Introdução: A etiologia da otite média com efusão ainda não está completamente estabelecida, mas agentes infecciosos podem contribuir para sua patogênese. Demonstrou-se que a reação em cadeia da polimerase (PCR) é superior ao exame cultural para detectar espécies bacterianas. O conhecimento sobre a epidemiologia bacteriana da otite média com efusão em áreas geográficas distintas é essencial para a implementação de tratamentos racionais, quando necessários. Objetivos: Determinar a prevalência do Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis e Alloiococcus otitidis nas efusões de orelha média de crianças com otite média recorrente e otite média com efusão crônica que foram submetidas à miringotomia, comparar os resultados obtidos por cultura e PCR, comparar os achados bacteriológicos em crianças menores e maiores de dois anos e determinar o perfil de resistência à penicilina dos germes isolados. Métodos: Analisaram-se 128 amostras de efusões de orelha média de 75 crianças entre 11 meses e 9 anos e 4 meses de idade (média = 34,7 meses). Pacientes com otite média recorrente tinham efusão documentada por ≥ 6 semanas e aqueles com otite média com efusão crônica, por ≥3 meses. Os pacientes não tinham sinais de otite média aguda ou infecção do trato respiratório e não estavam sob antibioticoterapia no momento do procedimento. A aspiração do material foi realizada por timpanocentese, utilizando-se um coletor de Alden-Senturia. Os estudos bacteriológicos foram iniciados em menos de 15 minutos após a obtenção da efusão e uma parte da amostra foi armazenada a -20oC para análise posterior pela PCR. Utilizou-se um método de PCR simultânea para a detecção de quatro patógenos. A análise estatística foi efetivada com o teste χ2 de McNemar, teste χ2 com correção de Yates e teste exato de Fisher, quando apropriados. Resultados: Cultivaram-se bactérias em 32 (25,1%) das 128 amostras e os patógenos principais foram encontrados em 25 (19,6%). O A. otitidis não foi isolado em cultura. A PCR identificou bactérias em 110 (85,9%) das amostras, e os resultados positivos foram: 67 (52,3%) para A. otitidis, 50 (39,1%) para H. influenzae, 16 (12,5%) para S. pneumoniae e 13 (10,2%) para M. catarrhalis. Todas as amostras positivas por cultura foram positivas pela PCR, mas 85 (77,2%) das efusões com resultado positivo pela PCR foram negativas por cultura, para os germes estudados. A PCR foi significativamente mais sensível que a cultura (P<0,001). O S. pneumoniae foi encontrado mais freqüentemente em otite média recorrente do que em otite média com efusão crônica (P=0,038) e o H. influenzae foi encontrado mais vezes em crianças menores de dois anos (P=0,049). Quanto ao perfil de resistência, 100% das M. catarrhalis, 62,5% dos S. pneumoniae e 23% dos H. influenzae eram resistentes à penicilina. Conclusões: A prevalência das bactérias na otite média com efusão em um grupo de crianças brasileiras é semelhante àquelas relatadas em outros países, sendo o H. influenzae o mais encontrado dentre os patógenos principais da orelha média. Essa prevalência sugere que bactérias podem desempenhar um papel na patogênese da otite média com efusão. Os resultados mostram que a PCR é mais sensível na detecção de bactérias na efusão da orelha média, comparada com cultura, e é essencial para a identificação do A. otitidis. O elevado percentual de detecção do A. otitidis sugere mais investigações sobre sua atuação no início e no prolongamento de doenças da orelha média. O S. pneumoniae foi mais freqüente em otite média recorrente do que em otite média com efusão crônica e o H. influenzae foi mais encontrado em crianças menores de dois anos. A resistência à penicilina por parte do pneumococo e da moraxela é semelhante à relatada em outros países, ao passo que a produção de β-lactamase pelo hemófilo é mais baixa que aquela referida em bactérias isoladas em amostras de efusões de otite média com efusão. / Background: The etiology of otitis media with effusion is still unclear but infective agents may contribute to its pathogenesis. Polymerase chain reaction (PCR) has been shown to have a superior ability in detecting bacterial species, when compared to conventional culture methods. The knowledge of the bacteriological epidemiology of otitis media with effusion in different geographical areas is crucial for the implementation of rational treatment in selected cases. Objectives: To determine the prevalence of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Alloiococcus otitidis in the middle ear effusion of children with recurrent otitis media and chronic otitis media with effusion undergoing miryngotomy, to compare the results obtained by culture and PCR, to compare the bacteriological findings in children younger and older than two years of age, and to determine the susceptibility to penicillin of the bacterial isolates. Methods: A total of 128 middle ear effusion samples from 75 children aged 11 months to 9 years and 4 months (mean = 34.7 months) were analyzed. Patients with recurrent otitis media had documented middle ear effusion for ≥ 6 weeks, and chronic otitis media with effusion for ≥ 3 months. Patients had no signs of acute otitis media or respiratory tract infection and were not on antibiotics. Aspiration was done through tympanocentesis with an Alden-Senturia trap. Bacteriological studies were initiated in less than 15 minutes after acquisition of the effusion and a part of the sample was stored frozen at -20oC for latter PCR analysis. Multiplex PCR methods for the detection of four pathogens were used. Statistical analyses were done using McNemar´s χ2 test, χ2 test with Yates’ correction, and Fisher’s exact test, when appropriate. Results: Bacteria were cultured in 32 (25.1%) of the 128 samples and the major pathogens were found in 25 (19.6%). A. otitidis was not detected by culture. PCR yielded positive for bacteria in 110 (85.9%) of the samples and these positive PCR results were: 67 (52.3%) for A. otitidis, 50 (39.1%) for H. influenzae, 16 (12.5%) for S. pneumoniae, and 13 (10.2%) for M. catarrhalis. All the culture-positive samples were PCR-positive but 85 (77.2%) of the PCR-positive specimens were culture-negative. PCR was significantly more sensitive than culture (P<0.01). S. pneumoniae was more frequently found in samples from recurrent otitis media when compared to chronic otitis media with effusion (P=0.038) and H. influenzae was more prevalent in children younger than two years when compared to the older group (P=0.049). The resistance to penicillin was: M. catarrhalis = 100%; S. pneumoniae = 62.5% and H. influenzae = 23% of the isolates. Conclusions: The prevalence of bacteria in otitis media with effusion in a group of Brazilian children is similar to that reported from other countries, and H. influenzae is the most frequently found microorganism among the main middle ear pathogens. This prevalence suggests that bacteria may play a role in the pathogenesis of otitis media with effusion. Also PCR is more sensitive in detecting bacteria in the middle ear effusion, compared to conventional culture methods, and is essential for the detection of A. otitidis. The high recovery rate of A. otitidis warrants further investigation of its role in initiating or prolonging middle ear disease. S. pneumoniae was more frequently found in recurrent otitis media compared to chronic otitis media with effusion and H. influenzae was more prevalent in children younger than two years of age. Pneumococcal and moraxella´s resistance to penicillin is similar to but hemophillus’ β-lactamase production is lower than that reported from other countries when bacteria isolated from middle ear effusion samples of otitis media with effusion were analyzed.

