Synergic Effect of Brachyspira hyodysenteriae and Lawsonia intracellularis Coinfection: Anatomopathological and Microbiome Evaluation.

Brachyspira hyodysenteriae and Lawsonia intracellularis coinfection has been observed in the diagnostic routine; however, no studies have evaluated their interaction. This study aimed to characterize lesions and possible synergisms in experimentally infected pigs. Four groups of piglets, coinfection (CO), B. hyodysenteriae (BRA), L. intracellularis (LAW), and negative control (NEG), were used. Clinical signals were evaluated, and fecal samples were collected for qPCR. At 21 days post infection (dpi), all animals were euthanized. Gross lesions, bacterial isolation, histopathology, immunohistochemistry, and fecal microbiome analyses were performed. Diarrhea started at 12 dpi, affecting 11/12 pigs in the CO group and 5/11 pigs in the BRA group. Histopathological lesions were significantly more severe in the CO than the other groups. B. hyodysenteriae was isolated from 11/12 pigs in CO and 5/11 BRA groups. Pigs started shedding L. intracellularis at 3 dpi, and all inoculated pigs tested positive on day 21. A total of 10/12 CO and 7/11 BRA animals tested positive for B. hyodysenteriae by qPCR. A relatively low abundance of microbiota was observed in the CO group. Clinical signs and macroscopic and microscopic lesions were significantly more severe in the CO group compared to the other groups. The presence of L. intracellularis in the CO group increased the severity of swine dysentery.

Experimental Infection of Pigs with a ST 245 Brachyspira hyodysenteriae Isolated from an Asymptomatic Pig in a Herd with No History of Swine Dysentery.

Swine dysentery (SD) is characterized by a severe mucohemorrhagic colitis caused by infection with Brachyspira species. In infected herds the disease causes considerable financial loss due to mortality, slow growth rates, poor feed conversion, and costs of treatment. B. hyodysenteriae is the most common etiological agent of SD and infection is usually associated with disease. However, isolated reports have described low pathogenic strains of B. hyodysenteriae. The aim of this study was to describe an experimental infection trial using a subclinical B. hyodysenteriae isolated from an animal without clinical signs and from a disease-free herd, to evaluate the pathogenicity and clinical pathological characteristics compared to a highly clinical isolate. Forty-eight 5-week-old pigs were divided into three groups: control, clinical and the subclinical isolates. The first detection/isolation of B. hyodysenteriae in samples of the animals challenged with a known clinical B. hyodysenteriae strain (clinical group) occurred 5th day post inoculation. Considering the whole period of the study, 11/16 animals from this group were qPCR positive in fecal samples, and diarrhea was observed in 10/16 pigs. In the subclinical isolate group, one animal had diarrhea. There were SD large intestine lesions in 3 animals at necropsy and positive B. hyodysenteriae isolation in 7/15 samples of the subclinical group. In the control group, no diarrhea, gross/microscopic lesions, or qPCR positivity were observed. Clinical signs, bacterial isolation, macroscopic and histologic lesions were significantly difference among groups, demonstrating low pathogenicity of the subclinical isolate in susceptible pigs.

Use of oral fluids to detect anti Lawsonia intracellularis antibodies in experimentally infected pigs / Utilização de fluidos orais para detecção de anticorpos anti Lawsonia intracellularis em suínos experimentalmente infectados

Several pathogens and antibodies derived from serum or produced in tissues associated with the oral cavity are present in the oral fluid (OF). Considering the applicability of this alternative sample, recent studies in veterinary medicine have tested OF as a replacement for serum in diagnostic assays. The aim of this study was to standardize the immunoperoxidase monolayer assay (IPMA) to detect anti-Lawsonia intracellularis immunoglobulin A (IgA) and immunoglobulin G (IgG) in OF samples from experimentally infected pigs. Sixty-two pigs were divided into two groups: control (T1, n=30) and inoculated with L. intracellularis (T2, n=32). Blood, OF and fecal samples were collected at 0, 7, 14, 21, 28 and 42 days post-inoculation (dpi). Some adaptations of the standard technique for serum were made to IPMA for the detection of IgA and IgG in OF. The IPMA showed high specificity and sensitivity for serum samples and high specificity and moderate sensitivity for the detection of IgA and IgG in OF. There was high agreement between the results of serum IgG and OF IgA and IgG. Based on our results, oral fluid samples may be used for the evaluation and determination of anti-L. intracellularis antibodies in pigs, but not for individual diagnosis of swine proliferative enteropathy.(AU)

