María Dolores Bargues

Climate change effects on trematodiases, with emphasis on zoonotic fascioliasis and schistosomiasis.

The capacity of climatic conditions to modulate the extent and intensity of parasitism is well known since long ago. Concerning helminths, among the numerous environmental modifications giving rise to changes in infections, climate variables appear as those showing a greater influence, so that climate change may be expected to have an important impact on the diseases they cause. However, the confirmation of the impact of climate change on helminthiases has been reached very recently. Only shortly before, helminthiases were still noted as infectious diseases scarcely affected by climate change, when compared to diseases caused by microorganisms in general (viruses, bacteriae, protozoans). The aim of the present paper is to review the impact of climate change on helminthiases transmitted by snails, invertebrates which are pronouncedly affected by meteorological factors, by focusing on trematodiases. First, the knowledge on the effects of climate change on trematodiases in general is reviewed, including aspects such as influence of temperature on cercarial output, cercarial production variability in trematode species, influences of magnitude of cercarial production and snail host size, cercarial quality, duration of cercarial production increase and host mortality, influence of latitude, and global-warming-induced impact of trematodes. Secondly, important zoonotic diseases such as fascioliasis, schistosomiasis and cercarial dermatitis are analysed from the point of view of their relationships with meteorological factors. Emphasis is given to data which indicate that climate change influences the characteristics of these trematodiases in concrete areas where these diseases are emerging in recent years. The present review shows that trematodes, similarly as other helminths presenting larval stages living freely in the environment and/or larval stages parasitic in invertebrates easily affected by climate change as arthropods and molluscs as intermediate hosts, may be largely more susceptible to climate change impact than those helminths in whose life cycle such phases are absent or reduced to a minimum. Although helminths also appear to be affected by climate change, their main difference with microparasites lies on the usually longer life cycles of helminths, with longer generation times, slower population growth rates and longer time period needed for the response in the definitive host to become evident. Consequently, after a pronounced climate change in a local area, modifications in helminth populations need more time to be obvious or detectable than modifications in microparasite populations. Similarly, the relation of changes in a helminthiasis with climatic factor changes, as extreme events elapsed relatively long time ago, may be overlooked if not concretely searched for. All indicates that this phenomenon has been the reason for previous analyses to conclude that helminthiases do not constitute priority targets in climate change impact studies.

Mitochondrial 16S rDNA sequences and phylogenetic relationships of species of Rhipicephalus and other tick genera among Metastriata (Acari: Ixodidae)

Abstract The mitochondrial 16S rRNA gene sequences of the following eight European Metastriata tick species were obtained by direct polymerase-chain-reaction cycle sequencing and silver-staining methods: Rhipicephalus bursa, R. pusillus, R. sanguineus, R. turanicus, Boophilus annulatus, Dermacentor marginatus, Haemaphysalis punctata, and Hyalomma lusitanicum. This mitochondrial gene seems to be a good marker for the establishment of genetic relationships among closely related tick species, but it does not seem to be useful for comparisons of distantly related taxa. The molecular data provide very strong support for the monophyly of the Rhipicephalinae, including Hyalomma spp. However, the genus Rhipicephalus may not be considered a monophyletic group; in all analyses carried out in this study, R. bursa clustered with Boophilus spp. The high percentage of similarity (98.7%) observed between R.␣sanguineus and R. turanicus sequences would suggest that these species recently diverged within the Rhipicephalus genus. Phylogenetic analyses showed a monophyletic relationship among Amblyomminae taxa. The relationships between Haemaphysalis species and the true placement of this genus within Metastriata could not be resolved.

Phylogeography and Genetic Variation of Triatoma dimidiata, the Main Chagas Disease Vector in Central America, and Its Position within the Genus Triatoma

