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a new permanent cell line derived from the bank vole (myodes glareolus) as cell culture model for zoonotic viruses.abstract:201121729307
Usutu virus: potential risk of human disease in Europe.Usutu virus (USUV) is an African mosquito-borne flavivirus, member of the Japanese encephalitis antigenic group. This avian virus is transmitted by arthropod vectors (mainly mosquitoes of the Culex pipiens complex). It is well known that free-living birds, including migratory species, have the potential to disperse certain pathogenic microorganisms. Usutu virus has recently been introduced to Europe and is spreading through Austria, Hungary, Italy, Spain and Switzerland, causing disease in birds ...201121871214
phylogenetic analysis of west nile virus isolates, italy, 2008-2009.to determine the lineage of west nile virus that caused outbreaks in italy in 2008 and 2009, several west nile virus strains were isolated from human specimens and sequenced. on the basis of phylogenetic analyses, the strains isolated constitute a distinct group within the western mediterranean cluster.201121529408
application of in-situ hybridization for the detection and identification of avian malaria parasites in paraffin wax-embedded tissues from captive penguins.in captive penguins, avian malaria due to plasmodium parasites is a well-recognized disease problem as these protozoa may cause severe losses among valuable collections of zoo birds. in blood films from naturally infected birds, identification and differentiation of malaria parasites based on morphological criteria are difficult because parasitaemia is frequently light and blood stages, which are necessary for identification of parasites, are often absent. post-mortem diagnosis by histological e ...201121711191
bagaza virus in partridges and pheasants, spain, 2010.in september 2010, an unusually high number of wild birds (partridges and pheasants) died in cádiz in southwestern spain. reverse transcription pcr and virus isolation detected flavivirus infections. complete nucleotide sequence analysis identified bagaza virus, a flavivirus with a known distribution that includes sub-saharan africa and india, as the causative agent.201121801633
detection of usutu virus within a west nile virus surveillance program in northern italy.abstract usutu virus (usuv) is a mosquito-borne flavivirus belonging to the japanese encephalitis serocomplex, recently related to neurological disease in immunosuppressed patients. in the same area of northern italy where usuv human cases occurred in 2009, a regional west nile virus (wnv) surveillance program based on mosquito monitoring and wild birds screening has been implemented since 2008. mosquito pools and wild birds were tested using three different polymerase chain reactions (flaviviru ...201120849275
west nile virus circulation in veneto region in 2008-2009.summarywest nile virus (wnv) was detected in italy, in late summer 2008 in horses and birds in the po valley. as a consequence, an intense wnv surveillance was implemented in that area involving emilia-romagna, veneto and lombardy. this paper presents the results of the september 2008-november 2009 surveillance on equines, mosquitoes, wild birds, dogs and cattle in veneto. wnv was detected in equines and dogs, and, to a lesser extent in cattle and wild birds. simultaneous circulation of usutu vi ...201120670469
west nile virus population genetics and evolution.west nile virus (wnv) (flaviviridae: flavivirus) is transmitted from mosquitoes to birds, but can cause fatal encephalitis in infected humans. since its introduction into north america in new york in 1999, it has spread throughout the western hemisphere. multiple outbreaks have also occurred in europe over the last 20 years. this review highlights recent efforts to understand how host pressures impact viral population genetics, genotypic and phenotypic changes which have occurred in the wnv geno ...201122226703
west nile virus replication requires fatty acid synthesis but is independent on phosphatidylinositol-4-phosphate lipids.west nile virus (wnv) is a neurovirulent mosquito-borne flavivirus, which main natural hosts are birds but it also infects equines and humans, among other mammals. as in the case of other plus-stranded rna viruses, wnv replication is associated to intracellular membrane rearrangements. based on results obtained with a variety of viruses, different cellular processes have been shown to play important roles on these membrane rearrangements for efficient viral replication. as these processes are re ...201121949814
structure based drug designing of a novel antiflaviviral inhibitor for nonstructural 3 protein.literature shows that flaviviruses cause a variety of diseases, including fevers, encephalitis, and hemorrhagic fevers. ns3 is a multifunctional protein with an nterminal protease domain (ns3pro) that is responsible for proteolytic processing of the viral polyprotein, and a c-terminal region that contains an rna triphosphatase, rna helicase and rna-stimulated ntpase domain that are essential for rna replication. therefore, ns3 protein is the preferential choice for inhibition to stop the proteol ...201121544165
rapid molecular detection methods for arboviruses of livestock of importance to northern europe.arthropod-borne viruses (arboviruses) have been responsible for some of the most explosive epidemics of emerging infectious diseases over the past decade. their impact on both human and livestock populations has been dramatic. the early detection either through surveillance or diagnosis of virus will be a critical feature in responding and resolving the emergence of such epidemics in the future. although some of the most important emerging arboviruses are human pathogens, this paper aims to high ...201222219660
assessing the risks of west nile virus-infected mosquitoes from transatlantic aircraft: implications for disease emergence in the united kingdom.abstract the number of west nile virus (wnv)-infected mosquitoes aboard aircraft from the united states that arrive in the united kingdom each summer was determined using a quantitative risk assessment. in the worst-case scenario, when wnv levels in mosquitoes are high (at epidemic levels) the probability of at least one wnv-infected mosquito being introduced into the united kingdom was predicted to be 0.99. during these periods, a mean of 5.2 infected mosquitoes were estimated to be aboard fl ...201222217181
feeding patterns of potential west nile virus vectors in south-west spain.mosquito feeding behaviour determines the degree of vector-host contact and may have a serious impact on the risk of west nile virus (wnv) epidemics. feeding behaviour also interacts with other biotic and abiotic factors that affect virus amplification and transmission.201222745781
