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.

Spermatological Characterization of the Cestode Meggittina gerbilli (Cyclophyllidea: Catenotaeniidae), a Parasite of Gerbils, Gerbillus gerbillus and Gerbillus campestris (Rodentia: Muridae) in Tunisia.

Ultrastructural characters of spermiogenesis and the mature spermatozoon of the cestode Meggittina gerbilli (Cyclophyllidea: Catenotaeniidae), a parasite of the Lesser Egyptian gerbil (Gerbillus gerbillus) and the North African gerbil (Gerbillus campestris) (Rodentia: Muridae) in the Djebel Dahar (South of Tunisia), were studied using transmission electron microscopy. The spermiogenesis of M. gerbilli is of Bâ and Marchand's type III, which is mainly characterized by a proximodistal fusion of a single flagellum with a cytoplasmic extension. In this catenotaeniid, the proximal fusion is preceded by a 90° rotation of the flagellum. The spermatozoon is a Levron et al. type VI, which presents a single axoneme with the 9 + '1' trepaxonematan pattern, a periaxonemal sheath, two crest-like bodies, twisted cortical microtubules, and a spiraled nucleus. The obtained results show similarities with the remaining studied catenotaeniids, namely Catenotaenia pusilla and Skrjabinotaenia lobata. The results are compared and discussed according to several characteristics found in the catenotaeniids and other studied cyclophyllideans.

Morphologic and genetic characterization of Pterygodermatites (Mesopectines) valladaresi n. sp. (Nematoda, Rictulariidae), a parasite of the mouse Mus musculus (Rodentia, Muridae) from the Canary Islands (Spain).

A new rictulariid nematode Pterygodermatites (Mesopectines) valladaresi n. sp., parasite of the house mouse Mus musculus (Rodentia: Muridae) in the Canary Islands (Spain) is described by means of light and scanning electron microscopy. The new species belongs to the subgenus Mesopectines characterized by a more or less dorsal orientation of the buccal capsule, the presence of three oesophageal teeth, the morphology of the oral denticles and the Spirurida type of arrangement of caudal papillae in males. The most discriminant characteristics between the new species and the existing species in the subgenus Mesopectines are (a) the number of cuticular projection pairs (62-64), (b) the size of right and left spicules (respectively, 62-90 µm and 123-139 µm), (c) the number of midventral fans in males (3-4), (d) the number of prevulvar/total cuticular projection pairs (38-42/63-71), (e) the posterior differentiation of combs into spines in relation to the position of the vulva and (f) the anterior position of the vulva in relation to the oesophagus-intestine junction in females. Parasitized hosts and geographical distribution are also useful criteria to distinguish P. (Me.) valladaresi n. sp. from the remaining species of the subgenus. In addition, the cox1 sequence of the new species is provided and compared with available data of related species.

Title: Caractérisation morphologique et génétique de Pterygodermatites (Mesopectines) valladaresi n. sp. (Nematoda, Rictulariidae), un parasite de la souris Mus musculus (Rodentia, Muridae) des îles Canaries (Espagne). Abstract: Un nouveau nématode rictulaire, Pterygodermatites (Mesopectines) valladaresi n. sp., parasite de la souris domestique Mus musculus (Rodentia, Muridae) aux îles Canaries (Espagne) est décrit par microscopie optique et électronique à balayage. La nouvelle espèce appartient au sous-genre Mesopectines caractérisé par une orientation plus ou moins dorsale de la capsule buccale, la présence de trois dents œsophagiennes, la morphologie des denticules buccaux et le type Spirurida d'arrangement des papilles caudales chez les mâles. Les caractéristiques les plus discriminantes entre la nouvelle espèce et les espèces existantes du sous-genre Mesopectines sont (a) le nombre de paires de projections cuticulaires (62­64), (b) la taille des spicules droit et gauche (respectivement, 62­90 µm et 123­139 µm), (c) le nombre de bourrelets cuticulaires médio-ventraux chez les mâles (3­4), (d) le nombre de peignes prévulvaires par rapport au nombre total de peignes (38­42/63­71), (e) la transformation en épines des peignes après la vulve et (f) la position antérieure de la vulve par rapport à la jonction œsophage-intestin chez les femelles. Les hôtes parasités et la répartition géographique sont également des critères utiles pour distinguer P. (Me.) valladaresi n. sp. des autres espèces du sous-genre. De plus, la séquence du cox1 de la nouvelle espèce est fournie et comparée aux données disponibles sur les espèces apparentées.

