<p>Uvod: Virusni gastrointestinalni sindrom je aktuelni zdravstveni problem u celom svetu. To važi kako u razvijenim zemljama, tako i u zemljama u razvoju, a posebno u nerazvijenim zemljama, gde je drugi po redu uzrok mortaliteta. Nagli početak bolesti, praćen pojavom velikog broja tečnih stolica, mukom, povraćanjem, bolovima u stomaku, temperaturom, malaksalo&scaron;ću, ima za posledicu dehidrataciju. U svim starosnim grupama obolelih, a naročito kod sasvim male dece, starih i imunodeficitarnih osoba može da dođe do smrtnog ishoda, ukoliko se brzo ne postavi tačna etiolo&scaron;ka dijagnoza bolesti i ne pristupi se odmah nadoknadi vode i elektrolita, kao i primeni svih ostalih mera simptomatske terapije. Brzo postavljena tačna dijagnoza, &scaron;to se najbolje postiže real-time PCR testom, sprečava pojavu komplikacija, pa i fatalnog ishoda bolesti. Istovremeno, omogućava primenu odgovarajućih epidemiolo&scaron;kih mera da se spreči nastanak epidemija i njihovo &scaron;irenje. Cilj ovog istraživanja bio je da se tačno utvrdi incidenca virusnog gastrointestinalnog sindroma u Vojvodini i učestalost pojave epidemijskog i sporadičnog javljanja ove bolesti. Cilj je bio i postavljanje algoritma za primenu real-time PCR testa u dijagnostici virusnog gastrointestinalnog sindroma u budućem radu. Isto tako, cilj je bio da se molekularnom analizom, sekvenciranjem delova genoma pozitivnih uzoraka stolice, izvr&scaron;i genetska tipizacija i odredi filogenetska pripadnost virusa. Materijal i metode: Tokom petogodi&scaron;njeg istraživanja molekularnim real-time PCR testom pregledane su 1003 obolele osobe sa simptomima virusnog dijarealnog sindroma, starosti od mesec dana do preko 90 godina. Pregledani su na rota, noro, astro i enterične adenoviruse. Na osnovu podataka iz anketnih upitnika i istorija bolesti, detaljno su analizirani svi klinički pokazatelji (javljanje bolesti tokom godine, trajanje bolesti, simptomi). Procena težine kliničke slike vr&scaron;ena je prema Vesikari skali. Svi podaci su upoređivani prema vrsti virusnog uzročnika, prema starosti obolelih, godinama trajanja istraživanja i epidemijskom i sporadičnom javljanju oboljenja. Dobijeni podaci su statistički obrađeni, tabelarno i grafički prikazani. Rezultati: U petogodi&scaron;njem periodu real-time PCR testom pregledan je uzorak od 1003 obolele osobe različite starosti na 4 virusna uzročnika dijarealnog sindroma (rota, noro, astro i enterične adenoviruse). Virusni dijarealni sindrom dokazan je kod 709 obolelih (70,69%). Najče&scaron;će su dokazane rotavirusne infekcije u 28,81%. Statistički značajno najče&scaron;će rotavirusi su bili utvrđeni kod dece do 5 godina (38,90%), ali u visokom procentu i kod dece uzrasta 6 do 14 godina (24,83%). Deca mlađa od 5 godina imala su statistički značajno najtežu kliničku sliku, bila su če&scaron;će hospitalizovana i imala su statistički značajno vi&scaron;u temperaturu. Pored vi&scaron;e temperature kod obolelih od rotavirusa, klinička slika je kod ovih bolesnika bila teža i bolest je duže trajala nego kod obolelih od drugih virusa. Norovirusna infekcija je dokazana u 23,03% obolelih i to statistički značajno če&scaron;će kod odraslih osoba, starijih od 20 godina. Od kliničkih simptoma kod ovih bolesnika statistički značajno če&scaron;će su dokazani muka, povraćanje i bolovi u stomaku, nego kod obolelih od drugih virusa. Norovirusi su značajno če&scaron;će bili uzročnici epidemijskog javljanja bolesti. Astrovirus je dokazan kod znatno manjeg broja obolelih (u 2,29%) i to samo kod dece do 5 godina i dece uzrasta 6 do 14 godina. Infekcija izazvana enteričnim adenovirusima dokazana je kod 13,36% bolesnika. Njače&scaron;će je utvrđena kod dece uzrsta do 5 godina i 6 do 14 godina. Oboleli od adenovirusa imali su statistički značajno blažu kliničku sliku bolesti. Dva virusna uzročnika u uzorku stolice dokazana su u 3,19% osoba, obično u toku epidemijskog javljanja bolesti. Ovi bolesnici su imali bitno težu kliničku sliku. Najvi&scaron;e obolelih od dijarealnog sindroma bilo je u hladnim mesecima, mada su dijagnostikovani i tokom cele godine. U petogodi&scaron;njem periodu utvrđene su 22 epidemije u kolektivima i 9 porodičnih epidemija. Epidemijsko javljanje bolesti bilo je statistički značajno najče&scaron;će kod najstarijih bolesnika (starijih od 50 godina), a sporadično javljanje bilo je statističko značajno najče&scaron;će kod dece. U cilju potvrde tačnosti dijagnostike virusa u ispitivanim uzorcima real-time PCR testom, genotipizacije, kao i detaljnije molekularne analize, izabrani su reprezentativni uzorci pozitivni na rota, noro, astro ili adenoviruse. Delovi genoma ovih uzoraka su amplifikovani, a zatim sekvencirani. Sekvencirani izolati rotavirusa pripadali su grupi A i tipovima G1P[8], G2P[4], G3P[8] i G9P[8]. Sekvencirani izolati norovirusa pripadali su genogrupi I tipu 2, zatim genogrupi II tipovima 1, 2, 4 i 17. Sekvencirani izolati astrovirusa pripadali su grupi klasičnih astrovirusa i tipovima 1, 4 i 5. Sekvencirani izolati adenovirusa pripadali su grupi F i tipovima 40 i 41, kao i grupi C tipu 2. Pripadnost dobijenih sekvenci u ovom istraživanju, dodatno je potvrđena izradom filogenetskog stabla za sekvence pozitivne na rota, noro, astro ili adenoviruse. Zaključak: Incidenca virusnog dijarealnog sindroma u Vojvodini (70,69%) vrlo je visoka i vi&scaron;a je nego &scaron;to je bilo pretpostavljeno prilikom prijave teze (u hipotezi). Real-time PCR test treba da bude redovno kori&scaron;ćen u budućem dijagnostičkom radu, jer dovodi do brze dijagnostike, čak i ako su virusi prisutni u malom broju u uzorcima tečnih stolica, &scaron;to je utvrđeno tokom ovog dijagnostičkog rada. Ispitivani virusi treba da budu redovno dijagnostikovani kod obolelih od dijarealnog sindroma i to u svim starosnim grupama, tokom epidemijskog i sporadičnog javljanja oboljenja.