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Comparison of gene expression patterns among Leishmania braziliensis clinical isolates showing a different in vitro susceptibility to pentavalent antimony
Introduction. Evaluation of Leishmania drug susceptibility depends on in vitro SbV susceptibility assays, which are labour-intensive and may give a biased view of the true parasite resistance. Molecular markers are urgently needed to improve and simplify the monitoring of SbV-resistance. We analysed here the gene expression profile of 21 L. braziliensis clinical isolates in vitro defined as SbV-resistant and -sensitive, in order to identify potential resistance markers. Methods. The differential expression of 13 genes involved in SbV metabolism, oxidative stress or housekeeping functions was analysed during in vitro promastigote growth. Results. Expression profiles were up-regulated for 5 genes only, each time affecting a different set of isolates (mosaic picture of gene expression). Two genes, ODC (ornithine decarboxylase) and TRYR (trypanothione reductase), showed a significantly higher expression rate in the group of SbV-resistant compared to the group of SbV-sensitive parasites (P<0.01). However, analysis of individual isolates showed both markers to explain only partially the drug resistance. Discussion. Our results might be explained by (i) the occurrence of a pleiotropic molecular mechanism leading to the in vitro SbV resistance and/or (ii) the existence of different epi-phenotypes not revealed by the in vitro SbV susceptibility assays, but interfering with the gene expression patterns
Universal PCR assays for the differential detection of all Old World Leishmania species
For the epidemiological monitoring and clinical case management of leishmaniasis, determination of the causative Leishmania species gains importance. Current assays for the Old World often suffer from drawbacks in terms of validation on a geographically representative sample set and the ability to recognize all species complexes. We want to contribute to standardized species typing for Old World leishmaniasis. We determined the ribosomal DNA internal transcribed spacer 1 sequence of 24 strains or isolates, and validated four species-specific polymerase chain reactions (PCRs) amplifying this target. They discriminate L. aethiopica, L. tropica, L. major, and the L. donovani complex, use the same cycling conditions, and include an internal amplification control. Our PCRs amplify 0.1 pg of Leishmania DNA, while being 100% specific for species identification on an extensive panel of geographically representative strains and isolates. Similar results were obtained in an endemic reference laboratory in Kenya. Species could also be identified in clinical specimens. The presented PCRs require only agarose gel detection, and have several other advantages over many existing assays. We outline potential problems, suggest concrete solutions for transferring the technique to other settings, and deliver the proof-of-principle for analyzing clinical samples
Foot-and-Mouth Disease in Tanzania from 2001 to 2006
Foot-and-mouth disease (FMD) is endemic in Tanzania, with outbreaks occurring almost each year in different parts of the country. There is now a strong political desire to control animal diseases as part of national poverty alleviation strategies. However, FMD control requires improving the current knowledge on the disease dynamics and factors related to FMD occurrence so control measures can be implemented more efficiently. The objectives of this study were to describe the FMD dynamics in Tanzania from 2001 to 2006 and investigate the spatiotemporal patterns of transmission. Extraction maps, the space-time K-function and space-time permutation models based on scan statistics were calculated for each year to evaluate the spatial distribution, the spatiotemporal interaction and the spatiotemporal clustering of FMD-affected villages. From 2001 to 2006, 878 FMD outbreaks were reported in 605 different villages of 5815 populated places included in the database. The spatial distribution of FMD outbreaks was concentrated along the Tanzania-Kenya, Tanzania-Zambia borders, and the Kagera basin bordering Uganda, Rwanda and Tanzania. The spatiotemporal interaction among FMD-affected villages was statistically significant (P </= 0.01) and 12 local spatiotemporal clusters were detected; however, the extent and intensity varied across the study period. Dividing the country in zones according to their epidemiological status will allow improving the control of FMD and delimiting potential FMD-free areas
Drug susceptibility of Leishmania infantum (syn. Leishmania chagasi) isolates from Brazilian HIV-positive and HIV-negative patients
An outbreak of haemolytic anaemia associated with infection of Theileria orientalis in naive cattle
