High Prevalence of Microsporidia in the North African Hedgehog (Atelerix algirus) in the Canary Islands, Spain.

Microsporidia are unicellular eukaryotic obligate intracellular parasites with a wide range of hosts reported worldwide; however, little is known about the epidemiological data on microsporidia infection in animals from the Canary Islands. Since data on microsporidia infection in hedgehog species are scarce, the aim of this study was to analyze the presence and identity of microsporidia in a group of North African hedgehogs (Atelerix algirus) using microscopic and molecular methods. From December 2020 to September 2021, a total of 36 fecal samples were collected from naturally deceased hedgehogs from Tenerife and Gran Canaria. All samples showed spore-compatible structures (100%; 36/36) under microscopic analysis, of which 61.1% (22/36) were amplified via the nested-polymerase chain reaction (PCR) targeting the partial sequence of the 16S rRNA gene, the internal transcribed spacer (ITS) region, and the partial sequence of the 5.8S rRNA gene. After Sanger sequencing and ITS analysis, Enterocytozoon bieneusi was detected in 47.2% (17/36) of the samples, identifying two novel genotypes (AAE1 and AAE2), followed by the detection of an undetermined species in 8.3% (3/36) and Encephalitozoon cuniculi genotype I in 5.6% (2/36) of the samples. This study constitutes the first report of microsporidia species in Atelerix algirus worldwide, highlighting the high prevalence of zoonotic species.

Metastrongyloid Infection with Aelurostrongylus abstrusus, Troglostrongylus brevior, Oslerus rostratus and Angiostrongylus chabaudi in Feral Cats from the Canary Islands (Spain).

Lungworms are a major cause of feline respiratory disease, frequently underdiagnosed due to its presentation of symptoms being similar to that of other feline respiratory pathologies. Epidemiological data about these nematodes are scarce in the Canary Islands (Spain). Given the veterinary importance of these parasites, the aim of the present study was to improve the current epidemiological knowledge of the lungworm species that could be affecting feral cats in this archipelago. A total of 29 feral cats from La Gomera were analyzed. The respiratory tract of each animal was inspected and the nematodes obtained were identified by morphological keys and molecular techniques. Metastrongylids were detected to be widely distributed throughout the island with a prevalence of 55.2% (16/29). The species Aelurostrongylus abstrusus, Troglostrongylus brevior, Oslerus rostratus and Angiostrongylus chabaudi were identified. Also, coinfections with A. chabaudi and O. rostratus were detected in two animals. The present study shows a high diversity of lungworms in feral cats in La Gomera, with the first report of A. chabaudi and T. brevior for the Canary Archipelago and the first citation of A. chabaudi in cats for Spain. The wide distribution and high prevalence found in this study indicate a high risk of exposure to pulmonary infections in cats.

Data on New Intermediate and Accidental Hosts Naturally Infected with Angiostrongylus cantonensis in La Gomera and Gran Canaria (Canary Islands, Spain).

Angiostrongylus cantonensis is a metastrongyloid nematode and the etiologic agent of angiostrongyliasis, a disease characterized by eosinophilic meningitis. This emerging zoonotic parasite has undergone great expansion, including in some regions of Europe and America. In the Canary Islands, the parasite was first discovered parasitizing Rattus rattus on the island of Tenerife in 2010. To date, the distribution of this parasite in the Canary Islands has been restricted to the northern zone and the main cities of Tenerife. Using molecular tools for the sentinel species present in the Canary Islands, this study confirmed the presence of the nematode on two other islands in the Canary Archipelago: La Gomera and Gran Canaria. Furthermore, this emerging parasite was detected, besides in the common definitive host R. rattus, in wild Mus musculus and Felis catus and in four terrestrial gastropod species, Limacus flavus, Milax gagates, Insulivitrina emmersoni, and Insulivitrina oromii, two of them endemic to La Gomera, for the first time, increasing the number of non-definitive host species. This study reinforces the expansion character of A. cantonensis and highlights the importance of knowledge about sentinel species for identifying new transmission locations that help prevent and control the transmission of the parasite and, thus, prevent public health problems.

Molecular Detection of Microsporidia in Rabbits (Oryctolagus cuniculus) in Tenerife, Canary Islands, Spain.

Enterocytozoon bieneusi and Encephalitozoon spp. are microsporidia with zoonotic potential that have been identified in humans, as well as in a large group of wild and domestic animals. Several wildlife species have been studied as reservoirs of zoonotic microsporidia in mainland Spain, including the European rabbit (Oryctolagus cuniculus). Due to a lack of data on microsporidia infection in wildlife on the Canary Islands, the aim of this work was to analyze the prevalence and identify the species of microsporidia in rabbits in Tenerife. Between 2015 and 2017, a total of 50 fecal samples were collected from rabbits in eight municipalities of Tenerife, Canary Islands, Spain. Seven of the fifty samples (14%) were amplified using nested polymerase chain reaction (PCR) targeting the partial sequence of the 16S rRNA gene, the internal transcribed spacer (ITS) region, and the partial sequence of the 5.8S rRNA gene. Sanger sequencing reveals the presence of Encephalitozoon cuniculi genotype I in two samples (4%), and undescribed microsporidia species in five samples (10%). This study constitutes the first molecular detection and genotyping of E. cuniculi in rabbits in Spain.