animal viral diseases and global change: bluetongue and west nile fever as paradigms.environmental changes have an undoubted influence on the appearance, distribution, and evolution of infectious diseases, and notably on those transmitted by vectors. global change refers to environmental changes arising from human activities affecting the fundamental mechanisms operating in the biosphere. this paper discusses the changes observed in recent times with regard to some important arboviral (arthropod-borne viral) diseases of animals, and the role global change could have played in th ...201222707955
epizootic emergence of usutu virus in wild and captive birds in germany.this study aimed to identify the causative agent of mass mortality in wild and captive birds in southwest germany and to gather insights into the phylogenetic relationship and spatial distribution of the pathogen. since june 2011, 223 dead birds were collected and tested for the presence of viral pathogens. usutu virus (usuv) rna was detected by real-time rt-pcr in 86 birds representing 6 species. the virus was isolated in cell culture from the heart of 18 blackbirds (turdus merula). usuv-specif ...201222389712
approaches for the development of rapid serological assays for surveillance and diagnosis of infections caused by zoonotic flaviviruses of the japanese encephalitis virus serocomplex.flaviviruses are responsible for a number of important mosquito-borne diseases of man and animals globally. the short vireamic period in infected hosts means that serological assays are often the diagnostic method of choice. this paper will focus on the traditional methods to diagnose flaviviral infections as well as describing the modern rapid platforms and approaches for diagnostic antigen preparation.201222570528
west nile virus: characterization and diagnostic applications of monoclonal antibodies.diagnosis of west nile virus (wnv) infections is often difficult due to the extensive antigenic cross-reactivity among flaviviruses, especially in geographic regions where two or more of these viruses are present causing sequential infections. the purpose of this study was to characterize a panel of monoclonal antibodies (mabs) produced against wnv to verify their applicability in wnv diagnosis and in mapping epitope targets of neutralizing mabs.201222500562
development of monoclonal antibodies to west nile virus and their application in immunohistochemistry.west nile virus (wnv) is endemic throughout africa, eurasia, america, and australia and has important implications for avian, horse, and human health. in these regions, dead birds are monitored for the presence of wnv through immunohistochemistry (ihc) and pcr. however, a number of the tools for ihc are inadequate owing to their cross-reactivity to other japanese encephalitis serogroup viruses. here we have established eight monoclonal antibodies (mabs) to wnv. four of them bound to the envelope ...201222993408
[when some flaviviruses are throwing our certainties].during the past two decades, a number of mosquito-borne flaviviruses, mainly of african origin, have invaded new geographical areas where they have never been active. this was the case for the japanese encephalitis virus which reached the northeastern part of australia (1995) and, above all, for the west nile virus which, since 1999, entirely colonized the american continent. then, the usutu virus invaded a large part of the western europe (2001) while the zika virus caused a large epidemic in a ...201222923343
real-time fluorogenic reverse transcription polymerase chain reaction assay for the specific detection of bagaza virus.in september 2010, an outbreak of disease in 2 wild bird species (red-legged partridge, alectoris rufa; ring-necked pheasant, phasianus colchicus) occurred in southern spain. bagaza virus (bagv) was identified as the etiological agent of the outbreak. bagv had only been reported before in western africa (central african republic, senegal) and in india. the first occurrence of bagv in spain stimulated a demand for rapid, reliable, and efficacious diagnostic methods to facilitate the surveillance ...201222807508
anopheles plumbeus (diptera: culicidae) in europe: a mere nuisance mosquito or potential malaria vector?anopheles plumbeus has been recognized as a minor vector for human malaria in europe since the beginning of the 20th century. in recent years this tree hole breeding mosquito species appears to have exploited novel breeding sites, including large and organically rich man-made containers, with consequently larger mosquito populations in close vicinity to humans. this lead to investigate whether current populations of an. plumbeus would be able to efficiently transmit plasmodium falciparum, the pa ...201223181931
how often do they meet? genetic similarity between european populations of a potential disease vector culex pipiens.species in the culex pipiens complex are common almost all over the world and represent important vectors for many serious zoonotic diseases. even if, at the moment, many of the pathogens potentially transmitted by cx. pipiens are not a problem in northern europe, they may, with increasing temperatures and changing ecosystems caused by climate change, move northward in the future. therefore, the question whether or not the cx. pipiens populations in northern europe will be competent vectors for ...201222957132
japanese encephalitis virus rna detected in culex pipiens mosquitoes in italy.mosquitoes collected in northern italy were screened for flavivirus rna. positive amplicons were sequenced and found most similar to insect flavivirus (isf), usutu virus (usuv) and surprisingly also to japanese encephalitis virus (jev). the sequence (167 bp), obtained from one pool of culex pipiens, was found identical to jev strains from bats in china. unfortunately additional sequence data or virus isolations were not obtained in this study. confirmation of potential introduction of jev to ita ...201222835438
detection of usutu virus infection in a healthy blood donor from south-west germany, 2012.from september 2011 until november 2012, 31 serum samples from german patients with clinically suspected acute usutu virus (usuv) infections were tested for usuv-specific antibodies. all samples tested negative. in addition, 4,200 serum samples from healthy blood donors from south-west germany were collected in january 2012 and also analysed for the presence of specific antibodies. one sample tested positive for usuv-igg and -igm. thus, the seroprevalence of usuv antibodies in healthy blood dono ...201223241231
genetic diversity of usutu virus.usutu virus is a mosquito-borne virus first isolated from culex naevei in south africa in 1959. the first emergence of usutu virus outside africa was recorded in austria. here, a phylogenetic analysis targeting the e5 and ns5 genes was carried out on the viral strains circulating in europe. the ns5 gene tree showed two main clades, one of which included the italian sequences. in the e gene tree all sequences grouped into the same main clade, with sequences from austria divided into two separate ...201222707129
