Made available in DSpace on 2014-08-20T14:37:55Z (GMT). No. of bitstreams: 1 dissertacao_leandro_ribas.pdf: 533423 bytes, checksum: 051e6999778a1a91a3b217f25f75e25b (MD5) Previous issue date: 2008-03-10 / The present study has the objective to evaluate the epidemiologic factors related to respiratory diseases in thoroughbred foals placed in four stud farms around the city of Bagé/RS region, Brazil. Apart from the collection of data linked to breeding management practices, the evaluations consisted of the diary clinical control and complementary examinations for diagnosis using secretions collected from the respiratory tract of the foals affected with respiratory diseases. Among the 349 monitored foals until their 180 days of life, the incidence of 9,5% (33) of respiratory cases with 0,57% (2) of mortality was recorded. The concentration of cases was higher in months of summer, the 120 and 180 days old foals were more susceptible. The occurrence of the diseases was influenced by the breeding system and management practices that increased the environmental contamination potential, and also by the contact among the foals. No clinic case was related to the equine influenza virus (EIV) and to the equine herpesvirus (EHV) pathogens. Among the isolated pathogens, the highest frequency was for Streptococcus equi (57%), followed by Rhodococcus equi (17%) and this one was responsible for 50% of lethality. The obtained results suggest that connected factors to the management in the breeding of thoroughbred equine are going to contribute decisively for the manifestation of the respiratory disease and alert for the high morbidity caused by the equine adenitis and the high lethality related to Rhodococcus equi pathogens. / O presente estudo teve por objetivo avaliar fatores epidemiológicos associados com doenças respiratórias em potros Puro Sangue Inglês (PSI) alojados em 4 propriedades na região da cidade de Bagé-RS, Brasil. Além da coleta de dados ligados ao manejo de criação, as avaliações consistiram no monitoramento clínico diário e exames complementares de diagnóstico a partir de secreções colhidas do trato respiratório de potros com enfermidade respiratória. Entre os 349 potros monitorados até os 180 dias de vida, pôde-se registrar a incidência de 9,5% (33) de casos respiratórios, com mortalidade de 0,57% (2). A concentração de casos foi mais elevada nos meses de verão e potros com idade entre 120 e 180 dias foram mais suscetíveis. A ocorrência de doenças foi influenciada pelo sistema de criação e práticas de manejo que aumentaram o potencial de contaminação do ambiente e o contato entre potros. Nenhum caso clínico foi relacionado ao vírus da influenza (EIV) e ao herpesvírus eqüino (EHV). Entre os isolados, a maior freqüência foi de Streptococcus equi (57%), seguido do Rhodococcus equi (17%), este responsável pela letalidade de 50%. Os resultados sugerem que fatores ligados ao manejo na criação de eqüinos PSI parecem contribuir decisivamente para a manifestação da doença respiratória e alertam para a elevada morbidade causada pela adenite eqüina e a alta letalidade associada à rodococose entre potros jovens (>180 dias).

Contexte : Les infections respiratoires aiguës (ARI) sont reconnues comme une cause importante de morbidité, de mortalité et d'hospitalisation chez les enfants dans les pays en développement. Le virus respiratoire syncytial humain (HRSV) est l’agent étiologique principal de maladie sévère des voies respiratoires basses chez les nourrissons, les jeunes enfants et les personnes âgées. Identifié en 2001, le Metapneumovirus humain (HMPV) est un nouveau paramyxovirus. Les études ont montré la cocirculation des sous groupes de ces deux virus avec la domination de l’un des sous groupes selon les zones géographiques et selon les années. Les données restent cependant limitées dans les pays de l’Afrique subsaharienne, sur la prévalence, la saisonnalité et la caractérisation génétique de ces deux virus respiratoires. Au Cameroun, ces deux virus ont été décrits seulement une seule fois (5,7 et 5% pour HRSV et HMPV respectivement) chez des patients présentant des syndromes grippaux en 2012. Objectif : Cette étude rapporte la prévalence, la saisonnalité et la variabilité génétique des souches HRSV et HMPV chez des enfants camerounais pendant 3 saisons épidémiques consécutives (de Septembre 2011 à Octobre 2014). Par ailleurs, la diversité génétique d’autres virus respiratoires détectés au cours de ce travail est présentée comme objectif secondaire.Méthodes : Une surveillance prospective a été menée pour identifier les enfants hospitalisés et ambulatoires âgés de moins de 15 ans présentant des symptômes respiratoires ≤ 5 jours. Les échantillons nasopharyngés ont été testés pour 17 virus respiratoires en utilisant une réaction multiplex de polymérisation en chaîne. La distribution virale et les données démographiques ont été analysées statistiquement. Les échantillons positifs du HRSV et HMPV ont été amplifiés par polymérisation en chaine semi nichée puis séquencés partiellement au niveau du gène G. Des analyses phylogénétiques ont été effectuées sur les séquences nucléotidiques et protéiques partielles du gène G.Résultats : De septembre 2011 à octobre 2014, 822 enfants âgés de moins de 15 ans ont été inscrits dans l’étude. Au moins un virus a été identifié chez chacun des 