Structural stability of Hib capsular polysaccharide in vaccine manufacturing / 中国生物制品学杂志

@#Objective To evaluate the stability of polyribosylribitol phosphate(PRP),the basic structure of capsular polysaccharide of Haemophilus influenzae type b(Hib),in the preparation of Hib conjugate vaccine.Methods The structures of the prepared Hib polysaccharides,polysaccharide derivatives and protein-conjugated polysaccharides were analyzed by nuclear magnetic resonance spectroscopy(NMR).Results The detection results of the prepared Hib polysaccharides,polysaccharide derivatives and protein-conjugated polysaccharides all met the requirements of relevant standards of Chinese Pharmacopoeia(VolumeⅢ,2020 edition),and the NMR spectra showed no significant change.Conclusion The basic structure PRP of the main carbohydrate antigen of Hib conjugate vaccine had no change during the vaccine manufacturing.

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.

Laboratory surveillance of invasive Haemophilus influenzae disease in Argentina, 2011-2019

Abstract The incorporation of Haemophilus influenzae type b (Hib) vaccine into the Argentine National Immunization Program in 1998 resulted in a dramatic decrease in the incidence of invasive disease due to this serotype. We assessed 1405 H. influenzae (Hi) isolates causing invasive infections referred to the National Reference Laboratory between 2011 and 2019. Non-encapsulated Hi were the most common strains (44.5%), followed by types b (41.1%) and a (10.0%). Significant increase in the proportion of type b was observed, from 31.2% in 2011, to 50% in 2015, correlating with the peak incidence rate, later decreasing to 33.6% by 2019. We compared the genetic relationship between clones circulating during the period of increased Hib incidence (2011-2015) and those of the prevaccination-transition period (1997-1998). Four pulsotypes predominated in both periods, G, M, P and K, G being the most common. Multilocus sequence typing revealed that the 4 pulsotypes belonged to ST6, or one of its simple or double locus variants. Isolates from fully vaccinated individuals did not differ from those of the rest of the population studied. After ruling out aspects associated with emergence of specific clones, we concluded that factors such as low booster coverage rates, delayed vaccination schedules and use of different vaccines may have contributed to the reemergence of Hib infections.

Resumen La introducción de la vacuna contra Haemophilus influenzae tipo b (Hib) en el Programa Nacional de Inmunización de Argentina en 1998 produjo una drástica disminución de la incidencia de enfermedad invasiva causada por este serotipo. En el Laboratorio Nacional de Referencia se estudiaron 1405 aislamientos de H. influenzae causantes de enfermedad invasiva recibidos en el período 2011-2019. H. influenzae no capsulado fue el más frecuente (44,5%), seguido por los tipos b (41,1%) y a (10,0%). Se observó un aumento significativo de la proporción del tipo b, de 31,2% en 2011 a 50% en 2015, que se correlacionó con un pico de incidencia en ese mismo año. Hacia 2019, descendió a 33,6%. Con el objetivo de evaluar los clones circulantes durante el incremento de la proporción de Hib y comparar con el período prevacunal-transición, se determinó la relación genética de una selección de aislamientos de los períodos 1997-1998 y 2011-2015. El análisis por PFGE mostró 4 pulsotipos predominantes en los 2 períodos, G, M, P y K, y el pulsotipo G fue mayoritario en ambos períodos. Por MLST se demostró que los 4 pulsotipos pertenecieron al ST6 o sus variantes (simple o doble locus). Entre los aislamientos de pacientes con vacunación completa no se hallaron clones diferentes respecto del resto de la población. Se postula que las coberturas de vacunación no satisfactorias en las dosis de refuerzo, los esquemas atrasados y el uso de diferentes vacunas pudieron haber contribuido a la reemergencia de Hib.

Orquiepididimitis por Haemophilus parainfluenzae: caso clínico y revisión de la literatura / Orchiepididymitis due to Haemophilus parainfluenzae: case report and review of the literature / Orquiepididimite por Haemophilus parainfluenzae: caso clínico e revisão da literatura

Resumen Haemophilus parainfluenzae forma parte de la microbiota normal de la cavidad oral y del tracto respiratorio superior. Es un reconocido agente causal de endocarditis y, con menor frecuencia, de enfermedades como neumonía, sepsis, osteomielitis, celulitis, meningitis y gastroenteritis aguda. Aquí se presenta un caso de orquiepididimitis en un joven adulto donde H. parainfluenzae, confirmado por espectrometría de masas (MALDI-TOF MS), fue el único patógeno detectado. Este caso contribuye a valorar el rol de H. parainfluenzae como patógeno humano, aislado a partir de sitios diferentes del torrente sanguíneo y las vías respiratorias.

