Serotypes and antimicrobial resistance of Streptococcus pneumoniae in children under 5 years of age pre- and post-pneumococcal conjugate vaccine introduction in Paraguay.

Introduction. Streptococcus pneumoniae remains a major cause of mortality and morbidity worldwide in children <5 years of age, even with advances in vaccination programmes.Hypothesis/Gap Statement. Reviewing and reporting trends in the distribution of pneumococcal serotypes and antimicrobial resistance in Paraguay will be useful for decision-making in public health.Aim. This study analysed the serotype distribution and antimicrobial resistance of S. pneumoniae and the characteristics of pneumococcal disease in children <5 years old before and after the introduction of pneumococcal conjugate vaccines (PCVs).Methodology. A total of 885 isolates and 278 S. pneumoniae PCR-positive clinical specimens were referred to the Central Laboratory of Public Health (LCSP) within the meningitis and pneumonia laboratory based-surveillance network in the period 2006-2020. Conventional and molecular microbiological techniques were used for confirmation and characterization.Results. We identified 563 cases of pneumococcal disease in the pre-vaccination period, 325 cases in the post-PCV10 period and 275 cases in the post-PCV13 period. The serotypes covered by PCV10 decreased from 78.6-6.5 %. However, additional serotypes covered by PCV13 increased from 6.6-57.5% and non-PCV13 serotypes increased from 14.8-36.0 % (P<0.001) in the post-PCV13 period. In cases of meningitis, the rate of resistance to penicillin decreased after the introduction of conjugate vaccines. No resistance to ceftriaxone was found in any period. In cases without meningitis, the rate of resistance to penicillin and ceftriaxone decreased slightly. However, the rate of resistance to erythromycin and tetracycline increased and that to trimethoprim-sulfamethoxazole (SXT) decreased in the post-PCV13 period compared to the pre-PCV period. The multidrug resistance rate was 8.5 %.Conclusion. A change in the circulating serotypes and antimicrobial resistance to certain antibiotics was observed. Non-vaccine serotype circulation and multidrug resistance may compromise the success of the conjugate vaccines.

Serogrupos y resistencia antimicrobiana de Neisseria meningitidis causantes de enfermedad invasiva en Paraguay

RESUMEN La enfermedad meningocócica representa un problema de salud pública y una de las principales causas de morbilidad y mortalidad en todo el mundo. Los serogrupos que causan la mayor carga de enfermedad a nivel global son A, B, C, W e Y. El objetivo del estudio fue describir los serogrupos y la resistencia antimicrobiana de Neisseria meningitidis aisladas de enfermedad invasiva en Paraguay durante el periodo 2010-2020. Se estudiaron todas las muestras de líquido cefalorraquídeo y sangre con aislamientos o detección de ADN por PCR de N. meningitidis de pacientes de diversas edades, que fueron remitidas al Laboratorio Central de Salud Pública durante el periodo 2010-2020, dentro del marco de la vigilancia epidemiológica de meningitis (n=163) en Paraguay. La mayor frecuencia de hallazgos de N. meningitidis se observó en el grupo de edad de < 1 año. El 25,7% de los casos correspondió al serogrupo B, el 52,1% al serogrupo C, 18,4% al serogrupo W y 3,7% al serogrupo Y. En el 2018, se evidenció la mayor cantidad de casos por serogrupo C (n=27). La menor frecuencia de sensibilidad disminuida a penicilina G fue en el 2010 (12,5%) y la mayor en el 2014 (100,0%). Se registró un aumento de casos por serogrupo C a partir del 2017, posicionándose como serogrupo prevalente hasta el 2020, y además, un aumento de la sensibilidad disminuida a la penicilina. La vigilancia es de importancia en el control de la enfermedad meningocócica para detección de brotes, estimación de la carga de enfermedad, análisis de resistencia antimicrobiana, distribución de serogrupos y evaluaciones de estrategias de control.