72,6% (597/822) d'enfants, dont 31,7% (189/597) étaient des codétections; 28,5% (226/822) étaient positifs pour l'adénovirus humain, 21,4% (176/822) pour le virus Influenza, 15,5% (127.822) pour le rhinovirus/entérovirus, 9,4% (77/822) pour le bocavirus, 9% (74/822) pour le HRSV, 8,2% (67/822) pour les coronavirus humain, 6,2% (50/822) pour le parainfluenzavirus humain et 3,9% (32/822) pour le HMPV. L’infection HRSV était plus fréquente chez les enfants de moins de 2 ans (70,3% ; 52/74) et chez les participants hospitalisés (70,3% ; 52/74). Alors que le HRSV a montré un profil saisonnier avec une circulation de septembre à décembre, des cas sporadiques de HMPV ont été détectés tout au long de l'année. HRSV-A (19,1%, 9/47) et HRSV-B (17% ; 8/47) ont été observés relativement à la même fréquence avec (63,8% ; 30/47) de cas en codétection HRSV-A/HRSV-B alors que HMPV-A (71,4% ; 10/14) était majoritaire comparé à HMPV-B (28,6 ; 4/14). L'analyse phylogénétique a révélé que les souches HRSV de l’étude sont groupées au sein du sous groupe NA-1 (pour HRSV-A) et BA-9 (pour HRSV-B). Les souches HMPV camerounaises sont groupés parmi les membres du génotype A2b (pour HMPV-A), B1 et B2 (pour HMPV-B).Conclusion : Cette étude suggère qu’environ 70% des ARI enregistrés chez des enfants au Cameroun sont causés par des virus. La présente étude est également le premier rapport sur la variabilité génétique du gène G des souches de HRSV et HMPV dans la région. Bien que ce travail comble partiellement certaines lacunes d’informations, des études supplémentaires sont requises pour une clarification de l’épidémiologie moléculaire et du mode d’évolution des virus respiratoires présents en Afrique subsaharienne en général et plus singulièrement au Cameroun. / Background: Acute respiratory infections (ARI) are recognized as an important cause of morbidity, mortality and hospitalization among children in developing countries. Human respiratory syncytial virus (HRSV) is the main cause of severe lower respiratory tract disease in infants, young children and the elderly. Identified in 2001, Human Metapneumovirus (HMPV) is a new paramyxovirus. Studies have shown the co-circulation of the subgroups of these two viruses with domination of one of the sub-groups according to the geographical zones and according of years. These two viruses encode two major surface glycoproteins, the highly conserved fusion F protein and the highly variable attachment G protein. Data are still limited in sub-Saharan African countries on prevalence, seasonality and genetic characterization of these two respiratory viruses. In Cameroon, these two viruses have been described only once (5.7 and 5% for HRSV and HMPV respectively) in patients with influenza-like illness in 2012.Objective: This study reports the prevalence, seasonality and the genetic variability of HRSV and HMPV strains in Cameroonian children for 3 consecutive epidemic seasons (September 2011-October 2014). Moreover, the genetic diversity of other respiratory viruses detected during this work is presented as a secondary objective.Methods: A prospective surveillance was conducted to identify inpatient and outpatient children less than 15 years with respiratory symptoms ≤ 5 days. The nasopharyngeal samples were tested for 17 respiratory viruses using a multiplex polymerase chain reaction. Viral distribution and demographic data were analyzed statistically. Positive samples for HRSV and HMPV were amplified by semi-nested polymerize chain reaction and then partially sequenced at the G gene. Phylogenetic analyzes were performed on the partial nucleotide and protein sequences of the G gene.Results: From September 2011 to October 2014, 822 children under 15 years were enrolled in the study. At least one virus was identified in each of 72.6% (577/822) of children, 31.7% (189/597) of whom were co-detections; 28.5% (226/822) were positive for human adenovirus, 21.4% (176/822) for influenza virus, 15.5% (127.822) for rhinovirus/enterovirus, 9.4% (77/822) for bocavirus, 9% (74/822) for HRSV, 8.2% (67/822) for human coronavirus, 6.2% (50/822) for human parainfluenzavirus, and 3.9% (32/822) for HMPV. HRSV infection was more frequent in children under 2 years (70.3%, 52/74) and hospitalized participants (70.3%, 52/74). While HRSV showed a seasonal pattern with circulation from September to December, sporadic cases of HMPV were detected throughout the year. HRSV-A (19.1%, 9/47) and HRSV-B (17%; 8/47) were observed relatively at the same frequency with (63.8%, 30/47) codetections of HRSV-A/HRSV-B. HMPV-A (71.4%; 10/14) was predominant compared to HMPV-B (28.6; 4/14). Phylogenetic analysis revealed that the HRSV strains of the study are grouped within subgroup NA-1 (for HRSV-A) and BA-9 (for HRSV-B). Cameroonian HMPV strains are grouped among the members of genotype A2b (for HMPV-A), B1 and B2 (for HMPV-B).Conclusion: This study suggests that about 70% of ARI recorded in children in Cameroon are caused by viruses. The present study is also the first report on the genetic variability of the G gene of HRSV and HMPV strains in the region. Although this work partially fills gaps for some information, additional studies are required to clarify the molecular epidemiology and evolutionary pattern of respiratory viruses in sub-Saharan Africa in general and more particularly in Cameroon.

