Enfermedad invasiva por Haemophilus influenzae: reporte de 14 casos luego de un año de iniciada la pandemia por COVID-19 / Invasive Haemophilus influenzae disease: A report of 14 cases one year after the COVID-19 pandemic outbreak

El Haemophilus influenzae (Hi) causa enfermedad invasiva (EI). Se distinguen cepas capsuladas, como el serotipo b (Hib), y cepas no tipificables (HNT). Al año de declarada la pandemia por COVID-19, observamos un aumento de casos. Se describen las características clínico-epidemiológicas de niños con EI por Hi internados en el hospital (julio 2021-julio 2022). Hubo 14 casos; 12 previamente sanos. Aislamientos: Hib (n = 6), Hi serotipo a (n = 2), HNT (n = 5), 1 no se tipificó. Mediana de edad: 8,5 meses (RIC 4-21). Manifestaciones: meningitis (n = 5), neumonía (n = 6), celulitis (n = 2), artritis (n = 1). Nueve presentaron vacunación incompleta para Hib. Observamos un incremento de EI por Hi de 2,5 veces respecto a años previos. Estos datos sugieren el resurgimiento de Hib por la caída de las coberturas de vacunación y porque otras cepas de Hi no b están en aumento.

Haemophilus influenzae (Hi) causes invasive disease. There are encapsulated strains, such as serotype b (Hib), and non-typeable strains (NTHi). One year after the outbreak of the COVID-19 pandemic, the number of cases increased. In this report we describe the clinical and epidemiological characteristics of children hospitalized with invasive Hi disease (July 2021-July 2022). There were 14 cases; 12 were previously healthy children. Isolations: Hib (n = 6), Hi serotype a (n = 2), NTHi (n = 5); 1 case was not typified. Median age: 8.5 months (IQR: 4­21). Manifestations: meningitis (n = 5), pneumonia (n = 6), cellulitis (n = 2), arthritis (n = 1). Incomplete Hib immunization was observed in 9 children. Invasive Hi disease increased 2.5 times from previous years. These data suggest the reemergence of Hib due to a decline in vaccination coverage and an increase in other non-b-type Hi serotypes.

Estado de vacunación en pacientes con riesgo de enfermedades invasoras por bacterias capsuladas en un hospital pediátrico de Buenos Aires / Vaccination status in patients at risk for invasive disease with encapsulated bacteria at a children's hospital in the City of Buenos Aires

Introducción. El Ministerio de Salud contempla vacunas específicas para personas con riesgo elevado de infecciones invasoras por bacterias capsuladas (BC). En la actualidad se desconoce el cumplimiento del programa. El objetivo fue evaluar el estado de vacunación para BC en ≤ 18 años con factores de riesgo. Población y métodos. Estudio observacional, analítico, mediante encuesta a padres de ≤ 18 años con VIH, asplenia y/o déficit de complemento que concurrieron al vacunatorio de un hospital pediátrico de octubre de 2020 a septiembre de 2021. Se recabaron datos sociodemográficos y clínicos. Se evaluó el estado de vacunación para BC: neumococo, meningococo y Haemophilus influenzae b (Hib), calendario regular y antigripal. Se administró la escala de reticencia a la vacunación (ERV): rango 10-50. Se analizó la asociación entre las variables estudiadas y la vacunación para BC mediante regresión logística (OR, IC95%). Se utilizó la base datos REDCap® y STATA vs14®. Resultados. Participaron 104 sujetos, media 9,9 años (DE 4,4). Asplenia: 91,3 %, VIH: 7,6 % y déficit de complemento: 0,9 %. Nivel socioeconómico: pobreza relativa: 38,4 %, seguido por clase media: 37,5 %. Estado de vacunación completa para meningococo: 45 %, neumococo: 42 %, Hib: 97 %. El 77,9 % tenía al día el calendario regular y el 61,5 %, el antigripal. Media ERV: 41,9 (DE 3,2). No se encontraron asociaciones significativas entre las variables y el estado de vacunación para BC. Conclusiones. Un elevado porcentaje no tenía vacunación completa para BC, tampoco el calendario regular y antigripal. La confianza en la vacunación de los cuidadores fue elevada.

Introduction. The Ministry of Health has established specific vaccines for people at high risk for invasive infections with encapsulated bacteria (EB). There is currently no information about compliance with the vaccination schedule. Our objective was to assess EB vaccination status in subjects ≤ 18 years with risk factors. Population and methods. Observational, analytical study with a survey to parents of subjects aged ≤ 18 years with HIV, asplenia and/or complement deficiency attending a vaccination center at a children's hospital between October 2020 and September 2021. Sociodemographic and clinical data were collected. Their vaccination status for the EB pneumococcus, meningococcus, and Haemophilus influenzae type b (Hib), their regular vaccination and flu vaccination schedules were assessed. The vaccine hesitancy scale (VHS) was administered: range 10­50. The association between the study variables and EB vaccination was analyzed using logistic regression (OR, 95% CI). The REDCap® database and the STATA® v.14 software were used. Results. A total of 104 subjects participated; mean age: 9.9 years (SD: 4.4). Asplenia: 91.3%, HIV: 7.6%, and complement deficiency: 0.9%. Socioeconomic level: relative poverty: 38.4%, followed by middle class: 37.5%. Complete vaccination status: meningococcal vaccine 45%, pneumococcal vaccine: 42%, Hib: 97%. The regular vaccination and flu vaccination schedules were up-to-date in 77.9% and 61.5% of cases, respectively. Mean VHS score: 41.9 (SD: 3.2). No significant associations were observed between variables and EB vaccination status. Conclusions. A high percentage of subjects had not completed neither their EB vaccination nor their regular or their flu vaccination schedules. Caregivers' confidence in vaccines was high.