blood meal analysis, flavivirus screening, and influence of meteorological variables on the dynamics of potential mosquito vectors of west nile virus in northern italy.an extended area of northern italy has experienced several west nile virus (wnv) outbreaks and the emergence of usutu virus (usuv) during previous years. our aim was to study some of the factors that could explain disease patterns in the trentino region, where circulation was detected in human sera and sentinel chickens, but no human or equine cases were reported. we collected culex species (diptera: culicidae) in peridomestic environments. the collected specimens were analyzed for feeding behav ...201222548533
a diagnostic algorithm to serologically differentiate west nile virus from japanese encephalitis virus infections and its validation in field surveillance of poultry and horses.abstract the detection of west nile virus (wnv) in areas endemic for japanese encephalitis virus (jev) is complicated by the extensive serological cross-reactivity between the two viruses. a testing algorithm was developed and employed for the detection of anti-wnv antibody in areas endemic for jev. using this differentiation algorithm, a serological survey of poultry (2004 through 2009) and horses (2007 through 2009) was performed. among 2681 poultry sera, 125 samples were interpreted as bein ...201222217162
detection of usutu-virus-specific igg in blood donors from northern italy.abstract we developed a novel enzyme-linked immunosorbent assay to detect the specific igg response to usutu virus (usuv) in humans, by evaluating 359 blood donors who were living in northeastern italy. our results demonstrate the presence of an anti-usuv response in 4 subjects with no history of other flavivirus infection.201222217176
evolution of mosquito-based arbovirus surveillance systems in australia.control of arboviral disease is dependent on the sensitive and timely detection of elevated virus activity or the identification of emergent or exotic viruses. the emergence of japanese encephalitis virus (jev) in northern australia revealed numerous problems with performing arbovirus surveillance in remote locations. a sentinel pig programme detected jev activity, although there were a number of financial, logistical, diagnostic and ethical limitations. a system was developed which detected vir ...201222505808
mosquito, bird and human surveillance of west nile and usutu viruses in emilia-romagna region (italy) in 2010.in 2008, after the first west nile virus (wnv) detection in the emilia-romagna region, a surveillance system, including mosquito- and bird-based surveillance, was established to evaluate the virus presence. surveillance was improved in following years by extending the monitoring to larger areas and increasing the numbers of mosquitoes and birds tested.201222666446
evidence of mosquito-transmitted flavivirus circulation in piedmont, north-western italy.flavivirus is a highly heterogeneous viral genus that includes important human pathogens and several viral strains with unknown zoonotic potential. mosquito-borne flaviviruses have been isolated and characterized in northern italy: west nile virus and usutu virus were detected in mosquitoes and in different host species and recent studies provided evidence about the circulation of "insect flavivirus" strains.201222616748
the contribution of rodent models to the pathological assessment of flaviviral infections of the central nervous system.members of the genus flavivirus are responsible for a spectrum of important neurological syndromes in humans and animals. rodent models have been used extensively to model flavivirus neurological disease, to discover host-pathogen interactions that influence disease outcome, and as surrogates to determine the efficacy and safety of vaccines and therapeutics. in this review, we discuss the current understanding of flavivirus neuroinvasive disease and outline the host, viral and experimental facto ...201222592957
hybridization and population structure of the culex pipiens complex in the islands of macaronesia.the culex pipiens complex includes two widespread mosquito vector species, cx. pipiens and cx. quinquefasciatus. the distribution of these species varies in latitude, with the former being present in temperate regions and the latter in tropical and subtropical regions. however, their distribution range overlaps in certain areas and interspecific hybridization has been documented. genetic introgression between these species may have epidemiological repercussions for west nile virus (wnv) transmis ...201222957190
european surveillance for west nile virus in mosquito populations.a wide range of arthropod-borne viruses threaten both human and animal health either through their presence in europe or through risk of introduction. prominent among these is west nile virus (wnv), primarily an avian virus, which has caused multiple outbreaks associated with human and equine mortality. endemic outbreaks of west nile fever have been reported in italy, greece, france, romania, hungary, russia and spain, with further spread expected. most outbreaks in western europe have been due ...201324157510
zoonoses in pet birds: review and perspectives.pet birds are a not-so-well known veterinarian's clientship fraction. bought individually or in couples, as families often do (which is a lucrative business for pet shops or local breeders) or traded (sometimes illegally) for their very high genetic or exotic value, these birds, commonly canaries, parakeets or parrots, are regularly sold at high prices. these animals, however, are potential carriers and/or transmitters of zoonotic diseases. some of them could have an important impact on human he ...201323687940
could drought conditions trigger schmallenberg virus and other arboviruses circulation?in 2011, a new orthobunyavirus, named the schmallenberg virus (sbv), was discovered in europe. like the related shamonda virus, sbv is an arbovirus (arthropod-borne virus). after its discovery, the virus was detected in a wide area in north-western europe, an unexpected finding in a territory where climatic conditions would not seem ideal for arbovirus transmission. this sudden expansion suggests the effect of 2011 drought as a key factor that may have triggered sbv circulation. the possible inf ...201323409725
intertwined arbovirus transmission activity: reassessing the transmission cycle paradigm.arboviruses are emerging/reemerging infectious agents worldwide. the factors within this scenario include vector and host population fluctuations, climatic changes, anthropogenic activities that disturb ecosystems, an increase in international flights, human mobility, and genetic mutations that allow spill-over phenomenon. arboviruses are maintained by biologic transmission among vectors and hosts. sometimes this biological transmission is specific and includes one vector and host species such a ...201323335900
