Abstract Background Diarrhea is a leading cause of mortality in children under five, with rotavirus and enteric adenoviruses F40/F41 being significant contributors. In regions like Africa and Southeast Asia, the incidence is exacerbated by poor hygiene and sanitation, as well as the challenges around availability and affordability of healthcare and disease diagnostics. The objective of this systematic review was to identify and evaluate molecular assays for the detection of rotavirus and enteric adenoviruses F40/F41, with particular emphasis on diagnostic performance and suitability for point-of-care use. Methods A systematic review of literature from 2001 to 2024 was conducted following PRISMA guidelines. Out of 659,613 records screened, a subset of primary studies reporting on molecular assays development or diagnostic performance met the inclusion criteria. Studies exclusively focused on epidemiology, vaccine impact, or narrative reviews were excluded from the analytical synthesis. Results The review identified 26 PCR-based assays and 3 isothermal amplification assays. Reported clinical sensitivities ranged from approximately 85% to 100%, with specificities generally above 95% across studies. Several assays also demonstrated low limits of detection and turnaround times compatible with decentralized use. In particular, three LAMP assays and three PCR-based methods fulfilled key performance and operational criteria for point-of-care diagnostics. The review concluded that molecular diagnostics, especially LAMP assays, are effective for rapid detection of rotavirus and enteric adenoviruses F40/F41. Conclusion While PCR assays are considered the gold standard, LAMP assays are more accessible and faster, making them better suited for use in resource-limited settings. These assays show promising performance and may support point-of-care diagnostics.

To overcome the limitation of lateral flow immunoassays (LFIAs) in multiplexing while preserving high sensitivity and specificity, we present COLPLEX, a high-affinity recognition system based on the ultra-specific interaction between colicin antimicrobial proteins and their cognate immunity proteins. These protein pairs, naturally evolved to exhibit femtomolar binding affinities, are bio-orthogonal and readily accessible through recombinant expression. We integrated the COLPLEX system into a biplex LFIA platform for the simultaneous detection of glycoproteins from Zaire and Sudan Ebola virus variants. Using two distinct, non-cross-reactive colicin-immunity pairs, we immobilized colicins on the nitrocellulose membrane and fused immunity proteins to the capture antibodies. This configuration significantly improved the signal-to-noise ratio, reduced capture antibody consumption, and minimized non-specific binding. Clinical evaluation using sera from Ebola-infected patients (n = 30) and negative controls (n = 40), including healthy donors and patients with other viral haemorrhagic fevers, demonstrated high diagnostic accuracy, with 96.7 % sensitivity and 100 % specificity, confirming the robustness of COLPLEX for real-world applications. These results highlight COLPLEX as a powerful alternative to streptavidin-biotin system and its potential to revolutionize next-generation LFIA platforms.

Background Sub-Saharan Africa faces persistent challenges in the timely detection of infectious disease outbreaks due to inadequate early warning and response systems. To address this gap, Senegal's Ministry of Health partnered with the Institut Pasteur de Dakar to establish the Senegalese Syndromic Sentinel Surveillance Network (4S Network) in 2012 a comprehensive surveillance system designed to identify epidemic-prone syndromes and enable rapid public health interventions. Methods We analysed data from the Senegalese 4S real-time sentinel syndromic surveillance network collected between 2015 (15 sites) and 2023 (27 sites). The network monitored four key febrile syndromes, including malaria, dengue-like syndromes, diarrheal syndrome, and influenza-like illness (ILI), via standardized WHO case definitions. Laboratory confirmation was achieved through molecular and serological testing of biological samples. Sentinel general practitioners submitted daily reports via a digital platform that facilitated real-time reporting and automated alert generation. We evaluated system performance through completeness, timeliness, temporal patterns, geographical distribution, and alert validation rates. Results During the nine-year surveillance period, the network documented 1,816,340 outpatient consultations, with febrile syndromes accounting for 11.7% of all visits and demonstrating notable annual fluctuations. Distinct regional patterns of infectious disease events were observed: ILI predominated in western regions, dengue-like syndromes were clustered in north-central areas, and malaria cases were concentrated in southeastern zones. The system demonstrated robust performance metrics, achieving 94.5% data completeness and 80.0% reporting timeliness. Of the 202 alerts generated, 51.0% received laboratory confirmation. Dengue virus circulation was documented in 2017, 2018, 2021, 2022, and 2023. Despite these successes, 37.1% of febrile cases remained etiologically unclassified. The system's early multidisciplinary investigation capabilities enabled swift outbreak containment and transmission control. Conclusion The 4S network validates the effectiveness and practical implementation of digital, real-time syndromic surveillance in Senegal. It successfully facilitated early outbreak detection and supported prompt public health responses. Although the system has significant potential for resource-constrained environments, addressing current operational limitations remains crucial for maximizing public health impacts. These findings provide strong evidence supporting the regional expansion of similar surveillance frameworks to enhance health security and epidemic preparedness throughout West Africa.