CASE HISTORY: An outbreak of haemolytic anaemia occurred when 87 cattle were introduced from a presumed non-infected herd from south Otago to a herd in Northland (n=580 cows), New Zealand, where theileriosis is endemic. CLINICAL FINDINGS: Clinical signs associated with Theileria spp. infection included lethargy, anorexia, inappetance, pale mucous membranes, and varying severity of anaemia. In the naive imported cattle, 11/29 (38%) of those tested showed haematological signs of anaemia (haematocrit (HCT) <0.25 L/L). A negative association was present between the HCT and the number of Theileria spp. organisms counted using light microscopy (correlation coefficient=-0.4; p<0.05). Haemoparasites consistent with Theileria spp. were observed on examination of a blood smear. Theileria orientalis group (Theileria buffeli/orientalis) species was confirmed using PCR and DNA sequencing, and other causes for anaemia were excluded in the most clinically severely affected cow. The 18S sequence data and phylogenetic analysis of the CoxIII sequences showed samples had the greatest similarity to T. orientalis Chitose from Japan. DIAGNOSIS: Haemolytic anaemia associated with infection of T. orientalis. CLINICAL RELEVANCE: Previous reports have suggested that T. orientalis group species may be non-pathogenic in healthy cattle, and an incidental finding in blood samples. However, this investigation provided evidence that in New Zealand, this pathogen is capable of causing clinical disease in cattle not necessarily debilitated by another disease. The potential for disease should be considered when naive cattle are brought in from non-endemic to endemic regions, for instance cattle from the South Island moved to regions where the vector for T. orientalis group species, Haemaphysalis longicornis, is active, and T. orientalis is present
Relative seroprevalence of cysticercus antigens and antibodies and antibodies to Taenia ova in a population sample in south India suggests immunity against neurocysticercosis
We evaluated the exposure of a community in Vellore district of south India to Taenia solium infection and its relationship to the prevalence of neurocysticercosis (NCC) causing active epilepsy. Seroprevalence of Taenia cysticercus antigens and antibodies were determined in 1064 randomly chosen asymptomatic individuals, antibodies to T. solium ova in 197 selected sera, and prevalence of taeniasis by a coproantigen test in 729 stool samples. The prevalence of NCC causing active epilepsy in Vellore district was determined in a population of 50 617. Coproantigens were detected in 0.8% (6 samples), Taenia cysticercus antigens in 4.5% (48 sera) and cysticercus IgG antibodies in 15.9% (169 sera) of the population. Cysticercus antibodies were directed against relatively low molecular weight cyst glycoprotein antigens in 14.9% (158 sera) of the population. IgG antibodies to Taenia ova were found in 81 (41.1%) of the selected samples. Prevalence of NCC causing active epilepsy was 1.3 per 1000 population. These results show high exposure of the population to the parasite and a relatively high prevalence of active infections (4.5% antigen positives) but a low prevalence of NCC causing active epilepsy (0.13%). These findings may indicate that the population is protected against developing neurocysticercosis. IgG antibodies directed against Taenia ova and low molecular weight cyst antigens may contribute to protection
Analysis of the editorial process of the multidisciplinary rural development journal Tropicultura
True versus apparent malaria infection prevalence: the contribution of a Bayesian approach
AIMS: To present a new approach for estimating the "true prevalence" of malaria and apply it to datasets from Peru, Vietnam, and Cambodia. METHODS: Bayesian models were developed for estimating both the malaria prevalence using different diagnostic tests (microscopy, PCR & ELISA), without the need of a gold standard, and the tests' characteristics. Several sources of information, i.e. data, expert opinions and other sources of knowledge can be integrated into the model. This approach resulting in an optimal and harmonized estimate of malaria infection prevalence, with no conflict between the different sources of information, was tested on data from Peru, Vietnam and Cambodia. RESULTS: Malaria sero-prevalence was relatively low in all sites, with ELISA showing the highest estimates. The sensitivity of microscopy and ELISA were statistically lower in Vietnam than in the other sites. Similarly, the specificities of microscopy, ELISA and PCR were significantly lower in Vietnam than in the other sites. In Vietnam and Peru, microscopy was closer to the "true" estimate than the other 2 tests while as expected ELISA, with its lower specificity, usually overestimated the prevalence. CONCLUSIONS: Bayesian methods are useful for analyzing prevalence results when no gold standard diagnostic test is available. Though some results are expected, e.g. PCR more sensitive than microscopy, a standardized and context-independent quantification of the diagnostic tests' characteristics (sensitivity and specificity) and the underlying malaria prevalence may be useful for comparing different sites. Indeed, the use of a single diagnostic technique could strongly bias the prevalence estimation. This limitation can be circumvented by using a Bayesian framework taking into account the imperfect characteristics of the currently available diagnostic tests. As discussed in the paper, this approach may further support global malaria burden estimation initiatives