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In 2019, a new coronavirus (2019-nCoV) infecting Humans has emerged in Wuhan, China. Its genome has been sequenced and the genomic information promptly released. Despite a high similarity with the genome sequence of SARS-CoV and SARS-like CoVs, we identified a peculiar furin-like cleavage site in the Spike protein of the 2019-nCoV, lacking in the other SARS-like CoVs. In this article, we discuss the possible functional consequences of this cleavage site in the viral cycle, pathogenicity and its potential implication in the development of antivirals., Highlights • The genomic sequence of 2019-nCoV indicates that the virus clusters with betacoronaviruses of lineage b. • 2019-nCoV S-protein sequence has a specific furin-like cleavage site absent in lineage b CoV including SARS-CoV sequences. • The furin-like cleavage site in the S-protein of 2019-nCoV may have implications for the viral life cycle and pathogenicity. • Campaigns to develop anti-2019-nCoV therapeutics should include the evaluation of furin inhibitors.

The GloPID-R (Global Research Collaboration for Infectious Disease Preparedness) chikungunya (CHIKV), o'nyong-nyong (ONNV) and Mayaro virus (MAYV) Working Group has been established to investigate natural history, epidemiology and clinical aspects of infection by these viruses. Here, we present a report dedicated to entomological aspects of CHIKV, ONNV and MAYV. Recent global expansion of chikungunya virus has been possible because CHIKV established a transmission cycle in urban settings using anthropophilic vectors such as Aedes albopictus and Aedes aegypti. MAYV and ONNV have a more limited geographic distribution, being confined to Africa (ONNV) and central-southern America (MAYV). ONNV is probably maintained through an enzootic cycle that has not been characterized yet, with Anopheles species as main vectors and humans as amplification hosts during epidemics. MAYV is transmitted by Haemagogus species in an enzootic cycle using non-human primates as the main amplification and maintenance hosts, and humans becoming sporadically infected when venturing in or nearby forest habitats. Here, we focused on the transmission cycle and natural vectors that sustain circulation of these viruses in their respective locations. The knowledge of the natural ecology of transmission and the capacity of different vectors to transmit these viruses is crucial to understand CHIKV emergence, and to assess the risk that MAYV and ONNV will expand on wide scale using anthropophilic mosquito species not normally considered primary vectors. Finally, the experts identified knowledge gaps and provided adapted recommendations, in order to address future entomological investigations in the right direction. ispartof: ANTIVIRAL RESEARCH vol:174 ispartof: location:Netherlands status: published

The GloPID-R (Global Research Collaboration for Infectious Disease Preparedness) Chikungunya (CHIKV), O'nyong-nyong (ONNV) and Mayaro virus (MAYV) Working Group is investigating the natural history, epidemiology and medical management of infection by these viruses, to identify knowledge gaps and to propose recommendations for direct future investigations and rectification measures. Here, we present the first report dedicated to diagnostic aspects of CHIKV, ONNV and MAYV. Regarding diagnosis of the disease at the acute phase, molecular assays previously described for the three viruses require further evaluation, standardized protocols and the availability of international standards representing the genetic diversity of the viruses. Detection of specific IgM would benefit from further investigations to clarify the extent of cross-reactivity among the three viruses, the sensitivity of the assays, and the possible interfering role of cryoglobulinaemia. Implementation of reference panels and external quality assessments for both molecular and serological assays is necessary. Regarding sero-epidemiological studies, there is no reported high-throughput assay that can distinguish among these different viruses in areas of potential co-circulation. New specific tools and/or improved standardized protocols are needed to enable large-scale epidemiological studies of public health relevance to be performed. Considering the high risk of future CHIKV, MAYV and ONNV outbreaks, the Working Group recommends that a major investigation should be initiated to fill the existing diagnostic gaps. ispartof: ANTIVIRAL RESEARCH vol:166 pages:66-81 ispartof: location:Netherlands status: published