Abstract Haemophilus parainfluenzae is part of the normal microbiota of the oral cavity and the upper respiratory tract. It is a recognised causal agent of endocarditis and, less frequently, of diseases such as pneumonia, sepsis, osteomyelitis, cellulitis, meningitis, and acute gastroenteritis. A case of orchiepididymitis in a young adult is reported, where H. parainfluenzae, confirmed by mass spectrometry (MALDI-TOF MS), was the only pathogen detected. This case contributes to assess the role of H. parainfluenzae as a human pathogen, isolated from sites other than the bloodstream and the respiratory tract.

Resumo Haemophilus parainfluenzae faz parte da microbiota normal da cavidade oral e do trato respiratório superior. É um reconhecido agente causal de endocardite e, menos frequentemente, de doenças como pneumonia, sepse, osteomielite, celulite, meningite e gastroenterite aguda. Aqui é relatado um caso de orquiepididimite em um adulto jovem onde H. parainfluenzae, confirmado por espectrometria de massa (MALDI-TOF MS), foi o único patógeno detectado. Este caso contribui para avaliar o papel do H. parainfluenzae como patógeno humano, isolado de outros locais que não sejam a corrente sanguínea e o trato respiratório.

Quantitative determination of cyanide residues in polysaccharide-protein conjugate vaccines based on flow injection analysis(FIA) with amperometric detector / 中国生物制品学杂志

@#Objective To develop and verify a method for determination of cyanide residues in polysaccharide-protein conjugate vaccines based on flow injection analysis(FIA)with amperometric detector.Methods After removing macromolecular substances in the samples by ultrafiltration,the cyanide residues were determined by a 3700 automatic chemical analyzer with the injection time of 35 s,injection volume of 200 μL,pump speed of 40%,sample cycle time of 140 s,ultraviolet wavelength of 312 nm and an Ampere detector.The developed method was verified for the specificity,matrix effect,linear range,limit of detection(LOD),limit of quantitation(LOQ),accuracy,presicion and stability.The cyanide residues in the polysaccharide derivative bulk(13 batches)and conjugate bulk(21 batches)of Haemophilus influenzae type b conjugate vaccine and group A and C meningococcal conjugate vaccine produced by 5 manufacturers were determined by the developed method.Results Blank samples showed no interference to detection;The recoveries of the matrix effector solution of Haemophilus influenzae type b conjugate vaccine polysaccharide derivative and conjugate bulk,group A and C meningococcal conjugate vaccine polysaccharide derivative and conjugate bulk were 97.4%,102.4%,96.8% and 99.8% respectively,with all CV values less than 15%;In the range of 0.312 5 ～ 80 ng/mL,cyanogroup concentration showed a good linear relationship with peak height with a regression equation of y = 133.13 x + 57.556,R2= 0.999 1;The LOD was 0.2 ng/mL,and the LOQ was 0.6 ng/mL.The average recoveries of the control solution were 108.9%,106.5%,103.5% respectively with RSD value of 6.4% in the groups added with 5,10 and 20 ng/mL of cyanogroup and the CV values of precision verification were all less than 15%;The average concentrations of the control solution injected 20 times continuously were 76,38,18and 5 ng/mL with all CV values less than 15%,when cyanogroup concentration was 80,40,20 and 5 ng/mL,respectively.Cyanide residues were detected in 13 batches of derivative bulk samples,while not in 2 batches of 21 batches of polysaccharide conjugate bulk samples,and were detected in the others.Conclusion The developed method had good accuracy,precision and stability,which might be applied to the quantitative determination of cyanide residues in polysaccharideprotein conjugate vaccines.

Niveles de protección contra Haemophilus influenzae tipo b en niños, adolescentes y adultos cubanos / Levels of protection against Haemophilus influenzae type b in Cuban children, adolescents and adults