Introducción: Las infecciones agudas del tracto respiratorio (IRA) son muy prevalentes, las IRAs bajas constituyen la cuarta causa de muerte a nivel mundial. Los agentes causales más comunes en niños son el Rinovirus (RV) y el Virus sincitial respiratorio (VSR). Existen tres especies de RV (A, B, C), estudios recientes han demostrado que la sintomatología y severidad de la enfermedad varía dependiendo de la especie de RV por la que hayan sido infectados los pacientes. Objetivo: Evaluar la prevalencia de Rinovirus en muestras de hisopado nasofaríngeo de niños con diagnóstico clínico de IRA en Lima, Perú durante el periodo 2009-2010. Materiales y métodos: Estudio retrospectivo de muestras de hisopado nasofaríngeo en niños, procesadas por la técnica RT-PCR para identificación de RV y sus especies. La población está compuesta por pacientes pediátricos en el Hospital Nacional Cayetano Heredia. Se analizó las variables por medio de la prueba de Chi cuadrado y Fischer. Resultados: RVA se detectó en 10.26%, RVB en 16.67%, RVC 73.9%. Grupo etario más prevalente fue de 0-5 meses. Signos y síntomas más comunes fueron tos, fiebre, rinorrea y dificultad respiratoria. Se encontró asociación entre sibilancias y RVA; tos, sibilancias e inyección conjuntival con RVC. Se halló pico de casos por RVC durante marzo, junio y noviembre. Conclusión: Se encontró alta prevalencia de infección por RVC en pacientes pediátricos, principalmente en pacientes de 0-5 meses. Distribución mensual muestra aumento de casos en marzo y junio. Se sugiere realizar vigilancia epidemiológica y estudios longitudinales para el estudio de este patógeno. / Introduction: Acute respiratory tract infections (ARTI) are a very prevalent group of diseases, lower ARTI represent the fourth cause of death worldwide. In children, the two most usual agents are Rhinovirus (RV) and Syncytial respiratory virus (SRV). RV is responsible for most is related with lower respiratory tract infections. Scientists have identified three RV species (A, B, C), recent studies have reported that symptomatology and severity vary within RV species. Objective: Asses the prevalence of Rhinovirus on nasopharyngeal swab samples of children with clinical diagnosis of ARTI in Lima, Peru during 2009-2010. Materials and method: Retrospective study about nasopharyngeal swab on children, which were processed through RT-PCR technique to identify RV and its species. The study population was pediatric patients, with clinical diagnosis of ARTI, at Hospital Nacional Cayetano Heredia. This investigation project will be revised by the ethics committee from Universidad Peruana de Ciencias Aplicadas. Results: RVA was detected in 10.26% of cases, RVB 16-67% and RVC in 73.9%. The most prevalent age group was the 0-5 months old. The most common signs and symptoms were cough, fever, rhinorrhea, and respiratory distress. The study found association between wheezing and RVA infection, cough, wheezing and conjunctival injection and RVC infection. There was a peak in RVC cases during the March, June, and November. Conclusion: We found a high prevalence for RVC infection, mainly in children between 0-5 months old. Monthly distribution showed an increase of RVC cases during March and June; epidemiological surveillance and longitudinal studies should be encouraged. / Tesis

Objetivo: Describir la distribución de patógenos responsables de IRA durante el período 2009-2010 en Cajamarca, Perú: en el contexto de la pandemia del virus de la Influenza AH1N1 2009. Resultados: Se encontraron 17 patógenos respiratorios (14 virus y 3 bacterias) en 124 niños, de los cuales el 50% eran hombres y el 50% mujeres, con un 55,6% (69/124) de 1 a 5 años y un 15,3% (19/124) de 6 a 11 años. Los patógenos más comunes fueron: Influenza A (20,2%, 25/124), Mycoplasma pneumoniae (14,5%, 18/124), RSV-A (12,1%, 15/124), Chlamydia pneumoniae (9,7%, 12/124). No se pudo identificar un patógeno específico en el 33,8% de los pacientes (42/124). Los signos y síntomas más comunes fueron fiebre 90,3%, tos 85,5%, rinorrea 74,2% y malestar 66,9%. El distrés respiratorio y la hospitalización fueron las complicaciones más comunes con 37,1% y 28,2% de los casos. Se informó una sola muerte de un niño diagnosticado con el virus de la influenza A. Además, se identificaron 12 casos de coinfecciones: 9 con 2 patógenos, 2 con tres patógenos y 1 con 4. Conclusión: Los patógenos respiratorios más comunes encontrados durante la pandemia de la Influenza AH1N1 2009-2010 fueron Influenza A, Mycoplasma pneumoniae, RSV-A y Chlamydia pneumoniae. La mayoría de patógenos presentan un perfil clínico similar Se detectaron varios casos de coinfecciones, con hasta 4 patógenos en un solo paciente. / Objective: To describe the distribution of pathogens responsible for ARI during the 2009-2010 period in Cajamarca, Peru: in the context of the 2009 AH1N1 influenza virus pandemic. Results: 17 respiratory pathogens (14 viruses and 3 bacteria) were found in 124 children, of which 50% were men and 50% women, with 55.6% (69/124) aged 1 to 5 years and a 15.3% (19/124) from 6 to 11 years old. The most common pathogens were Influenza A (20.2%, 25/124), Mycoplasma pneumoniae (14.5%, 18/124), RSV-A (12.1%, 15/124), Chlamydia pneumoniae (9,7%, 12/124). A specific pathogen could not be identified in 33.8% of patients (42/124). The most common signs and symptoms were fever 90.3%, cough 85.5%, rhinorrhea 74.2% and discomfort 66.9%. Respiratory distress and hospitalization were the most common complications with 37.1% and 28.2% of the cases. A single death was reported in a child diagnosed with influenza A virus. In addition, 12 cases of coinfections were identified: 9 with 2 pathogens, 2 with 3 pathogens, and 1 with 4. Conclusion: The most common respiratory pathogens encountered during the 2009-2010 AH1N1 influenza pandemic were influenza A, Mycoplasma pneumoniae, RSV-A, and Chlamydia pneumoniae. Most pathogens have a similar clinical profile. Several cases of coinfections were detected, with up to 4 pathogens in a single patient. / Tesis