Introdução: A etiologia da otite média com efusão ainda não está completamente estabelecida, mas agentes infecciosos podem contribuir para sua patogênese. Demonstrou-se que a reação em cadeia da polimerase (PCR) é superior ao exame cultural para detectar espécies bacterianas. O conhecimento sobre a epidemiologia bacteriana da otite média com efusão em áreas geográficas distintas é essencial para a implementação de tratamentos racionais, quando necessários. Objetivos: Determinar a prevalência do Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis e Alloiococcus otitidis nas efusões de orelha média de crianças com otite média recorrente e otite média com efusão crônica que foram submetidas à miringotomia, comparar os resultados obtidos por cultura e PCR, comparar os achados bacteriológicos em crianças menores e maiores de dois anos e determinar o perfil de resistência à penicilina dos germes isolados. Métodos: Analisaram-se 128 amostras de efusões de orelha média de 75 crianças entre 11 meses e 9 anos e 4 meses de idade (média = 34,7 meses). Pacientes com otite média recorrente tinham efusão documentada por ≥ 6 semanas e aqueles com otite média com efusão crônica, por ≥3 meses. Os pacientes não tinham sinais de otite média aguda ou infecção do trato respiratório e não estavam sob antibioticoterapia no momento do procedimento. A aspiração do material foi realizada por timpanocentese, utilizando-se um coletor de Alden-Senturia. Os estudos bacteriológicos foram iniciados em menos de 15 minutos após a obtenção da efusão e uma parte da amostra foi armazenada a -20oC para análise posterior pela PCR. Utilizou-se um método de PCR simultânea para a detecção de quatro patógenos. A análise estatística foi efetivada com o teste χ2 de McNemar, teste χ2 com correção de Yates e teste exato de Fisher, quando apropriados. Resultados: Cultivaram-se bactérias em 32 (25,1%) das 128 amostras e os patógenos principais foram encontrados em 25 (19,6%). O A. otitidis não foi isolado em cultura. A PCR identificou bactérias em 110 (85,9%) das amostras, e os resultados positivos foram: 67 (52,3%) para A. otitidis, 50 (39,1%) para H. influenzae, 16 (12,5%) para S. pneumoniae e 13 (10,2%) para M. catarrhalis. Todas as amostras positivas por cultura foram positivas pela PCR, mas 85 (77,2%) das efusões com resultado positivo pela PCR foram negativas por cultura, para os germes estudados. A PCR foi significativamente mais sensível que a cultura (P<0,001). O S. pneumoniae foi encontrado mais freqüentemente em otite média recorrente do que em otite média com efusão crônica (P=0,038) e o H. influenzae foi encontrado mais vezes em crianças menores de dois anos (P=0,049). Quanto ao perfil de resistência, 100% das M. catarrhalis, 62,5% dos S. pneumoniae e 23% dos H. influenzae eram resistentes à penicilina. Conclusões: A prevalência das bactérias na otite média com efusão em um grupo de crianças brasileiras é semelhante àquelas relatadas em outros países, sendo o H. influenzae o mais encontrado dentre os patógenos principais da orelha média. Essa prevalência sugere que bactérias podem desempenhar um papel na patogênese da otite média com efusão. Os resultados mostram que a PCR é mais sensível na detecção de bactérias na efusão da orelha média, comparada com cultura, e é essencial para a identificação do A. otitidis. O elevado percentual de detecção do A. otitidis sugere mais investigações sobre sua atuação no início e no prolongamento de doenças da orelha média. O S. pneumoniae foi mais freqüente em otite média recorrente do que em otite média com efusão crônica e o H. influenzae foi mais encontrado em crianças menores de dois anos. A resistência à penicilina por parte do pneumococo e da moraxela é semelhante à relatada em outros países, ao passo que a produção de β-lactamase pelo hemófilo é mais baixa que aquela referida em bactérias isoladas em amostras de efusões de otite média com efusão. / Background: The etiology of otitis media with effusion is still unclear but infective agents may contribute to its pathogenesis. Polymerase chain reaction (PCR) has been shown to have a superior ability in detecting bacterial species, when compared to conventional culture methods. The knowledge of the bacteriological epidemiology of otitis media with effusion in different geographical areas is crucial for the implementation of rational treatment in selected cases. Objectives: To determine the prevalence of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Alloiococcus otitidis in the middle ear effusion of children with recurrent otitis media and chronic otitis media with effusion undergoing miryngotomy, to compare the results obtained by culture and PCR, to compare the bacteriological findings in children younger and older than two years of age, and to determine the susceptibility to penicillin of the bacterial isolates. Methods: A total of 128 middle ear effusion samples from 75 children aged 11 months to 9 years and 4 months (mean = 34.7 months) were analyzed. Patients with recurrent otitis media had documented middle ear effusion for ≥ 6 weeks, and chronic otitis media with effusion for ≥ 3 months. Patients had no signs of acute otitis media or respiratory tract infection and were not on antibiotics. Aspiration was done through tympanocentesis with an Alden-Senturia trap. Bacteriological studies were initiated in less than 15 minutes after acquisition of the effusion and a part of the sample was stored frozen at -20oC for latter PCR analysis. Multiplex PCR methods for the detection of four pathogens were used. Statistical analyses were done using McNemar´s χ2 test, χ2 test with Yates’ correction, and Fisher’s exact test, when appropriate. Results: Bacteria were cultured in 32 (25.1%) of the 128 samples and the major pathogens were found in 25 (19.6%). A. otitidis was not detected by