Background Japanese encephalitis virus (JEV) is a leading cause of central nervous system (CNS) infections in Asia and results in significant morbidity and mortality. JEV RNA is rarely detected in serum or cerebrospinal fluid (CSF), and diagnosis of JEV infection is usually based on serological tests that are frequently difficult to interpret. Unlike serum or CSF, urine is relatively easy to obtain, but, to date, there has been minimal work on the feasibility of testing urine for JEV RNA. Methods We investigated the use of lysis buffer and a Microsep device to optimize urine storage for detection of JEV RNA by reverse transcription real-time polymerase chain reaction (RT-qPCR). The best of the studied methods was then evaluated in consecutive patients admitted to the hospital with suspected CNS infections in Laos. Results We demonstrated degradation of JEV RNA in urine after even short storage periods at 4°C or –80°C. Although there was no advantage in using a Microsep concentration device alone, immediate addition of lysis buffer to fresh urine improved the detection of JEV RNA at the limit of detection. Conclusions In 2 studies of 41 patients with acute encephalitis syndrome, 11 (27%) were positive for JEV IgM in CSF and/or serum, and 2 (4.9%) were JEV RT-qPCR positive from throat swabs. JEV RNA was not detected in any of these patients’ urine samples. However, lysis buffer was only used during a prospective study, that is, for only 17/41 (41%) patient urine samples. Our findings suggest a need for larger studies testing urine for JEV RNA, with urine collected at different times from symptom onset, and using lysis buffer, which stabilizes RNA, for storage.

Background Aedes albopictus, vector of dengue and chikungunya viruses, is implanted in mainland France, exposing to the risk of autochthonous transmission. Since 2006, epidemiological and entomological surveillance activities aim to prevent or limit the occurrence of autochthonous cases. We aimed to describe episodes of transmission and control measures implemented in order to reflect on surveillance activities. Methods We reviewed all publications and documents produced on autochthonous transmission episodes in France and surveillance protocols. We reviewed surveillance activities, investigation methods and control measures implemented. Results Between 2010 and 2018, eight episodes of autochthonous dengue fever transmission and three of chikungunya were recorded in mainland France. All of them occurred in the South east of France, between July and October, when vector density was the highest. Transmission areas were limited to single domestic houses located in discontinuous urban areas. Only two episodes happened in two distinct areas. Chikungunya episodes led to 31 cases and dengue fever episodes to 23 cases. Most cases were identified by door-to-door investigations set-up in transmission areas. We isolated serotypes 1 and 2 for dengue and East Central South Africa lineage for chikungunya in autochthonous cases. Adulticide vector control measures were effective in controlling transmission. Seven episodes of transmission were due to failure in identifying primary imported cases. Four episodes occurred because of the absence or the lack of vector controls measures around primary imported cases. Conclusions Surveillance activities, and autochthonous cases investigations, were effective in limiting the extent of transmission, but were highly demanding for surveillance actors. Identified causes of transmission highlight the need of regular awareness campaigns targeting physicians and biologists. Key messages Effectiveness of the surveillance system of dengue, chikungunya and zika viruses, and autochthonous cases investigations. Needs of awareness and training courses targeting health professionals to the risk represented by these viruses.

The European Virus Archive (EVA) was created in 2008 with funding from the FP7-EU Infrastructure Programme, in response to the need for a coordinated and readily accessible collection of viruses that could be made available to academia, public health organisations and industry. Within three years, it developed from a consortium of nine European laboratories to encompass associated partners in Africa, Russia, China, Turkey, Germany and Italy. In 2014, the H2020 Research and Innovation Framework Programme (INFRAS projects) provided support for the transformation of the EVA from a European to a global organization (EVAg). The EVAg now operates as a non-profit consortium, with 26 partners and 20 associated partners from 21 EU and non-EU countries. In this paper, we outline the structure, management and goals of the EVAg, to bring to the attention of researchers the wealth of products it can provide and to illustrate how end-users can gain access to these resources. Organisations or individuals who would like to be considered as contributors are invited to contact the EVAg coordinator, Jean-Louis Romette, at jean-louis.romette@univmed.fr., Highlights • The EVAg was created as an international organization aiming to provide a gold standard resource to the scientific community. • The EVAg operates as a non-profit consortium of 26 partners and 20 associated partners from EU and non-EU countries. • Members and associated members retain ownership of the viruses that they disseminate via the EVAg infrastructure. • The EVAg approach to quality management is directed by the project's own quality standard, based upon OECD guidelines. • The ultimate objective is to make the EVAg a permanent archive that can provide access to viruses and reagents globally.