La concentración de los anticuerpos contra el polisacárido capsular polirribosilribitol fosfato del Haemophilus influenzae tipo b se considera un buen indicador serológico para evaluar protección contra la enfermedad invasiva. Existen pocos reportes que estudien la inmunidad serológica en Cuba. El objetivo general de este estudio fue determinar los niveles de protección séricos contra Haemophilus influenzae tipo b en niños, adolescentes y adultos cubanos, en una muestra de 575 individuos. Se cuantificó la concentración de IgG anti-polirribosilribitol fosfato de Haemophilus influenzae tipo b mediante un inmunoensayo enzimático estandarizado y validado en el laboratorio de inmunología del Centro Nacional de Genética Médica, La Habana, Cuba. Se determinaron las concentraciones medias geométricas de anticuerpos y los niveles de protección frente a la enfermedad invasiva por Haemophilus influenzae tipo b. La concentración media geométrica de IgG anti-polirribosilribitol fosfato fue de 1,94 μg/mL (IC95 por ciento 1,80; 2,08) y fue mayor en el grupo de 16 a 22 años. El porcentaje con protección de larga duración fue mayor para el sexo femenino que para el masculino (82,2 por ciento vs 71,4 por ciento; p=0,0339) entre los que poseían inmunidad natural. El grupo de sujetos nacidos en el periodo en que se vacunó con la vacuna conjugada cubana QUIMI-HIB® presentó concentraciones medias geométricas superiores (2,75 μg/mL, IC95 por ciento 2,00; 3,79). El 99,1 por ciento de los participantes presentó protección frente a la enfermedad invasiva por Haemophilus influenzae tipo b, el 19,8 por ciento a corto plazo y el 79,3 por ciento protección de larga duración. El inmunoensayo validado para la cuantificación de IgG anti-polirribosilribitol fosfato podría emplearse en estudios de seroprevalencia. En los sujetos estudiados, se encontró un predominio de elevadas concentraciones de IgG anti- polirribosilribitol fosfato del Haemophilus influenzae tipo b que confieren protección de larga duración(AU)

The levels of antibodies directed against the capsular polysaccharide polyribosylribitol phosphate of Haemophilus influenzae type b are considered a good serological indicator to assess the immunity against invasive disease. In Cuba, there are few reports that study serological immunity. The general objective was to determine serum protection levels against Haemophilus influenzae type b in Cuban children, adolescents and adults, in a sample of 575 Cuban individuals. The concentration of IgG against Haemophilus influenzae type b was quantified by means of an indirect ELISA standardized and validated in the immunology laboratory of the National Center of Medical Genetics, Havana, Cuba. The geometric mean concentration of IgG anti- polyribosylribitol phosphate and the levels of protection against invasive Haemophilus influenzae type b disease were determined. The geometric mean concentration of IgG anti- polyribosylribitol phosphate was 1.94 μg/mL (95percentCI 1.80;2.08) and the group from 16 to 22 years old presented the highest. Among those with natural immunity, the percentage with long-term protection was higher for females vs. males (82.2percent vs. 71.4percent; p=0.0339). The group of subjects born in the period in which they were vaccinated with the Cuban conjugate vaccine QUIMI-HIB® presented higher geometric mean concentration (2.75 μg/mL, CI95percent 2.00; 3.79). The 99.1percent of the participants had protection against invasive Haemophilus influenzae type b disease, 19.8percent short-term and 79.3percent long-term protection. The ELISA for the quantification of anti- Haemophilus influenzae type b IgG antibodies, developed and validated, could be used in seroprevalence studies. In the subjects studied, there was a predominance of high IgG anti- Haemophilus influenzae type b polyribosylribitol phosphate concentration values that confer long-term protection(AU)

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)

COVID-19 and influenza: implications in the public health context / COVID-19 e influenza: implicações no contexto da saúde pública

The COVID-19 pandemic caused by the novel coronavirus SARS-CoV-2 continues to have a major impact on health and social systems around the world. The COVID-19 and influenza manifest themselves in a similar way, causing respiratory diseases that can present asymptomatically, as well as from the cold to severe respiratory problems until death. The form of transmission is similar, through contact with droplets or particles of saliva and secretions, which implies preventive actions that involve the same hygiene measures, use of masks and the need to cough using the elbow or disposable tissues. This characterizes a syndemic. It becomes necessary to monitor these diseases so that there are parameters for better decision-making on the appropriate clinical management of these respective diseases.

A pandemia de COVID-19 causada pelo novo coronavírus SARS-CoV-2 continua a ter um grande im-pacto nos sistemas de saúde e sociais em todo o mundo. torna-se necessário que haja monitoramento dessas doenças para que assim, tenha-se parâmetros para melhor tomada de decisão sobre a gestão clínica adequada sobre essas respectivas doenças. Os dois vírus se manifestam de forma semelhante, ao causarem doenças respiratórias que podem se apresentar de forma assintomáticas, assim como do resfriado a problemas respiratórios graves até a morte. A forma de transmissão são parecidas, por contato com gotículas ou partículas de saliva e secreções, o que implica nas ações de prevenção que perpassam pelas mesmas medidas higienização, uso de máscaras e a necessidade de tossir usando o cotovelo ou lenções descartáveis. Isto caracteriza uma sindemia.