Thesis (MScMedSc)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Background: Sudden infant death syndrome (SIDS) is considered the second most frequent cause of infant mortality worldwide. Research specifically pertaining to SIDS is limited in the South African setting. Identifiable causes for sudden infant death remain challenging despite full medico-legal investigations inclusive of autopsy, scene visit and ancillary studies. Viral infections could contribute to some sudden unexpected death in infancy (SUDI) cases, especially since a multitude of respiratory viruses have been detected from autopsy specimens. The specific contribution of viruses in the events preceding death, including the subsequent involvement of the immature immune response in infants, still warrants deciphering. Infancy is characterised by marked vulnerability to infections due to immaturities of their immune systems that may only resolve as infants grow older when these sudden deaths rarely still occur. In South Africa there is a lack of a standard protocol for investigations into the causes of SIDS, including the lack of standard guidelines as to which specimens should be taken, which viruses should be investigated and which laboratory assays should be utilised. Objectives: In this prospective descriptive study we aimed to investigate the prevalence of viruses in SUDI and SIDS cases at Tygerberg Forensic Pathology Service (FPS) Mortuary over a one year period. The primary aim was to explore possible respiratory viral infections in SUDI and SIDS cases and to determine the usefulness of molecular techniques to detect viruses from SUDI cases. To determine the significance of viruses, we assessed signs of infection from lung histology. The secondary objectives included collecting demographic data to investigate possible risk factors for SUDI and to look for possible similarities between viruses confirmed in living hospitalised infants at Tygerberg, during the study period compared to viruses detected from SUDI cases. Methods: Between May 2012 and May 2013 samples were collected from 148 SUDI cases presenting at Tygerberg FPS Mortuary. As part of the mandatory routine investigations into SUDI, shell vial culture (SVC) results were collected from lung and liver tissue specimens and bacterial culture results were collected from left and right lung and heart swabs at autopsy. To investigate the possibility of viruses implicated in some of the infant deaths we used the Seeplex® RV15 Ace detection multiplex polymerase chain reaction (PCR) assay to establish the frequency of 13 ribonucleic acid (RNA) respiratory viruses (influenza A and B, human parainfluenza 1-4, human coronavirus [OC43, 229E/NL63], human rhinovirus A, B and C, respiratory syncytial virus A and B, human enterovirus and human metapneumovirus) from RNA extracted from tracheal and lower left and right lung lobe swabs. Tissue from the lower left and right lung lobes were also assessed for histology signs of infection. Results: During our study we confirmed multiple known demographic risk factors for SIDS, such as the age peak around 1-3 months, the male predominance, bed-sharing, sleeping in the prone position, heavy wrapping in warm blankets, prenatal smoke exposure, and socio-economic factors. With the Seeplex® RV15 Ace detection assay between one and three viruses were detected in 59.5% (88/148) of cases. Of the 88 cases that had viruses detected, 75% (66/88) had one virus and 25% (22/88) had co-detections of two to three viruses. The most common viruses detected were HRV in 77% (68/88) of cases, RSV in 18% (16/88) of cases and HCoV in 14% (12/88) of cases. Many of the viruses we detected from our cases are included in the SVC test that forms part of the medico-legal laboratory investigation for all SUDI cases at Tygerberg FPS Mortuary. SVCs were positive in 9.5% (14/148) of all cases only. We showed that the SVC method is potentially missing most of the 13 respiratory viruses we investigated that could contribute to death in some of the SUDI cases. Conclusion: In some cases that had a Cause of Death Classification - SIDS, the PCR viruses detected cannot be ignored, especially when it is supported by histological evidence of infection. We thus propose that the use of PCR could alter a Cause of Death Classification from SIDS to Infection in some of these cases. Further research is needed to determine the significance of detecting viruses from SUDI cases wherein no significant histological evidence of infection was observed. This questions whether PCR may be too sensitive and is detecting past and latent viral infections that do not play any role in the cause of death. The histological picture also requires further characterisation to determine if it accurately predicts infections or lethal events and can truly support virology findings, especially in young infants whose immune systems are still maturing. Without determining the true prevalence of viruses in SUDI cases and the viral-specific immune response, the contribution of virus-specific infections to this syndrome will remain largely undetermined. / AFRIKAANSE OPSOMMING: Agtergrond: Wiegiedood (“SIDS/SUDI”) word beskou as die tweede mees algemene oorsaak van sterftes in kinders jonger as een jaar wêreldwyd. Toegewyde SIDS-spesifieke navorsing in die Suid-Afrikaanse samelewing is beperk. Dit bly steeds „n uitdaging om oorsake te probeer identifiseer vir hierdie onverwagte sterftes in kinders (SUDI) ten spyte van volledige medies-geregtelike ondersoeke, insluitende die lykskouing, ondersoek van die doodstoneel en aanvullende ondersoeke. Virusinfeksies