Detection of Mycoplasma capricolum subsp. capripneumoniae and Mannheimia haemolytica as causative agents of pleuropneumonia in goats / Detección de Mycoplasma capricolum subsp. capripneumoniae y Mannheimia haemolytica como agentes causales de pleuro-neumonia en cabras

Abstract Objective: The aim of the study was detection of two major causative agents of pleuropneumonia, Mycoplasma capricolum subsp. capripneumoniae (Mccp) and Mannheimia haemolytica, in goats. To the best of our knowledge, this study is the first investigation of Mccp in Iran. Methods: 50 grossly suspected lungs to pleuropneumonia and 10 healthy samples were collected from Shiraz abattoir. Results: Histopathological evaluation of tissue samples showed various diagnosed pneumonias including 40% bronchointerstitial pneumonia (20 samples), 34% interstitial pneumonia (17 samples), 10% fibrinopurulent bronchopneumonia (5 samples), 12% purulent bronchopneumonia (6 samples) and 4% chronic pneumonia (2 samples). In molecular study, all 50 suspected samples and 10 healthy ones by PCR showed no Mccp positive sample, but the detection rate of M. haemolytica in suspected samples was 14% and in healthy lungs was zero. Conclusions: It may be concluded that goats referred to Shiraz abattoir is free of Mccp. Further sampling and molecular testing at the level of suspected herds to CCPP can be useful.

Resumen Objetivo: El objetivo del estudio fue la detección de dos agentes causantes principales de pleuroneumonía, Mycoplasma capricolum subsp. Capripneumoniae (Mccp) y Mannheimia haemolytica, en cabras. Hasta donde sabemos, este estudio es la primera investigación de Mccp en Irán. Métodos: 50 pulmones muy sospechosos de pleuroneumonía y 10 muestras sanas se obtuvieron del matadero de Shiraz. Resultados: La evaluación histopatológica de muestras de tejido mostró varias neumonías diagnosticadas, incluyendo 40% de neumonía broncointersticial (20 muestras), 34% de neumonía intersticial (17 muestras), 10% de bronconeumonía fibrinopurulenta (5 muestras), 12% de bronconeumonía purulenta (6 muestras) y 4% neumonía crónica (2 muestras). En un estudio molecular, las 50 muestras sospechosas y 10 sanas por PCR no mostraron una muestra positiva de Mccp, pero la tasa de detección de M. haemolytica en muestras sospechosas fue del 14% y en pulmones sanos fue cero. Conclusiones: se puede concluir que las cabras referidas al matadero Shiraz están libres de Mccp. La realización de muestreo adicional y pruebas moleculares a nivel de rebaños sospechosos para CCPP puede ser útil.

Glomerulonefritis infecciosa y pleuroneumonía por Streptococcus pneumoniae. Caso clínico pediátrico / Infectious glomerulonephritis and pleuropneumonia due to Streptococcus pneumoniae. Pediatric clinical case

La glomerulonefritis aguda desencadenada por Streptococcus pneumoniae es una patología de baja prevalencia. Existen diversos reportes que comunican distintas cepas nefritogénicas; sin embargo, la 6C ha sido escasamente señalada como tal.Se presenta el caso de un paciente de 4 años, quien ingresó a Terapia Intensiva con pleuroneumonía por Streptococcus pneumoniae serotipo 6C y desarrolló, de modo concomitante, edemas, hipertensión arterial, hematuria, proteinuria, disminución del filtrado glomerular y del nivel de complemento C3. Se diagnosticó glomerulonefritis aguda. Su evolución fue satisfactoria en un breve plazo. Esta patología, por lo general, es de curso transitorio y benigno; sin embargo, en ocasiones, puede complicar la evolución de un paciente críticamente enfermo, por lo cual se hace necesario tenerla entre los diagnósticos diferenciales para considerar.

Acute glomerulonephritis caused by Streptococcuspneumoniaeis a low prevalence pathology. There are several reports communicating different nephritogenic serotypes, however, 6C has been scarcely indicated as such. It is presented the case of a 4-year-old patient who entered Intensive Therapy Unit with pleuropneumonia due to Streptococcuspneumoniae serotype 6C and concomitantly developed edemas, arterial hypertension, hematuria, proteinuria, decreased glomerular filtration rate and C3 complement level. Acute glomerulonephritis was diagnosed. His evolution was satisfactory in a short time. This pathology is usually of a transitory and benign course; however, sometimes it can potentially complicate the evolution of a critically ill patient, so it is necessary to have it among the differential diagnoses to consider.