</p> / <p>Introduction: Viral gastrointestinal syndrome is a current ongoing health problem worldwide. This is true of both developed and developing countries, especially underdeveloped ones where it is the second leading cause of mortality. Sudden onset of the disease&mdash;accompanied by the occurrence of large numbers of liquid stools, nausea, vomiting, abdominal pain, fever, and exhaustion&mdash;leads to dehydration. A fatal outcome can occur in all age groups of patients, especially very young children, the elderly, and the immuno-deficient, unless an accurate etiological diagnosis of the disease is quickly established, followed by a prompt institution of fluid and electrolyte placement, and implementation of other symptomatic therapy measures. Quick establishment of an accurate diagnosis, which is best achieved using the real-time PCR test, prevents the onset of complications, including a potentially fatal outcome of the disease. Simultaneously, it enables the implementation of appropriate epidemiological measures to prevent epidemic outbreaks and their spread. The aim of this study was to accurately determine the incidence of viral gastrointestinal syndrome in Vojvodina and the frequency of epidemic and sporadic occurrence of this disease. The aim was also to set up an algorithm for the application of the real-time PCR test in diagnostics of viral gastrointestinal syndrome in future work. Likewise, the aim was to carry out genetic typing and determine phylogenetic affiliation of the virus using molecular analysis and sequencing of parts of genomes from positive stool samples. Material and Methods: During a five-year study, 1003 patients with symptoms of viral diarrheal syndrome, aged from one month to more than 90 years old, were examined using molecular real-time PCR test. They were screened for rota, noro, astro, and enteric adenoviruses. Based on the data from survey questionnaires and medical case history, all clinical indicators were meticulously analyzed (disease occurrence during the year, disease duration, symptoms). The assessment of the clinical severity was carried out according to the Vesikari Clinical Severity Scoring scale. All data were compared according to the type of the viral causing agent, age of the patients, duration of research in years, and epidemic and sporadic occurrence of the disease. Obtained data were statistically analyzed, tabulated, and graphically displayed. Results: In a five-year period, a sample of 1003 patients of different ages was screened for four different viral causing agents of diarrheal syndrome (rota, noro, astro, and enteric adenoviruses) using the real-time PCR test. Viral diarrheal syndrome was confirmed in 709 patients (70.69%). The most commonly found were rotavirus infections in 28.81% of the cases. Rotaviruses were statistically significantly most common in children younger than 5 years old (38.90%), but were also found in high percentage in children aged 6-14 years old (24.83%). Children under 5 years of age had statistically significantly highest clinical severity and fever, and were more frequently hospitalized. In addition to higher fever in patients with rotavirus, clinical severity in these patients was also higher, and the disease lasted longer than in patients with other viruses. Norovirus infections were reported in 23.03% of the subjects, statistically significantly more frequently in adults over 20 years of age. Regarding the clinical symptoms in these patients, nausea, vomiting, and abdominal pain were statistically significantly more common than in patients with other viruses. Noroviruses were significantly more common as causing agents of epidemic disease outbreaks. Astrovirus was found in a significantly smaller number of patients (in 2.29%), and only in children under 5 years of age and children aged 6-14 years old. Enteric adenovirus infections were reported in 13.36% of the subjects. They were most commonly found in children younger than 5, and those aged 6- 14 years old. Adenovirus sufferers had statistically significantly milder clinical disease. Two viral causing agents in the stool sample were found in 3.19% of the subjects, usually during an epidemic disease outbreak. These patients had a significantly more severe clinical disease. Highest numbers of sufferers from diarrheal syndrome occurred during the cold months, although they were diagnosed throughout the year. In a five-year period, 22epidemics in collective groups and 9 family epidemics were identified. Epidemic outbreaks of the disease were statistically significantly most frequent in the elderly patients (older than 50), while sporadic occurrences were statistically significantly most frequent in children. Representative samples positive for rota, noro, astro, or adenoviruses were selected in order to confirm the accuracy of virus diagnostics in samples