ABSTRACT Meningococcal disease represents a public health problem and one of the main causes of morbidity and mortality worldwide. The serogroups that cause the highest burden of disease globally are A, B, C, W, and Y. The aim of the study was to describe serogroups and antimicrobial resistance of Neisseria meningitidis isolated from invasive disease in Paraguay during the 2010-2020 period. Samples of cerebrospinal fluid and blood with isolates or detection of DNA by PCR of N. meningitidis from patients of different ages referred to the Central Public Health Laboratory during the period 2010-2020 within the framework of the epidemiological surveillance of meningitis (n = 163) in Paraguay were studied. The highest frequency of N. meningitidis findings was observed in the <1 year age group, 25.7% of the cases corresponded to serogroup B, 52.1% to serogroup C, 18.4% to serogroup W and 3.7% to serogroup Y. In 2018, the highest number of cases by serogroup C (n = 27) was found. The lowest frequency of decreased sensitivity to penicillin G was in 2010 (12.5%) and the highest in 2014 (100.0%). There was an increase in cases due to serogroup C in 2017, positioning itself as the prevalent serogroup until 2020, in addition, there was an increase in decreased sensitivity to penicillin. Surveillance is important in the control of meningococcal disease for outbreak detection, estimation of the burden of disease, analysis of antimicrobial resistance, serogroup distribution, and evaluations of control strategies.

Estudio epidemiológico de Haemophilus influenzae causante de enfermedad invasiva y no invasiva en Paraguay (1999-2017) / Epidemiologic study of Haemophilus influenzae causing invasive and non-invasive disease in Paraguay (1999-2017)

Introducción: Haemophilus influenzae (Hi) causa infecciones leves e invasivas graves, especialmente en niños menores de 5 años. El serotipo b (Hib) era muy frecuente antes de la introducción de la vacuna, que en Paraguay se introdujo en 2004. Métodos: Se estudiaron por métodos microbiológicos convencionales y técnicas moleculares 523 aislados de H. influenzae obtenidos desde 1999 a 2017 y remitidos al Laboratorio Nacional de Referencia de Paraguay. Resultados: El serotipo más frecuente fue el no-tipable (HiNT), en un 51,8% (271/523), seguido de Hib (43%; 225/523), Hia e Hif (1,5%; 8/523, respectivamente), Hic (1%; 5/523), Hie (0,8%; 4/523) y Hid (0,4%; 2/523). El 48,4% de las infecciones invasivas estaban causadas por HiNT y el 46,1% por Hib. El 88,6% de las cepas procedían de casos de meningitis, el 70,8% de sepsis y el 50,9% de neumonías en niños menores de 5 años. El 84% (181/217) de las cepas correspondían a enfermedades invasivas causadas por Hib en menores de 5 años, registrándose la mayor proporción entre 2001-2003. Los biotipos más prevalentes fueron el biotipo I (29%), biotipo II (12%), biotipo III (24%) y biotipo IV (13%). El 13% (68/523) de los aislados fueron resistentes a la ampicilina. Conclusión: Tras de la introducción de la vacuna frente a Hib en Paraguay, los casos de enfermedad invasiva por Hib disminuyeron en niños menores de 5 años y hubo un aumento de HiNT en los mayores de 5 años. Es necesaria una vigilancia continua para monitorizar la eficacia de la vacuna y desarrollar intervenciones preventivas.(AU)