novel cis-acting element within the capsid-coding region enhances flavivirus viral-rna replication by regulating genome cyclization.cis-acting elements in the viral genome rna (vrna) are essential for the translation, replication, and/or encapsidation of rna viruses. in this study, a novel conserved cis-acting element was identified in the capsid-coding region of mosquito-borne flavivirus. the downstream of 5' cyclization sequence (5'cs) pseudoknot (dcs-pk) element has a three-stem pseudoknot structure, as demonstrated by structure prediction and biochemical analysis. using dengue virus as a model, we show that dcs-pk enhanc ...201323576500
zoonotic encephalitides caused by arboviruses: transmission and epidemiology of alphaviruses and flaviviruses.in this review, we mainly focus on zoonotic encephalitides caused by arthropod-borne viruses (arboviruses) of the families flaviviridae (genus flavivirus) and togaviridae (genus alphavirus) that are important in both humans and domestic animals. specifically, we will focus on alphaviruses (eastern equine encephalitis virus, western equine encephalitis virus, venezuelan equine encephalitis virus) and flaviviruses (japanese encephalitis virus and west nile virus). most of these viruses were origin ...201324427764
usutu virus persistence and west nile virus inactivity in the emilia-romagna region (italy) in 2011.the circulation of west nile virus and usutu virus was detected in the emilia-romagna region in 2008 and 2009. to evaluate the extent of circulation of both viruses, environmental surveillance, based on bird and mosquito testing, was conducted in 2008 and gradually improved over the years.201323667694
identification of hotspots in the european union for the introduction of four zoonotic arboviroses by live animal trade.live animal trade is considered a major mode of introduction of viruses from enzootic foci into disease-free areas. due to societal and behavioural changes, some wild animal species may nowadays be considered as pet species. the species diversity of animals involved in international trade is thus increasing. this could benefit pathogens that have a broad host range such as arboviruses. the objective of this study was to analyze the risk posed by live animal imports for the introduction, in the e ...201323894573
vector-virus interactions and transmission dynamics of west nile virus.west nile virus (wnv; flavivirus; flaviviridae) is the cause of the most widespread arthropod-borne viral disease in the world and the largest outbreak of neuroinvasive disease ever observed. mosquito-borne outbreaks are influenced by intrinsic (e.g., vector and viral genetics, vector and host competence, vector life-history traits) and extrinsic (e.g., temperature, rainfall, human land use) factors that affect virus activity and mosquito biology in complex ways. the concept of vectorial capacit ...201324351794
use of competition elisa for monitoring of west nile virus infections in horses in germany.west nile virus (wnv) is a mosquito-borne viral pathogen of global importance and is considered to be the most widespread flavivirus in the world. horses, as dead-end hosts, can be infected by bridge mosquito vectors and undergo either subclinical infections or develop severe neurological diseases. the aim of this study was to detect wnv specific antibodies in horses in germany as an indicator for an endemic circulation of wnv. sera from more than 5,000 horses (primarily fallen stock animals) we ...201323887620
a single amino acid substitution in the core protein of west nile virus increases resistance to acidotropic compounds.west nile virus (wnv) is a worldwide distributed mosquito-borne flavivirus that naturally cycles between birds and mosquitoes, although it can infect multiple vertebrate hosts including horses and humans. this virus is responsible for recurrent epidemics of febrile illness and encephalitis, and has recently become a global concern. wnv requires to transit through intracellular acidic compartments at two different steps to complete its infectious cycle. these include fusion between the viral enve ...201323874963
ilheus virus isolation in the pantanal, west-central brazil.the wetlands of the brazilian pantanal host large concentrations of diverse wildlife species and hematophagous arthropods, conditions that favor the circulation of zoonotic arboviruses. a recent study from the nhecolândia sub-region of pantanal reported serological evidence of various flaviviruses, including west nile virus and ilheus virus (ilhv). according to the age of seropositive horses, at least three flaviviruses, including ilhv, circulated in the brazilian pantanal between 2005 and 2009. ...201323875051
west nile virus. 201324179475
mosquito-borne arbovirus surveillance at selected sites in diverse ecological zones of kenya; 2007 - 2012.increased frequency of arbovirus outbreaks in east africa necessitated the determination of distribution of risk by entomologic arbovirus surveillance. a systematic vector surveillance programme spanning 5 years and covering 11 sites representing seven of the eight provinces in kenya and located in diverse ecological zones was carried out.201323663381
a genome-wide analysis of rna pseudoknots that stimulate efficient -1 ribosomal frameshifting or readthrough in animal viruses.programmed -1 ribosomal frameshifting (prf) and stop codon readthrough are two translational recoding mechanisms utilized by some rna viruses to express their structural and enzymatic proteins at a defined ratio. efficient recoding usually requires an rna pseudoknot located several nucleotides downstream from the recoding site. to assess the strategic importance of the recoding pseudoknots, we have carried out a large scale genome-wide analysis in which we used an in-house developed program to d ...201324298557
potential changes in disease patterns and pharmaceutical use in response to climate change.as climate change alters environmental conditions, the incidence and global patterns of human diseases are changing. these modifications to disease profiles and the effects upon human pharmaceutical usage are discussed. climate-related environmental changes are associated with a rise in the incidence of chronic diseases already prevalent in the northern hemisphere, for example, cardiovascular disease and mental illness, leading to greater use of associated heavily used western medications. suffe ...201323909463
ontology for vector surveillance and management.ontologies, which are made up by standardized and defined controlled vocabulary terms and their interrelationships, are comprehensive and readily searchable repositories for knowledge in a given domain. the open biomedical ontologies (obo) foundry was initiated in 2001 with the aims of becoming an "umbrella" for life-science ontologies and promoting the use of ontology development best practices. a software application (obo-edit; *.obo file format) was developed to facilitate ontology developmen ...201323427646
