Sandfly surveillance and investigation of Leishmania spp. DNA in sandflies in Kosovo.

In the past decade, leishmaniasis seems to be re-emerging in Balkan countries. There are serious implications that Kosovo is a visceral leishmaniasis endemic region with autochthonous transmission; nevertheless, surveillance of vectors, reservoirs or the disease is not yet established. Gaining knowledge about sandfly vector species is a prerequisite for the development of a monitoring and control plan in the future. After a long gap in research of over 70 years, sandfly studies in Kosovo were resumed in 2014. During this presence/absence study, nine sandfly species were detected: Phlebotomus papatasi, Ph. perfiliewi, Ph. tobbi, Ph. neglectus, Ph. simici, Ph. balcanicus, Ph. alexandri, Ph. mascittii and Sergentomyia minuta. Three species are new with regard to the fauna of Kosovo - Ph. alexandri, Ph. balcanicus and Ph. mascittii. Besides increased diversity, changes in the number of collected specimens and distribution range of species were recorded, with Ph. neglectus being the most dominant species with the widest distribution. Testing of randomly chosen females for Leishmania spp. DNA resulted the in detection of L. tropica in a specimen of Ph. neglectus. The presence of numerous vector species in the sandfly fauna of Kosovo pose a threat for the re-emergence of vector-borne diseases. Therefore, continuous surveillance is recommended with regular updates on vector distribution and abundance.

Risk factors of exposure to Aedes albopictus bites in mainland France using an immunological biomarker.

In recent decades, the invasive Aedes albopictus vector has spread across Europe and is responsible for numerous outbreaks of autochthonous arboviral disease. The aim of this study was to identify epidemiological and sociological risk factors related to individual levels of exposure to Aedes albopictus bites. A multidisciplinary survey was conducted with volunteer blood donors living in areas either colonised or not by Aedes albopictus in mainland France. Individual levels of exposure were evaluated by measuring the IgG level specific to Aedes albopictus saliva. The most striking risk factors concerned the localisation and characteristics of the dwelling. Individuals living in areas colonised prior to 2009 or recently colonised (between 2010 and 2012) had higher anti-salivary gland extract IgG levels compared with those who were living in areas not yet colonised by Ae. albopictus. The type of dwelling did not seem to impact the level of exposure to Aedes bites. People living in apartments had a higher anti-salivary gland extract IgG level than those living in individual houses but the difference was not statistically significant. Interestingly, the presence of air conditioning or window nets was associated with a noticeable reduction in bite intensity.

Japanese encephalitis in a French traveler to Nepal.

Japanese encephalitis is frequent in Asia, with a severe prognosis, but rare in travelers. Culex mosquitoes transmit Japanese encephalitis virus. Risk factors are destination, duration of stay, summer and fall seasons, outdoor activities, and type of accommodation. We report the case of a French traveler to Nepal with neutralization-based serological confirmed Japanese encephalitis. He presented classical clinical (viral syndrome before an encephalitis status with behavioral disorder, global hypotonia, mutism, movement disorders, seizure, and coma), radiological (lesions of thalami, cortico-spinal tracts, and brainstem) and biological features (lymphocytic meningitis). Nowadays, the presence of Japanese encephalitis virus in Nepal, including mountain areas, is established but Japanese encephalitis remains rare in travelers returning from this area and neurologist physicians need to become familiar with this. We recommend vaccination for travelers spending a long period of time in Nepal and having at-risk outdoor activities.

Arthropods as a source of new RNA viruses.