International audience; Objectives: Cytomegalovirus (CMV) reactivation in intensive care unit patients may increase mortality and favour bacterial pneumonia. We developed a murine model to compare the severity of staphylococcal pneumonia after CMV reactivation and in CMV-negative mice. Methods: Balb/c mice were primo-infected with murine cytomegalovirus (MCMV n = 90) or received saline (control n = 90). After latency, all mice underwent caecal ligation and puncture to trigger MCMV reactivation in MCMV primary-infected mice. Surviving animals received an intra-nasal inoculation with methicillin-susceptible Staphylococcus aureus (MSSA) to induce pneumonia. Mortality, lung bacterial count, histology and interferon-alpha and gamma serum levels were compared in MCMV reactivated and control mice 2, 5 and 15 days after pneumonia. Results: AfterMSSA pneumonia, MCMV mice showed a trend towards a higher mortality (9.4% versus 0%; p 0.09) and a higher weight loss (2.2 (0.6-4.1 g) versus 0.7 (-0.3 to 1.3 g); p 0.005). The lung bacterial countwas higher in MCMV mice 2 days (5 x 10(3) (10(3) to 3 x 10(5)) versus 10(2) (0 to 4 x 10(2)) CFU/lung; p 0.007) and 5 days (2.5 x 10(4) (1.6 x 10(4) to 6.5 x 10(5)) versus 15 (10-40) CFU/lung; p 0.005) after MSSA pneumonia. 8/40 (20%) MCMV mice developed lung abscesses compared to 0% in control (p 0.011). Interferon-alpha serum levels 2 days after staphylococcal pneumonia were higher in MCMV mice. Conclusions: MCMV reactivation decreased lung bacterial clearance and favoured the development of staphylococcal abscessing pneumonia. CMV reactivation may be responsible for a higher susceptibility to bacterial sepsis. (C) 2016 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Highlights ► EVA is a globally available virus collection serving academia, public health and industry. ► EVA is a unique, networked, quality-controlled non-profit virus archive that benefits science. ► EVA provides wide-ranging access to virus collections held in laboratories worldwide. ► Laboratories in developing countries contribute to the pool of quality-controlled reagents., The European Virus Archive (EVA) was conceived as a direct response to the need for a coordinated and readily accessible collection of viruses that could be made available to academia, public health organisations and industry, initially within Europe, but ultimately throughout the world. Although scientists worldwide have accumulated virus collections since the early twentieth century, the quality of the collections and the viruses collected may vary according to the personal interests and agenda of the scientists. Moreover, when laboratories are re-organised or closed, collections are no longer maintained and gradually cease to exist. The tragedy of 9/11 and other disruptive activities have also meant that some previously available biological reagents are no longer openly exchanged between countries. In 2008, funding under the FP7–EU infrastructure programme enabled the initiation of the EVA. Within three years, it has developed from a consortium of nine European laboratories to encompass associated partners in Africa, Russia, China, Turkey, Germany and Italy. There is every reason to believe that EVA will continue to expand and ultimately exist as a globally networked, quality-controlled non-profit archive for the benefit of science. Organizations or individuals who would like to be considered as contributors are invited to contact the EVA coordinator, Jean–Louis Romette, at jean-louis.romette@univmed.fr.

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. J. Med. Virol. 84: 1071–1079, 2012. © 2012 Wiley Periodicals, Inc.

