Mitochondrial sequences of Rhipicephalus and Coxiella endosymbiont reveal evidence of lineages co-cladogenesis.

Rhipicephalus ticks are competent vectors of several pathogens, such as Spotted Fever Group Rickettsiae (SFGR) and many Babesia species. Within this genus, different R. sanguineus s.l. lineages show an unequal vector competence and resistance regarding some pathogenic strains. Current literature supports that tick endosymbionts may play an essential role in the transmission ability of a vector. Indeed, the microbial community of Rhipicephalus seems to be dominated by Coxiella-like endosymbionts (CLE). Still, their co-evolutionary associations with the complicated phylogeny of Rhipicephalus lineages and their transmissible pathogens remain unclear. We performed a phylogenetic congruence analysis to address whether divergent R. sanguineus s.l. lineages had a different symbiont composition. For that, we applied a PCR based approach to screen part of the microbial community present in 279 Rhipicephalus ticks from the Iberian Peninsula and Africa. Our analyses detected several qPCR-positive signals for both SFGR and Babesia species, of which we suggest R. sanguineus-tropical lineage as a natural vector of Babesia vogeli and R. sanguineus-temperate lineage of SFGR. The acquisition of 190 CLE sequences allowed to evaluate co-phylogenetic associations between the tick and the symbiont. With this data, we observed a strong but incomplete co-cladogenesis between CLE strains and their Rhipicephalus tick lineages hosts.

Plastic ingestion in aquatic birds in Portugal.

In modern society, plastic items have become indispensable. The rapid growth of plastic production has led to an increase in the concentration of plastic waste in the environment and, consequently, wildlife has been severely affected. As wide-ranging foragers and predators, aquatic birds are ideal sentinels for monitoring changes in their environment. Plastic found in stomach contents of stranded aquatic birds collected throughout Portugal was examined. Out of the 288 birds processed, 12.9% ingested plastics. Six of the 16 species assessed showed evidence of plastic ingestion. The Lesser Black-backed Gull (18.7%) had the highest incidence while, among those that did ingest plastics, the Northern Gannet (4.8%) had the lowest. User plastics were the most common type of plastic ingested, while microplastics and off/white-clear were the most common size and colour respectively of plastics found. This study sets a first multispecies baseline for incidence of plastic ingestion by aquatic birds in Portugal.

Plastic ingestion in aquatic-associated bird species in southern Portugal.

Excessive use of plastics in daily life and the inappropriate disposal of plastic products are severely affecting wildlife species in both coastal and aquatic environments. Birds are top-predators, exposed to all threats affecting their environments, making them ideal sentinel organisms for monitoring ecosystems change. We set a baseline assessment of the prevalence of marine plastic litter affecting multi-species populations of aquatic birds in southern Portugal. By examining 160 stomach contents from 8 species of aquatic birds, we show that 22.5% were affected by plastic debris. Plastic was found in Ciconia ciconia, Larus fuscus and L. michahellis. Ciconia ciconia ingested the highest amount (number of items and total mass) of plastic debris. Polydimethylsiloxane (PDMS, silicones) was the most abundant polymer and was recorded only in C. ciconia. Plastic ingestion baseline data are of crucial importance to evaluate changes through time and among regions and to define management and conservation strategies.

Tick-borne bacteria and protozoa detected in ticks collected from domestic animals and wildlife in central and southern Portugal.

Ticks are vectors of many human and animal pathogens. The aim of this study was to screen bacteria and protozoa from ticks infesting domestic animals and wildlife collected in central and southern Portugal. A total of 593 ticks, comprising 465 (78.4%) adults, 122 (20.6%) nymphs, and six (1.0%) larvae, were collected from 283 hosts of 25 different species (4 domestic and 21 wild). Overall, the analysis of DNA extracts prepared from ticks collected from hosts of 11 different species in the districts of Castelo Branco, Portalegre, Lisboa, Setúbal, Beja and Faro, revealed the presence of genomic sequences from Anaplasma sp., A. ovis, Babesia sp., relapsing fever-like Borrelia sp., Ehrlichia spp., Rickettsia aeschlimannii, Ri. helvetica, Ri. massiliae, Ri. raoultii, Ri. slovaca, Candidatus Ri. barbariae, Theileria annulata and T. ovis, in specimens of Dermacentor marginatus, Hyalomma lusitanicum, Hy. marginatum, Rhipicephalus bursa and Rh. sanguineus sensu lato. The obtained results suggest the circulation of a wide variety of infectious agents, some of zoonotic concern, in hard ticks from Portugal. Further studies should be conducted to better characterize (both genetically and phenotypically) the putative novel microorganisms detected, both in what regards their potential pathogenity towards vertebrates, and to assist the implementation of effective control strategies for the management of ticks and human and animal tick-borne pathogens.