culture. PCR yielded positive for bacteria in 110 (85.9%) of the samples and these positive PCR results were: 67 (52.3%) for A. otitidis, 50 (39.1%) for H. influenzae, 16 (12.5%) for S. pneumoniae, and 13 (10.2%) for M. catarrhalis. All the culture-positive samples were PCR-positive but 85 (77.2%) of the PCR-positive specimens were culture-negative. PCR was significantly more sensitive than culture (P<0.01). S. pneumoniae was more frequently found in samples from recurrent otitis media when compared to chronic otitis media with effusion (P=0.038) and H. influenzae was more prevalent in children younger than two years when compared to the older group (P=0.049). The resistance to penicillin was: M. catarrhalis = 100%; S. pneumoniae = 62.5% and H. influenzae = 23% of the isolates. Conclusions: The prevalence of bacteria in otitis media with effusion in a group of Brazilian children is similar to that reported from other countries, and H. influenzae is the most frequently found microorganism among the main middle ear pathogens. This prevalence suggests that bacteria may play a role in the pathogenesis of otitis media with effusion. Also PCR is more sensitive in detecting bacteria in the middle ear effusion, compared to conventional culture methods, and is essential for the detection of A. otitidis. The high recovery rate of A. otitidis warrants further investigation of its role in initiating or prolonging middle ear disease. S. pneumoniae was more frequently found in recurrent otitis media compared to chronic otitis media with effusion and H. influenzae was more prevalent in children younger than two years of age. Pneumococcal and moraxella´s resistance to penicillin is similar to but hemophillus’ β-lactamase production is lower than that reported from other countries when bacteria isolated from middle ear effusion samples of otitis media with effusion were analyzed.

Durch die höhere Verfügbarkeit von Schnittbildverfahren in der Kleintiermedizin treten Flüssigkeitsansammlungen in der Bulla tympanica immer häufiger als Zufallsbefund bei Hunden, insbesondere bei brachyzephalen Rassen, auf. Aufgrund der fehlenden klinischen Symptome der Patienten ist eine akute Entzündung unwahrscheinlich. Daher bezeichnen neuere Studien diese Flüssigkeitsansammlung als primär sekretorische Otitis media (PSOM). In der vorliegenden Studie wurde untersucht, ob Hunde der Rassen Mops und Französische Bulldogge mit einer primär sekretorischen Otitis media einen konduktiven Hörverlust für das betroffene Ohr im Vergleich zu „Ohr gesunden“ Hunden ihrer Rasse zeigen. Des Weiteren wurde untersucht, ob Französische Bulldoggen signifikant häufiger eine primär sekretorische Otitis media als Hunde der Rasse Mops zeigen. Es wurden bei jeweils 41 Hunde der Rasse Mops und Französische Bulldogge, die zur chirurgischen Versorgung des Brachyzephalen Syndroms vorgestellt wurden, die frühen akustisch evozierten Potentiale (FAEP) abgeleitet. Des Weiteren erfolgte bei allen Patienten eine Computertomographie des Kopfes, eine Otoskopie und ggf. eine Punktion der Bulla tympanica inklusive einer Zytologie und bakteriologischen Untersuchung des Sekrets. In der vorliegenden Studie konnte kein konduktiver Hörverlust für die Hunde mit einer PSOM nachgewiesen werden. Es zeigte sich nur eine Verlängerung der Latenz der Welle I bei Vorliegen einer PSOM, jedoch ohne dass eine signifikant erhöhte Hörschwelle für diese Patienten festgestellt werden konnte. Als Ursache für die Verlängerung der Latenz der Welle I kommt sowohl die beschriebene Füllung der Bulla als auch ein stenotischer Gehörgang in Frage. Die Hörschwelle ist zwar bei den Hunden mit einer PSOM tendenziell höher als bei den Patienten ohne Füllung der Bullae, allerdings konnte hierfür kein signifikanter Unterschied nachgewiesen werden. Bei 40 % der untersuchten Patienten mit einseitiger PSOM konnte für beide Ohren dieselbe Hörschwelle bestimmt werden, so dass eine Füllung der Bulla tympanica nicht immer zu einer vorhersagbaren Veränderung der Hörschwelle führen muss. Eine mögliche Erklärung für diesen Umstand ist ein sensorineuraler Hörverlust, der durch eine chronische Entzündung des Mittelohres zustande kommt. Dies ist bisher nur in der Humanmedizin beschrieben und die Pathogenese ist noch unklar. Die Ergebnisse zeigen zusätzlich, dass Französische Bulldoggen signifikant häufiger von einer PSOM betroffen sind. Die Ursache für diese Häufung könnte ein dickerer weicher Gaumen der Französischen Bulldoggen im Vergleich zu Hunden der Rasse Mops sein, durch den es zu einer Funktionsstörung der Tuba auditiva kommen könnte. Beim Vergleich der Hunde ohne Vorliegen einer Füllung konnte festgestellt werden, dass Französische Bulldoggen eine höhere Hörschwelle und eine längere Latenz der Welle I als Hunde der Rasse Mops aufweisen. Bei beiden Rassen ist zusätzlich auffällig, dass die Hörschwelle um 30 dB höher liegt als bei Hunden anderer Rassen (SHIU et al. 1997). Alle brachyzephalen Hunde dieser Studie zeigen somit ein im Vergleich zu anderen Rassen vermindertes Hörvermögen, das aber durch eine PSOM nicht weiter verschlechtert wird. Der Symptomenkomplex Brachyzephalen Syndrom muss nach der vorliegenden Studie durch die Symptome vermindertes Hörvermögen und das Vorliegen einer PSOM ergänzt werden. Die klinischen Auswirkungen des Hörverlusts, wie etwa eine starke Anhänglichkeit, könnten bei Hunden dieser Rassen übersehen werden, da dies als gewünschter Charakterzug der Rassen interpretiert wird und nicht als mögliches Symptom einer Erkrankung wahrgenommen wird. / Because of the increased availability of cross-sectional imaging modalities in small animal medicine the incidental finding of material in the middle ear is more common, especially in brachycephalic dogs. Because the animals show no clinical signs, an acute inflammation is unlikely. Therefore