Pilar Foronda

Identification of genotypes of Giardia intestinalis of human isolates in Egypt

In Egypt, the genotyping study of Giardia intestinalis in human is limited. To determine the prevalence of G. intestinalis, faecal samples were collected from Egypt. Samples were concentrated using density gradient centrifugation. The samples were subjected to PCR and DNA sequence analysis for TPI gene. Prevalence of Giardia infection was 34.6% of 52 examined. DNA sequence showed that the Assemblage B was the most prevalent (80%) genotype, the 15% of the positive samples belonged to Assemblage E, and the 5% of them belonged to Assemblage A. Certainly, both genotypes A and B are highly common in human worldwide. However, up to now, Assemblage E had not been known to be infectious for humans. Therefore, this is the first time that Assemblage E is reported in human. However, further analyses of a second locus are required to confirm this result. The extent to which Giardia-infected cattle in Egypt might pose a risk of human infection is unknown.

Isolation and identification of pathogenic Acanthamoeba strains in Tenerife, Canary Islands, Spain from water sources

A comprehensive survey to document the presence of free-living amoebae of the genus Acanthamoeba was conducted in tap water and sea water sources related to human environments in Tenerife, Canary Islands, Spain. Acanthamoeba identification was based on the morphology of cyst and trophozoite forms and PCR amplification with a genus-specific primer pair. The pathogenic potential of Acanthamoeba isolates was characterized by temperature and osmotolerance assays and PCR reactions with two primer pairs related to Acanthamoeba pathogenesis. The results demonstrate the presence of potentially pathogenic strains in both sources. Thus, some of the amoebae in these aquatic habitats can act as opportunistic pathogens, could play a role in the diseases of aquatic organisms, and may present a risk to human health.

Occurrence and genotype characterization of Giardia duodenalis in goat kids from the Canary Islands, Spain

Giardia duodenalis (syn. Giardia lamblia, Giardia intestinalis) is a wide-spread intestinal protozoa of both humans and animals. Although giardiosis in goat is commonly asymptomatic, young kids may bear an enteric disease associated with persistent diarrhoea and delayed weight gain. In the present study we have analysed the occurrence of Giardia in 315 young goat kids (2-6 months old) from Gran Canaria Island (Spain) through visualization of faecal cysts. The identification of genotypes of G. duodenalis among the farms was attained by nested PCR of the triophosphate isomerase (TPI) and single PCR of beta-giardin genes and subsequent sequencing. Positive samples were found in 42.2% of the animals and 95.5% of the farms. Goat faecal specimens were positive for only livestock-associated G. duodenalis assemblage E genotype for both TPI and beta-giardin genes. The genetic analysis of these two loci revealed the presence of different haplotypes among the farms included in the survey and high homology with homologous genes from cattle and sheep. Altogether, the data presented here provide additional information to the prevalence and genetic characterization of Giardia isolates. The absence of assemblages A and B in this study suggests that zoonotic transmission of Giardia from goats could be of low epidemiological significance, although these findings should be validated in studies including other geographical areas, age groups and larger number of samples.

Systematic relationships of hymenolepidid cestodes of rodents and shrews inferred from sequences of 28S ribosomal RNA

Haukisalmi, V., Hardman, L. M., Foronda, P., Feliu, C., Laakkonen, J., Niemimaa, J., Lehtonen, J. T. & Henttonen, H. (2010). Systematic relationships of hymenolepidid cestodes of rodents and shrews inferred from sequences of 28S ribosomal RNA. —Zoologica Scripta, 39, 631–641.

Finding of Parastrongylus cantonensis (Chen, 1935) in Rattus rattus in Tenerife, Canary Islands (Spain)

Parastrongylus cantonensis is a parasite of murid rodents that can infect humans and cause health problems as eosinophilic meningitis. Although it is endemic in south Asia, the Pacific islands, Australia, USA, and a few Caribbean islands, it has been extended to new geographical regions. In the Canary Islands (Spain) a survey of helminths of Rattus rattus, Rattus norvegicus and Mus musculus domesticus was carried out. Furthermore, five species of molluscs were examined for nematode larvae to determine whether they are potential intermediate hosts of P. cantonensis. Nematodes were found in the lungs of 15% of 67 R. rattus examined in Tenerife, one of the four studied islands, with a prevalence of 20% in the highest focus of infection. Based on morphological and molecular analysis, with the complete internal transcribed spacer 2 (ITS-2) and a fragment of the small subunit ribosomal RNA (SSU rRNA) nucleotide sequences, nematodes were identified as P. cantonensis. Larval nematodes found from snails and slugs were identified as third-stage (L(3)) Metastrongyloidea, but the molecular study showed that they did not belong to P. cantonensis. This is the first finding of angiostrongyliasis in rats in the Canary Islands (Spain). New molecular data for this species and Parastrongylus dujardini are reported. The presence of P. cantonensis in Tenerife could be of importance from the public health point of view. Further studies are required in order to look for other potential foci of infections in the Canary Islands.