duck tembusu virus exhibits neurovirulence in balb/c mice.duck tembusu virus is a member of the ntaya group in the genus flavivirus. the virus has been responsible for severe duck egg-drop syndrome in china since 2010. its emergence and rapid spread have caused great economic loss for the poultry industry. the epidemiology of the virus infection and the potential threat to public health is of great concern because of the infective and zoonotic nature of flaviviruses.201323941427
targeted surveillance for usutu virus in british birds (2005-2011). 201323193038
detection of specific antibodies against west nile and usutu viruses in healthy blood donors in northern italy, 2010-2011.neutralizing antibodies against west nile (wnv) and usutu (usuv) viruses were measured in 6000 samples collected, between 1 september 2010 and 30 june 2011, from blood donors living in different districts of emilia-romagna, northeastern italy. on the basis of the microneutralization assay (mnta), 47 (0.78%) subjects were positive for wnv and 14 (0.23%) for usuv. these results were compared with those obtained 2 years ago and suggest an increased circulation of usuv among humans in emilia-romagna ...201323663225
monitoring west nile virus (wnv) infection in wild birds in serbia during 2012: first isolation and characterisation of wnv strains from serbia.west nile virus (wnv), a neurovirulent mosquito-transmissible zoonotic virus, has caused recent outbreaks in europe, including serbia from august until october 2012. although humans can be infected, birds are the main natural wnv reservoir. to assess wnv circulation in northern serbia, 133 wild birds were investigated. these comprised resident and migratory birds, collected between january and september 2012 in the vojvodina province. the birds belonged to 45 species within 27 families. blood se ...201324176657
retrospective identification of human cases of west nile virus infection in austria (2009 to 2010) by serological differentiation from usutu and other flavivirus infections.there is increasing evidence for the spread of west nile virus (wnv) in southern, eastern and central europe. in parallel, another flavivirus, the antigenically closely related usutu virus, was introduced from africa and first detected in austria (2001), followed by spain (2003), hungary (2005), italy (2006), switzerland (2006) and germany (2007). in austria, human wnv infections have not previously been documented, although the virus was isolated from birds and detected in mosquitoes in 2008 an ...201324176619
a novel quantitative multiplex real-time rt-pcr for the simultaneous detection and differentiation of west nile virus lineages 1 and 2, and of usutu virus.an increase in activity of two mosquito-borne flaviviruses, west nile virus (wnv) and usutu virus (usuv), has been reported in europe in recent years. the current epidemiological situation calls for rt-pcr methods that are able to detect not only the widespread lineage 1 (l1) wnv, but also lineage 2 (l2) wnv. in addition, the presence in europe of the closely related usuv requires methods that can identify these three flaviviruses and permit an efficient and accurate differential diagnosis. here ...201323499258
usutu virus growth in human cell lines: induction of and sensitivity to type i and iii interferons.the mechanisms of usutu virus (usuv) pathogenesis are largely unknown. the aim of this study was to evaluate the sensitivity of usuv to interferon (ifn) and the capacity of usuv to stimulate ifn production. initial experiments were conducted to characterize the susceptibility of human cell lines to usuv infection and to evaluate the single-growth cycle replication curve of usuv. results indicate that usuv is able to infect a variety of human cell lines, completing the replication cycle in hep-2 ...201323255619
bluetongue disease risk assessment based on observed and projected culicoides obsoletus spp. vector densities.bluetongue is an arboviral disease of ruminants causing significant economic losses. our risk assessment is based on the epidemiological key parameter, the basic reproduction number. it is defined as the number of secondary cases caused by one primary case in a fully susceptible host population, in which values greater than one indicate the possibility, i.e., the risk, for a major disease outbreak. in the course of the bluetongue virus serotype 8 (btv-8) outbreak in europe in 2006 we developed s ...201323560090
effect of blood meal digestion and dna extraction protocol on the success of blood meal source determination in the malaria vector anopheles atroparvus.host identification is an essential step in studies on the transmission dynamics of vector-borne disease. nowadays, molecular tools allow the identification of vertebrate hosts to the species level. however, the proportion of successful identifications is variable and may be affected by the quality of the samples and the laboratory protocols. here, the effect of two of these factors, namely the digestion status of mosquito blood meal and the dna extraction procedure, on the success of host ident ...201323517864
demonstration of usutu virus antibodies in horses, croatia.we report the first serological evidence of usutu virus (usuv) infection in horses in croatia. during 2011, 1380 horse serum samples from healthy animals were collected from six northern croatian counties. all samples were first screened for west nile virus (wnv) immunoglobulin g (igg) antibodies using an enzyme-linked immunosorbent assay (elisa). sixty-nine wnv elisa-reactive samples were further tested for wnv antibodies by a virus neutralization assay (vn assay) and plaque reduction neutraliz ...201323808977
lack of identification of flaviviruses in oral and cloacal swabs from long- and short-distance migratory birds in trentino-alto adige (north-eastern italy).west nile virus (wnv) and usutu virus (usuv), both belonging to the genus flavivirus, are emerging in italy as important human and animal pathogens. migratory birds are involved in the spread of flaviviruses over long distances, particularly from africa to europe. once introduced, these viruses can be further be dispersed by short-distance migratory and resident bird species. thus far, there is still a considerable knowledge gap on the role played by different bird species in the ecology and tra ...201324119320
pathogen-host associations and predicted range shifts of human monkeypox in response to climate change in central africa.climate change is predicted to result in changes in the geographic ranges and local prevalence of infectious diseases, either through direct effects on the pathogen, or indirectly through range shifts in vector and reservoir species. to better understand the occurrence of monkeypox virus (mpxv), an emerging orthopoxvirus in humans, under contemporary and future climate conditions, we used ecological niche modeling techniques in conjunction with climate and remote-sensing variables. we first crea ...201323935820