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.

New Data on Pterygodermatites (Pterygodermatites) plagiostoma Wedl, 1861 (Nematoda, Rictulariidae) Parasite of the Algerian Hedgehog Atelerix algirus Linnaeus, 1758 (Eulipotyphla: Erinaceidae) from the Canary Islands.

A redescription of the rictulariid nematode Pterygodermatites (Pterygodermatites) plagiostoma Wedl, 1861, is made by means of light and scanning electron microscopy, including the first data on male specimens. The morphologic study was based on specimens recovered from two Algerian hedgehogs (Atelerix algirus) from Tenerife and Gran Canaria islands (Canary Archipelago, Spain). The main characteristics of P. (P.) plagiostoma males are the presence of 49-53 pairs of cuticular projections, the presence of one or two midventral precloacal fans (generally one), and the size of two unequal spicules, measuring 98-123 µm (right spicule) and 185-236 µm (left spicule). The cloacal papillae are arranged according to the Ascaridida type. They include two precloacal pairs, an unpaired precloacal papilla, one pair lateral to the cloaca, six postcloacal pairs, and a pair of phasmids near the tail tip. Females are mainly characterized by the presence of 71-77 pairs of cuticular projections, with 43-46 pairs of prevulvar combs, by the differentiation from combs to spines at the level of or slightly posterior to the vulva and by the position of the vulva, located posteriorly to the esophagus-intestine junction. Clear differences were found between P. (P.) plagiostoma and related species of the subgenus P. (Pterygodermatites).

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.

First Coronavirus Active Survey in Rodents From the Canary Islands.

Since the beginning of the 21st century five new coronaviruses inducing respiratory diseases in humans have been reported. These emergences has promoted research on coronaviruses in wildlife. We started the first eco-epidemiological study to screen the presence of coronaviruses circulating in mice and rats of four Canary Islands. Between 2015 and 2019, we obtained fecal samples of three rodent species (150 Mus musculus, 109 Rattus rattus and 1 Rattus norvegicus) captured in urban and rural areas. Fecal samples were analyzed by nRT-PCR and the resulting sequences were compared to known diversity using Bayesian phylogenetic methods. We only found coronavirus RNA in house mice from El Hierro (10.53%), Tenerife (7.02%) and Lanzarote (5.26%) islands. All coronaviruses detected belong to the species Murine coronavirus belonging to the genus Betacoronavirus and subgenus Embecovirus, being all positive house mice captured in anthropogenic environment. The phylogenetic analysis shows that murine coronaviruses from the Canary Islands are related to European murine coronaviruses. Albeit data are still scarce in the region, the most probable origin of M. coronavirus present in the Canary Islands is continental Europe. According to temporal Bayesian phylogenetics, the differentiation between Canary and continental viruses seems to be quite recent. Moreover, murine coronaviruses from El Hierro, Tenerife and Lanzarote islands tend to segregate in different clades. This enlightens the potential role of rodents or other possibly invasive species in disseminating infectious diseases to remote places through exchanges with the continent. It is important to consider these aspects in the sanitary control of islands, for health and biodiversity preservation concerns.

A Peculiar Distribution of the Emerging Nematode Angiostrongylus cantonensis in the Canary Islands (Spain): Recent Introduction or Isolation Effect?

Angiostrongylus cantonensis is an emerging zoonotic nematode recognized as the leading cause of eosinophilic meningitis in the word. After its discovery in China, it was recorded in 30 countries worldwide. Recently, it has expanded to new areas such as South America and it has been recently found in the Atlantic island of Tenerife (Canary Islands). In order to characterize the distribution of A. cantonensis in the Canary Islands, the lungs of 1462 rodents were sampled in eight islands of the archipelago over 13 years and were then analyzed for A. cantonensis. Remarkably, the parasite was detected only in Tenerife, in Rattus rattus (19.7%) and Rattus norvegicus (7.14%). They were concretely in the northern part of the island, which had a warmer and more humid climate than the south and main cities. The absence of this nematode in other islands with similar environmental conditions could be explained by an isolation effect or by a recent introduction of the parasite in the islands. Besides, the presence in Tenerife of the most invasive lineage of A. cantonensis reinforced the hypothesis of a recent introduction on this island. This study highlights the need to implement control measures to prevent the expansion to other areas in order to avoid the transmission to humans and other animals.

