Bordetella bronchiseptica is a potent and safe adjuvant that enhances the antigen-presenting capability of dendritic cells

We previously demonstrated that Bordetella bronchiseptica (B. bronchiseptica) antigen (Ag) enhances the Mycoplasma hyopneumoniae Ag-specific immune response. The focus of this study was whether acellular bacterin of B. bronchiseptica could be used as an adjuvant to increase antigen-presenting capability of dendritic cells (DCs) by increasing the level of activation. The metabolic activity of DCs was increased by B. bronchiseptica, similar to lipopolysaccharide (LPS). Flow cytometry analysis revealed that B. bronchiseptica increases the expression of major histocompatibility complex class-2, cluster of differentiation (CD)40, CD54, and CD86 which are closely related to DC-mediated immune responses. B. bronchiseptica enhanced the production of cytokines related to adaptive immune responses. Furthermore, the survival rate of B. bronchiseptica-injected groups was 100% at 15 and 20 mg/kg doses, whereas that of LPS-injected groups was only 20%, 0% at 15 and 20 mg/kg doses respectively, and so B. bronchiseptica is likely to be safer than LPS. Taken together, these results indicate that B. bronchiseptica can be used as an adjuvant to enhance the antigen-presenting capability of DCs. B. bronchiseptica is a candidate for producing vaccines, especially in case of DC-mediating efficacy and safety demands. This study provides researchers and clinicians with valuable information regarding the usage of B. bronchiseptica as a safe bacteria-derived immunostimulating agent for developing efficient vaccines.

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.

Bordetella bronchiseptica antigen enhances the production of Mycoplasma hyopneumoniae antigen-specific immunoglobulin G in mice

We previously demonstrated that Bordetella (B.) bronchiseptica antigen (Ag) showed high immunostimulatory effects on mouse bone marrow cells (BMs) while Mycoplasma (M.) hyopneumoniae Ag showed low effects. The focus of this study was to determine if B. bronchiseptica Ag can enhance the M. hyopneumoniae Ag-specific immune response and whether the host's immune system can recognize both Ags. MTT assay results revealed that each or both Ags did not significantly change BM metabolic activity. Flow cytometry analysis using carboxyfluorescein succinimidyl ester showed that B. bronchiseptica Ag can promote the division of BMs. In cytokine and nitric oxide (NO) assays, B. bronchiseptica Ag boosted production of tumor necrosis factor-alpha in M. hyopneumoniae Ag-treated BMs, and combined treatment with both Ags elevated the level of NO in BMs compared to that from treatment of M. hyopneumoniae Ag alone. Immunoglobulin (Ig)G enzyme-linked immunosorbent assay using the sera of Ag-injected mice clearly indicated that B. bronchiseptica Ag can increase the production of M. hyopneumoniae Ag-specific IgG. This study provided information valuable in the development of M. hyopneumoniae vaccines and showed that B. bronchiseptica Ag can be used both as a vaccine adjuvant and as a vaccine Ag.

Drug Target Identification and Elucidation of Natural Inhibitors for Bordetella petrii: An In Silico Study

Environmental microbes like Bordetella petrii has been established as a causative agent for various infectious diseases in human. Again, development of drug resistance in B. petrii challenged to combat against the infection. Identification of potential drug target and proposing a novel lead compound against the pathogen has a great aid and value. In this study, bioinformatics tools and technology have been applied to suggest a potential drug target by screening the proteome information of B. petrii DSM 12804 (accession No. PRJNA28135) from genome database of National Centre for Biotechnology information. In this regards, the inhibitory effect of nine natural compounds like ajoene (Allium sativum), allicin (A. sativum), cinnamaldehyde (Cinnamomum cassia), curcumin (Curcuma longa), gallotannin (active component of green tea and red wine), isoorientin (Anthopterus wardii), isovitexin (A. wardii), neral (Melissa officinalis), and vitexin (A. wardii) have been acknowledged with anti-bacterial properties and hence tested against identified drug target of B. petrii by implicating computational approach. The in silico studies revealed the hypothesis that lpxD could be a potential drug target and with recommendation of a strong inhibitory effect of selected natural compounds against infection caused due to B. petrii, would be further validated through in vitro experiments.

