Occurrence of Apicomplexa protozoa in wild birds in the Northeast region of Brazil / Ocorrência de protozoários Apicomplexa em aves silvestres na região Nordeste do Brasil

Protozoa of the Apicomplexa phylum are worldwide distributed with capacity to infect endothermic animals. The study of these protozoa in wild birds in Brazil is scarce. This study aimed to evaluate the occurrence of apicomplexan protozoa in wild birds in the Northeast of Brazil. From October to December 2019, brain tissue samples were collected from 71 captive birds from the Wild Animal Screening Center of the Pernambuco State (CETRAS-Tangara) and 25 free-living birds from the Caatinga biome in Rio Grande do Norte, totaling 96 animals (41 species). Brain fragments were subjected to molecular diagnosis by nested PCR for the 18s rDNA gene of Apicomplexa parasites, followed by DNA sequencing. This gene was detected in 25% (24/96) of the samples, and it was possible to perform DNA sequencing of 14 samples, confirming three genera: Isospora, Sarcocystis and Toxoplasma from eight bird species (Amazona aestiva, Coereba flaveola, Egretta thula, Paroaria dominicana, Sporophila nigricollis, Cariama cristata, Columbina talpacoti, Crypturellus parvirostris). The occurrence these coccidia in wild birds provides important epidemiological information for the adoption of preventive measures for its conservation. Future studies are needed to better understand the consequence of Apicomplexa infection in birds in Caatinga and Atlantic Forest biomes.(AU)

Protozoários do filo Apicomplexa são distribuídos mundialmente e com capacidade de infectar animais endotérmicos. O estudo destes protozoários, em aves silvestres do Brasil, é escasso. Objetivou-se avaliar a ocorrência de protozoários Apicomplexa em aves silvestres na região Nordeste do Brasil. De outubro a dezembro de 2019, foram coletadas amostras de encéfalo de 71 aves de cativeiro do Centro de Triagem e Reabilitação de Animais Silvestres de Pernambuco (CETRAS-Tangara). E 25 aves de vida livre do bioma Caatinga no Rio Grande do Norte, totalizando 96 animais (41 espécies). Os fragmentos de encéfalo foram submetidos ao diagnóstico molecular por nested PCR, para o gene 18s rDNA de protozoários Apicomplexa, seguido por sequenciamento do DNA. Este gene foi detectado em 25% (24/96) das amostras analisadas; foi possível realizar o sequenciamento de 14 amostras, confirmando-se três gêneros: Isospora, Sarcocystis e Toxoplasma em oito espécies de aves (Amazona aestiva, Coereba flaveola, Egretta thula, Paroaria dominicana, Sporophila nigricollis, Cariama cristata, Columbina talpacoti, Crypturellus parvirostris). A ocorrência destes coccídios nas aves silvestres fornece informações epidemiológicas importantes para a adoção de medidas preventivas tendo em vista sua conservação. Estudos futuros são necessários para melhor compreensão da consequência da infecção por Apicomplexa, em aves silvestres dos biomas Caatinga e Floresta Atlântica.(AU)

Apicomplexa parasites in the brains of road-killed wild animals in the State of Paraná, Brazil / Parasitas apicomplexas em cérebro de animais silvestres mortos em estradas do Estado do Paraná, Brasil

The use of run-over wild animals is an efficient strategy for scientific research of pathogens. The aim of this study was to detect DNA from phylum Apicomplexa in the brain of road-killed wild animals from the NorthCentral and North Pioneer mesoregions of Paraná, Brazil. Pre-established transects were run weekly; when found, animals were packed into individual packages and sent for autopsy. The brain fragments were collected and kept at -20 ° C until processing. The DNA extracted from the samples was amplified by nested-PCR for the 18S rDNA gene from the phylum Apicomplexa. All positive samples were submitted to DNA sequencing to define the species. A total of 90 animals were collected, however, only 68 animals (75.6%) that had integrity of the brain were included in the study. It was possible to identify the species by DNA sequencing in four samples: Sarcocystis spp. was identified in one Colaptes melanochloros (Greenbarred woodpecker) and one Mazama gouazoubira (Gray brocket). Neospora caninum was observed in a Leopardus pardalis (Ocelot) and T. gondii was present in Didelphis albiventris (white-eared opossum). The results indicated that parasites with economic and public health relevance were present in wild animals, which may favor infection of humans and animals.(AU)