In silico analysis of putative drug and vaccine targets of the metabolic pathways of Actinobacillus pleuropneumoniae using a subtractive/comparative genomics approach

Actinobacillus pleuropneumoniae is a Gram-negative bacterium that resides in the respiratory tract of pigs and causes porcine respiratory disease complex, which leads to significant losses in the pig industry worldwide. The incidence of drug resistance in this bacterium is increasing; thus, identifying new protein/gene targets for drug and vaccine development is critical. In this study, we used an in silico approach, utilizing several databases including the Kyoto Encyclopedia of Genes and Genomes (KEGG), the Database of Essential Genes (DEG), DrugBank, and Swiss-Prot to identify non-homologous essential genes and prioritize these proteins for their druggability. The results showed 20 metabolic pathways that were unique and contained 273 non-homologous proteins, of which 122 were essential. Of the 122 essential proteins, there were 95 cytoplasmic proteins and 11 transmembrane proteins, which are potentially suitable for drug and vaccine targets, respectively. Among these, 25 had at least one hit in DrugBank, and three had similarity to metabolic proteins from Mycoplasma hyopneumoniae, another pathogen causing porcine respiratory disease complex; thus, they could serve as common therapeutic targets. In conclusion, we identified glyoxylate and dicarboxylate pathways as potential targets for antimicrobial therapy and tetra-acyldisaccharide 4′-kinase and 3-deoxy-D-manno-octulosonic-acid transferase as vaccine candidates against A. pleuropneumoniae.

Pasteurella multocida type A as the primary agent of pneumonia and septicaemia in pigs / Pasteurella multocida tipo A como agente primário de pneumonia e septicemia em suínos

In order to understand better the pathological aspects and spread of Pasteurella multocida type A as the primary cause of pneumonia in pigs, was made an experiment with intranasal inoculation of different concentrations of inocula [Group (G1): 108 Colony Forming Units (CFU)/ml; G2: 107 CFU/ml; G3: 106 CFU/ml and G4: 105 CFU/ml], using two pigs per group. The pigs were obtained from a high health status herd. Pigs were monitored clinically for 4 days and subsequently necropsied. All pigs had clinical signs and lesions associated with respiratory disease. Dyspnoea and hyperthermia were the main clinical signs observed. Suppurative cranioventral bronchopneumonia, in some cases associated with necrosuppurative pleuropneumonia, fibrinous pericarditis and pleuritic, were the most frequent types of lesion found. The disease evolved with septicaemia, characterized by septic infarctions in the liver and spleen, with the detection of P. multocida type A. In this study, P. multocida type A strain #11246 was the primary agent of fibrinous pleuritis and suppurative cranioventral bronchopneumonia, pericarditis and septicaemia in the pigs. All concentrations of inoculum used (105-108 CFU/ml) were able to produce clinical and pathological changes of pneumonia, pleuritis, pericarditis and septicemia in challenged animals...

Para entender melhor os aspectos patológicos e disseminação de Pasteurella multocida tipo A como causa primária de pneumonia em suínos foi realizado um experimento com inoculação intranasal de diferentes concentrações de inóculos [Grupo (G1): 108 Unidades Formadoras de Colônias (UFC)/ml; G2: 107 UFC/ml; G3: 106 UFC/ml e G4: 105 UFC/ml], usando dois suínos por grupo. Esses suínos foram obtidos de um rebanho de alto status sanitário. Os animais foram monitorados clinicamente por quarto dias e subsequentemente necropsiados. Todos os suínos apresentaram sinais clínicos e lesões associadas com doença respiratória. Dispneia e hipertermia foram os principais sinais clínicos observados. Broncopneumonia cranioventral supurativa, em alguns casos associados com pleuropneumonia necrossupurativa, pleurites e pericardite fibrinosa foram mais frequentes. A doença evoluiu com septicemia, caracterizada por infartos sépticos no fígado e baço, com detecção de P. multocida. Neste estudo, P. multocida tipo A isolado 11246 foi agente primário de pleurite fibrinosa e broncopneumonia cranioventral supurativa, pericardite fibrinosa e septicemia em suínos. Todas as concentrações de inóculo utilizado (105-108 UFC/ml) foram capazes de produzir sinais clínicos e patológicos de alterações de pneumonia, pleurites, pericardites e septicemia nos animais...

Prevalence and Characterization of Actinobacillus pleuropneumoniae Isolated from Korean Pigs

Actinobacillus pleuropneumoniae causes porcine pleuropneumoniae which is one of severe threats to the swine industry. In total, 54 strains of Actinobacillus pleuropneumoniae were isolated from 443 pigs between 2012 and 2013 in Korea. Isolates were classified into serotypes 1, 2, 5, 7, 12, and unclassified by multiplex PCR. Genotypes of isolates were divided into three groups according to the sequence of the omlA gene. The antimicrobial resistance rate of serotype 1 was slightly higher than that of serotype 5. In conclusion, to block and treat porcine pleuropneumonia, it is necessary to conduct ongoing characterization of A. pleuropneumoniae isolated from pigs.

