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Background: Q fever is an underreported zoonotic disease of cattle and men in most countries of the world. Very little information about the prevalence of Coxiella burnetii infection in animals and humans comes from South and Central America and systematic studies are lacking. Methods: A seroprevalence survey for Q fever amongst cattle, farm workers and students was conducted in Ecuador using a commercial ELISA kit. Results: Survey results showed an unexpectedly high prevalence of Coxiella burnetii antibodies in dairy cattle (43%) and in farm workers (34%). In addition, a clinical case in a human of acute Q fever in the convalescent stage was detected. Conclusion: We conclude that the disease is endemic in Ecuador but is overlooked by medical and laboratory personnel. Q fever should be considered a public health issue in Ecuador and further research into the clinical relevance of this infection is recommended.

Cryptic species can present a significant challenge to the application of systematic and biogeographic principles, especially if they are invasive or transmit parasites or pathogens. Detecting cryptic species requires a pluralistic approach in which molecular markers facilitate the detection of coherent taxonomic units that can then be analyzed using various traits (e.g., internal morphology) and crosses. In asexual or self-fertilizing species, the latter criteria are of limited use. We studied a group of cryptic freshwater snails (genusGalba)from the family Lymnaeidae that have invaded almost all continents, reproducing mainly by self-fertilization and transmitting liver flukes to humans and livestock. We aim to clarify the systematics, distribution and phylogenetic relationships of these species with an integrative approach that includes morphology (shell and reproductive anatomy), molecular markers, wide-scale sampling across America, and data retrieved from GenBank (to include Old World samples). Our phylogenetic analysis suggests that the genusGalbaoriginated ca. 22 Myr ago and today comprises six clusters of species. Four of these clusters (G.truncatula,G.cubensis/viator,G.humilisandG.schirazensis) are morphologically cryptic and constitute species or species complexes with wide variation in their genetic diversity, geographic distribution and invasiveness. The other two clusters constitute a single species or a species complex (Galba cousini/meridensis) that demonstrate more geographically restricted distributions and exhibit an alternative morphology more phylogenetically derived than the cryptic one. Further genetic studies are required to clarify the status of bothG. cousini/meridensisandG.cubensis/viator. We emphasize that noGalbaspecies should be identified without molecular markers and that additional sampling is required, especially in North America, Eurasia and Africa to clarify remaining questions in systematics and biogeography. We also discuss several hypotheses that can explain crypsis inGalba, such as convergence and morphological stasis, and hypothesize a role for stabilizing selection in amphibious (rather than strictly freshwater) habitats.

The aim of the present study was to assess the resistance status of bovine gastrointestinal nematodes (GINs) against ivermectin (IVM) and fenbendazole (FBZ) in Ecuador. The study involved five cattle farms located in different topographic zones of the country. Anthelmintic efficacy was assessed by calculating the percentage of fecal egg counts reduction (FECR) after treatment. Additionally, DNA from pooled larval cultures was screened to ascertain benzimidazole resistance alleles. For animals treated with IVM, FECR percentages ranged from 0 to 68%, indicating the presence of highly resistant worms. The opposite was found for animals treated with FBZ, where FECR percentages were above 90% on all the farms tested. Pooled coprocultures revealed that Cooperia spp. were the predominant species pre and post-treatment although minor proportions of Haemonchus spp. and Ostertagia spp. were also identified. No mutations conferring resistance to benzimidazoles were identified in the beta-tubulin isotype 1 gene of the isolated Cooperia spp. worms, which is in line with the results of the FECR performed with FBZ. Overall, the present study highlights widespread resistance of bovine GINs to IVM but no to FBZ in Ecuador.