BACKGROUND: In the wake of the recent outbreak of Ebola virus disease (EVD) in several African countries, the World Health Organization prioritized the evaluation of treatment with convalescent plasma derived from patients who have recovered from the disease. We evaluated the safety and efficacy of convalescent plasma for the treatment of EVD in Guinea. METHODS: In this nonrandomized, comparative study, 99 patients of various ages (including pregnant women) with confirmed EVD received two consecutive transfusions of 200 to 250 ml of ABO-compatible convalescent plasma, with each unit of plasma obtained from a separate convalescent donor. The transfusions were initiated on the day of diagnosis or up to 2 days later. The level of neutralizing antibodies against Ebola virus in the plasma was unknown at the time of administration. The control group was 418 patients who had been treated at the same center during the previous 5 months. The primary outcome was the risk of death during the period from 3 to 16 days after diagnosis with adjustments for age and the baseline cycle-threshold value on polymerase-chain-reaction assay; patients who had died before day 3 were excluded. The clinically important difference was defined as an absolute reduction in mortality of 20 percentage points in the convalescent-plasma group as compared with the control group. RESULTS: A total of 84 patients who were treated with plasma were included in the primary analysis. At baseline, the convalescent-plasma group had slightly higher cycle-threshold values and a shorter duration of symptoms than did the control group, along with a higher frequency of eye redness and difficulty in swallowing. From day 3 to day 16 after diagnosis, the risk of death was 31% in the convalescent-plasma group and 38% in the control group (risk difference, -7 percentage points; 95% confidence interval [CI], -18 to 4). The difference was reduced after adjustment for age and cycle-threshold value (adjusted risk difference, -3 percentage points; 95% CI, -13 to 8). No serious adverse reactions associated with the use of convalescent plasma were observed. CONCLUSIONS: The transfusion of up to 500 ml of convalescent plasma with unknown levels of neutralizing antibodies in 84 patients with confirmed EVD was not associated with a significant improvement in survival. (Funded by the European Union's Horizon 2020 Research and Innovation Program and others; ClinicalTrials.gov number, NCT02342171.). ispartof: New England Journal of Medicine vol:374 issue:374 pages:33-42 ispartof: location:United States status: published

International audience; Human parvovirus B19 occurs worldwide and causes mild or asymptomatic disease in the form of cyclic local epidemics usually occurring in late winter and early summer. In 2012, a dramatic increase in cases was observed in the Public hospitals system of Marseille, with a total of 53 cases reported. Here, we describe the characteristics of this outbreak and compare it with the local epidemiology of B19V infections observed during the 2002–2011 period.

The swine-origin H1N1 influenza A virus (pH1N1(2009)) started to circulate worldwide in 2009, and cases were notified in a number of sub-Saharan African countries. However, no epidemiological data allowing estimation of the epidemic burden were available in this region, preventing comprehensive comparisons with other parts of the world. The CoPanFlu-Mali programme studied a cohort of 202 individuals living in the rural commune of Dioro (southern central Mali). Pre-pandemic and post-pandemic paired sera (sampled in 2006 and April 2010, respectively) were tested by the haemagglutination inhibition (HI) method. Different estimates of pH1N1(2009) infection during the 2009 first epidemic wave were used (increased prevalence of HI titre of ≥ 1/40 or ≥ 1/80, seroconversions) and provided convergent attack rate values (12.4-14.9%), the highest values being observed in the 0-19-year age group (16.0-18.4%). In all age groups, pre-pandemic HI titres of ≥1/40 were associated with complete absence of seroconversion; and geometric mean titres were 20 in others. Important variations in seroconversion rate existed among the different villages investigated. Despite limitations resulting from the size and composition of the sample analysed, this study provides strong evidence that the impact of the pH1N1(2009) first wave was more important than previously believed, and that the determinants of the epidemic spread in sub-Saharan populations were quite different from those observed in developed countries.

In May 2012, a Coxsackievirus A24 haemorrhagic conjunctivitis was diagnosed in Marseille, France, in a traveller returning from the Comoros Islands. This case allowed identification of the cause of an ongoing outbreak of haemorrhagic conjunctivitis in Indian Ocean Islands, illustrating that returning travellers may serve as sentinels for infectious diseases outbreaks in tropical areas where laboratory investigation is limited.

The 2C protein, which is an essential ATPase and one of the most conserved proteins across the Picornaviridae family, is an emerging antiviral target for which structural and functional characterization remain elusive. Based on a distant relationship to helicases of small DNA viruses, piconavirus 2C proteins have been predicted to unwind double-stranded RNAs. Here, a terminally extended variant of the 2C protein from echovirus 30 has been studied by means of enzymatic activity assays, transmission electron microscopy, atomic force microscopy and dynamic light scattering. The transmission electron-microscopy technique showed the existence of ring-shaped particles with similar to 12 nm external diameter. Image analysis revealed that these particles were hexameric and resembled those formed by superfamily 3 DNA virus helicases.

