Comparative analysis of diagnostic methods and risk factors for Aelurostrongylus abstrusus infection in brazilian cats.

This study aimed to prospectively evaluate the risk factors of infection by Aelurostrongylus abstrusus in Brazilian cats with cough and/or radiographic changes, using as diagnostic tools the Baermann method (BM), polymerase chain reaction (PCR) of feces, bronchoalveolar lavage fluid (BALF), and cytology. Forty-three cats that were presented with cough or lung radiographic abnormalities compatible with bronchoalveolar disease were included in the study. After clinical evaluation, feces samples were collected to investigate lungworm parasitism through BM and PCR. BALF was performed to provide samples for cytology, bacteriology, and fungal culture. Stool PCR was considered the gold standard for diagnosis tests, and the other methods were evaluated by their agreement. PCR presented 74% (32/43) of positivity for A. abstrusus, while in the BM, 41% (18/43) were positive. BM showed sensitivity of 56.25% and specificity of 100% when compared with PCR. No larva was found in the cytological evaluation of 21 BALF samples. Lungworm is an important cause of bronchopulmonary disease in domestic cats in Brazil and should be included as a differential diagnosis when a cat is presented with cough or radiographic abnormalities. BM is a sensitive, non-invasive, and cheap technique to diagnose the disease, but it is not as sensitive as PCR.

Necrotizing Tracheobronchitis Caused by Aspergillus fumigatus in a Cow.

This report describes a case of tracheobronchitis caused by Aspergillus fumigatus in a cow. A 4-year-old Jersey cow was submitted for necropsy examination. Grossly, the mucosa of the trachea and bronchi was diffusely reddened and was covered by a moderate amount of yellow, caseous material, sometimes with small foci of grey 'cotton-like' appearance, suggestive of filamentous fungus. Microscopical analysis of the trachea and bronchi showed marked diffuse mucosal and submucosal necrosis with a large amount of cellular debris, fibrinous exudation and inflammatory infiltration dominated by macrophages and neutrophils. There were large numbers of septate fungal hyphae with acute bifurcation angles and conidia and vesicles typical of Aspergillus spp. that were stained by the Grocott method. A. fumigatus growth in mycological culture from the trachea and bronchi was confirmed by molecular identification. Tracheobronchitis caused by A. fumigatus infection in cattle is poorly described and requires further research to improve the therapeutic management of the disorder.

Chronic Invasive Pulmonary Aspergillosis in Two Cats with Diabetes Mellitus.

Human patients with diabetes mellitus are at increased risk of fungal infections. Diabetes mellitus has also been implicated as a predisposing factor in the establishment of fungal lung infections in cats. Two diabetic cats of different origins presented with severe acute respiratory conditions that resulted in their death. At necropsy examination there was friable, black material in the main bronchi that obstructed the bronchial lumina. Microscopical examination of the lungs revealed the presence of pneumonia, calcium oxalate crystals and a large quantity of fungal hyphae and conidial heads. Fungal infection was confirmed with Grocott's methenamine silver stain. The results of the mycology analysis were compatible with Aspergillus section Nigri.

Pneumocystis sp. in bats evaluated by qPCR.

Molecular techniques have revealed a high prevalence of Pneumocystis colonization in wild mammals. Accurate quantification of Pneumocystis sp. is essential for the correct interpretation of many research experiments investigating this organism. The objectives of this study were to detect the presence of Pneumocystis sp. in bats by qPCR, and to distinguish colonization from infection. Probes and primers for real time PCR (qPCR) were designed based on the gene of major surface glycoprotein (MSG) of Pneumocystis sp., in order to analyze 195 lung tissue samples from bats captured (2007-2009). All samples were also analyzed by nested PCR, using oligonucleotide primers designed for the gene encoding the mitochondrial small subunit rRNA (mtSSU rRNA) to confirm the results. The qPCR assay was standardized using a standard curve made with the DNA extracted from bronchoalveolar lavage positive for Pneumocystis jirovecii. The average Ct was found to be between 13 and 14 (calibration curve) for the detection of infection with Pneumocystis sp. and above these values for colonization. It was considered as negative samples the ones that had Ct values equal to 50. Out of the total 195 samples, 47 (24.1%) bat lung DNA samples were positive for Pneumocystis sp. by qPCR. The most common bat species found were: Tadarida brasiliensis (23.4%), Histiotus velatus (17.0%), Desmodus rotundus (14.9%) and Molossus molossus (8.5%). The average cycle threshold of the positive samples (bats) was 25.8 and standard deviation was 1.7. The DNA samples with Ct values greater than 14 suggest that these animals might be colonized by Pneumocystis sp. Results obtained in this study demonstrated the usefulness of the qPCR procedure for identification of Pneumocystis sp. and for distinction between its colonizing or infectious status in bats.

