Virus In Spain Research Articles

Ten Years of Global Evolution of the Human Respiratory Syncytial Virus BA Genotype with a 60-Nucleotide Duplication in the G Protein Gene

The emergence of natural isolates of human respiratory syncytial virus group B (HRSV-B) with a 60-nucleotide (nt) duplication in the G protein gene in Buenos Aires, Argentina, in 1999 (A. Trento et al., J. Gen. Virol. 84:3115-3120, 2003) and their dissemination worldwide allowed us to use the duplicated segment as a natural tag to examine in detail the evolution of HRSV during propagation in its natural host. Viruses with the duplicated segment were all clustered in a new genotype, named BA (A. Trento et al., J. Virol. 80:975-984, 2006). To obtain information about the prevalence of these viruses in Spain, we tested for the presence of the duplicated segment in positive HRSV-B clinical samples collected at the Severo Ochoa Hospital (Madrid) during 12 consecutive epidemics (1996-1997 to 2007-2008). Viruses with the 60-nt duplication were found in 61 samples, with a high prevalence relative to the rest of B genotypes in the most recent seasons. Global phylogenetic and demographic analysis of all G sequences containing the duplication, collected across five continents up until April 2009, revealed that the prevalence of the BA genotype increased gradually until 2004-2005, despite its rapid dissemination worldwide. After that date and coinciding with a bottleneck effect on the population size, a relatively new BA lineage (BA-IV) replaced all other group B viruses, suggesting further adaptation of the BA genotype to its natural host.

Detection of low pathogenic avian influenza viruses in wild birds in Castilla-La Mancha (south central Spain)

The Iberian Peninsula is located along the East Atlantic and Black Sea/Mediterranean flyways and is the third ranking European country as wintering quarter for wild migrating birds after Turkey and Rumania. For these reasons, Spanish wetlands are of importance in AIV surveillance, and of great interest for the study of the epidemiology of LPAIV under Mediterranean climate conditions. Nevertheless, information on prevalence of LPAIV viruses in Spain is still scarce and is restricted to two serological surveys carried out in the south of the country during 1990 and 1994 and one virological study performed recently in North East Spain. In the present study we analysed the prevalence of AIV circulating in wild birds in continental wetlands in central Spain and determined temporal, spatial and species variation. Real time RTPCR was performed on 1435 faecal samples and cloacal swabs from 54 species. An overall AIV prevalence of 2.6% was detected with a peak during November and December, when thousands of migrating wild birds arrive to Spain for wintering. Highest prevalence rates were detected in Phoenicopteriformes and Anseriformes. AIV prevalence obtained from cloacal swabs and fresh faeces did not vary significantly, which supports faecal sampling as an appropriate method for large scale LPAIV surveillance programs. Viral culture was achieved in samples obtained from two Mallards and a White stork, in which subtypes H7N9 and H11N9, respectively, were identified. Our results reflect a similar scenario in AIV epidemiology in small continental wetlands as compared to large coastal humid areas in Europe and underline the importance of including species such as flamingos and storks in surveillance programs, since their role in AIV ecology in these areas could be more important than previously considered.

First Report of Pepper as a Natural Host for Pelargonium zonate spot virus in Spain.

