Influenza B season: low intensity with impact on mortality

Low intensity with impact on mortality - Influenza B seasonN/

Molecular epidemiology of Respiratory Syncytial Virus between 2010-2015, in Portugal

Background: Respiratory syncytial virus is one of the major causes of respiratory infection and complications in younger children and elderly. This study has, for the first time, investigated the genetic diversity of RSV A and RSV B detected since 2010, in influenza like illness (ILI) cases reported in the scope of the Portuguese Influenza Surveillance Programme (NISP). Methods: During 2010-2015, nasopharyngeal swabs (NPS) sent to the National Influenza Reference Laboratory from sentinel and non-sentinel network were tested for RSV A and RSV B by real time multiplex RT-PCR. Nucleotide sequence of a fragment of the hypervariable C-terminal region of the G protein gene and the phylogenetic analysis was performed for an half of detected RSV. Results: Over the study period were detected 114 (5.2%) RSV in 2187 tested NPS. Of these 67 (59%) were from subtype A and 47 (41%) from subtype B. Circulation of RSV preceded or was coincident with the influenza epidemic period. RSV A was predominant in each winter with exception for 2014/2015 winter when RSV B was predominantly detected. Of the RSV positive samples, 58 (51%) were successfully sequenced and genetically characterized: 26 (45%) RSV A and 32 (55%) RSV B. RSV A clustered in two genotypes. A majority (n= 22; 85%) belonged to ON1 genotype and 4 (15%) viruses belonged to NA1 genotype. Only ON1 genotype was detected after 2012/2013 season. RSV B clustered in two genotypes: a majority (n=22; 67%) belonged to BA9 genotype and 11 (33%) clustered in BA10 genotype. BA9 genotype was detected over all the study period, although BA10 was only detected in 2012/2013, and 2014/2015 seasons. Conclusion: Our study highlights the importance of RSV in ILI cases, showing a seasonal circulation each winter season during influenza epidemic. RSV accounted for 5.2% of the cases reported in the scope of influenza surveillance, assuming a huge importance in young children and older ones. Molecular data for RSVA revealed co circulation of NA1 and ON1 till 2012, and after this period ON1 was exclusively detected suggesting a strain replacement by this antigenically advantageous genotype. Globally ON1 is also predominantly detected. For RSVB subtype was observed a co circulation of the BA9 and BA10 genotypes. BA derived genotypes, first identified in 1999 in Buenos Aires are predominant in many countries since then

Antiviral susceptibility of influenza A viruses isolated between 2009-2013, in Portugal