Espondilodiscitis por haemophilus parainfluenzae: un reporte de caso / Spondylodiscitis due to haemophilus parainfluenzae: a case report

INTRODUCCIÓN Haemophilus parainfluenzae (HP) es un cocobacilo gram negativo y un patógeno oportunista. Rara vez se asocia a infecciones vertebrales o musculoesqueléticas, y está muy poco reportado en la literatura. PRESENTACIÓN DELO CASO Una mujer de 45 años, sana, que presentaba un historial de dos semanas de lumbalgia progresiva, fiebre, coriza y congestión nasal, y que tenía discitis intervertebral causada por HP, confirmada por dos hemocultivos positivos y hallazgos progresivos de resonancia magnética (RM) de columna lumbar. Los hallazgos de la RM fueron atípicos, y consistían en un absceso del psoas y pequeñas colecciones de líquido epidural e intraespinal anterior asociadas con espondilodiscitis. El diagnóstico inicial se retrasó debido a que la RM inicial no reveló hallazgos que sugirieran un proceso infeccioso. El tratamiento consistió en un ciclo prolongado de administración intravenosa seguida de antibióticos orales, lo que finalmente produjo una buena respuesta clínica. DISCUSIÓN Y CONCLUSIÓN El HP es un patógeno muy raro en la espondilodiscitis. No obstante, debe tenerse en cuenta, especialmente en pacientes que presentan lumbalgia y fiebre y/o bacteriemia por microorganismos gram negativos. El estudio inicial debe incluir una RM de la columna con contraste. Aunque es poco común, la espondilodiscitis y un absceso del psoas pueden presentarse concomitantemente. Los antibióticos prolongados son el pilar del tratamiento.

INTRODUCTION Haemophilus parainfluenzae (HP) is a gram-negative coccobacillus and an opportunistic pathogen. It is rarely associated with spinal- and musculoskeletal-site infections, and very little reported in the literature. CASE PRESENTATION An otherwise healthy, 45-year-old woman who presented with a two-week history of progressive low back pain, fever, coryza and nasal congestion, was found to have intervertebral discitis caused by HP, confirmed by two positive blood cultures and progressive lumbar spine magnetic resonance imaging (MRI) findings. The MRI findings were atypical, consisting of a psoas abscess and small anterior epidural and intraspinal fluid collections associated with spondylodiscitis. The initial diagnosis was delayed because the initial MRI failed to reveal findings suggestive of an infectious process. The treatment consisted of a long course of intravenous followed by oral antibiotics, ultimately yielding a good clinical response. DISCUSSION AND CONCLUSION Haemophilus parainfluenzae is a very rare pathogen in spondylodiscitis. Nonetheless, it should be considered, especially in patients presenting with low back pain and fever and/or gram negative bacteremia. The initial work-up should include contrast-enhanced MRI of the spine. Although rare, spondylodiscitis and a psoas abscess can present concomitantly. Prolonged antibiotics are the mainstay of treatment.

Caracterización del polirribosilribitol fosfato por cromatografía de exclusión molecular de alta eficacia con detección ultravioleta / Characterization of polyribosylribitol phosphate by high efficiency size exclusion chromatography with ultraviolet detection

Haemophilus influenzae tipo b es un importante patógeno del hombre causante de varias de las enfermedades invasivas en niños menores de cinco años, contra el cual fueron autorizadas las vacunas glicoconjugadas a partir del polirribosilribitol fosfato. Quimi-Hib® es la primera y única vacuna contra este patógeno que utiliza el polisacárido obtenido por síntesis química. El Ingrediente Farmacéutico Activo es producido por el Centro de Ingeniería Genética y Biotecnología y se obtiene a partir de su conjugación al toxoide tetánico. En el presente reporte se hizo una caracterización del polirribosilribitol fosfato mediante la técnica de cromatografía de exclusión molecular de alta eficacia con detección ultravioleta a 215 nm. En el estudio se evaluaron tres lotes y se determinó el perfil de elución en una columna SuperdexTM 75 10/300 GL Increase con un porciento de pureza de 77,42 ± 8,97 y una masa molar promedio de 7.381 Da ± 210,93. La principal impureza presente en el polirribosilribitol fosfato es el dimetilsulfóxido, disolvente utilizado en la reacción de activación con el éster N-hidroxisuccinimidilo del ácido β-maleimidopropiónico. El polirribosilribitol fosfato se purificó por filtración con un Amicon Ultra-15 de 2.000 Da hasta una pureza de 99,1 por ciento y se conjugó al toxoide tetánico. El rendimiento de la reacción de conjugación con el polisacárido purificado fue de 30,0 por ciento 1,77 el cual no muestra diferencias significativas con el control que fue 33,7 por ciento ± 3,57 demostrándose que el dimetilsulfóxido no afecta el desempeño de la reacción de conjugación(AU)

Haemophilus influenzae type b is an important human pathogen causing some invasive diseases in children less than five years of age. Glycoconjugate vaccines based on polyribosylribitol phosphate have been licensed against this bacterium. Quimi-Hib® is the first and only vaccine against this pathogen using the chemically synthesized polysaccharide. The Active Pharmaceutical Ingredient is produced by the Center for Genetic Engineering and Biotechnology and is obtained from its conjugation to tetanus toxoid. In the present report a characterization of polyribosylribitol phosphate was performed by high performance molecular exclusion chromatography with ultraviolet detection at 215 nm. Three batches were evaluated in the study and the elution profile was determined on a SuperdexTM 75 10/300 GL Increase column with a purity percentage of 77.42 ± 8.97 and an average molecular weight of 7,381 Da ± 210.93. The main impurity present in polyribosylribitol phosphate was dimethylsulfoxide, the solvent used in the activation reaction with N-hydroxysuccinimidyl ester of β-maleimidopropionic acid. Polyribosylribitol phosphate was purified by filtration using a 2,000 Da cut-off Amicon Ultra-15 to a purity of 99.1 percent and conjugated to tetanus toxoid. The yield of the conjugation reaction with the purified polysaccharide was 30.0 percent ± 1.77 which shows no significant difference with the control which was 33.7 percent ± 3.57 demonstrating that dimethylsulfoxide does not affect the performance of the conjugation reaction(AU)

