Dinámica poblacional de las garrapatas exófilas en el noroeste de España y su papel como vectores de agentes infecto-parasitarios de importancia veterinaria y médica

Con objeto de conocer las principales especies de garrapatas exófilas y su estacionalidad en el noroeste de España, así como los diferentes agentes patógenos que éstas vehiculan y su dinámica de infección, durante 2 años consecutivos (noviembre del 2015 a octubre del 2017) se realizó, mediante el método del bandereo, una recogida mensual de garrapatas en tres zonas de muestreo (costa, meseta y montaña). Se recogieron 6.956 garrapatas, cuya identificación específica se realizó siguiendo diversas claves morfológicas. Posteriormente, de cada especie de garrapata se seleccionaron 2 ejemplares, considerando el sexo y estadio, para obtener imágenes bajo la lupa y por microscopía electrónica de barrido. Además, varios ejemplares de cada especie y estadio se seleccionaron para confirmar la identificación morfológica mediante el análisis de la secuencia de un fragmento del gen que codifica para la subunidad 16S del ARN ribosómico. Así, se identificaron 5 especies de ixódidos, siendo Ixodes ricinus (99,25%) la especie más abundante, mientras que Dermacentor marginatus (0,27%), Dermacentor reticulatus (0,24%), Ixodes frontalis (0,22%) e Ixodes acuminatus (0,01%) se hallaron de forma ocasional. Además, I. ricinus fue la única especie de garrapata encontrada en todos los estadios de desarrollo (652 larvas, 5.767 ninfas, 227 machos y 258 hembras); sólo se encontraron 1 larva y 14 ninfas de I. frontalis, 36 adultos de Dermacentor (15 hembras y 4 machos de D. marginatus y 10 hembras y 7 machos de D. reticulatus) y un macho adulto de I. acuminatus

Cephenemyia stimulator (Diptera: Oestridae) myiasis in a roe deer (Capreolus capreolus) from Portugal

Herein we present a nasopharyngeal myiasis case by Cephenemyia stimulator in a roe deer hunted in Trás-os-Montes (NE Portugal). Preliminary inspection showed one larva sorting through the nostrils and the examination of the nasopharyngeal cavity showed more than 15 larvae in the glottis and retropharyngeal recesses. Four larvae were collected and stored into 70% ethanol for morphological and molecular identification. Three of the larvae were identified as third instars and the other one as a prepupa of Cephenemyia stimulator, being the first confirmation of this species in roe deer from Portugal. C. stimulator is currently widely distributed in roe deer from central and northern Spain so, the transboundary natural dispersal of these cervids would explain the introduction of this myiasis in Portugal. Further studies are needed to monitor the spreading of this infection in the westernmost populations of European roe deerThe authors are very grateful to Jorge Afonso (Carramo Safaris, Macedo de Cavaleiros, Portugal), the hunter who submitted the larvae for investigation. This work was supported by the projects UIDB7CVT700772/2020 and LA/P/0059/2020 funded by the Portuguese Foundation for Science and Technology (FCT) and by the Program for consolidating and structuring competitive research groups (ED4312019/04, Xunta de Galicia, Spain)S

Molecular characterization of novel Ehrlichia genotypes in Ixodes auritulus from Uruguay

