BackgroundDue to climate change, the geographical distribution of sand flies during the last decades has shifted northward from latitudes below 45°N in southern Europe to latitudes just above 50○N. Recent studies show that some phlebotomine sand flies were recorded in several parts of Germany and Belgium. In central Europe, some autochthone leishmaniasis cases are being recorded in regions traditionally regarded as leishmaniasis-free. An important challenge is to predict the geographical distribution of leishmaniasis vectors under new climatic conditions. In this study, we attempted to predict the current distribution of six leishmaniasis vectors in the Mediterranean basin and forecast species’ geographical shift under future climate scenarios using an ensemble ecological niche modeling approach. Species records were obtained from scientific surveys published in the research literature between 2006 and 2016. A series of climate metrics describing temperature and precipitation in the study area under two climatic scenarios were obtained from WorldClim database. A consensus model was derived from six varieties of modeling approaches (regression, machine learning and classification techniques) in order to ensure valid prediction of distribution of vectors under different climate scenarios.ResultsModel performance was generally high for the included species with a specificity (true negative rate) ranging from 81.03 to 96.52% (mean = 86.94%) and a sensitivity (true positive rate) ranging from 87.93 to 100% (mean = 96.98%). Our work evidenced the hypothesis of the widespread of Leishmania vectors under climate change scenarios. All of the studied species are prospected to gain new areas that are actually not suitable for vectors’ survival. Phlebotomine sand flies are prospected to invade extra-Mediterranean regions, especially western and central Europe.ConclusionsOur study confirmed the importance of environmental and climate factors on the distribution of leishmaniasis vectors and demonstrated the performance of ecological niche modeling in the prediction of the geographical spread of vector-borne diseases. Ecological niche modeling should be considered in the future as a valuable tool in addition to experimental laboratory studies for a better understanding of the biology of vector species.

Cutaneous leishmaniasis is a very complex disease involving multiple factors that limit its emergence and spatial distribution. Prediction of cutaneous leishmaniasis epidemics in Tunisia remains difficult because most of the epidemiological tools used so far are descriptive in nature and mainly focus on a time dimension. The purpose of this work is to predict the potential geographic distribution of Phlebotomus papatasi and zoonotic cutaneous leishmaniasis caused by Leishmania major in Tunisia using Grinnellian ecological niche modeling. We attempted to assess the importance of environmental factors influencing the potential distribution of P. papatasi and cutaneous leishmaniasis caused by L. major. Vectors were trapped in central Tunisia during the transmission season using CDC light traps (John W. Hock Co., Gainesville, FL). A global positioning system was used to record the geographical coordinates of vector occurrence points and households tested positive for cutaneous leishmaniasis caused by L. major. Nine environmental layers were used as predictor variables to model the P. papatasi geographical distribution and five variables were used to model the L. major potential distribution. Ecological niche modeling was used to relate known species' occurrence points to values of environmental factors for these same points to predict the presence of the species in unsampled regions based on the value of the predictor variables. Rainfall and temperature contributed the most as predictors for sand flies and human case distributions. Ecological niche modeling anticipated the current distribution of P. papatasi with the highest suitability for species occurrence in the central and southeastern part of Tunisian. Furthermore, our study demonstrated that governorates of Gafsa, Sidi Bouzid, and Kairouan are at highest epidemic risk.

Coxsackievirus B3 (CVB3) causes viral myocarditis and can ultimately result in dilated cardiomyopathy. There is no vaccine available for clinical use. Translation initiation of CVB3 RNA is directed by an internal ribosome entry site within the 5'-untranslated region. We have previously described that Sabin3-like mutation (U(473) to C) introduced in CVB3 genome led to a defective mutant with a serious reduction in translation efficiency. In the present study, we analyzed, in vitro, the effect of the Sabin3-like mutation on the binding affinity of RNA domain V to some standard translation initiation factors: eIF4G, eIF3b, and eIF4B by filter-binding assays and UV-crosslink assays. We have demonstrated that this single-nucleotide exchange impairs the binding affinity of these cellular factors within the mutant RNA. These data indicate how this decisive Sabin3-like mutation mediates viral translation attenuation. Taken together, these findings strongly suggest that the mutant strain could be considered a candidate for an attenuated CVB3 vaccine.