second international diagnostic accuracy study for the serological detection of west nile virus infection.in recent decades, sporadic cases and outbreaks in humans of west nile virus (wnv) infection have increased. serological diagnosis of wnv infection can be performed by enzyme-linked immunosorbent assay (elisa), immunofluorescence assay (ifa) neutralization test (nt) and by hemagglutination-inhibition assay. the aim of this study is to collect updated information regarding the performance accuracy of wnv serological diagnostics.201323638205
usutu virus. 201324659950
phylogeography of japanese encephalitis virus: genotype is associated with climate.the circulation of vector-borne zoonotic viruses is largely determined by the overlap in the geographical distributions of virus-competent vectors and reservoir hosts. what is less clear are the factors influencing the distribution of virus-specific lineages. japanese encephalitis virus (jev) is the most important etiologic agent of epidemic encephalitis worldwide, and is primarily maintained between vertebrate reservoir hosts (avian and swine) and culicine mosquitoes. there are five genotypes o ...201324009790
low usutu virus seroprevalence in four zoological gardens in central europe.usutu virus (usuv), a mosquito-borne flavivirus of the japanese encephalitis virus antigenic group, caused bird die-offs in austria, hungary and switzerland between 2001 and 2009. while the zoological gardens of vienna and zurich recorded usuv-associated mortality in different species of birds during this period, incidences in budapest were limited to areas outside the zoo, and in the greater basel area avian mortality due to usuv infection was not observed at all. the objectives of this investi ...201323919825
west nile virus equine serosurvey in the czech and slovak republics.a serological survey for west nile virus (wnv) infection involved 395 horses from 43 administrative districts of the czech republic (163 animals) and 29 districts of slovakia (232 animals), sampled between 2008 and 2011. using a plaque-reduction neutralization microtest, antibodies to wnv were not detected in any horse from the czech republic, whereas 19 nonvaccinated horses from slovakia had specific antibodies to wnv (no cross-reactions were observed with tick-borne encephalitis and usutu flav ...201323919605
southernmost asia is the source of japanese encephalitis virus (genotype 1) diversity from which the viruses disperse and evolve throughout asia.although a previous study predicted that japanese encephalitis virus (jev) originated in the malaysia/indonesia region, the virus is known to circulate mainly on the asian continent. however, there are no reported systematic studies that adequately define how jev then dispersed throughout asia.201324069502
infection with usutu virus induces an autophagic response in mammalian cells.usutu virus (usuv) is an african mosquito-borne flavivirus closely related to west nile virus and japanese encephalitis virus, which host range includes mainly mosquitoes and birds, although infections in humans have been also documented, thus warning about usuv as a potential health threat. circulation of usuv in africa was documented more than 50 years ago, but it was not until the last decade that it emerged in europe causing episodes of avian mortality and some human severe cases. since auto ...201324205422
population seroprevalence study after a west nile virus lineage 2 epidemic, greece, 2010.during summer 2010, 262 human cases including 35 deaths from west nile virus (wnv) infection were reported from central macedonia, greece. evidence from mosquitoes, birds and blood donors demonstrated that the epidemic was caused by wnv lineage 2, which until recently was considered of low virulence. we conducted a household seroprevalence study to estimate the spread of infection in the population during the epidemic, ascertain the relationship of infection to clinical disease, and identify ris ...201324260390
first outbreak of west nile virus neuroinvasive disease in humans, croatia, 2012.between september 6 and 21, 2012, seven human cases of west nile virus (wnv) neuroinvasive infection were laboratory confirmed in croatia. the median patient age was 62 years (range 48-77). five patients presented with meningoencephalitis and two patients with meningoencephalitis followed by acute flaccid paralysis. four of them had an underlying disease (hypertension). using enzyme-linked immunosorbent assay (elisa), wnv-specific immunoglobulin m (igm) and igg antibodies of low avidity were det ...201324283515
climate change and public health policy: translating the science.public health authorities are required to prepare for future threats and need predictions of the likely impact of climate change on public health risks. they may get overwhelmed by the volume of heterogeneous information in scientific articles and risk relying purely on the public opinion articles which focus mainly on global warming trends, and leave out many other relevant factors. in the current paper, we discuss various scientific approaches investigating climate change and its possible impa ...201324452252
flaviviruses in europe: complex circulation patterns and their consequences for the diagnosis and control of west nile disease.in europe, many flaviviruses are endemic (west nile, usutu, tick-borne encephalitis viruses) or occasionally imported (dengue, yellow fever viruses). due to the temporal and geographical co-circulation of flaviviruses in europe, flavivirus differentiation by diagnostic tests is crucial in the adaptation of surveillance and control efforts. serological diagnosis of flavivirus infections is complicated by the antigenic similarities among the flavivirus genus. indeed, most flavivirus antibodies are ...201324225644
development of one-step quantitative reverse transcription pcr for the rapid detection of flaviviruses.the genus flavivirus includes several pathogenic agents that cause severe illness in humans. re-emergence of west nile virus in europe and continuous spread of certain flaviviruses such as dengue, yellow fever and japanese encephalitis viruses represent a global danger to public health. therefore, a rapid and accurate molecular method is required for diagnostics and epidemiological surveillance of flaviviruses.201323410000
feeding patterns of molestus and pipiens forms of culex pipiens (diptera: culicidae) in a region of high hybridization.two biological forms of the mosquito culex pipiens s.s., denoted pipiens and molestus, display behavioural differences that may affect their role as vectors of arboviruses. in this study, the feeding patterns of molestus and pipiens forms were investigated in comporta (portugal), where high levels of inter-form admixture have been recorded.201323578139