The discovery and development of methods for isolation, characterisation and taxonomy of viruses represents an important milestone in the study, treatment and control of virus diseases during the 20th century. Indeed, by the late-1950s, it was becoming common belief that most human and veterinary pathogenic viruses had been discovered. However, at that time, knowledge of the impact of improved commercial transportation, urbanisation and deforestation, on disease emergence, was in its infancy. From the late 1960s onwards viruses, such as hepatitis virus (A, B and C) hantavirus, HIV, Marburg virus, Ebola virus and many others began to emerge and it became apparent that the world was changing, at least in terms of virus epidemiology, largely due to the influence of anthropological activities. Subsequently, with the improvement of molecular biotechnologies, for amplification of viral RNA, genome sequencing and proteomic analysis the arsenal of available tools for virus discovery and genetic characterization opened up new and exciting possibilities for virological discovery. Many recently identified but "unclassified" viruses are now being allocated to existing genera or families based on whole genome sequencing, bioinformatic and phylogenetic analysis. New species, genera and families are also being created following the guidelines of the International Committee for the Taxonomy of Viruses. Many of these newly discovered viruses are vectored by arthropods (arboviruses) and possess an RNA genome. This brief review will focus largely on the discovery of new arthropod-borne viruses.

Afebrile meningoencephalitis with transient central facial paralysis due to Toscana virus infection, southeastern France, 2014 [corrected].

We report a case of meningoencephalitis caused by Toscana virus (TOSV) with central facial paralysis lasting over two days acquired in south-eastern France. The patient was not febrile either before or during the course of the disease. The diagnosis was established by both real-time RT-PCR and virus isolation with complete genome sequencing. This case emphasises the need to consider TOSV in non-febrile neurological syndromes in people living in or having travelled to the Mediterranean area.

Phlebotomine sandflies and the spreading of leishmaniases and other diseases of public health concern.

Phlebotomine sandflies transmit pathogens that affect humans and animals worldwide. We review the roles of phlebotomines in the spreading of leishmaniases, sandfly fever, summer meningitis, vesicular stomatitis, Chandipura virus encephalitis and Carrión's disease. Among over 800 species of sandfly recorded, 98 are proven or suspected vectors of human leishmaniases; these include 42 Phlebotomus species in the Old World and 56 Lutzomyia species in the New World (all: Diptera: Psychodidae). Based on incrimination criteria, we provide an updated list of proven or suspected vector species by endemic country where data are available. Increases in sandfly diffusion and density resulting from increases in breeding sites and blood sources, and the interruption of vector control activities contribute to the spreading of leishmaniasis in the settings of human migration, deforestation, urbanization and conflict. In addition, climatic changes can be expected to affect the density and dispersion of sandflies. Phlebovirus infections and diseases are present in large areas of the Old World, especially in the Mediterranean subregion, in which virus diversity has proven to be higher than initially suspected. Vesiculovirus diseases are important to livestock and humans in the southeastern U.S.A. and Latin America, and represent emerging human threats in parts of India. Carrión's disease, formerly restricted to regions of elevated altitude in Peru, Ecuador and Colombia, has shown recent expansion to non-endemic areas of the Amazon basin.

Outbreak of adenovirus D8 in a neonatal intensive care unit involving multiple simultaneous transmission pathways.

BACKGROUND: Adenovirus (ADV) outbreaks in neonatal intensive care units (NICU) can lead to durable transmission and serious adverse outcomes. This study describes the investigation and control of an ADV-D8 outbreak in an NICU, associated with ophthalmologic equipment used during retinopathy of prematurity (ROP) screening. Cases were observed in neonates, parents and nurses. METHODS: The outbreak investigation was performed including sampling patients, parents and health care workers as well as the environment for molecular detection of ADV DNA. The investigation was also conducted in the guest house where some parents were temporary residents. A retrospective cohort study focused on neonates hospitalized during the epidemic period to assess the risk associated with ROP examination. RESULTS: Fifteen cases were identified in neonates; all but one presented with conjunctivitis. Two healthcare workers and 18 parents acquired conjunctivitis. ADV DNA was identified on the RetCam and on the freezer shared by parents. All ADV-positive samples were typed as ADV-D8. ADV infections occurred more frequently in neonates who had ROP examinations (37.8% (14/37) vs (0.9% (1/110); P<0.001) (relative risk 41.6; (5.7-305.8)). The RetCam was disinfected between two examinations using a disinfectant that was virucidal on ADV after a 30-min contact. CONCLUSION: This outbreak was significantly associated with ROP examination with a RetCam that had a disinfection protocol ill-adapted to rapid patient turnover. In addition, nosocomial transmission via the parents to neonates and parent-to-parent transmission is likely to have played a role in the dissemination of cases. No further cases were observed after the new disinfection procedure was enforced.