Pelargonium zonate spot virus (PZSV) was first reported on Pelargonium zonale (L.) L'Hér. ex Aiton and later on tomato in Italy, Spain, France (1), and the United States (2). In Spain, PZSV was first detected in 1996 in tomato plants of cv. Royesta from greenhouses in Zaragoza Province (3) and subsequently in tomato in the Catalonia and Navarra areas. In April 2006, symptoms of PZSV were found at high incidence on tomato in a greenhouse in Huesca, Aragón (northeastern Spain). Randomly distributed pepper plants (Capsicum annuum L.) of cv. Estilo F1 growing in the same greenhouse showed severe foliar chlorotic ringspots and line patterns similar to those observed in tomato. Samples from symptomatic peppers and tomatoes and one asymptomatic weed of Rubia tinctorum L. tested positive by double-antibody sandwich (DAS)-ELISA using polyclonal antibodies against PZSV (Agdia Inc., Elkhart, IN and DSMZ, Braunschweig, Germany) as did a Spanish PZSV isolate used as a positive control (3). Sap extracts from two tomatoes, three peppers, and the single R. tinctorum plant were mechanically inoculated to 22 indicator species, including pepper and tomato. On 17 of 22 species inoculated, sap from symptomatic tomatoes and peppers elicited local or systemic symptoms similar to those reported earlier for PZSV isolates (3). Systemic symptoms were mainly mosaic, chlorotic, and necrotic line patterns and ringspots on leaves of most indicator species, closely resembling those observed on the greenhouse pepper and tomato plants. Symptoms on inoculated tomatoes also included stem necrosis and death. Reactions of indicator species did not indicate the presence of any other pepper- or tomato-infecting viruses. Both field infected and mechanically inoculated plants of pepper cvs. Yolo Wonder and Doux des Landes were maintained in the greenhouse until the development of fruit symptoms. Only fruits of cv. Yolo Wonder showed dark green and slightly depressed circles on their surface. Local and systemic infection by PZSV was confirmed by DAS-ELISA in most inoculated plants. Total RNA from leaves of field or inoculated plants was used as template for amplification by reverse transcription (RT)-PCR with primers R3-F and R3-R that are specific for the PZSV 3a gene (2), and amplicons were sequenced directly. The sequences of 697 nt from pepper and tomato isolates from the same greenhouse were identical (GenBank Accession Nos. CQ178217 and CQ178216, respectively) and had 96.1% identity to nucleotides 384 to 1,080 in PZSV RNA-3 (NC_003651). Our results confirm the natural infection of pepper plants in Huesca by PZSV. To our knowledge, this is the first report of pepper as a natural host for PZSV, a significant finding considering the potential risks of PZSV dispersion whenever tomato and pepper coexist, particularly in greenhouses and nurseries.

Molecular characterisation and recent evolution of myxoma virus in Spain

Myxoma virus and the European rabbit are one of the best-studied examples of coevolution of pathogen virulence and host resistance. Since the introduction of the virus in Spain in 1953, a decrease in its virulence has been observed; however, most strains are still considered highly virulent. To determine whether this attenuation is due to molecular differences, and to characterise the field strains in Spain and the genetic changes that have occurred since the introduction of the virus, we analysed 7,741 bp in 97 virus samples from 12 localities. We found an extremely low genetic variability and an absence of a geographic structure. We defined 35 haplotypes, none of which were identical to the original Lausanne strain. Three genetic groups were determined and were found to occur at different frequencies in different locations. Overall, virus evolution deviated from neutrality and did not conform to a strict molecular clock, probably due to the existence of a strong selective pressure that acts differently across the viral genome.

Hepatitis C and hepatitis B-related mortality in Spain

Although hepatitis C and hepatitis B virus (HCV/HBV) infections are an important health problem worldwide, their burden of disease (BoD) taking into account their chronic consequences, cirrhosis, and hepatocellular carcinoma (HCC), is still unknown. Our aim was to assess the total number of deaths attributable to these viruses in Spain and the years of life lost, first component of the disability adjusted life years, a comparative index of BoD. We selected the International Classification of Diseases-Tenth Revision specific codes related to HCV/HBV. For unspecified cirrhosis and HCC, the attributable fraction of mortality was assessed in two steps: literature review and expert panel. Deaths in Spain in 2000 were obtained from the National Statistics Institute. Years of life lost were calculated using the estimated mortality and life expectancies (Princeton Model Life Table). HCV could have caused around 70% of HCC deaths and 50% of cirrhosis mortality in Spain in 2000 (60% HCC, 40% cirrhosis with HCV lower estimate). For HBV these proportions are 10 and 13%, respectively. We estimated 4342 HCV-related deaths and 877 HBV-related deaths in Spain in 2000, globally 1.5% of total deaths in Spain that year. Mortality by cirrhosis and HCC represented most of these viral-related deaths. Attributable mortality in AIDS patients was also estimated. HCV leads the list of infectious disease-related mortality in Spain in 2000, doubling the AIDS mortality even if lower HCV attributable fractions are considered. Exclusion of cirrhosis and HCC-related mortality severely underestimates the BoD attributable to HCV/HBV. Improving early diagnosis and access to treatment could have an important impact on mortality because of hepatitis virus in the next decades.

New influenza A(H1N1) virus infections in Spain, April-May 2009

An outbreak of infections with a new influenza A(H1N1) virus that was first detected in the United States and Mexico is currently ongoing worldwide. This report describes the initial epidemiological actions and outbreak investigation of the first 98 laboratory confirmed cases of infection with this new virus in Spain.

