Clinical repercussions of Glanders (Burkholderia mallei infection) in a Brazilian child: a case report

Abstract Glanders is a relatively unknown zoonotic disease caused by Burkholderia mallei. This bacterium affect solipeds and humans, and can be used as a biological warfare. Glanders is characterized as an occupational disease. We report the case of an 11-year-old boy who was presented to an emergency department with chest pain and dyspnea. He evolved into septic shock, pneumonia, and multiple abscesses. B. mallei was found in the exudate culture. Human infection is rare and difficult to confirm. The knowledge on glanders is important for differential diagnosis from other serious illnesses causing pneumonia and multiple abscesses.

Produção e purificação parcial de PPD-maleína para diagnóstico do mormo em equídeos / Partially purified malleo-proteins production for glanders diagnosis in Equidae

Objetivou-se com este estudo produzir e purificar parcialmente a PPD-maleína a partir de amostras de Burkholderia mallei isoladas de equídeos no Brasil com potencial para uso no diagnóstico do mormo. As linhagens de B. mallei fenotipicamente caracterizadas e de virulência comprovada foram inoculadas em caldo Dorset-Henley para crescer e metabolizar. Em seguida, as proteínas foram separadas por precipitação com ácido tricloroacético e precipitadas com sulfato de amônia. As PPDs-maleínas foram concentradas em 1,0mg/mL e na avaliação realizada em cobaios foi eficaz no desenvolvimento da hipersensibilidade do tipo tardia e consequentemente na identificação de animais verdadeiro positivos e exclusão dos verdadeiro negativos, sendo uma possibilidade em potencial para utilização no diagnóstico do mormo.

The objective of this study was to produce and partially purify Malleo-protein from Burkholderia mallei samples isolates from Equidae in Brazil with potential for use in the diagnosis of glanders. The strain B. mallei phenotypically characterized and proven virulent was inoculated into broth Dorset-Henley to grow and metabolize. The proteins were separated by trichloroacetic acid precipitation and amonium sulfate precipitation. The PPD mallein was concentrated 1.0mg/mL and biologically tested in guinea pigs. It was effective in the development of delayed-type hypersensitivity and consequently to identify true-positive animals and to exclude of true negatives. There is the possibility for potential use in the glanders diagnosis in Equidae.

Assessment of the effectiveness of the PPD-mallein produced in Brazil for diagnosing glanders in mules

To assess the potency of the PPD-mallein produced in Brazil, five animals were from a property identified as a focus of glanders. These animals had suggestive clinical signs of the disease and the other five, from a property free from glanders, showed no clinical signs and were serology negative (control group). PPD-mallein from Burkholderia mallei was obtained by precipitation with trichlo-roa-cetic acid and ammonium sulfate. The animals were inoculated according to the criteria established by Department of Agriculture, Livestock and Supply (MAPA) for the diagnosis of glanders. After 48 h of application of PPD-mallein, there was swelling in the area of application, presence of ocular secretion and tears in sick animals. The control group showed no inflammatory reaction at the site of inoculation of PPD-mallein. This immunogen produced in Brazil and still being tested was effective for identifying the infection in true positive animals and excluding the truly negative ones, being a new possibility for diagnosis and control of glanders.

Glanders in donkeys (Equus asinus) in the state of Pernambuco, Brazil: a case report

The clinical, anatomical and histopatological findings of glanders diagnosis in donkeys in the state of Pernambuco-Brazil are reported. The animals were euthanized and necropsied, and evaluated for lesions in respiratory and lymphatic systems, confirming the disease by isolation of Burkholderia mallei and Strauss test.