Glässer's disease in swine from Northeastern Brazil / Doença de Glässer em suínos no Nordeste do Brasil

Glässer's disease is an important infectious disorder of swine caused by Haemophilus parasuis. Although well recognized in most regions of Brazil, outbreaks of Glässer's disease have not been described in Northeastern region. For this reason, three municipalities of the Pernambuco State were visited in order to identify histories of high mortality in growing and finishing pigs. The main clinical signs consisted of dry cough, apathy, fever, anorexia, paresis, muscle tremors, motor incoordination, seizures leading to high mortality rates. Nine pigs were necropsied, and fragments of the nervous system, organs of the abdominal and thoracic cavities were collected for histological analysis. In addition, lung and brain fragments were used for DNA extraction and molecular testing by real-time Polymerase Chain Reaction (PCR). Grossly, the main lesions consisted of petechial hemorrhages or ecchymosis on the skin of the face, abdomen, forelimbs, and hind limbs. The main severe lesions consisted of hydropericardium, hemopericardium, fibrinous pericarditis and pleuropneumonia. Microscopically, pericarditis, epicarditis and subepicardial myocarditis, followed by a moderate to severe multifocal pleuropneumonia, fibrinosuppurative and necrotizing were the most frequent lesions observed. Real-time PCR amplified H. parasuis infB gene in all samples analyzed, confirming the presence of this etiologic agent.(AU)

A doença de Glässer é uma importante enfermidade infecciosa de suínos causada pela bactéria Haemophilus parasuis. Embora bem reconhecida na maioria das regiões do Brasil, surtos de doença de Glässer não têm sido descritos na região Nordeste. Por este motivo, três regiões do Estado de Pernambuco foram visitadas com o objetivo de se identificar históricos de alta mortalidade em leitões e suínos em fase de terminação. Nove suínos foram necropsiados e fragmentos do sistema nervoso, órgãos das cavidades abdominal e torácica foram coletados para análise histopatológica. Além disso, fragmentos de pulmão e cérebro foram utilizados para extração de DNA e realização de teste molecular por meio da Reação em Cadeia da Polimerase (PCR) em tempo real. Os principais sinais clínicos consistiram em tosse seca, apatia, febre, anorexia, paresia, tremores musculares, incoordenação motora e convulsões levando a altas taxas de mortalidade. As lesões macroscópicas mais severas consistiam em petéquias e equimoses na pele da face, abdome, membros anteriores e posteriores, além de hidropericárdio, hemopericárdio, pericardite fibrinosa e pleuropneumonia. Microscopicamente, pericardite, epicardite e miocardite subepicárdica, seguidas de pleuropneumonia multifocal moderada a grave, fibrino-supurativa e necrosante foram as lesões mais frequentes observadas. A PCR em tempo real amplificou o gene infB de H. parasuis em todas as amostras analisadas, confirmando a presença deste agente etiológico.(AU)

Effect of coinfection with influenza virus and bacteria on host damage / Efecto de la coinfección por virus de la influenza y bacterias en el daño al hospedero

Abstract Background: Influenza virus infection is often complicated by a bacterial infection, with this coinfection causing severe pneumonia. If not timely treated, the disease can cause death. Objective: To demonstrate, in animal models, that coinfection with influenza virus and bacteria that affect the respiratory tract causes multisystemic damage. Method: Six groups of mice were formed: a control group, one infected with the influenza virus, two infected with bacteria: Haemophilus influenzae and Streptococcus pneumoniae, respectively; and two co-infected with influenza virus and Haemophilus influenzae or Streptococcus pneumoniae, respectively. Results: Of the six groups of mice, only the group co-infected with influenza virus and Streptococcus pneumoniae showed damage to thoracic and abdominal organs. A decrease in serum cytokine levels was found in all study groups, which was more pronounced in the co-infected mice. Conclusions: The groups of mice infected with Streptococcus pneumoniae or influenza virus alone showed no damage, which indicates that coexistence of these infections caused the damage in the group of co-infected mice.

Resumen Antecedentes: La infección por el virus de la influenza con frecuencia se complica con una infección bacteriana, coinfección que provoca cuadros graves de neumonía, la cual puede ocasionar la muerte si no es tratada en forma oportuna. Objetivo: Demostrar en modelos animales que la coinfección por el virus de la influenza y bacterias que afectan el tracto respiratorio ocasiona daño multisistémico. Método: Se formaron seis grupos de ratones: un grupo control, uno infectado de virus de la influenza, dos infectados de bacterias: Haemophilus influenzae y Streptococcus pneumoniae, respectivamente; y dos coinfectados de virus de la influenza y Haemophilus influenzae y Streptococcus pneumoniae, respectivamente. Resultados: De los seis grupos de ratones, solo en el grupo coinfectado de virus de la influenza y Streptococcus pneumoniae se observó daño en órganos torácicos y abdominales. En todos los grupos se encontró disminución de los niveles séricos de las citocinas, mayor en los ratones coinfectados. Conclusiones: Los grupos de ratones infectados solo de Streptococcus pneumoniae o el virus de la influenza no presentaron daños, lo cual indica que la coexistencia de estas infecciones fue la que ocasionó el daño en el grupo de ratones coinfectados.