Tomato infectious chlorosis virus

<i>Tomato infectious chlorosis virus</i>

Ecological correlates of bluetongue virus in Spain: Predicted spatial occurrence and its relationship with the observed abundance of the potential Culicoides spp. vector

Using data from bluetongue (BT) outbreaks caused by viral serotype 4 (BTV-4) in Spain during 2004-2005, a predictive model for BTV-4 occurrence in peninsular Spain was developed. An autologistic regression model was employed to estimate the relationships between BTV-4 presence and bioclimatic-related and host-availability-related variables. In addition, the observed abundances of the main potential Culicoides vectors during 2004-2005, namely Culicoides imicola, Culicoides obsoletus group, and species of the Culicoides pulicaris group, were compared between BTV-4 presence/absence areas predicted by the model. BTV-4 occurrence was mainly explained by bioclimatic variables, although a consideration of host-availability variables led to improved fit of the model. The area of BTV-4 presence predicted by the model largely resembled the core distribution area of C. imicola, and this species was the most abundant Culicoides spp. in predicted BTV-4 presence areas. The results suggest that the spatial expansion of BTV-4 took place only as far as those areas in which C. imicola populations efficiently transmitted the virus.

Molecular epidemiology and evolution of avian infectious bronchitis virus in Spain over a fourteen-year period

An in-depth molecular study of infectious bronchitis viruses (IBV) with particular interest in evolutionary aspects of IBV in Spain was carried out in the present study based on the S1 gene molecular characterization of twenty-six Spanish strains isolated over a fourteen-year period. Four genotypes were identified based on S1 gene sequence analyses and phylogenetic studies. A drastic virus population shift was demonstrated along time and the novel Italy 02 serotype was shown to have displaced the previous predominant serotype 4/91 in the field. Detailed analyses of synonymous to non-synonymous ratio of the S1 gene sequences of this new serotype Italy 02 suggested positive selection pressures might have contributed to the successful establishment of Italy 02 serotype in our country. In addition, differences on the fitness abilities of new emergent genotypes were indicated. Furthermore, intergenic sequences (IGs)-like motifs within S1 gene sequences of IBV isolates were suggested to enhance the recombination abilities of certain serotypes.

First case of highly pathogenic H5N1 avian influenza virus in Spain

BackgroundThe H5N1 strain of avian influenza virus has been involved in severe mortality in domestic poultry, and has also been found in different species of wildlife in Europe. The Basque Country avian influenza surveillance program began sample collection and processing the fall of 2005.ResultsHere we report the first confirmation of the presence of highly pathogenic H5N1 strain in a Great Crested Grebe (Podiceps cristatus) found dead in a pond near Vitoria in the Basque Country on the North of Spain. Regarding the survey for generic influenza type A virus, we have obtained positive results in about 8% of more that 3500 birds examined.ConclusionWe think that the self-limiting nature of our finding and others proves that certain regions have ecological, geographical and climatological features that make it difficult for the H5N1 virus to spread [1] and cause disease at least in the large scale scenario that has been worrying human and animal health authorities during the last years.

High incidence of human bocavirus infection in children in Spain

Background The newly identified human bocavirus (HBoV), a member of the Parvoviridae family, has been associated to low respiratory tract infections in young children. Objectives To present the epidemiological profile and the main clinical characteristics showed by children infected with this virus in Spain. Study design We have studied the incidence of HBoV and other 15 respiratory viruses in 917 nasopharyngeal aspirates taken from 730 infants and children under age of 14 with acute lower respiratory tract infection from September-04 to August-06. Results HBoV was detected in 123 samples (13.4%) showing a seasonal distribution with November and December as the peak months. Out of the 558 samples which rendered a positive result for at least one of the virus tested, HBoV (22%) ranked fourth behind respiratory syncytial virus (181, 32%), adenoviruses (155, 28%) and rhinoviruses (136, 24%). Co-infections with HBoV and other respiratory viruses were detected in 74 out of 123 HBoV-positive specimens (60%). In addition, HBoV was also found in stool and, for the first time, in urine samples. Conclusions Results obtained provide further evidence that HBoV is involved in acute lower respiratory tract infections. HBoV-associated disease should not be limited to the respiratory tract.