Introduction: Haemophilus influenzae is a cause of mild and severe invasive infections, especially among children under 5 years old. Serotype b (Hib) was very frequent before the introduction of the vaccine, which was introduced in Paraguay in 2004. Methods: A total of 523 isolates of H. influenzae obtained from 1999 to 2017 and referred to the National Reference Laboratory in Paraguay were studied by conventional microbiological methods and molecular techniques. Results: The most frequent serotype was non-typeable (HiNT) (51.8%; 271/523), followed by Hib (43%; 225/523), Hia and Hif (1.5%; 8/523, respectively), Hic (1%; 5/523), Hie (0.8%; 4/523), and Hid (0.4%; 2/523). A total of 48.4% invasive infections were caused by HiNT, and 46.1% by Hib; 88.6% of isolates corresponded to meningitis, 70.8% to sepsis and 50.9% to pneumonia in children under 5 years. A total of 84% (181/217) of isolates corresponded to invasive infections caused by Hib in children under 5 years, with the highest proportion observed between 2001 and 2003. The most prevalent biotypes were biotype I (29%), biotype II (12%), biotype III (24%), and biotype IV (13%). Among the total of isolates, 13% (68/523) of isolates were resistant to ampicillin. Conclusion: After the introduction of the Hib vaccine in Paraguay, the number of invasive Hib cases decreased in children under 5 years old, although we observed an increase of HiNT in children over 5 years. Continuous surveillance is necessary in order to monitor the effectiveness of the vaccine and for the development of preventive interventions.(AU)

Epidemiologic study of Haemophilus influenzae causing invasive and non-invasive disease in Paraguay (1999-2017). / Estudio epidemiológico de Haemophilus influenzae causante de enfermedad invasiva y no invasiva en Paraguay (1999-2017).

INTRODUCTION: Haemophilus influenzae is a cause of mild and severe invasive infections, especially among children under 5 years old. Serotype b (Hib) was very frequent before the introduction of the vaccine, which was introduced in Paraguay in 2004. METHODS: A total of 523 isolates of H. influenzae obtained from 1999 to 2017 and referred to the National Reference Laboratory in Paraguay were studied by conventional microbiological methods and molecular techniques. RESULTS: The most frequent serotype was non-typeable (HiNT) (51.8%; 271/523), followed by Hib (43%; 225/523), Hia and Hif (1.5%; 8/523, respectively), Hic (1%; 5/523), Hie (0.8%; 4/523), and Hid (0.4%; 2/523). A total of 48.4% invasive infections were caused by HiNT, and 46.1% by Hib; 88.6% of isolates corresponded to meningitis, 70.8% to sepsis and 50.9% to pneumonia in children under 5 years. A total of 84% (181/217) of isolates corresponded to invasive infections caused by Hib in children under 5 years, with the highest proportion observed between 2001 and 2003. The most prevalent biotypes were biotype I (29%), biotype II (12%), biotype III (24%), and biotype IV (13%). Among the total of isolates, 13% (68/523) of isolates were resistant to ampicillin. CONCLUSION: After the introduction of the Hib vaccine in Paraguay, the number of invasive Hib cases decreased in children under 5 years old, although we observed an increase of HiNT in children over 5 years. Continuous surveillance is necessary in order to monitor the effectiveness of the vaccine and for the development of preventive interventions.

Contaminación bacteriana de esponja de lavado y bachas de cocina de Cantinas de Escuelas de Asunción y Gran Asunción

RESUMEN Introducción: Las esponjas y el fregadero constituyen un medio favorable para el crecimiento de microorganismos. Objetivo: Evaluar la presencia de microorganismos en esponjas de lavado y bachas de cocina de cantinas de escuelas y colegios de Asunción y gran Asunción. Materiales y Métodos: estudio fue observacional descriptivo de corte transversal. Se tomó muestras de esponjas de lavado e hisopado de bachas de cocina de 25 cantinas de escuelas y colegios públicos y privados de Asunción y gran Asunción. Se utilizo medios de cultivo para bacterias aerobias mesófilas y coliformes fecales, Staphylococcus aureus y Salmonella spp. Se considero contaminación la presencia de ≥100UFC para las bacterias aerobias mesófilas y cualquier crecimiento de bacterias patógenas. El análisis se realizó en Micorsoft Excell utilizando estadísticas descriptivas. Resultados: Fueron incluidas 11 instituciones privadas y 14 públicas. El 88% (22/25 de las esponjas presentaron alta contaminación con bacterias aerobias mesófilas; 20 (80%) con coliformes fecales y 1 (4%) estéril. Hisopados de bachas, 21 (84%) presentaron alta contaminación las mesófilas y 20 (80%) con coliformes fecales, 1 (4%). No se aisló Salmonella spp. ni Staphylococcus aureus. No hubo diferencias en la contaminación de bachas entre instituciones públicas o privadas, pero si en las esponjas 14/14 vs 6/11 en las públicas y privadas respectivamente p=0.008 (T Fisher). Conclusión: esponjas de cocina y las bachas de las escuelas de Asunción y gran Asunción estudiadas, presentaron alto porcentaje de contaminación bacteriana tanto con bacterias aerobias mesófilas como con coliformes fecales. No se aisló Staphylococcus aureus ni Salmonella spp.