Background Among Chagas disease triatomine vectors, the largest genus, Triatoma, includes species of high public health interest. Triatoma dimidiata, the main vector throughout Central America and up to Ecuador, presents extensive phenotypic, genotypic, and behavioral diversity in sylvatic, peridomestic and domestic habitats, and non-domiciliated populations acting as reinfestation sources. DNA sequence analyses, phylogenetic reconstruction methods, and genetic variation approaches are combined to investigate the haplotype profiling, genetic polymorphism, phylogeography, and evolutionary trends of T. dimidiata and its closest relatives within Triatoma. This is the largest interpopulational analysis performed on a triatomine species so far. Methodology and Findings Triatomines from Mexico, Guatemala, Honduras, Nicaragua, Panama, Cuba, Colombia, Ecuador, and Brazil were used. Triatoma dimidiata populations follow different evolutionary divergences in which geographical isolation appears to have had an important influence. A southern Mexican–northern Guatemalan ancestral form gave rise to two main clades. One clade remained confined to the Yucatan peninsula and northern parts of Chiapas State, Guatemala, and Honduras, with extant descendants deserving specific status. Within the second clade, extant subspecies diversity was shaped by adaptive radiation derived from Guatemalan ancestral populations. Central American populations correspond to subspecies T. d. dimidiata. A southern spread into Panama and Colombia gave the T. d. capitata forms, and a northwestern spread rising from Guatemala into Mexico gave the T. d. maculipennis forms. Triatoma hegneri appears as a subspecific insular form. Conclusions The comparison with very numerous Triatoma species allows us to reach highly supported conclusions not only about T. dimidiata, but also on different, important Triatoma species groupings and their evolution. The very large intraspecific genetic variability found in T. dimidiata sensu lato has never been detected in a triatomine species before. The distinction between the five different taxa furnishes a new frame for future analyses of the different vector transmission capacities and epidemiological characteristics of Chagas disease. Results indicate that T. dimidiata will offer problems for control, although dwelling insecticide spraying might be successful against introduced populations in Ecuador.

The Northern Bolivian Altiplano: a region highly endemic for human fascioliasis.

SummaryThe worldwide importance of human infection by Fasciola hepatica has been recognized in recent years. The endemic region between Lake Titicaca and the valley of La Paz, Bolivia, at 3800–4100 m altitude, presents the highest prevalences and intensities recorded. Large geographical studies involving Lymnaea truncatula snails (malacological, physico‐chemical, and botanic studies of 59, 28 and 30 water bodies, respectively, inhabited by lymnaeids; environmental mean temperature studies covering a 40‐year period), livestock (5491 cattle) and human coprological surveys (2723 subjects, 2521 of whom were school children) were conducted during 1991–97 to establish the boundaries and distributional characteristics of this endemic Northern Altiplano region. The endemic area covers part of the Los Andes, Ingavi, Omasuyos and Murillo provinces of the La Paz Department. The human endemic zone is stable, isolated and apparently fixed in its present outline, the boundaries being marked by geographical, climatic and soil‐water chemical characteristics. The parasite distribution is irregular in the endemic area, the transmission foci being patchily distributed and linked to the presence of appropriate water bodies. Prevalences in school children are related to snail population distribution and extent. Altiplanic lymnaeids mainly inhabit permanent water bodies, which enables parasite transmission during the whole year. A confluence of several factors mitigates the negative effects of the high altitude.

Genomic changes of Chagas disease vector, South America.

We analyzed the main karyologic changes that have occurred during the dispersion of Triatoma infestans, the main vector of Chagas disease. We identified two allopatric groups, named Andean and non-Andean. The Andean specimens present C-heterochromatic blocks in most of their 22 chromosomes, whereas non-Andean specimens have only 4–7 autosomes with C-banding. These heterochromatin differences are the likely cause of a striking DNA content variation (approximately 30%) between Andean and non-Andean insects. Our study, together with previous historical and genetic data, suggests that T. infestans was originally a sylvatic species, with large quantities of DNA and heterochromatin, inhabiting the Andean region of Bolivia. However, the spread of domestic T. infestans throughout the non-Andean regions only involved insects with an important reduction of heterochromatin and DNA amounts. We propose that heterochromatin and DNA variation mainly reflected adaptive genomic changes that contribute to the ability of T. infestans to survive, reproduce, and disperse in different environments.

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.

Human and animal fascioliasis in Mazandaran province, northern Iran

The fascioliasis situation in humans and livestock of Iranian Mazandaran is analysed for the first time. Coprological studies showed 7.3% and 25.4% global prevalences in sheep and cattle, respectively. Studies in slaughterhouses indicate that sheep and cattle may be the main reservoir species, buffaloes may play local roles in the transmission, and goats and horses probably only participate sporadically. Morphometric studies by computer image analysis showed that forms intermediate between Fasciola hepatica and F. gigantica appear in addition to both species. A total of 107 infected humans were diagnosed during the 1999–2002 period, which suggests that fascioliasis may be widespread. The absence of differences in human fascioliasis among gender and age groups differs from other human endemic areas. Both human and animal infections show marked differences between western and eastern Mazandaran. Traditions in herbal condiments for human consumption, methods of animal husbandry and annual rainfall may explain the higher prevalences in western Mazandaran.

Populations, hybrids and the systematic concepts of species and subspecies in Chagas disease triatomine vectors inferred from nuclear ribosomal and mitochondrial DNA.