tested by the real-time PCR test, and perform genotyping as well as more detailed molecular analyses. Parts of the genomes of these samples were amplified and then sequenced. Sequenced rotavirus isolates belonged to group A and types G1P[8], G2P[4], G3P[8], and G9P[8]. Sequenced norovirus isolates belonged to genogroup I type 2, and genogroup II types 1, 2, 4, and 17. Sequenced astrovirus isolates belonged to the group of classical astroviruses and types 1, 4, and 5. Sequenced adenovirus isolates belonged to group F and types 40 and 41, as well as group C type 2. The affiliation of the obtained sequences in this study was further confirmed by creating a phylogenetic tree for sequences positive for rota, noro, astro, or adenoviruses. Conclusion: The incidence of viral diarrheal syndrome in Vojvodina (70.69%) is very high&mdash;higher than what was assumed at the time of the thesis submission (in the hypothesis). The real-time PCR test should be regularly used in future diagnostic work, since it leads to rapid diagnostics even if viruses are present in small numbers in liquid stool samples, as determined in the course of this diagnostic study. The investigated viruses should be regularly tested in patients with diarrheal syndrome belonging to all age groups during both epidemic and sporadic occurrences of the disease.</p>

Propósito: Evaluar el valor del Vesikari Scoring System (VSS) como herramienta diagnóstica en la predicción de patógenos en niños con gastroenteritis aguda. Métodos: En un estudio retrospectivo, se analizó 247 historias clínicas y registros laboratoriales del Hospital de Emergencias Pediátricas (Lima, Perú), utilizando el diagnóstico de CIE-10 A00-A009 (enfermedades intestinales infecciosas). Se dividió según los patógenos detectados: no específico, viral y bacteriana. Se detectaron las bacterianas por coprocultivo y las virales por inmunoanálisis. Los casos no específicos tenían ambos resultados negativos o no hubo pruebas laboratoriales. Se ha calculado la sensibilidad, especificidad, valores predictivos negativos (VPN) y positivos (VPP), índice de probabilidad (LR) y el área bajo la curva (ROC). Resultado: El área bajo la curva ROC del VSS no fue significativo para determinar la etiología bacteriana ni viral. En el punto de corte de 9 en VSS, se analizó VPP (18.29%) y VPN (89.47%). Los leucocitos en heces tuvieron un área bajo la curva ROC de 0.8831(IC95% 0,79-0,96 y p=0.04) para etiología bacteriana, con una sensibilidad de 80.95%, especificidad de 88.24% y LR- de 0.22. Los leucocitos en heces con un punto de corte de ≥20, tuvo una precisión aceptable para diagnosticar gastroenteritis bacteriana. La prueba de moco en heces tuvo una sensibilidad de 83.33%, especificidad de 82.35% y VPP de 95.89%. Conclusión: El VSS no es una herramienta adecuada para determinar etiología de gastroenteritis viral y bacteriana. Sin embargo, se ha determinado que los leucocitos en heces con un punto de corte de ≥20 leucocitos por campo y moco en heces permite identificar la gastroenteritis aguda bacteriana. / Purpose: To evaluate the Vesikari Scoring System (VSS) as a valuable diagnostic tool for predicting pathogens in children with acute gastroenteritis. Methods: In this retrospective study were analyzed 247 clinical and laboratory records of the Pediatric Emergency Hospital (Lima, Peru), with the diagnosis of ICD-10 A00-A009 (Intestinal Infectious diseases). According to the pathogens detected were divided into groups: non-specific, viral, and bacterial. Bacteria were detected by stool culture, viral pathogens by immune analysis. The non-specific group had negative on both tests, or they did not perform. We calculated sensitivity, specificity, negative and positive predictive values, negative and positive likelihood ratios, and receiver operating characteristic curves.  Results: The area under the ROC curve of VSS was not significant for viral and bacterial etiology. The cut-off point of 9 in VSS was calculated by analyzing PPV (18,29%) and NPV (89,47%). The fecal leukocyte had a ROC of 0.8831 (IC95% 0.79-0.96 and p=0.04) for bacterial group, with a sensibility of 80,95%, specificity of 88,24% and LR- of 0,22. At a cut-off point of ≥20 fecal leukocytes was an acceptable diagnostic accuracy for bacterial gastroenteritis. The stool mucus for the bacterial group had a sensibility of 83,33%, a specificity of 82,35%, and a PPV of 95,89%. Conclusion: VSS is not an adequate diagnostic tool to differentiate viral from bacterial etiology. Nevertheless, we determine that a test of fecal leukocytes, with a cut-off point of ≥20 leukocytes per field and mucus in stools, allow us to identify the bacterial etiology of gastroenteritis. / Tesis