O uso de animais silvestres atropelados é uma estratégia eficiente para a pesquisa científica de patógenos. O objetivo deste estudo foi detectar DNA de parasitas do filo Apicomplexa em amostras de cérebro de animais silvestres atropelados nas mesorregiões Centro-Norte e Pioneira do Norte do Paraná, Brasil. Os transectos pré-estabelecidos foram percorridos semanalmente; quando encontrados, os animais foram armazenados em embalagens individuais e enviados para autópsia. Os fragmentos cerebrais foram coletados e mantidos a -20 ° C até o processamento. O DNA extraído das amostras foi amplificado por nested-PCR para o gene 18S rDNA do filo Apicomplexa. Todas as amostras positivas foram submetidas ao sequenciamento de DNA para definição da espécie. Um total de 90 animais foram coletados, no entanto, foram incluídos no estudo apenas 68 animais (75,6%) que apresentavam encéfalo. No sequenciamento foi possível identificar parasitos apicomplexos pelo sequenciamento de DNA em quatro amostras: Sarcocystis spp. em Colaptes melanochloros (pica-pau-verde-barrado) e em Mazama gouazoubira (veadocatingueiro); Neospora caninum em Leopardus pardalis (jaguatirica); e T. gondii em Didelphis albiventris (gambá-de-orelha-branca). Os resultados demonstraram que protozoários com relevância econômica e de saúde pública estavam presentes em animais silvestres, o que pode favorecer a infecção de humanos e animais.(AU)

Molecular detection of Apicomplexa protozoa in tissues from Alouatta guariba clamitans / Detecção molecular de protozoários Apicomplexa em tecidos de Alouatta guariba clamitans

The brown howler monkey (Alouatta guariba clamitans) is a primate species widely distributed in South America. Infections by protozoa are common in primates. However, studies on protozoa in primates in Brazil are scarce, so the goal of this study was to investigate DNA from the apicomplexan protozoa Neospora caninum, Sarcocystis spp. and Toxoplasma gondii in tissues of A. guariba clamitans. DNA extraction was performed on tissue samples from the heart, brain, liver, spleen, lung and intestine of six A. guariba clamitans from Santa Maria, Central Region of Rio Grande do Sul, Brazil. Conventional PCR was performed using 18S rRNA gene general primers for Apicomplexa and also specific primers to amplify Neosporaspp. and Toxoplasma gondii DNA. All animals were positive in the 18S PCR and the genetic sequencing confirmed the presence of Sarcocystis spp. DNA in the tissues of four animals belonging to at least two species (S. neurona and S. gigantea) and T. gondii DNA in the other two animals. One positive sample for T. gondii was genotypically characterized as atypical by the restriction fragment length polymorphism technique. N. caninum DNA was not detected in the tested samples. The presence of Apicomplexa protozoan DNA in the tissues of the six animals tested in this study highlights the importance of howler monkeys as maintainers of these pathogens in nature.(AU)