Influenza viruses are negative stranded segmented RNA viruses belonging to Orthomyxoviridae family. They are classified into three types A, B, and C. Type A influenza viruses are classified into subtypes according to the antigenic characters of the surface glycoproteins: hemagglutinin (H) and neuraminidase (N). The aim of the present study is to develop a fast and reliable multiplex RT-PCR technique for detecting simultaneously the subtypes A/H1N1 and A/H3N2 of influenza virus. Our study included 398 patients (mean age 30.33 ± 19.92 years) with flu or flu-like syndromes, consulting physicians affiliated with collaborating teams. A multiplex RT-PCR detecting A/H1N1 and A/H3N2 influenza viruses and an examination by indirect immunofluorescence (IFI) were performed. In the optimized conditions, we diagnosed by IFI a viral infection in 90 patients (22.6 %): 85 cases of influenza type A, four cases of influenza type B, and only one case of coinfection with types A and B. An evaluation of the technique was performed on 19 clinical specimens positive in IFI, and we detected eight cases of A/H3N2, five cases of A/H1N1, one case of influenza virus type A which is not an H1N1 nor H3N2, and five negative cases. Multiplex RT-PCR is a sensitive technique allowing an effective and fast diagnosis of respiratory infections caused by influenza viruses in which the optimization often collides with problems of sensibility.

In Tunisia, cases of zoonotic cutaneous leishmaniasis caused by Leishmania major are increasing and spreading from the south-west to new areas in the center. To improve the current knowledge on L. major evolution and population dynamics, we performed multi-locus microsatellite typing of human isolates from Tunisian governorates where the disease is endemic (Gafsa, Kairouan and Sidi Bouzid governorates) and collected during two periods: 1991–1992 and 2008–2012. Analysis (F-statistics and Bayesian model-based approach) of the genotyping results of isolates collected in Sidi Bouzid in 1991–1992 and 2008–2012 shows that, over two decades, in the same area, Leishmania parasites evolved by generating genetically differentiated populations. The genetic patterns of 2008–2012 isolates from the three governorates indicate that L. major populations did not spread gradually from the south to the center of Tunisia, according to a geographical gradient, suggesting that human activities might be the source of the disease expansion. The genotype analysis also suggests previous (Bayesian model-based approach) and current (F-statistics) flows of genotypes between governorates and districts. Human activities as well as reservoir dynamics and the effects of environmental changes could explain how the disease progresses. This study provides new insights into the evolution and spread of L. major in Tunisia that might improve our understanding of the parasite flow between geographically and temporally distinct populations.

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Background The ubiquitous environmental fungus Aspergillus flavus is also a life‐threatening avian pathogen. Objectives This study aimed to assess the genetic diversity and population structure of A. flavus isolated from turkey lung biopsy or environmental samples collected in a poultry farm. Methods A. flavus isolates were identified using both morphological and ITS sequence features. Multilocus microsatellite genotyping was performed by using a panel of six microsatellite markers. Population genetic indices were computed using FSTAT and STRUCTURE. A minimum‐spanning tree (MST) and UPGMA dendrogram were drawn using BioNumerics and NTSYS‐PC, respectively. Results The 63 environmental (air, surfaces, eggshells and food) A. flavus isolates clustered in 36 genotypes (genotypic diversity = 0.57), and the 19 turkey lung biopsies isolates clustered in 17 genotypes (genotypic diversity = 0.89). The genetic structure of environmental and avian A. flavus populations were clearly differentiated, according to both F ‐statistics and Bayesian model‐based analysis’ results. The Bayesian approach indicated gene flow between both A. flavus populations. The MST illustrated the genetic structure of this A. flavus population split in nine clusters, including six singletons. Conclusions Our results highlight the distinct genetic structure of environmental and avian A. flavus populations, indicative of a genome‐based adaptation of isolates involved in avian aspergillosis.

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