Doenças de ovinos diagnosticadas no Laboratório de Anatomia Patológica Animal da Universidade Federal de Mato Grosso do Sul (1996-2010) / Sheep diseases diagnosed at the Laboratory of Animal Pathology, Federal University of Mato Grosso do Sul, Brazil (1996-2010)

Nas últimas décadas a pecuária ovina cresceu significativamente no Brasil. Concomitantemente, grupos de pesquisas e laboratórios de diagnósticos realizam estudos retrospectivos com a finalidade de fornecer subsídios técnico-científicos para os médicos veterinários. Desta forma, realizou-se um estudo de prevalência nos arquivos do Laboratório de Anatomia Patológica Animal (LAP) da Universidade Federal de Mato Grosso do Sul (UFMS) no período de Janeiro de 1996 a Dezembro de 2010. O Laboratório de Bacteriologia da UFMS e o Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul forneceram apoio diagnóstico nos casos de mannheimiose pulmonar e scrapie, respectivamente. Os laudos da espécie ovina foram revisados e agrupados em conclusivos e inconclusivos, dos quais foram excluídos os casos experimentais e de outros estados e países. Os casos conclusivos foram classificados de acordo com a etiologia da doença. Os exames da espécie ovina somaram 331 laudos (3,97 %) de um total de 8.333 casos diagnosticados no período. Destes, foram excluídos sessenta e quatro (19,3%) casos experimentais e materiais oriundos de outros estados ou países. Dos 267 casos remanescentes, 87 (32,6%) foram inconclusivos e 180 (67,4%) considerados conclusivos, sendo 60 (33,3%) doenças infecciosas e parasitárias; 45 (25%) intoxicações e toxi-infecções; 41 (22,8%) "lesões sem causa definida"; 22 (12,2%) doenças metabólicas e nutricionais; 10 (5,6%) foram classificadas como "outros distúrbios" e 2 (1,1%) neoplasmas. A hemoncose, intoxicação por Brachiaria spp., pleuropneumonias, broncopneumonias, pneumonias fibrinonecrosante ou fibrinossupurativa sem causa definida e a intoxicação por cobre foram as doenças mais prevalentes no período estudado. Dois casos de scrapie foram diagnosticados no período.

Sheep farming has increased significantly in Brazil during the last decades. Concurrently, research groups and diagnostic laboratories compile data and perform retrospective studies to provide important insight for professionals. A prevalence study from January 1996 to December 2010 was carried out in the archives of Laboratório de Anatomia Patológica Animal (LAP), Universidade Federal de Mato Grosso do Sul (UFMS). Laboratório de Bacteriologia, UFMS, and Setor de Patologia Veterinária at Universidade Federal do Rio Grande do Sul helped on the diagnostic of pulmonary mannheimiosis and scrapie respectively. The reports for sheep were reviewed and grouped into conclusive and inconclusive ones. The conclusive cases were classified according to the etiology of the disease. In the period, 331 exams (3.97%) were done. Sixty-four experimental cases and materials from other states or countries (19.3%) were excluded. Remaining cases (267), eighty-seven (32.6%) were inconclusive and 180 (67.4%) were considered conclusive reports, were classified according to the etiology: 60 (33.3%) infectious and parasitary diseases; 45 (25%) were poisonings and toxi-infections; 41 (22.8%) were summarized as "injuries without apparent cause"; 22 (12.2%) cases of metabolic and nutritional diseases; 10 (5.6%) were classified as "other disorders" and 2 (1.1%) case of neoplasms. Haemonchosis, fibrinonecrotic or fibrinopurulent pleuropneumonia, bronchopneumonia and pneumonia, poisonings by Brachiaria spp. and copper poisoning were the most prevalent diseases in sheep. Two cases of scrapie have been diagnosed in this period.