A Bordetella pertussis proteoliposome induces protection in mice without affecting the immunogenicity of diphtheria and tetanus toxoids in a trivalent formulation

In this study, a formulation of Bordetella pertussis proteoliposome (PLBp), diphtheria, and tetanus toxoids and alum (DT-PLBp) was evaluated as a trivalent vaccine candidate in BALB/c mice. Vaccine-induced protection was estimated using the intranasal challenge for pertussis and enzyme-linked immunosorbent assay fvto assess serological responses for diphtheria or tetanus. Both, diphtheria-tetanus-whole cell pertussis (DTP) and diphtheria-tetanus vaccines (DT) were used as controls. Animals immunized with DT-PLBp, PLBp alone, and DTP showed total reduction of CFU in lungs 7 days after intranasal challenge. Likewise, formulations DT-PLBp, DTP, and DT elicited antibody levels ≥2 IU/mL against tetanus and diphtheria, considered protective when neutralization tests are used. Overall, results showed that combination of PLBp with tetanus and diphtheria toxoids did not affect the immunogenicity of each antigen alone.

Bordetella bronchiseptica Respiratory Infection in the Immunosuppressed Patient / 대한내과학회지

Bordetella bronchiseptica is a common cause of respiratory disease in animals but is a rare cause of human infection. Furthermore, most patients with Bordetella bronchiseptica infections are immunocompromised. The Bordetella bronchiseptica organism can cause pneumonia, septicemia, and peritonitis in humans with impaired immune systems. Additionally, it can lead to a life-threatening infection patients who have an underlying debilitation or impaired immunity. The respiratory tract is the most common site of infection. Sixty-two human cases of Bordetella bronchiseptica have been published in the English literature, and 84 % hadof the cases were associated with pneumonia or bronchitis. However, only one case of Bordetella bronchiseptica has been reported in South Korea, and it was associated with peritonitis. In the current study, we report a case of Bordetella bronchiseptica pneumonia diagnosed in an immunocompromised patient.

Stimulatory effects of Bordetella bronchiseptica antigen on bone marrow cells and immune memory responses / 대한수의학회지

Bone marrow is a hematological and immunological organ that provides multiple immune cells, including B lymphocytes, and thus plays a critical role in the efficacy of vaccine. We previously demonstrated that Bordetella (B.) bronchiseptica antigen has high immunogenicity in spleen cells, a peripheral immune organ. In this study, we investigated the immunogenicity of B. bronchiseptica antigen in bone marrow cells, a central immune organ. B. bronchiseptica antigen increased the cellular activity of bone marrow cells and significantly enhanced the production of nitric oxide, IL-6, and TNF-alpha. Bone marrow cells primed with B. bronchiseptica antigen in vivo were harvested and stimulated with the same antigen in vitro. The stimulation of B. bronchiseptica antigen significantly increased the cellular activity and proliferation rate of the primed cells. B. bronchiseptica antigen also greatly induced the production of antigen-specific antibody in the primed cells. Taken together, the present study demonstrated that B. bronchiseptica antigen can stimulate bone marrow cells, a central immune organ, and recall the immune response of the primed bone marrow cells.

Evaluation of the immunogenicity of Bordetella bronchiseptica, a vaccine antigen / 대한수의학회지

Bordetella (B.) bronchiseptica is a causative agent of swine atrophic rhinitis that promotes colonization of the mucous membrane of the swine nasal cavity by Pasteurella (P.) multocida. Mixed infection with B. bronchiseptica and P. multocida leads to growth inhibition of pigs, resulting in significant economic loss. There are many commercial vaccines for atrophic rhinitis, including B. bronchiseptica as a killed vaccine antigen (Ag). However, the immunogenicity of killed B. bronchiseptica Ag has not yet been elucidated; therefore, this study was conducted to investigate the immunogenicity of killed B. bronchiseptica Ag and the type of immune response it induces. In vitro assays using mouse spleen cells and flow cytometry revealed that B. bronchiseptica Ag induced high proliferation capability of lymphocytes, especially B lymphocytes, and the proliferating cells showed a significant response to interleukin (IL)-2. B. bronchiseptica Ag also enhanced the production of IL-12, a representative cytokine for cell-mediated immunity. In vivo experiments using mice showed that the injection of B. bronchiseptica Ag markedly induced Ag-specific antibody. Taken together, these results indicate that B. bronchiseptica Ag has high immunogenicity by itself.