Study of Zoonotic Enteric Pathogens of Atelerix algirus in Tenerife, Canary Islands, Spain.

Atelerix algirus is an invasive species in the Canary Islands (Spain). There are few studies about the zoonotic pathogens this species could be hosting; therefore, this study was focused on analyzing causative agents of diarrhea in humans in feces from hedgehogs. A total of 45 fecal samples obtained in Tenerife (Canary Islands) were analyzed in this study using Biofire FilmArray gastrointestinal panel with an integrated Biofire FilmArray system. Forty-two (93.33%) of the samples presented at least one of the pathogens detected by the panel. The prevalence of four bacteria stands out as for enteropathogenic Escherichia coli (71.11%), Salmonella (66.67%), Clostridioides difficile (33.33%), and Campylobacter sp. (22.22%), all of which were widely distributed along Tenerife. Besides, other pathogens were found, Cryptosporidium sp. and enterotoxigenic E. coli lt/st in 6.66% of the animals, Shigella/enteroinvasive E. coli in 4.44%, and Norovirus GI/GII, Plesiomonas shigelloides, and Vibrio sp. in 2.22%. Of the hedgehogs, 26.66% were hosting just one pathogen, and the others showed coinfection: 24.44% hosted two, 31.11% hosted three, and 11.11% hosted four or more. The close contact with hedgehogs may imply the transmission of not only one causative agent of diarrhea but also multiple agents, since coinfection is highly prevalent. The lack of management measurements for this animal in the Canary Islands, the common habit of adopting hedgehogs from wildlife without veterinary control, and the fact that most of the hedgehogs studied belonged to highly populated areas imply a high risk of transmission of pathogens to humans.

Bartonella species and haplotypes in rodents and their fleas in Lanzarote and El Hierro in the Canary Islands, Spain.

Because isolated ecosystems contribute to species variability, especially oceanic island ecosystems, the present work focused on the study of the Bartonella species and haplotypes in Lanzarote and El Hierro, two Canary islands with evident bioclimatic differences between them. A total of 123 rodents and 110 fleas from two islands were screened for the presence of Bartonella by PCR analysis of the gltA and nuoG genes. The overall prevalence was 5.7% in rodents and 20.4% in fleas. A total of seven gltA-haplotypes was found in both rodents and fleas, belonging to the species Bartonella mastomydis and Bartonella tribocorum in Lanzarote, and to Bartonella rochalimae and Bartonella elizabethae in El Hierro, as well as recently described species Bartonella kosoyi in both islands. Besides, potential co-infections were detected based on the nuoG analysis. Further, Xenopsylla cheopis was the only flea species identified. Our study shows that isolated ecosystems such as the Canary Islands lead to the appearance of new Bartonella haplotypes along different biotopes, with diverse flea species involved in the spreading of the pathogen being of great relevance due to the zoonotic potential of the species found.

Pathogenic Leptospira species in rodents from Corsica (France).

Leptospirosis is a worldwide emerging zoonotic disease caused by Leptospira species, that in some patients develop severe forms with high mortality. In France, Corsica is the area where the highest incidences have been reported. The present study was focused on the analysis of pathogenic Leptospira species in rodents of Corsica, as these micromammals are the main natural reservoirs of the bacteria, in order to identify the circulating species and to locate possible risk focuses of transmission, as no previous study on the presence of Leptospira species has been carried out in the island. Rattus rattus, Rattus norvegicus, Apodemus sylvaticus and Mus musculus domesticus were captured in the proximity of water sources along Corsica, the detection of pathogenic Leptospira species was carried out by amplification of the LipL32 gene. The bacteria were found in all the rodent species analyzed and widely. The general prevalence was 10.4%, reaching the maximum value in Bastia (45%). Leptospira interrogans and Leptospira borgpetersenii were identified by phylogenetic analysis, but also two sequences which corresponded to an unnamed Leptospira species, only previously found in rodents of New Caledonia. The high incidence of human leptospirosis in Corsica could be partially explained by the wide distribution of pathogenic Leptospira species identified in this study. Also, the presence of an unknown pathogenic species of Leptospira in an area with high prevalence, may be involved in the higher incidence of Leptospirosis in this island, however, the zoonotic capacity of this species remains unknown. The results obtained are interesting for public health since all positive samples were found near water sources and one of the routes of transmission of leptospirosis is contact with contaminated water. This information could help the competent entities to take preventive measures, reducing the incidence of human leptospirosis in Corsica.