Thesis (MCurr)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Background Diarrhoea (frequent, loose, watery stools) is one of the major causes of morbidity and mortality globally and affects mainly infants and children under the age of five years. Unhygienic feeding practices and feeding utensils contribute to diarrhoeal incidences. The most common causes of acute gastroenteritis worldwide are infectious agents, such as viruses, bacteria and parasites Aim The aim of the study was to investigate which out of three particular sterilising methods is the most effective for sterilising feeding bottles and cups. Methods An experimental quantitative approach was most appropriate for the current study. An in vitro experimental study with a descriptive design was utilised under controlled laboratory conditions. The study was conducted at the University of the Western Cape (UWC) in April 2009. Results The sample size consisted of 16 samples, of which two were used for each method of sterilisation, namely: two (2) bottles and two (2) cups for sunlight; two (2) bottles and two (2) cups for Milton™; two (2) bottles and two (2) cups for Sunlight™ dishwashing liquid; and control utensils that consisted of two (2) bottles and two (2) cups. The target population for the study comprised infant feeding bottles and feeding cups. The analysis for the APC cultures that was compared in the cups vs. bottles, in order to see whether there was a significant difference between the mean bacteria counts, shows that the average bacteria count (on the ln scale) was 6 cfu/ml and 9 cfu/ml for the cups and bottles, respectively. The t-value was -1.17524. As the ρ-value was 0.2595, no significant difference was found between the cups and bottles. The E. coli cultures were compared in the cups vs. bottles to see whether there was a significant difference between the mean bacteria counts. The results show that the average bacteria count (on the ln scale) was 7 cfu/ml and 7.6 cfu/ml for cups and bottles, respectively. The t-value was -0.211902. The ρ-value was 0.835237, and therefore there was no significant difference between cups and bottles. Conclusion The current study showed no significant difference between the sterilising methods or between the use of either bottles or cups. Therefore, a study with a larger sample size is recommended for further research. Recommendations The researcher recommends that future researchers conduct broader studies, with a larger sample size on the topic. Studies with a larger sample size enabled the real differences to be large enough to be significant. The use of sunlight is recommended as a sterilisation method for infant feeding utensils, as it is both time- and cost-effective. Sunlight is an inexpensive and readily available method of sterilisation; therefore, it can be used by relatively under resourced socio-economic communities. / AFRIKAANSE OPSOMMING: Agtergrond Diarree (gereelde, los, waterige stoelgang) is een van die hoofoorsake van morbiditeit en sterflikheid wêreldwyd en affekteer hoofsaaklik suigelinge en kinders onder die ouderdom van vyf jaar. Onhigiëniese voedingspraktyke en -gereedskap dra by tot die voorkoms van diarree-gevalle. Die mees algemene oorsake van akute gastroënteritus wêreldwyd word veroorsaak deur aansteeklike agente soos virusse, bakterieë en parasiete. Doel Die doel van hierdie studie is om ondersoek te doen na watter van die drie bepaalde steriliseringsmetodes die mees effektiewe is vir die sterilisering van bottels en koppies. Metodes ’n Eksperimentele kwantitatiewe benadering is die mees geskikte een vir die huidige studie. ’n In vitro-eksperimentele studie met ’n deskriptiewe ontwerp is onder gekontroleerde laboratorium omstandighede aangewend. Die studie is by die Universitet van die Wes-Kaap (UWK) in April 2009 uitgevoer. Resultate Die steekproefgroote het bestaan uit 16 monsters waarvan twee gebruik is vir elke steriliseringsmetode, naamlik: twee (2) bottels en twee (2) koppies vir sonlig; twee (2) bottels en twee (2) koppies vir Milton™; twee (2) bottels en twee (2) koppies vir Sunlight™ skottelgoedopwasmiddel; en kontrole gereedskap wat bestaan het uit twee (2) bottels en twee (2) koppies. Die teikenbevolking vir die studie het bestaan uit voedingsbottels en -koppies vir suigelinge. Die analise vir die APC-kulture wat vergelyk is in die koppies vs. bottels om te bepaal of daar ’n beduidende verskil is tussen die gemiddelde bakterie-tellings, toon dat die gemiddelde bakterie-telling (op die In-skaal) is 6 cfu/ml en 9 cfu/ml vir die koppies en bottels respektiewelik. Die t-waarde is -1.17524. Aangesien die p-waarde 0.2595 is, is daar geen beduidende verskil gevind tussen die koppies en die bottels nie. Die E. coli-kulture is vergelyk in die koppies vs. bottels om te bepaal of daar ’n beduidende verskil tussen die gemiddelde bakterie-tellings is. Die uitslae wys dat die gemiddelde bakterie-telling (op die In-skaal) is 7cfu/ml en 7.6 cfu/ml vir koppies en bottels respektiewelik. Die t-waarde is -0.211902. Die p-waarde is 0.835237 en dus is daar geen beduidende verskil tussen koppies en bottels nie. Gevolgtrekking Die huidige studie toon dat daar geen beduidende verskil tussen die steriliseringsmetodes of tussen die gebruik van of bottels of koppies is nie. Dus, ’n studie met ’n groter steekproefgrootte word aanbeveel vir toekomstige navorsing. Aanbevelings Die navorser beveel aan dat toekomstige navorsers meer omvattende studies met ’n groter steekproefgrootte oor die onderwerp uitvoer. Studies met ’n groter steekproefgrootte sal veroorsaak dat die werklike verskille vanweë hul grootte genoegsaam sal wees, om beduidend te wees. Die gebruik van sonlig as ’n steriliseringsmetode vir die gereedskap van suigelinge word aanbeveel, aangesen dit beide tyd- en kostebesparend is. Sonlig is ’n goedkoop en maklik verkrygbare metode van sterilisasie; dus kan dit gebruik word deur gemeenskappe wat nie oor die nodige middele beskik nie, vanweë hul sosio-ekonomiese situasies.