Accurate differentiation between viral and bacterial infections remains challenging in resource-limited, arbovirus-endemic settings, leading to antibiotic misuse and diagnostic uncertainty. Myxovirus resistance protein A (MxA), an interferon-induced host biomarker, may offer a pathogen-agnostic approach to improve rapid diagnosis and clinical triage. We evaluated the performance of an MxA rapid diagnostic test (RDT) using archived samples from febrile patients collected during dengue virus (DENV) and chikungunya virus (CHIKV) outbreaks in Senegal. We tested 171 blood samples from patients with acute febrile illness using an MxA RDT and RT-qPCR for DENV and CHIKV. Samples with discordant results (MxA-positive and RT-qPCR-negative) underwent metagenomic and hybrid-capture Illumina-based sequencing to detect missed infections. Sequencing data were analyzed using maximum-likelihood phylogenetics to assess viral lineage placement. The MxA RDT demonstrated moderate-to-high sensitivity (70.0%-85.1%, depending on virus) and moderate specificity (70.2%) for detecting primary arboviral infections. Among discordant samples, sequencing revealed previously missed pathogens, including DENV serotype 3 (genotype III), Parvovirus B19 (B19V), and Torque teno virus (TTV). Detection of B19V and TTV highlights the broader clinical utility of host-response biomarkers to uncover unexpected viral pathogens in high-diversity settings. MxA's longer persistence than viral RNA enables detection of recent infections missed by PCR. Combined with sequencing, this broadens the diagnostic window, improves clinical triage, and supports identification of underdiagnosed viruses. Future research should integrate MxA testing into routine clinical care and surveillance protocols to enhance outbreak responses in resource-limited regions.IMPORTANCETimely and equitable viral diagnosis is vital in outbreak-prone regions where advanced laboratories are scarce. This study shows how a simple, rapid test for the host biomarker myxovirus resistance protein A can provide real-time detection of viral infections such as dengue and chikungunya, even in remote or frontline health centers. When paired with pathogen sequencing, the test also uncovers infections that standard PCR may miss. This integrated approach demonstrates how field-deployable diagnostics can operate both during and between epidemics, strengthening outbreak preparedness, improving patient triage, and advancing laboratory equity worldwide.

Zika virus (ZIKV) is a mosquito-borne orthoflavivirus primarily transmitted among humans by Aedes aegypti. Over the past two decades, it has caused significant outbreaks associated with birth defects and neurological disorders. ZIKV consists of two main genotypes: the African and Asian lineages, each exhibiting distinct biological properties. African lineage strains are transmitted more efficiently by mosquitoes, but the genetic basis for this difference has been elusive. Here, we investigate this question by comparing recent African and Asian strains using chimeric viruses with swapped genome segments. Our results show that structural genes from the African strain enhance viral internalization, while non-structural genes improve genome replication and infectious particle production in mosquito cells. In vivo mosquito transmission is most significantly influenced by structural genes, although no single viral gene alone is decisive. We also develop a stochastic model of in vivo viral dynamics that reflects the observed patterns, suggesting the key difference between African and Asian strains lies in their ability to traverse mosquito salivary glands. Our findings imply the polygenic nature of ZIKV transmissibility has hindered Asian strains from achieving the same transmission efficiency as African strains, highlighting the role of lineage-specific adaptive landscapes in ZIKV evolution and emergence.