O bugio ruivo (Alouatta guariba clamitans) é uma espécie de primata amplamente distribuída na América do Sul. As infecções por protozoários são comuns em primatas. Entretanto, estudos sobre protozoários em primatas no Brasil são escassos, portanto o objetivo deste estudo foi pesquisar DNA dos protozoários Apicomplexa Neospora caninum, Sarcocystisspp. e Toxoplasma gondii em tecidos de A. guariba clamitans. A extração de DNA foi realizada em amostras de tecido do coração, cérebro, fígado, baço, pulmão e intestino de seis A. guariba clamitans oriundos de Santa Maria, Região Central do Rio Grande do Sul, Brasil. Foi realizada PCR convencional utilizando primers geral do gene 18S rRNA para Apicomplexa e também primers específicos para amplificação de DNA de Neospora spp.e Toxoplasma gondii. Todos os animais foram positivos no PCR geral para Apicomplexa e no sequenciamento genético confirmou-se a presença de DNA de Sarcocystis nos tecidos de quatro animais pertencentes a pelo menos duas espécies (S. neurona e S. gigantea), e DNA de T. gondii foi detectado nos outros dois animais. Uma amostra positiva para T. gondii foi caracterizada genotipicamente como atípico pela técnica de polimorfismo do comprimento do fragmento de restrição. Não foi detectado DNA de N. caninum nas amostras testadas. A presença de DNA de protozoários apicomplexa nos tecidos dos seis animais testados neste estudo destaca a importância dos bugios ruivos como mantenedores desses patógenos na natureza.(AU)

Molecular detection of Apicomplexa protozoa in tissues from Alouatta guariba clamitans / Detecção molecular de protozoários Apicomplexa em tecidos de Alouatta guariba clamitans

The brown howler monkey (Alouatta guariba clamitans) is a primate species widely distributed in South America. Infections by protozoa are common in primates. However, studies on protozoa in primates in Brazil are scarce, so the goal of this study was to investigate DNA from the apicomplexan protozoa Neospora caninum, Sarcocystis spp. and Toxoplasma gondii in tissues of A. guariba clamitans. DNA extraction was performed on tissue samples from the heart, brain, liver, spleen, lung and intestine of six A. guariba clamitans from Santa Maria, Central Region of Rio Grande do Sul, Brazil. Conventional PCR was performed using 18S rRNA gene general primers for Apicomplexa and also specific primers to amplify Neosporaspp. and Toxoplasma gondii DNA. All animals were positive in the 18S PCR and the genetic sequencing confirmed the presence of Sarcocystis spp. DNA in the tissues of four animals belonging to at least two species (S. neurona and S. gigantea) and T. gondii DNA in the other two animals. One positive sample for T. gondii was genotypically characterized as atypical by the restriction fragment length polymorphism technique. N. caninum DNA was not detected in the tested samples. The presence of Apicomplexa protozoan DNA in the tissues of the six animals tested in this study highlights the importance of howler monkeys as maintainers of these pathogens in nature.(AU)

O bugio ruivo (Alouatta guariba clamitans) é uma espécie de primata amplamente distribuída na América do Sul. As infecções por protozoários são comuns em primatas. Entretanto, estudos sobre protozoários em primatas no Brasil são escassos, portanto o objetivo deste estudo foi pesquisar DNA dos protozoários Apicomplexa Neospora caninum, Sarcocystisspp. e Toxoplasma gondii em tecidos de A. guariba clamitans. A extração de DNA foi realizada em amostras de tecido do coração, cérebro, fígado, baço, pulmão e intestino de seis A. guariba clamitans oriundos de Santa Maria, Região Central do Rio Grande do Sul, Brasil. Foi realizada PCR convencional utilizando primers geral do gene 18S rRNA para Apicomplexa e também primers específicos para amplificação de DNA de Neospora spp.e Toxoplasma gondii. Todos os animais foram positivos no PCR geral para Apicomplexa e no sequenciamento genético confirmou-se a presença de DNA de Sarcocystis nos tecidos de quatro animais pertencentes a pelo menos duas espécies (S. neurona e S. gigantea), e DNA de T. gondii foi detectado nos outros dois animais. Uma amostra positiva para T. gondii foi caracterizada genotipicamente como atípico pela técnica de polimorfismo do comprimento do fragmento de restrição. Não foi detectado DNA de N. caninum nas amostras testadas. A presença de DNA de protozoários apicomplexa nos tecidos dos seis animais testados neste estudo destaca a importância dos bugios ruivos como mantenedores desses patógenos na natureza.(AU)