recent studies term it as primary secretory otitis media (PSOM). The aim of the current study was to determine whether brachycephalic dogs with PSOM show a conductive hearing loss compared to brachycephalic dogs without changes in the middle ear. Additionally it was evaluated whether French bulldogs suffer from PSOM more frequent than pugs. BAER was recorded in 41 pugs and 41 French bulldogs, which were under general anesthesia because of the surgical correction of the brachycephalic syndrome. In all patients a computed tomography of the head, an otoscopy and if possible a myringotomy with aspiration of the fluid in the middle ear was performed. If fluid was available a cytological examination and a bacterial culture of the fluid was initiated. In the current study no conductive hearing loss was detected in brachycephalic dogs with PSOM. The latencies of wave I were increased in patients with PSOM, although the thresholds of hearing were not increased. The increased latencies of wave I can be explained by the fluid in the middle ear as well as the stenotic external ear canal. The thresholds of hearing in dogs with PSOM were tendentially higher than in dogs without fluid in the middle ear, but the correlation was not significant. In 40 % of the patients with PSOM in one ear the threshold of hearing in both ears is at the same level. Therefore there is no strict correlation between fluid in the middle ear and an increased threshold of hearing on the affected side. This could be explained by a sensorineural hearing loss caused by chronic inflammation of the middle ear. Until now a chronic inflammation as a cause of sensorineural hearing loss has only been described in human medicine, not in veterinary medicine. The pathogenesis is still unknown. The results show that French bulldogs suffer more frequently from the PSOM than pugs. French bulldogs usually show a bigger soft palate than pugs which could result in a dysfunction of the tuba auditiva. Hence the bigger soft palate could be causing the increased prevalence of PSOM in French bulldogs. Compared to pugs without PSOM, French bulldogs without PSOM show an increased latency of wave I as well as an increased threshold of hearing. Additionally for both breeds an increase in the threshold of hearing by 30 dB compared to normocephalic breeds could be detected (SHIU et al. 1997). It is remarkable that French bulldogs as well as pugs show a hearing loss without correlation to the PSOM. In conclusion hearing loss has to be added to the characteristic triad of symptoms of the brachycephalic syndrome. Clinical signs of hearing loss like loyalty to the owner could be misinterpreted especially in these breeds, since this is a favored behavior of these dogs.

Thesis (MSc)--University of Stellenbosch, 2006. / ENGLISH ABSTRACT: Strain ST4SA, isolated from soya beans, was identified as Enterococcus mundtii. BacST4SA, a bacteriocin produced by strain ST4SA inhibited the growth of Acinetobacter baumannii, Bacillus cereus, Clostridium tyrobutyricum, Enterococcus faecalis, Enterococcus faecium, Lactobacillus sakei, Propionibacterium spp., Streptococcus caprinus, Pediococcus sp., Listeria monocytogenes, Staphylococcus aureus, Streptococcus pneumoniae, and unidentified middle ear isolates A, BW, DW, F, G, and H. BacST4SA was active against Pseudomonas aeruginosa G, BG, I, J, B and E, although variable degrees of resistance were observed for some strains. BacST4SA is positively charged, hydrophobic, contains the YGNGV sequence in the N-terminal, a double-glycine processing site and a disulphide bridge, all of which is typical of a class IIa bacteriocin. The operon, which contains a structural-, ATP-dependent transporter- and immunity gene, is located on a 50-kb plasmid. The 58-amino acid prepeptide is homologous to mundticin KS, mundticin AT06 and bacteriocin QU 2, and differs from enterocin CRL35 by only two amino acids. The 674-amino acid ATP-dependent transporter, consisting of a peptidase C39B domain, an ABC-transporter and an ABC-DLP family domain, displayed 98.9% homology to mundticin KS and 99.25% to enterocin CRL35. The 98-amino acid immunity gene of bacST4SA is completely homologous to enterocin CRL35 and 96.9% to mundticin KS. BacST4SA is 3.950 kDa in size, based on electron spray mass spectrometry. The peptide was isolated from the cell-free supernatant, precipitated with 80% saturated ammonium sulphate, dialysed and freeze-dried to 1 638 400 AU (arbitrary units) per ml. No change in antimicrobial activity was recorded when bacST4SA was incubated in buffer ranging from pH 2 to 12, heated to 100 °C for 90 min and 121 °C for 20 min, and when incubated in the presence of Tween 20, Tween 80, Triton X-100, SDS, urea, EDTA, middle ear fluid and blood. Optimal levels of bacST4SA production (51 200 AU/ml) was recorded after 14 h of growth in MRS broth at 30°C. Maximum production (102 400 AU/ml) was recorded in modified MRS media supplemented with tryptone, yeast extract, a combination of tryptone and yeast extract, K2HPO4 (10.0 or 20.0 g/l), or with the addition of DL-6,8-thoictic acid, L-ascorbic acid, and thiamine, respectively. BacST4SA is bactericidal towards E. faecium HKLHS and bacteriostatic towards S. pneumoniae 40 and middle ear isolates F, BW and H. The peptide adsorbed maximal (94%) to S. pneumoniae 40, P. aeruginosa 25 and E. faecium HKLHS. BacST4SA forms pores in the cytoplasmic membrane of sensitive cells, leading to dissipation of the cell membrane and leakage of cytoplasmic material. BacST4SA was compared with various other antimicrobial treatment agents, and revealed similar to a higher activity towards a number of these agents. BacST4SA revealed a similar level of activity against E. faecium HKLHS and middle ear pathogens P. aeruginosa J and S. pneumoniae 27 when compared with tetracycline (30μg). However, bacST4SA revealed much higher activity when compared to nasal sprays, aminoglycosides, cephalosporins, fluoroquinolones, lincosamides, macrolides, nitroimidazole, penicillin, quinolones, sulfonamides, chloramphenicol, furanzolidone, fusidic acid, rifampicin, trimethoprim, trimethoprim-sulfamethoxazole and vancomycin when tested in vitro. / AFRIKAANSE OPSOMMING: Stam ST4SA, geïsoleer uit sojabone, is as Enterococcus mundtii geidentifiseer. BacST4SA, ‘n bakteriosien geproduseer deur stam ST4SA het die groei van Acinetobacter baumannii, Bacillus cereus, Clostridium tyrobutyricum, Enterococcus faecalis, Enterococcus faecium, Lactobacillus sakei, Propionibacterium spp., Streptococcus caprinus, Pediococcus sp., Listeria monocytogenes, Staphylococcus aureus, Streptococcus pneumoniae en ongeïdentifiseerde middeloor isolate A, BW, DW, F, G, en H geinhibeer. BacST4SA is aktief teen Pseudomonas aeruginosa stamme G, BG, I, J, B en E, alhoewel effense weerstand soms waargeneem is. BacST4SA het ‘n netto positiewe lading, is hidrofobies, bevat die YGNGV-volgorde in die N-terminaal, ‘n dubbel-glisien prosesserings setel en ‘n disulfied brug, kenmerkend van klas IIa bakteriosiene. Die operon, wat bestaan uit ‘n strukturele geen, ‘n ATP-afhanklike transport sisteem geen en ‘n immuniteits-geen, is op ‘n 50 kb plasmied gelokaliseer. Die voorloper peptied (58 aminosure lank), is homoloog aan mundticin KS, mundticin AT06 en bakteriosien QU 2 en verskil van enterocin CRL35 met slegs twee aminosure. Die ATP-afhanklike transporter (674 aminosure lank) bestaan uit ‘n peptidase C39B domein, ‘n ABC-transporter en ‘n ABC-DLP tipe domein en is 98.9% homoloog aan mundticin KS and 99.25% aan enterocin CRL35. Die immuniteits-geen (98 aminosure lank) van bacST4SA is ten volle homoloog aan enterocin CRL35 en 96.9% homoloog aan mundticin KS. BacST4SA is 3.950 kDa groot, gebaseer op elektrosproei-massa spektrometrie. Die peptied is uit selvrye supernatant geïsoleer, met 80% versadigde ammonium sulfaat gepresipiteer, gedialiseer en gevriesdroog tot ’n finale konsentrasie van 1 638 400 AE (arbitrêre eenhede) per ml. Geen verandering in antimikrobiese aktiwiteit is waargeneem tydens inkubasie van bacST4SA in buffer van pH 2 tot 12, tydens verhitting (100 °C vir 90 min en 121 °C vir 20 min) en tydens inkubasie in die teenwoordigheid van Tween 20, Tween 80, Triton X-100, SDS, ureum, EDTA, middeloor vloeistof en bloed. Optimale vlakke van bacST4SA produksie (51 200 AE/ml) is na 14 h groei in MRS media by 30°C waargeneem. Maksimale vlakke van die peptied (102 400 AE/ml) is geproduseer in gemodifiseerde MRS medium, aangevul met triptoon, gisekstrak, ‘n kombinasie van triptoon en gisekstrak, K2HPO4 (10.0 of 20.0 g/l), of met byvoeging van DL-6,8-thioktiensuur, L-askorbiensuur, en tiamien onderskeidelik. BacST4SA is bakteriosidies teenoor E. faecium HKLHS en bakteristaties teenoor S. pneumoniae 40 en middeloor isolate F, BW en H. Die peptied adsorbeer optimaal (94%) aan S. pneumoniae 40, P. aeruginosa 25 en E. faecium HKLHS. BacST4SA vorm porieë in die selmembraan van sensitiewe selle en lei tot vernietiging van die selmembraan en lekkasie van die sitoplasma inhoud. In vergelykende studies het bacST4SA ‘n soortgelyke en selfs hoër antimikrobiese aktiwiteit teenoor ‘n aantal bekende antimikrobiese middels getoon. Die aktiwiteit van bacST4SA is soortgelyk aan dié van tetrasiklien (30μg) in toetse teen E. faecium HKLHS en middeloor patogene P. aeruginosa J en S. pneumoniae 27. BacST4SA het egter in ’n in vitro vergelyking met neussproeie, aminoglisiedes, cephalosporiene, fluoroquinolone, lincosamides, makroliede, nitroimidazole, penisilien, quinolone, sulfonamide, chloramphenicol, furanzolidone, fusiensuur, rifampisien, trimethoprim, trimethoprim-sulfamethoxazool en vankomisien ‘n baie hoër aktiwiteit teen patogene getoon.

Chronic otitis media (OM) is an archetypal complex disease, which is particularly prevalent in childhood. Epidemiological data suggest high heritability for disease susceptibility, but previous genetic association studies have had methodological flaws, and none have specifically focused on chronic OM phenotypes. Mouse models represent one way to ascertain candidate loci for human association testing. A number of mouse models of middle ear inflammation have been reported, but many susceptibility loci remain undiscovered. I demonstrate that oto-endoscopy is a robust and scalable phenotyping platform for OM in the mouse, and discuss its value in new model discovery. Chronic OM is also a feature of trisomy HSA21 (Down Syndrome). Through an interrogation of the mouse library of segmental trisomy models of Down Syndrome, I identify a critical trisomic region for chronic otitis media. This region may underlie OM susceptibility in Down Syndrome, but could also contribute to disease susceptibility in non-syndromic disease. Mouse models can also be used to interrogate disease mechanisms. Our previous work has shown that the chronically inflamed middle ear is hypoxic, and that hypoxia signalling is a potential therapeutic target. Exploiting the Junbo mouse model, I demonstrate that surgical ventilation of the Junbo ear improves inflammation, and that this is associated with loss of hypoxia signalling. I present preliminary results from transcript analyses of human middle ear effusions showing marked upregulation of hypoxia signalling. A systematic review of existing mouse models suggests that the loci FBXO11, EVI1, SMAD2, and TGIF1 are good candidates genes for human association testing. I detail recruitment and collection of DNA from families in the UK where a child is undergoing grommet insertion. Association testing using a variant of the transmission disequilibrium test shows susceptibility associated with polymorphisms at FBXO11, and possibly also SMAD2 and TGIF1.