kan aansienlik bydra tot sommige onverwagte sterftes in kinders, aangesien verskeie respiratoriese virusse alreeds aangetoon is in monsters verkry tydens outopsies. Die spesifieke rol wat virusse speel in die prosesse wat die dood voorafgaan, asook die bydraende rol van „n onder-ontwikkelde immuunrespons in babas, regverdig verdere ondersoek. Die eerste jaar van lewe word gekenmerk deur verhoogde vatbaarheid vir infeksies weens die ontwikkelende immuunstelsels soos wat babas ouer word, en die voorkoms van SUDI neem stelselmatig af met „n toename in ouderdom. In Suid-Afrika bestaan daar tans geen standaard protokol vir die ondersoek van wiegiedood nie en daar is ook nie standaard riglyne oor die tipe monsters wat geneem moet word, watter virusse ondersoek moet word en watter laboratorium toetse uitgevoer moet word nie. Doelstellings: In hierdie prospektiewe beskrywende studie is gepoog om die virusse wat in gevalle van wiegiedood of SUDI voorkom te ondersoek. Die studie is uitgevoer by die Tygerberg Geregtelike Patologie Dienste lykshuis oor 'n tydperk van een jaar. Molekulêre tegnieke om virusse aan te toon in hierdie gevalle is gebruik om spesifieke virusinfeksies te ondersoek. Die resultate is met histologiese tekens van infeksie in longweefsel gekorreleer. Demografiese data is verder versamel om moontlike risikofaktore vir wiegiedood te ondersoek. Dit is verder vergelyk met virusse wat met dieselfde diagnostiese tegnieke in babas geïdentifiseer is wat tydens die studieperiode in Tygerberg Hospitaal opgeneem was met lugweginfeksies. Metodes: Monsters van 148 SUDI gevalle wat by die Tygerberg lykshuis opgeneem is, is versamel tussen Mei 2012 en Mei 2013. As deel van die roetine ondersoeke in SUDI gevalle, was selkultuur resultate verkry van long en lewer weefsel, asook bakteriële kulture van deppers wat van beide longe en hart geneem was tydens die lykskouings. „n Seeplex® RV15 Ace polimerase kettingreaksie (PKR) toets is gebruik om die teenwoordigheid van virusse te ondersoek wat moontlik by die babasterftes betrokke kon wees. Trageale- en longdeppers wat tydens die lykskouings versamel was, was getoets vir 13 ribonukleïensure (RNS) respiratoriese virusse (influenza A and B, human parainfluenza 1-4, human coronavirus [OC43, 229E/NL63], human rhinovirus A, B and C, respiratory syncytial virus A and B, human enterovirus and human metapneumovirus). Resultate: Ons studie het verskeie bekende demografiese risikofaktore vir SUDI bevestig, byvoorbeeld „n ouderdomspiek tussen een en drie maande ouderdom, manlike predominansie, deel van „n bed met ander persone, slaap posisie op die maag, styf toedraai in warm komberse, blootstelling aan sigaretrook voor geboorte en sosio-ekonomiese faktore. Die Seeplex® RV15 Ace toets het tussen een en drie virusse geïdentifiseer in 59.5% (88/148) van die gevalle. Uit die 88 gevalle waarin virusse opgespoor was, was selgs een virus in 75% (66/88) van gevalle gevind en twee en drie virusse in 25% (22/88). Die mees algemene virusse was HRV in 77% (68/88) van gevalle, RSV in 18% (16/88) van gevalle en HCoV in 14% (12/88) van gevalle. Baie van die virusse wat tydens hierdie studie ondersoek was, was ingesluit in die roetine selkultuur toets wat deel vorm van die standaard medies-geregtelike laboratoriumondersoeke in alle SUDI gevalle by die Tygerberg lykshuis, alhoewel die selkulture positief was in slegs 9.5% (14/148) van gevalle. Ons het gevind dat baie respiratoriese virusse potensieel gemisdiagnoseer word wat „n rol kon speel in of bydra tot die dood van sommige SUDI gevalle. Gevolgtrekking: In sommige gevalle waarin SIDS geklassifiseer is as die oorsaak van dood, kan die virusse wat met PKR toetse opgespoor is nie geïgnoreer word nie, veral waar die bevinding ondersteun word deur histologiese bewyse van infeksie. Ons stel dus voor dat die gebruik van PKR toetse die oorsaak van dood klassifikasie kan verander van SIDS na Infeksie in sommige van hierdie gevalle. Verdere navorsing is nodig om die waarde van gelyktydige opsporing van virusse in SUDI gevalle te bepaal wanneer daar geen noemenswaardige histologiese bewyse van infeksie gevind word nie. Dit bevraagteken of die PKR toets dalk te sensitief is en gevolglik vorige en latente virusinfeksies identifiseer wat nie noodwendig 'n rol in die oorsaak van dood speel nie. Die diagnostiese en kliniese waarde van die histologiese beeld in terme van die rol van virusinfeksies as bydraende oorsaak van dood moet verder ondersoek word, veral in jong kinders wie se immuunstelsels nog nie volledig ontwikkel is nie. Indien die werklike voorkoms van virusse in SUDI gevalle en die virus-spesifieke immuunrespons nie bepaal word nie, sal die rol van virus-spesifieke infeksies in hierdie sindroom grootliks onbekend bly. / Harry Crossley Foundation / Poliomyelitis Research Foundation (PRF) / National Health Laboratory Services Research Trust

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The most common viruses involved in acute respiratory diseases among young children are the respiratory syncytial virus (RSV), influenzavirus (FLU), parainfluenzavirus (PIV), adenovirus (AdV) and human rhinovirus (HRV). The purpose of the present study was to identify the main respiratory viruses that affected children younger than five years old in Uberlândia, in Midwestern Brazil. Nasopharyngeal aspirates from 379 children attended at Hospital de Clínicas (HC/UFU), from 2001 to 2004, with acute respiratory disease, were collected and tested by either immunofluorescence assay (IFA) or reverse transcription polymerase chain reaction (RT-PCR). RSV was detected in 26.4% (100/379) of samples, FLU A and B in 9.5% (36/379), PIV 1, 2 and 3 in 6.3% (24/379) and AdV in 3.7% (14/379). Negative and indeterminate samples (205) by IFA were tested by RT-PCR for detection of HRV, and 29.6% (112/379) was