconomus. Our study underlines the significance of alternative routes of transmission of important vector-borne pathogens.
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Anaplasmataceae , Transmisión Vertical de Enfermedad Infecciosa , Animales , Femenino , Humanos , Embarazo , Transmisión Vertical de Enfermedad Infecciosa/veterinaria , ArvicolinaeRESUMEN
Small mammals are suspected of contributing to the dissemination of Toxocara canis and helping with the parasite survival during periods when there is a temporary absence of suitable definitive hosts. While the primary aim of the current study was the assessment of seroprevalence of Toxocara spp. infections in wild rodents in Poland, we also explored the role of intrinsic (sex, age) and extrinsic factors (study site) influencing dynamics of this infection to ascertain whether grassland versus forest rodents play a greater role as indicators of environmental contamination with T. canis. We trapped 577 rodents belonging to four species (Myodes glareolus, Microtus arvalis, Microtus agrestis, Alexandromys oeconomus) in north-eastern Poland. Blood was collected during the parasitological examination, and serum was frozen at - 80 °C until further analyses. A bespoke enzyme-linked immunosorbent assay was used to detect antibodies against Toxocara spp. We found Toxocara spp. antibodies in the sera of all four rodent species with an overall seroprevalence of 2.8% [1.9-4.1%]. There was a significant difference in seroprevalence between vole species, with the grassland species (M. arvalis, M. agrestis and A. oeconomus) showing a 16-fold higher seroprevalence (15.7% [8.7-25.9%]) than the forest-dwelling M. glareolus (0.98% [0.5-1.8%]). We hypothesise that the seroprevalence of Toxocara spp. differs between forest and grassland rodents because of the higher contamination of grasslands by domestic dogs and wild canids. Our results underline the need for wide biomonitoring of both types of ecosystems to assess the role of rodents as indicators of environmental contamination with zoonotic pathogens.