Contagious Bovine Pleuropneumonia: A Comprehensive Overview.

Contagious bovine pleuropneumonia (CBPP) is a respiratory disease of cattle that is listed as notifiable by the World Organization for Animal Health. It is endemic in sub-Saharan Africa and causes important productivity losses due to the high mortality and morbidity rates. CBPP is caused by Mycoplasma mycoides subsp. mycoides (Mmm) and is characterized by severe fibrinous bronchopneumonia and pleural effusion during the acute to subacute stages and by pulmonary sequestra in chronic cases. Additional lesions can be detected in the kidneys and in the carpal and tarsal joints of calves. Mmm infection occurs through the inhalation of infected aerosol droplets. After the colonization of bronchioles and alveoli, Mmm invades blood and lymphatic vessels and causes vasculitis. Moreover, Mmm can be occasionally demonstrated in blood and in a variety of other tissues. In the lung, Mmm antigen is commonly detected on bronchiolar and alveolar epithelial cells, in lung phagocytic cells, within the wall of blood and lymphatic vessels, inside necrotic areas, and within tertiary lymphoid follicles. Mmm antigen can also be present in the cytoplasm of macrophages within lymph node sinuses, in the germinal center of lymphoid follicles, in glomerular endothelial cells, and in renal tubules. A complete pathological examination is of great value for a rapid presumptive diagnosis, but laboratory investigations are mandatory for definitive diagnosis. The purpose of this review is to describe the main features of CBPP including the causative agent, history, geographic distribution, epidemiology, clinical course, diagnosis, and control. A special focus is placed on gross and microscopic lesions in order to familiarize veterinarians with the pathology and pathogenesis of CBPP.

Disseminated Mycobacterium kansasii infection in a white-tailed deer and implications for public and livestock health.

We document a case of Mycobacterium kansasii, a rare, zoonotic bacterium, in a white-tailed deer (WTD; Odocoileus virginianus) in East Baton Rouge Parish, Louisiana. Grossly, the deer had fibrinous pleuropneumonia with yellow, mineralized nodules scattered throughout the lungs and extending to the pleura. The kidneys were enlarged and had numerous pale foci in the cortex. Microscopically, the pulmonary architecture was replaced by variably sized, multifocal-to-coalescing granulomas with peripheral histiocytes and fewer multinucleate giant cells, and necrotic centers with mineralization and hemorrhage. The latter rarely contained one to a few acid-fast, slender, 7-µm long bacteria, for which beaded morphology was sometimes evident. Similar acid-fast bacteria were also within histiocytes in the kidney. PCR assay of fresh lung sample and subsequent sequencing revealed a non-tuberculosis mycobacterium, M. kansasii. These lesions were similar to those that result from infection with M. bovis in WTD. Both M. bovis and M. kansasii are zoonotic. WTD are a reservoir of M. bovis, which is a major concern in regions in which WTD and cattle can come into close contact.

Comparative evaluation of different therapeutic protocols for contagious caprine pleuropneumonia in Himalayan Pashmina goats.

