Bordetella trematum infection: case report and review of previous cases.

BACKGROUND: Bordetella trematum is an infrequent Gram-negative coccobacillus, with a reservoir, pathogenesis, a life cycle and a virulence level which has been poorly elucidated and understood. Related information is scarce due to the low frequency of isolates, so it is important to add data to the literature about this microorganism. CASE PRESENTATION: We report a case of a 74-year-old female, who was referred to the hospital, presenting with ulcer and necrosis in both legs. Therapy with piperacillin-tazobactam was started and peripheral artery revascularization was performed. During the surgery, a tissue fragment was collected, where Bordetella trematum, Stenotrophomonas maltophilia, and Enterococcus faecalis were isolated. After surgery, the intubated patient was transferred to the intensive care unit (ICU), using vasoactive drugs through a central venous catheter. Piperacillin-tazobactam was replaced by meropenem, with vancomycin prescribed for 14 days. Four days later, levofloxacin was added for 24 days, aiming at the isolation of S. maltophilia from the ulcer tissue. The necrotic ulcers evolved without further complications, and the patient's clinical condition improved, leading to temporary withdrawal of vasoactive drugs and extubation. Ultimately, however, the patient's general condition worsened, and she died 58 days after hospital admission. CONCLUSIONS: Despite being a rare finding, B. trematum is typically associated with the clinical manifestation of disorders that predispose to ulcer development, which can be infected by microorganisms. The combination of antibiotic therapy and surgical debridement plays a key role in preventing systemic infections. Monitoring the appearance of new cases of B. trematum is essential, since it can be an emerging microorganism. Isolating and defining the clinical relevance of unusual bacteria yields a more accurate perspective in the development of new diagnostic tools and allows for assessment of proper antimicrobial therapy.

Prevalência e caracterização de cepas de bordetella pertussis deficientes em pertactina em país com vacina de células inteiras contra coqueluche / Prevalence and characterization of pertactin-deficient bordetella pertussis strains in a pertussis whole-cell vaccine country

Bordetella pertussis é o agente causal da coqueluche, importante problema de saúde pública em todo o mundo, com elevada morbidade e mortalidade em crianças menores de um ano de idade. Dois tipos de vacinas contra coqueluche estão disponíveis: vacinas de célula inteira (wP) baseada em suspensões de B. pertussis mortos e vacinas acelulares ( aP) baseada em um ou mais antígenos de B. pertussis altamente purificados, como a toxina, pertactina e fímbrias. Apesar de essas vacinas estarem disponíveis há mais de 50 anos, ainda hoje se observam ciclos epidêmicos da doença sugerindo que, embora a doença tenha diminuído, a transmissão de B. pertussis não está sendo controlada. Sabe-se que a duração da imunidade contra a coqueluche não é permanente e em comparação com a infecção natural, a duração da proteção após a vacinação parece ser menor. A proteção oferecida pelas vacinas aP parece ser menos duradoura que as wP, levantando a hipótese de que a substituição dessas vacinas pelas ap em muitos países, desde a década de 90, possa ter influenciado a reemergência da coqueluche. Diversos países relataram divergência antigênica das cepas circulantes em relação às cepas vacinais e enquanto as evoluções genéticas mais observáveis na era da vacina wP foram as variações alélicas e antigênicas, a aparição de cepas de B. pertussis deficientes em pertactina foi um fenômeno associado a introdução das vacinas acelulares. O objetivo do estudo foi verificar a ocorrência de deficiência de pertactina em 555 isolados de B. pertussis coletados no Brasil durante 2010-2016. Todos os 555 isolados foram caracterizados por PFGE e sorotipagem para determinar a distribuição dos perfis circulantes. O ensaio de ELISA foi realizado para triar os isolados de B. pertussis que não produzem pertactina. Todos os isolados identificados como deficientes em pertactina foram confirmados por PCR e immunoblotting e possíveis alvos genéticos para a deficiência foram determinadas pelo sequenciamento por Sanger. O sorotipo predominante, Fim3, vem sendo gradativamente substituído por Fim2 e Fim2,3, perfazendo 75% dos isolados em 2016. A tipagem por PFGE apresentou 110 perfis distintos, com seis perfis representando a maioria dos isolados testados. A triagem por ELISA identificou oito isolados deficientes em pertactina, mas apenas três foram confirmados por immunoblotting. A PCR indicou que um isolado tinha uma mutação na região promotora do gene prn, enquanto os outros dois não apresentaram uma explicação genética óbvia para a sua deficiência. O sequenciamento revelou que os oito isolados carregavam o alelo prn2, o tipo mais prevalente em isolados recentes. Embora a deficiência de pertactina tenha sido identificada em alguns isolados, este estudo não detectou uma ocorrência relevante de deficiência de pertactina, confirmando observações prévias de que essa deficiência é provavelmente impulsionada pelas vacinas acelulares. (AU)