positive. RSV, particularly among children in their first 6 months of life, and HRV cases showed highest incidence. For both virus, bronchiolitis and pneumonia/bronchopneumonia were the main nosological presentation among children less than 6 months, were RSV responded for 40,3% and 34,6% of the cases, respectively, and the HRV were detected in 25,0% of bronchiolitis cases and 15,4% of pneumonia/bronchopneumonia cases at the same referred age group. Negative samples by both IFA and RT-PCR might be indicative that other pathogens, such as coronavirus, human metapneumovirus and bacteria, could be the causative agent in these infections. Laboratorial diagnosis constituted an essential instrument to determine the incidence of the most common viruses in respiratory infections among young children in this region. / Os vírus mais comumente envolvidos em doenças respiratórias agudas em crianças são os vírus respiratório sincicial (VRS), influenzavírus tipos A e B (FLU A e B), parainfluenzavírus tipos 1, 2 e 3 (PIV-1, -2 e -3), adenovírus (AdV) e os rinovírus (HRV). O objetivo geral do presente estudo foi identificar os principais vírus respiratórios envolvidos em doenças respiratórias aguda (DRA) em crianças de até cinco anos de idade no período de 2001 a 2004. Aspirados de nasofaringe de 379 crianças atendidas no Hospital de Clínicas da Universidade Federal de Uberlândia (HC/UFU) com doença respiratória aguda foram coletadas e testadas pelas reações de imunofluorescência indireta (IFI) ou transcrição reversa da reação em cadeia da polimerase (RT-PCR). O VRS foi detectado em 26,4% (100/379) dessas amostras, FLU tipos A e B em 9,5% (36/379), PIV 1, 2 e 3 em 6,3% (24/379) e AdV em 3,7% (14/379). As amostras negativas e inconclusivas pela IFI (205) foram testadas por RT-PCR para detecção dos HRV, sendo que 26,9% (112/379) foram positivas. Neste estudo, os vírus mais comumente detectados em quadros clínicos de bronquiolite e pneumonia/broncopneumonia em crianças menores de seis meses de idade foram o VRS, respondendo por 40,3% e 34,6% do total de casos, respectivamente, e o HRV, identificado em 25,0% dos casos de bronquiolite e em 15,4% dos casos de pneumonia/broncopneumonia na mesma faixa etária. Relacionado à sazonalidade, a circulação dos vírus identificados predominou nos meses de temperaturas mais baixas. Esse padrão foi evidente para o VRS, que apresentou um pico de ocorrência em abril e maio, enquanto que o pico do FLU e HRV ocorreu em julho. Amostras negativas tanto pela IFI quanto pela RT-PCR indicam que outros patógenos, incluindo coronavírus, metapneumovirus humano além de bactérias, podem ter sido os responsáveis pelas infecções. Finalizando, este estudo foi essencial para um diagnóstico conclusivo de DRAs causadas por vírus, bem como para determinar quais agentes virais circularam nesta região. / Mestre em Imunologia e Parasitologia Aplicadas

In Brazil, the few studies conducted have used viral etiology, in general, traditional methods (imunofluorescense techniques and and viral cultures) in hospitalized children. The purpose of the present study was to investigate the presence of respiratory viruses using indirect immunofluorescence techniques and the reverse transcription followed by polymerase chain reaction in nasopharyngeal aspirates of children with acute respiratory disease attendet in public institutions of primary and secondary care in the city of Uberlândia. Between february, 2008 to may, 2010 were obtained a convenience sample, nasopharyngeal aspirates from children under five years old with symptoms of acute respiratory disease, attended at Unidade Básica de Saúde da Família - Granada 1, Unidade de Atendimento Integrado-Pampulha and in Clinica Infantil Don Bosco in Uberlândia. Acute respiratory disease was defined by the presence of coryza, coughing, breathing difficulties and/or sibilance, with or without fever. The indirect immunofluorescence techniques and the reverse transcription followed by polymerase chain reaction were used to test for the presence of respiratory viruses. A total of 43 children (53,5% male and 46,5% female) between two and 60 months of age (average: 18,3 months; median 15 months; DP±16). The clinical diagnosis for admission was common cold for 23 children (53,4%), tracheobronchitis in four (9,3%); pneumonia in 12 (28%) and bronchiolitis in four (9,3%). At least one respiratory virus was detected in 22 (51,1%) of the samples. A total of 26 viruses were identified. Ten (38,4%) samples were positive for the respiratory syncytial virus; ten (38,4%) for rhinovirus, three (11,5%) for parainfluenzavirus; two (7,7%) for adenovirus and one (3,8%) for influenzavirus. Co-infection occurred in three of the samples. The indirect immunofluorescence techniques identified nine (21,0%) and the reverse transcription followed by polymerase chain reaction 19 (44,1%) of the respiratory viruses. The rhinovirus and respiratory syncytial virus were the respiratory virus most prevalent in children with acute respiratory disease in public institutions of primary and secondary care. The use of molecular method permitted a two fold increase in the capacity for detection of the viral agent collected from the nasopharyngeal aspirates. / No Brasil, os poucos estudos conduzidos com o objetivo de verificar a etiologia viral das doenças respiratórias agudas utilizaram, em geral, métodos tradicionais (imunofluorescência e cultura viral) em crianças hospitalizadas. O objetivo geral do presente estudo foi investigar a presença de vírus respiratórios por meio das técnicas de imunofluorescência indireta e da transcrição reversa seguida da reação em cadeia da polimerase em aspirados de nasofaringe de crianças com doença respiratória aguda atendidas em serviços públicos de atenção primária e secundária na cidade de Uberlândia, MG. Entre