POPULATION DISTRIBUTION AND ZOONOTIC POTENTIAL OF GASTROINTESTINAL HELMINTHS OF WILD RATS RATTUS RATTUS AND R. NORVEGICUS FROM JAMAICA

The population distribution and zoonotic potential of gastrointestinal helminths in a naturally infected population of wild rats (Rattus rattus and Rattus norvegicus) in Jamaica are described. One hundred and thirty (29.7%) of 437 rats captured in the study were infected: 104 (35%) of 297 R. rattus compared with 26 (18.6%) of 140 R. norvegicus. Nine species of gastrointestinal helminths were recovered: Raillietina sp. (0.2%), Trichuris sp. (0.2%), Rictularia sp. (0.7%), Syphacia obvelata (1.1%), Strongyloides ratti (1.4%), Hymenolepis diminuta (3.8%), Protospirura muricola (4.3%), Moniliformis moniliformis (11.2%), and Nippostrongylus brasiliensis (14.2%). In a logistic model, the single risk factor identified for both M. moniliformis and P. muricola was R. rattus, compared with R. norvegicus (OR = 8.369 and 9.714, respectively). In comparison, the risk factor predicted for infection with N. brasiliensis was the northeastern section of Jamaica (OR = 11.000) compared with western Jamaica. Rictularia sp. represents a new geographic distribution record for the Caribbean region. Hymenolepis diminuta, M. moniliformis, Raillietina sp., and Rictularia sp. are potentially zoonotic, but only human infection with H. diminuta has been previously reported in the Caribbean.

HELMINTHS OF THE WILD RABBIT (ORYCTOLAGUS CUNICULUS) IN MACARONESIA

Two hundred and four rabbits from 8 Macaronesian islands (Pico, San Jorge, San Miguel, Terceira, and Flores from Azores Archipelago; Tenerife and Alegranza from Canary Islands; and Madeira from Madeira Archipelago) were examined for helminth parasites between 1995 and 2000. Three species of cestodes, Taenia pisiformis (larvae), Andrya cuniculi, and Mosgovoyia ctenoides, and 5 species of nematodes, Trichuris leporis, Graphidium strigosum, Trichostrongylus retortaeformis, Passalurus ambiguus, and Dermatoxys hispaniensis, were identified. Only 3 species (M. ctenoides, T. retortaeformis, and P. ambiguus) were regularly distributed over the 3 archipelagos. Taenia pisiformis was not collected in Madeira, nor was A. cuniculi in the Azores and G. strigosum in the Canary Islands. Trichuris leporis and D. hispaniensis were only found in Madeira. Significant differences in the general prevalence of the nematodes G. strigosum and T. retortaeformis were detected between Azores and Madeira. The prevalence of T. retortaeformis differs significantly between the Azores and the Canaries and that of P. ambiguus was higher in Madeira than in Azores and Canaries. The helminth richness found in the wild rabbit in these Macaronesian archipelagos was very low compared with the Palearctic helminth fauna of this host. The wild rabbit was introduced from the Iberian Peninsula into different Macaronesian islands. Helminths introduced with Oryctolagus cuniculus into these islands also are commonly found in Iberian wild rabbits, which are excellent colonizers, as demonstrated in this study.

Distribution and genetic variation of hymenolepidid cestodes in murid rodents on the Canary Islands (Spain)

BackgroundIn the Canary Islands there are no previous data about tapeworms (Cestoda) of rodents. In order to identify the hymenolepidid species present in these hosts, a survey of 1,017 murine (349 Rattus rattus, 13 Rattus norvegicus and 655 Mus musculus domesticus) was carried out in the whole Archipelago. Molecular studies based on nuclear ITS1 and mitochondrial COI loci were performed to confirm the identifications and to analyse the levels of genetic variation and differentiation.ResultsThree species of hymenolepidids were identified: Hymenolepis diminuta, Rodentolepis microstoma and Rodentolepis fraterna. Hymenolepis diminuta (in rats) and R. microstoma (in mice) showed a widespread distribution in the Archipelago, and R. fraterna was the least spread species, appearing only on five of the islands. The hymenolepidids found on Fuerteventura, Lanzarote and La Graciosa were restricted to one area. The COI network of H. diminuta showed that the haplotypes from Lanzarote and Fuerteventura are the most distant with respect to the other islands, but clearly related among them.ConclusionsFounder effects and biotic and abiotic factors could have played important role in the presence/absence of the hymenolepidid species in determined locations. The haplotypes from the eastern islands (Fuerteventura and Lanzarote) seem to have shared an ancestral haplotype very distant from the most frequent one that was found in the rest of the islands. Two colonization events or a single event with subsequent isolation and reduced gene flow between western-central and eastern islands, have taken place in the Archipelago. The three tapeworms detected are zoonotic species, and their presence among rodents from this Archipelago suggests a potential health risk to human via environmental contamination in high risk areas. However, the relatively low prevalence of infestations detected and the focal distribution of some of these species on certain islands reduce the general transmission risk to human.

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.

Proposal for a unified classification system and nomenclature of lagoviruses

Lagoviruses belong to the Caliciviridae family. They were first recognized as highly pathogenic viruses of the European rabbit (Oryctolagus cuniculus) and European brown hare (Lepus europaeus) that emerged in the 1970-1980s, namely, rabbit haemorrhagic disease virus (RHDV) and European brown hare syndrome virus (EBHSV), according to the host species from which they had been first detected. However, the diversity of lagoviruses has recently expanded to include new related viruses with varying pathogenicity, geographic distribution and host ranges. Together with the frequent recombination observed amongst circulating viruses, there is a clear need to establish precise guidelines for classifying and naming lagovirus strains. Therefore, here we propose a new nomenclature based on phylogenetic relationships. In this new nomenclature, a single species of lagovirus would be recognized and called Lagovirus europaeus. The species would be divided into two genogroups that correspond to RHDV- and EBHSV-related viruses, respectively. Genogroups could be subdivided into genotypes, which could themselves be subdivided into phylogenetically well-supported variants. Based on available sequences, pairwise distance cutoffs have been defined, but with the accumulation of new sequences these cutoffs may need to be revised. We propose that an international working group could coordinate the nomenclature of lagoviruses and any proposals for revision.