In vitro antibacterial activity of pediatric Faropenem sodium against respiratory tract pathogens in children / 中华实用儿科临床杂志

Objective:To evaluate the antibacterial activity of pediatric Faropenem sodium against common pathogens isolated from children′s respiratory tract in vitro, and to provide reference for its clinical research and application. Methods:Retrospective analysis.The minimum inhibitory concentration (MIC) of Faropenem sodium, Merope-nem, Imipenem and other antibiotics was determined by the agar dilution method.A total of 156 strains of Streptococcus pneumoniae [including 32 strains of Penicillin-susceptible Streptococcus pneumoniae (PSSP), 28 strains of Penicillin-intermediate Streptococcus pneumoniae (PISP) and 96 strains of Penicillin-resistant Streptococcus pneumoniae (PRSP)], 98 strains of Haemophilus influenza, 173 strains of Klebsiella pneumoniae, and 55 strains of Moraxella catarrhali clinical isolates were used.MIC 50, MIC 90 and the accumulative inhibition of the bacteria were investigated. Results:The MIC of Faropenem sodium against all the Streptococcus pneumoniae strains ranged from 0.010-2.000 mg/L.There was no difference in the MIC distribution of Faropenem sodium against PSSP, PISP and PRSP, and the MIC 90 value was all 1.000 mg/L.Faropenem sodium inhibited all the Haemophilus influenza strains at concentrations ranging from 0.030-8.000 mg/L.There was no difference in the MIC distribution of Faropenem sodium against Haemophilus influenza with or without β-lactamase and Ampicillin resistance.The MIC 90 value was all 4.000 mg/L.Ho-wever, the MIC of Faropenem sodium against Klebsiella pneumoniae ranged from 0.250 to above 32.000 mg/L, and both MIC 50 and MIC 90 were greater than 32.000 mg/L.Faropenem sodium inhibited all the Moraxella catarrhalis strains at concentrations ranging from 0.030-2.000 mg/L, with MIC 50 being 0.500 mg/L and MIC 90 being 1.000 mg/L. Conclusions:Antimicrobial susceptibility testing results in vitro demonstrate that pediatric Faropenem sodium has satisfactory antibacterial activities against Streptococcus pneumoniae, Haemophilus influenza, and Moraxella catarrhalis, but comparatively weak antibacterial activities against Klebsiella pneumoniae.

Common bacteria in oropharynx of children with Mycoplasma pneumoniae pneumonia and its clinical significance / 中华实用儿科临床杂志

Objective:To investigate the common bacteria in the oropharynx of children with Mycoplasma pneumoniae pneumonia (MPP) and its clinical significance.Methods:A total of 134 children with MPP who were hospitalized in the Department of Pediatric Respiratory, Shengjing Hospital of China Medical University from December 2016 to June 2017 were selected as the research subjects, and 42 healthy children in the same hospital were selected retrospectively as the healthy control group during the same period.Fluorescent quantitative polymerase chain reaction Taqman probe was used to detect common oropharyngeal bacteria[ Streptococcus pneumoniae(SP), Moraxella catarrhalis(CTA), Haemophilus influenza(HI)] for the enrolled children.Firstly, the bacterial detection rate of MPP children and healthy children was compared.Then, according to age(<1 years old, 1-<3 years old, 3-<6 years old and 6-14 years old), bacterial detection[Mycoplasma pneumoniae(MP), MP+ bacteria]and bacterial species(MP+ SP, MP+ CTA, MP+ HI), 134 children with MPP were divided into groups to compare.Moreover, the relevant clinical datas were retrospectively analyzed by rank sum test and chi- square test. Results:Among 134 children with MPP, 79 (58.96%) children were detected bacteria, and 17 (40.48%) children were detected bacteria among 42 healthy children, with statistically significant differences( χ2=4.404, P<0.05). Compared with the MP group, the level of white blood cell (WBC)[8.5(6.7, 12.0)×10 9/L vs.7.8(5.8, 9.3)×10 9/L, Z=-2.232], C reactive protein(CRP)[19.2(7.2, 35.0) mg/L vs.8.4(3.4, 24.6) mg/L, Z=-2.810], lactate dehydrogenase(LDH)[286(244, 365) U/L vs.250(210, 302) U/L, Z=-2.474] and the incidence of lobar pneumonia[40.51%(32/79 cases) vs.18.18%(10/55 cases), χ2=7.510], pleural effusion[13.92%(11/79 cases) vs.3.64%(2/55 cases), χ2=3.917], refractory Mycoplasma pneumoniae pneumonia (RMPP)[34.18%(27/79 cases) vs.18.18%(10/55 cases), χ2=4.151] in MP+ bacteria group were higher; the course of fever[10(7, 12) d vs.8(6, 10) d, Z=-2.706] and duration of antibiotic use[16(13, 19) d vs.12(9, 16) d, Z=-3.747] in MP+ bacteria group were longer (all P<0.05). The level of WBC in MP+ SP group[12.20(7.80, 17.30)×10 9/L] was higher than that in MP+ HI group [6.75(5.37, 9.44)×10 9/L], and the differences were statistically significant( Z=11.574, P<0.05), and the incidence of lobar pneumonia in MP+ SP group [56.67%(17/30 cases)]was higher than that in MP+ CTA group [0(0/3 cases)]and MP+ HI group[18.75%(3/16 cases)], and the differences were statistically significant( χ2=9.770, P<0.05). Conclusions:Bacterial colonization or infection is more likely to occur in the oropharynx of children with MPP.When WBC, CRP, and LDH are significantly increased and the image shows a large consolidation or pleural effusion, it may indicate mixed bacterial infection, longer course of fever and higher incidence of RMPP, and the common mixed bacteria is SP.