ABSTRACT Introduction: Sponges and the sink are a favorable medium for microorganism growth. Objective: To evaluate the presence of microorganisms in washing sponges and kitchen sinks in school cafeterias in Asunción and Greater Asunción. Materials and Methods: This was a descriptive, observational and cross-sectional study. Samples were taken by swabbing from washing sponges and kitchen sinks from 25 public and private school cafeterias in Asunción and the Greater Asunción area. Culture media were used for aerobic mesophilic and fecal coliform bacteria, Staphylococcus aureus and Salmonella species. The presence of ≥100 CFU for mesophilic aerobic bacteria and any growth of pathogenic bacteria was considered contamination. The analysis was performed with Microsoft Excel using descriptive statistics. Results: 11 private and 14 public institutions were included. 88% (22/25 of the sponges presented high contamination with aerobic mesophilic bacteria; 20 (80%) with fecal coliforms and 1 (4%) was sterile. Sink swabs: 21 (84%) presented high contamination with mesophylls and 20 (80%) with fecal coliforms, 1 (4%) was sterile. No Salmonella species or Staphylococcus aureus was isolated. There were no differences in the contamination of sinks between public or private institutions, but in sponges, 14/14 (public) were contaminated vs 6/11 (private), p = 0.008 (T Fisher). Conclusions: kitchen sponges and sinks from the schools of Asuncion and Greater Asuncion area in this study presented a high percentage of bacterial contamination with both aerobic mesophilic bacteria and fecal coliforms. No Staphylococcus aureus or Salmonella species were isolated.

Frecuencia de Streptococcus pneumoniae aislados de enfermedad invasiva en Paraguay, serotipos y perfil de sensibilidad (2010-2018) / Frequency of Streptococcus pneumoniae isolated from invasive disease in Paraguay, serotypes and sensitivity profile (2010-2018)

Streptococcus pneumoniae sigue siendo una de las causas más importantes de morbilidad y mortalidad en niños y adultos alrededor del mundo. El objetivo del estudio fue describir la frecuencia de aislamiento de S. pneumoniae en enfermedad invasiva, distribución de serotipos y sensibilidad antimicrobiana en Paraguay (2010-2018). Se estudiaron 793 cepas de S. pneumoniae aisladas de pacientes de todas las edades con enfermedad invasiva en Paraguay, provenientes de los diferentes centros centinelas y colaboradores en el marco de la vigilancia de meningitis y neumonías, durante el periodo 2010-2018. La frecuencia general según diagnóstico resultó 74.9% de neumonías (n=594), 18.4% de meningitis (n=146) y 6.7% de sepsis (n=53). El serotipo 14 fue más frecuente con 174 aislamientos (22.0%), seguido del serotipo 19A con 84 aislamientos (10.6%), el serotipo 3 con 66 aislamientos (8.3%) y el 6A con 37 aislamientos (4.7%). En meningitis se registró una frecuencia general de resistencia a penicilina del 32,2% y de ceftriaxona del 1,4%. En los casos de no meningitis la resistencia a penicilina fue del 0,8% y ceftriaxona del 0,3%. Los resultados de serotipos y sensibilidad antimicrobiana proporcionarán información necesaria para la implementación de estrategias de prevención y tratamiento de la enfermedad neumocócica en nuestro país, por lo que es necesaria una vigilancia continua para evaluar la carga de enfermedad, los serotipos circulantes y el aumento de la resistencia a los antibióticos