published_or_final_version / Community Medicine / Master / Master of Public Health

O objetivo deste trabalho foi caracterizar astrovírus em amostras fecais coletadas de crianças com e sem diarréia, em São Paulo, Brasil, e divididas em dois grupos, EPM e HU, de acordo com a origem. A detecção foi realizada utilizando-se RT-PCR, com primers específicos. Os resultados para as amostras EPM mostram que 66/234 (28,2%) foram positivas para astrovírus. Para as amostras HU, 18/187 (9,6%) foram positivas. A genotipagem foi realizada com a técnica de nested/RT-PCR. De 66 amostras positivas (EPM), 19 (28,7%) foram caracterizadas como HAstV-1, 4 (6,0%) como HAstV-2, 2 (3,0%) como HAstV-3, 1 (1,5%) como HAstV-5 e 3 (4,5%) como HAstV-8. Das 18 positivas do HU, 1 (5,5%) amostra foi caracterizada como HAstV-1, 7 (38,8%) como HAstV-2 e 1 (5,5%) como HAstV-8. As amostras genotipadas em ambos os grupos foram submetidas ao seqüenciamento de nucleotídeos para confirmação dos resultados. Detecção e genotipagem de astrovírus em casos de diarréias pediátricas são técnicas são importantes e descrevem como esse vírus está circulando em São Paulo, Brasil. / The purpose of this study was to characterize astrovirus in faecal samples collected from children with and without diarrhea in São Paulo city, Brazil, and grouped into two distinct groups, EPM and HU. Detection was carried out using RT-PCR with specific primers. Results for EPM set showed that 66/234 (28,2%) were positive. In the HU set of samples, 18/187 (9,6%) were positive for astrovirus. Genotyping was carried out with nested/RT-PCR. Out of 66 astrovirus positive EPM samples, 19 (28,7%) were characterized as HAstV-1, 4 (6,0%) as HAstV-2, 2 (3,0%) as HAstV-3, 1 (1,5%) as HAstV-5 and 3 (4,5%) as HAstV-8. Among 18 astrovirus positive HU samples, 1 (5,5%) was characterized as HAstV-1, 7 (38,8%) as HAstV-2 and 1 (5,5%) as HAstV-8. Genotyped samples were confirmed by nucleotide sequencing. Detection and genotyping of astrovirus in pediatric diarrhea are important and describes how this virus is circulating in São Paulo, Brazil.

As gastroenterites ainda são um importante problema de saúde pública nos países em desenvolvimento e as Escherichia colí são as principais causas de diarréias. Adultos brasileiros apresentam anticorpos reativos com E. colí enteropatogênica (EPEC), que apresenta muitas semelhanças com E. colí enterohemorágica (EHEC) e pode ser responsável por complicações como a síndrome hemolítica urêmica e a colite hemorrágica. Ambas as bactérias apresentam um mecanismo patogênico comum: a formação da lesão \"attaching and effacing\" nos microvilus dos enterócitos, mediadas por fatores de virulência, como Intimina, Tir e as Esps. Os lipopolissacarídeos (LPS) são componentes da membrana externa das bactérias gram-negativas, incluindo E. colí. A infecção por EHEC 0157 resulta na produção de anticorpos séricos antiLPS 0157, que geralmente são indicativos de infecção recente. Neste trabalho, nós investigamos a presença de anticorpos séricos IgG e IgM anti-EHEC 0157:H7, EHEC 0111:H- e EPEC 0111:H- em adultos brasileiros, da Grande São Paulo. Amostras de soro foram coletadas de 200 adultos saudáveis, doadores de sangue, e um pool foi formado com 100 amostras. Anticorpos foram determinados em 100 amostras individuais por ELlSA com bactérias íntegras e anti-LPS para a determinação de anticorpos específicos anti-LPS 0111 e 0157 com o \"pool\" de soro como controle, em relação às concentrações de IgG e IgM totais determinadas simultaneamente. Os resultados foram submetidos a análise estatística. O repertório de anticorpos IgG e IgM para as três bactérias foi investigado por \"immunoblotting\" (18). A presença de anticorpos anti-bacterianos e anti-LPS foi confirmada. Existe uma correlação positiva entre os anticorpos reativos com as três bactérias, e entre os anticorpos anti-bactéria e anti-LPS. As concentrações de IgM anti-LPSs foi mais alta que IgG, o que está de acordo com o mecanismo imune esperado para um antígeno timo-independente. As concentrações de anti-LPS 0157 foram altas, apesar da baixa freqüência da bactéria 0157 em nosso meio. Os ensaios de 18 mostraram anticorpos reativos com bandas antigênicas sugestivas de fatores de virulência. A origem destes em nossa população poderia ser o contato com bactérias da microbiota ou do ambiente, de animais ou alimentos. Alternativamente, nossa população pode estar exposta à EHEC mais freqüentemente do que imaginado até agora. / Gastroenteritis is still an important public health problem in developing countries and Escherichia coli are frequent agents of diarrhea. Brazilian adults