Some Lymnaeid snails are intermediate hosts of the liver fluke Fasciola hepatica, the causal agent of fasciolosis, a zoonotic parasitic disease. Human and livestock fasciolosis has been reported in a highland community located in the Chimborazo Province of the Ecuadorian Andes. However, no previous study has been carried out to identify which snail species act as intermediate host/s of F. hepatica. This study first aimed to identify the intermediate snail species and secondly to determine the prevalence of natural infection with F. hepatica in 230 lymnaeid snails sampled from irrigation and drainage canals in this area. The first objective entailed observations of shell morphology and internal organs as well as sequencing of the cytochrome oxidase subunit 1 (COI) gene. For the second objective, we used classic parasitological methods (observation of rediae and cercarial emission) and PCR amplification specie-specific to F. hepatica. COI haplotype networks were built to elucidate phylogeographic relationships between the snail populations from this highland community with other American and worldwide populations. We identified two lymnaeid Galba cousini and Galba schirazensis and found high infection rates of F. hepatica in G. cousini, but these differed according to the method used, with PCR showing a higher rate (61 ± 20%) compared to rediae observation (29 ± 17%). F. hepatica in G. schirazensis was identified only by DNA amplification. G. cousini populations were genetically structured by geographic distance whereas G. schirazensis populations showed very low genetic diversity. The higher abundance and infection rate of G. cousini compared to G. schirazensis suggests that the former is likely the specie responsible for F. hepatica transmission in this region.

We studied the population genetic structure of the freshwater snail Galba schirazensis (Küster, 1862), a potential vector of infectious diseases such as fascioliasis. Galba schirazensis has now a worldwide distribution but a poorly known origin because this species has been distinguished only recently from the morphologically similar and cosmopolitan Galba truncatula (O.F. Müller, 1774). We developed specific microsatellite markers and sequenced a mitochondrial gene (cytochrome oxidase subunit I (CO1)) to study individuals of G. schirazensis from the Old World and the New World. We found very low genetic diversity within populations, no heterozygotes, and marked population structure — a pattern observed in other highly selfing lymnaeid species with recently enlarged distributions as a result of biological invasions. The total lack of observed heterozygosity in the few populations of G. schirazensis that displayed some allelic diversity suggests high selfing rates. We also found that the center of diversity, and by extension the origin area of this species, should be found in the New World, whereas Old World populations should rather result from a recent introduction of a genetically uniform population. The microsatellite markers developed here will help to clarify the history of expansion of G. schirazensis and might help to understand its role as a potential vector of infectious diseases. Fil: Lounnas, Manon. Université de Montpellier. Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle; Francia. Centre National de la Recherche Scientifique. Institut de Recherche pour le Développement; Francia Fil: Correa, Ana Cristina. Centre National de la Recherche Scientifique. Institut de Recherche pour le Développement; Francia. Université de Montpellier. Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle; Francia Fil: Alda, Maria del Pilar. Centre National de la Recherche Scientifique. Institut de Recherche pour le Développement; Francia. Université de Montpellier. Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentina Fil: David, Patrice. Université de Montpellier. Centre d’Ecologie Fonctionnelle et d’Evolution; Francia Fil: Dubois, Marie Pierre. Université de Montpellier. Centre d’Ecologie Fonctionnelle et d’Evolution; Francia Fil: Calvopiña, Manuel. Universidad Central del Ecuador, Carrera de Medicina; Ecuador Fil: Caron, Yannick. Université de Liège; Bélgica Fil: Celi Erazo, Maritza. Universidad Central del Ecuador; Ecuador Fil: Bui, Dung Thi. Université de Liège; Bélgica Fil: Jarne, Philippe. Université de Montpellier. Centre d’Ecologie Fonctionnelle et d’Evolution; Francia Fil: Loker, Eric. University Of New Mexico. Department Of Biology; Estados Unidos Fil: Noya, Oscar. Universidad Central de Venezuela; Venezuela Fil: Rodríguez Hidalgo, Richar. Universidad Central del Ecuador; Ecuador Fil: Toty, Céline. Université de Montpellier. Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle; Francia Fil: Uribe, Nelson. Universidad Industrial Santander; Colombia Fil: Pointier, Jean Pierre. Université de Perpignan; Francia. Centre National de la Recherche Scientifique; Francia Fil: Hurtrez Boussès, Sylvie. Université de Montpellier. Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle; Francia

