Browsing by Subject " Genotyping "

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    Free-Ranging Wolves (Canis lupus) are Natural Reservoirs of Intestinal Microeukaryotes of Public Health Significance in Southwestern Europe
    (Wiley, 2024-12-08) Ortega, Sheila; Figueiredo, Ana M.; Moroni, Barbara; Abarca, Nadia; Dashti, Alejandro; Köster, Pamela C.; Bailo, Begoña; Cano-Terriza, David; Fayos, Manena; Oleaga, Álvaro; Martínez-Carrasco Pleite, Carlos; Velarde, Roser; Torres, Rita T.; Ferreira, Eduardo; Hipólito, Dário; Barros, Tânia; Lino, Ana; Robetto, Serena; Rossi, Luca; Muñoz-de-Mier, Gemma J.; Ávalos, Gabriel; Calero- Bernal, Rafael; González- Barrio, David; Sánchez, Sergio; García- Bocanegra, Ignacio; Carmena, David; Gonzálvez Juan, Moisés; Sanidad Animal; Facultad de Veterinaria
    Introduction: Intestinal microeukaryote parasites are major contributors to the burden of diarrhoea in humans and domestic animals, but their epidemiology in wildlife is not fully understood. We investigated the frequency, genetic diversity and zoonotic potential of protists of animal and public health significance in free-ranging grey wolf (Canis lupus) populations in south-western Europe. Methods: Individually formed faecal samples collected from necropsied wolves or scat trails in Italy (n = 47), Portugal (n = 43) and Spain (n = 225) during the period 2011–2023 were retrospectively analysed using molecular (PCR and Sanger sequencing) methods. Complementary epidemiological data were gathered when available. Results: Giardia duodenalis was the most frequent microeukaryote found (40.3%, 127/315; 95% CI: 34.9–46.0), followed by Cryptosporidium spp. (3.5%, 11/315; 95% CI: 1.8–6.2), Enterocytozoon bieneusi and Encephalitozoon spp. (1.6%, 5/315; 95% CI: 0.5–3.7 each). Blastocystis was not identified in any of the faecal samples analysed. Sequence analyses confirmed the presence of canine-adapted assemblage D within G. duodenalis (n = 7). Three Cryptosporidium species were identified, namely canine-adapted C. canis (n = 9), zoonotic C. parvum (n = 1) and primarily anthroponotic C.hominis (n = 1). Genotyping tools enabled the identification of subtype family XXe2 within C. canis. Among microsporidia, the canine-adapted genotype PtEb IX was identified within E. bieneusi. Two samples were confirmed as Enc. intestinalis and three more as Enc. cuniculi genotype IV. This is the first record of Enc. intestinalis and Enc. cuniculi in the grey wolf globally. Conclusions: Silent carriage of intestinal microeukaryotes seems common in free-ranging grey wolves in southwestern Europe. Wolves can contribute to environmental contamination through the transmission stages (cysts, oocysts, spores) of species/genotypes potentially infective to humans. Individuals in close contact with wolf carcasses or their faecal material may be at potential risk of infection by microeukaryotic pathogens.
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    Occurrence and limited zoonotic potential of Cryptosporidium spp., Giardia duodenalis, and Balantioides coli infections in free-ranging and farmed wild ungulates in Spain
    (Elsevier, 2023-04-27) Dashti, Alejandro; Köster, Pamela C.; Bailo, Begoña; Sánchez de las Matas, Ana; Habela, Miguel Ángel; Rivero-Juarez, Antonio; Vicente Ibáñez, Joaquín; Serrano, Emmanuel; Arnal, María C.; Fernández de Luco, Daniel; Morrondo, Patrocinio; Armenteros, José A.; Balseiro, Ana; Cardona, Guillermo A.; Martínez-Carrasco Pleite, Carlos; Ortiz, José Antonio; Carpio, Antonio José; Calero Bernal, Rafael; González-Barrio, David; Carmena, David; Sanidad Animal; Facultad de Veterinaria
    Little information is currently available on the occurrence and molecular diversity of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Balantioides coli in wild ungulates and the role of these host species as potential sources of environmental contamination and consequent human infections. The presence of these three pathogens was investigated in eight wild ungulate species present in Spain (genera Ammotragus, Capra, Capreolus, Cervus, Dama, Ovis, Rupicapra, and Sus) by molecular methods. Faecal samples were retrospectively collected from free-ranging (n = 1058) and farmed (n = 324) wild ungulates from the five Spanish bioregions. Overall infection rates were 3.0% (42/1382; 95% CI: 2.1–3.9%) for Cryptosporidium spp., 5.4% (74/1382; 95% CI: 4.2–6.5%) for G. duodenalis, and 0.7% (9/1382; 95% CI: 0.3–1.2%) for B. coli. Cryptosporidium infection was detected in roe deer (7.5%), wild boar (7.0%) and red deer (1.5%), and G. duodenalis in southern chamois (12.9%), mouflon (10.0%), Iberian wild goat (9.0%), roe deer (7.5%), wild boar (5.6%), fallow deer (5.2%) and red deer (3.8%). Balantioides coli was only detected in wild boar (2.5%, 9/359). Sequence analyses revealed the presence of six distinct Cryptosporidium species: C. ryanae in red deer, roe deer, and wild boar; C. parvum in red deer and wild boar; C. ubiquitum in roe deer; C. scrofarum in wild boar; C. canis in roe deer; and C. suis in red deer. Zoonotic assemblages A and B were detected in wild boar and red deer, respectively. Ungulateadapted assemblage E was identified in mouflon, red deer, and southern chamois. Attempts to genotype samples positive for B. coli failed. Sporadic infections by canine- or swine-adapted species may be indicative of potential cross-species transmission, although spurious infections cannot be ruled out. Molecular evidence gathered is consistent with parasite mild infections and limited environmental contamination with (oo)cysts. Free-ranging wild ungulate species would not presumably play a significant role as source of human infections by these pathogens. Wild ruminants do not seem to be susceptible hosts for B. coli.