real time classification of viruses in 12 dimensions.the international committee on taxonomy of viruses authorizes and organizes the taxonomic classification of viruses. thus far, the detailed classifications for all viruses are neither complete nor free from dispute. for example, the current missing label rates in genbank are 12.1% for family label and 30.0% for genus label. using the proposed natural vector representation, all 2,044 single-segment referenced viral genomes in genbank can be embedded in [formula: see text]. unlike other approaches ...201323717598
comparative genomic and phylogenetic analysis of the first usutu virus isolate from a human patient presenting with neurological symptoms.usutu virus (usuv) is a mosquito-borne flavivirus, belonging to the japanese encephalitis antigenic complex, that circulates among mosquitoes and birds. we describe and analyze the complete genome sequence of the first usuv strain isolated from an immunocompromised patient with neuroinvasive disease. this usuv isolate showed an overall nucleotide identity of 99% and 96%, respectively, with the genomes of isolates from europe and africa. comparison of the human usuv complete polyprotein sequence ...201323741387
comparative full length genome sequence analysis of usutu virus isolates from africa.usutu virus (usuv), a flavivirus belonging to the japanese encephalitis serocomplex, was identified in south africa in 1959 and reported for the first time in europe in 2001. to date, full length genome sequences have been available only for the reference strain from south africa and a single isolate from each of austria, hungary, and italy.201323816256
recent circulation of west nile virus and potentially other closely related flaviviruses in southern france.in recent years, the number of west nile virus (wnv) cases reported in horses and humans has increased dramatically throughout the mediterranean basin. furthermore, the emergence of usutu virus (usuv) in austria in 2001, and its subsequent expansion to hungary, spain, italy, switzerland, the united kingdom, and germany, has given added cause for concern regarding the impact of the spread of flaviviruses on human and animal health in western europe. despite frequent detection of wnv and usuv case ...201323930977
[usutu virus]. 201323884536
development of a usutu virus specific real-time reverse transcription pcr assay based on sequenced strains from africa and europe.usutu virus (usuv) has been isolated in several african and european countries mainly from mosquitoes and birds. however, previous benign and two recent severe cases of human infections point out the need of a tool for the identification of usuv in human samples. a published real-time reverse transcription (rt) pcr assay for the detection of usuv in human blood or cerebrospinal fluid does not take into account the genetic variability of usuv in different geographic regions. therefore, this artic ...201424036076
first detection of co-circulation of west nile and usutu viruses in equids in the south-west of tunisia.in the last fifteen years, west nile virus (wnv) has dramatically expanded its geographic range and is now considered the most widespread arbovirus in the world. in tunisia, west nile fever (wnf) outbreaks were reported in humans in 1997, 2003 and 2012. usutu virus (usuv), which is a 'new' emerging flavivirus antigenically close to wnv, has never been reported in tunisia. a serological investigation in 284 equids was conducted in 2012 in the southern west region of the country to assess the pres ...201425065813
validation of a structural comparison of the antigenic characteristics of usutu virus and west nile virus envelope proteins.cross-reactions observed in serological assays between usutu virus (usuv), the usuv outlier subtype strain car_1969 and west nile virus (wnv) suggest that they share antigenic features amongst their structural outer proteins especially envelope (e) proteins. to investigate the molecular background of this observation, we compared the e protein sequences of seven usuv strains, usuv subtype strain car_1969 and wnv strain 2471, focusing on the binding site defined by the wnv neutralizing antibody e ...201424874193
first evidence of simultaneous occurrence of west nile virus and usutu virus neuroinvasive disease in humans in croatia during the 2013 outbreak.we report on first evidence of simultaneous occurrence of west nile virus (wnv) and usutu virus (usuv) neuroinvasive infection in humans in croatia during the transmission season 2013.201424793998
the composition of west nile virus lipid envelope unveils a role of sphingolipid metabolism in flavivirus biogenesis.west nile virus (wnv) is an emerging zoonotic mosquito-borne flavivirus responsible for outbreaks of febrile illness and meningoencephalitis. the replication of wnv takes place on virus-modified membranes from the endoplasmic reticulum of the host cell, and virions acquire their envelope by budding into this organelle. consistent with this view, the cellular biology of this pathogen is intimately linked to modifications of the intracellular membranes, and the requirement for specific lipids, suc ...201425122799
universal single-probe rt-pcr assay for diagnosis of dengue virus infections.dengue is a mosquito-borne viral disease that has become more prevalent in the last few decades. most patients are viremic when they present with symptoms, and early diagnosis of dengue is important in preventing severe clinical complications associated with this disease and also represents a key factor in differential diagnosis. here, we designed and validated a hydrolysis-probe-based one-step real-time rt-pcr assay that targets the genomes of dengue virus serotypes 1-4.201425522325
comparison of complete genome sequences of usutu virus strains detected in spain, central europe, and africa.the complete genomic sequence of usutu virus (usuv, genus flavivirus, family flaviviridae) strain mb119/06, detected in a pool of culex pipiens mosquitoes in northeastern spain (viladecans, catalonia) in 2006, was determined and analyzed. the phylogenetic relationship with all other available complete usuv genome sequences was established. the spanish sequence investigated showed the closest relationship to the usuv prototype strain sa ar 1776 isolated in south africa in 1959 (96.9% nucleotide a ...201424746182
molecular characterization of flaviviruses from field-collected mosquitoes in northwestern italy, 2011-2012.the genus flavivirus comprises several mosquito-borne species, including the zoonotic pathogens west nile and usutu virus, circulating in animals and humans in italy since 1998. due to its ecological and geographical features, piedmont is considered a risk area for flavivirus transmission. here we report the results of a flavivirus survey (detection and genetic characterization) of mosquitoes collected in piedmont in 2012 and the genetic characterization of three strains detected in 2011.201425160565
