Evaluation of Mycobacterium avium subsp. paratuberculosis faecal culture protocols and media

Paratuberculosis is an important enteritis of ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). The disease is officially considered exotic in Brazil, but recent serological surveys and the isolation of the agent suggest it may occur in our herds. The aim of this study was to evaluate three different formulations of Herrold's egg yolk agar with mycobactin J (HEYM) and four faecal culture protocols considering their ability for Map growth as well as cost and ease of application. Three formulations of HEYM were inoculated with two suspensions of Map. Spiked faeces and naturally contaminated faecal samples were treated by the four faecal culture protocols. Centrifugation protocol and HEYM recommended by OIE showed the best results on the recovery of Map.

A seroepidemiological survey of Mycobacterium avium subsp. paratuberculosis in sheep from North of Portugal

The aim of this study was evaluate the risk factors for Mycobacterium avium subsp. paratuberculosis (Map) seroprevalence in sheep in the North of Portugal. The effects on seroprevalence of several variables such as individual characteristics, management practices, farm characteristics, animal health, and available veterinary services were evaluated. This information was then used in a multivariable logistic regression model in order to identify risk factors for Map seropositivity. Univariable analysis was used to screen the variables used in the logistic regression model. Variables that showed p values of <0.15 were retained for the multivariable analysis. Fifteen variables were associated with paratuberculosis in univariable analysis. The multivariable logistic regression model identified a number of variables as risk factors for seropositivity like sheep pure local and/or a cross of a local breed (OR=2.02), herd size with 31-60 head (OR=2.14), culling during the Spring-Summer season (OR=1.69) and the use of an anti-parasitic treatment such as Ivermectin as the only anti-parasitic medication (OR=5.60). Potential risk factors identified in this study support current recommendations for the control of paratuberculosis.

Avaliação sorológica e de fatores de risco para a infecção por Mycobacterium avium subsp. paratuberculosis em rebanhos leiteiros da Microrregião de Garanhuns, Pernambuco

Objetivou-se com esse trabalho realizar um inquérito epidemiológico da infecção por Mycobacterium avium subsp. paratuberculosis (MAP) em bovinos leiteiros da microrregião de Garanhuns, Pernambuco, Brasil. Para este estudo foram coletadas amostras sanguíneas de 408 animais, provenientes de 19 rebanhos localizados em 15 municípios. O exame sorológico foi realizado por Ensaio Imunoenzimático (ELISA) indireto para detecção de anticorpos frente ao MAP. Em todas as propriedades, foi aplicado um questionário investigativo para análise dos fatores de risco, e as coordenadas geográficas coletadas por um aparelho de Global Position System (GPS) para realização da distribuição espacial. A prevalência da infecção por MAP foi de 2,7% (11/408; I.C. 1,4-4,9). O número de focos foi 47,4% (9/19). Na análise de regressão logística foi identificado como fator de risco a taxa anual de nascimentos superior a 51 bezerros/ano (OR 3,8; I.C. 1,1-13,1). Desta forma, conclui-se que a infecção por MAP encontra-se presente nos rebanhos bovinos leiteiros da microrregião estudada e que medidas de controle baseadas nos fatores de risco identificados devem ser implementadas com o objetivo de reduzir o número de focos da infecção.

Comparação de duas técnicas de isolamento do Mycobacterium avium subsp. paratuberculosis em amostras de fezes de ovinos com suspeita clínica de paratuberculose

A paratuberculose é uma enterite crônica granulomatosa causada por Mycobacterium avium subsp. paratuberculosis que afeta principalmente os ruminantes. A cultura de bactérias a partir de amostras de fezes e tecidos constitui um dos métodos mais eficazes de diagnóstico, sendo ainda o único método disponível para obtenção de isolamentos e estirpes de micobactérias. Contudo, este método apresenta baixa sensibilidade e requer meses de incubação antes do crescimento de colônias. Neste estudo, utilizou-se a cultura fecal como método de diagnóstico em ovinos de diferentes raças portuguesas, com sinais compatíveis com a doença. Fez-se ainda a comparação entre os meios de cultura Löwenstein Jensen® com micobactina® J e o de Middlebrook® 7H11 com OADC®, utilizados no isolamento da bactéria. As percentagens de isolamento em cada um os meios foram de 2,0% (6/300) para Löwenstein Jensen® com micobactina J e 1,0% (3/300) para Middlebrook® 7H11/OADC. As três amostras positivas no meio de Middlebrook® 7H11/OADC também foram positivas no meio de Löwenstein Jensen® com micobactina J e nenhuma foi somente positiva no meio de Middlebrook® 7H11/OADC. Os resultados deste estudo sugerem que o meio de Löwenstein-Jensen® com micobactina® J é mais efetivo para a obtenção de estirpes ovinas em Portugal.