Bordetella pertussis is the causative agent of pertussis, an important public health concern worldwide, with high morbidity and mortality in infants. Two types of pertussis vaccines are available: whole-cell (wP) vaccines based on killed B. pertussis organisms, and acellular (aP) vaccines based on one or more highly purified individual pertussis antigens like pertussis toxin, pertactin and fimbriae. Despite the availability of these safe and effective vaccines for more 50 years old, there are still epidemic cycles of the disease suggesting that although disease was decreased, transmission of B. pertussis is not being controlled. It is known that the duration of immunity against pertussis is not permanent and compared to a natural infection, the duration of protection after vaccination appears to be shorter. The protection provided by aP is less enduring raising the possibility that the switch from wP vaccines to aP vaccines in many countries since the 1990s may have aggravated the pertussis ressurgence. Several countries reported antigenic divergence between circulating strains and vaccine strains and while the most observable genetic evolutions in the wP vaccine were allelic and antigenic variations, the emergence of pertactin deficient strains was a phenomenon associated with the introduction of acellular vaccines. The aim of this study was to determine the occurrence of pertactin deficiency in 555 isolates of B. pertussis collected in Brazil during 2010-2016. All isolates were characterized by PFGE and serotyping to determine the distribution of circulating profiles. ELISA screening was done to detect pertussis isolates not producing pertactin. All pertactin deficient isolates identified were confirmed by immunoblotting and PCR, and possible genetic sources for the deficiency were determined by Sanger sequencing. The predominant serotype, Fim3, has been gradually replaced by Fim2 and Fim2,3, making up 75% of the isolates in 2016. PFGE typing showed 110 profiles, with six profiles representing most of the isolates tested. ELISA screening identified eight pertactin deficient isolates, but only three were confirmed by immunoblotting. PCR indicated that one isolate had a promoter mutation in prn, while the other two did not have an obvious genetic explanation for their deficiency. Sequencing revealed that the eight strains carried the prn2 allele, the most prevalent type in modern isolates. While pertactin deficiency was identified in a few isolates, this study did not detect a relevant occurrence of pertactin deficiency, confirming previous observations that pertactin deficiency is likely aP driven. (AU)

The classical Bordetella species and MALDI-TOF technology: a brief experience.

PURPOSE: The aim of this work was to evaluate and optimize the identification of Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica (usually known as the classical Bordetella species) using Bruker Biotyper matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF MS). METHODOLOGY: A set of 106 previously characterized clinical isolates was used. The results were interpreted according to the manufacturer's recommendations and, in addition, a new score value cutoff was used for species identification. Further, the 10 % rule (previously adopted by other authors) and the new 5 % breaking point (proposed in this work) were evaluated in order to optimize identification rates.Results/Key findings. Our results suggest that it is possible to distinguish different species of the classical Bordetella species by following a simple algorithm without additional testing being required. CONCLUSION: MALDI-TOF might be a reliable tool for the identification of this group of bacteria when a combination of cutoff scores is used. This procedure allows us to increase the identification rates for the classical Bordetella species significantly; however, more studies will be required to determine the applicability of the method to other difficult-to-distinguish organisms.