Invasive non-type b haemophilus influenzae disease: report of eight cases / Doença invasiva causada por haemophilus influenzae não tipo b: relato de oito casos

ASTRACT Objective: To describe eight cases of invasive non-type b Haemophilus influenzae disease in children admitted to Hospital de Clínicas of Universidade Estadual de Campinas. Cases description: In 2015, there were eight cases of invasive non-type b H. influenzae disease. We tested the ampicillin sensitivity and beta-lactamase production of the strains identified and performed the genotyping. Molecular typing was determined by Pulsed-Field Gel Electrophoresis. Four patients were diagnosed with bacteremia; in two cases, H. influenzae was detected in the pleural fluid, and two patients had meningitis. Patients with comorbidities represented 37.5% of cases. Except for the strain of one patient - not sent to the reference laboratory -, all were ampicillin-sensitive and non-beta-lactamase-producing. Genotyping identified four non-capsular, one type c, and two type a strains. Molecular typing ruled out nosocomial transmission since all serotypes were distinct regarding genotype. Comments: The rise in cases of invasive non-type b H. influenzae infection was real. There was no nosocomial transmission, and we found no justification for the increase. These data indicate the need for surveillance to correctly diagnose, monitor, and understand the spectrum of non-type b H. influenzae disease.

ABSTRACT Objetivo: Descrever oito casos de doença invasiva por Haemophilus influenzae não tipo b em crianças internadas no Hospital de Clínicas da Universidade Estadual de Campinas. Descrição dos casos: Em 2015, ocorreram oito casos de doença invasiva por H. influenzae não tipo b. Nas cepas identificadas, testou-se a sensibilidade à ampicilina e a produção de betalactamase, e realizou-se a genotipagem. A tipagem molecular foi feita por Pulsed Field Gel Electrophoresis. Em quatro pacientes, o diagnóstico foi de bacteremia; em dois casos, H. influenzae foi identificado em líquido pleural, e dois pacientes tiveram meningite. Comorbidades foram encontradas em 37,5% dos pacientes. Com exceção da cepa de um dos pacientes (que não foi enviada ao laboratório de referência), todas eram sensíveis à ampicilina e não produtoras de betalactamase. A genotipagem identificou quatro cepas não capsulares, uma cepa tipo c e duas cepas tipo a. A tipagem molecular descartou a transmissão intra-hospitalar, já que todos os sorotipos eram distintos quanto ao genótipo. Comentários: O aumento dos casos de infecção invasiva por H. influenzae não tipo b foi real. Não houve transmissão intra-hospitalar e não foi encontrada justificativa para o aumento. Esses dados indicam a necessidade de vigilância para diagnosticar corretamente, monitorar e entender o espectro da doença causada por H. influenzae não tipo b.

Colloidal gold immunochromatographic strip for rapid detection of Haemophilus influenzae / 生物工程学报

To establish a novel colloidal gold immunochromatography assay (GICA) for rapid, sensitive and accurate detection of Haemophilus influenzae infection by using the outer membrane protein P6 as detection target. First, the linear antigen epitope located in the extracellular domain of the P6 protein (GenBank accession number: AGH02799) was predicted by bioinformatics analysis. The region (62-75 aa of the protein) with strong antigen specificity was chosen and synthesized. Two rabbits were then immunized by the polypeptides (14 aa) for production of polyclonal antibodies. Then, the recombinant P6 proteins were also obtained to produce polyclonal antibodies. Finally, based on the two antibodies, a novel colloidal GICA for detection of Haemophilus influenzae infection was established and the specificity, sensitivity, repeatability and stability of this method were evaluated. At the same time, the method was tested in clinical simulation, and the plate culture method was used to verify its accuracy. The test strip for Haemophilus influenzae infection was successfully prepared. The detection limit of the test strip was as low as 1×105 CFU/mL and the whole process can be completed within 15 minutes. The strip specifically recognized Haemophilus influenzae and did not react with nine of other common respiratory pathogens such as Streptococcus pneumoniae, Moraxella catarrhalis, Mycoplasma pneumonia, and Legionella pneumophila. And the strips could be stored at 25 °C for at least 6 months without losing sensitivity or specificity. The coincidence rate between the results of 200 clinical samples and the plate culture method was 90.5%. Haemophilus influenzae protein P6, which possessed a high degree of surface antigen accessibility and antigencity, could be used as a marker for Haemophilus influenzae detection. The immunochromatographic colloidal gold test strip which bears the features of rapidity, convenience and sensitivity provides a unique tool for the on-site surveillance and diagnosis of Haemophilus influenzae infection in clinical test.

A Review of Traditional Vaccine-Preventable Diseases and the Potential Impact on the Otolaryngologist

Abstract Introduction A majority of otolaryngologists have not had direct experience with many vaccine-preventable diseases since the creation of national vaccination programs. Despite the elimination of endemic transmission of some of these diseases in the United States, outbreaks can occur anywhere and still pose a threat to public health around the world. Recent outbreaks and changing trends in exemption rates indicate that it is important for physicians to maintain a working knowledge of how these diseases present and of the recommended treatment guidelines. Objectives This review will evaluate the current state of vaccination rates, vaccine exemption rates and disease incidence in the United States and in the world. It will also examine the clinical presentation and treatment recommendations of these diseases. Data Synthesis United States estimated vaccination rates, vaccine exemption rates and vaccine-preventable disease incidences were obtained from data compiled by the Centers for Disease Control and Prevention. World vaccination rates and disease incidences were obtained from the World Health Organization databases, which compile official figures reported by member states. A PubMed literature review provided information on the current state of vaccination exemptions and outbreaks in the United States. Conclusion Vaccination and vaccine exemption rates continue to put the United States and many areas of the world at risk for outbreaks of vaccine-preventable diseases. Clinical guidelines should be reviewed in the event of a local outbreak.