Crimean-Congo hemorrhagic fever (CCHF) is a zoonosis transmitted mainly by ticks. In Senegal, the CCHF virus has been circulating since at least 2003. However, the first symptomatic human case was detected and confirmed only in 2023 in the southeastern part of the country. Following this detection, a survey of humans, animals and ticks was launched to better characterize the ecology of this virus in this area. Human samples were tested by ELISA and RT-PCR. Serum and tick samples collected from cattle, sheep and goats were tested by ELISA and by RT-PCR, respectively. Antibodies against CCHFV were detected in 0.77 % of humans and 14.18 % of animal sera. The highest seroprevalence was observed in cattle (58.3 %), followed by goats (33.33 %) and sheep (8.33 %). The tick infestation rate was higher in cattle (8.2 %) than in sheep (1.2 %), while goats were not infested. Hyalomma rufipes was the first tick found infected in southeastern Senegal with a CCHF viral minimum field infection rate of 13.3 %. Thus, this study enabled us to better understand the mechanisms of CCHFV emergence in southeastern Senegal.

Background:Accurate and timely differential diagnoses are a challenge for health care, particularly in infrastructure-poor settings. Methods:To investigate fevers of unknown origin in Africa, a mobile suitcase laboratory was deployed to DRC and Southwest Nigeria to support the control of the 2018-2020 Ebola virus disease outbreak in North-Kivu and Ituri provinces (DRC) and to provide a point-of-need solution for malaria confirmation during the dry season, respectively. Results:In DRC, the samples were tested for Ebola virus and the differentials Plasmodium falciparum, Salmonella enterica, yellow fever virus, Dengue virus, and chikungunya virus. In Southwest Nigeria, the samples were not tested for Ebola virus but were tested for the same differentials and additionally for Rickettsia spp., Leptospira, and Streptococcus pneumoniae. Plasmodium falciparum was detected in 23% (n = 192) and 47% (n = 88) of cases, respectively, and Salmonella enterica was detected in only 1 case in each cohort. Conclusions:The etiological agents circulating in febrile patients in Sub-Saharan Africa and the true incidence of neglected tropical diseases are still underestimated.

Senegal has experienced periodic epidemics of dengue in urban areas with increased incidence in recent years. However, few data are available on the local ecology of the epidemic vectors. In October 2021, a dengue outbreak was reported in northern Senegal to the Institute Pasteur de Dakar. Entomologic investigations then were undertaken to identify the areas at risk of transmission and to identify the vector(s). Adult mosquitoes were collected indoors and outdoors at selected households, while containers with water were inspected for mosquito larvae. All the Aedes aegypti (L.) collected were tested for dengue virus NS1 protein using a rapid diagnostic test (RDT), and positive samples were confirmed by real-time RT-PCR. The qRT-PCR positive samples were subjected to whole genome sequencing using Nanopore technology. The majority of the larvae-positive containers (83.1%) were used for water storage. The Breteau and Container indices exceeded the WHO-recommended thresholds for the risk of dengue virus transmission except at 2 localities. Ae. aegypti, the only reputed dengue vector, was collected resting indoors as well as outdoors and biting during the day and night. The NS1 protein was detected in 22 mosquito pools, including one pool of females emerging from field-collected larvae. All NS1-positive results were confirmed by RT-PCR. Virus serotyping showed that the outbreak was caused by DENV-1. This study demonstrates the need for continuous control of adult and aquatic stages of Ae. aegypti to prevent future dengue epidemics in Senegal. RDTs appear to be a promising tool for dengue diagnostics and surveillance.

Rapid proliferation of traditional gold mining sites in Kédougou a Southeast region in Senegal, led to mass population migration from the neighboring West African countries and rapid expansion of small mining villages with poor hygiene and sanitation conditions. An outbreak of hepatitis E was reported in 2014 with several cases of febrile jaundice among traditional mine workers. In this study, we analyzed both HEV IgM and IgG seroprevalence and the associated risk factors of infection by testing any suspected case and contacts collected from February 2012 to November 2014. RNA-negative sera from suspected cases and contacts were tested for anti-HEV IgM and anti-HEV IgG. A total of 799 sera were collected from 290 suspected cases, 470 contacts and 39 individuals with missing information. The median age of the cohort study was 19 years (1-88 years) with a male/female sex-ratio of 1.9. We found an overall prevalence of 43.68% (332/760) of anti-HEV IgM and 38.15% (290/760) of anti-HEV IgG sera. Our data provide new insights into the HEV epidemiology and point to the crucial need to estimate the disease’s burden in Kédougou and assess the viral mechanisms driving the disease’s severity in pregnant women.