The alphaviruses were amongst the first arboviruses to be isolated, characterized and assigned a taxonomic status. They are globally very widespread, infecting a large variety of terrestrial animals, insects and even fish, and circulate both in the sylvatic and urban/peri-urban environment, causing considerable human morbidity and mortality. Nevertheless, despite their obvious importance as pathogens, there are currently no effective antiviral drugs with which to treat humans or animals infected by any of these viruses. The EU-supported project VIZIER (Comparative Structural Genomics of Viral Enzymes Involved in Replication, FP6 Project: 2004-511960) was instigated with an ultimate view of contributing to the development of antiviral therapies for RNA viruses, including the alphaviruses [Coutard, B., Gorbalenya, A.E., Snijder, E.J., Leontovich, A.M., Poupon, A., De Lamballerie, X., Charrel, R., Gould, E.A., Gunther, S., Norder, H., Klempa, B., Bourhy, H., Rohayemj, J., L'hermite, E., Nordlund, P., Stuart, D.I., Owens, R.J., Grimes, J.M., Tuckerm, PA., Bolognesi, M., Mattevi, A., Coll, M., Jones, TA., Aqvist, J., Unger, T., Hilgenfeld, R., Bricogne, G., Neyts, J., La Colla, P., Puerstinger, G., Gonzalez, J.P., Leroy, E., Cambillau, C., Romette, J.L., Canard, B., 2008. The VIZIER project: preparedness against pathogenic RNA viruses. Antiviral Res. 78, 37-46]. This review highlights some of the major features of alphaviruses that have been investigated during recent years. After describing their classification, epidemiology and evolutionary history and the expanding geographic distribution of Chikungunya virus, we review progress in understanding the structure and function of alphavirus replicative enzymes achieved under the VIZIER programme and the development of new disease control strategies. (C) 2009 Elsevier B.V. All rights reserved.

Some mammalian rhabdoviruses may infect humans, and also infect invertebrates, dogs, and bats, which may act as vectors transmitting viruses among different host species. The VIZIER programme, an EU-funded FP6 program, has characterized viruses that belong to the Vesiculovirus, Ephemerovirus and Lyssavirus genera of the Rhabdoviridae family to perform ground-breaking research on the identification of potential new drug targets against these RNA viruses through comprehensive structural characterization of the replicative machinery. The contribution of VIZIER programme was of several orders. First, it contributed substantially to research aimed at understanding the origin, evolution and diversity of rhabdoviruses. This diversity was then used to obtain further structural information on the proteins involved in replication. Two strategies were used to produce recombinant proteins by expression of both full length or domain constructs in either E. coil or insect cells, using the baculovirus system. In both cases, parallel cloning and expression screening at small-scale of multiple constructs based on different viruses including the addition of fusion tags, was key to the rapid generation of expression data. As a result, some progress has been made in the VIZIER programme towards dissecting the multi-functional L protein into components suitable for structural and functional studies. However, the phosphoprotein polymerase co-factor and the structural matrix protein, which play a number of roles during viral replication and drives viral assembly, have both proved much more amenable to structural biology. Applying the multi-construct/multi-virus approach central to protein production processes in VIZIER has yielded new structural information which may ultimately be exploitable in the derivation of novel ways of intervening in viral replication. (C) 2010 Elsevier B.V. All rights reserved.

To date, the International Committee for Taxonomy of Viruses recognizes that the family Arenaviridae contains a unique genus Arenavirus that includes 22 viral species. There are nine additional arenaviruses that either have been discovered recently, or which taxonomic status remains pending. Arenaviruses have been classified according to their antigenic properties into two groups, the Lassa-Lymphocytic choriomeningitis (LCM) serocomplex and the Tacaribe serocomplex which has been further divided into four evolutionary lineages. Each arenavirus is more or less tightly associated with a mammal host. The distribution of the host dictates the distribution of the virus. Humans may become infected by arenaviruses through direct contact with infected rodents, including bites, or through inhalation of infectious rodent excreta and secreta. Lassa, Junin, Machupo, Guanarito, and Sabia viruses are known to cause a severe hemorrhagic fever, in western Africa, Argentina, Bolivia, Venezuela, and Brazil, respectively. Infection by LCM virus can result in acute central nervous system disease, congenital malformations, and infection in organ transplantation recipients. Detection of arenaviruses in their animal host can be achieved by virus isolation, and has recently taken advantage of PCR-based techniques. The approach based on consensus degenerate primers has shown efficient for both detection of known arenaviruses, and discovery of new arenaviruses.

