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Background: Paramphistomids are ruminal trematodes that cause important losses in tropical and subtropical regions. However, their presence in Europe has increased significantly in recent decades. In northwestern Spain, this trend has been confirmed in cattle, but data in sheep are scarce and not updated. Moreover, the paramphistomid species affecting sheep in the area have never been molecularly identified. To evaluate the prevalence of paramphistomid infections in sheep from northwestern Spain, 826 faecal samples from 25 sheep farms were collected and analysed via coproscopic techniques. In addition, the rumens of 85 roe deer from the same area were examined to detect adult paramphistomids. The species present were molecularly identified. Multivariate analyses for identifying the risk factors affecting the prevalence and egg shedding of rumen flukes were also performed., Results: Overall, 14% of the animals and 44% of the flocks were positive; the mean egg count was 20.5 eggs per gram of faeces. In contrast, no adult paramphistomids were found in roe deer. Older sheep, those from farms located in the central climatic area, without water throughs available on pastures and using their own manure to fertilize, were considered significantly more susceptible to infection with paramphistomids. With respect to egg shedding, animals from 37 to 72 months of age, farms under semiextensive management, and those coinfected with Fasciola hepatica presented significantly greater egg counts. Molecular identification revealed 100% similarity with Calicophoron daubneyi sequences from other European and Mediterranean countries., Conclusions: The present study confirms the increase in the prevalence of paramphistomid infections in sheep in the area with high dissemination of the parasite, as previously reported in cattle, and represents the first molecular identification of C. daubneyi in sheep from Spain. Our results demonstrate that special attention should be given to adult animals since they are the main carriers and are responsible for environmental contamination. In addition, detecting risk areas and applying effective control management measures such as the installation of watering points on pastures seems essential for limiting infections in livestock, especially in sheep, since they are susceptible to developing clinical paramphistomidosis at any time in their lives. The absence of adult flukes in roe deer suggests that they represent less suitable hosts for this trematode than cattle and sheep, although more robust studies monitoring the situation in sympatric areas with domestic ruminants are needed., Competing Interests: Declarations. Ethics approval and consent to participate: All faecal samples used in this study were collected with the permission of the farm owner. All experimental procedures fully complied with European and Spanish ethics regulations on the protection of animals used for scientific purposes (European Directive 2010/63/EU and Spanish Royal Decree 53/2013) and approved by the ethical committee of the University of Santiago de Compostela. Consent for publication: Not applicable. Competing interests: The authors declare that they have no competing interests., (© 2024. The Author(s).)

Background: Several species belonging to the genus Ehrlichia are considered pathogenic for animals and humans. Although wildlife are known to play an important role in the epidemiology of these bacteria, information on the role of wild lagomorphs in their sylvatic cycle is limited. Thus, the objective of the present study was to assess the occurrence of Ehrlichia spp. in ticks collected from wild lagomorphs in Spanish Mediterranean ecosystems., Methods: A total of 1122 pooled ticks (254 pools) collected from 506 wild rabbits (Oryctolagus cuniculus) and 29 Iberian hares (Lepus granatensis) were analysed using a nested PCR assay targeting the partial groEL gene. Ehrlichia spp.-positive samples were further subjected to a second PCR assay targeting 16S rRNA., Results: Three (1.2%) tick pools comprising Rhipicephalus pusillus collected from nine wild rabbits were positive for Ehrlichia spp. All the Ehrlichia DNA sequences were identical, and use of sequence and phylogenetic analyses allowed us to identify a novel Ehrlichia species., Conclusions: We provide evidence that a novel Ehrlichia species, named herein as 'Candidatus Ehrlichia andalusi', which may be of concern for animal and public health, is circulating in R. pusillus in Spanish Mediterranean ecosystems. Further studies are warranted to assess the epidemiology, pathogenicity and zoonotic potential of this Ehrlichia species., (© 2022. The Author(s).)

Background: 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 canis., Case Presentation: A six month old female European shorthair cat from north-western Spain died after being hospitalized for two days. This animal was pregnant and showed anorexia, lethargy, weakness, jaundice and fever with increased respiratory and heart rates. Haematological analysis revealed haemolytic regenerative anaemia, thrombocytopenia and leukocytosis. The presence of piroplasms was assessed using a PCR targeting the 18S rRNA gene of Babesia spp. and Theileria spp.; the sample resulted positive and B. canis was identified by DNA sequence analysis. The possible existence of co-infections with other vector-borne pathogens such as Anaplasma spp., Bartonella spp., Borrelia burgdorferi s.l., Cytauxzoon spp., Ehrlichia spp., Hepatozoon canis, Mycoplasma spp. or Rickettsia spp. was excluded by qPCR., Conclusions: Our results together with previous reports on Babesia infections in cats from Europe suggest that feline babesiosis should be included in the differential diagnosis of animals with anaemia, thrombocytopenia, anorexia and lethargy, especially in young or immunocompromised animals from endemic areas for canine babesiosis., (© 2022. The Author(s).)

Background: Ixodes ricinus, the predominant tick species in Europe, can transmit the causative agents of important human diseases such as Lyme borreliosis (LB), caused by Borrelia spirochetes. In northern Spain, LB is considered endemic; recently, a significant increase of the annual incidence of LB was reported in the northwestern (NW) region., Methods: In order to provide information on the prevalence of Borrelia spp., pooled and individually free-living I. ricinus from NW Spain were molecularly analyzed. Positive samples were characterized at the fla and Glpq genes and the rrfA-rrlB intergenic spacer region to identify Borrelia species/genospecies., Results: Borrelia burgdorferi (sensu lato) (s.l.) individual prevalence and MIR were significantly higher in adult females (32.3 and 16%) than in nymphs (18.8 and 6.2%) and adult males (15.6 and 8.4%). Five Borrelia genospecies belonging to the B. burgdorferi (s.l.) group were identified: B. garinii was predominant, followed by B. valaisiana, B. lusitaniae, B. afzelii and B. burgdorferi (sensu stricto) (s.s.). One species belonging to the tick-borne relapsing fever group (B. miyamotoi) was also found, showing low individual prevalence (1%), positive pool (0.7%) and MIR (0.1%) values. To our knowledge, this is the first citation of B. miyamotoi in free-living ticks from Spain., Conclusions: The significant prevalences of B. burgdorferi (s.l.) genospecies detected in questing ticks from NW Spain are similar to those detected in northern and central European countries and higher to those previously found in Spain. These results together with the high incidence of LB in humans and the high seroprevalence of B. burgdorferi (s.l.) in roe deer shown in other studies reveal that the northwest area is one of the most risky regions for acquiring LB in Spain.