Therapeutic management of contagious caprine pleuroneumonia (CCPP) involves mostly the use of oxytetracycline followed by enrofloxacin and rarely tylosin. In many parts of the world including India, the former antibiotics are commonly available than the latter. Therefore, prolonged use of the same leads to the development of antibiotic resistance and decreased efficacy of drug. Besides, inflammatory and allergic pathogenesis of CCPP envisages combination therapy. In this study, we evaluated the effectiveness of the combination therapy using different antibiotics (oxytetracycyline @ 10: group I, enrofloxacin @ 5 group II, and tylosin: group III, @ 10 mg/kg body weight), along with anti-inflammatory (meloxicam @ 0.5 mg/kg) and anti-allergic (pheneramine maleate @ 1.0 mg/kg) drugs. These drugs were given intramuscularly at the interval of 48 h for four times in three test groups (n = 10) of Pashmina goats, viz. groups I, II, and III, respectively, affected with CCPP. Group IV (n = 10) was kept as healthy control when group V (n = 10) treated with oxytetracycline @ 10 mg/kg alone was used as positive control. Clinical signs, clinical parameters, pro-inflammatory cytokine (tumor necrosis factor alpha (TNF-α)), and oxidative stress indices (total oxidant status (TOS), total antioxidant status (TAS)) were evaluated at hours 0, 48, 96, and 144 of experimental trial. Tylosin-based combination therapy resulted in a rapid and favorable recovery resulting in restoration of normal body temperature (102.46 ± 0.31 °F), respiration rate (16.30 ± 0.79 per minute), and heart rate (89.50 ± 2.63 per minute) compared to the oxytetracycline (102.95 ± 0.13, 21.30 ± 1.12, 86.00 ± 2.33, respectively) and enrofloxacin (102.97 ± 0.19, 21.00 ± 1.25, 90.00 ± 2.58, respectively) treated groups. By hour 144, all the groups showed restoration of clinical parameters of normal health and diminishing signs of CCPP, viz. fever, dyspnea, coughing, nasal discharge, weakness, and pleurodynia. Significant (P ≤ 0.05) decrease in levels of TNF-α and non-significant (P > 0.05) decrease in levels of TOS and an increase in levels of TAS were noted from hour 0 to 144 in all the test groups. Within the groups, no significant (P > 0.05) change was noted in TNF-α, TOS, and TAS levels; however, TNF-α levels were comparatively lower in group III. Hematological parameters did not differ significantly (P > 0.05). From these findings, it can be inferred that tylosin-based combination therapy is relatively better for early, rapid, and safe recovery besides minimizing inflammatory and oxidative cascade in CCPP affected Pashmina goats compared to oxytetracycline- and enrofloxacin-based therapies.

Activation of Porcine Alveolar Macrophages by Actinobacillus pleuropneumoniae Lipopolysaccharide via the Toll-Like Receptor 4/NF-κB-Mediated Pathway.

Actinobacillus pleuropneumoniae is the causative agent of porcine contagious pleuropneumonia. Overproduction of proinflammatory cytokines, like interleukin-1ß (IL-1ß), IL-6, tumor necrosis factor alpha, and resistin, in the lung is an important feature of A. pleuropneumoniae infection. These proinflammatory cytokines enhance inflammatory and immunological responses. However, the mechanism that leads to cytokine production remains unclear. As a major virulence factor of A. pleuropneumoniae, lipopolysaccharide (LPS) may act as a potent stimulator of Toll-like receptor 4 (TLR4), triggering a number of intracellular signaling pathways that lead to the synthesis of proinflammatory cytokines. Porcine alveolar macrophages (PAMs) are the first line of defense against pathogenic microbes during pathogen invasion. The results of the present study demonstrate that A. pleuropneumoniae LPS induces PAMs to produce inflammatory cytokines in time- and dose-dependent manners. Moreover, PAMs were activated by A. pleuropneumoniae LPS, resulting in upregulation of signaling molecules, including TLR4, MyD88, TRIF-related adaptor molecule, and NF-κB. In contrast, the activation effects of A. pleuropneumoniae LPS on PAMs could be suppressed by specific inhibitors, like small interfering RNA and Bay11-7082. Taken together, our data indicate that A. pleuropneumoniae LPS can induce PAMs to produce proinflammatory cytokines via the TLR4/NF-κB-mediated pathway. These findings partially reveal the mechanism of the overproduction of proinflammatory cytokines in the lungs of swine with A. pleuropneumoniae infection and may provide targets for the prevention of A. pleuropneumoniae-induced pneumonia. All the data could be used as a reference for the pathogenesis of respiratory infection.

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...

Nonsurgical resolution of caudal mediastinal paraesophageal abscess in a cat.

A one-year-old, castrated male domestic short hair cat was admitted with a history of anorexia, regurgitation and pyrexia for two days. Fever and leukocytosis were identified. There were a large soft tissue density oval mass in the caudal mediastinum on thoracic radiographs, a fluid-filled oval mass in the caudal mediastinum on ultrasonography, and left-sided and ventrally displaced and compressed esophagus on esophagram. On esophageal endoscopy, there were no esophageal abnormalities. CT findings with a fluid filled mass with rim enhancement indicated a caudal mediastinal paraesophageal abscess. The patient was treated with oral antibiotics, because the owner declined percutaneous drainage and surgery. The patient was admitted on emergency with severe respiratory distress; and ruptured abscess and deteriorated pleuropneumonia were suspected. With intensive hospitalization care and additional antibiotic therapy, the patient had full recovery.

Isolation and phylogenetic characterization of Streptococcus halichoeri from a European badger (Meles meles) with pyogranulomatous pleuropneumonia.