A case report of intracranial infection caused by Aggregatibacter aphrophilus / 中华神经科杂志

Aggregatibacter aphrophilus is a member of the normal flora of the human oral cavity and pharynx, a Gram-negative fastidous bacteria, belonging to agglomerates, which is a normal mixed oropharyngeal flora in humans, most commonly colonized on the surface of oral mucosa. This bacterial infection is rare in clinical practice, and it is difficult to culture and identify the bacteria, which is easy to be missed. A patient with intracranial infection was admitted to Huaihe Hospital, who showed fever and headache as the main clinical manifestations, and Aggregatibacter aphrophilus was detected by the metagenomic next-generation sequencing of cerebrospinal fluid. The patient′s symptoms were significantly improved after anti-infection treatment.

Analysis of serotype distribution and antibiotic resistance of Haemophilus influenzae in hospitalized children / 中华实用儿科临床杂志

Objective:To explore the serotype, antibiotic resistance and β-lactamase gene of Haemophilus influenzae strains isolated from hospitalized children, thus providing reference for clinical diagnosis and treatment. Methods:A total of 148 Haemophilus influenzae strains collected from January 2016 to December 2018 in hospitalized children of Children′s Hospital, Capital Institute of Pediatrics were retrospectively analyzed.The serotype and genotype of Haemophilus influenzae strains were examined by slide agglutination test and PCR, respectively.The sensitivity of isolates to Ampicillin and other antimicrobials was detected by the E-test and disk diffusion methods.The β-lactamase phenotype was tested by nitrocefin disk method.The carrying of β-lactamase gene TEM-1 and ROB-1 were detected by PCR.The drug resistance rate was compared by χ2 test. Results:All the 148 strains were nontypeable Haemophilus influenzae (NTHi), and capsular gene was not amplified.The rate of resistance to Ampicillin, Ampicillin/Sulbactam, Cefuroxime, and Azithromycin were 68.9%(102/148 strains), 40.5%(60/148 strains), 53.4%(79/148 strains) and 56.1%(83/148 strains), respectively.The Haemophilus influenzae isolates showed the highest resistance rate to Trimethoprim-sulfamethoxazole, which was up to 91.9%(136/148 strains). The sensitive rate of isolates to Ceftriaxone, Meropenem and Levofloxacin were all 100.0%(148/148 strains). The prevalence of β-lactamase was 64.8%(96/148 strains) in Haemophilus influenzae and the genotype was TEM-1.The drug resistance rates of β-lactamase positive strains to Ampicillin, Ampicillin/Sulbactam and Azithromycin were significantly higher than those of other strains( χ2=123.222, 27.973, 70.273, all P<0.01). Conclusions:The most prevalent serotype of Haemophilus influenzae is NTHi in children. Haemophilus influenzae carried TEM-1 gene had a high positive rate of β-lactamase production, which was the main mechanism of drug resistance to Ampicillin.Ceftriaxone and Meropenem were the most active agents against Haemophilus influenzae.

Whole-genome analysis of haemophilus influenzae invasive strains isolated from Campinas state University hospital. An epidemiological approach 2012 - 2019 and ancestor strains

Abstract Thirteen Haemophylus influenzae invasive strains isolated from patients at Clinical Hospital of State University of Campinas, from May 2013 through August 2019, was submitted to Illumina genome sequencing HiSeq platform. Further in silico analysis of serogroup and Multi Locus Sequence Typing (MLST) from whole DNA sequencing had demonstrated the actual clonal distribution in the Campinas Metropolitan region. Thus, results showed the existence of a new ST Haemophilus influenzae found in the Brazilian territory and an increase of strains belonging to serogroup a (three strains also belonging to ST23). In conclusion, we observed an increase of non-typable H. influenzae (NTHi) and a strain involved in invasive diseases in the Campinas - São Paulo region after frequent detection of those serotypes and genotypes in other Brazilian regions.