Currently, there is a lack of surveys that report the occurrence of gastrointestinal parasites in the white-headed capuchin monkey (Cebus albifrons). We therefore assessed the presence and richness (= number of different parasite genera) of parasites in C. albifrons in wildlife refuges (n = 11) and in a free-ranging group near a human village (n = 15) in the Ecuadorian Amazon. In the 78 samples collected (median of 3 samples per animal), we identified a total of 6 genera of gastrointestinal parasites, representing protozoa, nematodes, acanthocephalans and cestodes. We observed a high prevalence (84%) across the 26 individuals, with the most prevalent parasite being Strongyloides sp. (76.9%), followed by Hymenolepis sp. (38.5%) and Prosthenorchis elegans (11.5%). We found Entamoeba histolytica/dispar/moskovskii/nuttalli and Capillaria sp. in only a minority of the animals (3.8%). In addition, we observed unidentified strongyles in approximately one-third of the animals (34.6%). We found a total of 6 parasite genera for the adult age group, which showed higher parasite richness than the subadult age group (5) and the juvenile age group (3). Faecal egg/cyst counts were not significantly different between captive and free-ranging individuals or between sexes or age groups. The free-ranging group had a higher prevalence than the captive group; however, this difference was not significant. The only genus common to captive and free-ranging individuals was Strongyloides sp. The high prevalence of gastrointestinal parasites and the presence of Strongyloides in both populations support results from previous studies in Cebus species. This high prevalence could be related to the high degree of humidity in the region. For the free-ranging group, additional studies are required to gain insights into the differences in parasite prevalence and intensity between age and sex groups. Additionally, our study demonstrated that a serial sampling of each individual increases the test sensitivity., Graphical abstract Image 1, Highlights • This study brings new information about primate parasites to Ecuador. • We analyse the prevalence, richness and FECC among the two habitats, sex and age. • Adults harbour more genera of parasites than subadults and juveniles. • Sampling effort was essential to find a good number of parasites genera. • E. histolytica/dispar and P. elegans were found only in captive individuals.

Brucellosis is an important but neglected zoonosis that causes serious economic losses both in livestock and human populations. The aim of the present study was to estimate the true prevalence of brucellosis together with diagnostic sensitivity and specificity of three serological tests in humans of the northwestern part of Ecuador using a Bayesian approach adjusted for the dependencies among the multiple tests to avoid any misinterpretation. In addition, the causal agent responsible for human brucellosis was also identified. Using a total of 3,733 samples collected from humans in this area between 2006 and 2008, the prevalence of human brucellosis and the diagnostic test characteristics of the Rose Bengal fast agglutination test (RBT), Wright’s slow agglutination test with ethylenediaminetetraacetic acid disodium salt dehydrate (EDTA) (SAT-EDTA), and indirect ELISA (iELISA) were estimated using a Bayesian approach. The estimated true prevalence of human brucellosis was 1% (credibility interval: 0.4–1.6). The sensitivities of iELISA and RBT were higher than and similar (95.1% and 95.0%, respectively) to those of SAT-EDTA (60.8%). Even though all tests indicated a high specificity (> 99.0%), the specificity of SAT-EDTA was highest (99.9%). The circulating strain in this study area was identified to be Brucella abortus biotype 4 based on culture and microbiological characterization. The RBT and the iELISA are recommended for estimating the true prevalence of human brucellosis and/or for surveillance programs following their high sensitivities and specificities. The proposed strategy supports evidence-based medicine for clinicians and policy-makers to ensure appropriate preventive and control program of brucellosis worldwide.