Serotype and antimicrobial susceptibility of Actinobacillus pleuropneumoniae isolates from pigs in Korea / 대한수의학회지

Actinobacillus (A.) pleuropneumoniae is the causative agent of pleuropneumonia which is one of the most important respiratory diseases in pigs worldwide. A total of 32 A. pleuropneumoniae isolates from diseased pigs during 2008 to 2010 were serotyped by polymerase chain reaction method. The susceptibility of the isolates to 13 antimicrobial agents were determined by disk diffusion test. In all the 32 isolates examined in this study, serotype 5 (16 isolates: 50%), 1 (7 isolates: 21.9%), 2 (5 isolates: 15.6%) and 12 (1 isolate: 3.1%) were found. Of all tested antimicrobial agents, resistance to oxytetracycline was found in 96.9% of isolates, followed by resistance to amikacin (81.2%), neomycin (68.7%), kanamycin (53.1%), penicillin (50.0%), gentamicin (43.7%), florfenicol (25.0%), ampicillin (18.7%), colistin (9.4%), trimethoprim/sulfamethoxazole, ceftiofur (8.3%), amoxicillin/clavulanic acid (3.1%) and enrofloxacin (0%). Oxytetracycline or florfenicol-resistant isolates were examined for the presence of resistance gene. Among the 31 oxytetracycline-resistant isolates, tetB, tetH and tetO genes were detected in 22 (71%), 8 (26%) and 1 (3%) isolates, respectively. The floR genes were detected in 8 (100%) of the 8 florfenicol-resistant A. pleuropneumoniae isolates.

Detecção direta de Actinobacillus pleuropneumoniae em órgãos de suídeos do estado de São Paulo pela técnica de reação em cadeia pela polimerase (nested-PCR) / Direct detection of Actinobacillus pleuropneumoniae in suidae organs in São Paulo state, Brazil, by polymerase chain reaction (nested-PCR)

A pleuropneumonia suína, causada pelo Actinobacillus pleuropneumoniae, é uma importante doença respiratória, responsável por prejuízos e queda de produtividade nas criações. Este trabalho teve como objetivo determinar a ocorrência de A. pleuropneumoniae em amostras de campo, mediante a adaptação e emprego de uma técnica de nested-PCR dirigida ao gene Apx IV. Definiu-se a sensibilidade analítica das técnicas de PCR e nested-PCR utilizando a amostra padrão A. pleuropneumoniae sorotipo III, em concentrações de DNA variando entre 30 µg/mL a 0,01 ng/ mL. Um total de trinta e sete amostras de campo encaminhadas ao Instituto Biológico entre 1995 a 2007 foram analisadas pelas técnicas de PCR e nested-PCR. A avaliação da sensibilidade analítica revelou que a PCR possui capacidade de gerar sinal a partir de 2 ng/mL de DNA extraído e a nested-PCR a partir de 0,4 ng/mL. Uma vez que a nested-PCR apresentou sensibilidade analítica cinco vezes maior se comparada à PCR para detecção de A. pleuropneumoniae em amostra padrão, o seu emprego pode minimizar a ocorrência de resultados tipo "falso-negativo". Dentre as amostras testadas, dez foram positivas à nested-PCR, sendo observada a ocorrência de A. pleuropneumoniae em nove diferentes animais, um deles javali. A presente técnica de nested-PCR pode ser utilizada para detecção direta de A. pleuropneumoniae em amostras de campo, mesmo após congelamento da amostra por longos períodos e sem necessidade de isolamento bacteriano prévio.

Porcine pleuropneumonia, caused by Actinobacillus pleuropneumoniae, is an important respiratory disease, responsible for economic losses and reduced productivity. The aim of this study was to determine occurrence of A. pleuropneumoniae in field samples, using an adapted nested-PCR reaction targeting the Apx IV gene. Different DNA concentrations (from 30 µg/mL to 0.01 ng/mL) of A. pleuropneumoniae serotype III reference strain were used to determine the level of sensitivity of first generation and nested-PCR reactions. Thirty-seven field samples sent to Instituto Biológico from 1995 to 2007 were tested by PCR and nested-PCR. Determination of the level of sensitivity showed that PCR could amplify to 2 ng/mL of extracted DNA and nested-PCR to 0.4 ng/mL. Since the nested reaction exhibited a level of sensitivity 5 times greater than the PCR reaction to detect a reference strain, using nested-PCR could minimize the occurrence of false-negative results. Among tested samples, 10 of them were nested-PCR positive, showing occurrence of A. pleuropneumoniae in 9 different animals (including one wild boar). This nested-PCR reaction can be used for direct detection of A. pleuropneumoniae in field samples, even after frozen storage for long periods, without the need for previous bacterial isolation.