Phylogenetic analysis of Pneumocystis from pig lungs obtained from slaughterhouses in southern and midwestern regions of Brazil / Análise filogenética de Pneumocystis obtidos de pulmões de suínos provenientes de abatedouros da região sul e centro oeste do Brasil

The Pneumocystis genus is comprised of pathogens dwelling in the lungs of terrestrial, aerial, and aquatic mammals. Occasionally they induce severe pneumonitis, particularly in hosts with severe impairment of the immune system and progressively may fill pulmonary alveolar cavities causing respiratory failure. Molecular genetic studies revealed that Pneumocystis gene sequences present a marked divergence with the host species concerned. In the present study, the genetic diversity of Pneumocystis obtained from lungs of swines was examined by analyzing mitochondrial large subunit (mtLSU) and small subunit (mtSSU) rRNA sequences. The samples were obtained from two slaughterhouses located in two Brazilian states. Phylogenetic analysis demonstrated that genetic groupings within Pneumocystis organisms were in accordance with those of the corresponding hosts and that two clusters were formed. In conclusion, these data show that there are genetically distinct porcine Pneumocystis genotypes with at least two separate clusters in Brazil.

O gênero Pneumocystis compreende patógenos que residem em pulmões de animais terrestres, aéreos e aquáticos. Pode ocasionar uma grave pneumonia, particularmente em hospedeiros com o sistema imunológico seriamente comprometido, o que ocorre por meio de uma progressiva disseminação nas cavidades alveolares, causando insuficiência respiratória. Estudos genéticos, baseados em métodos moleculares, revelaram que as sequências dos genes de Pneumocystis apresentam marcante divergência de acordo com a espécie de hospedeiro. Neste estudo, a diversidade genética das amostras obtidas a partir de pulmões de suínos, provenientes de dois abatedouros localizados em dois estados brasileiros, foi examinada por análise das sequencias dos nucleotídeos dos produtos de PCR dos genes mtLSU e mtSSU do rRNA do Pneumocystis. O resultado confirma a tendência registrada em pesquisas com amostras de outros animais e permite concluir que existem, pelo menos, dois grupos filogenéticos distintos de Pneumocystis de suínos no Brasil.

Detection of Pneumocystis spp. in lung samples from pigs in Brazil.

The genus Pneumocystis is composed of opportunistic fungi currently considered as specific pulmonary pathogens in humans and other mammals. In pigs, Pneumocystis pneumonia (PcP) could create significant economical losses due to its detrimental effects on growth, food conversion, and carcass/viscera condemnation. This study revealed that Pneumocystis organisms could be detected by Grocott's staining or immunohistochemistry in 36.9% of 564 slaughtered pigs from two geographic regions of Brazil. The prevalence of positive cases was 39.9% and 33.9% in pigs slaughtered in Rio Grande do Sul (RS) and Mato Grosso (MT) states, respectively. Among the positive cases in RS, Pneumocystis organisms were observed in 41.9% of 33 histologically normal lungs, and in 58.0% of lungs presenting with histological lesions. In contrast, the prevalence in MT in normal and abnormal lungs was 36.3% and 63.5%, respectively. Major histopathological findings in lungs of infected animals were bronchointerstitial pneumonia (47.6%), suggestive of enzootic pneumonia, and interstitial pneumonia (37.9%), compatible with PcP. The results of this survey strengthened the interest of detecting fungal pathogens, in addition to other infectious agents, and evaluating their financial impact on Brazilian pig industry. Preventive and/or therapeutic strategies should be developed in order to minimize the incidence of respiratory fungal infections in pigs and associated economic losses.