Risk Assessment Applied to Spain's Prevention Strategy Against Highly Pathogenic Avian Influenza Virus H5N1

Notifiable avian influenza (NAI) had never been reported in Spain, until July 2006 when a dead Great Crested Grebe (Podiceps cristatus) was found positive to the highly pathogenic H5N1 subtype as part of the active wild bird surveillance plan. The current program of the Spanish Ministry of Agriculture, Fisheries, and Food (MAPA)'s strategic preventive plan against NAI is divided in the following parts: identification of risk areas and risk wild bird species, increased biosecurity measures, early detection of infection with surveillance intensification and development of rapid diagnostic tests, and other policies, which include continuing education and training to ensure early detection of the disease. In 2003 an active surveillance plan was introduced for domestic fowl; the plan was extended to wild birds in 2004. A total of 18,780 samples in poultry and 3687 samples in wild birds had been analyzed through December 2005 to detect the presence and spread of avian influenza subtypes H5 and H7. In the present work we suggest some contributions to be implemented in MAPA's action plan: 1) the identification of risks because of migratory birds, within the risk assessment of the introduction of NAI virus in Spain and 2) an interactive digital simulator of the disease developed for continuing education and training.

Prevalence of Antibodies against Rubella Virus in Spain

The importance acquired by the strategy of anticipating rubella revacunation needs no emphasis1. Recommendations by experts committees and actualization of the prevailing childhood immunization schedules are unanimous when it comes to including the above-mentioned strategy2. In this context, seroepidemiologic survey studies allow the assesment of humoral immunologic response against viral structural antigens3,4 despite the assumed potential bias attached to their design. Being aware of the importance of describing rubella seropositivity real status in our setting we decided to document such situation in children under theoretically proper immunization schedules. The present contribution relies on data from a cross-sectional study carried out between 1999 and 2000 in children from the autonomous region of Spain, Castilla y Leon, (the largest region in the European Economic Community). The chosen framework was restricted to serum samples received in a Microbiology Laboratory of a University Hospital with the request of analyzing infectious markers other than rubella antibodies. According to demographic features a double stratification was made and we evaluated 323 children whose age was between 1 and 5 years and 1166 from 6 to 14 years of age. All samples were alicuoted and frozen (-20oC) until the moment of processing. Antibodies to proteic antigens of rubella viral envelope were determined by means of an indirect enzymeimmunoassay (EIA) (Bio-Whittaker, USA). Results were validated according to manufacturer's instructions and samples that showed a neat absorbance greater the cut-off value plus 15% were considered positive. Our findings reveal that 309 of the investigated samples belonging to children from 1 to 5 years of age (95.7%) had antibodies to rubella virus (95% CI, 93.2% to 98.2%), being the rest of them seronegative at the moment of our study. In the age group ranging from 6 to 14 years the prevalence of antibodies was 90.5% (1055/1166) (95% CI, 88.6% to 92.5%) which resulted lower to that found in the previous group and the difference was statistically significant (p=0.003). This fact reflects a decrease of 5.2% in the seropositivity percentage of the second age group. An additional finding was that when we investigated the 6 to 14 years old group in terms of gender differences, girls (568) reached significantly higher percentages of seroprotection than boys (521) did (94.4 vs 87.1%, p=0.000). Conscious of the caution required in this kind of studies and assuming the internal validity for the evaluated population, our results point out to a age dependent loss of seroprotection. Among the potential causes that maintain this fact we may denote on the one hand the differences in vaccine coverage reached by both groups of children, just as Davidkina et al.5 have recently reported, and on the other hand limitations inherent in the vaccination itself6,7. Moreover, it is of course true that in our country, as well as in all developed countries, systematic vaccination of girls before puberty yields high efficiency, in order to prevent consequences derived from a potential primary infection during pregnancy8,9, all the same it is also certain that we still attend to the presence of small proportions of unprotected persons. The efficiency of new strategies of anticipating combined vaccines needs to be evaluated, and seroepidemiological studies seem to be a good tool for such

West Nile virus in Spain: Report of the first diagnosed case (in Spain) in a human with aseptic meningitis

We report the first case of illness caused by West Nile virus (WNV) so far diagnosed in Spain. A 21-y-old male presented with clinical and biological signs compatible with viral meningitis. Acute and convalescent serum samples showed IgM and IgG positivity for WNV. These results were confirmed by microneutralization assays.

