Escherichia coli in Chicken Carcasses in Southern Brazil: Absence of Shigatoxigenic (STEC) and Isolation of Atypical Enteropathogenic (aEPEC)

The aim of this study was to investigate the presence of shiga toxin-producing Escherichia coli (STEC) and atypical enteropathogenic Escherichia coli (aEPEC) in frozen chicken carcasses sold at stores in southern Brazil. Typical E. coli colonies were enumerated in 246 chicken carcasses, and the presence of stx1, stx2, eae genes was investigated in their rinse liquid and in E. coli strains isolated from those carcasses. Strains of E. coli were also investigated for the presence of bfp gene. A median of 0.6 cfu.g-1(ranging from 0.1 to 242.7 cfu.g-1) of typical E. coli colonies was found in the carcasses. Shiga toxin-encoding genes (stx1 and stx2) were not detected, indicating that the chicken carcasses were negative for STEC. The intimin protein gene (eae) was detected in E.coli isolated from 4.88% of the carcasses; all tested strains were negative for the bfp gene and were classified as aEPEC. Twenty-two aEPEC strains were tested for resistance to ten antimicrobials and subjected to macrorestriction (PFGE). All the tested aEPEC strains were fully susceptible to cephalosporins, ciprofloxacin and colistin. Resistance to sulfonamide (65%), ampicillin (55%), tetracycline (50%) and gentamicin (45%) were the most frequent. The PFGE profile demonstrated a low level of similarity among the resistant strains, indicating that they were epidemiologically unrelated. The results indicate that aEPEC strains can contaminate chicken meat, and their association with strains implicated in human diarrhea needs to be further investigated.(AU)

New Isospora and Host Species in Brazilian Passerines

Normal passerines (n=216) were evaluated for oocysts of Isospora in feces at the Triage Center for Wild Animals (CETAS, IBAMA, Belo Horizonte; August 21 to September 21, 2012). The positive samples with oocysts represented 13.0% of Cardinalidae (n=23), 11.2% of Emberizidae (n=107), 50% of Icteridae (n=10) and 60.3% of Thraupidae (n=68). The probability of fecal oocysts attributable to the host in Thraupidae is higher than in Cardinalidae, Emberizidae, Fringillidae and Turdidae, but similar to Icteridae. No oocysts were found in Fringillidae and Turdidae. Within Thraupidae, Isospora was for the first time described in Paroaria dominicana and Schistochlamys ruficapillus and within Icteridae, in Gnorimopsar chopi. Saltator similis presented a higher risk, 66.9% greater than Lanio pileatus and Sporophila caerulescens and with a 27.9% greater probability than Sporophila nigricolis. The new coccidian species described were Isospora dominicana [ellipsoid oocysts, 25 (30-20) x 25 (28-20) µm] in Paroaria dominicana; Isospora beagai [ovoid oocysts, 28 (32-17) x 25 (29-16) µm] and Isospora ferri [ellipsoid oocysts, 20 (22-16) x 18 (22-15) µm] in Saltator similis; Isospora ruficapillus [spheric to subspherical oocysts, 25 (26-23) x 24 (25-21) µm] in Schistochlamys ruficapillus; and Isospora chopi [spherical to sub-spherical oocysts, 24.5 x 22 (30-20 x 25-20) µm] and Isospora gnorimopsar [sub-spherical to ovoid oocysts, 27 x 23 (32-22 x 28-20) µm] in Gnorimopsar chopi. The morphometry and features were compared with previously described Isospora in passerines. New coccidian species and new passerine hosts are described for Isospora and recommends for constant monitoring during rehabilitation, especially for the hosts of Thraupidae and Icteridae.(AU)

