Methicillin-resistant Staphylococcus pseudintermedius clonal groups isolated from canine pyoderma in Brazil

Background: Since the early reports of mecA-positive Staphylococcus (S.) pseudintermedius isolates in the United States and in Europe were published, the frequency of methicillin-resistant S. pseudintermedius (MRSP) has increased among skin disease cases in dogs in many countries. Moreover, MRSP isolates frequently present a multi-drug resistant profile, which include most drugs used for the skin disease treatment. The distribution of multi- drug resistant MRSP clonal groups in turn varies according to geographic region. Despite the large dog population in Brazil, no data on the MRSP resistance profile or clonal groups have been reported. The aim of this study was to assess the antimicrobial resistance phenotypes and clonal relationships of MRSP isolates originating from dogs affected by recurrent skin diseases.Material, Methods & Results: Twenty-one epidemiologically unrelated isolates originating from dogs inflicted with a recurrent skin disease, which were treated at the Veterinary Hospital (HCV) of the Federal University of Rio Grande do Sul (UFRGS) in Porto Alegre, were included in this study. The isolates suspected of being MRSP were subjected to PCR analysis to confirm their identity. Identifications were made using PCR analysis that targeted the mecA gene and PCRRFLP that targeted the pta gene. Isolates were further assessed by a disc diffusion test for resistance to 13 antimicrobials. Clonal groups were determined according to spa typing and SmaI fingerprinting (Pulsed-field Gel Electrophoresis-PFGE) profiles. All 21 isolates were confirmed to be MRSP and displayed a multiple resistance profile. In total, 4 different spa types were identified, and the most prevalent was a novel spa type (tyA) described in this study. SmaI-macrorestriction analysis demonstrated that the MRSP isolates presented between seven and twelve fragments and were distributed among 15 PFGE profiles.[...](AU)

Methicillin-resistant Staphylococcus pseudintermedius clonal groups isolated from canine pyoderma in Brazil

Background: Since the early reports of mecA-positive Staphylococcus (S.) pseudintermedius isolates in the United States and in Europe were published, the frequency of methicillin-resistant S. pseudintermedius (MRSP) has increased among skin disease cases in dogs in many countries. Moreover, MRSP isolates frequently present a multi-drug resistant profile, which include most drugs used for the skin disease treatment. The distribution of multi- drug resistant MRSP clonal groups in turn varies according to geographic region. Despite the large dog population in Brazil, no data on the MRSP resistance profile or clonal groups have been reported. The aim of this study was to assess the antimicrobial resistance phenotypes and clonal relationships of MRSP isolates originating from dogs affected by recurrent skin diseases.Material, Methods & Results: Twenty-one epidemiologically unrelated isolates originating from dogs inflicted with a recurrent skin disease, which were treated at the Veterinary Hospital (HCV) of the Federal University of Rio Grande do Sul (UFRGS) in Porto Alegre, were included in this study. The isolates suspected of being MRSP were subjected to PCR analysis to confirm their identity. Identifications were made using PCR analysis that targeted the mecA gene and PCRRFLP that targeted the pta gene. Isolates were further assessed by a disc diffusion test for resistance to 13 antimicrobials. Clonal groups were determined according to spa typing and SmaI fingerprinting (Pulsed-field Gel Electrophoresis-PFGE) profiles. All 21 isolates were confirmed to be MRSP and displayed a multiple resistance profile. In total, 4 different spa types were identified, and the most prevalent was a novel spa type (tyA) described in this study. SmaI-macrorestriction analysis demonstrated that the MRSP isolates presented between seven and twelve fragments and were distributed among 15 PFGE profiles.[...]

Identification of Brucella sp. isolated in Brazil from 1976 to 2013 by Bruce-Ladder PCR

Background: Brucella sp. are the causative agents of brucellosis, an infectious disease that affects various species ofanimals and can be transmitted to humans through direct contact with infected animals, indirectly by the ingestion of rawmilk products, and during the handling of strains or infected material in the laboratory. Being a zoonosis, the detectionof Brucella species in animals is essential for the prevention of the disease in humans and to perform a good program ofcontrol in infected herds. This study aimed at identifying Brucella field strains isolated from 1976 to 2013 in Brazil, usingthe modified Bruce-Ladder method, to evaluate the performance of this technique.Materials, Methods & Results: Eighty-three strains of Brucella sp. were included in the study, i.e. 21 reference strains(nine B. abortus, one B. canis, four B. melitensis, two B. ovis and five B. suis) and 62 field strains (six B. canis, oneB. suis and 55 B. abortus). For the identification of the genus and/or species of Brucella, biochemical and physiologicaltests, including MacConkey-agar growth, glucose fermentation, haemolysis, catalase, oxidase and urease tests, nitratereduction, citrate utilization, H2S production and CO2 requirement, were performed. Genomic DNA was extracted frompure cultures through heat-lysis of bacterial cultures and the genus was confirmed by a genus-specific PCR (bcsp31 targetgene), before performing the modified Bruce-Ladder PCR for the confirmation of the Brucella species. No problems ofspecificity were observed with the Bruce-Ladder PCR. However, the 1,682 bp fragment was not systematically amplified,even after several modifications such as the concentration of mix components, annealing temperatures and time. Therefore,an individual PCR using primers specific to this fragment was needed for complete identification of some strains. Also,only one kind of Polymerase gave the best results...(AU)