Cytokine gene expression and molecular detection of Mycobacterium avium subspecies paratuberculosisin organs of experimentally infected mice

AbstractMycobacterium avium subspecies paratuberculosis (MAP) can infect ruminants and remain subclinical for long periods within herds. The identification of organs that are more susceptible to infection and the evaluation of cytokine expression at the site of infection are important to understand the pathogenesis of MAP. In this study, the probability of detection of MAP-DNA and the expression of cytokines in organs of C57BL/6 mice infected intraperitoneally for 120 days were evaluated. Among the evaluated organs, the spleen (85%), colon (75%) and liver (60%) had the highest frequency of positivity. When compared these frequencies between organs, it has been found that the spleen had 1.54 times as likely to be positive in relation to the ileum, and 2.0 times more likely in relation to the Peyer's patches. In addition, at 60 days post-infection, the spleen and the liver were responsible for upregulation of IFN-γ , and the ileum by TNF-α and IL-4. The results indicate that the spleen is the best organ for evaluating an experimental infection by MAP, especially in the initial stages of the infection. Moreover, it showed that the spleen, liver and ileum have a direct role in the inflammatory response in experimental models.

Effect of lipoarabinomannan from Mycobacterium avium subsp avium in Freund’s incomplete adjuvant on the immune response of cattle

The aim of the present study was to determine whether lipoarabinomannan (LAM), in combination with Freund’s incomplete adjuvant (FIA), was able to improve cell-mediated and antibody-mediated immune responses against ovalbumin (OVA) in cattle. Twenty-three calves were assigned to four treatment groups, which were subcutaneously immunized with either OVA plus FIA, OVA plus FIA and LAM from Mycobacterium avium subsp avium, FIA plus LAM, or FIA alone. Lymphoproliferation, IFN-γ production and cell subpopulations on peripheral blood mononuclear cells before and 15 days after treatment were evaluated. Delayed hypersensitivity was evaluated on day 57. Specific humoral immune response was measured by ELISA. Inoculation with LAM induced higher levels of lymphoproliferation and IFN-γ production in response to ConA and OVA (P < 0.05). Specific antibody titers were similar in both OVA-immunized groups. Interestingly, our results showed that the use of LAM in vaccine preparations improved specific cell immune response evaluated by lymphoproliferation and IFN-γ production by at least 50 and 25%, respectively, in cattle without interfering with tuberculosis and paratuberculosis diagnosis.

Isolamento de cepas de Mycobacterium avium em búfalos abatidos para consumo

Duas cepas micobacterianas, isoladas no parênquima pulmonar e linfonodo apical de búfalos abatidos para consumo, procedentes de criatórios localizados na Ilha de Marajó (PA) e submetidas à identificação segundo ensaios recomendados para o gênero Mycobacterium, foram identificadas como pertencentes ao complexo Mycobacterium avium. Apresentaram-se considerações relativas à associação desses organismos com a Aids -- e o papel dos alimentos nessa associação --, discutindo-se o impacto que a condição de germes oportunistas das espécies desse complexo têm na pandemia do HIV, assim como o risco potencial representado pelas infecções produzidas nos animais.

Mycobacterium avium complex (MAC): an unusual potential pathogen in cerebrospinal fluid of AIDS patients

Mycobacterium avium complex (MAC) is frequently isolated from patients with late complications of Acquired Immunodeficiency Syndrome (AIDS), especially in North America and Europe. However, its isolation from the central nervous system (CNS) has been seldom reported in these countries. MAC infections in AIDS patients in African and Latin American countries are believed to be uncommon. We report the isolation of MAC from cerebrospinal fluid (CSF) of 11 AIDS patients out of 1723 (0.63%) seen at "Centro de Referência e Treinamento - AIDS", São Paulo and discuss the significance of its isolation.