In Chagas disease, triatomine vectors are the main target for control measures because of the absence of effective drugs. The broad usefulness of nuclear rDNA and mtDNA sequences explains why triatomine studies using these markers have increased so pronouncedly in recent years. This indicates the appropriateness of an updated review about these molecular markers, concentrating on aspects useful for research on Chagas disease vectors. A comparative analysis is presented on the efficiency, weight of their different characteristics, limitations and problems of each of the different DNA markers in the light of the results obtained in studies on populations, hybrids, subspecies and species of the subfamily Triatominae. The use of a standardized composite haplotype code nomenclature for both nuclear rDNA and mtDNA markers is strongly encouraged to avoid difficulties in comparative studies. Triatomine aspects related to concerted evolution, microsatellites, minisatellites and insertions/deletions in nuclear rDNA and silent/non-silent mutations, pseudogenes and weaknesses of partial sequences in mtDNA are analysed. Introgression and hybrids, nuclear and mitochondrial DNA strengths, and compared evolutionary rates of nuclear rDNA and mtDNA in triatomines are discussed. Many conclusions are obtained thanks to the availability, for the first time in triatomines, of a complete sequence of a protein-coding mtDNA gene as ND1 from very numerous triatomine species covering from different populations of a species up to members belonging to different tribes. The evolutionary rates of each nuclear rDNA marker and mtDNA marker are analysed by comparison at subspecies level (intrapopulational, interpopulational, between morphs, and between subspecies) and species level (close and distant species of the same genus, species of different genera, and species of different tribes). Weaknesses of mtDNA for systematic-taxonomic purposes detected recently and newly in insects and triatomines, respectively, are discussed in detail. Emphasis is given to taxonomic units and biological entities presenting well-known problematics, both from the systematic-taxonomic and/or epidemiological-control points of view, as well as to molecular situations which can give rise to erroneous conclusions. All these aspects constitute the background on which the key question about the systematic concepts of species and subspecies in triatomines is focused. The global purpose is to facilitate future work on triatomines by highlighting present gaps, how better choice the appropriate markers, and marker aspects which should be taken into account. Key characteristics as alpha, CI and transformation rate matrices ought to be obtained and noted to get appropriate results and allow correct interpretations. The main aim is to offer a baseline for future fundamental research on triatomines and applied research on transmission, epidemiology and control measures related to Chagas disease vectors.

18S rRNA gene sequences and phylogenetic relationships of European hard-tick species (Acari: Ixodidae)

Abstract The complete 18S rRNA gene sequences of the following six European hard-tick species were obtained by direct PCR cycle sequencing and silver-staining methods: Rhipicephalus pusillus, Boophilus annulatus, Dermacentor marginatus, Hyalomma lusitanicum, Haemaphysalis punctata, and Ixodes ricinus. Differences observed in the sequence alignment of these six species together with the 18S rRNA gene sequences of 13 other hard-tick species demonstrate that this gene is a good marker for supraspecific differentiation as well as genus grouping among hard ticks. Phylogenetic analyses strongly support that Hyalomma species share a common ancestor with Rhipicephalinae and, consequently, Hyalomminae should no longer be considered an independent subfamily. However, no definitive conclusion could be reached to support or oppose the separation of the subfamilies Haemaphysalinae and Amblyomminae.

Phylogeography and genetic divergence of some lymnaeid snails, intermediate hosts of human and animal fascioliasis with special reference to lymnaeids from the Bolivian Altiplano

A population genetic study using starch gel electrophoresis was performed on populations of several species of lymnaeid snails acting as intermediate hosts for Fasciola hepatica (Trematoda, Plathyhelminth). Lymnaea viatrix was collected in 16 sites from the Bolivian Northern Altiplano. L. cubensis were obtained in one site from Venezuela, one site from Guadeloupe, three sites from Cuba and one site from the Dominican Republic. L. truncatula were collected in one site from France, one from Portugal and one from Morocco. Multilocus enzyme electrophoresis (MEE) were determined for 282 snails at 18 loci. A complete monomorphism was encountered at each geographic site. However, among these 18 loci, 13 are polymorphic and low and high levels of genetic divergence were observed between samples. Two genotypic groups can be differentiated by their multilocus genotypes. The western genotypic group associates together samples from Venezuela, Guadeloupe, Cuba and Dominican Republic (L. cubensis) while samples from France, Portugal and Morocco (L. truncatula) belong to the eastern genotypic group. Surprisingly, the Northern Bolivian Altiplano populations (L. viatrix) do not present any genetic divergence with the Portuguese sample. Therefore, the Bolivian snails belong entirely to the eastern genetic group. Within each group slight genetic divergences were observed. These results strongly support the European origin of the lymnaeid snails from the Northern Bolivian Altiplano.