Haemophilus influenzae no tipificable: mecanismo de destrucción del biofilm / Non-typeable Haemophilus influenzae: mechanism of biofilm destruction

INTRODUCCIÓN: Haemophilus influenzae tiene como hábitat el aparato respiratorio humano. No existe vacuna contra la variante no tipificable (NTHi) de este, por lo que se ha mostrado una tendencia al incremento de casos en los últimos años. La patogenicidad de NTHi se debe a su habilidad para formar biofilms, por lo que se pretende analizar mecanismos de destrucción del biofilm formado por NTHi. METODOLOGÍA: Se realizó una revisión bibliográfica mediante consultas en las bases de datos, Hinari, MEDLINE y el motor de búsqueda PubMed. Se tomaron en consideración artículos originales, ensayos clínicos y artículos de revisión bibliográfica dando prioridad a aquellos publicados en los últimos 5 años. DESARROLLO: NTHi tiene la capacidad de formar biofilms por medio de proteínas del pili y elementos estabilizadores del ADN extracelular, entre los más importantes están: DNA BII, pili tipo IV, Nucleasa extracelular 2019 y adhesinas de alto peso molecular. El uso de chalconas, EDTA y ADN como quelante de cationes, antisueros contra DNBII y maquinaria luxS más anticuerpos contra Pili IV han demostrado que pueden ser útiles para la erradicación del biofilm. CONCLUSIÓN: Se proponen dos tratamientos novedosos que podrían contribuir en la eliminación del biofilm formado por NTHi, uno es el uso de EDTA y otro los anticuerpos dirigidos a proteínas esenciales en la formación y adhesión del biofilm. Es necesario profundizar en otros estudios con estas propuestas terapéuticas para determinar su uso en el área clínica en un futuro.

INTRODUCTION: Haemophilus influenzae has the human respiratory system as habitat. There's no vaccine against the non-typeable group (NTHi), as a consequence there has been an increase in the number of cases in the past few years. The pathogenesis of NTHi is caused by its ability to form biofilms, for this reason we pretend to analyze the destruction mechanisms of biofilms formed by NTHi. METHODOLOGY: A review was made using the databases Hinari, MEDLINE and PubMed. Original articles, clinical trials and reviews that had been published in the last five years were taken in count. RESULTS: NTHi has the capacity to form biofilms through pilus proteins and extracellular DNA stabilizers; among the most important we have: DNA BII, type IV pilus, 2019 extracellular nuclease and adhesins of high molecular weight. The use of chalcone, EDTA, DNA as cation chelant, antiserum against DNA BII and luxS plus antibodies against Pili IV has shown potential to eradicate biofilms. DISCUSSION: Two new treatments, that could contribute to the removal of biofilms formed by NTHi, are proposed: one of them is the use of EDTA and the other one is the use of antibodies against essential proteins that the bacteria uses in the formation and adhesion of biofilms. Never the less, it is necessary to deepen more in studies about these therapeutic alternatives to determine their use in the clinical area in the future.

Infection by Streptococcus pneumoniae in children with or without radiologically confirmed pneumonia / Infecção por Streptococcus pneumoniae em crianças com ou sem pneumonia radiologicamente confirmada

Abstract Objective: Community-acquired pneumonia is an important cause of morbidity in childhood, but the detection of its causative agent remains a diagnostic challenge. The authors aimed to evaluate the role of the chest radiograph to identify cases of community-aquired pneumonia caused by typical bacteria. Methods: The frequency of infection by Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis was compared in non-hospitalized children with clinical diagnosis of community acquired pneumonia aged 2-59 months with or without radiological confirmation (n = 249 and 366, respectively). Infection by S. pneumoniae was diagnosed by the detection of a serological response against at least one of eight pneumococcal proteins (defined as an increase ≥2-fold in the IgG levels against Ply, CbpA, PspA1 and PspA2, PhtD, StkP-C, and PcsB-N, or an increase ≥1.5-fold against PcpA). Infection by H. influenzae and M. catarrhalis was defined as an increase ≥2-fold on the levels of microbe-specific IgG. Results: Children with radiologically confirmed pneumonia had higher rates of infection by S. pneumoniae. The presence of pneumococcal infection increased the odds of having radiologically confirmed pneumonia by 2.8 times (95% CI: 1.8-4.3). The negative predictive value of the normal chest radiograph for infection by S. pneumoniae was 86.3% (95% CI: 82.4-89.7%). There was no difference on the rates of infection by H. influenzae and M. catarrhalis between children with community-acquired pneumonia with and without radiological confirmation. Conclusions: Among children with clinical diagnosis of community-acquired pneumonia submitted to chest radiograph, those with radiologically confirmed pneumonia present a higher rate of infection by S. pneumoniae when compared with those with a normal chest radiograph.

Resumo Objetivo: Avaliar o papel do raios X de tórax na identificação de casos de pneumonia adquirida na comunidade (PAC) causada por agentes bacterianos. Métodos: A frequência de infecção por Streptococcus pneumoniae, Haemophilus influenzae e Moraxella catarrhalis em crianças com PAC não hospitalizadas foi comparada com a presença de confirmação radiológica da pneumonia (n = 249 crianças com pneumonia radiologicamente confirmada e 366 crianças com raios X de tórax normal). Infecção por S. pneumoniae foi diagnosticada com base na resposta sorológica a pelo menos uma dentre oito proteínas pneumocócicas investigadas (aumento ≥ 2 vezes nos níveis de IgG em relação a Ply, CbpA, PspA1 e 2, PhtD, StkP-C e PcsB-N ou aumento≥ 1,5 vez em relação aPcpA). Infecção por H. influenzae e M. catarrhalis foi definida por aumento ≥ 2 vezes nos níveis de IgG específica a antígenos de cada agente. Resultados: Crianças com pneumonia radiologicamente confirmada apresentaram maior taxa de infecção pelo pneumococo. Além disso, a presença de infecção pneumocócica foi um fator preditor de pneumonia radiologicamente confirmada, o que aumenta sua chance de detecção em 2,8 vezes (IC 95%: 1,8-4,3). O valor preditivo negativo do raios X normal para a infecção por S. pneumoniae foi 86,3% (IC95%: 82,4%-89,7%). Não houve diferença nas frequências de infecção por H. influenzae e M. catarrhalis entre crianças com PAC com ou sem confirmação radiológica. Conclusão: Crianças com diagnóstico clínico de PAC submetidas a um raios X de tórax que apresentam confirmação radiológica têm maior taxa de infecção por S. pneumoniae comparadas com as crianças com raios X normal.