[Features of Bordetella pertussis, Bordetella spp. infection and whopping cough in Córdoba province, Argentina]. / Caracterización de la infección por Bordetella pertussis, Bordetella spp. y coqueluche en la provincia de Córdoba, Argentina.

INTRODUCTION: Whooping cough is a re-emerging infection in the world and Latin America. OBJECTIVE: It was considered relevant to investigate the clinical and epidemiological profile of Bordetella spp. and Bordetella pertussis infection in Córdoba province, Argentina; evaluating, at the same time, the co-infection with virus producing respiratory infections that may be confused with whooping cough. MATERIAL AND METHODS: All whooping cough suspected cases were studied by Polimerase Chain Reaction, amplifying the repeated insertion sequence (IS) 481 and the promoter gene encoding pertussis toxin, between 2011 and 2013. The data were obtained from the clinical and epidemiological records. RESULTS: From 2,588 whooping cough suspected cases, 11.59% was infected by Bordetella spp. and 9.16% was confirmed as Bordetella pertussis infection. The rate of infection was 7.22 and 1.84 per 100,000 for 2011 and 2012, respectively. The infection presented a seasonal tendency and it was mainly found on the group of children between 13 and 24 months old. The co-infection with virus producing respiratory infections, were uncommon. Paroxysmal cough, cyanosis and/or vomiting were predictors of the infection for Bordetella pertussis. DISCUSSION AND CONCLUSIONS: To deal with the re-emergence of whooping cough is important the knowledge of the regional epidemiological situation. This paper shows the situation of these infections in the regional clinical and epidemiological context, and makes the information available for health decision-making.

Caracterización de la infección por Bordetella pertussis, Bordetella spp. y coqueluche en la provincia de Córdoba, Argentina / Features of Bordetella pertussis, Bordetella spp. infection and whopping cough in Córdoba province, Argentina

Introduction: Whooping cough is a re-emerging infection in the world and Latin America. Objective: It was considered relevant to investigate the clinical and epidemiological profile of Bordetella spp. and Bordetella pertussis infection in Córdoba province, Argentina; evaluating, at the same time, the co-infection with virus producing respiratory infections that may be confused with whooping cough. Material and Methods: All whooping cough suspected cases were studied by Polimerase Chain Reaction, amplifying the repeated insertion sequence (IS) 481 and the promoter gene encoding pertussis toxin, between 2011 and 2013. The data were obtained from the clinical and epidemiological records. Results: From 2,588 whooping cough suspected cases, 11.59% was infected by Bordetella spp. and 9.16% was confirmed as Bordetella pertussis infection. The rate of infection was 7.22 and 1.84 per 100,000 for 2011 and 2012, respectively. The infection presented a seasonal tendency and it was mainly found on the group of children between 13 and 24 months old. The co-infection with virus producing respiratory infections, were uncommon. Paroxysmal cough, cyanosis and/or vomiting were predictors of the infection for Bordetella pertussis. Discussion and Conclusions: To deal with the re-emergence of whooping cough is important the knowledge of the regional epidemiological situation. This paper shows the situation of these infections in the regional clinical and epidemiological context, and makes the information available for health decision-making.