Background: Q fever is an underreported zoonotic disease of cattle and men in most countries of the world. Very little information about the prevalence of Coxiella burnetii infection in animals and humans comes from South and Central America and systematic studies are lacking. Methods: A seroprevalence survey for Q fever amongst cattle, farm workers and students was conducted in Ecuador using a commercial ELISA kit. Results: Survey results showed an unexpectedly high prevalence of Coxiella burnetii antibodies in dairy cattle (43%) and in farm workers (34%). In addition, a clinical case in a human of acute Q fever in the convalescent stage was detected. Conclusion: We conclude that the disease is endemic in Ecuador but is overlooked by medical and laboratory personnel. Q fever should be considered a public health issue in Ecuador and further research into the clinical relevance of this infection is recommended.

International audience; A molecular tool described here allows in one step for specific discrimination among three cryptic freshwater snail species (genus Galba) involved in fasciolosis transmission, a worldwide infectious disease of humans and livestock. The multiplex PCR approach taken targets for each species a distinctive, known microsatellite locus which is amplified using specific primers designed to generate an amplicon of a distinctive size that can be readily separated from the amplicons of the other two species on an agarose gel. In this way, the three Galba species (G. cubensis, G. schirazensis, and G. truncatula) can be differentiated from one another, including even if DNA from all three were present in the same reaction. The accuracy of this new molecular tool was tested and validated by comparing multiplex PCR results with species identification based on sequences at mitochondrial and nuclear markers. This new method is accurate, inexpensive, simple, rapid, and can be adapted to handle large sample sizes. It will be helpful for monitoring invasion of Galba species and for developing strategies to limit the snail species involved in the emergence or re-emergence of fasciolosis.

International audience; Fasciolosis is a widely distributed disease in livestock in South America but knowledge about the epidemiology and the intermediate hosts is relatively scarce in Ecuador. For three months, lymnaeid snails were sampled (n = 1482) in Pichincha Province at two sites located in a highly endemic area. Snails were identified (based on morphology and ITS-2 sequences) and the infection status was established through microscopic dissection and a multiplex polymerase chain reaction (PCR)-based technique. Techniques based on morphology were not useful to accurately name the collected snail species. Comparison with available DNA sequences showed that a single snail species was collected, Galba schirazensis. Live rediae were observed in 1.75% (26/1482) and Fasciola sp. DNA was detected in 6% (89/1482) of collected snails. The COX-1 region permitted identification of the parasite as Fasciola hepatica. The relative sensitivity and specificity of the microscope study, compared to PCR results, were 25.84% and 99.78%, respectively. The mean size of the snails recorded positive for F. hepatica through crushing and microscopy was significantly higher than the mean size of negative snails, but there was no such difference in PCR-positive snails. The role of G. schirazensis as an intermediate host of F. hepatica in Ecuador is discussed and the hypothesis of an adaptation of the parasite to this invasive snail is proposed. For the first time, an epidemiological survey based on molecular biology-based techniques assessed the possible role of lymnaeid snails in the epidemiology of fasciolosis in Ecuador; La fasciolose est une maladie largement distribuée chez le bétail en Amérique du Sud, mais les connaissances sur l'épidémiologie et les hôtes intermédiaires sont relativement rares en Équateur. Pendant trois mois, les Lymnaeidae ont été échantillonnés (n = 1482) dans la province de Pichincha dans deux sites situés dans une zone fortement endémique. Les mollusques ont été identifiés (en fonction de la morphologie et des séquences ITS-2) et l'état de l'infection a été établi grâce à une dissection microscopique et une technique de PCR multiplex. Les techniques basées sur la morphologie n'ont pas été utiles pour identifier avec précision l'espèce de mollusque récoltée. Une comparaison avec les séquences d'ADN disponibles a montré qu'une seule espèce de mollusque a été récoltée, Galba schirazensis. Parmi les mollusques collectés, des rédies vivantes ont été observées chez 1.75 % (26/1482) et de l'ADN de Fasciola sp. été détecté dans 6 % (89/1482). La région COX-1 a permis d'identifier le parasite comme Fasciola hepatica. La sensibilité relative et la spécificité des études microscopiques, en comparaison avec les résultats de la PCR, étaient respectivement de 25.84 % et 99.78 %. La taille moyenne des mollusques considérés positifs pour F. hepatica par écrasement et microscopie était significativement plus élevée que la taille moyenne des mollusques négatifs, mais une telle différence n'a pas été trouvée chez les mollusques PCR-positifs. Le rôle de G. schirazensis en tant qu'hôte intermédiaire de F. hepatica en Équateur est discuté et l'hypothèse d'une adaptation du parasite à ce mollusque invasif est proposée. Pour la première fois, une enquête épidémiologique, basée sur des techniques de biologie moléculaire, a évalué le rôle possible des mollusques Lymnaeidae dans l'épidémiologie de la fasciolose en Équateur.