High seroprevalence of Toxoplasma gondii and Neospora spp. in stray dogs from Rolim de Moura, Rondônia state, Western Brazilian Amazon / Alta soroprevalência de Toxoplasma gondii e Neospora spp. em cães errantes de Rolim de Moura, estado de Rondônia, Amazônia Ocidental Brasileira

Toxoplasma gondii and Neospora spp. are important apicomplexan pathogens that can infect dogs and result in a neurological syndrome. The aim of this study was to evaluate the seroprevalence of T. gondii and Neospora spp. in stray dogs in the state of Rondônia, Brazil. A cross-sectional study was conducted from June 2014 to April 2016. A total of 458 dogs blood samples were collected at the Center for Control of Zoonosis (CCZ), and anti-T. gondii and anti-N. spp. antibody levels were detected and measured using indirect fluorescent antibody test (IFAT). The seroprevalence of T. gondii and Neospora spp. in dogs it was 82.20% and 73.85%, respectively. The seroprevalence of coinfections was 47.59%. This is the first report of anti-T. gondii and anti-Neospora spp. antibodies detected in stray dogs in Rolim de Moura, state of Rondônia, Western Brazilian Amazon. Male dogs showed a higher frequency of anti-T. gondii antibodies than female dogs (88.0% vs. 75.3%; p < 0.001), presenting 2.41 times more chances of having the disease. High seroprevalence of infection (92.4%: T. gondii and 89.2%: Neospora spp.) was detected in the group ≥ 1-year-old dogs, in all collections, with OR 7.35 and OR 10.27 for the presence of anti-T. gondii and antiNeospora spp., respectively. In conclusion, the serological results indicate an important circulation of T. gondii and Neospora spp. in wandering dogs hailing from Rolim de Moura, state of Rondônia, Western Brazilian Amazonia.(AU)

Toxoplasma gondii e Neospora spp. são importantes patógenos apicomplexos que podem infectar cães e ocasionar síndrome neurológica. O objetivo deste estudo foi avaliar a soroprevalência de T. gondii e Neospora spp. em cães errantes do estado de Rondônia, Brasil. Um estudo transversal foi realizado durante o período de junho de 2014 a abril de 2016. Um total de 458 amostras de sangue de cães foram colhidas no Centro de Controle de Zoonoses (CCZ) e a detecção de anticorpos contra T. gondii e N. spp. foram realizados pela reação de imunofluorescência indireta (RIFI). A soroprevalência de T. gondii e Neospora spp. em cães foi 82,20% e 73,85%, respectivamente. A soroprevalência de coinfecções foi 47,59%. Este é o primeiro relato de anti-T. gondii e anti-Neospora spp. anticorpos detectados em cães errantes em Rolim de Moura, estado de Rondônia, Amazônia Ocidental Brasileira. Os cães machos apresentaram maior frequência de anticorpos anti-T. gondii do que as fêmeas (88,0% vs. 75,3%; p < 0,001), apresentando 2,41 vezes mais chance de ter a doença. Alta soroprevalência de infecção (92,4%: T. gondii e 89,2%: Neospora spp.) foi detectada no grupo ≥ cães com 1 ano de idade, em todas as coletas, com OR 7,35 e OR 10,27 para presença do anti-T. gondii e anti-Neospora spp., respectivamente. Em conclusão, os resultados sorológicos indicam uma importante circulação de T. gondii e Neospora spp. em cães errantes de Rolim de Moura, estado de Rondônia, Amazônia Ocidental Brasileira.(AU)

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.