Introducción: Coqueluche es una enfermedad reemergente en el mundo y en Latinoamérica. Objetivo: Resultó de interés caracterizar el perfil clínico-epidemiológico de la infección por Bordetella spp. y Bordetella pertussis en Córdoba, Argentina; evaluando además, la frecuencia de infecciones de etiología viral que, por cursar con un síndrome coqueluchoide (SC), pueden ser confundidas con cuadros de coqueluche. Material y Métodos: Los casos sospechosos de coqueluche, se estudiaron por reacción de polimerasa en cadena; amplificando la secuencia repetida de inserción (IS) 481 y la región promotora del gen de la toxina pertussis; entre 2011 y 2013. Los datos de los pacientes se obtuvieron de las fichas clínicoepidemiológicas. Resultados: De 2.588 pacientes, 11,59% presentó una infección por Bordetella spp. y en 9,16% se confirmó una infección por Bordetella pertussis. La tasa de infección fue 7,22 y 1,84 por 100.000 habitantes en 2011 y 2012, respectivamente. La infección presentó una tendencia estacional y se concentró principalmente en niños entre 13 y 24 meses. La tos paroxística, cianosis y/o vómitos fueron predictores de la infección por B. pertussis. La coinfección con virus productores de infecciones respiratorias fue poco frecuente. Discusión y Conclusiones: Es fundamental el conocimiento de la situación epidemiológica regional. Este trabajo presenta la situación de Córdoba y pone a disposición de la comunidad sanitaria la información para la toma de decisiones en el contexto clínico-epidemiológico regional.

Isolamento e identificação de Pasteurella multocida e Bordetella spp. em coelhos (Oryctolagus cuniculus) / Isolation and identification of Pasteurella multocida and Bordetella spp. in rabbits (Oryctolagus cuniculus)

As doenças respiratórias representam uma das principais causas de morbidade e mortalidade em coelhos. A pasteurelose é a doença bacteriana de maior impacto na cunicultura brasileira. Infecções mistas por Pasteurella multocida e Bordetella spp. resultam em um quadro clínico grave, com descarte de matrizes e elevado prejuízo econômico. O objetivo desse trabalho foi pesquisar a presença de Pasteurella multocida e Bordetella spp. em uma granja de coelhos comerciais com histórico de doença respiratória crônica. Foram avaliados 34 animais doentes. Na cultura bacteriológica foram identificados 6 animais positivos para Pasteurella multocida. e 3 positivos para Bordetella spp. Estes achados confirmam a presença destes agentes na granja e apontam a necessidade de adoção de medidas sanitárias para controle da infecção.(AU)

Respiratory diseases represent a major cause of morbidity and mortality in rabbits. Pasteurellosis is the most important bacterial disease on Brazilian cuniculture flocks. The infection associated by P. multocida and Bordetella spp. results in a severe clinical picture, and becomes responsible for waste matrices and high economic damage. The aim of this study was to search the presence of Pasteurella multocida and Bordetella spp in a commercial farm of rabbits, with a history of chronic respiratory disease. Thirty-four sick animals were investigated. The results of bacteriological tests showed six animals positive for Pasteurella multocida and 3 positive for Bordetella spp. These results confirm the presence of both pathogens and highlight the need of sanitary management for infection control.(AU)

Isolamento e identificação de Pasteurella multocida e Bordetella spp. em coelhos (Oryctolagus cuniculus) / Isolation and identification of Pasteurella multocida and Bordetella spp. in rabbits (Oryctolagus cuniculus)

As doenças respiratórias representam uma das principais causas de morbidade e mortalidade em coelhos. A pasteurelose é a doença bacteriana de maior impacto na cunicultura brasileira. Infecções mistas por Pasteurella multocida e Bordetella spp. resultam em um quadro clínico grave, com descarte de matrizes e elevado prejuízo econômico. O objetivo desse trabalho foi pesquisar a presença de Pasteurella multocida e Bordetella spp. em uma granja de coelhos comerciais com histórico de doença respiratória crônica. Foram avaliados 34 animais doentes. Na cultura bacteriológica foram identificados 6 animais positivos para Pasteurella multocida. e 3 positivos para Bordetella spp. Estes achados confirmam a presença destes agentes na granja e apontam a necessidade de adoção de medidas sanitárias para controle da infecção.