protection of a single dose west nile virus recombinant subviral particle vaccine against lineage 1 or 2 strains and analysis of the cross-reactivity with usutu virus.west nile virus (wnv) is a neurovirulent mosquito-borne flavivirus. high wnv virulence was mainly associated with lineage 1 strains, but recent outbreaks have unveiled circulation of highly virulent lineage 2 strains. co-expression of flavivirus prm and e glycoproteins drives the assembly of recombinant subviral particles (rsps) that share antigenic features with virions. mouse immunization with lineage 1 wnv rsps induced a potent humoral response against wnv with production of neutralizing anti ...201425229345
stress responses in flavivirus-infected cells: activation of unfolded protein response and autophagy.the flavivirus is a genus of rna viruses that includes multiple long known human, animal, and zoonotic pathogens such as dengue virus, yellow fever virus, west nile virus, or japanese encephalitis virus, as well as other less known viruses that represent potential threats for human and animal health such as usutu or zika viruses. flavivirus replication is based on endoplasmic reticulum-derived structures. membrane remodeling and accumulation of viral factors induce endoplasmic reticulum stress t ...201424917859
tick-borne encephalitis virus (tbev) - findings on cross reactivity and longevity of tbev antibodies in animal sera.by using animal sera as sentinels, natural tbev foci could be identified and further analyses including investigations of ticks could be initiated. however, antibody response against tbev-related flaviviruses might adversely affect the readout of such a monitoring. therefore, the cross-reactivity of the applied tbev serology test systems - enzyme linked immunosorbent assay (elisa) and virus neutralization test (vnt) - as well as the longevity of tbev antibody titres in sheep and goats were inves ...201424690234
vector competence of aedes aegypti and aedes vittatus (diptera: culicidae) from senegal and cape verde archipelago for west african lineages of chikungunya virus.to assess the risk of emergence of chikungunya virus (chikv) in west africa, vector competence of wild-type, urban, and non-urban aedes aegypti and ae. vittatus from senegal and cape verde for chikv was investigated. mosquitoes were fed orally with chikv isolates from mosquitoes (ard30237), bats (cs13-288), and humans (hd180738). after 5, 10, and 15 days of incubation following an infectious blood meal, presence of chikv rna was determined in bodies, legs/wings, and saliva using real-time revers ...201425002293
circulation of a meaban-like virus in yellow-legged gulls and seabird ticks in the western mediterranean basin.in recent years, a number of zoonotic flaviviruses have emerged worldwide, and wild birds serve as their major reservoirs. epidemiological surveys of bird populations at various geographical scales can clarify key aspects of the eco-epidemiology of these viruses. in this study, we aimed at exploring the presence of flaviviruses in the western mediterranean by sampling breeding populations of the yellow-legged gull (larus michahellis), a widely distributed, anthropophilic, and abundant seabird sp ...201424625959
rapid molecular assays for the detection of yellow fever virus in low-resource settings.yellow fever (yf) is an acute viral hemorrhagic disease transmitted by aedes mosquitoes. the causative agent, the yellow fever virus (yfv), is found in tropical and subtropical areas of south america and africa. although a vaccine is available since the 1930s, yf still causes thousands of deaths and several outbreaks have recently occurred in africa. therefore, rapid and reliable diagnostic methods easy to perform in low-resources settings could have a major impact on early detection of outbreak ...201424603874
experimental infections of wild birds with west nile virus.avian models of west nile virus (wnv) disease have become pivotal in the study of infection pathogenesis and transmission, despite the intrinsic constraints that represents this type of experimental research that needs to be conducted in biosecurity level 3 (bsl3) facilities. this review summarizes the main achievements of wnv experimental research carried out in wild birds, highlighting advantages and limitations of this model. viral and host factors that determine the infection outcome are ana ...201424531334
a model of laboratory surveillance for neuro-arbovirosis applied during 2012 in the emilia-romagna region, italy.arboviruses with neuroinvasive potential are gaining more attention due to the increased number of cases of autochthonous and imported infections in the human host. diagnosis of infection caused by these viruses in patients with central nervous system (cns) diseases is still underestimated and these infections represent an emerging threat to public health. we describe a model suitable for the laboratory surveillance of neuro-arbovirosis that was applied in the emilia-romagna region, north-easter ...201424329773
early warning of west nile virus mosquito vector: climate and land use models successfully explain phenology and abundance of culex pipiens mosquitoes in north-western italy.west nile virus (wnv) is an emerging global health threat. transmission risk is strongly related to the abundance of mosquito vectors, typically culex pipiens in europe. early-warning predictors of mosquito population dynamics would therefore help guide entomological surveillance and thereby facilitate early warnings of transmission risk.201424924622
limited efficacy of west nile virus vaccines in large falcons (falco spp.).west nile virus (wnv) can lead to fatal diseases in raptor species. unfortunately, there is no vaccine which has been designed specifically for use in breeding stocks of falcons. therefore the immunogenicity and protective capacity of two commercially available wnv vaccines, both approved for use in horses, were evaluated in large falcons. one vaccine contained adjuvanted inactivated wnv lineage 1 immunogens, while the second represented a canarypox recombinant live virus vector vaccine. the eff ...201424708385
climatic effects on mosquito abundance in mediterranean wetlands.the impact of climate change on vector-borne diseases is highly controversial. one of the principal points of debate is whether or not climate influences mosquito abundance, a key factor in disease transmission.201425030527
usutu virus in blackbirds (turdus merula), czech republic, 2011-2012.the central european lineage of usutu virus was isolated from a blackbird (turdus merula), which was found dead in the city of brno, czech republic, in 2011. the virus rna was detected in two other dead blackbirds in brno during 2012.201423095331
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