This qualitative research enabled the participants (three young people, aged nine to fourteen years, and seven mothers) to explain their experience of living with long-term Glue Ear. As far as I could ascertain this is the first research in which young people and their mothers have led the researcher in exploring their experience. This innovative research illustrates the effects of the condition at home and school. It provides insight into participants’ views of interactions with healthcare and educational professionals. Photovoice is usually used with groups, but in this research it was successfully adapted to enable individual participants to reflect on their experience. The data was analysed using Interpretative Phenomenological Analysis (IPA) to keep the focus on each participant’s words. Innovatively combining these two methodologies provided depth to both the data gathering and the data analysis. Five healthcare and educational professionals were interviewed to understand their perspective. The contribution of this research is that the methodology enabled young people and their mothers to reflect on their experience and lead the researcher in understanding the impact Glue Ear had on their day-to-day life. It highlighted the need for educational professionals to receive more formal training about Glue Ear’s potential educational and social impacts and to potentially adapt their teaching and learning practices to better support affected students. Young people and their parents demonstrated that they have valuable information to contribute to decisions about their healthcare and education requirements and would welcome the opportunity to work collaboratively with such professionals.

Devido a intensa relação entre a fissura labiopalatina e a deficiência auditiva, este trabalho teve como objetivo desenvolver e analisar a eficácia de um ambiente virtual de transmissão de informações sobre fissura labiopalatina e deficiência auditiva para profissionais, com a utilização da Teleducação Interativa. Como metodologia, foi desenvolvido um ambiente de aprendizagem no formato de um blog contendo informações sobre fissura labiopalatina e deficiência auditiva, englobando conteúdo específico direcionado a incidência, etiologia e tipos de fissura; deficiência auditiva; adaptação e seleção de AASI, funcionamento da tuba auditiva, otite média, etiologia da alteração de orelha média nos indivíduos com fissura labiopalatina, influência da otite no desenvolvimento da linguagem; tratamento para otite média: intervenção cirúrgica e acompanhamento sistemático e adaptação de AASI. Este blog foi disponibilizado na internet, sendo realizada a avaliação, por meio de dois questionários, o primeiro é o Emory (Emory University Rollins School of Public Health 1998), traduzido para o português (Bastos 2011), e o segundo questionário corresponde à avaliação específica do conteúdo do blog. A avaliação foi realizada por 75 profissionais do HRAC-USP, Bauru e outros, por demanda espontânea. O blog foi elaborado e disponibilizado na web pelo endereço: www.fissuraeaudicao.wordpress.com. No período analisado, de agosto de 2010 a 25 de junho de 2012, foram obtidas 13.746 visitas, sendo a página mais visitada sobre Fissura labiopalatina tipos de fissura, com 4.064 acessos. Além do Brasil diversos países acessaram o conteúdo do blog. O termo de busca mais utilizado foi tuba auditiva. A maioria dos avaliadores do blog (70,7%) era formada por profissionais Fonoaudiólogos. Destes, 89% consideraram o blog como excelente e 11% como adequado. Quanto ao conteúdo, dos 70 profissionais que responderam a este questionário como um todo, 69% consideraram excelente, 27% adequado e 4% consideraram o conteúdo pobre, sendo estes últimos fonoaudiólogos. Assim, um ambiente virtual, em formato de blog, contendo informações sobre fissura labiopalatina e deficiência auditiva para profissionais da área da saúde, foi desenvolvido e avaliado pela maioria dos juízes como excelente. / Due to the intense relationship between cleft palate and hearing loss, this work had as objective to develop and to analyze the effectiveness of a virtual environment of transmission of information on cleft palate and hearing loss for professionals, with the use of Interactive Teleducation. As methodology, an environment of learning in blog format was developed, containing information about cleft lip and palate and hearing loss, encompassing specific content directed to the incidence, etiology and types of cleft lip and palate; hearing loss; selection and adaptation of hearing aids, functioning of the auditory tube, otitis media, etiology of the alteration of the middle ear in individuals with cleft palate, influence of the otitis in the language development; treatment for otitis media: surgical intervention and systematic follow up and adaptation of hearing aids. This blog was made available on the Internet, and the evaluation was made through two questionnaires, being the first one Emory (Emory University Rollins School of Public Health 1998), translated into Portuguese (Bastos 2011), and the second questionnaire corresponds to the specific evaluation of the blog content. The evaluation was carried through by 75 professionals of the HRAC-USP-Bauru and others, through spontaneous demand. The blog was elaborated and made available on the web on the address: www.fissuraeaudicao.wordpress.com. In the analyzed period from August 2010 to June 25, 2012, there were 13.746 visits, being the most visited page about cleft lip and palate - types of cleft lip and palate, with 4.064 accesses. In addition to Brazil, several countries accessed the contents of the blog. The most used search term was auditory tube. The majority of the blog evaluators (70.7%) consisted of professional Speech and Language Pathologists. Of these, 89% considered the blog excellent and 11% adequate. In regard to the content, out of the 70 professionals who had answered this questionnaire as a whole, 69% had considered it excellent, 27% adequate and 4% considered the content poor, being these last ones Speech and Language Pathologists. Thus, a virtual environment, in blog format, containing information on cleft palate and hearing loss for health professionals was developed and evaluated by the majority of the judges as excellent.