Construction and characterization of Actinobacillus pleuropneumoniae serovar 7 live attenuated vaccine strain co-expressing ApxIA / 生物工程学报

Actinobacillus pleuropneumoniae (A. pleuropneumoniae), the causative agent of porcine contagious pleuropneumonia (PCP), is a significant pathogen of the world pig industry, vaccination is potentially an effective tool for the prevention of PCP. The purpose of present study was to enhance the immunogenicity of A. pleuropneumoniae live vaccine strain HB04C- (serovar 7), which was unable to express ApxIA, and to develop effective multivalent vaccines for the respiratory pathogens based on the attenuated A. pleuropneumoniae. We introduced a shuttle vector containing intact apxIA gene into HB04C-, generating HB04C2, an A. pleuropneumoniae serovar 7 live attenuated vaccine strain co-expressing ApxIA. Then we investigated the biological characteristics of HB04C2. We found that the shuttle vector expressing ApxIA was stable in HB04C2, and the growth ability of HB04C2 was not affected by the shuttle vector. We observed that HB04C2 elicited detectable antibodies against ApxIA and ApxIIA when it was administrated intratracheally as a live vaccine in pigs, and all immunized pigs were protected from heterologous virulent A. pleuropneumoniae (serovar 1) challenge. In conclusion, we demonstrated that A. pleuropneumoniae live vaccine could be used as a vector for expression of heterologous antigens.

Abscesos pleurales producidos por Staphylococcus aureus y Streptococcus spp. beta-hemolítico como secuela de pleuroneumonía en un caballo.Reporte de un caso / Abscessos pleurais causados por Staphylococcus aureus e Streptococcus spp. beta-hemolítico no rescaldo de pleuropneumonia em um cavalo. Relato de um caso / Pleural abscesses produced by Staphylococcus aureus and beta-haemolytic Streptococcus spp. as a sequel of pleuropneumonia in a horse. A case report

Los abscesos pleurales son una complicación frecuente en caballos con pleuroneumonía. Su tratamiento resulta difícil, requiere tiempo y dinero. Un caballo criollo argentino, castrado, de 13 años de edad presentó una pleuroneumonía grave que posteriormente se complicó con abscesos pleurales. El uso de anti-inflamatorios no esteroidales (fenilbutazona), una antibioterapia agresiva (ceftiofur, ampicilina y gentamicina), el drenaje eco-guiado y el lavado de los abscesos con una solución de yodopovidona al 0.05% permitió la recuperación exitosa del paciente de este reporte.

Pleural abscesses are a frequent complication in horses with pleuropneumonia. Their treatment is not easy, and requires time and it is expensive. A 13 years old creole argentine gelding presented a severe pleuropneumonia subsequently complicated with pleural abscess. The use of nonsteroidal antiinflammatory drugs (fenylbutazone), an aggressive antibiotherapy (ceftiofur, ampicillin, and gentamicin), echoguide drainage and lavage with yodopovidone to 0.05% of the abscesses permitted the successful recuperation of the patient of this case report.

Os abscessos pleurais são uma complicação comum em cavalos com pleuropneumonia. Seu tratamento é difícil e requer tempo e dinheiro. Um cavalo crioulo argentino, castrado, 13 anos de idade apresentou uma grave pleuropneumonia que posteriormente se tornou complicado com abscesso pleural. O uso de não-esteróides antiinflamatórios (fenilbutazona), uma agressiva terapia antibiótica (cetfiofur, ampicilina e gentamicina), eu drenagem eco-guiado e da lavagem dos abscessos com uma solução de yodopovidona de 0.05%, permitiu o sucesso da recuperação do paciente deste relato de caso.

Ocorrência de Actinobacillus pleuropneumoniae biótipo 1 em pulmões de suínos com pleuropneumonia no Norte de Portugal / Occurrence of Actinobacillus pleuropneumoniae biotype 1 in swine with pleuropneumonia in the North of Portugal

The isolation and identification of Actinobacillus pleuropneumoniae in swine lungs with pleuropneumonia in the North of Portugal were reported. A total of 127 swine lungs with and without lesions were examined. The system of lesions classification was based on a semi-quantitative method. Diagnosis was made by isolation and identification of the etiological agent in typical lesions. The occurrence of observed lesions was 75.6 percent and the occurrence of isolation of A. pleuropneumoniae was 19.7 percent. In 25 out of 96 (26.0 percent) lung samples with lesions of pleuropneumonia, A. pleuropneumoniae was isolated.

Otimização da técnica da PCR para a detecção de Actinobacillus pleuropneumoniae / Optimization of PCR technique for detection of Actinobacillus pleuropneumoniae