Hepatozoon pyramidumi sp. n. (Apicomplexa: Adeleorina) from the blood of Echis pyramidum: morphology and SSU rDNA sequence / Hepatozoon pyramidumi sp. n. (Apicomplexa: Adeleorina) do sangue de Echis pyramidum: morfologia e sequência de SSU rDNA

Hepatozoon pyramidumi sp. n. is described from the blood of the Egyptian saw-scaled viper, Echis pyramidum, captured from Saudi Arabia. Five out of ten viper specimens examined (50%) were found infected with Hepatozoon pyramidumi sp. n. with parasitaemia level ranged from 20-30%. The infection was restricted only to the erythrocytes. Two morphologically different forms of intraerythrocytic stages were observed; small and mature gamonts. The small ganomt with average size of 10.7 × 3.5 μm. Mature gamont was sausage-shaped with recurved poles measuring 16.3 × 4.2 μm in average size. Infected erythrocytes were hypertrophied; their nuclei were deformed and sometimes displaced from their central position in the normal uninfected cell. Merogonic stages were observed in the lung endothelial cell and the liver parenchyma cells. Mature meront was 17.8 × 13.6 µm and contained banana-shaped merozoites with average size of ~15 × 2 µm. Phylogenetic analysis based on the SSU rDNA sequence clustered Hepatozoon pyramidumi sp. n with previously sequenced Hepatozoon spp., most of them infected reptilian hosts without geographic consideration. The morphological and molecular comparison with closely related species proved the taxonomic uniqueness and novelty of the present form.(AU)

Hepatozoon pyramidumi sp. n. é descrito a partir do sangue da víbora em escamas e quilhas serrilhadas, Echis pyramidum, capturada na Arábia Saudita. Cinco de dez espécimes de víbora examinadas (50%) foram encontradas infectadas com Hepatozoon pyramidumi sp. n. com nível de parasitemia de 20% a 30%. A infecção foi restrita apenas aos eritrócitos. Foram observadas duas formas morfologicamente diferentes de estágios intra-eritrocíticos: gamontes de tamanho pequeno e madura. As formas menores de gamontes apresentaram média de 10,7 × 3,5 μm. Os gamontes maduros apresentaram forma de salsicha, com pequenos polos recurvados, medindo 16,3 × 4,2 μm, em média. Os eritrócitos infectados estavam aumentados de tamanho; seus núcleos encontravam-se deformados e, algumas vezes, deslocados de sua posição central, quando comparados às células normais não-infectadas. Foram observados estágios merogônicos em células endoteliais pulmonares e nas células do parênquima hepático. Os merontes maduros apresentavam 17,8 × 13,6 µm e continham merozoítos em forma de banana com tamanho médio de ~ 15 × 2 µm. A análise filogenética baseada nas sequências SSU rDNA agrupou Hepatozoon pyramidumi sp. n com Hepatozoon spp. detectados em répteis de várias regiões geográficas. Por meio de análises morfológicas e moleculares com espécies intimamente relacionadas, demonstrou-se a singularidade dessa nova espécie de Hepatozoon.(AU)

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)

Occurrence of endo and hemoparasites in Sporophila caerulescens captured in the eastern region of the state of Acre, Brazil / Ocorrência de endo e hemoparasitos em Sporophila caerulescens capturados na região leste do estado do Acre, Brasil

Wild animals are hosts and reservoirs to many infectious agents, often unknown to the Scientific Community, which leads to serious health implications for natural and domestic environments. We conducted this research with the objective of diagnosing the occurrence of parasites of medical and veterinary interest in “coleirinhos” captured in the eastern region of the state of Acre, Brazil. We captured the specimens in August and September of 2017 using ornithological nets at the margins of Caeté river. We visually inspected the birds to evaluate the ectoparasites. Subsequently, we collected blood through a brachial vein puncture and rested the individuals in a cloth bag for 30 minutes to collect feces, after which we released them. Results of the visual inspection to detect and identify the ectoparasites were negative. Blood samples revealed the protozoan Haemoproteus sp., and the feces revealed cysts of Entamoeba histolytica and E. coli, oocysts of coccidia and eggs of Davaineidae. Coccidia was the most prevalent (69.2%) and abundant (34.15%) parasite reported. Parasites diagnosed in this study can promote the emergence of secondary infections in S. caerulescens individuals or other animals that are contaminated with these etiological agents, since, when migrating through different biomes of South America, other animal species can come into contact with these agents.(AU)