A fissura labiopalatina é um indicador de risco para alterações de orelha média e estas podem prejudicar o desenvolvimento de habilidades auditivas como, por exemplo, a atenção, que é essencial para o aprendizado de novas habilidades, inclusive da comunicação oral e escrita. O estudo do processo atencional na população com fissura labiopalatina é algo recente e pouco explorado na literatura específica consultada, assim, este trabalho poderá contribuir com novos subsídios na área, uma vez que teve como objetivos: a) verificar o desempenho de crianças com essa anomalia craniofacial em dois testes, o THAAS e o teste dicótico de dígitos (etapa de escuta direcionada, que avaliaram processos de atenção auditiva); b) comparar o resultado com um grupo sem fissura labiopalatina e; c) verificar a associação entre os dois testes aplicados. Fizeram parte do estudo 55 crianças, de ambos os gêneros, na faixa etária de 7 anos a 7 anos e 11 meses, que foram distribuídas em dois grupos: a) grupo controle, formado por crianças sem fissura labiopalatina; b) grupo experimental, formado por crianças com fissura labiopalatina. Para ambos os grupos, o processo de avaliação constituiu-se em: aplicação de um questionário; bateria de testes auditivos convencionais; aplicação do teste da habilidade de atenção auditiva (THAAS) (FENIMAN, 2004) e do teste dicótico de dígitos etapa de escuta direcionada (SANTOS; PEREIRA, 1997). Foi possível observar que o desempenho do grupo com fissura labiopalatina foi inferior ao do grupo controle em todos os tipos de resposta do THAAS e diferença estatisticamente significativa ocorreu para o decréscimo da vigilâ (p=0,014). No teste dicótico de dígitos - etapa de escuta direcionada, o grupo com fissura labiopalatina apresentou porcentagens de acerto inferiores ao grupo controle, tanto para a orelha direita quanto para a orelha esquerda. A análise estatística mostrou interação estatisticamente significante entre grupo e gênero (p=0,026). Ao comparar o THAAS com o teste dicótico de dígitos, foi possível observar que existe associação entre os testes, mas, essa associação mostrou-se muito baixa (R²=0,27). As crianças com fissura labiopalatina apresentaram desempenho no THAAS inferior àquelas sem esta anomalia craniofacial, apenas para o decréscimo da vigilância. No teste dicótico de dígitos - etapa de escuta direcionada, somente as crianças do gênero feminino com fissura labiopalatina obtiveram índices de acerto inferiores às do grupo controle. Uma baixa associação foi verificada entre o THAAS e o teste dicótico de dígitos - etapa de escuta direcionada, permitindo supor que habilidades diferentes são responsáveis pelo desempenho nos dois testes. / Cleft lip and palate indicates risk to alterations in the middle ear. These risks may impair the development of some hearing abilities, such as attention, which is essential to learn new abilities, including oral and written communication. Studies on attention process with the population with cleft lip and palate are recent and not widely found in literature. Therefore, this study can contribute to the area. The aims of this study were to examine children with this craniofacial anomaly through two tests: The SAAAT and the Dichotic Digit test - directed hearing stage which evaluated the hearing attention processes; to compare the results with a group without cleft lip and palate; to verify the association between the two tests. 55 children, both genders, aged 7 to 7 years and 11 months old were divided in two groups to be submitted to the study. Experimental group consisted of children with cleft lip and palate and Control group consisted of children without it. Both groups were assessed through a questionnaire, conventional hearing tests battery, the Sustained Auditory Attention Ability test (SAAAT) (Feniman, 2004), and the Dichotic Digit test - directed hearing stage (Santos; Pereira, 1997). Experimental group showed lower performance than the control group in all kinds of answers of the SAAAT and, significant difference regarding decrease in vigilance (p=0,014). In the Dichotic Digit test the experimental group showed lower percentages of right answers than the control group, not only for the right ear but also for the left ear. Statistic analysis showed significant interaction between group and gender (p=0,026). When compared, the SAAAT and the Dichotic digit test had low association (R²=0,27). Experimental Group presented lower performance in the SAAAT only at the vigilance decrease. At the Dichotic Digit test female children with cleft lip and palate presented lower scores of right answers than the Control group. The SAAAT and The Dichotic Digit test - directed hearing stage - were not closely associated. Thus, it is possible to assume that different abilities are responsible for the performance in both tests.

Introdução: A otite média com efusão recorrente nos primeiros cinco anos de vida é considerada um indicador de risco para alterações no processamento auditivo central (PAC). No entanto, não foram encontrados estudos que avaliassem o PAC em sujeitos com otite média crônica. Em virtude da gravidade e cronicidade da doença, destaca-se a relevância deste estudo. Objetivo: Investigar e analisar o impacto da otite média crônica não colesteatomatosa (OMCNC) no processamento auditivo central e em alguns aspectos neuropsicológicos de adolescentes. Métodos: Trata-se de um estudo observacional, transversal controlado, no qual foram recrutados 34 adolescentes de 12 a 18 anos com diagnóstico de otite média crônica não colesteatomatosa oriundos de um ambulatório de atendimento a indivíduos com otite média crônica de um hospital do sul do país. Um grupo controle de 34 adolescentes pareados por idade, sexo, renda familiar e escolaridade materna foi estabelecido para o estudo. A avaliação dos indivíduos da amostra foi realizada por meio dos seguintes procedimentos: anamnese, audiometria tonal liminar, logoaudiometria, bateria de avaliação comportamental do processamento auditivo central (Masking Level Difference- MLD, Synthetic Sentence Identification com mensagem competitiva ipsilateral- SSI/MCI, Random Gap Detection Test- RGDT, Duration Pattern Sequence- DPS e Teste Dicótico de Dígitos- DD) e alguns subtestes do Instrumento de Avaliação Neuropsicológica Breve Neupsilin (tarefas de atenção, memória de trabalho e função executiva). Resultados: Em relação ao PAC, verificou-se diferença estatisticamente significativa entre as médias observadas entre os grupos estudo e controle em todos os testes realizados: MLD (p= 0,001), SSI em relação sinal/ruído de 0 dB (p< 0,001) em ambas as orelhas, SSI em relação sinal-ruído de -15dB (p< 0,001) em ambas as orelhas, RGDT (p< 0,001), DPS (p=0,002), DD na orelha direita (p= 0,001) e DD na orelha esquerda (p=0,003). Nos sub testes do Neupsilin, observou-se diferença significativa entre os grupos nas seguintes tarefas: repetição de sequência de dígitos (p= 0,011), ordem ascedente de dígitos (p= 0,025), span auditivo de sentenças (p= 0,002) e fluência verbal fonêmica (p= 