Sentinel surveillance for bacterial pneumonia and meningitis in children under the age of 5 in a tertiary pediatric hospital in Colombia - 2016 / Vigilancia centinela de neumonías y meningitis bacterianas en menores de 5 años en un hospital pediátrico de tercer nivel en Colombia - 2016

Abstract | Introduction: Bacterial pneumonia and meningitis are vaccine-preventable diseases. Sentinel surveillance provides relevant information about their behavior. Objective: To present the data from sentinel surveillance carried out at the Fundación HOMI, Fundación Hospital Pediátrico La Misericordia in 2016. Materials and methods: We conducted a descriptive study from January 1 to December 31, 2016, on the daily surveillance of patients under 5 years of age diagnosed with pneumonia or bacterial meningitis according to PAHO's definitions. We identified the microorganisms using the automated VITEKTM 2 system. Bacterial isolates were sent to the Microbiology Group at the Colombian Instituto Nacional de Salud for confirmation, serotyping, phenotypic, and genotypic characterization. Antimicrobial susceptibility profiles were established. Results: From 1,343 suspected cases of bacterial pneumonia, 654 (48.7%) were probable, 84% had complete Hib vaccination schedules, and 87% had complete pneumococcal vaccination schedules for age. Blood culture was taken in 619 (94.6%) and 41 (6.6%) were positive while S. pneumoniae was isolated in 17 (41%) of them. The most frequent serotype was 19A in five cases (29.4%), and four 19A serotypes were associated with the reference isolate ST320. The incidence rate of probable bacterial pneumonia was 7.3 cases/100 hospitalized patients, and lethality was 2.1%. As for bacterial meningitis, 22 suspected cases were reported, 12 (54%) were probable, four (33%) were confirmed: two by Escherichia coli and two by group C N. meningitidis. The incidence of probable bacterial meningitis was 0.14 cases/100 hospitalized patients. Conclusion: Streptococcus pneumoniae serotypes 19A and 3 were the most frequent cause of pneumonia. Spn19A is related to the multi-resistant clone ST320. Strengthening and continuing this strategy will allow understanding the impact of vaccination.

Resumen | Introducción. La neumonía y la meningitis bacterianas son enfermedades inmunoprevenibles; la vigilancia centinela aporta información relevante acerca de su comportamiento. Objetivo. Presentar los resultados de la vigilancia centinela de neumonía y meningitis llevada a cabo en la HOMI, Fundación Hospital Pediátrico La Misericordia. Materiales y métodos. Se hizo un estudio descriptivo entre el 1 de enero y el 31 diciembre del 2016, de la vigilancia diaria de pacientes menores de 5 años con diagnóstico de neumonía o meningitis bacteriana, según las definiciones de la Organización Panamericana de la Salud (OPS). Los microorganismos fueron identificados usando el sistema automatizado VITEK TM2. Los aislamientos se enviaron al grupo de microbiología del Instituto Nacional de Salud para confirmación, serotipificación, y caracterización genotípica y fenotípica. Asimismo, se establecieron los perfiles de sensibilidad antimicrobiana. Resultados. De 1.343 casos sospechosos de neumonía bacteriana, 654 (48,7 %) fueron probables, el 84 % tenía el esquema de vacunación completo para la edad contra Haemophilus influenzae de tipo b, y el 87 %, contra neumococo. En 619 (94,6 %) pacientes se hizo hemocultivo y 41 (6,6 %) fueron positivos. S. pneumoniae se aisló en 17 (41 %) casos. El serotipo más frecuente fue el 19A, en cinco pacientes (29,4 %), en tanto que cuatro aislamientos de spn19A fueron relacionados con el clon ST320. La tasa de incidencia de neumonía bacteriana probable fue de 7,3 casos/100 pacientes hospitalizados. La letalidad fue de 2,1 %. Hubo 22 casos sospechosos de meningitis bacteriana, 12 (54 %) probables, y cuatro (33 %) confirmados: dos por Escherichia coli y dos por Neisseria meningitidis del grupo C. La incidencia de meningitis bacteriana probable fue de 0,14/100 pacientes hospitalizados. Conclusión. Los serotipos 19A y 3 de S. pneumoniae fueron la causa más frecuente de neumonía. El Spn19A se relacionó con el clon ST320 mulitirresistente. El fortalecimiento continuo de la vigilancia centinela permitirá entender el impacto de la vacunación.