Morphological and Molecular Identification of Anisakis spp. (Nematoda: Anisakidae) in Commercial Fish from the Canary Islands Coast (Spain): Epidemiological Data.

The study aimed to perform the molecular identification of Anisakis larvae in commercial fish from the coast of the Canary Islands and to provide data on their infection level for the host and the species of this nematode parasite that we could find in several species of commercial interest in the Canary Archipelago. Fish specimens (n = 172) from the Canary coasts were examined for parasites. In total, 495 larvae were identified; PCR was carried out for the entire ITS rDNA and cox2 mtDNA region, obtaining sixteen sequences for the entire ITS rDNA region and fifteen for the cox2 mtDNA, this being the first contribution of nucleotide sequences of Anisakis species of fish caught from the Canary Islands. An overall prevalence of 25% was obtained in the fish analyzed, and five species of Anisakis were identified, these being Anisakis simplex (s.s.), Anisakis pegreffii, Anisakis physeteris, Anisakis nascettii and Anisakis typica and the hybrid Anisakis simplex x Anisakis pegreffii. The results obtained in this study have relevance for public health, since the pathology will depend on the species of Anisakis, so it is important to know the health status of fish in the waters of the Canary Islands to assure a safer consumption and take adequate measures, in addition to the provision of epidemiological data.

Ultrastructural and molecular study of the microsporidian Toguebayea baccigeri n. gen., n. sp., a hyperparasite of the digenean trematode Bacciger israelensis (Faustulidae), a parasite of Boops boops (Teleostei, Sparidae).

A new microsporidian Toguebayea baccigeri n. gen., n. sp., hyperparasite of Bacciger israelensis (Digenea, Faustulidae), parasite of Boops boops (Teleostei, Sparidae) is described by means of transmission electron microscopy. The phylogenetic analysis, based on the SSU rDNA gene, places the new species in the clade containing mainly crustacean-infecting microsporidia of the genus Cucumispora, within superclade V (Marinosporidia) sensu Vossbrinck et al., 2014. Mature spores of T. baccigeri are ovoid, uninucleated and measure 2.5 × 1.4 µm. The number of coils of the polar tube is 8-10. The polaroplast is composed of an external lamellar part and an internal vesicular or granular part. The main differences that distinguish the new genus and new species from the closely related microsporidia include hyperparasitism in a digenean host infecting a marine fish, the geographic distribution (coast of Tunisia), presence of one nucleus at all developmental stages, disporoblastic sporogony, and the absence of sporophorous vacuoles.

TITLE: Étude ultrastructurale et moléculaire de la microsporidie Toguebayea baccigeri n. gen., n. sp., hyperparasite du trématode digène Bacciger israelensis (Faustulidae), parasite de Boops boops (Teleostei, Sparidae). ABSTRACT: Une nouvelle microsporidie Toguebayea baccigeri n. gen., n. sp., hyperparasite de Bacciger israelensis (Digenea, Faustulidae), parasite de Boops boops (Teleostei, Sparidae) est décrite en microscopie électronique à transmission. L'analyse phylogénétique, basée sur l'étude du gène SSU rDNA, place la nouvelle espèce dans le clade qui contient notamment les microsporidies parasites de crustacés du genre Cucumispora, dans le superclade V (Marinosporidia) sensu Vossbrinck et al, 2014. Les spores mûres de T. baccigeri sont ovoïdes, uninucléées et mesurent 2,5 × 1,4 µm. Le nombre de tours de spire décrits par le tube polaire est de 8 à 10. Le polaroplaste est composé d'une partie externe lamellaire et d'une partie interne vésiculeuse ou granuleuse. Les principales différences entre le nouveau genre et la nouvelle espèce et les microsporidies les plus proches sont : l'hyperparasitisme d'un digène parasite d'un poisson marin, la localisation géographique (côtes de la Tunisie), et certains aspects du développement, notamment la présence d'un seul noyau à tous les stades de développement, la sporogonie disporoblastique et l'absence de vacuoles sporophores.

Molecular Detection of Cryptosporidium cuniculus in Rabbits (Oryctolagus cuniculus) from Tenerife, Canary Islands, Spain.

Cryptosporidium cuniculus is a zoonotic parasite responsible for cryptosporidiosis cases and outbreaks in both humans and rabbits. Since there are no molecular Cryptosporidium spp. infection data in rabbits (Oryctolagus cuniculus) from Spain, our aim was to gather information about this parasite in wild European rabbits from Tenerife, Canary Islands (Spain). A total of 100 faecal samples were collected from rabbits from eight municipalities of Tenerife. Microscopic analysis showed that 4.0% of the samples presented structures compatible with Cryptosporidium oocyst. A nested polymerase chain reaction (PCR) targeting 18S ribosomal RNA (rRNA) gene fragments was carried out, and sequencing confirmed the identity of C. cuniculus in one sample (1.0%). The sample was successfully subtyped using nested PCR analysis of the 60-kDa glycoprotein (gp60) gene as the subtype VbA26R3. This study confirms the presence of C. cuniculus in wild rabbits from Tenerife, providing new information on the occurrence of this zoonotic parasite. Further studies are required to better understand the epidemiology of Cryptosporidium spp. in wild rabbits in Spain and their possible public health repercussions.