Background Human cysticercosis is a zoonotic disease causing severe health disorders and even death. While prevalence data become available worldwide, incidence rate and cumulative incidence figures are lacking, which limits the understanding of the Taenia solium epidemiology. Methodology/Principal findings A seroepidemiological cohort study was conducted in a south-Ecuadorian community to estimate the incidence rate of infection with and the incidence rate of exposure to T. solium based on antigen and antibody detections, respectively. The incidence rate of infection was 333.6 per 100,000 person-years (95% CI: [8.4–1,858] per 100,000 person-years) contrasting with a higher incidence rate of exposure 13,370 per 100,000 person-years (95% CI: [8,730–19,591] per 100,000 person-years). The proportion of infected individuals remained low and stable during the whole study year while more than 25% of the population showed at least one antibody seroconversion/seroreversion during the same time period. Conclusions/Significance Understanding the transmission of T. solium is essential to develop ad hoc cost-effective prevention and control programs. The estimates generated here may now be incorporated in epidemiological models to simulate the temporal transmission of the parasite and the effects of control interventions on its life cycle. These estimates are also of high importance to assess the disease burden since incidence data are needed to make regional and global projections of morbidity and mortality related to cysticercosis., Author Summary Human cysticercosis is a neglected zoonotic parasitic disease causing severe health disorders such as epilepsy and even death. Cysticercosis is related to poverty, inadequate hygiene conditions and traditional pig farming. The present study describes the dynamic nature of human Taenia solium larval infections in an Ecuadorian endemic community. In this study we report for the first time incidence rate and cumulative incidence figures of human T. solium larval infections in Latin America. The simultaneous use of antibody and antigen serological detections allowed estimating both parasite exposure and infection rates, respectively. While about 13% of the inhabitants were exposed to T. solium eggs, less than 1% of the population became yearly infected with the parasite. This contrast between exposure and infection may be linked to an effective resistance to the parasite acquired through long-term exposure of the population and differs from the African situation, where much higher levels of infection have been observed. These estimates are of high importance to understand the epidemiology of T. solium in order to develop ad hoc cost-effective prevention and control programs. They are also essential to assess the burden of T. solium cysticercosis since longitudinal data are needed to make regional and global projections of morbidity and mortality related to cysticercosis.

(Neuro)cysticercosis is an important zoonotic disease caused by infection with Taenia solium metacestode larvae. Existing immunodiagnostic techniques detect antibodies and circulating antigens (Ag) in serum and cerebrospinal fluid (CSF). Blood/CSF collection is an invasive procedure associated with blood-borne infections and is often not well accepted by communities. Detection of circulating Ag in urine has been suggested as an alternative, however this has been evaluated in clinical settings only. The aim of the present study was to evaluate the performance of a urine Ag-ELISA under field conditions. Paired serum and urine samples were obtained from participants in endemic areas of Ecuador (n=748) and Zambia (n=690) and were subjected to a monoclonal antibody-based Ag-ELISA. Calculation of positive and negative agreement indices (AI) showed better agreement in the negative direction both for Ecuadorian and Zambian samples (AI of 93.1 and 86.8, respectively). Using a Bayesian approach to determine the test characteristics, similar sensitivities were obtained for serum and urine Ag detection, whereas a decreased specificity was determined for the urine Ag-ELISA with a lower specificity (78.6%) for Zambian samples than for Ecuadorian samples (88.4%). This study indicates a higher specificity for the serum test under field conditions and promotes further research to improve the urine test.