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.

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.

Analysis of microbial diversity of nitrifying bacteria by terminal restriction fragment length polymorphism / 生物工程学报

We analyzed the microbial diversity and quantity of nitrifying bacteria in the enrichment reactor by Terminal Restriction Fragment Length Polymorphism (T_RFLP), a cultured-independent molecular technique. The result indicated that nitrobacteria enriched the best, and the diversity index decreased 62.80% compared with the initial data. Nitrobacteria were predominant in the reactor. Meanwhile, we studied the microbial diversity before and after adding Nitrobacteria into shrimp ponds, and analyzed several major bacterial species that existed stably in the pond. According to the analysis by T_RFLP program, species including Brevibacillus brevis, Microbacterium lactium, Azoarcus indigens and Bordetella holmesii were the dominant bacteria in the ponds.

The Clinical Significance of Nasopharyngeal Carriages in Immunocompromised Children as Assessed / 대한혈액학회지

BACKGROUND: The aim of this study was to determine the prevalence of asymptomatic nasopharyngeal carriages in immunocompromized children by using a multiplex reverse transcriptase-polymerase chain reaction (mRT-PCR) assay kit. METHODS: We obtained clinical samples by nasopharyngeal swabs from 42 patients with underlying immune deficiency from May 20, 2008 to May 22, 2008. The children were free from signs of respiratory tract infections at the time of sampling. Isolated cDNA was extracted and after this the DNA was examined using a multiplex primer set for pneumonial bacteria detection (Seeplex(R) PneumoBacter ACE Detection, Seegene, Seoul, Korea). The amplified PCR products were separated on 2% agarose gels and stained with ethidium bromide and a screentape system (Lab901, Scottland, UK) and then they were compared. The nasopharyngeal swab culture was done simultaneously and this was compared with the results of mRT-PCR. RESULTS: A total of 42 patients (males: 24, females: 18) aged between 1.2 and 16.3 years (median: 9.2 years) were included in this study. The mRT-PCR detected bacteria (Streptococcus pneumoniae, Haemophilus influenzae, Chlamydia pneumoniae, and Bordetella pertussis) in 28 patients (66.6%). Of these 28 patients, 4 patients (14.3%) showed more than 2 bacteria: 2 patients were positive for two bacteria (S. pneumoniae and H. influenzae, H.influenzae and B. pertussis) and 2 patients were positive for three bacteria (S. pneumoniae, B. pertussis and C. pneumoniae, C. Pneumoniae, H. influenzae and B. pertussis). S. pneumoniae was cultured in one patient (2.4%). Conclusions: The mRT-PCR is a sensitive tool for the detection of the asymptomatic nasopharyngeal carriages. The clinical significance of the bacteria detected in immunocompromized patients by mRT-PCR will need further evaluation.

Detection of nasopharyngeal carriages in children by multiplex reverse transcriptase-polymerase chain reaction / 소아과

PURPOSE: The aim of this study was to determine the prevalence of asymptomatic nasopharyngeal carriages in children using a multiplex reverse transcriptase-polymerase chain reaction (mRT-PCR) assay kit. METHODS: We obtained nasopharyngeal swabs from 33 children without any underlying disease from July 25 to July 28, 2008. The children were free from the signs of respiratory tract infections at the time of sampling. DNA was extracted from the swabs and subjected to multiplex RT-PCR using a primer set for the detection of pneumococci (Seeplex(R) PneumoBacter ACE Detection Seegene, Seoul, Korea). The amplified PCR products were separated on 2% agarose gels and stained with either ethidium bromide or screen tape system (Lab901 Scotland, UK). RESULTS: A total of 33 children (male, 15 female, 18) aged between 3.2 and 16.3 (median, 8.2) years were included in this study. The mRT-PCR detected colonized bacteria (Streptococcus pneumoniae, Hemophilus influenzae, Chlamydia pneumoniae, and Bordetella pertussis) in 30 children (90.9%). Of these, 13 children (39.4%) showed more than 2 bacteria: 12 children were positive for 2 bacteria (S. pneumoniae and H. influenzae) and 1 child was positive for 3 bacteria (S. pneumoniae, H. influenzae, and C. pneumoniae). CONCLUSION: mRT-PCR was found to be a sensitive tool for the detection of asymptomatic nasopharyngeal carriages. Clinical significances of the bacteria detected by mRT-PCR will have to be evaluated in the future.