A utilização de métodos moleculares baseados em PCR é fundamental na detecção do Actinobacillus pleuropneumoniae, sendo capaz de identificar a infecção antes do estabelecimento da doença no rebanho. Estes métodos apresentam maior sensibilidade quando comparados com métodos tradicionais de isolamento bacteriano, mas podem sofrer influência de substâncias que reduzem a especificidade do teste e proporcionam o aparecimento de amplificações inespecíficas. No intuito de reduzir as amplificações inespecíficas, observadas quando aplicada a PCR para o gene cpx em amostras de tecido tonsilar, procedeu-se a otimização da técnica, na qual foram analisados o efeito do pré-cultivo bacteriano e as diferentes temperaturas de anelamento dos iniciadores e foi introduzido, no protocolo, um anticorpo que se liga na enzima Taq DNA Polimerase, aumentando a especificidade do teste. Paralelamente, foi realizado um experimento para verificar o efeito inibidor do tecido tonsilar sobre os resultados da PCR. Para isso, porções de tonsila de animais negativos para A. pleuropneumoniae foram contaminadas artificialmente com a amostra referência do sorotipo 5B. A adição do anticorpo para a enzima Taq DNA Polimerase e o aumento da temperatura de anelamento dos iniciadores para 57ºC diminuiu o aparecimento de amplificações inespecíficas. Os resultados obtidos no experimento demonstraram que o tecido tonsilar possui efeito inibidor nas amplificações da PCR. Além disso, a amplificação depende de, no mínimo, 675 UFC presentes na alíquota da amostra usada na PCR (equivalente a 1,35 x 10(5) UFC mL-1), assim, amostras de fragmentos de tecido de infecções iniciais e/ou com poucas células podem apresentar resultados falsos negativos.

The use of molecular methods based on PCR is important in Actinobacillus pleuropneumoniae detection, being able to identify the infection before the establishment of the disease in the herd. These methods have larger sensitivity when compared with traditional methods of bacteriological isolation, but they can suffer influence of substances that reduce the specificity of the test and resulting in inespecific amplifications. In order to reduce inespecific amplifications, observed when applied the PCR technique for the gene cpx in tonsil's tissue samples, the optimization was performed, in which different annealing temperatures were analyzed and introduced, in the technique, an antibody that binds to the enzyme Taq DNA Polimerase, increasing its specificity. In parallel, an experiment was performed in order to verify the inhibiting effect of the tonsil's tissue on the PCR results. For that, portions of tonsil from animals negative to the A. pleuropneumoniae were artificially contaminated with the reference sample of the sorotype 5B. The addition antibody for the enzyme Taq DNA Polimerase and the increase of the primers anneling temperature to 57ºC reduced the inespecific amplifications. The results obtained in the experiment demonstrated a possible inhibiting effect of the tonsil's tissue in the PCR amplifications. Besides, amplifications depend on at least 675 UFC present in the aliquot of samples that will be used in PCR (equivalent to 1.35 x 10(5) UFC mL-1), therefore, samples tissue's fragments in initial infections and/or with few cells can result in false-negative.

Reação em Cadeia da Polimerase (PCR) baseada no gene cpx para detecção de Actinobacillus pleuropneumoniae em suínos natural e experimentalmente infectados / Polymerase Chain Reaction (PCR) based on the cpx gene for detection of Actinobacillus pleuropneumoniae in natural and experimentally infected pigs

A pleuropneumonia suína é uma das mais importantes doenças respiratórias dos suínos, estando presente em todos os países produtores. Para o controle e o monitoramento da pleuropneumonia, é necessário o desenvolvimento de métodos rápidos e acurados de diagnóstico. Com o objetivo de validar a técnica da PCR, baseada no gene cpx de Actinobacillus pleuropneumoniae, em suínos sabidamente positivos, primeiramente foi realizada inoculação experimental com amostras de A. pleuropneumoniae sorotipo 5B e coletadas amostras por meio de suabe de tonsila, biópsia de tonsila e sangue para realização da técnica de PCR, isolamento bacteriológico e teste de ELISA, respectivamente. Posteriormente, estas técnicas foram aplicadas em suínos naturalmente infectados, em três rebanhos com diferentes situações sanitárias quanto à apresentação clínica da doença. De cada rebanho, foram analisados cinco grupos de suínos com idades diferentes, sendo coletado de cada animal biópsia de tonsila para isolamento bacteriológico e PCR e sangue para determinação do perfil sorológico. Os resultados obtidos na inoculação experimental confirmaram que, mesmo com o estabelecimento da infecção comprovada pelo isolamento bacteriológico, após o período de 45 dias, não foi possível detectar o agente pela técnica de PCR. Em animais naturalmente infectados, a técnica de PCR apresentou maior sensibilidade quando comparado com o isolamento. A associação entre PCR e ELISA demonstrou ser uma boa alternativa para definir a situação sanitária do rebanho quanto à infecção por A. pleuropneumoniae.