fevereiro de 2008 a maio de 2010 foram obtidos, por meio de uma amostra de conveniência, aspirados de nasofaringe de crianças menores de cinco anos com sintomas de doença respiratória aguda, atendidas na Unidade Básica de Saúde da Família - Granada 1, Unidade de Atendimento Integrado-Pampulha e Clínica Infantil Dom Bosco Uberlândia. Doença respiratória aguda foi definida pela presença de coriza, tosse, dificuldade para respirar ou sibilância, com ou sem febre. A imunofluorescência e a transcrição reversa seguida da reação em cadeia da polimerase foram utilizadas para testar a presença de vírus respiratórios. Partiparam do estudo 43 crianças (53,5% do gênero masculino e 46,5% gênero feminino) com idade entre dois a 60 meses (média = 18,3 meses; mediana = 15 meses; DP=±16). O diagnóstico clínico à admissão foi de resfriado comum em 23 crianças (53,4%), traqueobronquite, em quatro (9,3%), pneumonia, em doze (28%) e bronquiolite, em quatro (9,3%). Pelo menos um vírus respiratório foi detectado em 22 (51,1%) amostras, sendo que 26 vírus foram identificados. Dez (38,4%) amostras foram positivas para o vírus respiratório sincicial, dez (38,4%) para o rinivirus, três (11,5%) para o parainfluenzavírus; duas (7,7%) para adenovírus e uma (3,8%), para o influenzavírus. A presença de co-infecção ocorreu em três amostras. A imunofluorescência identificou nove (21%) e a transcrição reversa seguida da reação em cadeia da polimerase dezenove (44,1%) vírus respiratórios. O rinovírus e o vírus respiratório sincicial foram os vírus mais prevalentes em crianças com doença respiratória aguda em serviços de atenção primária e secundária. A utilização de método molecular permitiu dobrar a capacidade de detecção de agente viral nos aspirados de nasofaringe. / Mestre em Ciências da Saúde

INTRODUÇÃO: A adesão à vacinação contra influenza é historicamente baixa entre profissionais da área da saúde (PAS) (2 a 36%). A ocorrência de sintomas respiratórios após vacinação é freqüentemente interpretada como falha vacinal. No Hospital das Clínicas da FMUSP, um estudo preliminar mostrou que as principais razões para não adesão são a percepção da ineficácia da vacina e o medo de reações adversas. OBJETIVOS: Identificar a incidência de eventos adversos pós-vacinação e identificar os vírus respiratórios (VR) responsáveis por eventuais episódios de infecção de via aérea superior (IVAS) que ocorram entre indivíduos vacinados. MÉTODOS: Foi seguida uma coorte de 398 PAS vacinados objetivando verificar a ocorrência de eventos adversos até 48 h após a vacinação. Durante 4 meses, 337 PAS foram seguidos 2 vezes por semana para avaliar a ocorrência de sintomas respiratórios. Lavados nasais foram coletados na presença de sintomas para pesquisa de VR. A técnica de imunofluorescência direta foi usada para diagnosticar vírus sincicial respiratório, influenza A e B, adenovírus e parainfluenza. PCR foi utilizada para detectar picornavírus e coronavírus e PCR em tempo real para diagnosticar metapneumovírus. Para assegurar melhor sensibilidade, influenza A e B foi também detectado pela PCR em tempo real e adenovírus pela PCR. RESULTADOS: Eventos adversos foram relatados por 30% dos PAS, predominando cefaléia (15,1%), mialgia (14,3%) e mal estar (13,6%). Nenhum evento adverso grave foi observado. Cento e vinte e um PAS (35,9%) desenvolveram sintoma respiratório durante o seguimento e lavado nasal foi colhido em 93 dos 192 episódios apresentados. Vírus influenza A foi detectado em 5 dos 93 episódios (5,3%) e outros vírus respiratórios em 26 (27,9%). No restante dos 61 episódios (65,6%) nenhum vírus foi encontrado. A densidade de incidência de infecção pelo vírus influenza foi de 4,3 episódios por 100 pacientes-mês enquanto que a densidade de infecção por outros vírus respiratórios foi de 10,8 episódios por pacientes-mês. CONCLUSÃO: Vacina da influenza é segura. O medo de eventos adversos grave parece injustificado, bem como, a percepção da ineficácia da vacina. O presente estudo evidencia que IVAS após vacinação é predominantemente causada por outros vírus respiratórios (28%) e não pelo vírus influenza (5%) / INTRODUCTION: Compliance with influenza vaccination has been historically poor among health care workers (HCW), ranging from 2 to 36% world around. The occurrence of respiratory symptoms following influenza vaccination is frequently taken as vaccine failure which reinforces vaccine disbelief. A preliminary study conducted at Hospital das Clínicas, University of São Paulo School of Medical Sciences, showed that the main reasons for non-compliance with influenza vaccination were the perception of vaccine inefficacy and fear of adverse events. OBJECTIVES: To determine the incidence of adverse events after seasonal influenza vaccination and identify other respiratory viruses causing upper respiratory infections in vaccinated HCWs. METHODS: A cohort of 398 vaccinated HCWs was prospectively surveyed for the occurrence of any adverse event in the first 48h after vaccination. A subset of the original cohort (337 HCWs) was followed up during four months, twice a week, for the detection of respiratory symptoms. Nasal washes were taken if respiratory symptoms occurred. Direct immunofluorescent assay (DFA) was performed for the detection of respiratory syncytial virus (RSV), influenza (INF) A and B, parainfluenza (PIV) 1, 2 and 3, and adenovirus (ADV). PCR was performed for the detection of human rhinoviruses (HRV), ADV and coronaviruses (hCoV); and real time PCR for the detection of human metapneumovirus (hMPV). To assure greatest sensitivity of influenza diagnosis, real time PCR was added to the diagnostic tools of influenza viruses. RESULTS: Adverse events were reported by 30% of the HCWs, being headache and myalgia reported by 50% and 47% of the participants, respectively. No severe adverse event was