Discriminative PCR of Bordetella pertussis from closely related Bordetella species using 16S rDNA Gene

BACKGROUND: Polymerase-chain reaction (PCR) detection is useful to diagnosis of pertussis at initial stage because the growth rate of Bordetella pertussis (B. pertussis) is relatively slow. Currently, the primer set for the insertion sequence IS481 (BP primer) is used widely for PCR detection of B. pertussis. However, the cross-reactivity of BP primer set with Bordetella holmesii (B. holmesii) was reported recently. Therefore, discrimination of B. pertussis and B. holmesii is needed in PCR step. For this reason, we developed new primer sets based on 16S rDNA sequence for diagnostic use and estimated the efficiency of these new primer sets. MATERIALS AND METHODS: The specific PCR primers were designed from the aligned sequence matrix of 16S rDNA genes of various Bordetella species. The specificity of designed primers were estimated using clinically important 4 Bordetella species, B. pertussis, B. holmesii, Bordetella parapertussis (B. parapertussis) and Bordetella bronchiseptica (B. bronchiseptica). The sensitivity to B. pertussis of designed primers was also estimated and compared with BP primer set. RESULTS: As the results, the developed new primer set successfully distinguished B. pertussis and other Bordetella species containing B. holmesii. In the sensitivity assay, the detectable limits of 16S-F2/16S-R1 primer set for B. pertussis were revealed as 5 pg of genomic DNA and 105 cells/mL of cell suspension. In addition to these, identical results between BP with primer and new primer were obtained in clinical samples. CONCLUSION: In this study, the specific primer set for B. pertussis was developed based on 16S rDNA sequence and this primer set did not show cross-reactivity to B. holmesii. In addition to these, the applicability of this primer set to the clinical specimens was also confirmed.

Discriminative PCR of Bordetella pertussis from closely related Bordetella species using 16S rDNA Gene

BACKGROUND: Polymerase-chain reaction (PCR) detection is useful to diagnosis of pertussis at initial stage because the growth rate of Bordetella pertussis (B. pertussis) is relatively slow. Currently, the primer set for the insertion sequence IS481 (BP primer) is used widely for PCR detection of B. pertussis. However, the cross-reactivity of BP primer set with Bordetella holmesii (B. holmesii) was reported recently. Therefore, discrimination of B. pertussis and B. holmesii is needed in PCR step. For this reason, we developed new primer sets based on 16S rDNA sequence for diagnostic use and estimated the efficiency of these new primer sets. MATERIALS AND METHODS: The specific PCR primers were designed from the aligned sequence matrix of 16S rDNA genes of various Bordetella species. The specificity of designed primers were estimated using clinically important 4 Bordetella species, B. pertussis, B. holmesii, Bordetella parapertussis (B. parapertussis) and Bordetella bronchiseptica (B. bronchiseptica). The sensitivity to B. pertussis of designed primers was also estimated and compared with BP primer set. RESULTS: As the results, the developed new primer set successfully distinguished B. pertussis and other Bordetella species containing B. holmesii. In the sensitivity assay, the detectable limits of 16S-F2/16S-R1 primer set for B. pertussis were revealed as 5 pg of genomic DNA and 105 cells/mL of cell suspension. In addition to these, identical results between BP with primer and new primer were obtained in clinical samples. CONCLUSION: In this study, the specific primer set for B. pertussis was developed based on 16S rDNA sequence and this primer set did not show cross-reactivity to B. holmesii. In addition to these, the applicability of this primer set to the clinical specimens was also confirmed.