Management of the COVID-19 pandemic relies on molecular diagnostic methods supported by serological tools. Herein, we developed S-RBD- and N- based ELISA assays useful for infection rate surveillance as well as the follow-up of acquired protective immunity against SARS-CoV-2. ELISA assays were optimized using COVID-19 Tunisian patients' sera and prepandemic controls. Assays were further validated in 3 African countries with variable endemic settings. The receiver operating curve was used to evaluate the assay performances. The N- and S-RBD-based ELISA assays performances, in Tunisia, were very high (AUC: 0.966 and 0.98, respectively, p < 0.0001). Cross-validation analysis showed similar performances in different settings. Cross-reactivity, with malaria infection, against viral antigens, was noticed. In head-to-head comparisons with different commercial assays, the developed assays showed high agreement. This study demonstrates, the added value of the developed serological assays in low-income countries, particularly in ethnically diverse populations with variable exposure to local endemic infectious diseases.

In Senegal, the burden of dengue is increasing and expanding. As case management and traditional diagnostic techniques can be difficult to implement, rapid diagnostic tests (RDTs) deployed at point of care are ideal for investigating active outbreaks. The aim of this study was to evaluate the diagnostic performance of the Dengue NS1 and Dengue IgM/IgG RDTs on the serum/plasma samples in a laboratory setting and in the field. During laboratory evaluation, performance of the NS1 RDT was assessed using NS1 ELISA as the gold standard. Sensitivity and specificity were 88% [75–95%] and 100% [97–100%], respectively. Performance of the IgM/IG RDT was assessed using the IgM Antibody Capture (MAC) ELISA, indirect IgG, and PRNT as gold standards. The IgM and IgG test lines respectively displayed sensitivities of 94% [83–99%] and 70% [59–79%] and specificities of 91% [84–95%] and 91% [79–98%]. In the field, the Dengue NS1 RDT sensitivity and specificity was 82% [60–95%] and 75% [53–90%], respectively. The IgM and IgG test lines displayed sensitivities of 86% [42–100%] and 78% [64–88%], specificities of 85% [76–92%] and 55% [36–73%], respectively. These results demonstrate that RDTs are ideal for use in a context of high prevalence or outbreak setting and can be implemented in the absence of a confirmatory test for acute and convalescent patients.

Arthropod-borne diseases currently constitute a source of major health concerns worldwide. They account for about 50% of global infectious diseases and cause nearly 700,000 deaths every year. Their rapid increase and spread constitute a huge challenge for public health, highlighting the need for early detection during epidemics, to curtail the virus spread, and to enhance outbreak management. Here, we compared a standard quantitative polymerase chain reaction (RT-qPCR) and a direct RT-qPCR assay for the detection of Zika (ZIKV), Chikungunya (CHIKV), and Rift Valley Fever (RVFV) viruses from experimentally infected-mosquitoes. The direct RT-qPCR could be completed within 1.5 h and required 1 µL of viral supernatant from homogenized mosquito body pools. Results showed that the direct RT-qPCR can detect 85.71%, 89%, and 100% of CHIKV, RVFV, and ZIKV samples by direct amplifications compared to the standard method. The use of 1:10 diluted supernatant is suggested for CHIKV and RVFV direct RT-qPCR. Despite a slight drop in sensitivity for direct PCR, our technique is more affordable, less time-consuming, and provides a better option for qualitative field diagnosis during outbreak management. It represents an alternative when extraction and purification steps are not possible because of insufficient sample volume or biosecurity issues.

Zika virus (ZIKV) diagnostics are crucial for proper antenatal and postnatal care and also for surveillance and serosurvey studies. Since the viremia during ZIKV infection is fleeting, serological testing is highly valuable to inform diagnosis. However, current serology tests using whole virus antigens frequently suffer from cross reactivity issues, delays, and technical complexity, especially in low and middle income countries (LMICs) and endemic countries. Here, we describe an indirect ELISA to detect specific IgG antibodies using the ZIKV envelope domain III (EDIII) protein expressed in Drosophila S2 cells as an immunogen. Using a total of 367 clinical samples, we showed that the EDIII-ELISA was able to detect IgG antibodies against ZIKV with high sensitivity of 100.0% and specificity of 94.7% when compared to plaque reduction neutralization tests (PRNTs) as the gold standard and using 0.208 as the cut-off OD value. These results show the usefulness of the recombinant envelope domain III as an alternative to standard whole virus proteins for ZIKV diagnostics as it improves the sensitivity and specificity of IgG ELISA assay when used as an immunogen. This method should, therefore, be extended to serological diagnostic techniques for other members of the flavivirus genus and for use in IgM diagnostic testing.