present antibodies reactive with the principal virulence factors of enteropathogenic E. coli (EPEC), which have many genetic and antigenic similarities with enterohemorrhagic E. coli (EHEC), that may be responsible for complications following diarrhea, as haemolytic uremic syndrome (HUS) and hemorrhagic colitis (HC). Both bacteria present a common pathogenic mechanism, with the formation of “attaching and effacing" lesion in microvilis’ enterocytes, mediated by virulence factors codified by the patogenicity island LEE, as Intimin, Tir e Esps. Lipopolysaccharides (LPS) are components of outer membrane and important virulence factors of Gram-negative bacteria including E. coli. The infection with EHEC O157 results in the production of serum antibodies to the O157 LPS antigens, and usually they are indicators of recent infection. In this work we investigated IgG and IgM serum antibodies reactive with EHEC O157:H7, EHEC O111:H- and EPEC O111:H- antigens in healthy Brazilian adults, living in São Paulo. Serum samples were collected from 200 healthy adults (blood donors) and a pool was formed with 100 samples. The antibody levels were determined by ELISA for 100 individual serum samples by means of a whole cell ELISA with the three bacteria and an anti-LPS ELISA for the determination of the concentrations of specific antibodies anti-LPS O111 and O157, using the serum pool as control, in relation to the total IgG and IgM concentrations determined simultaneously. The results were submitted to statistical analysis. The repertoire of IgG and IgM antibodies to the three bacteria was investigated by immunoblotting (IB). The presence of anti-bacterial and anti-LPS seric antibodies was confirmed. There is a positive correlation between the titers of antibodies reactive with the three bacteria and between anti-bacteria and anti-LPS antibodies. The concentrations of IgM anti-LPSs were significantly higher than IgG, which is in accordance with the immune mechanism expected to a thymus-independent antigen. Surprisingly the concentrations of anti-LPS O157 were high taking into account the low frequency of O157 bacteria isolation in our country. The IB assays showed the presence of antibodies, mainly IgG, reactive with many antigenic bands suggestive of virulence factors. The origin of anti-EHEC antibodies in our population could be the contact with microbiota or environment bacteria, and animal or human E. coli strains, pathogenic or not. Alternatively, our people may be exposed to EHEC more frequently than previously thought.

De um total de 187 amostras fecais coletadas no ambulatório do Hospital Universitário/USP, entre 1994 a 1996, 54 (28,9%) foram positivas para rotavírus. Entre as amostras caracterizadas por EGPA foram identificados nove perfis eletroforéticos longos, dois curtos e um tipo não usual. O subgrupo II e o sorotipo G2 foram os mais freqüentemente identificados. Foram caracterizadas três amostras com misturas de sorotipos. As amostras positivas e mais 163 amostras, coletadas em um laboratório particular, em 2000, foram genotipadas. Os genótipos G2P[4] e G1P[8] foram os mais freqüentes nos anos de 1994-1996 e G1P[8] e G9P[8], os mais freqüentes em 2000. Os genótipos G3 e G4 foram detectado em menor freqüência. No HU, 20 (38,5%) amostras foram caracterizadas como misturas de genótipos G e 16 (29,6%), como misturas de genótipos P; não foram identificadas misturas em 2000. Dezoito amostras foram caracterizadas como P[10] por RT-PCR mas a análise da seqüência de nucleotídeos mostrou uma homologia de 90,7 a 98,0% com a amostra padrão P[8]. / From 187 fecal samples collected from outpatients at Hospital Universitário (HU)/ USP, between 1994 to 1996, 54 (28,9%) were positive for rotavirus. Positive samples were submitted to electropherotyping, subgrouping, and G serotype. Electropherotypes were characterized as nine different long genome profiles, one short and one unusual profile. Subgroup II and G2 serotype were the most frequently found and three samples showed mixed serotypes. Rotavirus samples and an additional 163 positive fecal samples, collected in a private laboratory in 2000, were G and P genotyped. Genotypes G2P[4] and G1P[8] were the most frequently found in 1994-1996 and, in 2000, G1P[8] and G9P[8] were the most frequent. Genotype G3 and G4 were detected as minor strains in both years. For HU, G genotype mixtures were found in 20 (38.5%) samples and P mixtures were found in 16 (29.6%). No mixtures were identified in 2000. Nucleotide sequencing and phylogenetic analysis of 18 P[10] samples by RT-PCR showed 90.7 to 98.0% homology with the P[8] prototype.