SUMMARYDuring the 2003 rainy season, a follow-up survey in sentinel chickens was undertaken to assess the seasonal transmission of West Nile virus (WNV) in a sahelian ecosystem: the Ferlo (Senegal). The estimated incidence rate in chickens was 14% (95% CI 7–29), with a very low level of transmission between July and September, and a transmission peak occurring between September and October. Comparing these results with the estimate obtained from a previous transversal serological study performed on horses the same year in the same area, the relevance of sentinel chickens in estimating the WNV transmission rate is highlighted. Conventionally adult mosquito populations disappear during the dry season but WN disease was shown to be endemic in the study area. The mechanisms of virus maintenance are discussed.

We demonstrate for the first time that a putative third member of the genus Anellovirus (TTV/TTMV) is present in the blood of healthy persons (20% prevalence), and also in their PBMNC and saliva samples.

Phlebotomine sandflies are vectors of a large variety of arboviruses, mostly in the genus Phlebovirus. Many were discovered during the last decade, but were only partially sequenced because of extremely high genetic diversity. Next-Generation sequencing (NGS) is a key tool to achieve rapid sequencing of these viruses. Here we present the results obtained with a custom-made automated platform for high throughput processing (HTP) of field-trapped sandflies. The aim is to present an integrated approach to discover new/existing viruses vectored by sandflies, to characterize these viruses by complete genome sequencing, and to study their potential public health impact for humans and non human vertebrates. Sandflies were collected in France, Tunisia, Algeria, Turkey and Iran. They were processed on a custom-made automated HTP platform. RT-PCR assays targeting conserved regions within the genus Phlebovirus were performed both directly on sandfly extracted RNA and on supernatant of Vero cells, and further sequenced. Sandflies and cell culture that were PCR-positive, as well as supernatant of flasks with cytopathic effect were sequenced using a de novo protocol onto a Ion Torrent PGM sequencer. A total of 29,664 sandflies were collected and processed onto the platform. Origin and number of specimens are as follows: France (n = 3,087), Tunisia (n = 8,206), Algeria (n = 1,283), Turkey (n = 12,000), Iran (n = 5,089). We isolated 9 virus strains in France (2 Toscana, 7 Massilia), 10 strains in Tunisia (2 Toscana, 5 Punique, 3 new viruses still unnamed), 1 strain in Algeria (1 Toscana), 2 strains in Turkey (new phlebovirus, provisionally named Damyeri), 1 strain in Iran (new phlebovirus, provisionally named Dashli). Analysis using complete gene sequences allowed to produce full protein phylograms for isolated virus strains. Seroprevalence studies were performed to define whether these viruses infect humans and non human vertebrates in regions where sandflies were trapped or not. Seroprevalence results will be presented for France and Tunisia. A total of 23 strains were isolated, 6 of which correspond to new sandfly-borne phleboviruses that have been completely sequenced through NGS. Screening field-trapped sandflies is suitable to inventory recognized and new viruses. Combining cell culture for isolation and NGS for sequencing is cost-effective and adapted to process high quantities of insects. Serological tools are a rapid and easy method to address potential public health impact which must be confirmed by molecular diagnostic derived from complete sequence data obtained through NGS.

Toscana virus (TOSV) was detected for the first time from Phlebotomus perniciosus sandflies in Corsica, a French Mediterranean island. Genetic analysis showed that Corsican TOSV belongs to lineage A, together with Italian, Tunisian, Turkish and other French strains. The demonstration of TOSV in Corsica indicates that autochthonous and tourist populations are at risk of infection. Hence, physicians must consider TOSV as a possible cause of aseptic meningitis and unidentified febrile illness during the warm season.

Rapid documentation of respiratory specimens can have an impact on the management of patients and their relatives in terms of preventive and curative measures. We compared the results of the Xpert® Flu assay (Cepheid) with three real-time RT-PCR assays using 127 nasopharyngeal samples, of which 75 were positive for influenza A (with 52 identified as A/HINI-2009) and 52 were positive for influenza B. The Xpert® Flu assay presented a quasi-absence of non-interpretable tests, and showed sensitivity and specificity of 100% and 100% for Flu A, 98.4% and 100% for A/HINI-2009, and 80.7% and 100% for Flu B.