Respiratory diseases represent a major cause of morbidity and mortality in rabbits. Pasteurellosis is the most important bacterial disease on Brazilian cuniculture flocks. The infection associated by P. multocida and Bordetella spp. results in a severe clinical picture, and becomes responsible for waste matrices and high economic damage. The aim of this study was to search the presence of Pasteurella multocida and Bordetella spp in a commercial farm of rabbits, with a history of chronic respiratory disease. Thirty-four sick animals were investigated. The results of bacteriological tests showed six animals positive for Pasteurella multocida and 3 positive for Bordetella spp. These results confirm the presence of both pathogens and highlight the need of sanitary management for infection control.

Glycomacropeptide administration attenuates airway inflammation and remodeling associated to allergic asthma in rat.

OBJECTIVE: Glycomacropeptide (GMP) is a bioactive peptide derived from milk that has been reported to exhibit a range of anti-inflammatory and immunomodulatory properties. The aim of this study was to analyze the prophylactic effect of GMP administration on airway inflammation and remodeling in an experimental model of asthmatic rat. METHODS: Animals treated orally with or without GMP (500 mg/kg/day) were ovalbumin-sensitized and -nebulized and several indicators of Th2 response, airway structural changes and inflammatory cells recruitment were evaluated. RESULTS: Treatment with GMP prior and during asthma development resulted in reduction of allergen-specific IgE titers in serum and blood eosinophilia. Also, GMP substantially suppressed the recruitment of inflammatory cells to bronchoalveolar compartment. Histological studies demonstrated that GMP markedly inhibits eosinophils infiltration, goblet cells hyperplasia and collagen deposit in lung tissue. The latter effect was related with an inhibition in transforming growth factor-ß expression. In addition, expression of interleukin-5 and -13 were substantially inhibited in lung while that of interleukin-10 was increased. CONCLUSION: Our results suggest that administration of GMP may prevent the development of an excessive Th2 response in asthma and effectively ameliorates the progression of the disease.

[Presence of Bordetella holmesii in an outbreak of pertussis in Chile]. / Presencia de Bordetella holmesii en brote epidémico de coqueluche en Chile.

The incidence of whooping cough in Chile ranges from 4.1 and 7.5 per hundred thousand inhabitants. B. pertussis detection is performed by Real Time PCR (Q-PCR) directed to the insertion sequence IS481. However, this sequence is also found in the genome of B. bronchiseptica and B. holmesii. The latter is also a respiratory pathogen whose clinical features are similar to B. pertussis. However, it is important to differentiate between these species because in immunosuppressed patients B. holmesii is more likely to cause bacteremia and is less susceptible to erythromycin. The goal of this work is to measure prospectively and retrospectively the presence of B. holmesii in samples reported positive for B. pertussis in the period 2010-2011. During this period, 1994 nasopharyngeal specimens entered the laboratory for Bordetella sp. PCR, of which 224 were positive. The analysis by Q-PCR directed to the recA gene of B. holmesii of all 224 positive samples determined a prevalence of B. holmesii of 0.6% (12/1994). Because of its more aggressive behavior in immunosupressed patients and its different resistance pattern, routine screening of B. pertussis and B. holmesii is currently performed for all samples in which Bordetella sp PCR is initially detected.

Presencia de Bordetella holmesii en brote epidémico de coqueluche en Chile / Presence of Bordetella holmesii in an outbreak of pertussis in Chile

The incidence of whooping cough in Chile ranges from 4.1 and 7.5 per hundred thousand inhabitants. B. pertussis detection is performed by Real Time PCR (Q-PCR) directed to the insertion sequence IS481. However, this sequence is also found in the genome of B. bronchiseptica and B. holmesii. The latter is also a respiratory pathogen whose clinical features are similar to B. pertussis. However, it is important to differentiate between these species because in immunosuppressed patients B. holmesii is more likely to cause bacteremia and is less susceptible to erythromycin. The goal of this work is to measure prospectively and retrospectively the presence of B. holmesii in samples reported positive for B. pertussis in the period 2010-2011. During this period, 1994 nasopharyngeal specimens entered the laboratory for Bordetella sp. PCR, of which 224 were positive. The analysis by Q-PCR directed to the recA gene of B. holmesii of all 224 positive samples determined a prevalence of B. holmesii of 0.6% (12/1994). Because of its more aggressive behavior in immunosupressed patients and its different resistance pattern, routine screening of B. pertussis and B. holmesii is currently performed for all samples in which Bordetella sp PCR is initially detected.