Swine pleuropneumonia is one of the most important pig respiratory diseases and has been found in all producer countries. For control and monitoring of pleuropneumonia, it is necessary the development of fast and specific methods of diagnosis. To validate PCR based on the cpx gene of Actinobacillus pleuropneumoniae in positive pigs, an experimental infection with A. pleuropneumoniae serotype 5B was performed and samples were obtained by tonsil swab, tonsil biopsy and blood for PCR, bacterial isolation and ELISA, respectively. These tests were then performed in naturally infected pigs from three herds with different sanitary situations of clinical disease. In each herd, five groups of different ages were analyzed. Tonsil biopsy for bacterial isolation and PCR and blood to determine the herd serological status was collected. The results obtained in the experimental infection confirmed that, even with the infection establishment, proved with bacterial isolation, it was not possible to detect the agent by PCR 45 days after infection. In naturally infected animals, PCR was more sensitive than bacterial isolation. The association between PCR and ELISA is a good alternative to define the herd sanitary status regarding the infection with A. pleuropneumoniae.

Padronização de três ELISAs polivalentes com lipopolissacarídeos de cadeia longa dos sorotipos 1 e 5, 2, 3 e 7 ou 10 e 12 de Actinobacillus pleuropneumoniae / Standardization of three polyvalent ELISA based on long chain lipopolysaccharides of serotypes 1 and 5, 2, 3 and 7, or 10 and 12 of Actinobacillus pleuropneumoniae

Três ELISAs polivalentes baseados em lipopolissacarídeos de cadeia longa (LPS-CL) foram estabelecidos para detectar anticorpos para todos os sorotipos prevalentes de Actinobacillus pleuropneumoniae. Foram testadas amostras provenientes do banco de soros de suínos experimentalmente inoculados com todos os sorotipos de A. pleuropneumoniae. Os ELISAs foram sensíveis à detecção de anticorpos contra todos os LPS-CL. Foram observadas reações cruzadas no ELISA polivalente produzido com os sorotipos 1 e 5, com anti-soros específicos para os sorotipos 9 e 11, pois os sorotipos 1, 9 e 11 apresentaram antígenos somáticos comuns. No polivalente com os sorotipos 2, 3 e 7, observaram-se reações com anti-soros dos sorotipos 4, 6 e 8, devido à presença de antígenos somáticos entre os sorotipos 3, 6 e 8 e entre os sorotipos 4 e 7. Amostras de soros de animais infectados com Mycoplasma hyopneumoniae, Mycoplasma flocculare e Haemophilus parasuis, agentes que acometem o sistema respiratório dos suínos, não apresentaram reações cruzadas com os antígenos baseados em LPS-CL.

Three polyvalent ELISA based on long chain lipopolysaccharides (LC-LPS) were established to detect all prevalent serotypes of Actinobacillus pleuropneumoniae. Samples from a serum bank of experimentally inoculated animals with all serotypes of A. pleuropneumoniae were tested. Antibodies specific to LC-LPS of each serotype were detected. Cross-reactions were observed in the polyvalent ELISA produced with serotypes 1 and 5, with specific antisera to serotypes 9 and 11 due to common somatic antigens presence in serotypes 1, 9, and 11. In the polyvalent with serotypes 2, 3 and 7 reactions were observed with antisera of serotypes 4, 6, and 8, due to the presence of somatic antigens in serotypes 3, 6, and 8 and serotypes 4 and 7. Experimentally infected animals with respiratory agents of swine Mycoplasma hyopneumoniae, Mycoplasma flocculare, and Haemophilus parasuis did not present cross-reactions with the antigens based on LC-LPS.

Pleuroneumonías pediátricas en Hospital Clínico Regional Valdivia (HCRV): manejo médico-quirúrgico / Clinico-surgical management in children with pleuropneumonia: Hospital Clinico Regional Valdivia experience

Se define pleuroneumonía a las neumonías que presentan compromiso inflamatorio pleural, asociado o no a derrame pleural. Su tratamiento se basa en el empleo juicioso y precoz de antibióticos y/o drenaje pleural, y/o fibrinolíticos; y/o debridación por videotoracoscopía; y/o decorticación por toracotomÍa. El objetivo de nuestro trabajo fue determinar el comportamiento de las pleuroneumonías en HCRV, en pacientes menores de 15 años, caracterizando la clínica, su evolución, tratamientos efectuados y la efectividad de éstos; mediante un estudio descriptivo, retrospectivo de las fichas clínicas con diagnóstico de egreso de Pleuroneumonía en el HCRV, entre enero del 2000 y diciembre del 2005. Se registraron 75 casos de Pleuroneumonía en un número de 6 a 16 casos por año; en edades entre 6 meses y 14 años 10 meses; con predominio del grupo etario Lactantes. Uno de cada cuatro pacientes requirió algún tipo de intervención quirúrgica. Se concluye que las pleuroneumonías si bien son infrecuentes, significan una alta morbilidad para los pacientes y su manejo requiere de protocolos que permitan un actuar oportuno y seguro.