Os animais silvestres são hospedeiros e reservatórios de vários agentes infecciosos, muitas vezes desconhecidos para a comunidade científica, o que leva a sérias implicações na saúde dos ambientes naturais e domésticos. Esta pesquisa teve como objetivo diagnosticar a ocorrência de parasitos de interesse médico e veterinário em “coleirinhos” capturados no leste do Estado do Acre, Brasil. Os espécimes foram capturados nos meses de agosto e setembro de 2017 em redes ornitológicas as margens do Rio Caeté. Para a pesquisa de ectoparasitos, as aves foram inspecionadas visualmente. Após a inspeção, o sangue foi coletado por meio de punção da veia braquial e para coletas das fezes os indivíduos foram postos em descanso em saco de pano por 30min e depois foram soltos na natureza. As fezes, quando presentes, foram coletadas e acondicionadas em recipientes de plástico estéreis. A inspeção visual dos espécimes para detectar e identificar ectoparasitos resultou negativo. Nas amostras sanguíneas foi identificado o protozoário Haemoproteus sp.. Nas fezes foram encontrados os seguintes endoparasitos: cisto de Entamoeba histolytica e E. coli, oocisto de coccídeos e ovos de Davaineidae. Coccideos foram os endoparasitos mais prevalentes (69,2%) e os mais abundantes (34,15%). Os parasitos diagnosticados neste estudo podem propiciar o aparecimento de infecções secundárias nos indivíduos de S. caerulescens ou em outros animais que venham se contaminar com esses agentes etiológicos, pois com a migração dos mesmos, por diferentes biomas da América do Sul outras espécies de animais podem se infectar com esses agentes.(AU)

Toxoplasmose sistêmica em filhotes de gatos / Systemic toxoplasmosis in kittens

Background: Toxoplasmosis is a worldwide disease that affects virtually all species of warm-blooded animals. The felids,domestic and wild, are considered the definitive hosts of the protozoan. In Brazil, Toxoplasma gondii infection has beendiagnosed in horses, goats, primates, dogs and cats. In the backlands of Paraíba, the disease has been sporadically reportedaffecting dogs with canine distemper and swine, but cases of systemic toxoplasmosis in cats have not yet been described.The aim of the present study was to describe the main epidemiological, clinical and anatomopathological aspects of twocases of systemic toxoplasmosis in kittens.Cases: Two kittens were affected with three (cat 1) and six (cat 2) months old, females, and crossbreed. The kittens had notbeen vaccinated or dewormed, and were raised with other cats in a peridomiciliary regime in a rural area in the backlandsof Paraíba. Cat 1 was thin, apathetic, dehydrated, tachypneic and with pale mucous membranes. Cat 2, showed inappetence,apathy, jaundice, fever, dehydration, dyspnea and abdominal breathing pattern. At necropsy, non-collapsed, shiny, reddishlungs with multifocal whitish areas, punctuated or nodular, measuring from 0.1 to 0.3 cm in diameter, were found on thepleural surface and parenchyma. The livers were pale, with lobular pattern accentuation, and reddish depressed multifocalareas randomly distributed on the capsular surface. Histologically, multifocal to coalescent areas of necrosis, moderate (cat1) or marked (cat 2), associated with intralesional bradyzoites and tachyzoites and variable lymphoplasmacytic infiltratewere observed. In cat 2, numerous bradyzoites were visualized in the gray matter of the left cerebral hemisphere (temporaland parietal lobes), sometimes associated with a moderate lymphoplasmacytic inflammatory infiltrate. In the perivascularspaces of the...