La incidencia de coqueluche en Chile varía entre 4,1 y 7,5 por 100.000 habitantes. La detección de Bordetella pertussis se realiza por RPC-tiempo real (Q-RPC) dirigida a la secuencia de inserción IS481. Sin embargo, esta secuencia se encuentra también en el genoma de B. bronchiseptica y B. holmesii. Este último es también un patógeno respiratorio que produce un cuadro similar a B. pertussis. Sin embargo, es importante diferenciar entre estas especies porque en pacientes inmunosuprimidos B. holmesii tiene mayor tendencia a causar bacteriemia y además es menos susceptible a eritromicina. El objetivo de este trabajo es determinar, prospectiva y retrospectivamente, la presencia de B. holmesii en muestras informadas positivas para B. pertussis en el período 2010-2011. Durante ese período ingresaron al laboratorio 1. 994 muestras de hisopado nasofaríngeo para RPC de Bordetella sp., de las cuales 224 fueron positivas. El análisis por Q-RPC dirigido al gen recA de B. holmesii de las 224 muestras positivas determinó una prevalencia de B. holmesii de 0,6% (12/1994). Debido al comportamiento más agresivo en inmunosuprimidos y al patrón de resistencia de B. holmesi, se decide incorporar la detección de rutina de B. pertussis y B. holmesii en todas las muestras en que se detecta inicialmente la presencia de Bordetella sp.

Bordetella holmesii in children suspected of pertussis in Argentina.

We describe nine patients (eight aged <1 year) clinically diagnosed with pertussis yet laboratory-confirmed with Bordetella holmesii infections, a human pathogen normally isolated from blood. Most patients reported cough and cold symptoms. No death was reported. We report B. holmesii isolation in infants with respiratory symptoms in Argentina.

Desarrollo y validación de una PCR múltiple para el diagnóstico de Bordetella spp / Development and validation of a multiplex PCR for diagnosis of Bordetella spp / Desenvolvimento e validação de uma PCR múltipla para o diagnóstico de Bordetella spp

El objetivo del trabajo consistió en diseñar y validar una PCR en formato convencional que permita confirmar la presencia o ausencia de Bordetella pertussis y detectar otras especies del género, como Bordetella parapertussis y Bordetella bronchiseptica, que pudieran estar involucradas en el cuadro clínico de coqueluche. A tal fin se diseñó una reacción en cadena de la polimerasa (PCR) múltiple que amplifica una secuencia del promotor del gen de Toxina Pertussis y otra del gen de la Toxina Adenilato Ciclasa-Hemolisina. Se validó la metodología siguiendo esquemas publicados anteriormente. Se optimizaron las condiciones de la PCR. Se validó la metodología obteniéndose un límite de detección para ambas secuencias de 0,5 bacterias por reacción. Se validó, además, la especificidad y robustez de la técnica. Se presenta una nueva herramienta diagnóstica optimizada y validada, que permite detectar la presencia de las especies de Bordetella más frecuentemente involucradas en el cuadro clínico de coqueluche. Su uso combinado con alguna de las PCR habituales en diagnóstico, como la PCR IS481, permite aumentar la sensibilidad del diagnóstico de esta en­fermedad, la especificidad del mismo discriminando los resultados falsos positivos/negativos y aumentar el conocimiento sobre los agentes etiológicos implicados en esta patología.