Mycobacterium avium complex (MAC) isolated from AIDS patients and the criteria required for its implication in disease

Before the AIDS pandemia, the Mycobacterium avium complex (MAC) was responsible in most cases for the pneumopathies that attack patients with basic chronic pulmonary diseases such as emphysema and chronic bronchitis36. In 1981, with the advent of the acquired immunodeficiency syndrome (AIDS), MAC started to represent one of the most frequent bacterial diseases among AIDS patients, with the disseminated form of the disease being the major clinical manifestation of the infection8. Between January 1989 and February 1991, the Section of Mycobacteria of the Adolfo Lutz Institute, São Paulo, isolated MAC from 103 patients by culturing different sterile and no-sterile processed specimens collected from 2304 patients seen at the AIDS Reference and Training Center and/or Emilio Ribas Infectology Institute. Disseminated disease was diagnosed in 29 of those patients on the basis of MAC isolation from blood and/or bone marrow aspirate. The other 74 patients were divided into categories highly (5), moderately (26) and little suggestive of disease (43) according to the criteria of DAVIDSON (1989)10. The various criteria for MAC isolation from sterile and non-sterile specimens are discussed.

Clinical and laboratory findings of disseminated Mycobacterium avium complex infection (DMAC) in a pair matched case-control study

A pair matched case/control study was conducted from January 1991 to 30 June 1992 in order to define clinical and laboratory findings associated with DMAC infection in AIDS patients. Since DMAC infection is usually associated with advanced immunodeficiency, and therefore also with other opportunistic illnesses, in addition to the number of CD4+ lymphocytes, cases and controls were matched using the following criteria: date of AIDS diagnosis and antiretroviral therapy, number and severity of associated opportunistic infections and, whenever possible, type of Pneumocystis carinii prophylaxis, age and gender, in this order of relevance. Cases (defined as patients presenting at least one positive culture for MAC at a normally sterile site) and controls presented CD4+ lymphocyte counts below 50 cel/mm3. A significantly higher prevalence of general, digestive and respiratory signs, increased LDH levels, low hemoglobin levels and CD4+ cell counts were recorded for cases when compared to controls. Increases in gGT and alkaline phosphatase levels seen in cases were also recorded for controls. In conclusion, the strategy we used for selecting controls allowed us to detect laboratory findings associated to DMAC infection not found in other advanced immunossupressed AIDS patients without DMAC.

Soft tissue abscess and lymphadenitis due to Mycobacterium avium Complex as an expression of immune reconstitution inflammatory syndrome after a second scheme of highly active antiretroviral therapy

Immune reconstitution inflammatory syndrome (IRIS) is an atypical and unexpected reaction related to highly active antiretroviral therapy (HAART) in human immunodeficiency virus (HIV) infected patients. IRIS includes an atypical response to an opportunistic pathogen (generally Mycobacterium tuberculosis, Mycobacterium avium complex, cytomegalovirus and herpes varicella-zoster), in patients responding to HAART with a reduction of plasma viral load and evidence of immune restoration based on increase of CD4+ T-cell count. We reported a case of a patient with AIDS which, after a first failure of HAART, developed a subcutaneous abscess and supraclavicular lymphadenitis as an expression of IRIS due to Mycobacterium avium complex after starting a second scheme of HAART.

Mediastinitis and pericardial effusion in a patient with AIDS and disseminated Mycobacterium avium infection: a case report

We report the case of a 36-year-old man who had acquired immune deficiency syndrome and developed suppurative mediastinitis extending over the left lung and anterior thoracic wall around the sternum, pericardial effusions, splenomegaly, and mesenteric and periaortic lymphadenomegaly due to Mycobacterium avium (genotype I). The organism was isolated from an axillary lymph node and the bone marrow. Mediastinitis associated with disseminated M. avium complex infection is uncommon and, to the best of our knowledge, this manifestation has not reported before.

Analysis of Mycobacterium avium Complex Serovars Isolated from AIDS Patients from Southeast Brazil

The purpose of this study was to assess the distribution of Mycobacterium avium serovars isolated from AIDS patients in São Paulo and Rio de Janeiro. Ninety single site or multiple site isolates from 75 patients were examined. The most frequent serovars found were 8 (39.2%), 4 (21.4%) and 1 (10.7%). The frequency of mixed infections with serovar 8 or 4 was 37.8%. Among the 90 strains examined, M. intracellulare serovars (7 strains) and M. scrofulaceum (4 strains) were found in 11 isolates (12%) indicating that M. avium (88%) was the major opportunistic species in the M. avium complex isolates in Brazilian AIDS patients