observed. One hundred and twenty-one HCWs (35.9%) developed 192 episodes of respiratory symptoms during follow-up and nasal washes were taken in 93 of them. Influenza A virus was detected in five of the 93 episodes (5.3%) and other respiratory viruses in 26 (27.9%). In the remaining 61 episodes (65.6%) no respiratory virus was identified. The incidence density of influenza was 4.3 episodes per 100 HCW-month, while the incidence density of other respiratory viruses was 10.8 episodes per HCW-month. CONCLUSIONS: Influenza vaccine is safe. The fear of adverse events as well as the perception of vaccine inefficacy seems to be unjustified in this population. The present study showed that the occurrence of upper respiratory infection during the four months following seasonal influenza vaccination of HCWs is generally caused by other respiratory viruses (28%) and not by influenza viruses (5%)

INTRODUÇÃO: A adesão à vacinação contra influenza é historicamente baixa entre profissionais da área da saúde (PAS) (2 a 36%). A ocorrência de sintomas respiratórios após vacinação é freqüentemente interpretada como falha vacinal. No Hospital das Clínicas da FMUSP, um estudo preliminar mostrou que as principais razões para não adesão são a percepção da ineficácia da vacina e o medo de reações adversas. OBJETIVOS: Identificar a incidência de eventos adversos pós-vacinação e identificar os vírus respiratórios (VR) responsáveis por eventuais episódios de infecção de via aérea superior (IVAS) que ocorram entre indivíduos vacinados. MÉTODOS: Foi seguida uma coorte de 398 PAS vacinados objetivando verificar a ocorrência de eventos adversos até 48 h após a vacinação. Durante 4 meses, 337 PAS foram seguidos 2 vezes por semana para avaliar a ocorrência de sintomas respiratórios. Lavados nasais foram coletados na presença de sintomas para pesquisa de VR. A técnica de imunofluorescência direta foi usada para diagnosticar vírus sincicial respiratório, influenza A e B, adenovírus e parainfluenza. PCR foi utilizada para detectar picornavírus e coronavírus e PCR em tempo real para diagnosticar metapneumovírus. Para assegurar melhor sensibilidade, influenza A e B foi também detectado pela PCR em tempo real e adenovírus pela PCR. RESULTADOS: Eventos adversos foram relatados por 30% dos PAS, predominando cefaléia (15,1%), mialgia (14,3%) e mal estar (13,6%). Nenhum evento adverso grave foi observado. Cento e vinte e um PAS (35,9%) desenvolveram sintoma respiratório durante o seguimento e lavado nasal foi colhido em 93 dos 192 episódios apresentados. Vírus influenza A foi detectado em 5 dos 93 episódios (5,3%) e outros vírus respiratórios em 26 (27,9%). No restante dos 61 episódios (65,6%) nenhum vírus foi encontrado. A densidade de incidência de infecção pelo vírus influenza foi de 4,3 episódios por 100 pacientes-mês enquanto que a densidade de infecção por outros vírus respiratórios foi de 10,8 episódios por pacientes-mês. CONCLUSÃO: Vacina da influenza é segura. O medo de eventos adversos grave parece injustificado, bem como, a percepção da ineficácia da vacina. O presente estudo evidencia que IVAS após vacinação é predominantemente causada por outros vírus respiratórios (28%) e não pelo vírus influenza (5%) / INTRODUCTION: Compliance with influenza vaccination has been historically poor among health care workers (HCW), ranging from 2 to 36% world around. The occurrence of respiratory symptoms following influenza vaccination is frequently taken as vaccine failure which reinforces vaccine disbelief. A preliminary study conducted at Hospital das Clínicas, University of São Paulo School of Medical Sciences, showed that the main reasons for non-compliance with influenza vaccination were the perception of vaccine inefficacy and fear of adverse events. OBJECTIVES: To determine the incidence of adverse events after seasonal influenza vaccination and identify other respiratory viruses causing upper respiratory infections in vaccinated HCWs. METHODS: A cohort of 398 vaccinated HCWs was prospectively surveyed for the occurrence of any adverse event in the first 48h after vaccination. A subset of the original cohort (337 HCWs) was followed up during four months, twice a week, for the detection of respiratory symptoms. Nasal washes were taken if respiratory symptoms occurred. Direct immunofluorescent assay (DFA) was performed for the detection of respiratory syncytial virus (RSV), influenza (INF) A and B, parainfluenza (PIV) 1, 2 and 3, and adenovirus (ADV). PCR was performed for the detection of human rhinoviruses (HRV), ADV and coronaviruses (hCoV); and real time PCR for the detection of human metapneumovirus (hMPV). To assure greatest sensitivity of influenza diagnosis, real time PCR was added to the diagnostic tools of influenza viruses. RESULTS: Adverse events were reported by 30% of the HCWs, being headache and myalgia reported by 50% and 47% of the participants, respectively. No severe adverse event was observed. One hundred and twenty-one HCWs (35.9%) developed 192 episodes of respiratory symptoms during follow-up and nasal washes were taken in 93 of them. Influenza A virus was detected in five of the 93 episodes (5.3%) and other respiratory viruses in 26 (27.9%). In the remaining 61 episodes (65.6%) no respiratory virus was identified. The incidence density of influenza was 4.3 episodes per 100 HCW-month, while the incidence density of other respiratory viruses was 10.8 episodes per HCW-month. CONCLUSIONS: Influenza vaccine is safe. The fear of adverse events as well as the perception of vaccine inefficacy seems to be unjustified in this population. The present study showed that the occurrence of upper respiratory infection during the four months following seasonal influenza vaccination of HCWs is generally caused by other respiratory viruses (28%) and not by influenza viruses (5%)