We describe a fatal case of Reye's syndrome in a 12-year-old male patient during an influenza A (H3N2) infection for which he received salicylates. In the current situation of the novel A/H1N1 virus pandemic, we believe that it is of high importance to emphasize the risks associated with salicylate intake to avoid the reappearance of Reye's syndrome.

R. N. Charrel, I. Leparc-Goffart, P. Gallian and X. de Lamballerie 1) Aix Marseille Universit e, IRD French Institute of Research for Development, EHESP French School of Public Health, EPV UMR_D 190 ‘Emergence des Pathologies Virales’, IHU M editerran ee Infection, APHM Public Hospitals of Marseille, Marseille, 2) National Reference Laboratory for Arboviruses, Institut de Recherche Biom edicale des Arm ees, Marseille and 3) Etablissement Franc ais du Sang Alpes-M editerran ee, Marseille, France E-mail: remi.charrel@univ-amu.fr Article published online: 28 May 2014

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.

Management of the first wave of the H1N1 influenza pandemic has presented major logistic and infrastructural challenges for public health authorities worldwide. Mangement in the Northern hemisphere has been in part guided by information provided by countries in the Southern hemisphere, which were the first to experience a winter outbreak of the new influenza H1N1 variant virus. Following reports from Australia and New Zealand, a recent symposium organized in Paris by the French health authorities (November 19, 2009; http://www.invs.sante.fr/behweb/2009/04/index.htm) gathered all the parties involved in management of the epidemics in French overseas territories located in the Southern hemisphere, i.e. New Caledonia, French Polynesia and Wallis and Futuna islands (Pacific Ocean) and Reunion and Mayotte islands (Indian Ocean). An assessment was produced by the Institut de Veille Sanitaire (InVS, the French institute in charge of public health surveillance). The experts concluded that the impact of the influenza outbreak had been more severe than previous seasonal epidemics, with estimated attack rates varying between 8% and 18%. All territories separated infected and non-infected patient admissions in hospitals and outpatient departments. The duration of the epidemic wave was short (8–12 weeks) and the peak incidence was generally observed at the 4th or 5th week. In Mayotte island, virus circulation was longer, a possible consequence of the typical tropical climate. Globally, all experts agreed that the common clinical presentation was mild but the frequency of severe forms or Acute Respiratory Distress Syndrome (ARDS) was unusually high, with a significant proportion (10–20%) of patients admitted to ICUs because of bacterial superinfection (mainly pneumococcus or Haemophilus influenzae). While the health systems were overwhelmed in none of the French overseas territories, it was considered likely that in new Caledonia and French Polynesia, one additional week of case reports at the peak level would have led to worrying saturation of ICUs. Thus the relatively limited availability of Extracorporeal Membrane Oxygenation (ECMO) machines for resuscitation by oxygenation and trained staff was also a cause for concern. As far as virological diagnosis was concerned, all territories reported saturation of laboratory capacity and, for some (e.g. Wallis and Futuna), prolonged delays in obtaining results, due to the absence of local diagnostic facilities. In addition, diagnosis of influenza often competed with other microbiological diagnoses which were also delayed for similar reasons. In this context, the availability of diagnostic tests and in particular the possible use and reliability of rapid influenza diagnostic tests (RIDTs) were important limiting factors. In the current issue, we present reports from the Pacific zone (New Caledonia, French Polynesia and Wallis and Futuna islands; see the paper by La Ruche et al.) and from the Indian Ocean zone (Reunion and Mayotte islands; see the paper by D’Ortenzio et al.) that detail the epidemiological analysis of the first pandemic wave. It should be noted that seroprevalence studies are still required to evaluate precisely the actual infection rate in these territories, an important prerequisite for devising a robust strategy for the preparation and management of subsequent influenza epidemic waves. Preliminary data suggest a high proportion of asymptomatic cases, but a precise assessment is still awaited. The current issue also includes new technical diagnostic developments (see the paper by Bouscambert Duchamp et al.) and data suggesting that the use of some (but not all) commercially available RIDTs could contribute significantly to the rapid management of suspected cases (see the paper by Nougairede et al.). In addition (see the papers by Nougairede et al. and by Casalegno et al.), interim reports on the A/ H1N1 influenza virus pandemic in mainland France are presented, including information regarding possible viral interference between respiratory viruses that could have affected the spread of A/H1N1 influenza virus and delayed the influenza pandemic in France.