The viral etiology of an influenza-like illness during the 2009 pandemic.

Many viruses are known to cause influenza-like illness (ILI); however, in nearly 50% of patients, the etiologic agent remains unknown. The distribution of viruses in patients with ILI was investigated during the 2009 A/H1N1 influenza pandemic (A/H1N1p). From June 2009 to January 2010, 660 patients with suspected influenza were questioned and examined, and nasal swabs were collected. All patient samples were tested for influenza virus, and 286 negative nasal swabs were tested further for 18 other respiratory viruses using real-time RT-PCR. Two waves of ILI were observed in the epidemic curve (weeks 35-42 and 42-49). At least eight viruses co-circulated during this period: human rhinovirus (HRV) (58), parainfluenza 1-4 viruses (PIV) (9), human Coronavirus (hCoV) OC43 (9), enterovirus (5), adenovirus (AdV) (4), and human metapneumovirus (hMPV) (2); however, 204 samples remained negative for all viruses tested. ILI symptoms, according to the Centers for Disease Control and Prevention criteria for ILI definition, were reported in 75% of cases. These patients had positive swabs for A/H1N1p, HRV, hCoV-OC43, PIV, AdV, and hMPV without significant difference with non-ILI patients. This study found that many respiratory viruses circulated during this period and that the A/H1N1p did not impact on the kinetics of other respiratory viruses. The proportion of non-documented cases remains high. ILI could not distinguish A/H1N1p infection from that due to other respiratory viruses. However, in multivariate anlaysis, cough, chills, hyperemia, and dyspnea were associated significantly with influenza virus versus other respiratory viruses.

Measles among healthcare workers: a potential for nosocomial outbreaks.

We report here 14 cases of measles among healthcare workers (HCWs) in Public Hospitals of Marseilles, France that occurred between April and November 2010. All cases but one were under 30 years of age. Following the identification of these cases, we checked the immune status among 154 HCWs who volunteered to take part in the study and showed that 93% and 88% were immune against measles and mumps respectively. HCWs non-immunised against measles were all under 30 years of age.

Record of Phlebotomus (Transphlebotomus) mascittii Grassi, 1908 and Phlebotomus (Larroussius) chadlii Rioux, Juminer & Gibily, 1966 female in Algeria.

We report for the first time the presence of Phlebotomus mascittii and the female of Phlebotomus chadlii in Algeria. These two species were collected during an entomological study conducted in endemic visceral leishmaniasis focus from the north part of the country, Kabylia.

Evaluation of formulations to improve SARS-CoV-2 viability and thermostability after lyophilisation.

In the context of the COVID-19 pandemic, virus collections such as EVA-GLOBAL play a key role in the supply of viruses and related products for research. Freeze-drying techniques for viruses represent a method of choice for the preservation of strains and their distribution without the need for a demanding cold chain. Here, we describe an optimised lyophilisation protocol usable for SARS-CoV-2 strains that improves preservation and thermostability. We show that sucrose used as an adjuvant represents a simple and efficient stabilizer providing increased protection for long-term preservation and shipment of the virus under different climatic conditions.

Dengue virus infections in travellers returning from Benin to France, July-August 2010.

In July and August 2010, two cases of dengue fever were diagnosed in travellers returning from Benin to France. These two cases exemplify that dengue fever should be considered in febrile travellers, even those returning from areas where the infection is not usual.

An update on Toscana virus distribution, genetics, medical and diagnostic aspects.