Streptococcus pneumoniae remains one of the most important causes of morbidity and mortality in children and adults worldwide. The objective of the study was to describe the frequency of isolation of S. pneumoniae in invasive disease, serotype distribution and antimicrobial susceptibility in Paraguay (2010-2018). We studied 793 strains of S. pneumoniae isolated from patients of all ages with invasive disease in Paraguay, from different sentinel centers and collaborators in the framework of meningitis and pneumonia surveillance during the period 2010-2018. The general frequency according to diagnosis was 74.9% of pneumonia (n = 594), 18.4% of meningitis (n = 146) and 6.7% of sepsis (n = 53). Serotype 14 was more frequent with 174 isolates (22.0%), followed by serotype 19A with 84 isolates (10.6%), serotype 3 with 66 isolates (8.3%) and 6A with 37 isolates (4.70%). In meningitis, there was a general frequency of penicillin resistance of 32.2% and ceftriaxone of 1.4%. In cases of non-meningitis, penicillin resistance was 0.8% and ceftriaxone 0.3%. The results of serotypes and antimicrobial sensitivity will provide necessary information for the implementation of prevention strategies and treatment of pneumococcal disease in our country, therefore it is necessary to continue monitoring in order to assess the burden of the disease, circulating serotypes and increased antibiotic resistance

Evaluación de la capacidad formadora de biofilm de cepas de S. aureus resistentes a meticilina que infectaron a niños paraguayos

Introducción: La capacidad de S. aureus para adherirse a superficies y a otras células bacterianas, para conformar un biofilm, es una de las principales causas de la persistencia de las infecciones por este patógeno. Objetivo: Analizar la capacidad de formación de biofilm, mediante el ensayo in vitro en placa de poliestireno, de cepas de S. aureus. Material y métodos: Estudio descriptivo de corte trasverso Se estudiaron 10 cepas resistentes a meticilina, con caracterización molecular de factores de virulencia y representativas de los grupos clonales detectados en un total de 347 cepas que causaron infecciones de piel y partes blandas e invasivas en niños en el 2012. La estandarización del ensayo in vitro fue exitosa, permitiendo clasificar a las cepas en 3 categorías según la intensidad de formación del biofilm. Resultados: El 100% de las cepas analizadas fue formadora de biofilm y se identificó dentro de la categoría de formadores moderados. Conclusiones: La detección de la formación de biofilms bacterianos, sobre todo en infecciones crónicas, es un tema crucial, ya que de ello depende la elección de estrategias especiales para su eliminación, como la remoción quirúrgica, dada la inefectividad de las terapias antibióticas tradicionales.

Introduction: The capacity of S. aureus to adhere to surfaces and other bacterial cells by forming a biofilm is one of the main causes of the persistence of infections by this pathogen. Objective: To analyze the capacity of S. aureus strains to form biofilm, using in vitro polystyrene plate culture techniques. Materials and methods: This was a descriptive, cross-sectional study. We studied and performed molecular characterization of viruence factors in ten strains of S. aureus, which were representative of the clonal groups detected in a total of 347 strains that caused skin and soft tissue invasive infections in children in 2012. The standardization of the in vitro assay was successful, allowing to classify the strains into 3 categories according to the intensity of biofilm formation. Results: 100% of the analyzed strains were biofilm formers and were categorized as moderate formers. Conclusions: The detection of the formation of bacterial biofilms, especially in chronic infections, is crucial, since it affects the choice of special strategies, such as surgical removal, for their elimination, given the ineffectiveness of traditional antibiotic therapies.