New Isospora and Host Species in Brazilian Passerines

Normal passerines (n=216) were evaluated for oocysts of Isospora in feces at the Triage Center for Wild Animals (CETAS, IBAMA, Belo Horizonte; August 21 to September 21, 2012). The positive samples with oocysts represented 13.0% of Cardinalidae (n=23), 11.2% of Emberizidae (n=107), 50% of Icteridae (n=10) and 60.3% of Thraupidae (n=68). The probability of fecal oocysts attributable to the host in Thraupidae is higher than in Cardinalidae, Emberizidae, Fringillidae and Turdidae, but similar to Icteridae. No oocysts were found in Fringillidae and Turdidae. Within Thraupidae, Isospora was for the first time described in Paroaria dominicana and Schistochlamys ruficapillus and within Icteridae, in Gnorimopsar chopi. Saltator similis presented a higher risk, 66.9% greater than Lanio pileatus and Sporophila caerulescens and with a 27.9% greater probability than Sporophila nigricolis. The new coccidian species described were Isospora dominicana [ellipsoid oocysts, 25 (30-20) x 25 (28-20) µm] in Paroaria dominicana; Isospora beagai [ovoid oocysts, 28 (32-17) x 25 (29-16) µm] and Isospora ferri [ellipsoid oocysts, 20 (22-16) x 18 (22-15) µm] in Saltator similis; Isospora ruficapillus [spheric to subspherical oocysts, 25 (26-23) x 24 (25-21) µm] in Schistochlamys ruficapillus; and Isospora chopi [spherical to sub-spherical oocysts, 24.5 x 22 (30-20 x 25-20) µm] and Isospora gnorimopsar [sub-spherical to ovoid oocysts, 27 x 23 (32-22 x 28-20) µm] in Gnorimopsar chopi. The morphometry and features were compared with previously described Isospora in passerines. New coccidian species and new passerine hosts are described for Isospora and recommends for constant monitoring during rehabilitation, especially for the hosts of Thraupidae and Icteridae.

Escherichia coli in Chicken Carcasses in Southern Brazil: Absence of Shigatoxigenic (STEC) and Isolation of Atypical Enteropathogenic (aEPEC)

The aim of this study was to investigate the presence of shiga toxin-producing Escherichia coli (STEC) and atypical enteropathogenic Escherichia coli (aEPEC) in frozen chicken carcasses sold at stores in southern Brazil. Typical E. coli colonies were enumerated in 246 chicken carcasses, and the presence of stx1, stx2, eae genes was investigated in their rinse liquid and in E. coli strains isolated from those carcasses. Strains of E. coli were also investigated for the presence of bfp gene. A median of 0.6 cfu.g-1(ranging from 0.1 to 242.7 cfu.g-1) of typical E. coli colonies was found in the carcasses. Shiga toxin-encoding genes (stx1 and stx2) were not detected, indicating that the chicken carcasses were negative for STEC. The intimin protein gene (eae) was detected in E.coli isolated from 4.88% of the carcasses; all tested strains were negative for the bfp gene and were classified as aEPEC. Twenty-two aEPEC strains were tested for resistance to ten antimicrobials and subjected to macrorestriction (PFGE). All the tested aEPEC strains were fully susceptible to cephalosporins, ciprofloxacin and colistin. Resistance to sulfonamide (65%), ampicillin (55%), tetracycline (50%) and gentamicin (45%) were the most frequent. The PFGE profile demonstrated a low level of similarity among the resistant strains, indicating that they were epidemiologically unrelated. The results indicate that aEPEC strains can contaminate chicken meat, and their association with strains implicated in human diarrhea needs to be further investigated.