Caracterização fenotípica e molecular de amostras de Burkholderia mallei isoladas na Região Nordeste do Brasil / Phenotypic and molecular characterization of Burkholderia mallei isolated in northeastern Brazil

Objetivou-se com este trabalho realizar o estudo bioquímico e molecular de amostras de Burkholderia mallei isoladas de eqüídeos com diagnóstico clínico e sorológico para o mormo e provenientes da Região Metropolitana do Recife-PE e Zona da Mata dos Estados de Alagoas e Pernambuco. Foram realizadas as técnicas microbiológicas para o isolamento e identificação fenotípica de B. mallei e as técnicas moleculares de ribotipagem-PCR e RAPD-PCR. Das oito amostras estudadas, quatro apresentaram pequenas variações fenotípicas. Nas técnicas moleculares, as amostras formaram quatro grupos de diferentes perfis ribotípicos, demonstrando também quatro perfis genotípicos. Houve associação nos resultados da Ribotipagem-PCR e RAPD-PCR. As variações nos perfis ribotípicos e genotípicos foram associadas às diferentes regiões estudadas. De acordo com os resultados obtidos, conclui-se que as pequenas variações bioquímicas não estão associadas aos diferentes perfis moleculares e que essas diferenças demonstram uma heterogeneidade que está associada à procedência das amostras, indicando que a infecção nos animais ocorre por clones diferentes das amostras analisadas.

The objective of this paper was to study the molecular performance and phenotypic characterization of Burkholderia mallei isolated from horses with clinical and serological diagnosis of glanders, originating from the Metropolitan District of Recife and Zona da Mata of Pernambuco and Alagoas. The isolation and biochemical identification of B. mallei was carried out by microbiological and molecular techniques of PCR-fingerprinting and RAPD-PCR. From the eight samples studied, four showed little phenotype variations. In the molecular tests, the samples formed 4 groups of different ribotype profiles and 4 genotype profiles. There was some association of PCR-fingerprinting with RAPD-PCR results. It was concluded that the slight biochemical variations were not associated with different molecular profiles. They also indicated that these differences show heterogeneity associated with the origin of the sample, indicating that the infection was caused by clones of different strains and that the polymorphism of DNA observed could make it difficult to choose one standard strain for an immune prophylactic treatment of glanders.

Proteinograma sérico em muares naturalmente infectados pela Burkholderia mallei / Serum proteinogram in mules naturally infected by the Burkholderia mallei

The Burkholderia mallei is the causing bacterium of glanders, illness of high morbidity and lethality for the equides, and also one zoonosis. Recently diagnostic in the Pernambuco and Alagoas States/Brazil, it comes decimating populations of equides in the Zone of Forest Region of these states, causing great damages to the sugar cane plantation activity that it uses such animals as traction. The objective this work was to investigate the proteins alterations caused by glanders. Had been studied ninety adult mules, of different races, destined to the work, proceeding from the sugar cane plantation region, Zone of Forest, in the Pernambuco State/Brazil. These were divided in three groups: G1: thirty animals with negative serology for glanders; G2: thirty animals with positive serology for glanders and without apparent symptoms and G3: thirty animals with positive serology for glanders and with apparent symptoms. The gotten results, referring the medium of the parameters studied respectively for G1, G2 and G3 had been: total serum protein 7.33; 7.73 and 7.46g/dl; albumin 2.57; 2.43 and 1.81g/dl; globulins 4.37; 4.86 and 5.64g/dl; albumin/ globulins ratio 0.55; 0.47 and 0.34g/dl; alpha-globulin 1.06; 1.33 and 1.33g/dl; beta-globulin 1.10; 1.21 and 1.80g/dl and gamma-globulin 2.21; 2.32 and 2.51g/dl. It is concluded that the variations for the studied parameters were significant; the increase of the globulins characterizes an antigenic stimulus in the sick animals, as well as an inversion in the albumin/globulins ratio for the animals with apparent clinic in relation to the excessively animal ones. These findings could be considered in the diagnosis, prognostic and in future research that they aim at to study forms of immunization against this illness.

