The ascomycete genus Taphrina Fries comprises nearly 100 species recognized by their mycelial states when parasitic on different vascular plants. Whereas the filamentous state is strictly phytoparasitic, the yeast state is saprobic and can be cultured on artificial media. Taphrina species are differentiated mainly on the basis of host range and geographical distribution, type and site of infection and morphology of the sexual stage in infected tissue. However, there has been little progress in the systematics of the genus in recent years, mainly because of the scarcity of molecular studies and available cultures. The main aim of the present study was the reappraisal of species boundaries in Taphrina based on the genetic characterization of cultures (yeast states) that represent about one-third of the currently recognized species. The molecular methods used were (i) PCR fingerprinting using single primers for microsatellite regions and (ii) determination of nucleotide sequences of two approx. 600 bp nuclear rDNA regions, the 59 end of the 26S rRNA gene (D1/D2 domains) and the internal transcribed spacer region (which includes the 5.8S rRNA gene). Sequencing results confirmed the monophyly of the genus (with the probable exclusion of Taphrina vestergrenii) and the combined analysis of the two methods corroborated, in most cases, separation of species defined on the basis of conventional criteria. However, genetic heterogeneity was found within some species and conspecificity was suggested for strains that have been deemed to represent distinct species. Sequences from the ITS region displayed a higher degree of divergence than those of the D1/D2 region between closely related species, but were relatively conserved within species (>99 % identity) and were thus more useful for the effective differentiation of Taphrina species. The results further allowed other topics to be addressed such as the correlation between the molecular phylogenetic clustering of certain species and the respective host plant family and the significance of molecular methods in the accurate diagnosis of the different diseases caused by Taphrina species.

BackgroundCytauxzoonosis is an emerging and life-threatening tick-borne feline disease caused by haemoprotozoan parasites of the genus Cytauxzoon. Information regarding epidemiological and clinical presentation of infections by species other than Cytauxzoon felis is scant. A case of Cytauxzoon sp. infection is described in a 2-year-old mixed breed male domestic cat from Portugal, presenting a history of acute lethargy, anorexia and pyrexia.ResultsComplete blood count revealed a severe anaemia, leucocytosis and thrombocytopenia. A pleural effusion was noticed on thoracic radiograph, and marked splenomegaly and free abdominal fluid were visualized by ultrasound. A molecular screening for the detection of causative agents of infectious anaemia was performed, and a positive result for Piroplasmorida was obtained. DNA sequencing of a 743 bp amplicon of the 18S rRNA gene (GenBank accession no. KU710344) revealed 99.9 % identity with Cytauxzoon manul.ConclusionsThis is the first report of Cytauxzoon sp. (clustering together with C. manul) in a felid from Portugal. Clinical manifestations along with molecular analysis suggest the hypothesis that domestic cats might be infected with and serve as a reservoir host for C. manul.

Anisakiasis is an emerging marine food-borne zoonosis resulting from the accidental ingestion of Anisakis larvae, through the consumption of raw or undercooked infected seafood products. The first case of human gastric hyperinfection by Anisakis simplex with an unusual and severe presentation, occurring in a Portuguese woman, is described in this article. Over 140 anisakid larvae were removed by gastroscopy. Massive infection is uncommon in areas where the consumption of raw fish is not part of the traditional diet, as is the case in Portugal. The increased consumption of raw seafood products is considered a health determinant in the rise in cases of anisakiasis. However, clinicians should be aware of the emergence of these infections, not only because of the new dietary habits of the population, but also because of the high prevalence of Anisakis larvae in the different fish species usually consumed by the population, collected on the Portuguese coast.

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Taphrina Fries is a genus of dimorphic ascomycetes comprising more than 90 species distinguished by the specific infections they produce on different vascular plants. Their filamentous states are restricted to parasitised plant tissue whereas the yeast states are saprobic and can be grown on artificial media. The latter coincide with the anamorphic phases and have been given separate nomenclatural status by the erection of the genus Lalaria R.T. Moore. In its original circumscription Lalaria included only 23 yeast states of known species of Taphrina and its creation was then redundant. Here we describe five novel species in the genus Lalaria to accommodate a total of 44 yeast isolates obtained mainly from leaf surfaces (phylloplane) of different plants in Portugal: Lalaria arrabidae sp. nov. (one strain), L. carpini sp. nov. (one strain), L. inositophila sp. nov. (37 strains), L. kurtzmanii sp. nov. (one strain) and L. veronaerambellii sp. nov. (four strains). L. inositophila was notable for its widespread occurrence since it was recovered during two consecutive years from the leaves of miscellaneous plant species. In the absence of sexual states and of unequivocal associations to particular host plants, the taxonomic relationship of the novel species to the yeast states of Taphrina available from culture collections was verified by the comparative analysis of physiological and molecular characteristics. The latter included PCR fingerprinting using single primers for microsatellite regions, sequencing of the 5' end of the 26S rRNA (LSU) gene (D1/D2 domains) and of the ITS1 and ITS2 rDNA spacer regions, and DNA-DNA hybridisation experiments. An emended description of the genus Lalaria is provided.