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)

Pediatric bacterial meningitis and meningococcal disease profile in a Brazilian General Hospital

Abstract Objective To evaluate the clinical and epidemiological profile of bacterial meningitis and meningococcal disease in pediatric patients admitted to a Brazilian Secondary Public Hospital. Methods A descriptive observational study was conducted. Microbiologically proven bacterial meningitis or meningococcal disease diagnosed from 2008 to 2018 were included. Results A total of 90 patients were diagnosed with proven bacterial meningitis. There were 64 confirmed cases of meningococcal disease. The prevalence was higher in boys (n=38), median age 30 months (1-185). The main clinical manifestations were: meningococcal meningitis (n=27), meningococcemia without meningitis (n=14), association of meningococcemia with meningitis (n=13), and fever without a known source in infants (n=7).Admissions to intensive care unit were necessary for 45 patients. Three deaths were notified. Serogroup C was the most prevalent (n=32) followed by serogroup B (n=12).Pneumococcal meningitis was identified in 21 cases; out of the total, 10 were younger than two years. The identified serotypes were: 18C, 6B, 15A, 28, 7F, 12F, 15C, 19A and 14. Pneumococcal conjugate 10-valent vaccine covered four of the nine identified serotypes.Haemophilus influenzae meningitis serotype IIa was identified in three patients, median age 4 months (4-7). All of them needed intensive care. No deaths were notified. Conclusion Morbidity and mortality rates from bacterial meningitis and meningococcal disease remain high, requiring hospitalization and leading to sequelae. Our study observed a reduced incidence of bacterial disease over the last decade, possibly reflecting the impact of vaccination.

Impacto do uso da reação em cadeia da polimerase para o diagnóstico de meningite bacteriana em substituição a contraimunoeletroforese / Impact of using polymerase chain reaction for the diagnosis of bacterial meningitis in substitution for counterimmunoelectrophoresis

Introdução: a meningite bacteriana é um grave problema de Saúde Pública mundial, tendo como principais agentes: Neisseria meningitidis, Streptococcus pneumoniae e Haemophilus influenzae. A metodologia de diagnóstico empregada no Instituto Adolfo Lutz ­ Centro de Laboratório Regional Santo André até o ano de 2011 era a contraimunoeletroforese (CIE), depois foi substituída pela reação em cadeia da polimerase em tempo real (qPCR), que apresenta maior sensibilidade. Objetivo: este trabalho objetivou comparar ambas as metodologias no período de 2009 a 2018, para avaliação do impacto da introdução da qPCR no diagnóstico das meningites bacterianas nos 7 municípios da região do ABC do Estado de São Paulo. Metodologia: foram avaliadas a quantidade total de exames realizados, a média mensal, a positividade no período, os municípios requisitantes e a prevalência das bactérias causadoras de meningite, no período de abril/2009 até dezembro/2018. Resultados: Foram 377 exames de CIE e 1305 de qPCR, com média anual de 230 exames em 2010-2013 e 130 exames em 2014-2018. Observou-se aumento da positividade entre as técnicas, 17,8% para CIE e 33,8% para qPCR. N. meningitidis foi responsável pela maioria dos casos entre 2011 e 2013, cerca de 61% dos casos positivos, enquanto que entre 2014 e 2018 foi S. pneumoniae, cerca de 53%. Conclusão: os resultados indicaram que a qPCR foi mais eficiente em detectar os agentes causadores de meningite bacteriana na região do que a técnica de CIE. Por fim, este trabalho suporta a implantação da metodologia de qPCR para diagnóstico de meningite em substituição de técnicas menos sensíveis.

Introduction: bacterial meningitis is still a serious worldwide public health problem, and the main etiological agents are: Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae. The diagnostic methodology employed at the Adolfo Lutz Institute ­ Santo André Regional Laboratory Center until 2011 was the ounterimmunoelectrophoresis (CIE), then it was replaced by the real-time polymerase chain reaction (qPCR), which is more sensitivity. Objective: this study aimed to compare both methodologies from 2009 to 2018 to evaluate the impact of the introduction of qPCR in the diagnosis of bacterial meningitis in the 7 cities of the ABC region of São Paulo State. Methodology: the total number of tests performed, the month average, the positivity in the period, the requesting cities and the prevalence of bacteria causing meningitis were evaluated from April/2009 to December/2018. Results: there were 377 CIE exams and 1305 qPCR exams, with an annual average of 230 exams in 2010-2013 and 130 exams in 2014-2018. There was an increase in positivity between the performed techniques, 17.8% for CIE and 33.8% for qPCR. N. meningitidis accounted for most cases of bacterial meningitis between 2011 and 2013, about 61% of positive cases, whereas between 2014 and 2018 it was S. pneumoniae, with about 53%. Conclusion: the results indicated that qPCR was more efficient in detecting the agents that cause bacterial meningitis in the region than the CIE technique. Finally, this work supports the implementation of qPCR methodology for diagnosis of meningitis in replacement of less sensitive techniques.