Toxoplasmose sistêmica em filhotes de gatos / Systemic toxoplasmosis in kittens

Background: Toxoplasmosis is a worldwide disease that affects virtually all species of warm-blooded animals. The felids,domestic and wild, are considered the definitive hosts of the protozoan. In Brazil, Toxoplasma gondii infection has beendiagnosed in horses, goats, primates, dogs and cats. In the backlands of Paraíba, the disease has been sporadically reportedaffecting dogs with canine distemper and swine, but cases of systemic toxoplasmosis in cats have not yet been described.The aim of the present study was to describe the main epidemiological, clinical and anatomopathological aspects of twocases of systemic toxoplasmosis in kittens.Cases: Two kittens were affected with three (cat 1) and six (cat 2) months old, females, and crossbreed. The kittens had notbeen vaccinated or dewormed, and were raised with other cats in a peridomiciliary regime in a rural area in the backlandsof Paraíba. Cat 1 was thin, apathetic, dehydrated, tachypneic and with pale mucous membranes. Cat 2, showed inappetence,apathy, jaundice, fever, dehydration, dyspnea and abdominal breathing pattern. At necropsy, non-collapsed, shiny, reddishlungs with multifocal whitish areas, punctuated or nodular, measuring from 0.1 to 0.3 cm in diameter, were found on thepleural surface and parenchyma. The livers were pale, with lobular pattern accentuation, and reddish depressed multifocalareas randomly distributed on the capsular surface. Histologically, multifocal to coalescent areas of necrosis, moderate (cat1) or marked (cat 2), associated with intralesional bradyzoites and tachyzoites and variable lymphoplasmacytic infiltratewere observed. In cat 2, numerous bradyzoites were visualized in the gray matter of the left cerebral hemisphere (temporaland parietal lobes), sometimes associated with a moderate lymphoplasmacytic inflammatory infiltrate. In the perivascularspaces of the...(AU)

Rangelia vitalii infection in a dog from São Paulo city, Brazil: case report / Infecção por Rangelia vitalli em cão da zona norte de São Paulo/SP: relato de caso

Canine rangeliosis is an extravascular hemolytic disease caused by the protozoan Rangelia vitalii, which is transmitted by ticks of the species Amblyomma aureolatum. The most common clinical signs are apathy, hyperthermia and spontaneous bleeding. Anemia and thrombocytopenia are the most common hematological findings. This work reports a clinical case of canine Rangeliosis treated at a private veterinary hospital, in São Paulo city in 2017. A dog was treated at a veterinary hospital in the north of São Paulo, with progressive weight loss, apathy and tail injury. Anemia and thrombocytopenia were observed on the hemogram. Rangelia vitalii DNA was detected in animal blood by real-time PCR (qPCR). In addition to the supportive treatment, doxycycline and subcutaneous imidocarb applications were used. The sample collected after treatment with the antibiotic continued to present protozoal DNA. The disease should be considered as a differential diagnosis and there is a great need for further studies about the therapy used.(AU)

A rangeliose canina é uma doença hemolítica extravascular causada pelo protozoário Rangelia vitalii, o qual é transmitido por carrapatos da espécie Amblyomma aureolatum. Os sinais clínicos mais comuns são apatia, hipertermia e sangramentos espontâneos. Os achados hematológicos mais comuns são anemia e trombocitopenia. Este trabalho teve como objetivo relatar um caso clínico de Rangeliose canina tratada em um hospital veterinário particular, na cidade de São Paulo no ano de 2017. Um cão foi atendido em um hospital veterinário da zona norte de São Paulo, com emagrecimento progressivo, apatia e lesão na cauda. No hemograma foram observadas anemia e trombocitopenia. Através da PCR em tempo real (qPCR) do sangue do animal constatou-se a presença de DNA de Rangelia vitalii. Além do tratamento de suporte, utilizou-se doxiciclina e aplicações subcutâneas de imidocarb. A amostra coletada após o tratamento com o antibiótico continuou apresentando DNA do protozoário. A enfermidade deve ser considerada como diagnóstico diferencial e há uma grande necessidade de maiores estudos acerca da terapia utilizada.(AU)