Clinical and pathological studies in European badgers (Meles meles) are limited. Badgers play a significant role in the epidemiology of bovine tuberculosis (TB) in some countries and an accurate diagnosis is needed for this infection. However, the lesions of bovine TB are similar to those associated with other pathogens, making pathological diagnosis difficult. In the present study, Streptococcus halichoeri was isolated from a European badger with pyogranulomatous pleuropneumonia and suspected of having tuberculosis. TB and other pathogens able to induce similar lesions were ruled out. Comparative 16S rRNA and rpoB gene sequencing studies showed an identity of 99.51% and 98.28%, respectively, with S. halichoeri. This report represents the third description of this bacterium and the first in an animal species other than the grey seal (Halichoerus grypus). It also shows that S. halichoeri can be associated with a pathological process characterized by granulomatous inflammation and resembling tuberculosis.

Infection dynamics and acute phase response of an Actinobacillus pleuropneumoniae field isolate of moderate virulence in pigs.

Actinobacillus pleuropneumoniae, the causative agent of porcine contagious pleuropneumonia (PCP), causes significant economic losses associated mainly with growth stunting of animals. Although serotypes can be distinguished according to their virulence, most of the studies are focused in A. pleuropneumoniae infections with virulent serotypes. There is little information regarding the role of acute phase proteins (APPs) and proinflammatory cytokines in infections with isolates of mild or moderate virulence. Thus, the present study aims to evaluate the kinetics of infection with an A. pleuropneumoniae serotype 6 (Ap6) field isolate of moderate virulence and the changes in the serum concentration of specific antibodies and different APPs and proinflammatory cytokines. Control animals showed no clinical signs or lesions throughout the study. Infected animals showed increased rectal temperature, respiratory distress and depression from 24hpi, and typical gross and microscopic lesions of PCP from 6hpi onwards. Ap6 was isolated from nasal swabs of four out of five inoculated animals at 24hpi, and from nasal swabs, tonsil and lung samples from all inoculated animals at 72hpi. Specific antibodies against Ap6 or changes in the serum concentration of IL-1ß, IL-10 and TNF-α were not detected throughout the study. The serum concentration of IL-6 increased from 6hpi as well as serum A amyloid, C-reactive protein and haptoglobin from 24hpi onwards. Our results highlight the onset of the acute phase response after the infection with a field isolate of A. pleuropneumoniae of moderate virulence from 24hpi onwards which may be of interest in the study of the pathogenesis of this disease.

Blood values of captive beira antelope (Dorcatragus megalotis) prior to and during an outbreak of fibrinous pleuropneumonia syndrome (FPPS).

Currently the only captive population of beira antelope (Dorcatragus megalotis) is held at the Al Wabra Wildlife Preservation, Qatar. An outbreak of a severe respiratory disease--fibrinous pleuropneumonia syndrome, most likely caused by Mycoplasma ovipneumoniae--led to a marked population decline. Reactive systemic inflammatory (AA) amyloidosis was noted as a chronic manifestation of the disease. Blood samples had been collected for biochemistry and hematology baseline values prior to the outbreak. Population-level changes were analyzed before and during the course of the outbreak in selected blood parameters (white blood cells [WBC], blood urea nitrogen [BUN], and creatinine). The annual population WBC increased and decreased concurrently with the population size, with a significant correlation between the two measures (R = 0.92; P = 0.001). Both BUN and creatinine values were higher during the outbreak. These values peaked at the same time as mortality, which was 1 yr after the WBC peak. These changes were interpreted as the transition from an acute disease with a primary respiratory manifestation into a chronic condition where renal amyloidosis led to chronic renal failure and death. Also, elevated liver values in diseased animals were attributed to amyloidosis. Parallels to a literature report on a lung disease complex caused by M. ovipneumoniae in bighorn sheep (Ovis canadensis) were found. Trends in population-level blood values of the beira antelopes implicate amyloidosis as a significant, long-term consequence of the putative Mycoplasma infection.

The RNA chaperone Hfq promotes fitness of Actinobacillus pleuropneumoniae during porcine pleuropneumonia.