Morphological and molecular characterization of Haemoproteus coatneyi and Haemoproteus erythrogravidus (Haemosporida: Haemoproteidae) in Passeriformes in Brazils Atlantic Forest / Caracterização morfológica e molecular de Haemoproteus coatneyi e Haemoproteus erythrogravidus (Haemosporida: Haemoproteidae) em Passeriformes da Mata Atlântica, Brasil

Haemoproteus spp. are protozoan parasites found in birds around the world. These parasites are identified through the morphology of gametocytes, phylogenetic analysis based on the mitochondrial cytb gene, and the parasites geographic distribution. The absence of erythrocytic merogony, high intraspecific genetic variation and low parasitemia in wild birds makes it essential to use integrative approaches that assist in the identification of these parasites. Thus, microscopic and molecular analyses, combined with spatial distribution, were carried out to verify the presence of Haemoproteus spp. in wild birds in Brazil. Light microscopy revealed one Tangara sayaca bird was parasitized by Haemoproteus coatneyi and, two specimens of Zonotrichia capensis presented Haemoproteus erythrogravidus. The morphology of the gametocytes of these two parasitic species showed high similarity. The molecular analysis revealed the presence of one lineage of H. coatneyi and two lineages of H. erythrogravidus, one of which is considered a new lineage. These lineages were grouped phylogenetically in separate clades, with low genetic divergence, and the H. erythrogravidus lineage emerged as an internal group of the lineages of H. coatneyi. The geographic distribution demonstrated that the two species occur in the American continent. This is the first report of H. erythrogravidus in Brazil.(AU)

Haemoproteus spp. são protozoários parasitos encontrados em aves de todo o mundo. A identificação desses parasitos é realizada por meio da morfologia dos gametócitos, da análise filogenética, baseada no gene mitoncodrial cytb e na distribuição geográfica do parasito. A ausência de merogonia eritrocítica, a alta variação genética intraespecífica e a baixa parasitemia em aves silvestres, tornam essencial a utilização de abordagens integrativas que auxiliem na identificação desses parasitos. Assim, análises microscópicas e moleculares, aliadas à distribuição espacial, foram realizadas para verificar a presença de Haemoproteus spp. em aves silvestres no Brasil. A microscopia óptica demonstrou que uma ave Tangara sayaca estava parasitada por Haemoproteus coatneyi, e dois espécimes de Zonotrichia capensis apresentavam Haemoproteus erythrogravidus, cujas morfologias dos gametócitos apresentaram alta similaridade. A análise molecular recuperou uma linhagem de H. coatneyi e duas linhagens de H. erythrogravidus, sendo uma dessas considerada nova linhagem. Essas linhagens se agruparam filogeneticamente em clados separados, apresentando baixa divergência genética, sendo que as linhagens de H. erythrogravidus emergiram como grupo interno às linhagens de H. coatneyi. A distribuição geográfica demonstrou que as duas espécies estão ocorrendo no continente americano. Este é o primeiro relato de H. erythrogravidus no Brasil.(AU)

Research on Cryptococcus neoformans and Candida spp. in the excretas of captive psittacisittacines and passeriformes / Pesquisa de Cryptococcus neoformans e Candida spp. em excretas de psitacídeos e passeriformes cativos / Investigación de Cryptococcus neoformans y Candida spp. en excretas de psittacidae y psittaciformes cautivos

As leveduras podem causar diversas doenças no homem e animais. Nas aves, as leveduras estão envolvidas principalmente em lesões no trato respiratório e digestório. Entre as leveduras patogênicas, Cryptococcus neoformans vem se destacando pela alta prevalência de criptococose humana em pacientes imunodeprimidos. Assim, os objetivos deste estudo foram identificar C. neoformans e outras leveduras patogênicas na cloaca e coana de passeriformes e psitaciformes e em excretas coletadas do fundo de gaiolas de aviários. Foram obtidas 29 amostras de 15 aves manifestando algum sinal respiratório, provenientes do Ambulatório de Animais Selvagens da UFPR (n=6) e da Clínica Veterinária Vida Livre (n= 23). As amostras foram semeadas em Ágar Sabouraud e Ágar Níger e mantidas a 300 C por até 30 dias. Todas as colônias foram analisadas quanto à macro e micromorfologia. Para aquelas identificadas como leveduras, foram realizadas as provas bioquímicas: assimilação de carbono e nitrogênio e formação de tubo germinativo para identificação de Candida albicans. As amostras de excreta dos aviários (n=8) foram misturadas com solução fisiológica contendo antibiótico e o sobrenadante foi semeado em Ágar Níger. Nenhuma amostra das aves apresentou resultado positivo para C. neoformans, porém identificaram-se amostras positivas para C. albicans (duas amostras de coana), C. famata (uma amostra de coana) e C. tropicalis (uma amostra de coana). As excretas foram negativas para C. neoformans. Portanto, apesar de não ter sido isolado C. neoformans, outras leveduras patogênicas foram isoladas, demonstrando a importância dessas aves como possíveis veiculadoras de doenças para humanos.(AU)