Ehrlichia are small intracellular Gram-negative bacteria transmitted by ticks. These microorganisms cause ehrlichiosis, a complex of life-threatening emerging zoonoses and diseases of global veterinary relevance. The aim of this study was to investigate the presence of Ehrlichia in free-living Ixodes auritulus collected in Uruguay. Ticks were collected from vegetation in five localities from the southeast and northeast of the country between 2014 and 2017. Detection of Ehrlichia DNA was performed in pools of adults or nymphs grouped according to the collection site and date. A total of 1,548 I. auritulus ticks were collected in four of the five locations sampled. Fragments of three loci (16S rRNA, dsb and groEL) were obtained by PCR, and phylogenies inferred using Bayesian inference analysis for each gene independently. DNA of Ehrlichia spp. was found in 15 out of 42 tick pools. Based on the topology of the phylogenetic trees, our sequences represent two novel genotypes for the genus named as Ehrlichia sp. Serrana and Ehrlichia sp. Laguna Negra. Both genotypes were closely related to Ehrlichia sp. Magellanica, a species detected in Ixodes uriae and Magellanic penguins. Considering that all stages of I. auritulus and I. uriae are parasites of birds, their phylogenetic relationships, and common eco-epidemiological profiles, it is reasonable to state that these genotypes of Ehrlichia spp. may represent a natural group likely associated with birds. Our results constitute the first characterization of Ehrlichia spp. in Uruguay. Future studies on birds reported as hosts for I. auritulus are needed to further understand the epidemiological cycles of both Ehrlichia genotypes in the country. Finally, I. auritulus does not feed on humans, so the two Ehrlichia species reported herein might have no implications in human health.EEA RafaelaFil: Félix, María L. Universidad de la República. Facultad de Veterinaria. Laboratorio de Vectores y Enfermedades Transmitidas; UruguayFil: Muñoz-Leal, Sebastian. Universidad de Concepción. Facultad de Ciencias Veterinarias. Departamento de Patología y Medicina Preventiva; ChileFil: Carvalho, Luis A. AgResearch. Grasslands Research Centre; Nueva ZelandaFil: Queirolo, Diego. Universidad de La República. CENUR Noreste; UruguayFil: Remesar Alonso, Susana. Universidade de Santiago de Compostela. Faculty of Veterinary Sciences. Department of Animal Pathology (INVESAGA Group); EspañaFil: Nava, Santiago. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Nava, Santiago. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Armúa-Fernández, María T. Universidad de la República. Facultad de Veterinaria. Laboratorio de Vectores y Enfermedades Transmitidas; UruguayFil: Venzal, José Manuel. Universidad de la República. Facultad de Veterinaria. Laboratorio de Vectores y Enfermedades Transmitidas; Urugua

A case report of fatal feline babesiosis caused by Babesia canis in north western Spain

In Europe, Babesia infections in cats are sporadic and only partial knowledge is currently available since the number of described cases including both the clinical presentation and the molecular identification of the Babesia species involved is limited. In the present case report, the clinical signs, the epidemiological data and the molecular results suggest that this is the first reported fatal case of feline babesiosis caused by Babesia canisThis research was funded by the Program for consolidating and structuring competitive research groups (ED431C 2019/04, Xunta de Galicia, Spain). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscriptS

Prevalence and distribution of Babesia and Theileria species in roe deer from Spain

Babesiosis and Theileriosis are important worldwide-distributed tick-borne diseases for human and animals. Their presence in a particular area depends on the presence of suitable tick-vector and host species as well as competent reservoirs such as roe deer, one of the most abundant wild cervids in Spain. Spleen samples from 174 roe deer hunted in Spain were analysed to determine the prevalence of Babesia and Theileria species. DNA of both piroplasms was firstly detected using a commercial qPCR. Then, positive samples were molecularly characterized at the 18S rRNA and ITS1 genes of Babesia spp. and Theileria spp. The possible influence of some factors such as ecological area, age and sex was also assessed. Overall, 89.7% of roe deer were positive to any of the two piroplasms. Theileria spp. was more prevalent (60.9%) than Babesia spp. (19.0%); species identification could not be achieved in 17.3% of positive samples. Babesia prevalence was significantly higher in young animals and in roe deer from Oceanic regions, in contrast to Theileria spp. Five species were identified: Theileria sp. OT3 (60.3%), Babesia capreoli (15.5%), Babesia venatorum (2.9%), Theileria sp. 3185/02 (0.6%) and Babesia bigemina (0.6%). The coinfection B. capreoli/T. sp. OT3 was the most common (4.6%) followed by B. venatorum/T. sp. OT3 (0.6%) and B. bigemina/T. sp. OT3 (0.6%). Our results reveal that Theileria spp. and Babesia spp. are prevalent piroplasms in roe deer from Spain. These cervids can act as reservoirs for several Babesia and Theileria species, including the zoonotic B. venatorum. This study represents the first description of B. venatorum and B. bigemina in roe deer from Spain.This research was supported by a project grant (2016-CL018) awarded by the Spanish Roe Deer Association [Asociación del Corzo Español (ACE)], the Programme for Consolidating and Structuring Competitive Research Groups (GRC2015/003; Xunta de Galicia, Spain) and the Research Project “RUMIGAL: Rede de estudo multidisciplinar dos ruminantes en Galicia” (R2014/005, Xunta de Galicia, Spain)S