Actinobacillus pleuropneumoniae is the etiological agent of porcine pleuropneumonia, an economically important disease of pigs. The hfq gene in A. pleuropneumoniae, encoding the RNA chaperone and posttranscriptional regulator Hfq, is upregulated during infection of porcine lungs. To investigate the role of this in vivo-induced gene in A. pleuropneumoniae, an hfq mutant strain was constructed. The hfq mutant was defective in biofilm formation on abiotic surfaces. The level of pgaC transcript, encoding the biosynthesis of poly-ß-1,6-N-acetylglucosamine (PNAG), a major biofilm matrix component, was lower and PNAG content was 10-fold lower in the hfq mutant than in the wild-type strain. When outer membrane proteins were examined, cysteine synthase, implicated in resistance to oxidative stress and tellurite, was not found at detectable levels in the absence of Hfq. The hfq mutant displayed enhanced sensitivity to superoxide generated by methyl viologen and tellurite. These phenotypes were readily reversed by complementation with the hfq gene expressed from its native promoter. The role of Hfq in the fitness of A. pleuropneumoniae was assessed in a natural host infection model. The hfq mutant failed to colonize porcine lungs and was outcompeted by the wild-type strain (median competitive index of 2 × 10(-5)). Our data demonstrate that the in vivo-induced gene hfq is involved in the regulation of PNAG-dependent biofilm formation, resistance to superoxide stress, and the fitness and virulence of A. pleuropneumoniae in pigs and begin to elucidate the role of an in vivo-induced gene in the pathogenesis of pleuropneumonia.

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.

Standing lateral thoracotomy in horses: indications, complications, and outcomes.

OBJECTIVE: To describe the indications for, complications arising from, and outcome of horses that had standing lateral thoracotomy for pleural or pericardial disease. STUDY DESIGN: Case series. ANIMALS: Horses (n=16). METHODS: Medical records (January 1990-December 2008) of sedated standing horses that had lateral thoracotomy were reviewed. Clinical and surgical findings, perioperative and short-term complications were recorded. Long-term (>6 months) outcome was determined through telephone conversations with owners and veterinarians. RESULTS: Mean (±SD) horse age was 6.6±5.3 years (range, 1-15 years). Thoracotomy was most commonly for treatment of recurrent or chronic pleural infection (94%). Anaerobic bacteria were frequently isolated from pleural effusion associated with pleuropneumonia (63%). Right lateral intercostal thoracotomy was performed in 13 horses (82%); intercostal muscle myectomy in 5 horses; and rib resection in 1. Perioperative complications were hemipneumothorax (2 horses) and short-term complications included cellulitis and abscessation of the thoracotomy site (6 horses). Fourteen (88%) horses survived to discharge and 46% of horses that survived returned to their previous level of athletic activity. CONCLUSIONS: Lateral thoracotomy is well tolerated by standing sedated horses with minimal perioperative and short-term complications. CLINICAL RELEVANCE: Standing lateral thoracotomy should be considered for the treatment of complicated pleuropneumonia in horses.

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.

Prevalence of granulomatous pleuropneumonia associated with Actinobacillus pleuropneumoniae serotype 2 in slaughter pigs.

A total of 14,818 slaughtered pigs were examined macroscopically. Of these, 25 pigs with porcine pleuropneumonia were collected and the relations among Actinobacillus spp. and granulomatous lesions in organs (lungs and tonsils) were evaluated. In the lungs, only Actinobacillus pleuropneumoniae serotype 2 was isolated from 20 of the pigs. Histologically, granulomatous pneumonia with A. pleuropneumoniae antigen was detected in 8 of the pigs. The antigen was visible in the centers of the lesions along with asteroid bodies, epithelioid cells and multinucleated giant cells. In the tonsils, granulomatous lesions were not detected, although A. pleuropneumoniae serotype 2 (5 pigs), serotype 7 (1 pig), Actinobacillus porcitonsillarum (1 pig) and Actinobacillus minor (1 pig) were isolated. The present survey suggests that multifocal granulomatous pneumonia in slaughter pigs could be highly associated with A. pleuropneumoniae serotype 2 infection.

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.

Multifocal granulomatous hepatitis caused by Actinobacillus pleuropneumoniae serotype 2 in slaughter pigs.

In a survey of 66 894 slaughter pigs, 11 animals from three farms were found to have multifocal granulomatous lesions in the liver, caused by Actinobacillus pleuropneumoniae serotype 2. The lesions consisted of epithelioid cells and multinucleated giant cells, with asteroid bodies and discernible gram-negative bacteria. Lymph nodes and spleen were occasionally affected. The results suggested that haematogenous spread had occurred from pre-existing pulmonary infections.