Hepatitis C en España

Spain has a medium endemicity of hepatitic C infection among central Europe countries and Italy. Prevalence of anti-HCV varies among regions and it ranges from 1.6 to 2.6%, which means that there may be between 480,000 and 760,000 people infected with hepatitis C virus in Spain. The prevalence is very low in people under 20 years of age and it increases from age 30 years. Prisoners and drug addicts have the highest infectious rates, between 40 and 98%. Some populations of immigrants also have a high prevalence of HCV infection, especially people from Asia and sub-Saharan countries, whereas people from Latin America have rates lower than those in the autochtones population. Spanish people with chronic hepatitis C were mainly infected via blood transfusions, IV drug use, or during some medical and surgical hospitalization. The reduction in the use of IV drugs and the programs of needle sharing, as well as the eradication of post-transfusional hepatitis, have led to a progressive reduction in the incidence of new infections (from 6.8 per 100,000 in-habitants in 1997 to 2.3 in 2003). Preliminary data suggest that an important rate of new hepatitis C cases owe to nosocomial transmission. Transmission is almost exclusively vertical in children. In spite of a two-third reduction of incident cases of hepatitis C in Spain in last few years, it is foreseeable that the number of patients with advanced HCV liver disease attended in the health-care system will increase in forthcoming years. This is due to the fact that many, still undiagnosed patients will be likely recognized for the first time as a result of some complication of the disease. All efforts to increase the screening of hidden cases of hepatitis C in primary health-care centers, allowing a prompt treatment before an advanced stage, will have a beneficial impact both in economic and social terms.

Cucumber vein yellowing ipomovirus

<i>Cucumber vein yellowing ipomovirus</i>

Toscana Virus in Spain

Toscana virus (TOSV, Phlebovirus, family Bunyaviridae) infection is one of the most prevalent arboviruses in Spain. Within the objectives of a multidisciplinary network, a study on the epidemiology of TOSV was conducted in Granada, in southern Spain. The overall seroprevalence rate was 24.9%, significantly increasing with age. TOSV was detected in 3 of 103 sandfly pools by viral culture or reverse transcription-polymerase chain reaction from a region of the L gene. Nucleotide sequence homology was 99%-100% in TOSV from vectors and patients and 80%-81% compared to the Italian strain ISS Phl.3. Sequencing of the N gene of TOSV isolates from patients and vectors indicated 87%-88% and 100% homology at the nucleotide and amino acid levels, respectively, compared to the Italian strain. These findings demonstrate the circulation of at least 2 different lineages of TOSV in the Mediterranean basin, the Italian lineage and the Spanish lineage.

Molecular epidemiology of hepatitis B virus in Spain: Identification of viral genotypes and prediction of antigenic subtypes by limited sequencing

The hepatitis B virus (HBV) genotypes were studied by a line probe assay (LiPA) and by direct sequencing of a 339 nucleotide fragment from the S region of the viral genome in samples from 269 carriers living in Spain, either native to Spain (231) or immigrants from Africa, Asia, and Eastern Europe (38). The sequences were also used to predict the HBV surface antigen (HBsAg) subtype on the basis of the amino acids specified at selected positions of the HBsAg molecule. Agreement between the two genotyping methods was found in most cases (98.1%) and a HBV genotype could be assigned to all samples. The viral groups D/ayw2 (30.1%), D/ayw3 (28.6%), and A/adw2 (21.2%) were prevalent, with an additional participation of the groups D/ayw4 (4.8%), F/adw4q- (1.9%), A/ayw1 (1.9%), and D/adw3 (0.7%), all of them present among the autochthonous carriers. Strains from genotypes B and C were found exclusively among Chinese immigrants. Genotype E strains were found in immigrants from Central Africa and in one patient native of Spain. Point mutations leading to amino acid changes of residues involved in the expression of the HBsAg subtype determinants were found in 12 samples (4.5%). Some mutations would predict the putative novel genotype-subtype associations A/adw4q+, A/ayr, D/ayr, and E/ayw1, while others would suggest the loss of subtype-specific determinants. The finding of HBV strains characteristic for Africa among the autochthonous carriers confirms the emergence of African HBV strains in Spain.

Preliminary Evaluation of the Sensitivity of Alemow Rootstock to Citrus tristeza virus in Spain

Author(s): Piquer, J.; Pina, J. A.; Perez-Panades, J.; Carbonell, E. A.; Guerra, J.; Moreno, P.

HIV-1 subtype J recombinant viruses in Spain.

Genetic characterization of HIV-1 was carried out in two women who originated in Cameroon and Equatorial Guinea, respectively, and who were diagnosed more recently as HIV-1 seropositive in Madrid, Spain. Phylogenetic studies showed that the protease-encoding gene from both individuals clustered with subtype J sequences with a high bootstrap. However, env sequences clustered with subtypes A and C, respectively. This work represents the first characterization of HIV-1 containing subtype J-like genomic regions in Spain.