An outbreak of philophthalmosis in Larus michahellis and Larus fuscus gulls in Iberian Peninsula.

Trematodes of the genus Philophthalmus Loos, 1899 are the eye parasites of birds and mammals, which use freshwater snails as their first intermediate hosts. Here we examined the presence of philophthalmids in a total of 1515 gulls (589 Larus fuscus and 926 Larus michahellis) admitted between January 2010 and October 2016 for rehabilitation at Olhão (Portugal), by the use of combined morphological and molecular analysis. We recorded the first infected L. fuscus and L. michahellis in July and November 2015, respectively. The philophthalmids were located in the conjunctival sac or under the nictitating membrane. Gulls infected with Philophthalmus lucipetus Rudolphi, 1819 presented no clinical signs, while those infected with Philophthalmus lacrymosus Braun, 1902 presented serious eye damage in the same host species. The prevalence of P. lucipetus reached 3.6% in L. fuscus and 0.8% in L. michahellis; the prevalence of P. lacrymosus reached 0.3% and 0.0%, respectively. The outbreak of P. lucipetus likely started in a narrowly defined area, since the first six cases, found between July and October 2015, originated from a single municipality, and only later more cases started to be retrieved from other municipalities of Portugal. These findings represent the first records of both philophthalmids in the Iberian Peninsula, their first records in L. michahellis and the first record of P. lacrymosus in L. fuscus. Further follow-up of the outbreak and the identification of intermediate hosts are needed.

Extension of the Avian Host Range of Collyriclosis in Europe.

We describe cases of collyriclosis in apodiform and passeriform birds in Portugal, Switzerland, and Germany. We extend the host range of Collyriculm faba to include apodiform birds ( Apus apus , Apus melba , and Apus pallidus ) and the passerine Sitta europaea (Eurasian Nuthatch). Infections varied in severity from an incidental finding to severe debilitation and death. The infection route remains unclear with the apparent absence from Germany, Portugal, and Switzerland of the first intermediate host of C. faba, the aquatic gastropod Bythinella austriaca, implying that other organisms might be involved in the parasite's life cycle. Furthermore, the detection of C. faba cysts in very young passerine birds may indicate an infection during the nestling stage and a rapid development of parasite-containing subcutaneous cysts. This series of cases highlights an increased geographic range into Portugal and the potential debilitating nature of a parasite of migratory birds in Europe. However, given the rarity of cases, collyriclosis does not seem to present an important threat to migratory species preservation.

Cercopithifilaria sp. II in Vulpes vulpes: new host affiliation for an enigmatic canine filarioid.

Cercopithifilaria bainae and Cercopithifilaria grassii (Spirurida, Onchocercidae) are filarioids inhabiting the skin of dogs worldwide. The microfilariae of a third species namely, Cercopithifilaria sp. II sensu Otranto et al. 2013, have been morphologically and molecularly characterized but scientific knowledge of this parasite is minimal. The first case of infection of a red fox (Vulpes vulpes) with the filarioid Cercopithifilaria sp. II is herein described in Castro Marim, Portugal. Microfilariae from skin sediment of the fox's ear were morphological characterized, and the identification was confirmed molecularly in samples from skin sediment, skin samples, and from Rhipicephalus sanguineus group ticks collected from the animal (99% homology with Cercopithifilaria sp. II). Studies should evaluate if red foxes might play a role in the maintenance and distribution of Cercopithifilaria sp. II infection in dog populations.