Introdução: Norovírus é o principal agente viral de infecções gastrointestinais no mundo e sua alta infectividade gera aumento importante da demanda e dos custos para a saúde pública. Surtos ocorrem em ambientes fechados ou com aglomeração de pessoas. No município de São Paulo, o Norovírus tem se destacado como principal agente etiológico das gastroenterites. Objetivo: Descrever as noroviroses como causa de gastroenterite epidêmica no Município de São Paulo no período de 2010 a 2016. Métodos: Estudo transversal descritivo com dados dos sistemas de Vigilância Epidemiológica de surtos de gastroenterite e da Vigilância Sentinela Laboratorial do Rotavírus do Município de São Paulo entre os anos de 2010 a 2016. A definição de caso foi a identificação laboratorial do Norovírus como agente etiológico de surto de doença diarreica em todas as faixas etárias ou de casos esporádicos em menores de cinco anos internados em unidades sentinelas da Vigilância do Rotavírus. Os dados foram descritos segundo tempo, lugar e pessoa, por meio do Epi InfoTM versão 3.5.4 e ArcGIS versão 10.1. Resultados: O Norovírus foi associado a 20,4 por cento (68/334) dos surtos com pesquisa de agente realizada. Ocorrendo com maior frequência na região Norte, seguida das regiões Sul e Sudeste do Município de SP; principalmente em creches, domicílios e hospitais. Houve um predomínio de casos em crianças menores de 5 anos (47,2 por cento do sexo masculino; 28,6 por cento do sexo feminino) e em mulheres entre 20 a 49 anos (38,9 por cento). Na Vigilância Sentinela Laboratorial do Rotavírus do município de São Paulo, o Norovírus foi associado a 28,4 por cento (444/1565) dos casos menores de 5 anos. Os casos foram provenientes principalmente das regiões Norte e Sul, onde estão localizadas as duas unidades sentinelas. Verificou-se pico de ocorrência do Norovírus nos meses mais quentes. Destaca-se que o perfil das gastroenterites descrito foi fortemente influenciado pelas características da Vigilância Epidemiológica das Doenças de Transmissão Alimentar do Município de São Paulo. Conclusão: O Norovírus foi o principal agente etiológico de surtos de gastroenterite e de casos menores de 5 anos internados por diarreia aguda no Município de São Paulo. A vigilância das gastroenterites por Norovírus é importante para o estabelecimento de uma rede integrada entre diferentes estados e países que possibilitem o conhecimento da doença, planejamento de medidas de prevenção e controle e comunicação da informação. / Introduction: Norovirus is the major viral agent of gastroenteritis in the world and its high infectivity causes an important increase in demand and costs for public health. Outbreaks occur in closed environments or crowded settings. In the city of São Paulo, Norovirus has been highlighted as the main etiological agent of gastroenteritis. Objective: To describe the noroviruses as the cause of epidemic gastroenteritis in the city of São Paulo from 2010 to 2016. Methods: Descriptive cross-sectional study with data from the Epidemiological Surveillance systems of gastroenteritis outbreaks and Laboratory Sentinel Surveillance of Rotavirus of the city of São Paulo from 2010 and 2016. The case definition was the laboratory identification of Norovirus as the etiological agent of diarrheal disease outbreak in all age groups or sporadic cases in children under five years of age hospitalized in sentinel units of Rotavirus Surveillance. The data were described by time, place and person, using Epi InfoTM version 3.5.4 and ArcGIS version 10.1. Results: Norovirus was associated to 20.4 per cent (68/334) of outbreaks with agent identification performed. Occurred more frequently in the North region, followed by the South and Southeast regions of the city of São Paulo, especially in nurseries, residences and hospitals. A predominance of cases in children aged <5 years (47.2 per cent male, 28.6 per cent female) and in women aged 20 to 49 years (38.9 per cent) was observed. In the Laboratory Sentinel Surveillance of Rotavirus in the city of São Paulo, Norovirus was associated with 28.4 per cent (444/1565) of cases in children younger than 5 years old. The cases were mainly from the North and South regions, where the two sentinel units are placed. There was a peak of Norovirus occurrence in warmer months. It can be noticed that the profile of gastroenteritis described was strongly influenced by the characteristics of Foodborne Disease Surveillance of the city of São Paulo. Conclusion: Norovirus was the major etiological agent of gastroenteritis outbreaks and diarrhea cases under 5 years old hospitalized in the city of São Paulo. Surveillance of gastroenteritis caused by Norovirus is important for the establishment of an integrated network between different states and countries to contribute to disease knowledge, planning of prevention and control actions and distribution of information.

Introdução: Norovírus é o principal agente viral de infecções gastrointestinais no mundo e sua alta infectividade gera aumento importante da demanda e dos custos para a saúde pública. Surtos ocorrem em ambientes fechados ou com aglomeração de pessoas. No município de São Paulo, o Norovírus tem se destacado como principal agente etiológico das gastroenterites. Objetivo: Descrever as noroviroses como causa de gastroenterite epidêmica no Município de São Paulo no período de 2010 a 2016. Métodos: Estudo transversal descritivo com dados dos sistemas de Vigilância Epidemiológica de surtos de gastroenterite e da Vigilância Sentinela Laboratorial do Rotavírus do Município de São Paulo entre os anos de 2010 a 2016. A definição de caso foi a identificação laboratorial do Norovírus como agente etiológico de surto de doença diarreica em todas as faixas etárias ou de casos esporádicos em menores de cinco anos internados em unidades sentinelas da Vigilância do Rotavírus. Os dados foram descritos segundo tempo, lugar e pessoa, por meio do Epi InfoTM versão 3.5.4 e ArcGIS versão 10.1. Resultados: O Norovírus foi associado a 20,4 por cento (68/334) dos surtos com pesquisa de agente realizada. Ocorrendo com maior frequência na região Norte, seguida das regiões Sul e Sudeste do Município de SP; principalmente em creches, domicílios e hospitais. Houve um predomínio de casos em crianças menores de 5 anos (47,2 por cento do sexo masculino; 28,6 por cento do sexo feminino) e em mulheres entre 20 a 49 anos (38,9 por cento). Na Vigilância Sentinela Laboratorial do Rotavírus do município de São Paulo, o Norovírus foi associado a 28,4 por cento (444/1565) dos casos menores de 5 anos. Os casos foram provenientes principalmente das regiões Norte e Sul, onde estão localizadas as duas unidades sentinelas. Verificou-se pico de ocorrência do Norovírus nos meses mais quentes. Destaca-se