Vários patógenos e anticorpos derivados do soro ou produzidos em tecidos associados a cavidade oral estão presentes no fluido oral (FO). Considerando a aplicabilidade dessa amostra alternativa, estudos recentes em medicina veterinária têm testado o FO como substituto do soro para testes diagnósticos. O objetivo desse estudo foi padronizar a imunoperoxidase em monocamada de célula (IPMC) para a detecção de imunoglobulina A e imunoglobulina G anti-Lawsonia intracellularis em amostras de FO de suínos experimentalmente infectados. Um total de 62 suínos foram divididos em dois grupos: controle (T1, n=30) e inoculados com L. intracellularis (T2, n=32). Sangue, FO e amostras de fezes foram coletados aos 0, 7,14, 21, 28 e 42 dias após a inoculação (dpi). Algumas adaptações da técnica foram realizadas na técnica padrão da IPMC para a detecção de IgA e IgG. A IPMC demostrou alta especificidade e sensibilidade para amostras de soro e alta especificidade de moderada sensibilidade para a detecção de IgA e IgG em FO. Houve alta concordância entre resultados de detecção de IgG em soro com a IgA e IgG em amostras de FO. Baseado em nossos resultados, amostras de fluido oral podem ser usadas em avaliações e detecção de anticorpos anti-L. intracellularis em suínos, porém não de forma individual.(AU)

Oral fluid for detection of exposure to Lawsonia intracellularis in naturally infected pigs.

To demonstrate the utility of oral fluid (OF) for indirect diagnostic detection of Lawsonia intracellularis (Li), 15 pig farms were studied. Serum and fecal samples were collected from 20 animals from five different age groups on each farm. OF samples were collected from animals in two pens of the same age groups. Serum and OF samples were analyzed in an immunoperoxidase in monolayer assay (IPMA) for the detection of anti-Li immunoglobulin G (IgG) and A (IgA). Compatible results were found between PCR and IgG in OF in four of the five ages evaluated. Simultaneous detection of IgG in serum and OF was mainly observed on farms showing clinical signs suggestive of porcine proliferative enteropathy (PPE). These findings demonstrate the potential usefulness of OF in detecting anti-Li antibodies as a diagnostic tool that can be used to monitor PPE in herds with clinical signs compatible with the disease.

Mycoplasma hyorhinis infection in early cases of mycoplasmal pneumonia in swine and evaluation of diagnostic assays / Infecção por Mycoplasma hyorhinis em casos precoces de pneumonia micoplásmica em suínos e comparação entre técnicas diagnósticas

Mycoplasmal pneumonia is an important disease in the pig industry. Due to the controversial role of Mycoplasma hyorhinis in this disease, confirmation of the presence of this bacterium and the identification of its roles in respiratory disease remain major challenges. The objectives of this study were to evaluate the presence of M. hyorhinis in early cases of mycoplasmal pneumonia and to determine the usefulness of fluorescent in situ hybridization (FISH) for the diagnosis of respiratory mycoplasmosis in naturally infected pigs. Ninety M. hyopneumoniae and/or M. hyorhinis-infected lung tissue samples based on diagnostic mosaic (DM) were used. The average age of the animals was 116 and 57 days (P<0.01) for groups 1 (positive-M. hyopneumoniae only) and 2 (positive-M. hyorhinis only), respectively. These findings suggest that development of lesions caused by M. hyorhinis occurs earlier than for M. hyopneumoniae. Using the DM as the gold standard, the sensitivity and specificity of FISH for M. hyopneumoniae were 75 and 100%, respectively, and were 40 and 73.3% for the immunohistochemistry (IHC). The sensitivity and specificity of FISH for M. hyorhinis were 76.7 and 100%, respectively. These findings demonstrate that FISH can be a useful tool for diagnosing mycoplasmosis. Viral antigens (PCV2 or influenza A) were detected in 53.3% (16/30) of the samples in group 2 (M. hyorhinis-PCR positive) and 13.3% (4/30) of the samples in group 1 (M. hyopneumoniae-PCR positive). This finding indicates that the association of M. hyorhinis and viral infection in nursery pigs likely starts due to a viral immunosuppressive condition.(AU)

A pneumonia micoplásmica causada por bactérias do gênero Mycoplasma é uma enfermidade de grande importância para indústria suinícola, sendo ainda controverso o papel desempenhado por Mycoplasma hyorhinis nessa doença. A confirmação da presença dessas bactérias bem como a identificação de seus papéis em doenças respiratórias continua sendo um grande desafio. Os objetivos desse estudo foram comparar diferentes técnicas, em especial a de hibridização fluorescente in situ (FISH), para diagnóstico de micoplasmoses respiratória em suínos naturalmente infectados e avaliar a presença do M. hyorhinis em casos precoces de pneumonia micoplásmica. Foram utilizadas 90 amostras de tecido pulmonar infectado para cada um ou ambos os agentes (M. hyopneumoniae e M. hyorhinis) determinados pelo mosaico de diagnóstico (sinais clínicos, lesões macroscópicas e microscópicas e pela PCR). No grupo de animais positivos pela PCR apenas para M. hyorhinis (Grupo 2) a média da idade foi de 57,32 dias e no grupo apenas positivo para M. hyopneumoniae (Grupo 1) a média foi de 116,31 dias (P<0,01). Estes achados sugerem que a colonização e o aparecimento de lesões causadas pelo M. hyorhinis seja mais precoce do que aquelas causadas pelo M. hyopneumoniae. As alterações microscópicas foram estatisticamente (P<0,01) mais intensas no grupo 1 do que no grupo 2. Usando o mosaico de diagnóstico como padrão ouro, a sensibilidade e especificidade na FISH para M. hyopneumoniae foi de 75 e 100%, respectivamente, e 40 e 73,3%, na imuno-histoquímica. A sensibilidade e especificidade da FISH para M. hyorhinis foi de 76,7 e 100%. Esses valores demonstram que a FISH pode ser uma ferramenta útil para diagnóstico de micoplasmoses. Foi detectada a presença de agentes virais (PCV2 ou influenza) em 53,3% das amostras do grupo 2 (M. hyorhinis) e em 13,3% das amostras do grupo 1 (M. hyopneumoniae).(AU)