On 10 th September 2018, the Syndromic Sentinel Surveillance network that monitors febrile illnesses in all 14 regions of Senegal detected a peak of fever in the Fatick region. On 13 September 2018, 10 samples were sent to the WHO Collaborating Centre for Arboviruses and Viral Haemorrhagic Fevers at the Institut Pasteur de Dakar (IPD). Laboratory investigations revealed an epidemic of dengue 1 genotype V and dengue 3 genotype III. Fatick neighbors the Holy City of Touba where 3.5 million people from all over the word gather every year for the Grand Magal pilgrimage. This article discusses the impact of mass gatherings and their role in the recent introduction of dengue serotypes in Senegal. Dengue is now endemic in Senegal and across many countries in Africa, highlighting the need for early detection, control measures and prevention of severe dengue cases in highly connected urban settings.

The genus Flavivirus in the Flaviridae contains arthropod born viruses associated with high public health burdens like Zika, Dengue or Yellow fever. Saboya virus (SABV) is an understudied flavivirus grouping in the same genetic sub-group as Yellow Fever Virus (YFV) together with Sepik virus (SEPV) and Wesselbron virus (WSLV). Flavivirus infections are characterized by non-specific clinical presentations resulting in a high risk of misdiagnosis. SABV virus has been shown to circulate in the Sahelian zone and in central Africa. To study this virus we a qRT-PCR system based on TaqMan chemistry was developed to allow rapid and specific detection of SABV. The SABV assay was evaluated on available SABV isolates and others flaviviruses (DENV, ZIKV, YFV, WNV, KEDV). The system reliably detected all used SABV strains without cross amplification of other flaviviruses. In term of sensitivity the SABV assay detect up to 40.25 copies of SABV standard DNA molecule per ul. This system can be easily added to the available panel of arboviruses detection assays as a reliable tool to study virus prevalence in human, vertebrate and insect-vector samples.

The Rapid proliferation of traditional gold mining sites in the Kedougou region has led to massive migration of people from neighbouring West African countries and the establishment of several small villages where poor hygiene and sanitation conditions exist. In this context, a Hepatitis E virus outbreak was reported in Kedougou in 2014 with several cases among the traditional mining workers. Herein, we described epidemiological and laboratory data collected during the outbreak’s investigation from February 2012 to November 2014. Any suspected, contact or probable case was investigated, clinical and epidemiological data were collected. In our study, sera were collected and tested for viral RNA and anti-Hepatitis E virus (HEV) IgM. Archived serum samples from Kedougou were retrospectively screened by real-time polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). A total of 65 water samples collected from ponds and wells surrounding gold panners' sites and habitats and 75 tissues samples from rats captured in the environment of traditional gold mining sites were also tested. A total of 1617 sera were collected from 698 suspected cases, 862 contacts and 57 persons with missing information. The median age was 20 (1–88 years-old) and the sex ratio was 1.72. An overall rate of 64.62% (1045/1617) of these patients tested positive for HEV with a high case fatality rate in pregnant women. All water samples and animal tissues tested negative for HEV. Our data help not only determining of the beginning of the HEV outbreak to March 2012, but also identifying risk factors associated to its emergence. However, there is a need to implement routine diagnosis, surveillance and training of health personnel in order to reduce mortality especially among pregnant women. In addition, further studies are needed to identify the virus reservoir and environmental risk factors for HEV in the Kedougou region.