Monitoring the influenza antiviral susceptibility had become an important issue in the recent years. The influenza A viruses are naturally susceptible to Neuraminidase inhibitors (NAI) and M2 inhibitors (adamantanes). The increased resistance to oseltamivir and adamantanes (since 2006), the availability and clinical use of influenza antivirals enhanced the need for a close monitoring for loss of susceptibility. Since 2009, at Portuguese National influenza Centre, was developed in the scope of the National Influenza Surveillance Programme a systematic monitoring of antiviral susceptibility (oseltamivir, zanamivir and adamantine). This study shows the results of antiviral susceptibility to NAI and M2 inhibitors for influenza A viruses characterized during 4 winter seasons (2009 to 2013). The phenotypic method, NA inhibition assay, considered the “gold standard” method to determine the susceptibility to oseltamivir and zanamivir, measuring the 50% inhibitory concentrations (IC50) was performed during the study period for a selection of 119 influenza A viruses: 26 and 60 AH1pdm09 from 2009/10 and 2012/13 season, respectively, and 33 AH3 viruses from 2011/12, plus 2 AH3 from the last season were also analysed. Genotypic screening methods were performed for the most common NAI inhibition-reducing substitution, H275Y in AH1pdm09 viruses (n= 340). Matrix gene sequencing, for adamantanes susceptibility surveillance, and neuraminidase gene sequencing were performed for 64 and 119 strains, respectively. In the study period all AH1pdm09 and AH3 viruses, carried the S31N substitution in the M2 protein, which confers resistance to the adamantanes to 100% of the isolates. For the AH1pdm09 the IC50 for oseltamivir ranged from 0.45 to 2.82 (2009/2010) and from 0.15 to 3.27 (2012/2013). The resistance to oseltamivir was identified in 3 AH1pdm09 viruses that carried the H275Y substitution. Two of these viruses were isolated from patients with chronic diseases treated with oseltamivir, a pregnant woman 26 years old with a fatal outcome and an 8 years old child. One of the oseltamivir-resistant virus showed an IC5o value 250-fold higher (IC5o= 502.48) comparing to the susceptible viruses. The AH1pdm09 IC5o values for zanamivir ranged from 0.32 to 3.88 (2009/2010) and from 0.29 to 2.99 (2012/2013). The median IC50 values for oseltamivir and zanamivir were stable between 2009 and 2013. None of the AH1pdm09 isolates presented the other substitutions (D119N, I223R, N295S) associated with reduced susceptibility to NAI. The AH3 viruses analysed in 2011/2012 season showed IC5o values for oseltamivir that ranged from 0.19 to 0.90 and for zanamivir from 0.39 to 0.84. The median IC50 values for oseltamivir and zanamivir were 0.41±1.55 and 0.59±1.21 respectively. In the last season 2012/2013 only two AH3 isolates were analysed and the IC5o values were in the same range of the 2011/2012 AH3 viruses. Since 2009 all the AH3 viruses are susceptible to oseltamivir and zanamivir. None of the AH3 viruses presented the amino acid substitutions known to reduce susceptibility to NAI (E119V/I, R229K, N294S and H274Y). Influenza A viruses isolated in Portugal since 2009 are resistant to adamantanes, which are no longer indicated for influenza A treatment. Otherwise, the resistance to oseltamivir was only observed in a reduced number of strains and all the viruses show susceptibility to zanamivir. The use of conventional sequence analysis and genotypic screening methods for monitoring the molecular markers of antiviral resistance in influenza A virus provides a valuable tool for an early detection of antiviral resistant strains

Antiviral resistance: influenza B

Currently circulating influenza viruses are resistant to adamantanes and except for a low number of sporadic cases most are sensitive to neuraminidase inhibitors (NI). Adamantanes are ineffective against influenza B viruses and although NI-resistant influenza B viruses have been rarely reported, recently in the United States was identified one cluster of influenza B viruses with reduced susceptibility to NI and with the I221V substitution in the active site of the neuraminidase. Despite the low prevalence of oseltamivir-resistant influenza viruses, the constant evolution of influenza requires the monitoring of antiviral resistance among these viruses in the community. This is very important for the clinical management of severe influenza cases as to the detection of novel genetic markers associated with antiviral resistance. This study reports the antiviral susceptibility to neuraminidase inhibitors of influenza B viruses isolated in Portugal during the 2010/2011, 2011/2011 and 2012/2013 seasons. Over the period of 3 influenza seasons, 146 influenza B viral strains were selected for phenotypic fluorescent assays in order to assess their susceptibility to NI, oseltamivir and zanamivir. For this purpose, was determined the NI concentration required to inhibit 50% of each influenza virus neuraminidase activity (IC50). The IC50 baseline of influenza B viruses was calculated for both oseltamivir and zanamivir using the Robust Excel programme. The neuraminidase gene segments were also monitored for the presence of the main molecular markers, associated with the resistance to neuraminidase inhibitors in influenza B viruses. All analysed influenza B strains proved to be susceptible to oseltamivir and zanamivir. In the 2010/2011 season the determined IC50 values ranged from 0 to 70 nM for oseltamivir and from 0 to 11nM to zanamivir. The zanamivir IC50 median value was about 8-fold lower than oseltamivir IC50 median value. Statistical analysis revealed the presence of one outlier (B/Lisboa/13/2010, 71.08nM) for oseltamivir (2-fold reduction in susceptibility) and four minor outliers (B/Lisboa/15/2010: 9.52 nM; B/Lisboa/19/2010: 9.25 nM; B/Lisboa/51/2010: 10.94 nM and B/Lisboa/53/2010: 9.62 nM) for zanamivir (3-fold reduction in susceptibility) comparing to the median IC50 value. During the 2011/2012 season IC50 values ranged from 23.0 to 38.38 nM (oseltamivir) and from 2.97 to 9.07nM (zanamivir). This season were not found minor or major outliers. The IC50 values obtained in 2012/2013 ranged from 7.91 to 84.84 nM (oseltamivir) and from 1.48 to 5.88 nM (zanamivir). During this last season were found 6 minor and 3 major outliers for oseltamivir and 13 minor outliers for zanamivir. Along the three seasons, the median IC50 values for both NI were higher among the B/Victoria than B/Yamagata viruses. None of the actually known mutations associated with resistance of influenza B viruses to NI was found in the neuraminidase gene (R150K, D197E/N/Y, I221T/V, N294S, R374K and G407S). Influenza B viruses isolated in Portugal during the last three seasons were susceptible to the neuraminidase inhibitors. Portuguese influenza B strains revealed higher susceptibility to zanamivir than to oseltamivir, as observed in other countries. The oseltamivir IC50 values were also different between viruses from the B/Victoria and B/Yamagata lineages, however this was not statistically demonstrated due to the small number of analysed viruses. Among B/Yamagata strains the oseltamivir and zanamivir IC50 values were higher in 2011/2012 than in 2010/2011 and 2012/2013 seasons

