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.

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)

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.

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.

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.

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.

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.

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.

Bacterial meningitis in patients with sickle cell anemia in Salvador, Bahia, Brazil: a report on ten cases

ABSTRACT Sickle cell anemia (SCA) is a common genetic blood disorder, affecting millions worldwide. According to current evidence, individuals with SCA have more than 300 times greater risk to develop bacterial meningitis (BM) than the general population. Herein we have described the characteristics of a series of BM cases in SCA patients in Salvador, Brazil, during 13 years of hospital-based surveillance. Data on clinical presentation, laboratory parameters and outcomes were collected retrospectively by reviewing medical records. From 1999 to 2011, ten SCA patients were identified among the 2511 cases of BM (10/2511; 0.40%). These patients were more likely to be male (90%) and to be younger (median age 8.5 years). The causative agents were Streptococcus pneumoniae (n = 5) and Haemophilus influenzae (n = 1). The most frequent pneumococcal serotypes were 23 F (2 cases), 14, 18 F, 23B (one case each). Common medical complications were stroke (n = 3); heart failure (n = 2), respiratory problems (n = 2), renal dysfunctions (n = 2) and leg ulcers (n = 1). This study highlights the importance of S. pneumoniae as a causative agent of meningitis in individuals with SCA and shows the diversity of comorbidities associated with this condition.

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.

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.

Budleja asiatica Derived Phytochemicals against Eye Disease

The secondary metabolites in the plants are used for curing various diseases. The phytochemicals present in Budleja asiaticaplant extract are traditionally used to cure Eye disease. It is caused by Haemophilus influenza, whichis a facultatively anaerobic, gram-negative pathogenic bacteria. For analysis of ligand-protein interaction, the molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that lignoceric acid can effectively deactivate Aspararte semialdehyde dehydrogenase enzyme which is important macromolecule for the regulation of amino acid biosynthesis resulting in interrupting the life cycle of the microorganism

Capsicum anum L. Derived Phytochemicals against Haemophilus influenzae Causing Bronchitis

Phytochemicals from Capsicum anum L. plant extract are traditionally used to cure bronchitis. Bronchitis is caused by Haemophilus influenzae. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that myrcetin and quercetin can effectively deactivate the Palmitoyl-CoA hydrolase enzyme thereby interrupting the life cycle of aemophilus influenza

Epiglotitis por Haemophilus influenzae tipo b en la era de la vacunación: caso clínico pediátrico / Epiglottitis due to Haemophilus influenzae type b in the vaccination era: pediatric clinical case

La epiglotitis aguda infecciosa es infrecuente en la actualidad, debido a la vacunación contra su principal agente etiológico, el Haemophilus influenzae b. Se requiere alto índice de sospecha ante el cuadro clínico de dificultad respiratoria, estridor, disfonía y fiebre. Se presenta a un niño de 2 años, previamente sano, con esquema de vacunas completas, con dificultad respiratoria aguda y estridor laríngeo, en el que, al momento de realizar la intubación, se realizó el diagnóstico de epiglotitis aguda. Con hemocultivos positivos para Haemophilus influenzae b, cumplió 13 días de tratamiento con ceftriaxona, con hemocultivos de control y cultivo de líquido cefalorraquídeo negativo.

Acute infectious epiglottitis is infrequent at present due to vaccination for its main etiologic agent, Haemophilus influenzae b (Hib). It must be taken into account when we make a differential diagnosis in a child whose clinical symptoms are respiratory distress, stridor, dysphonia and fever. We report a 2-year-old child, previously healthy, whose vaccination calendar was complete, and whose clinical presentation included respiratory distress and stridor; at the moment of the intubation the laryngoscopy showed an acute epiglottitis. Blood cultures were taken, which were positive for Hib. He was treated with ceftriaxone during 13 days, and the control blood cultures and cerebrospinal fluid were negative.