BACKGROUND: Toscana virus is an arbovirus transmitted by sand flies within the Mediterranean area where it can cause febrile illness and neuroinvasive infections during the seasonal circulation period of the vector. Although it is an important cause of meningitis and encephalitis, it remains a neglected virus with limited published data, as demonstrated by <250 peer-reviewed articles since the 1970s. OBJECTIVE: The last review article on Toscana virus was published in 2012. The aim was to compile peer-reviewed articles to provide an updated review highlighting recent findings to complement previous review articles. SOURCES: PubMed database was searched using the 'Toscana virus' keyword from 2010 to present. A total of 152 articles were retrieved and identified studies were assessed for novel information on virus genetics, and geographic and medical aspects compared with existing knowledge reported in previous review articles. CONTENT: Studies addressing medical, veterinary and entomological aspects have provided evidence that Toscana virus is present in North Africa, in the Balkan Peninsula, and in most of the Mediterranean islands. Besides the two previously recognized genetic lineages, a novel evolutionary lineage has been identified in the Balkan Peninsula. Co-circulation of two genetic lineages has been demonstrated in France, in Turkey and in Croatia. In addition to meningitis and meningo-encephalitis, which have been reported for 40 years, various neuroinvasive forms have been recently reported such as Guillain-Barré syndrome, hydrocephalus, myositis, fasciitis, polymyeloradiculopathy, deafness and facial paralysis. IMPLICATION: Because it is endemic in countries bordering the Mediterranean, physicians should include Toscana virus in the differential diagnosis of patients presenting with febrile illness and/or neurological manifestations.

Molecular investigation of tick-borne pathogens in ixodid ticks infesting domestic animals (cattle and sheep) and small rodents (black rats) of Corsica, France.

Although livestock farming (sheep, goats, pigs, and cattle) is an important economic activity in Corsica, a French Mediterranean island, knowledge about the tick fauna and microorganisms carried by them remains scarce. This study aimed to investigate the presence and perform molecular characterization of Anaplasmataceae, Rickettsia spp., and Borrelia burgdorferi sensu lato (sl) in tick species collected in Corsica. Ticks from cattle (Bos taurus), sheep (Ovis aries), and rodents (Rattus rattus) were collected from May to September 2016. DNA was purified from ticks, submitted to quantitative real-time polymerase chain reaction (qPCR) and sequenced for phylogenetic analysis. In total, 660 ticks were collected from 111 animals during the study. The most abundant collected tick species from cattle was Rhipicephalus bursa (n = 495; 84.5%), followed by Hyalomma marginatum (n = 91; 15.5%). Rhipicephalus bursa and Ixodes ricinus were the only tick species collected from sheep and rodents, respectively. Overall, Rickettsia was the most common pathogen group (n = 48; 24%) detected in ticks. Sequence analysis of partial gltA and ompA genes revealed the presence of Ri. aeschlimannii and Candidatus Ri. barbariae. Anaplasmataceae DNA was detected in eight (6%) of the 127 cattle pools and in one (2%) of the 61 R. bursa specimens collected from sheep. Sequence analysis of the rpoB gene revealed the presence of one Anaplasma species, A. marginale. Borrelia burgdorferi sl DNA was detected in one pool of H. marginatum collected from cattle and in two (15%) of the 13 I. ricinus pools collected from nine black rats. To our knowledge, this is the first report of the occurrence and molecular characterization of Candidatus Ri. barbariae, an emerging member of the Rickettsia group causing spotted fever, in Corsica. The detection of B. burgdorferi sl DNA, which was previously believed to be rare in Corsica, confirms the presence of this agent on the island.

Arenaviruses.

The Arenaviridae family contains 22 recognized virus species, each of them strongly associated with a rodent species (except Tacaribe virus which is associated with a species of bat), suggesting an ancient co-evolutionary process. Although the concept of co-evolution between rodents and arenaviruses is now largely accepted, little has been uncovered in terms of dating the phenomenon and the mechanisms of evolution, including speciation and pathogenicity. These questions are targeted in the present chapter. Old World arenaviruses are associated with the Eurasian rodents in the family Muridae. New World arenaviruses are associated with American rodents in the subfamily Sigmodontinae. The correlation between the rodent host phylogeny and the viruses suggests a long association and a co-evolutionary process. Furthermore, three distinct New World arenaviruses share a common ancestor, demonstrating a unique recombination event that probably occurred in that ancestor. This shows that recombination among arenaviruses of different lineages might occur in nature. Recombination and co-evolutionary adaptation appear as the main mechanisms of arenavirus evolution, generating a high degree of diversity. The diversity among rodent host reservoir and virus species and the potential to exchange genomic material provide a basis for the emergence of new viruses and the risk of these becoming pathogenic for humans.