Pedicle myiasis by Lucilia caesar (Diptera, Calliphoridae): an emerging disease in roe deer from north-western Spain

A total of 35 male roe deer (Capreolus capreolus) from Lugo province (north-western Spain) were examined for pedicle myiasis between 2020 and 2022. All these animals had died by different causes and were examined or preserved just after their death to avoid post-mortem infestations. After external and internal head inspection, five animals were diagnosed with severe myiasis at the basis of the antlers (14.29%; 95% confidence interval = 5.38–31.04). All the affected bucks presented a cutaneous wound (1.5–15 cm in diameter) around the pedicles with extensive tissue destruction, exposition of frontal and parietal bones, and massive infestation by dipteran larvae (2–12 mm long). Four of five roe deer showed whitish egg clusters adhered to the basis of the antlers. Moreover, in one animal, six larvae had penetrated the cranial cavity, and in two of them larvae were also found in the nasopharyngeal cavity. All the cases were recorded in summer (May–August). Morphological identification and subsequent molecular confirmation revealed that all animals were infested by different larval stages of Lucilia caesar (Diptera: Calliphoridae). This study represents the first report of pedicle myiasis in Spain. Since this disease is reported sporadically, the detection of five cases in a short period of time suggests an increase in the incidence of this myiasisS

The Goat as a Risk Factor for Parasitic Infections in Ovine Flocks

Small ruminants in northwestern Spain are frequently managed in mixed flocks. Sheep–goat joint management has not been considered a risk factor for parasite infection, so the main objective of this study was to establish if mixed management with goats supposes a risk factor for parasitic infections in ovine flocks. Two thousand and ninety-three sheep were sampled from 74 commercial meat ovine flocks for diagnostic of the main parasites. Goat contact was a risk factor for sheep to be infected by protostrongylids, Dictyocaulus filaria, gastrointestinal nematodes and Eimeria spp. In contrast, Moniezia, Fasciola hepatica, Dicrocoelium dendriticum and Paramphistomidae prevalences were not influenced. Sheep–goat mixed management can be considered as a risk factor, since goats would act as a source of pasture contamination for interspecific parasites (protostrongylids, Dictyocaulus filaria and gastrointestinal nematodes). In relation to host-specific parasites, such as Eimeria spp., goats cannot be considered as a source for sheep, but competition for food and spaces between both small ungulates can suppose a reduced grazing area to sheep, provoking high environmental contamination and stress that facilitate their infection. Future epidemiological studies for parasitic infections in small ruminants should consider sheep–goat mixed management as a possible risk factor to be included in multivariate analysesThis work was supported by a program for consolidating and structuring competitive research groups (ED431D 2019/04, Xunta de Galicia, Spain)S

Specificity of Stenurus (Metastrongyloidea: Pseudaliidae) infections in odontocetes stranded along the north-west Spanish coast

Parasites extracted from the lungs and the pterygoid sinus complex of 6 species of odontocetes stranded along the north-west Spanish coast (Northeast Atlantic) between 2009 and 2019 were morphologically identified. The samples belonged to 14 specimens, including 3 harbour porpoises, Phocoena phocoena, 6 short-finned pilot whales, Globicephala macrorhynchus, 1 long-finned pilot whale, Globicephala melas, 1 Risso's dolphin, Grampus griseus, 1 striped dolphin, Stenella coeruleoalba and 2 bottlenose dolphins, Tursiops truncatus. All animals (14/14) were infected by nematodes of the genus Stenurus spp.; moreover, two of them presented a mixed lung nematode infection by Stenurus spp. and Halocercus spp., and another two a mixed infection by Stenurus spp. and the trematode Nasitrema spp. in the pterygoid sinuses. The morphological characterization of the Stenurus specimens revealed the existence of three different species: Stenurus minor, present in the pterygoid sinuses of harbour porpoises with a mean intensity of 43.0 ± 9.0; Stenurus globicephalae, in the pterygoid sinuses of pilot whales and the Risso's dolphin (370.3 ± 579.4); and Stenurus ovatus infecting bottlenose and striped dolphins’ lungs (47.7 ± 76.5). This is the first citation of S. minor and S. ovatus in odontoceti from the Galician coast. Nematodes of the genus Stenurus are frequent in odontocetes stranded along the north-west Spanish coast. A clear host-parasite association was observed between S. minor and the Phocoenidae family, between S. globicephalae and the subfamily Globicephalinae and between S. ovatus and subfamily Delphininae. Different trophic position and niche segregation may lead to different patterns of specificityS