First ocular report of Gurltia paralysans (Wolffhügel, 1933) in cat.

A two-year-old cat from Tenerife, Canary Islands (Spain), presented with two months history of uveitis in the left eye. It had been treated for two months but still showed an active uveitis. After new treatment, eye examination showed a mobile worm in the anterior chamber. Following surgical removal, the worm was obtained. Morphological study revealed that it was a male metastrongyloid nematode (Nematoda: Metastrongyloidea) with caudal bursa and two similar spicules. Molecular tools based on the 18Sr RNA gene sequence identified the parasite as Gurltia paralysans, which is a neurotropic nematode previously found in South America. Therefore, this article describes the first report of ophthalmic case of parasitism by G. paralysans, and cites for first time this species out of South America.

Bartonella in Rodents and Ectoparasites in the Canary Islands, Spain: New Insights into Host-Vector-Pathogen Relationships.

Bartonella genus is comprised of several species of zoonotic relevance and rodents are reservoirs for some of these Bartonella species. As there were no data about the range of Bartonella species circulating among rodents in the Canary Islands, our main aim was to overcome this lack of knowledge by targeting both the citrate synthase (gltA) and the RNA polymerase beta subunit (rpoB) genes. A total of 181 small mammals and 154 ectoparasites were obtained in three of the Canary Islands, namely Tenerife, La Palma, and Lanzarote. The overall prevalence of Bartonella DNA in rodents was 18.8%, whereas the prevalence in ectoparasites was 13.6%. Bartonella sequences closely related to the zoonotic species Bartonella elizabethae, Bartonella tribocorum, and Bartonella rochalimae were identified in rodents, whereas two different gltA haplotypes similar to B. elizabethae were also detected in fleas. Furthermore, Bartonella queenslandensis DNA was also identified in rodents. A strong host specificity was observed, since B. elizabethae DNA was only found in Mus musculus domesticus, whereas gltA and rpoB sequences closely related to the rest of Bartonella species were only identified in Rattus rattus, which is probably due to the host specificity of the arthropod species that act as vectors in these islands. Our results indicate that humans may contract Bartonella infection by contact with rodents in the Canary Islands.

Emerging rabbit haemorrhagic disease virus 2 (RHDV2) at the gates of the African continent.

Until the beginning of this decade, the genetic characterization of rabbit haemorrhagic disease virus (RHDV) from Iberian Peninsula had revealed the existence of two genogroups, G1 and sporadically G6. In 2010, the new emerging rabbit haemorrhagic disease variant, RHDV2 or RHDVb, was described in France, from where it has rapidly spread throughout Europe, including Iberian Peninsula countries. Nevertheless, although cases of rabbit haemorrhagic disease (RHD) have been reported in the Canary Islands, a Spanish archipelago located 100km off the coast of Morocco, no genetic characterization of RHDV had been carried out. Consequently, in order to identify the circulating RHDV strains in this archipelago, liver samples of six farm rabbits and fifteen wild rabbits were collected from several areas of the largest island, Tenerife, and analyzed for the presence of RHDV by antigen capture double antibody sandwich ELISA. In case of positive ELISA result, we amplified and sequenced two fragments of the vp60 gene, which were concatenated for phylogenetic purposes. The sequences analysis revealed the presence of RHDV2 in both farm and wild rabbits from several areas of Tenerife. This result constitutes the first finding of RHDV2 in the Canary Islands. These RHDV2 strains found in Tenerife shared two exclusive SNPs that have not been observed in the rest of RHDV2 strains. The identification of RHDV2 and the absence of classic RHDV strains in this study suggest that RHDV2 may be replacing classic strains in Tenerife, as has been also proposed in Iberian Peninsula, France and Azores. Given the proximity of the Canary Islands to the African continent, this result should raise awareness about a possible dispersal of RHDV2 from the Canary Islands to the North of Africa.