0,001). Conclusões: Foi constatado impacto no PAC e nos subtestes de atenção, memória e função executiva em adolescentes com otite média crônica não colesteatomatosa. Desta forma, ressalta-se a necessidade de encaminhamento de sujeitos com a doença para avaliação e intervenção terapêutica. / Introduction: Otitis media with recurrent effusion in the first five years of life is considered an indicator of risk for alterations in central auditory processing (CAP). However, no studies were found evaluating CAP in subjects with chronic otitis media. Due to the severity and chronicity of the disease, the relevance of this study stands out. Objective: To investigate and analyze the impact of chronic non-cholesteatomatous otitis media on central auditory processing and on neuropsychological aspects in adolescents. Methods: This was a cross-sectional, observational study in which 34 adolescents aged 12 to 18 years with a diagnosis of chronic non-cholesteatomatous otitis media from an outpatient clinic for individuals with chronic otitis media were recruited from a hospital in southern Brazil. A control group of 34 adolescents matched by age, sex, family income and maternal schooling was established for study. The evaluation of the subjects was performed through the following procedures: anamnesis, tonal threshold audiometry, logoaudiometry, behavioral evaluation of central auditory processing battery (Masking Level Difference- MLD, Synthetic Sentence Identification with ipsilateral competitive message- SSI / MCI, Random Gap Detection Test- RGDT, Duration Pattern Sequence- DPS and Digit Dichotic Test- DD) and some subtests of the Neupsilin Brief Neuropsychological Assessment Instrument (attention, working memory and executive function). Results: In relation to CAP, a statistically significant difference was observed between the means observed between the study and control groups in all the tests performed: MLD (p = 0.001), SSI in 0 dB signal-to-noise ratio in both ears (p <0.001), SSI in -15dB signal-to-noise ratio in both ears (p = 0.001), RGDT (p < 0,001), DPS (p=0,002), DD in the right ear (p = 0.001) and DD in the left ear (p= 0.003). In the Neupsilin sub-tests, a significant difference was observed between groups in the following sub-tests: digit sequence repetition (p = 0.011), digit ascending order (p = 0.025), auditory sentence span (p = 0.002), and Phonemic verbal fluency (p = 0.001). Conclusions: Impact on PAC and on subtests of attention, memory and executive function were observed in adolescents with chronic non-cholesteatomatous otitis media. Thus, the need to refer subjects with the disease for evaluation and therapeutic intervention is emphasized.

Objetivos: Verificar a ocorrência de problemas auditivos em crianças com seqüência de Robin, entre 1 à 3 anos de idade, com fissura palatina, não submetidas a palatoplastia. Comparar a freqüência de problemas auditivos em crianças com seqüência de Robin isolada (grupo I), com crianças com outras anomalias associadas a seqüência de Robin (grupo II), na mesma faixa etária, não submetidas a palatoplastia. Resultados: Os achados demonstram que crianças do grupo II são submetidas à internação em 100%, para 56% do grupo I. Apenas 26% dos pais ou cuidadores do grupo I e 48% do grupo II, apresentaram algum tipo de queixa auditiva. A queixa mais relatada foi à otalgia (70% no grupo I e 86% no grupo II), precedida da otorréia em 20% no grupo I e 14% no grupo II e apenas 10% relatou duvidas quanto à perda auditiva observadas apenas no grupo I. Predominou a curva tipo B em ambos os grupos (76% no grupo I e 83% no grupo II) com ausência de reflexos acústicos (93% no grupo I e 96% no grupo II) considerando as medidas de imitância acústica. A perda auditiva foi encontrada em 33% no grupo I, sendo 78% de perda auditiva de grau leve, no grupo II perda auditiva ocorreu em 81%, 47% de grau leve, 41% de grau moderado e 12% de grau severo. Conclusões: Lactentes com SR apresentam riscos para problemas audiológicos devido a possibilidade de hopsitalizações nos primeiros meses de vida, alterações de orelha média devido a fissura palatina e a presença de obstrução respiratória característica da desta anomalia. Os cuidadores apresentam dificuldades em observar sintomas ou características que evidenciem possibilidade de perda auditiva. Lactentes com SR isolada apresentam perda auditiva de grau leve, o aumento da gravidade da perda auditiva pode estar relacionado a fatores como síndromes genéticas e outras anomalias congênitas. A audiometria de reforço visual e as medidas de imitância acústica são instrumentos eficazes na detecção de possíveis problemas audiológicos em lactentes com SR. A avaliação audiológica de ser acrescentada no protocolo de tratamento de lactentes com SR, complementando a reabilitação, com atenção especial em casos do síndrome genética, principalmente a síndrome de Stickler. O atraso na cirurgia de palato não está proporcionando prejuízos à audição em crianças com SRI isolada. / Objectives: To investigate the occurency of hearin problems in children with Robin Sequence (RS) aged 1 to 3 years old with cleft palate not submitted to palatoplasty. To compare the frequency of hearing problems in children with RS only (group 1), children with other anomalies associated to RS (group 2), at the some age, not submitted to palatoplasty. Results: The findings demonstrate that 100% of children in group 2 are submitted to hospital admission compared to 56% in group 1. Only 26% of parents or caretakers of group 1 and 48% in group 2 were able to describe some kind of hearing complaints. The most frequent complaints were otalgia (70% in group 1 and 14% in group 2) and only 10% demonstrated doubts related to hearing loss observed only in group 1. In both groups the type B curve prevailed (76% in group 1 and 83% in group 2) considering timpanometry measurements. The hearing loss was detected in 33% of children in group 1, where 78% of the hearing loss were mild. In group 2 the hearing loss occurred in 81% of children, where 47% were mild,41% moderate and 12% were severe. Conclusions: Children with RS showed risks to hearing problems due to the possibility of hospitalization in the first months of life, alterations in middle ear due to cleft palate and the presence of respiratory obstruction characterized by this anomaly. Parents and caretakers show difficulties to observe symptoms or characteristics that prove the possibility of hearing loss. Children with RS only showed mild hearing loss, the increasing gravity of hearing loss could be related to factors such as genetic syndromes and othe congenit anomalies. The visual refoircement audiometry and the timpanometry are efficient instruments to detect possible hearing problems in children with RS. The audiologic findings must be added to the treatment protocol in children with, supplementing the rehabilitation, with special attention in genetic syndromes, specially the Stickler syndrome. The delay on palate surgery is not cousing damages on hearing in children with RS only.