We studied the prevalence of bovine tuberculosis (BTB) in Mejia canton, the major dairy cattle production area in northern Ecuador. Twenty dairy herds comprising 2,022 cattle were selected. In 2007, each animal was tested using the comparative intradermal tuberculin test (CITT). In 2008, a follow-up test was performed in the same herds. The true annual incidence was 1.70%, and the true prevalence was 7.41% and 7.13% in 2007 and 2008, respectively. The prevalence was 0.27% and 0.57% in medium-sized herds in 2007 and 2008, respectively, compared with 8.63% and 8.43% in large herds (P < 0.01). The number of skin testpositive cases also increased significantly with age (P = 0.03), contacts with other species of animals (P < 0.01), and introduction of new cattle (P = 0.04). Herd prevalence was 55% in 2007 and 65% in 2008. This study shows the lack of knowledge in cattle farmers about this zoonosis and the necessity for a national BTB control program in Ecuador.

Taenia solium is endemic in the Andean region of Ecuador. The recent rediscovery of Taenia saginata in humans urges to reconsider some assumptions in relation to the epidemiology of the taeniosis/cysticercosis complex in this country. Therefore, data were compiled on the infection of both tapeworms in man and animals in Pichincha and Imbabura provinces in the Andean region, north of Quito. On post mortem inspection 3 out of 806 (0.37%) carcasses had T. saginata metacestodes, however, 35 sera out of 869 (4.03%) showed circulating antigen in a monoclonal antibody-based sandwich ELISA (Ag-ELISA). Porcine cysticercosis was detected in 15 out of 2896 (0.52%) carcasses and 93 out of 1032 serum samples (9.01%) were positive in Ag-ELISA. In humans, 4.99% (215 out of 4306) cases of antigen positives were found, whereas coprological examination of 1935 stools resulted in 30 positive cases (1.55%). The limited number of adult tapeworms (29) that were collected does not allow firm conclusions on the proportion of each species, but in total 21 specimen were identified as T. saginata and 8 as T. solium . These data have been discussed in view of the epidemiology of human cysticercosis.

This collection of articles provides an account of the papers delivered at the 19th International Conference of the World Association for the Advancement of Veterinary Parasitology (WAAVP) (held in New Orleans, LA, USA, from 10 to 14 August 2003) in a symposium session on assessing the burden of Taenia solium cysticercosis and echinococcosis organised and chaired by A. Lee Willingham III from the WHO/FAO Collaborating Center for Research and Training on Emerging and other Parasitic Zoonoses in Denmark and Peter M. Schantz from the Parasitic Diseases Division of the US Centers for Disease Control and Prevention, USA. The focus was on the persistence of the zoonotic parasitic diseases cysticercosis, caused by the pork tapeworm T. solium, and echinococcosis, caused by species of the tapeworm Echinococcus, and why these diseases are given very little attention on the national and international agendas in spite of the availability of tools to detect, treat, control and prevent them when it is quite clear in most instances that they are clearly associated with and help perpetuate poverty. A major reason for this is that in many endemic areas the presence and impact of these diseases are not known due to the lack of investigation and information thus policy makers are not aware of their burden and benefits of their control. Documentation is also needed to help increase awareness of the international community and hopefully result in financial and technical support being made available. Thus, burden assessments of cysticercosis and echinococcosis provide an essential evidence base for securing political will and financial and technical support as well as providing a basis for cost-benefit analysis of prevention and control efforts. In order to make an appropriate and full burden assessment one must consider the health, agricultural, social and other impacts of these parasitic zoonoses comprehensively. During the symposium presentations were given concerning current ongoing initiatives to assess the burden of cysticercosis and echinococcosis and examples of the impact of these diseases in both developing and developed countries were provided. In addition, cost factors related to vaccines for these cestode diseases were discussed and the possibilities for technical and financial support from multilateral agencies for assessments and interventions presented.