The aim of the present work was to design and validate a conventional PCR that enables to confirm the presence or absence of Bordetella pertussis and to detect other Bordetella species, such as Bordetella parapertussis metoand B. bronchiseptica, that may be involved in this pathology. To this aim, a multiplex PCR that amplifies a sequence of the promoter of the Pertussis Toxin gene and a sequence of the Adenylate Cyclase Toxin-Hemolysin gene were designed. The PCR was validated following previously published schemes. PCR conditions were optimized. The methodology was validated obtaining a detection limit of 0.5 bacteria per reaction, for both sequences. Specificity and robustness of the technique were also validated. A new optimized and validated tool to detect the presence of the Bordetella species most frequently responsible of pertussis was presented. The combined use with some of the usual PCR, such as IS 481, may increase the sensitivity of the diagnosis of this disease, its specificity discriminating false positive/negative results and increase awareness of the etiologic agents involved in this pathology.

O objetivo do trabalho foi desenhar e validar uma reação em cadeia da polimerase (PCR) em formato convencional que permita confirmar a presença ou ausência de Bordetella pertussis e detectar outras espécies do gênero, como Bordetella parapertussis e Bordetella bronchiseptica, que pudessem estar envolvidas no quadro clínico de coqueluche. Para tal, foi desenhada uma PCR múltipla que amplifica uma sequência do promotor do gene de Toxina Pertussis e outra do gene da Toxina Adenilato Ciclase-Hemolisina. A metodologia foi validada seguindo esquemas publicados anteriormente. Foram otimizadas as condições da PCR. Validou-se a metodologia obtendo-se um limite de detecção para ambas as sequências de 0,5 bactérias por reação. Validou-se também a especificidade e robustez da técnica. Apresenta-se uma nova ferramenta diagnóstica otimizada e validada, que permite detectar a presença das espécies de Bordetella mais frequentemente envolvidas no quadro clínico de coqueluche. Seu uso combinado com alguma das PCR habituais em diagnóstico, como a PCR IS481, permite aumentar a sensibilidade do diagnóstico desta doença, a especificidade do mesmo discriminando os resultados falsos positivos/negativos e aumentar o conhecimento sobre os agentes etiológicos envolvidos nesta patologia.

Caracterização fenotípica e gonotípica de Bordetella pertussis

Na era pré-vacinação, a coqueluche representou uma das principais causas da morbidade e mortalidade infantil. Significativa diminuição na sua incidência é verificada com a introdução da vacinação na década de 40. No entanto, a partir da década de 80, a coqueluche passou novamente a representar um importante problema de saúde pública em vários países, sendo comum a ocorrência de epidemias cíclicas a cada de 3 - 5 anos. Atualmente, a coqueluche é classificada pelo Centro de Controle de Doenças nos Estados Unidos (CDC) como uma doença re-emergente. As causas deste aumento não são conhecidas; no entanto podem estar associadas à imunidade induzida pela vacina. A coqueluche, causada pela Bordetella pertussi, é uma doença respiratória aguda, altamente contagiosa, e especialmente severa em crianças menores. Embora seja uma doença predominantemente da infância, nos últimos anos, tem sido reportado um aumento significativo nos adolescentes e adultos. O objetivo deste estudo foi caracterizar 72 cepas de B. pertussis isoladas da secreção nasofaríngea de casos suspeitos de coqueluche. Para o isolamento foi utilizado...

Detection of Bordetella bronchiseptica by the polymerase chain reaction.

Polymerase chain reaction (PCR) assays were developed that enabled not only discriminative detection of three Bordetella species, B. pertussis, B. parapertussis, and B. bronchiseptica (Bspp PCR), but also specific detection of B. bronchiseptica (Bb PCR). An upstream sequence of the flagellin gene was used as a target DNA region. This sequence contained differences in B. pertussis, B. parapertussis, and B. bronchiseptica DNA. These species could then be differentiated using two different sets of primers, Bspp and Bb. When oligonucleotide Bspp primers were used, PCR products were obtained from the three species of Bordetella. A fragment of the expected size (164 bp) was amplified using B. bronchiseptica and B. parapertussis DNA, but a fragment with a distinct molecular weight was amplified with B. pertussis DNA (195 bp). This Bspp PCR was specific and sensitive, but it could not differentiate between B. parapertussis and B. bronchiseptica. When Bb primers were used, a 237-bp PCR product was detected only from B. bronchiseptica DNA. No PCR products were identified after Bb PCR amplification of DNAs either from B. parapertussis isolates or B. pertussis isolates, nor from other respiratory pathogen DNAs tested. This second PCR assay had a sensitivity limit of less than 10 organisms of B. bronchiseptica after detection with a specific probe. The specificity and the sensitivity of the fla PCR assay were evaluated with purified DNA, as was its capacity for detecting the bacteria in human clinical samples and in lungs of infected mice.