Minimum inhibitory concentration of Brazilian Brachyspira hyodysenteriae strains / Concentração inibitória mínima de cepas Brachyspira hyodysenteriae brasileiras

The objectives of this study were to characterize Brachyspira hyodysenteriae isolates and to evaluate the antimicrobial susceptibility patterns of strains obtained from pigs in Brazil based on the minimal inhibitory concentration test (MIC). The MIC was performed for 22 B. hyodysenteriae isolates obtained from 2011 to 2013 using the following antimicrobial drugs: tylosin, tiamulin, valnemulin, doxycycline, lincomycin and tylvalosin. Outbreaks of swine dysentery were diagnosed based on clinical presentation, bacterial isolation, gross and microscopic lesions, duplex PCR for B. hyodysenteriae and B. pilosicoli and nox gene sequencing. All obtained MIC values were consistently higher or equal to the microbiological cut-off described in the literature. The MIC 90 values for the tested drugs were 8µg/ml for doxycycline, >4µg/ml for valnemulin, 8µg/ml for tiamulin, 32µg/ml for tylvalosin, >64µg/ml for lincomycin and >128µg/ml for tylosin. These results largely corroborate those reported in the literature. Tiamulin, doxycycline and tylvalosin showed the lowest MIC results. All of the samples subjected to phylogenetic analysis based on the nox gene sequence exhibited similar results, showing 100% identity to B. hyodysenteriae. This is the first study describing the MIC pattern of B. hyodysenteriae isolated in Brazil.(AU)

Os objetivos deste trabalho foram a caracterização de isolados de Brachyspira hyodysenteriae e avaliar os padrões de sensibilidade antimicrobiana de isolados obtidos a partir de suínos no Brasil com base no teste de concentração inibitória mínima (MIC). A MIC foi realizada em 22 isolados de B. hyodysenteriae obtidos entre 2011 a 2013 usando os seguintes antimicrobianos: tilosina, tiamulina, valnemulina, doxiciclina, lincomicina e tilvalosina. Surtos de disenteria suína foram diagnosticados com base na apresentação clínica, isolamento bacteriano, lesões macroscópicas e microscópicas, PCR duplex para B. hyodysenteriae e B. pilosicoli e sequenciamento do gene nox. Todos os valores de MIC obtidos foram consistentemente mais elevados ou igual ao ponto de corte microbiológica descrito na literatura. Os valores de MIC 90 para os fármacos testados foram de 8 µg / mL para a doxiciclina, > 4 µg/ml de valnemulina, 8 µg / mL para a tiamulina, 32 µg / ml para tilvalosina, > 64 µg / ml para a lincomicina e > 128 µg / ml de tilosina. Estes resultados corroboram em grande parte com os relatados na literatura. Tiamulina, doxiciclina e tilvalosina apresentaram os menores resultados de MIC. Todas as amostras submetidas à análise filogenética com base na sequência do gene nox exibiram resultados semelhantes, indicando 100% de identidade com B. hyodysenteriae. Este é o primeiro estudo que descreve o padrão MIC de B. hyodysenteriae isoladas no Brasil.(AU)

Malignant peripheral nerve sheath tumour in a sow.

Nodular lung lesions in swine are frequently due to abscesses or granulomatous pneumonia. Although tumours are rarely reported in modern pig farming, they should be considered as a differential diagnosis when nodular lung lesions are found. A first-parity sow exhibiting respiratory signs was euthanized. Several whitish firm nodules, not encapsulated, ranging in diameter from 0.5 to 5 cm were present in all lung lobes. Microscopically, the nodules were composed of dense neoplastic cells, mainly in Antoni types A and B patterns, infiltrative and with development of emboli. All neoplastic cells stained positively by immunohistochemistry for vimentin and S-100 protein, with variable immunostaining for glial fibrillary acidic protein and stained negative for cytokeratin. Based on the gross, histological and immunohistochemical features, the tumor was diagnosed as malignant peripheral nerve sheath tumour.