A hematogenic pleuropneumonia caused by postoperative septic thrombophlebitis in a Thoroughbred gelding

A 7-year-old Thoroughbred gelding was admitted to Equine Hospital, Korea Racing Association for evaluation and treatment of colic. Based on the size and duration of the large colonic and cecal impaction, a routine ventral midline celiotomy and large colon enterotomy were performed to relieve the impaction. Six days following surgery the gelding exhibited signs of lethargy, fever, inappetence and diarrhea. Eleven days following surgery, the jugular veins showed a marked thrombophlebitis. On the sixteenth day of hospitalization the gelding died suddenly. Upon physical examination, the horse was febrile, tachycardic and tachypnoeic. Thoracic excursion appeared to be increased; however, no abnormal lung sounds were detected. No cough or nasal discharge was present. Hematology revealed neutrophilic leukocytosis. Serum biochemistry was normal but plasma fibrinogen increased. In necropsy, fibrinopurulent fluid was present in the thoracic cavity. There were firm adhesions between visceral pleura and thoracic wall. White, mixed and red thrombi were formed in both jugular veins from the insertion point of IV catheter. Histopathological examination showed fibrinopurulent inflammation and vascular thrombosis in the lung. The pleura showed edematous thickening and severe congestion. The clinicopathological and pathological findings suggest that septic thrombi associated with septic thrombophlebitis metastasized into the pulmonary circulation and were entrapped in the pulmonary parenchyma and provoked pleuropneumonia.

Presencia de fístula broncopleural en niños con patología pulmonar: descripción de 9 casos clínicos / Bronchopulmonary fistula in children: a description of 9 clinical cases

Introducción: La fístula brocopleural (FBP) es una comunicación anormal que se establece entre el árbol bronquial y el espacio pleural, que se presenta en la evolución de enfermedades respiratorias graves, como complicación de intervenciones quirúrgicasdel pulmón o de procedimientos con riesgos de trauma pulmonar. Objetivo: Presentar las características clínicasde un grupo de pacientes con FBP. Pacientes y Método: Se presenta una serie clínica de 9 pacientes con FBP (7 mujeres y 2 hombres) con una edad promedio de 2 años 1 mes (rango entre 7 días y 7 años). Resultados: En 6 pacientes, la FBP apareció en la evolución de una enfermedad respiratoria grave (pleuroneumoníacomplicada en 4, obstrucción bronquial severa en 1 y síndrome de distrés respiratorio agudo en 1). En los otros 3 pacientes, la FBP fue complicación de una lobectomía indicada por malformaciones pulmonares congénitas y bronquiectasias. El cuadro clínico se caracterizó por el deterioro respiratorio debido al desarrollo de un neumotórax a tensión (4 casos) o neumotórax recurrente (5 pacientes). En 5 casos la FBP se presentó con el paciente en ventilación mecánica, y en 2 pacientes con pleroneumonía, la FBP se asoció al trauma pulmonar producido por la instalación del tubo de drenaje. El tratamiento de la FBP consistió en drenaje pleural sin aspiración y en los pacientes en ventilación mecánica se ajustaron las presiones para lograr la menor fuga posible a través de la FBP. En 2 casos se requirió ventilación de alta frecuencia. En 4 pacientes con enfermedad pulmonar grave y en 2 casos de lobectomía, la FBP se resolvió con tratamiento quirúrgico. No hubo fallecidos en esta serie. Conclusión: En nuestra experiencia clínica la FBP en pacientes pediátricos se presenta durante la evolución de una enfermedad pulmonar grave en ventilación mecánica, o como complicación de una cirugía pulmonar

Síndrome hemolítico urémico y streptococcus pneumoniae / Hemolytic uremic syndrome after an infection caused by streptococcus pneumoniae

Hemolytic-uremic syndrome (HUS) is an uncommon complication of pneumococcal infection. Highly suggesting findings in a patient with Streptococcus pneumoniae infection are: microangyopatic hemolytic anemia, thrombocytopenia and acute renal failure. We report a 41 years old woman, admitted to the hospital due to a severe pneumonia, that required the surgical drainage of an empyema. On admission, a drop in packed red cell volume from 41 to 25 percent, the presence of schistocytes in the blood smear, an elevation of LDH to 1,700 IU/L, a fall in haptoglobin to 5.8 mg/dL and a thrombocytopenia of 72,000 per mm3 were detected. These alterations coincided with an oliguric acute renal failure. She was treated with hemodialysis and the hemolytic syndrome was managed with plasmapheresis. She was discharged 35 days after admission and in the follow up, after 2.5 months, her serum creatinine is 1.2 mg/dL and her packed red cell volume is 41 percent