The VIZIER project: preparedness against pathogenic RNA viruses.

Life-threatening RNA viruses emerge regularly, and often in an unpredictable manner. Yet, the very few drugs available against known RNA viruses have sometimes required decades of research for development. Can we generate preparedness for outbreaks of the, as yet, unknown viruses? The VIZIER (VIral enZymes InvolvEd in Replication) (http://www.vizier-europe.org/) project has been set-up to develop the scientific foundations for countering this challenge to society. VIZIER studies the most conserved viral enzymes (that of the replication machinery, or replicases) that constitute attractive targets for drug-design. The aim of VIZIER is to determine as many replicase crystal structures as possible from a carefully selected list of viruses in order to comprehensively cover the diversity of the RNA virus universe, and generate critical knowledge that could be efficiently utilized to jump-start research on any emerging RNA virus. VIZIER is a multidisciplinary project involving (i) bioinformatics to define functional domains, (ii) viral genomics to increase the number of characterized viral genomes and prepare defined targets, (iii) proteomics to express, purify, and characterize targets, (iv) structural biology to solve their crystal structures, and (v) pre-lead discovery to propose active scaffolds of antiviral molecules.

Prospective comparative study of characteristics associated with influenza A and B in adults.

OBJECTIVES: To describe and compare the epidemiological characteristics associated with influenza type A and B as well as the characteristics associated with influenza pneumonia. The secondary objective was to evaluate the performance of influenza rapid diagnostic tests (RDT) in the emergency department. PATIENTS AND METHODS: Prospective study, including 251 adult patients admitted to the emergency department during the 2013-2014 influenza outbreaks for flu-like illness confirmed by RT-PCR. RESULTS: A total of 106 patients were infected with influenza type B (42%) and 145 with influenza type A (58%). Mean age was 56 and the sex-ratio was 0.9. In a multivariate analysis, the only factor independently related with the type of influenza strain was the patient's age. Overall, 17% of influenza-infected patients were vaccinated and 38% presented with pneumonia, with no significant difference between strains. In a univariate analysis, the risk factors associated with the occurrence of pneumonia were age, vaccination, and chronic obstructive pulmonary disease. Only 22% of samples positive by RT-PCR for influenza B and 40% for influenza A were positive by RDT. CONCLUSION: Influenza type A and type B had similar clinical and biological signs, including severity. Influenza type B should not be neglected. Any emergency department may use the RDT for its ease-of-use and rapidity. However, its low sensitivity should be taken into consideration when interpreting results.

Neglected vector-borne bacterial diseases and arboviruses in the Mediterranean area.

Arthropod vectors can transmit pathogenic microorganisms from one vertebrate to another during their blood meal. Although some vector-borne diseases have been eradicated in the Mediterranean area, such as malaria and dengue, recent endemic microorganisms (Toscana virus, Rickettsia spp.) remain neglected even though they cause many more cases. New diagnostic tools and innovative tools for the identification and characterization of vector species and microorganisms have been developed at IHU Méditerranée Infection, either internally or through collaborative and integrated projects. We have detected Rickettsia slovaca as a human pathogen and have described the disease; we have shown that Rickettsia felis can be transmitted by Anopheles mosquitoes; we have emphasized the increasing importance of bedbug (Cimex lectularius) as a potential vector of Bartonella quintana; and we have described the Toscana virus, a major agent of meningitis and meningoencephalitis which was disseminated in North Africa and Central and Eastern Europe, where it frequently cocirculates with a large number of newly described phleboviruses transmitted by sand flies.

First complete genome sequence of circulating dengue virus serotype 3 in Jeddah, Saudi Arabia.

Here we report the first full-length genome sequence of dengue virus serotype 3 (DENV-3) from a strain isolated from a patient in Jeddah, Saudi Arabia, in 2014. The genome consists of 10 635 bp and shows close similarity to circulating genotype III isolates from Singapore, suggesting possible importation, most probably during religious pilgrimages to Saudi Arabia.