Molecular epidemiology of parasitic protozoa and Ehrlichia canis in wildlife in Madrid (central Spain)

Abstract

Wildlife species are involved in the transmission of diverse pathogens. This study aimed to monitor raccoons (Procyon lotor), American minks (Neovison vison), and red foxes (Vulpes vulpes) as potential reservoirs in central Spain. Specifically, 200 spleen and fecal samples (from 194 raccoons, 3 minks, and 3 foxes) were analyzed molecularly by PCR/qPCR and sequencing for the presence of piroplasmids, Hepatozoon spp., Toxoplasma gondii, and Ehrlichia canis infections in the Community of Madrid (Spain). Biological samples were obtained in the years 2014, 2015, and 2016. No pathogen DNA was found in fecal samples. In contrast, analysis of raccoon spleen samples revealed that Toxoplasma was the most prevalent pathogen (prevalence 3.6 ± 2.6%), followed by Hepatozoon canis and E. canis (each with a prevalence of 2.57 ± 2.2%). Hepatozoon canis was also diagnosed in all three of the analyzed foxes. Analysis of yearly prevalence showed that tick-borne pathogens were less frequent in raccoon in 2015, a dry and warm year compared both to 2014 and 2016. These data suggest that fecal PCR assays are unsuitable for detection of DNA of non-erythrocytic pathogens. Furthermore, they demonstrate that the raccoon (an invasive species often living in proximity to domestic areas) and the red fox are putative reservoirs for pathogenic organisms in the Community of Madrid.

Diversity of Anaplasma and Ehrlichia/Neoehrlichia Agents in Terrestrial Wild Carnivores Worldwide: Implications for Human and Domestic Animal Health and Wildlife Conservation

“…Among wild felids, E. canis rrs has been detected in ocelots ( Leopardus pardalis ) (13.3–17.2%), jaguarondis ( Herpailurus yagouaroundi ) (10.5–16.6%), little spotted cats ( Leopardus tigrinus ) (8.0–14.3%), pumas ( Puma concolor ) (1.1–25%), jaguars ( Panthera onca ) (22.2%), lion ( Panthera leo ) (16.6%) maintained in captivity in zoos in Brazil ( 75 , 9 ), free-ranging Iriomote cats (12%) ( Prionailurus iriomotensis ) and Tsushima leopard cats ( Prionailurus bengalensis euptilura ) (8%) in Japan ( 47 ), and lions (1%) maintained in captivity in Zimbabwe ( 42 ) (Table 2 ). Among procyonids and mustelids, E. canis rrs has been detected in raccoons ( Procyon lotor ) (1.7%) from the USA ( 78 ) and Spain (2.6%) ( 36 ), coatis (3.2%) from Brazil ( 12 ), and Eurasian otters (50%) from Italy ( 73 ) (Table 2 ). Although E. canis rrs has been detected in several wild captive felid species in Brazil ( 75 ), phylogenetic analysis based on omp-1 gene positioned Ehrlichia sp.…”

“…Although E. canis has been molecularly detected in free-ranging raccoons from the USA (1.7%) ( 78 ) and Spain (2.6%) ( 36 ), Yabsley et al ( 52 ) showed that Procyon lotor was not susceptible to experimental infection with E. canis . In the USA, although D. variabilis , a recognized vector for E. canis ( 64 ), has been found parasitizing dogs and medium-sized wild mammals, such as raccoons ( 52 , 61 , 78 ), allowing for E. canis inter-species transmission in suburban areas, raccoons seem to play a limited role as a vertebrate reservoir for this agent in the USA ( 78 ).…”

“…In Japan, all 187 raccoons sampled in Kanagawa Prefecture showed to be PCR-negative for E. canis , despite the serological evidence of exposure to this agent in one animal ( 51 ) (Table 2 ). According to Criado-Fornelio et al ( 36 ), E. canis may be tick-transmitted between domestic dogs and wild carnivores, including raccoons, and vice-versa in central Spain. In Brazil, among 31 coatis sampled in the Pantanal wetland, 3.2% showed to be seropositive to E. canis and presented E. canis rrs in blood sample ( 12 ).…”

Invasive alien mammals of European Union concern

“…Overall, studies on Echinococcus multilocularis (14 studies, three species), Toxoplasma gondii (nine studies, six species), and Baylisascaris procyonis (nine studies, one species) were particularly abundant among the study species (Appendix S5). Prevalence rates presented a high geographical and taxonomical variability: the prevalence of E. multilocularis ranged between 0% (in the raccoon and the raccoon dog in various countries; Kornyushin et al 2011, Wahlström et al 2012, EFSA 2015, Oksanen et al 2016, Duscher et al 2017) and 28% (in the racoon dog in Slovakia; Oksanen et al 2016); for T. gondii, it ranged from 0% (in mink in Spain and the raccoon in the Czech Republic; Criado-Fornelio et al 2018, Kornacka et al 2018) to 78.8% (in mink in Spain; Ribas et al 2018); lastly, the prevalence of B. procyonis in raccoons ranged from 1.9% in Poland (Karamon et al 2014) to 80% in Germany (Hohmann et al 2002). Moreover, there are recent reports from Denmark and the Netherlands regarding SARS-CoV-2 infection in minks (Oreshkova et al 2020).…”

“…CC-BY 4.0 International license made available under a (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is The copyright holder for this preprint this version posted April 22, 2021. ; https://doi.org/10.1101/2021.04.21.440832 doi: bioRxiv preprint 13 countries; Kornyushin et al 2011, Wahlström et al 2012, EFSA 2015, Oksanen et al 2016, Duscher et al 2017) and 28% (in the racoon dog in Slovakia; Oksanen et al 2016); for T. gondii, it ranged from 0% (in mink in Spain and the raccoon in the Czech Republic; Criado-Fornelio et al 2018, Kornacka et al 2018) to 78.8% (in mink in Spain; Ribas et al 2018); lastly, the prevalence of B. procyonis in raccoons ranged from 1.9% in Poland (Karamon et al 2014) to 80% in Germany (Hohmann et al 2002). Moreover, there are recent reports from Denmark and the Netherlands regarding SARS-CoV-2 infection in minks (Oreshkova et al 2020).…”