Effect of Casing Layer on Growth Promotion of the Edible Mushroom Pleurotus ostreatus

Various bacteria were isolated from the casing layer soil of the culture bed of P. ostreatus and their role in fruiting body induction of the edible mushroom, P. ostreatus, was investigated. Analysis of the bacterial community isolated from the casing layer soil revealed that the composition of genera and number of cultivable bacteria were different for each sterilizing treatment. Bordetella was predominant in the bulk soil whereas Flavobacterium was predominant after sterilization of the casing layer soil. Fluorescent Pseudomonas was predominant in the non-sterilized casing layer soil. Total number of the bacterial genera in the casing layer soil was higher than that in the bulk soil. In particular, an increase in the fluorescent Pseudomonas population was observed in the non-sterilized casing layer accompanied by induction of fruiting body and enhanced mushroom production yield. The results suggested that specific bacterial populations in the casing layer play an important role in the formation of primodia and the development of basidiome in P. ostreatus.

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...

Outbreak of pertussis in Basra, Iraq

A total of 133 pertussis cases were studied during an outbreak in Basra from June to December 1996. Most were females and were immunized. Bordetella spp. was isolated in 48.1% of the cases. The isolation rate was highest among infants and decreased with increasing age, and was highest during the catarrhal stage. B. pertussis was the most common species; however, B. parapertussis infection did occur. There were some severe cases of pertussis among infants caused mainly by B. pertussis and dual Bordetella infection. Infection was transmitted by close contact with a pertussis case

Evaluation of serum opsonic capacity against Bordetella pertussis by chemiluminescence

Serum opsonic capacity against B, pertussis was studied by using quantitative chemiluminescence [CL], a method known to have several advantages over conventional methods in evaluating opsonization and phagocytosis. Sera from unvaccinated infants was shown not to contain opsonins against B. pertussis and in unvaccinated infants suffering from whooping cough, no opsonins were detected. In adults fully vaccinated during childhood, antibody liters decreased with time. Therefore, antibody transfer to infants is negligible. The CL assay is simple, rapid, and reproducible, offering new possibilities to evaluate humoral immune mechanisms and phagocytosis in whooping cough

Diagnosis of Bordetella Pertussis Infections by Polymerase Chain Reaction

PURPOSE: Pertussis, a respiratory tract infection caused by Bordetella pertussis, is an important cause of morbidity in children. But diagnosis of pertussis is often delayed because of late development of typical symptoms and difficulties in culture. There has been no bacteriologically confirmed case of B. pertussis infection in Korea. Lower respiratoy tract may be involved in pertussis. We performed the polymerase chain reaction(PCR) assay for B. pertussis from children with lower respiratory tract infections. METHODS: One hundred eighty nine nasopharyngeal aspirates were collected from children with lower respiratory tract infections during the period from November 1990 to February 1995. Three 24-mer primers derived from DNA sequences upstream of the structural genes for the porin proteins of Bordetella: P1 is shared by all three species and P2 is specific for B. pertussis and P3 is specific for B. parapertussis and B. bronchiseptica. Amplifications resulted in 159-bp PCR products specific for B. pertussis and 121-bp PCR products specific for B. parapertussis and B. bronchiseptica. A confirmatory cleavage of 159-bp PCR products of B. pertussis by Hae III revealed two bands of 22 and 137 bp. RESULTS: B. pertussis specific PCR products were visualized in 6 patients during 1991 and 5 of these had received appropriate doses of the combined DPT vaccine. They had had cough over 2 weeks in all and high fever in 4. They had been diagnosed as viral pneumonia in 5 and bronchiolitis in 1, but viral cultures for respiratory syncytial virus, parainfluenza virus, influenza virus, adenovirus were negative. There was no PCR product compatible with B. parapertussis or B. bronchiseptica. CONCLUSIONS: PCR assay is effective in diagnosis of B. pertussis infections. We suggest that there was an epidemic of pertussis in 1991 despite high rate of pertussis vaccine coverage in Korea.