Identification of Brucella sp. isolated in Brazil from 1976 to 2013 by Bruce-Ladder PCR

Background: Brucella sp. are the causative agents of brucellosis, an infectious disease that affects various species ofanimals and can be transmitted to humans through direct contact with infected animals, indirectly by the ingestion of rawmilk products, and during the handling of strains or infected material in the laboratory. Being a zoonosis, the detectionof Brucella species in animals is essential for the prevention of the disease in humans and to perform a good program ofcontrol in infected herds. This study aimed at identifying Brucella field strains isolated from 1976 to 2013 in Brazil, usingthe modified Bruce-Ladder method, to evaluate the performance of this technique.Materials, Methods & Results: Eighty-three strains of Brucella sp. were included in the study, i.e. 21 reference strains(nine B. abortus, one B. canis, four B. melitensis, two B. ovis and five B. suis) and 62 field strains (six B. canis, oneB. suis and 55 B. abortus). For the identification of the genus and/or species of Brucella, biochemical and physiologicaltests, including MacConkey-agar growth, glucose fermentation, haemolysis, catalase, oxidase and urease tests, nitratereduction, citrate utilization, H2S production and CO2 requirement, were performed. Genomic DNA was extracted frompure cultures through heat-lysis of bacterial cultures and the genus was confirmed by a genus-specific PCR (bcsp31 targetgene), before performing the modified Bruce-Ladder PCR for the confirmation of the Brucella species. No problems ofspecificity were observed with the Bruce-Ladder PCR. However, the 1,682 bp fragment was not systematically amplified,even after several modifications such as the concentration of mix components, annealing temperatures and time. Therefore,an individual PCR using primers specific to this fragment was needed for complete identification of some strains. Also,only one kind of Polymerase gave the best results...

Diagnóstico imuno-histoquímico e caracterização anatomopatológica de clamidiose em psitacídeos / Immunohistochemical diagnosis and pathological characterization of chlamydiosis in psittacine

A clamidiose é causada por Chlamydophila psittaci e representa uma das principais zoonoses de origem aviária. Realizou-se um estudo retrospectivo em psitacídeos do período de 1995 a 2012 e exame imuno-histoquímico (IHQ) anti-Chlamydia. Foram avaliados 111 casos, dos quais 12 foram a óbito devido à clamidiose. As aves eram provenientes de apreensão ou cativeiro (zoológicos, criatórios, centros de triagem e domicílios). À necropsia observou-se fígado aumentado (4/12) com áreas branco-amareladas (3/12), baço aumentado (2/12) e rompido (1/12), saco pericárdico com deposição de fibrina (1/12), polisserosite fibrinosa (1/12) e em três casos não havia lesões. Na avaliação histopatológica evidenciou-se hepatite necrótica mononuclear (7/12), hepatite mononuclear (3/12), hiperplasia de ductos biliares (8/12), esplenite necrótica histiocitária (9/12), hemossiderose em fígado (9/12) e baço (9/12), aerossaculite mononuclear (4/12), pericardite fibrino-heterofílica (2/12), necrose (1/12) e rarefação (1/12) linfoide de bursa de Fabricius, pneumonia fibrinosa (1/12), nefrite mononuclear (1/12) e granulomas renais (1/12). Observaram-se inclusões basofílicas intracitoplasmáticas (corpos elementares) em fígado (2/12), baço e rins (1/12). Evidenciou-se imunomarcação anti-Chlamydia em fígado (11/12), baço (7/9), pulmões (3/9), rins (2/8), intestinos (2/3), sacos aéreos (1/4) e bursa de Fabricius (1/2). A IHQ poderá ser utilizada como forma de diagnóstico definitivo post mortem de clamidiose em psitacídeos no Brasil.(AU)

Chlamydiosis is caused by Chlamydophila psittaci and is one of the most important avian zoonosis. A retrospective study in psittacines was performed from 1995 to 2012 with immunohistochemistry (IHC) anti-Chlamydia. Hundred eleven cases were evaluated and twelve birds died due to chlamydiosis. The birds were obtained from illegal commerce traffic or captive conditions (zoos, breeding birds, wildlife rehabilitation center and pets). Grossly, there were hepatomegaly (4/12) with yellowish-white areas (3/12), splenomegaly (2/12), splenic rupture (1/12), fibrin deposition in pericardial sac (1/12), fibrinous polyserositis (1/12), and in three cases lesion was not found. Histopathological evaluation revealed mononuclear necrotizing hepatitis (7/12), mononuclear hepatitis (3/12), biliary duct hyperplasia (8/12), histiocytic necrotizing splenitis (9/12), hemosiderosis in liver (9/12) and spleen (9/12), mononuclear aerosaculitis (4/12), fibrin heterophilic pericarditis (2/12), lymphoid necrosis (1/12) and depletion of bursa Fabricius (1/12), fibrinous pneumonia (1/12), mononuclear nephritis (1/12), and renal granulomas (1/12). Basophilic intracytoplasmic inclusions (elementary bodies) were observed in liver (2/12), spleen and kidney (1/12). Positive immunostaining for Chlamydia could be detected in liver (11/12), spleen (7/9), lung (3/9), kidney (2/8), intestines (2/3), air sacs (1/4) and bursa of Fabricius (1/2). It was concluded that IHC can be used as postmortem definitive diagnosis of chlamydiosis in psittacines.(AU)