que o perfil das gastroenterites descrito foi fortemente influenciado pelas características da Vigilância Epidemiológica das Doenças de Transmissão Alimentar do Município de São Paulo. Conclusão: O Norovírus foi o principal agente etiológico de surtos de gastroenterite e de casos menores de 5 anos internados por diarreia aguda no Município de São Paulo. A vigilância das gastroenterites por Norovírus é importante para o estabelecimento de uma rede integrada entre diferentes estados e países que possibilitem o conhecimento da doença, planejamento de medidas de prevenção e controle e comunicação da informação. / Introduction: Norovirus is the major viral agent of gastroenteritis in the world and its high infectivity causes an important increase in demand and costs for public health. Outbreaks occur in closed environments or crowded settings. In the city of São Paulo, Norovirus has been highlighted as the main etiological agent of gastroenteritis. Objective: To describe the noroviruses as the cause of epidemic gastroenteritis in the city of São Paulo from 2010 to 2016. Methods: Descriptive cross-sectional study with data from the Epidemiological Surveillance systems of gastroenteritis outbreaks and Laboratory Sentinel Surveillance of Rotavirus of the city of São Paulo from 2010 and 2016. The case definition was the laboratory identification of Norovirus as the etiological agent of diarrheal disease outbreak in all age groups or sporadic cases in children under five years of age hospitalized in sentinel units of Rotavirus Surveillance. The data were described by time, place and person, using Epi InfoTM version 3.5.4 and ArcGIS version 10.1. Results: Norovirus was associated to 20.4 per cent (68/334) of outbreaks with agent identification performed. Occurred more frequently in the North region, followed by the South and Southeast regions of the city of São Paulo, especially in nurseries, residences and hospitals. A predominance of cases in children aged <5 years (47.2 per cent male, 28.6 per cent female) and in women aged 20 to 49 years (38.9 per cent) was observed. In the Laboratory Sentinel Surveillance of Rotavirus in the city of São Paulo, Norovirus was associated with 28.4 per cent (444/1565) of cases in children younger than 5 years old. The cases were mainly from the North and South regions, where the two sentinel units are placed. There was a peak of Norovirus occurrence in warmer months. It can be noticed that the profile of gastroenteritis described was strongly influenced by the characteristics of Foodborne Disease Surveillance of the city of São Paulo. Conclusion: Norovirus was the major etiological agent of gastroenteritis outbreaks and diarrhea cases under 5 years old hospitalized in the city of São Paulo. Surveillance of gastroenteritis caused by Norovirus is important for the establishment of an integrated network between different states and countries to contribute to disease knowledge, planning of prevention and control actions and distribution of information.

Background & Objective: Acute gastroenteritis (AG) outbreak in elderly homes is common in Hong Kong, especially during the winter. Although mainly a self-limiting condition, the associated short-term as well as long-term medical and social costs can be extensive. This case-control study aims to investigate the hygienic risk factors related to infectious AG in elderly homes at both institutional and individual levels. Predictor variables under investigation include hand wash practice, infection control practice, routine institutional hygienic practice, food handling practice, and environmental factors such as the home setting, ventilation measures and isolation room setting. / Conclusions: This study found that 'sometimes or never wash hands after toilet' was a significant personal hygienic risk factor for AG transmission. This indicated that toilet may be the most susceptible place and hands are the most susceptible vehicle for AG transmission in Hong Kong elderly homes. A higher percentage of the NOHs had a more frequent routine cleaning practice than the OHs, demonstrating that routine cleaning practice may be an economical and an effective way to prevent AG infection. / Methods: All the elderly homes in the New Territories East were invited to take part in the study. A total of 34 homes and 2,995 residents were recruited in the study sample. The data collection period was from Dec 2007 to May 2009. Cases were notified within one week after a reported AG case, either by a report from the elderly home in question, the weekly check up with the New Territories East Community Geriatric Assessment Teams (NTE CGATs), regular contact with the elderly homes by the research assistant and case referrals from the Accident and Emergency Department from the Prince of Wales Hospital (PWH). One hundred and forty cases and 280 matched controls were recruited. For every AG case reported, two sex and age (within 5 years) and elderly home matched controls were selected. Structured questionnaires were conducted in face-to-face interviews in the elderly homes by trained interviewers. Information about the ventilation and the environmental hygiene of the elderly homes was collected by observation from the research team at the beginning of the study. Descriptive analysis was performed for the characteristics of cases and controls. Multivariate and multilevel logistic regression models were applied and odds ratios (ORs) were calculated for the potential hygienic risk factors. / Results: Multiple conditional logistic regression analysis revealed 'sometimes or never wash hands after toilet' OR:3.09 (95%CI: 1.28 -- 7.42) [ref gp: wash hands every time after toilet] was the major significant risk factor for AG in elderly homes, indicating the possible route of person-to-person transmission. Other significant risk factors included: Self-nutrition evaluation as 'not enough' (OR: 2.07; 95%CI: 1.05 -- 4.06), 'Being hospitalized in past month before the interview' (OR: 2.86; 95%CI: 1.16 -- 7.05), 'Simplified Barthel Index scored &lt;15" (OR: 2.63; 1.06 -- 6.53), and 'Alzheimer's' (OR: 2.75; 95% 1.18 -- 6.40). The institutional hygiene factors were investigated based on the descriptive analysis between the outbreak homes (OHs) and the non-outbreak homes (NOHs). The results indicated that the health worker (HW) to resident ratio was much lower in OHs than NOHs (50% OHs: 1:30-55 vs > 80% NOHs:1:10-29), and a higher percentage of the NOHs had a more frequent routine cleaning practice than the OHs. / Fung, Pui Kwan. / Adviser: Ho Suzanne Sutying. / Source: Dissertation Abstracts International, Volume: 73-02, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 182-206). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.