Antimicrobial susceptibility patterns of Brazilian Haemophilus parasuis field isolates / Perfil de susceptibilidade antimicrobiana de isolados clínicos brasileiros de Haemophilus parasuis

Haemophilus parasuis is the etiological agent of Glässer's disease (GD), an ubiquitous infection of swine characterized by systemic fibrinous polyserositis, polyarthritis and meningitis. Intensive use of antimicrobial agents in swine husbandries during the last years triggered the development of antibiotic resistances in bacterial pathogens. Thus, regular susceptibility testing is crucial to ensure efficacy of different antimicrobial agents to this porcine pathogen. In this study, 50 clinical isolates from South Brazilian pig herds were characterized and analyzed for their susceptibility to commonly used antibiotic. The identification and typing of clinical isolates was carried out by a modified indirect hemagglutination assay combined with a multiplex PCR. The susceptibility of each isolate was analyzed by broth microdilution method against a panel of 21 antimicrobial compounds. We found that field isolates are highly resistance to gentamycin, bacitracin, lincomycin and tiamulin, but sensitive to ampicillin, clindamycin, neomycin, penicillin, danofloxacin and enrofloxacin. Furthermore, an individual susceptibility analysis indicated that enrofloxacin is effective to treat clinical isolates with the exception of those classified as serovar 1. The results presented here firstly demonstrate the susceptibility of Brazilian clinical isolates of H. parasuis to antimicrobials widely used by swine veterinary practitioners and strengthen the need to perform susceptibility test prior to antibiotic therapy during GD outbreaks. In addition, because only six antimicrobial drugs (28.6%) were found effective against field isolates, a continuous surveillance of the susceptibility profile should be of major concern to the swine industry.(AU)

Haemophilus parasuis é o agente etiológico da doença de Glässer (GD), um processo infeccioso que acomete suínos e que se caracteriza por poliserosites fibrinosas sistêmicas, poliartrites e meningites. O uso intensivo de agentes antimicrobianos na produção de suínos, durante os últimos anos, tem disparado a seleção de cepas bacterianas resistentes a antibióticos. Desta maneira, a avaliação rotineira de susceptibilidade torna-se crucial para assegurar a correta seleção de um antimicrobiano eficaz contra este patógeno. Neste estudo, analisou-se a susceptibilidade antimicrobiana de 50 isolados clínicos de H. parasuis procedentes de granjas localizadas na região sul do Brasil. A identificação e tipificação dos isolados clínicos foi realizada através de uma PCR multiplex combinada com o teste de hemaglutinação indireta modificada. A susceptibilidade de cada isolado foi analisada pelo método de microdiluição em caldo utilizando-se um painel composto por 21 agentes antimicrobianos. Os resultados deste estudo indicam que as cepas clínicas de H. parasuis apresentam alta resistência à gentamicina, bacitracina, lincomicina e tiamulina, no entanto, são susceptíveis a ampicilina, clindamicina, neomicina, penicilina, enrofloxacina e danofloxacina. A análise de susceptibilidade realizada dentro de cada grupo de cepas de um mesmo sorovar indicou que a enrofloxacina é o antibiótico mais efetivo para tratar todos isolados clínicos com exceção daqueles pertencentes ao sorovar 1. Em termos gerais, neste trabalho, demonstra-se o perfil de susceptibilidade de isolados clínicos de H. parasuis aos antimicrobianos comumente utilizados pelos médicos veterinários especialistas em suínos, e reforça-se a necessidade da realização de testes de susceptibilidade antes do início da terapia com antibióticos durante surtos de DG. Além disso, como somente seis antimicrobianos (28.6%) foram efetivos contra os isolados clínicos, uma vigilância contínua do perfil de susceptibilidade aos antimicrobianos deve ser de grande preocupação para a indústria de suínos.(AU)

Caracterización clínica de la meningitis aguda de la comunidad causada por Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae y Listeria monocytogenes en adultos en el Hospital Carlos van Buren entre los años 2013 y 2016 / Clinical characteristics of community-acquired acute meningitis caused by Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae and Listeria monocytogenes in adults in Carlos van Buren Hospital between 2013-2016

INTRODUCCIÓN: la meningitis bacteriana es una enfermedad infecciosa aguda grave, que por su letalidad y costos en atención de salud genera un alto impacto en Salud Pública. Los agentes causales más frecuentes son Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae y Listeria monocytogenes, pero poco conocemos de nuestra realidad local. MATERIALES Y MÉTODOS: estudio descriptivo, con revisión de base de datos del laboratorio de microbiología del Hospital Carlos Van Buren, obteniendo datos de los cultivos de líquido céfalo raquídeo de pacientes mayores de 15 años entre marzo de 2013 y noviembre de 2016. RESULTADOS: 128 casos de meningitis bacteriana aguda, de los cuales 17 fueron por los microorganismos objetivos del estudio, siendo el más frecuente S. pneumoniae, clínicamente un 58% se presentó sin signos meníngeos. A 30 días del diagnóstico un 35% había fallecido, la mitad de ellos inició el tratamiento antibiótico pasadas las 24 horas desde su ingreso al hospital. En el 46% la tinción gram no evidenció bacterias. DISCUSIÓN: los microorganismos clásicamente descritos como agentes causales parecen no explicar la totalidad de los cuadros de meningitis bacteriana aguda en la población adulta estudiada, la ausencia de signos meníngeos no permite descartar la sospecha diagnóstica. La mitad de los pacientes fallecidos inició el tratamiento antibiótico pasadas las primeras 24 horas.