Síndrome diarreico agudo causado por Campylobacter spp. en pacientes menores de 11 años y su resistencia antimicrobiana a las drogas de elección para tratamiento 2010-2012, Paraguay / Acute Diarrhea Syndrome from Campylobacter spp. in Patients under Age 11 Years and Resistance to Drugs of Choice in Paraguay, 2010-2012

Objetivos: Determinar la prevalencia de Campylobacter spp. en pacientes menores de 11 años con síndrome diarreico agudo e indagar la resistencia antimicrobiana con respecto a las drogas de elección para el tratamiento clínico con Ciprofloxacina, Eritromicina y Tetraciclina. Materiales y Métodos: Se realizó un estudio descriptivo, retrospectivo de corte transverso, muestreo no probabilístico de casos consecutivos, cuya muestra tiene un tamaño de 1110 con un nivel de confianza de 95%. La población estudiada fue de pacientes pediátricos menores de 11 años cuyas muestras de heces fueron remitidas por las instituciones integrantes de la Red de Enteropatógenos al Laboratorio Central de Salud Pública en el periodo 2010-2012. Resultados: De 1110 muestras de heces estudiadas se aislaron Campylobacter spp. en 176 de ellas y corresponde al 16% de prevalencia. Se observó resistencia a las quinolonas en un 49 % de las muestras estudiadas, así como una resistencia a Tetraciclina del 28% y 1% de resistencia a los macrólidos (Eritromicina). Conclusión: Se observó una prevalencia importante de Campylobacter spp. como agente etiológico de síndrome diarreico agudo y es relevante la incorporación del aislamiento de este patógeno como agente etiológico rutinario en los análisis de coprocultivos realizados. La resistencia observada a las drogas de elección utilizadas como tratamiento nos obliga a tener en cuenta el control del uso indiscriminado de antimicrobianos

Spoligotyping of clinical Mycobacterium tuberculosis isolates from the state of Minas Gerais, Brazil.

We performed spoligotyping on 114 strains of the Mycobacterium tuberculosis (Mtb) complex that had been isolated from patients in Minas Gerais Health Units during 2004. A total of 82/114 (72%) clinical isolates were clustered and 32/114 (28%) were unique. Seven shared types containing nine strains were newly created. A total of nine patterns corresponded to unreported orphan strains, as evaluated against all of the strains recorded in the SITVIT2 proprietary database in the Institut Pasteur de la Guadeloupe. The major clades were composed of isolates that belong to the following genotypes: Latin-America and Mediterranean (63/114, 55.3%) (the ill-defined T superfamily) (12/114, 10.5%), Haarlem (8/114, 7%), X clade (6/114, 5.3%), S clade (3/114, 2.6%) and the East-African Indian and Manu types, each with 1/114 (0.9%) isolates. A considerable number of strains (n = 20, 17.5%) showed patterns that did not fall within any of the previously described major clades. We conclude the bulk of tuberculosis (TB) (92/114, 80.7%) in our location is recent evolutionary strains that belong to the principal genetic groups 2/3. Further studies on epidemiology of TB are required to understand Mtb biodiversity and TB transmission in this region.

Spoligotyping of clinical Mycobacterium tuberculosis isolates from the state of Minas Gerais, Brazil

We performed spoligotyping on 114 strains of the Mycobacterium tuberculosis (Mtb) complex that had been isolated from patients in Minas Gerais Health Units during 2004. A total of 82/114 (72 percent) clinical isolates were clustered and 32/114 (28 percent) were unique. Seven shared types containing nine strains were newly created. A total of nine patterns corresponded to unreported orphan strains, as evaluated against all of the strains recorded in the SITVIT2 proprietary database in the Institut Pasteur de la Guadeloupe. The major clades were composed of isolates that belong to the following genotypes: Latin-America and Mediterranean (63/114, 55.3 percent) (the ill-defined T superfamily) (12/114, 10.5 percent), Haarlem (8/114, 7 percent), X clade (6/114, 5.3 percent), S clade (3/114, 2.6 percent) and the East-African Indian and Manu types, each with 1/114 (0.9 percent) isolates. A considerable number of strains (n = 20, 17.5 percent) showed patterns that did not fall within any of the previously described major clades. We conclude the bulk of tuberculosis (TB) (92/114, 80.7 percent) in our location is recent evolutionary strains that belong to the principal genetic groups 2/3. Further studies on epidemiology of TB are required to understand Mtb biodiversity and TB transmission in this region.