A Burkholderia mallei é a bactéria causadora do mormo, doença de alta morbidade e letalidade para os eqüídeos, e também uma zoonose. Recentemente diagnosticada nos estados de Pernambuco e Alagoas, vem dizimando populações de eqüídeos na Região da Zona da Mata destes estados, causando grandes prejuízos à atividade canavieira que utiliza tais animais como tração. Objetivou-se com este trabalho investigar as alterações protéicas causadas pelo mormo. Foram estudados 90 muares adultos, de diferentes raças, destinados ao trabalho, provenientes da região canavieira, Zona da Mata, do Estado de Pernambuco. Estes foram divididos em três grupos: G1: composto por trinta animais sorologicamente negativos para o mormo; G2: composto por trinta animais sorologicamente positivos e sem sintomatologia clínica aparente e G3: composto por trinta animais sorologicamente positivos e com sintomatologia clínica aparente. Os resultados obtidos, referentes à média dos parâmetros estudados para G1, G2 e G3 foram respectivamente: proteína sérica total 7,33; 7,73 e 7,46g/dl; albumina 2,57; 2,43 e 1,81g/dl; globulinas 4,37; 4,86 e 5,64g/dl; relação albumina/globulinas 0,55; 0,47 e 0,34g/dl; alfaglobulina 1.06; 1.33 e 1,33g/dl; beta-globulina 1,10; 1,21 e 1,80g/dl e gama-globulina 2,21; 2,32 e 2,51g/dl. Conclui-se que as variações para os parâmetros estudados foram significativas, o aumento das globulinas caracteriza um estímulo antigênico nos animais positivos, bem como uma inversão na relação albumina/globulinas para os animais com clínica aparente em relação aos demais animais. Estes achados poderão ser considerados no diagnóstico, prognóstico e em pesquisas futuras que visem estudar formas de imunização contra esta importante enfermidade.

Mormo em eqüídeos nos Estados de Pernambuco e Alagoas / Glanders in horses and mules of the states of Pernambuco and Alagoas, Brazil

Com base em aspectos clínico-patológicos, epidemiológicos e sorológicos, bem como pelo isolamento de Burkholderia mallei, diagnosticaram-se focos de mormo em eqüídeos na Zona da Mata dos Estados de Pernambuco e Alagoas. Clinicamente observaram-se hipertermia, respiração ruidosa, inapetência, emagrecimento progressivo até caquexia, congestão nasal e descarga mucopurulenta, erosões, úlceras e cicatrizes na mucosa nasal. Os linfonodos mandibulares e cervicais superficiais apresentavam-se aumentados de volume, com abscessos e fístulas exsudando material purulento. Na pele, ao longo dos vasos linfáticos, havia formação de nódulos firmes e abscessos que, nos casos crônicos, fistulavam e ulceravam deixando cicatrizes estrelares. A histopatologia dos nódulos de diversos sítios, tanto dos eqüídeos como dos cobaios utilizados na prova de Straus, revelou lesões granulomatosas e piogranulomas. Os dados deste estudo evidenciam a emergência da doença nesta área e permitem recomendar inquéritos soro-epidemiológicos nos Estados de ocorrência dos surtos e nos Estados limítrofes.

Based on clinical-pathological, epidemiological and serological aspects, as well as by the isolation of Burkholderia mallei, foci of glanders were diagnosed in equids of the "Zona da Mata" in the states of Pernambuco and Alagoas. Clinically there was hyperthermia, noisy respiration, loss of appetite, progressive loss of weight leading to emaciation, nasal congestion and muco-purulent discharge, erosions, ulcers and scars of the nasal mucosa. The size of submaxillary and cervical lymphonodes was increased, which contained abscesses and fistules exudating purulent material. In the skin, accompanying the lymph vessels, there were firm nodules and abscesses which in chronic cases fistulated and ulcerated leaving stellar scars. The histopathology of the nodules at various sites, from the horses as well as from the guinea-pigs used for the Strauss test, showed granulomatous lesions and pyogranulomas. The results of this study showed that glanders in the region was emerging and further sero-epidemiological studies are urgent for the eradication of the disease.