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.

Study of tick-borne zoonotic pathogens in questing and feeding ticks in Tenerife, Canary Islands, Spain.

Ticks are vectors of many zoonotic pathogens of clinical relevance, including Anaplasma and Rickettsia species. Since few data about these tick-borne pathogens are available in the Canary Islands, the aim of the present study was to screen their presence in questing and feeding ticks on the island of Tenerife. A total of 81 ticks was removed from six hedgehogs, and eight ticks were collected from the grass. PCR assays for tick species identification based on the LSU rRNA gene, and detection of Anaplasma spp. and Rickettsia spp. by targeting the 16s rRNA and gltA gene, respectively, were carried out. Rhipicephalus sanguineus and Haemaphysalis formosensis tick species were identified, with Rickettsia hoogstraalii detected in H. formosensis. In addition, Rickettsia helvetica and one unidentified Rickettsia species were detected. The overall prevalence of Rickettsia in ticks was 12.2%, and none harbored Anaplasma sp. This work constitutes the first identification in the Canary Islands of pathogenic R. helvetica and the species of unknown pathogenicity R. hoogstraalii in questing ticks. The clinical relevance of the pathogens identified highlights the need for studies with increased sample size and locations, including potential hosts, as well as warning the population about the relevance of ticks as vectors.

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.

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.

Molecular detection of Bartonella henselae, Bartonella clarridgeiae and Rickettsia felis in cat and dog fleas in Tenerife, Canary Islands, Spain.

The cat flea Ctenocephalides felis is the main vector of Bartonella henselae and Bartonella clarridgeiae, the causative agents of cat-scratch disease (CSD) and the spotted-fever agent Rickettsia felis. In spite of their worldwide distribution, there are no data on the occurrence of CSD-causing Bartonella species or the prevalence of Rickettsia species in the Canary Islands, Spain. Therefore, the aim of our study was to screen cat and dog fleas for both pathogens. A total of 128 C. felis from cats and dogs were screened for Bartonella and Rickettsia by PCR. Bartonella henselae (2.3%) and B. clarridgeiae (3.9%) were found in fleas infesting cats, whereas R. felis was identified in both cat (36.6%) and dog (40.7%) fleas. Further, co-infections were observed. This work constitutes the first finding of CSD-causing Bartonella species and the first study on the prevalence of R. felis in fleas from domestic animals in the Canary Islands. These results indicate public health importance, as associated infections could be misdiagnosed in the Archipelago despite their clinical relevance. Establishing human and animal routine diagnosis procedures for these pathogens along with improving vector control in shelters is necessary in order to prevent the spread of the infections among animals.

Evaluating Different Methodologies for Bioprospecting Actinomycetes in Canary Islands Soils.

Actinomycetes are a wide group of Gram positive prokaryotes, the soil being their most characteristic habitat, where they play important ecological functions. Their immense biotechnological potential as producers of bioactive molecules of great commercial and industrial interest is exemplified by most antibiotics for clinical use being derived from this group of bacteria. In this work several methodologies and culture media were tested for the isolation of actinomycetes in soils from three different edafoclimatic areas of Tenerife (Canary Islands, Spain): an arid zone of the southeast coast of the island, a humid area in the laurel forest and the Canary high mountain. The results of this work evidenced that just an air drying of the sample during 7 days (pretreatment) produces high counts of actinomycetes versus non-actinomycetes in the three soils and in any of the culture media studied, making unnecessary post physical and chemical treatments. However, the only method that produced an exclusive isolation of actinomycetes was the use of 0.22-µm filters as a physical barrier, and hence being a tool of unquestionable utility to estimate actinomycetes populations in soil.The analysis by BOX rep-PCR determined that the richness and genetic diversity of the isolates of these localities are very high. The greatest similarities were, with one exception, found between strains from the same locations. Izaña was one that obtained the highest diversity. Regarding the biotecnology potential as antibiotic produces isolates from Güimar soils are the most promising as 7 out of 10 isolates showed some antibiotic activity.

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.

Vertebrate Ticks Distribution and Their Role as Vectors in Relation to Road Edges and Underpasses.

Roads fragment vertebrate populations and affect the dynamics and dispersal patterns of vertebrate parasites. We evaluated how vertebrate ticks distribute near roads and road underpasses in human-caused road-fragmented landscapes in Seville, SW Spain. We sampled 49 stations with 93 individualized sampling points and assessed tick abundance. We explored the relationship between tick presence and abundance and distance to the nearest road and underpass (drainage culverts and other passages used by vertebrates), and landscape features through categorical regression and nonparametric statistics. The presence of the tick-borne pathogens Borrelia sp. and Bartonella sp. was also analyzed by PCR. We found preliminary evidence of high relative tick abundance next to roads and in the vicinity of road underpasses. Plant cover type was related to tick presence in this road context. Implications of road permeability and edge effect in patterns of vertebrate-tick relationships in road fragmentation contexts are discussed. Both Borrelia sp. and Bartonella sp. were detected in the ticks analyzed. This is the first report of these bacteria in ticks from Seville. The results confirm the potential risk of acquiring Lyme disease and bartonellosis in this area.

Bartonella spp. in Small Mammals, Benin.

This study aimed to investigate the prevalence and genetic diversity of Bartonella organisms in small mammals in Cotonou, Benin. We captured 163 rodents and 12 insectivores and successfully detected Bartonella DNA from 13 of the 175 small mammal individuals. Bartonella spp., identical or closely related to Bartonella elizabethae, Bartonella tribocorum, and Bartonella rochalimae, was detected. A potential new Bartonella species, proposed as Candidatus Bartonella mastomydis, was found in three Mastomys individuals and genetically characterized by targeting two housekeeping genes (rpoB and gltA) and the intergenic species region. However, 20.8% of gray rats were found to be infected with Bartonella spp., and none of the black rats analyzed was positive. This work may be important from a public health point of view due to the zoonotic nature of the Bartonella species detected and warrants further investigation on the unknown zoonotic potential of this newly proposed Bartonella species.

Intermediate hosts of Angiostrongylus cantonensis in Tenerife, Spain.

The nematode Angiostrongylus cantonensis is the causative agent of human angiostrongyliasis, the main clinical manifestation of which is eosinophilic meningitis. Although this parasite has been found recently in its definitive rat host in Tenerife (Canary Islands, Spain), showing a widespread distribution over the north-east part of the island, there are no available data regarding which snail and/or slug species are acting as intermediate hosts on this island. Consequently, the objective of this work was to determine the possible role of three mollusc species, Plutonia lamarckii, Cornu aspersum and Theba pisana, as intermediate hosts of A. cantonensis in Tenerife. Between 2011 and 2014, 233 molluscs were collected from five biotopes where rats had been found previously to harbor either adult worms or antibodies against A. cantonensis, and the identification was carried out on the basis of morphological features and a LAMP technique. The prevalence of A. cantonensis larvae in the mollusc samples, based on morphological identification, was 19.3%, whereas 59 out of the 98 individuals (60.2%) analyzed by LAMP were positive. Positive results were obtained for the three mollusc species analyzed and two of the positive samples, both obtained from P. lamarckii, were confirmed as positive by 18S rRNA and ITS1 PCR. Sequence analysis of 18S rRNA PCR products showed 100% similarity with previously published A. cantonensis sequences. These results may be relevant from a public health point of view, since all the biotopes from which the samples were obtained were in inhabited areas or areas with human activity, but it is also important from the perspective of a possible transmission to other accidental hosts, such as dogs and horses, animals that are present in some of the areas analyzed.