Biological and protective activities of outer membrane proteins from Bordetella.

In this work the biological properties and the capability of the outer membrane proteins (OMPs) from different strains of Bordetella to induce protection against challenge with B. pertussis 18323 were examined. The OMPs from each strain were isolated using Schnaitmann's method. Two OMPs (30 and 32 kDa) were found to be specific for the vaccine strains of B. pertussis and were absent in the OMPs preparation from both B. parapertussis and B. bronchiseptica. When the OMPs from the vaccine strains of B. pertussis were assayed in the mouse intracerebral protection test, they were found to be highly protective (75%-88%) against a challenge with 250 50% lethal doses (LD50) of B. pertussis 18323. However, no correlation was observed between the protective activity and the lymphocytosis-promoting factor (LPF) content of different preparations. Moreover, neither LPF activity or histamine-sensitizing activity (HSA) were found in any of the OMPs assayed. Our results show that OMPs from B. pertussis vaccine strains play a key role in the induction of protective immunity against B. pertussis 18323 in mice, making them excellent candidates to be used in further studies for the development of a pertussis vaccine for humans.

Prevalencia de Bordetella en niños de la ciudad de Lima / Prevalence of Bordetella in children in Lima

Se estudiaron 287 hisopados nasofaríngeos, procedentes de niños entre 1 mes y 12 años de edad, atendidos en el Hospital del Niño, ciudad de Lima. Dichos pacientes, tenían en su mayor parte diagnóstico de tos ferina. Las muestras tomadas se sembraron en el medio de Bordet-Gengou, identificándose las colonias por características culturales, pruebas bioquímicas y/o aglutinaciones. Se aislaron en 5 casos cepas de Bordetella, lo que representa el 17.86% de las muestras estudiadas. Dos de las cepas pudieron ser identificadas como B. parapertussis y B. pertussis, respectivamente. Se pone de manifiesto la importancia del aislamiento de B. parapertussis de un paciente con síntomas típicos de tos ferina.

28 pharyngeal swabs were studied in children who attended to the Children Hospital from Lima. Most of children had clinical diagnosis of whooping cough. The samples were seeded on Bordet Gengou Medium and identified by morphology culture, physiological characteristics, and agglutination test. Five strains of Bordetella were isolated. Two strains were identified as B. parapertussis and B. pertussis, respectively. It was indicated the importance of B. parapertussis isolation from a patient with typical whooping cough symptoms.

Increase of the bursal follicular medullary compartment in chicks intracloacally inoculated with Bordetella pertussis.

The cloacal bursa is a primary lymphoid organ responsible for the maturation of B-lymphocytes. It has been suggested that the bursa may also play a peripheral role when antigens are inoculated by cloacal route. Qualitative and quantitative structural modifications in the bursa from chicks inoculated with Bordetella pertussis by the cloacal route were investigated. Observations indicated that the relative bursal growth as well as the volume fraction and the mitotic index of the follicular medulla from experimental bursae are significantly greater than those of the controls. Macrophages which have phagocytized bacteria, and a gradual relative increase of the RER of lymphoblasts, were other structural modifications found exclusively in the follicular medulla. The observations suggest that the bursal follicular cortex and medulla act as autonomous histophysiological compartments, the latter being responsible for an antigenic stimulation when Bordetella pertussis is intracloacally inoculated in chicks.