The yeasts can cause many diseases in man and animals. On birds, the yeasts are involved mainly in respiratory and digestive tract lesions. Among pathogenic yeast, Cryptococcus neoformans is an important cause of human cryptococcisis associated with immunocompromised states. The purpose of this study is to identify the occurrence of C. neoformans and other pathogenic yeasts in cloacae and choana from passeriformes and psittacines as well as in excretas from poultry cages. Twenty nine samples from fifteen birds showing some respiratory symptom, from Veterinary Hospital of UFPR (n = 6) and Vida Livre Veterinary Clinic (n = 23), were collected . The samples were spread in Sabouraud dextrose Agar and Staib medium and kept at 30°C and observed for 30 days. All colonies were analyzed with respect to its micro and macromorphology. Biochemical assays were conducted for samples presenting yeasts: carbon and nitrogen assimilation profile and germ tube for Candida albicans identification. Samples from birdsextracts (n = 8) were diluted in sterile saline solution with antibiotic and the supernatant was inoculated in spread on Niger seed agar. All samples were negative for Cryptococcus neoformans, however, C. albicans (two samples from choana), C. famata (one sample from choana) and C. tropicalis (choana) were found. Excretas from bird cages were negative to C. neoformans. Results suggested that birds harbor various pathogenic species of yeast, but not C. neoformans, and the result showed potential danger to carry diseases to humans.(AU)

Las levaduras pueden causar diversas enfermedades en el hombre y animales. En las aves, las levaduras están involucradas principalmente en lesiones en el tracto respiratorio y digestivo. Entre las levaduras patogénicas, Cryptococcus neoformans viene destacándose por la alta incidencia de cryptococcus humana en pacientes inmune deprimidos. Así, el objetivo de este estudio fueron identificar C. neoformans y otras levaduras patogénicas en la cloaca y coana de psittacidae y psittaciformes y en excretas colectadas de las jaulas de pajareras. Fueron obtenidas 29 muestras de quince (15) aves manifestando algún señal respiratorio, provenientes del Ambulatorio de Animales Salvajes de la UFPR (n=6) y de la Clínica Veterinaria Vida Livre (n= 23). Las muestras fueron sembradas en Ágar Sabouraud y Ágar Níger y mantenidas a 30 0 C hasta 30 días. Todas las colonias fueron analizadas cuanto a la macro y micromorfología. Para aquellas identificadas como levaduras, fueron realizadas las pruebas bioquímicas: asimilación de carbono y nitrógeno, y formación de tubo germinativo para identificación de Candida albicans. Las muestras de excreta de los pajareros (n=8) fueron mezcladas con solución fisiológica conteniendo antibiótico y el sobrenadante fue sembrado en Ágar Níger. Ninguna muestra de las aves presentó resultado positivo para C. neoformans, pero se identificaron muestras positivas para C. albicans (dos muestras de coana), C. famata (una muestra de coana) y C. tropicalis (una nuestra de coana). Las excretas fueron negativas para C. neoformans. Por lo tanto, a pesar de no haber sido aislado C. neoformans, otras levaduras patogénicas fueron aisladas, demostrando que esas aves son posibles transmisoras de enfermedades para los seres humanos.(AU)