Cardiorespiratory nematodes and co-infections with gastrointestinal parasites in new arrivals at dog and cat shelters in north-western Spain

Metastrongyloid and trichuroid cardiorespiratory nematodes of dogs and cats are considered emergent in several European countries, and an increase in the number and extent of endemic foci has been described. Since data on their prevalence are limited in this continent, faecal samples from new arrivals (365 dogs and 65 cats) at two animal shelters in North-western Spain were analysed using both floatation and Baermann techniques. In order to confirm the microscopic identification of Metastrongylidae first stage larvae, molecular characterization based on the sequence of the ITS-2 was performed. The possible influence of some variables such as the species, sex and age of the animals and the co-infection with other gastrointestinal parasites on the prevalence of cardiorespiratory nematodes was analysed. The most prevalent metastrongylid was Aelurostrongylus abstrusus (15.4%) followed by Angiostrongylus vasorum (4.1%) and Crenosoma vulpis (1.1%). Regarding trichuroids, Eucoleus aerophilus and/or Eucoleus boehmi eggs were detected in 28 dogs (7.7%) and four cats (6.2%). Almost all animals positive to cardiorespiratory nematodes (86.8%) were co-infected with gastrointestinal parasites. The prevalence of Metastrongylidae and respiratory trichuroids was significantly higher in dogs co-infected with Taenia spp. and Toxocara canis or Giardia duodenalis and Sarcocystis spp., respectively. In cats, a significant higher prevalence of Metastrongylidae nematodes was found in animals co-infected with Toxocara cati. Our results reveal that cardiorespiratory nematodes are common in companion animals from north-western Spain, showing higher prevalences than those previously reported from this country. This investigation represents the first report of C. vulpis, E. aerophilus and E. boehmi in dogs from Spain. The identification of a number of zoonotic parasites is of public health concern. Our results indicate that these nematodes should be included in the differential diagnosis of dogs and cats from north-western Spain showing respiratory or cardiac clinical signsThis research was funded by the Program for consolidating and structuring competitive research groups (ED431C 2019/04, Xunta de Galicia, Spain)S

The Age-Related Cryptosporidium Species Distribution in Asymptomatic Cattle from North-Western Spain

An age-related distribution of Cryptosporidium species has been reported in cattle, with C. parvum being predominant in suckling calves, C. bovis and C. ryanae being predominant in post-weaned calves and C. andersoni being predominant in adults. However, variants to this pattern have recently been reported. Thus, fecal samples (n = 594) from asymptomatic cattle were collected in north-western Spain. Animals were classified as 2 years (G5). Cryptosporidium detection and species identification were performed by SSU rRNA PCR. Individual Cryptosporidium prevalence was 16.7%; it significantly decreased with age. Cryptosporidium parvum was predominant in G1 and C. bovis was predominant in the rest of the age classes; C. bovis and C. ryanae were especially prevalent in G2 and G3. Cryptosporidium occultus was not found in suckling calves. Finally, C. andersoni and C. xiaoi were occasionally detected in G5. The presence of C. parvum in all age classes implies significant animal and public health concerns. The predominance of C. bovis in cattle older than 1 month supports the idea that the age-related pattern of Cryptosporidium species described in cattle is not fully consistent, and thus further studies are still needed to identify those factors determining the species distributionThis research was funded by Ministerio de Ciencia e Innovación (Spain), grant number AGL2016-76034-P and by Xunta de Galicia (Spain), grant number 2017-PG117S