Objectives: A nationwide cross-sectional epidemiological survey was conducted to capture the true extent of coronavirus disease 2019 (COVID-19) exposure in Senegal. Methods: Multi-stage random cluster sampling of households was performed between October and November 2020, at the end of the first wave of COVID-19 transmission. Anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies were screened using three distinct ELISA assays. Adjusted prevalence rates for the survey design were calculated for each test separately, and thereafter combined. Crude and adjusted prevalence rates based on test performance were estimated to assess the seroprevalence. As some samples were collected in high malaria endemic areas, the relationship between SARS-CoV-2 seroreactivity and antimalarial humoral immunity was also investigated. Results: Of the 1463 participants included in this study, 58.8% were female and 41.2% were male; their mean age was 29.2 years (range 0.20-84.8.0 years). The national seroprevalence was estimated at 28.4% (95% confidence interval 26.1-30.8%). There was substantial regional variability. All age groups were impacted, and the prevalence of SARS-CoV-2 was comparable in the symptomatic and asymptomatic groups. An estimated 4 744 392 (95% confidence interval 4 360 164-5 145 327) were potentially infected with SARS-CoV-2 in Senegal, while 16 089 COVID-19 RT-PCR laboratory-confirmed cases were reported by the national surveillance. No correlation was found between SARS-CoV-2 and Plasmodium seroreactivity. Conclusions: These results provide a better estimate of SARS-CoV-2 dissemination in the Senegalese population. Preventive and control measures need to be reinforced in the country and especially in the south border regions.

The explosive emergence of Zika virus (ZIKV) across the Pacific and Americas since 2007 was associated with hundreds of thousands of human cases and severe outcomes, including congenital microcephaly caused by ZIKV infection during pregnancy. Although ZIKV was first isolated in Uganda, Africa has so far been exempt from large-scale ZIKV epidemics, despite widespread susceptibility among African human populations. A possible explanation for this pattern is natural variation among populations of the primary vector of ZIKV, the mosquito Aedes aegypti. Globally invasive populations of Ae. aegypti outside of Africa are considered effective ZIKV vectors because they are human specialists with high intrinsic ZIKV susceptibility, whereas African populations of Ae. aegypti across the species' native range are predominantly generalists with low intrinsic ZIKV susceptibility, making them less likely to spread viruses in the human population. We test this idea by studying a notable exception to the patterns observed across most of Africa: Cape Verde experienced a large ZIKV outbreak in 2015 to 2016. We find that local Ae. aegypti in Cape Verde have substantial human-specialist ancestry, show a robust behavioral preference for human hosts, and exhibit increased susceptibility to ZIKV infection, consistent with a key role for variation among mosquito populations in ZIKV epidemiology. These findings suggest that similar human-specialist populations of Ae. aegypti in the nearby Sahel region of West Africa, which may be expanding in response to rapid urbanization, could serve as effective vectors for ZIKV in the future.

Purpose: West Nile virus (WNV) is an arthropod-borne zoonotic pathogen which represents a continuous source of concern for public health worldwide due to its expansion and invasion into new regions. Its distribution and circulation intensity in African countries is only partially known. The aim of the present study is to provide an updated overview on the current knowledge of WNV epidemiology in Africa, providing available data on incidence in humans and animals, the circulating lineages and clades, other than an updated list of the principal arthropod vectors identified and the availability of vector competence studies. Methods & Materials: We searched pertinent articles to be included in the Scoping Review according to PRISMA and QUORUM criteria. We searched PubMed and Google Scholar electronic databases on January 21, 2020, using selected keywords. From the references of each article, we explored further references as appropriate. Additional references have been later identified and added accordingly to expert opinion. Results: This review, based on the analysis of 150 scientific papers published between 1940 and 2020, highlights: (i) the co-circulation of WNV-lineages 1,2, and 8 in the African continent; (ii) the circulation of Koutango virus in Senegal, Gabon, Somalia, and Niger (iii) the presence of diverse WNV competent vectors in Africa, mainly belonging to the Culex genus; (iv) the lack of vector competence studies for several other mosquito species found naturally infected with WNV in Africa; (v) evidence of circulation of WNV among humans, animals and vectors in at least 28 Countries; and (vi) the lack of knowledge on the epidemiological situation of WNV for 19 Countries. Conclusion: This study provides the state of art on WNV investigation carried out in Africa, highlighting several knowledge gaps regarding i) the partial knowledge on the current WNV distribution and genetic diversity, ii) its ecology and transmission chains including the role of different arthropods and vertebrate species as competent reservoirs, and iii) the real disease burden for humans and animals, therefore highlighting the needs for further research and surveillance studies to be addressed with high priority in this Continent.This study was partially funded by EU grant 874850 MOOD.