INTRODUCTION: bacterial meningitis is a serious acute infectious disease whose lethality and elevated health costs have a serious impact on public health. The most frequent causes are Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenza and Listeria monocytogenes, but we know little of the local situation. MATERIALS AND METHODS: A descriptive study reviewing Carlos van Buren Hospital´s microbiology laboratory data base, and obtaining the details of cerebrospinal fluid cultures of patients over the age of 15 between March 2013 and November 2016. RESULTS: 128 cases of acute bacterial meningitis of which 17% were caused by the micro-organisms of study, the most frequent being Streptococcus pneumoniae. 58% of patients had no meningeal signs. At 30 days from diagnosis 35% had died, half of these having started antibiotic treatment over 24 hours after admission. 46% of the Gram stains showed no bacteria. DISCUSSION: the classical infectious agents do not appear to account for the totality of acute bacterial meningitis in the population studied. The absence of meningeal signs should not rule out the diagnosis. Half of the patients who died started antibiotic treatment after the first 24 hours.

Invasive Haemophilus influenzae disease in the vaccine era in Rio de Janeiro, Brazil

BACKGROUND Haemophilus influenzae (Hi) serotype b (Hib) conjugate vaccine was incorporated into the infant immunisation schedule in Brazil in 1999, where Hib was one of the major etiologic sources of community-acquired bacterial meningitis. OBJECTIVES The purpose of this study is to describe the molecular epidemiology of invasive Hi disease in Rio de Janeiro state, Brazil, before and after vaccine introduction. METHODS Surveillance data from 1986 to 2014 were analysed. Hi isolates recovered from cerebrospinal fluid (CSF) or blood from 1993 to 2014 were serotyped by slide agglutination, genotyped by multilocus sequence typing (MLST), and the capsule type evaluation, differentiation of serologically non-typeable isolates, and characterisation of the capsule (cap) locus was done by polymerase chain reaction. Antimicrobial susceptibility testing was performed using E-test. FINDINGS From 1986 to 1999 and from 2000 to 2014, 2580 and 197 (42% without serotype information) confirmed cases were reported, respectively. The case fatality rate was 17% and did not correlate with the strain. Hib and b- variant isolates belonged to ST-6, whereas serotype a isolates belonged to the ST-23 clonal complex. Serotype a appeared to emerge during the 2000s. Non-encapsulated isolates were non-clonal and distinct from the encapsulated isolates. Ampicillin-resistant isolates were either of serotype b or were non-encapsulated, and all of them were β-lactamase-positive but amoxicillin-clavulanic acid susceptible. MAIN CONCLUSIONS Although Hi meningitis became a relatively rare disease in Rio de Janeiro after the introduction of the Hib conjugate vaccine, the isolates recovered from patients have become more diverse. These results indicate the need to implement an enhanced surveillance system to continue monitoring the impact of the Hib conjugate vaccine.

Altered indirect hemagglutination method for easy serotyping of Haemophilus parasuis / Método de Hemaglutinação Indireta alterado para facilitar a sorotipificação de Haemophilus parasuis

Glässer's disease is an emergent bacterial disease that affects swine husbandries worldwide causing important economic losses. The aetiological agent, Haemophilus parasuis, is currently divided in fifteen serovars but an increasing number of non-typeable serovars have been reported. Indirect hemagglutination (IHA) is indicated as a serotyping method for H. parasuis. In the present study, we describe an additional step that aims to work around a possible obstacle in the original protocol that may compromise the outcome of this assay. We observed that the choice of anticoagulant for blood collection influences and/or impairs spontaneous adsorption of H. parasuis antigens on sheep red blood cells (SRBCs). However, regardless of the anticoagulant used, chemical treatment of SRBCs with tannic acid induces a stable antigen adsorption (sensitization step). The addition of 1% BSA to SRBCs washing buffer and to antisera dilution augments IHA specificity. Tannic acid treated SRBCs combined with thermo-resistant H. parasuis antigens increases the assay resolution. Thus, our results demonstrate an improvement in the technique of H. parasuis serotyping that will prove valuable to understand Glässer's disease epidemiology and to better characterize serovars involved in outbreaks.(AU)

A Doença de Glässer é uma doença bacteriana emergente que afeta a produção de suínos em todo o mundo e causa importantes perdas econômicas. O agente etiológico, Haemophilus parasuis, é atualmente dividido em quinze sorovares; no entanto, um número crescente de cepas não tipificáveis tem sido relatado. O teste de hemaglutinação indireta (IHA) tem sido utilizado para a sorotipificação de H. parasuis. Neste estudo, descrevemos uma alteração no protocolo original de IHA e que supera uma limitação específica que pode comprometer o uso geral deste ensaio. Descobrimos que o tipo de anticoagulante utilizado para coletar os eritrócitos ovinos (SRBCs) pode comprometer a adsorção espontânea dos antígenos do H. parasuis. Por outro lado, o tratamento químico dos SRBCs com ácido tânico promove uma adsorção antigênica estável (passo de sensibilização) e independente do anticoagulante utilizado. O uso de 1% de BSA durante as lavagens dos SRBCs e na diluição dos antissoros incrementa a especificidade da IHA e, a combinação dos SRBCs tratados quimicamente com antígenos de H. parasuis termo-resistentes aumentam a resolução da IHA. Nossos resultados destacam uma melhoria na principal técnica de sorotipificação de H. parasuis, que auxiliará diretamente no entendimento da epidemiologia da Doença de Glässer e na caracterização dos sorovares envolvidos em surtos da doença.(AU)

Multicenter prospective epidemiological studies on Haemophilus influenzae infection among hospitalized children with lower respiratory tract infections / 中华儿科杂志