Historia de la meningitis bacteriana en Cuba: siglo XIX al XXI

Introducción: La meningitis bacteriana aún constituye un importante problema de salud mundial. En Cuba hay limitadas investigaciones con una perspectiva histórica de esta temática. Objetivo: Describir el comportamiento de la meningitis bacteriana en Cuba (siglo XIX-XXI). Material y Métodos: Estudio descriptivo (corte histórico) entre finales de 1800 y 2017, utilizando el método histórico-lógico y un análisis deductivo-inductivo de múltiples fuentes bibliográficas. Desarrollo: La primera alusión a la meningitis bacteriana en Cuba data de 1877. También se sugiere su probable importación por el ejército de ocupación norteamericano en 1899. En 1901 se aisló el meningococo de Weichselbaum del líquido cefalorraquídeo, lo que constituye, probablemente, su primera notificación en Cuba. Iniciado el siglo XX, se reportan casos aislados y brotes hasta 1976 cuando inicia la mayor epidemia de Enfermedad Meningocócica (serogrupos C y B). En 1979 se vacuna contra el C. En 1980 se implementa una vigilancia epidemiológica especial. Se desarrolla la vacuna cubana VA-MENGOC-BC® (1984) que se usa masivamente (1987) y se contribuye al control, incluyéndose en el Programa Nacional de Inmunizaciones (1991). Haemophilus Influenzae pasa a ser la principal bacteria causante de meningitis bacteriana hasta 1999 en que se aplican vacunas (Vaxem-Hib® y QuimiHib®) y se controla. Su nicho ecológico es ocupado por neumococo hasta ahora. Conclusiones: Desde fines del siglo XIX hasta la segunda mitad del XX la meningitis bacteriana en Cuba se manifestaba como casos aislados y brotes. Durante y después de una gran epidemia, se implementan estrategias preventivas efectivas, incluidas dos vacunas cubanas contra estas enfermedades, que revierten el comportamiento a endemia muy baja hasta la actualidad.

Introduction: Bacterial meningitis remains an important health problem worldwide. In Cuba, there are limited research studies about this issue from a historical perspective. Objective: To describe the behavior of this disease in Cuba (19th - 21st centuries). Material and Methods: A descriptive historical study was carried out between the ends of 1800-2017, using the historical-logical method and a deductive-inductive analysis of multiple bibliographical sources. Development: The first reference to bacterial meningitis in Cuba was made in 1877. The probable introduction of the disease by the US occupation army in 1899 is also considered. In 1901, the meningococci of Weichselbaum was isolated from cerebrospinal fluid, which was probably it first report in Cuba. At the beginning of the 20th century, isolated cases and outbreaks were reported until 1976, when the biggest and larger invasive meningococcal disease began (serogroups C, B). Vaccination against serogroup C started in 1979. In 1980, a special epidemiological surveillance was implemented. The Cuban vaccine VA-MENGOC-BC® against the disease was developed in 1984, which was massively used in 1987. The vaccine contributed to the control of the disease and was included in the National Immunization Program in 1991. Haemophilus Influenzae became the main causative bacterial agent of meningitis until 1999, when the implementation of massive vaccination (Vaxem-Hib® and QuimiHib®) controlled disease. Up to the present, its ecological niche is occupied by pneumococci. Conclusions: From the end of the 19th century to the second half of the 20th century, bacterial meningitis in Cuba behaved as isolated cases and outbreaks. Effective preventive strategies were implemented during and after a huge epidemic, including Cuban vaccines against the disease, that pass on its behavior to a very low endemic disease up to the moment.

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.

Celulitis de cara y cuello por Haemophilus influenzae tipo b. A propósito de un caso clínico / A clinical case of face and neck cellulite due to type b Haemophilus influenzae / Caso clínico de celulite de rosto e pescoço por Haemophilus influenzae tipo b

Resumen: Haemophilus influenzae tipo b es una bacteria capsulada gramnegativa de distribución mundial. Es responsable de diferentes formas clinicas de enfermedad invasiva entre las que se encuentran: meningitis, epiglotitis, bacteriemia, artritis séptica, celulitis, osteomielitis y pericarditis. Estas enfermedades son inmunoprevenibles. La celulitis facial por Haemophilus influenzae tipo b es una infección invasiva potencialmente grave que afecta predominantemente a menores de 2 años de edad, dado que es una población inmunológicamente susceptible. Existen elementos clínicos característicos, pero no patognomónicos que permiten sospechar esta infección. Se presenta el caso clínico de un lactante de 8 meses de edad, únicamente calmetizado, que acude al servicio de urgencias con una tumefacción extensa de cara que ingresa a sala de cuidados moderados con planteo clínico de celulitis. Dada la rápida extensión del proceso infeccioso, requiere traslado a unidad de cuidados intensivos. Se realizó analítica de valoración infecciosa y tomografía computada que delimitó el proceso y confirmó la existencia de una colección. Se inició antibioticoterapia empírica y drenaje quirúrgico con lo que se obtuvo una buena evolución clínica. En el hemocultivo se aisló Haemophilus influenzae tipo b. Se destaca la importancia de la vacunación, así como de la vigilancia epidemiológica activa.