Predicting bacteria causing acute bacterial rhinosinusitis by clinical features / Predição de bactérias causadoras de rinossinusite bacteriana aguda por características clínicas

Abstract Introduction: Clinicians rely on clinical presentations to select therapeutic agents for acute bacterial rhinosinusitis. Streptococcus pneumoniae and Haemophilus influenzae are common in acute bacterial rhinosinusitis. Drug resistant Streptococcus pneumoniae and Haemophilus influenzae require different antibiotics. Objective: This study aimed to evaluate the associations between clinical features of acute bacterial rhinosinusitis and pathogenic bacteria. Methods: Sixty-four patients with acute bacterial rhinosinusitis were enrolled. Clinical features including nasal obstruction, discolored discharge, facial pain, smell disturbance, fever and laboratory findings of patients with acute bacterial rhinosinusitis were collected. The bacterial cultures of endoscopic middle meatal swabs were used as a reference. Results: Serum C-reactive protein level elevation correlated with the bacterial species (p = 0.03), by which was increased in 80.0% of Haemophilus influenzae rhinosinusitis and 57.1% of Streptococcus pneumoniae rhinosinusitis. The elevated C-reactive protein was the significant predictor for Haemophilus influenzae rhinosinusitis with the Odds Ratio of 18.06 (95% CI 2.36-138.20). The sensitivity of serum C-reactive protein level elevation for diagnosing Haemophilus influenzae rhinosinusitis was 0.80 (95% CI 0.49-0.94). Conclusion: Elevation of serum C-reactive protein level was associated with and predicted acute bacterial rhinosinusitis caused by Haemophilus influenzae.

Resumo: Introdução: Os médicos se baseiam nas características clínicas para a escolha dos agentes terapêuticos para o tratamento da rinossinusite bacteriana aguda. Streptococcus pneumoniae e Haemophilus influenzae são agentes comuns na rinossinusite bacteriana aguda. Streptococcus pneumoniae e Haemophilus influenzae resistentes a antibióticos requerem medicamentos diferentes. Objetivo: Avaliar as associações entre as características clínicas da rinossinusite bacteriana aguda e bactérias patogênicas. Método: O estudo incluiu 64 pacientes com rinossinusite bacteriana aguda. Foram coletadas e registradas as características clínicas, inclusive obstrução nasal, secreção com cor alterada, dor facial, distúrbios do olfato, febre e achados laboratoriais de pacientes com rinossinusite bacteriana aguda. As culturas bacterianas obtidas por swab endoscópico do meato médio foram usadas como referência. Resultados: A elevação do nível sérico de proteína C-reativa estava correlacionada com a espécie bacteriana (p = 0,03); ela estava aumentada em 80,0% das rinossinusites por Haemophilus influenzae e em 57,1% das rinossinusites por Streptococcus pneumoniae. A proteína C-reativa elevada foi um significativo fator preditor de rinossinusite por Haemophilus influenzae, com razão de probabilidade de 18,06 (IC 95% 2,36-138,20). A sensibilidade da elevação dos níveis séricos de proteína C-reativa para o diagnóstico de rinossinusite por Haemophilus influenzae foi de 0,80 (IC 95% 0,49 ± 0,94). Conclusão: A elevação dos níveis séricos de proteína C-reativa é um preditor de rinossinusite bacteriana aguda causada por Haemophilus influenzae.

Endoftalmitis endógena como primera manifestación de meningitis por Haemophilus influenza: reporte de un caso / Endogenous endophthalmitis as the initial presentation of Haemophilus influenzae meningitis

La endoftalmitis endógena es responsable del 2-15 % de las endoftalmitis, se relaciona con una infección sistémica hasta en un 52-90% de los casos, en el contexto de infecciones sistémicas como abscesos hepáticos, neumonia, infecciones de vias urinarias, meningitis, usurios de drogas endovenosas y fungémias. Haemophilus influenzae es un agente etiológico inusual de esta patología, hay pocos casos reportados a nivel mundial y no se han reportado casos en nuestro país; las endoftalmitis por Haemophilus se caractererizan por tener un pronóstico visual sombrio, ya que pueden presentar complicaciones severas y comprometer permanentemente la visión. A continuación, se describe un caso de endoftalmitis endogéna como primera manifestación de meningitis asociada a H. Influenzae en una paciente inmunocompetente y sin factores de riesgo.

Endogenous endophthalmitis is responsible for 2-15% of endophthalmitis, it is associated with a systemic infection in up to 52-90% of cases, in the context of systemic infections such as liver abscesses, pneumonia, urinary tract infections, meningitis, intravenousdrug users and fungaemia. Haemophilus influenzae is an unusual etiologic agent of this pathology, there are few cases reported worldwide and no cases have been reported in our country. The endophthalmitis due to Haemophilus are characterized by having a somber visual prognosis, since they can present severe complications and can permanently affect vision. Mentioned hereinafter, a case of endogenous endophthalmitis is described as the first manifestation of meningitis associated with H.Influenzae in an immunocompetent patient with no risk factors.