Performance of ECDC ILI case definition and ICPC R80 code for influenza surveillance

In this study we took advantage of the Portuguese Influenza Surveillance Systems (ISS) database to assess and compare the performance of the two main case definitions used in Portugal: the European Centre for Disease Prevention and Control (ECDC) ILI case definition and the International Classification of Primary Care (ICPC) R80 code.N/

Avaliação de um projeto de educação ambiental via internet: o programar educ@r

El objetivo de esta investigación fue la identificación de algunos aspectos que facilitaran y/o dificultaran la implantación y una mejor adecuación de un projecto de Educación Ambiental a distancia via Rede Internet, denominado "Projeto Educ@r" coordinado por la CDCC/USP/São Carlos (Brasil).The aim of this research was the identification of some aspects that facilitate and/or make difficult the implantation and better adjustment of an project of Environmental Education at distance by Internet nominated Educ@r Project, which was coordinate by CDCC/University of São Paulo/São Carlos (Brazil)

Programa Nacional de Vigilância da Gripe: relatório da época 2010/2011

A época de vigilância da gripe de 2010/2011 caracteriza-se por ser a época posterior à primeira pandemia do século XXI, que esteve associada à circulação de um novo subtipo de vírus influenza, o vírus pandémico A(H1N1)pdm09. Nesta época verificou-se um pico de atividade gripal entre as semanas 50/2010 e 5/2011, com a duração de 8 semanas. Durante este período a taxa de incidência atingiu o valor máximo de 121,12 casos por 100 000 habitantes, na semana 52, sendo a atividade gripal considerada como alta/moderada. Do ponto de vista virológico a época teve uma apresentação bifásica, primeiramente dominada pelos vírus influenza do tipo B da linhagem Victoria, e mais tarde pela circulação dos vírus influenza A(H1)pdm09, sendo estes últimos os detetados num maior números de casos (55,7%). A deteção dos vírus influenza do subtipo A(H3) e do tipo B da linhagem Yamagata foi esporádica, e os vírus influenza A(H1) sazonais não foram detetados. Os vírus estudados foram, antigénica e geneticamente, semelhantes às estirpes incluídas na composição da vacina antigripal disponível para a época. Durante a época de 2010/2011, a maior percentagem de casos de gripe (70,4%) foi detetada no grupo das crianças em idade escolar (5-14 anos). Nestas, a doença esteve particularmente associada à infeção com vírus influenza do tipo B, enquanto que a infeção com vírus influenza do tipo A ocorreu, maioritariamente, na população adulta. Na época em estudo ocorreu uma redução na cobertura da vacina antigripal sazonal, na população geral em todos os grupos etários à exceção dos 45-64 anos e em todos os grupos que declararam sofrer de doenças crónicas, à exceção dos diabéticos. Apesar de nenhuma destas diferenças ser estatisticamente significativa, a sua observação concomitante deverá ser considerada como um alerta, incentivando assim o reforço da campanha de vacinação nos grupos alvos prioritários para a vacinação antigripal