Summary: Haemophilus influenzae type b is a globally spread Gram-negative encapsulated bacterium. It is responsible for several invasive disease clinical manifestations, such as meningitis, epiglottitis, bacteremia, septic arthritis, cellulitis, osteomyelitis and pericarditis. These are immuno-preventable diseases. Facial cellulitis caused by Haemophilus influenzae type b is a potentially serious invasive infection predominantly affecting children under 2 years of age, since this an immunologically susceptible population. There are characteristic clinical elements, though not pathognomonic, that arise suspicion of this infection. We present a clinical case of an 8-month old infant, immunized with only one dose of BCG vaccine (calmetization), who arrived at the emergency ward with extended swelling of the face and was admitted to the intermediate care unit with a clinical diagnosis of cellulitis. Due to the rapid extension of the infectious process, he had to be transferred to the intensive care unit. Clinical tests to assess the infection and a CT scan were performed, which confirmed the process and the existence of a collection. We started empiric antibiotic therapy and surgical drainage and obtained a good clinical evolution. Haemophilus influenzae type b was isolated in the blood culture. We emphasize the importance of vaccination as well as of an active epidemiological surveillance.

Resumo: O Haemophilus influenzae tipo b é uma bactéria encapsulada Gram-negativa de disseminação global. É responsável por várias manifestações clínicas da doença invasiva, como meningite, epiglotite, bacteremia, artrite séptica, celulite, osteomielite e pericardite. Estas são doenças imunopreveníveis. A celulite do rosto causada por Haemophilus influenzae tipo b é uma infecção invasiva potencialmente grave que afeta predominantemente crianças menores de 2 anos de idade, uma vez que é uma população imunológicamente suscetível. Existem elementos clínicos característicos, embora não patognomônicos, que levantam suspeita dessa infecção. Apresentamos um caso clínico de um lactente de 8 meses de idade, imunizado com apenas uma dose da vacina BCG (calmetization), que chegou à sala de emergências com edema prolongado do rosto e foi admitido na unidade de cuidados intermediários com diagnóstico clínico de celulite. Devido à rápida extensão do processo infeccioso, ele teve que ser transferido para a unidade de terapia intensiva. Realizaram-se testes clínicos para avaliar a infecção e uma tomografia computadorizada, o que confirmou o processo e a existência de uma coleção. Iniciamos antibioticoterapia empírica e drenagem cirúrgica e obtivemos boa evolução clínica. O Haemophilus influenzae tipo b foi isolado na hemocultura. Ressaltamos a importância da vacinação e duma vigilância epidemiológica ativa.

Clinical characteristics of 84 children with invasive Haemophilus influenzae infection from 2014 to 2018 / 中华儿科杂志

Objective@#To investigate the clinical characteristics of invasive Haemophilus influenzae (HI) infection in children.@*Methods@#The clinical manifestations, laboratory examinations and treatment outcomes of 84 children with HI infection confirmed by bacterial culture in 7 tertiary children′s hospitals from 2014 to 2018 were analyzed retrospectively.@*Results@#Among the 84 cases, 50 were males. The age was 1.54 years (ranged from 5 days to 13 years).Twenty cases (24%) had underlying diseases and 48 cases (57%) had not received antibiotics before collecting specimens. Eighty-two cases (98%) had fever and 75 cases (89%) had clear infection foci, among which 31 cases (37%) had meningitis and 27 cases (32%) had pneumonia. Blood culture was positive in 62 cases (74%), cerebrospinal fluid culture was positive in 10 cases (12%), blood culture and cerebrospinal fluid culture were both positive in 11 cases (13%). Antibiotics susceptibility test showed that 27% (22/82) of all HI strains produced β-lactamases and 48% (37/77) strains were resistant to ampicillin. The drug resistance rates to cefuroxime, ampicillin-sulbactam, trimethoprim-sulfamethoxazole and azithromycin were 25% (20/80) , 20% (9/45) , 71% (44/62) and 19%(11/58), respectively. All strains were sensitive to meropenem, levofloxacin and ceftriaxone. After sensitive antibiotic therapy, 83% (70/84) of all patients were cured and improved, the mortality rate and loss of follow-up rate were 13% (11/84) and 4% (3/84) respectively.@*Conclusions@#Meningitis and pneumonia are common presentation of invasive HI infections in children. Mortality in HI meningitis children is high and the third generation of cephalosporins, such as ceftriaxone can be used as the first choice for the treatment of invasive HI infection.