<p><b>OBJECTIVE</b>To understand epidemiological characteristics of Haemophilus influenzae (Hi) infection in hospitalized children with lower respiratory tract infection (LRTI) in west Sichuan China.</p><p><b>METHOD</b>The multicenter prospective cross-sectional design was used; four hospitals in west Sichuan China were chosen as research field, sputum bacterial culture was done and biological typing, PCR identification and drug sensitivity test of Hi epidemic strains were carried out among 0-17y hospitalized patients with LRTI in four hospitals located in west Sichuan China.</p><p><b>RESULT</b>Totally 5 748 cases with LRTI in four hospitals were investigated in west Sichuan from Nov. 2013 to April 2014 and the rate of sputum culture was 46.96% (2,699/5 748). The total pathogenic bacteria positive rate of sputum culture was 43.53% (1,175/2 699), and 279 Haemophilus influenzae (Hi) strain in 272 cases were isolated, the Hi positive rate was 10.08% (272/2 699). All the strains (100%) were non-typeable Haemophilus influenzae (NTHi ) indentified by PCR. The main biotype of 279 strains was type Ⅰ with 39.07% (109/279) and type Ⅳ with 50.90% (142/279) ; 272 cases were enrolled in this survey, 12.50% (34/272) had broncheolitis, the rest of lower respiratory infection was 87.50 % (238/272), and 2.57% (7/272) was neonatal pneumonia, 2.21%(6/272)was pneumonia complicated with sepsis; in four hospitals the overall positive rate of Hi in inpatients with lower respiratory infection was 10.21%, 28.96%, 4.80%, 10.21% (χ(2) = 112.561, P = 0.000) and the positive rate of Hi inpatients with broncheolitis was 11.92%, 20.93%, 4.76%, and 66.67% (Fisher exact probability P = 0.001), with the rest lower respiratory infection was 9.96%, 30.90%, 4.81%, 9.85% (χ(2) =108.876, P = 0.000); 2.87% (8/279) bacterial strains of β-lactamase-nonproducing-ampicillin-intermediary (BLNAI) distributed in four hospitals, and 1.79% (5/279) bacterial strains of β-lactamase-nonproducing-ampicillin-resistant (BLNAR), 0.72% (2/279) bacterial strains of β-lactamase-positive amoxicillin-clavulanate-resistance (BLPACR) were found in two hospitals respectively.</p><p><b>CONCLUSION</b>All the Hi isolated from sputum were non-typeable among 0-17y inpatients with LRTI and the main biotype were type Ⅰ and type Ⅳ in west Sichuan China. Much attention should be paid to BLNAR and BLPACR strains found in the west Sichuan region.</p>

Evaluation of Haemophilus influenzae type b carrier status among children 10 years after the introduction of Hib vaccine in Brazil

The aim of the present study was to assess the prevalence of Haemophilus influenzaetype b (Hib) nasopharyngeal (NP) colonisation among healthy children where Hib vaccination using a 3p+0 dosing schedule has been routinely administered for 10 years with sustained coverage (> 90%). NP swabs were collected from 2,558 children who had received the Hib vaccine, of whom 1,379 were 12-< 24 months (m) old and 1,179 were 48-< 60 m old. Hi strains were identified by molecular methods. Hi carriage prevalence was 45.1% (1,153/2,558) and the prevalence in the 12-< 24 m and 48-< 60 m age groups were 37.5% (517/1,379) and 53.9% (636/1,179), respectively. Hib was identified in 0.6% (16/2,558) of all children in the study, being 0.8% (11/1,379) and 0.4% (5/1,179) among the 12-< 24 m and 48-< 60 m age groups, respectively. The nonencapsulate Hi colonisation was 43% (n = 1,099) and was significantly more frequent at 48-< 60 m of age (51.6%, n = 608) compared with that at 12-< 24 m of age (35.6%, n = 491). The overall resistance rates to ampicillin and chloramphenicol were 16.5% and 3.7%, respectively; the co-resistance was detected in 2.6%. Our findings showed that the Hib carrier rate in healthy children under five years was very low after 10 years of the introduction of the Hib vaccine.

Clinical Analysis of 35 Inpatients with Diffuse Panbronchiolitis in Peking Union Medical College Hospital / 中国医学科学院学报

<p><b>OBJECTIVE</b>To summarize the clinical features of diffuse panbronchiolitis (DPB).</p><p><b>METHOD</b>We retrospectively analyzed the clinical data of 35 patients who had been admitted to Peking Union Medical College Hospital from December 1996 to July 2014 due to DPB,which was confirmed basing on the diagnostic criteria proposed in 1998 by a working group of the Ministry of Health and Welfare of Japan or histopathological examination.</p><p><b>RESULTS</b>The average age of these 35 patients (20 men and 15 women,with a sex ratio of 1.33 to 1) was (42.2<15.6) years,mainly distributed in the 40-49 age group. The average clinical history was (8.4<8.5) years. The main symptoms and signs of DPB included chronic cough (n=35,100%),copious purulent sputum production (n=31,88.6%),exertional dyspnoea (n=24,68.6%),end-inspiratory crackles (n=28,80.0%). Also,26 patients (74.3%) had a history of sinusitis. Cold agglutination test in 15 out of 15 patients were negative. Pseudomonas aeruginosa and Haemophilus influenza were isolated from 22 patients (73.3%,22/30),and 26 patients (83.9%,26/31) had hypoxemia. The mean values of forced expiratory volume in the first second/forced vital capacity,residua volume/total lung volume,maximum forced expiratory volume of 50% lung volume,and maximum forced expiratory volume of 25% lung volume were 60.5%,53.8%,25.9%,and 31.2%,respectively. The most common CT findings from this cohort of patients were bronchiectasis and bronchiolitis,with nodular shadows distributed in a centrilobular pattern. Finally,29 patients were misdiagnosed as other conditions such as pulmonary infection and bronchiectasis.</p><p><b>CONCLUSIONS</b>DPB in Chinese populations have different presentations compared to that Japanese populations:for instance,the serum cold agglutination test always shows negative results,which is often inconsistent with the pathogens in sputum. DPB usually is misdiagnosed. Clinicians should take DPB into consideration when patients had pulmonary infection and sinusitis.</p>