Ultrastructure of phagocytes and oocysts of Nematopsis sp. (Apicomplexa, Porosporidae) infecting Crassostrea rhizophorae in Northeastern Brazil / Ultraestrutura de fagócitos e oocistos de Nematopsis sp. (Apicomplexa, Porosporidae) infectando Crassostrea rhizophorae no Nordeste brasileiro

This work describes the detailed ultrastructural morphology of the phagocyte imprisoning an oyster of Nematopsis(Apicomplexa) found in Crassostrea rhizophorae, in the city of Maceió (AL), Brazil. The highly infected hosts had half-open leaflets with weak, slow retraction of the adductor muscles. Variable number of ellipsoid oocytes, either isolated and or clustered, was found between myofibrils of the adductor muscle. Each oocyst was incarcerated in a parasitophorous vacuole of host uninucleated phagocyte. The oocysts were composed of a dense wall containing a uninucleate vermiform sporozoite. The wall of the fine oocysts was composed of homogeneous electron-lucent material formed by three layers of equal thickness, having a circular orifice-micropyle obstructed by the operculum. The oocysts presented ellipsoid morphology with their wall was surrounded by a complex network of numerous microfibrils. Important details of the taxonomic value were visualized such as the ultrastructural organization of the oocyst wall and the organization of the micropyle and operculum, beyond the microfibrils that protrude from the oocyst wall only observed by transmission electron microscopy (TEM) and that may aid in the identification of the species. However, in order to clarify the systematic position of the species reported of the genus Nematopsis, it is important to proceed with genetic analyses.(AU)

Este trabalho descreve a morfologia ultraestrutural detalhada do fagócito encarcerando um oocisto de Nematopsis(Apicomplexa) encontrado em Crassostrea rhizophorae, na cidade de Maceió (AL), Brasil. Os hospedeiros muito infectados apresentavam valvas entreabertas com retração fraca e lenta dos músculos abdutores. Número variável de oócitos de forma elipsoide, isolados e ou agrupados foi encontrado entre as miofibrilas do músculo abdutor. Cada oocisto estava encarcerado num vacúolo parasitóforo do fagócito uninucleado do hospedeiro. Os oocistos eram compostos por uma parede densa contendo um esporozoíto vermiforme uninucleado. A parede dos oocistos finos era composta de material electron-lucente homogêneo formado por três camadas de espessura igual, possuindo um orifício circular - micrópila, obstruída pelo opérculo. Os oocistos apresentavam morfologia elipsoide, sua parede era circundada por uma complexa rede de numerosas microfibrilas. Detalhes de valor taxonômico importantes foram visualizados tais como: a organização ultraestrutural da parede do oocisto e a organização da micrópila e do opérculo, além das microfibrilas que se projetam da parede do oocisto, estrutura apenas observada em microscopia eletrônica de transmissão (MET) e que pode auxiliar na identificação da espécie. Contudo, para esclarecer a posição sistemática da maioria das espécies relatadas do gênero Nematopsis é importante prosseguir com as análises genéticas.(AU)

Primeira descrição de pseudoparasitismo por esporocistos de Monocystis sp. em tatu-galinha, Dasypus novemcinctus (Linnaeus, 1758) - relato de caso / First description of pseudoparasitism by sporocysts of Monocystis sp. in nine banded armadillo, Dasypus novemcinctus (Linnaeus, 1758) - case report

O presente artigo é um relato sobre uma fêmea de D. novemcinctus que foi capturada de forma imprevista, durante um projeto de avaliação sanitária de animais silvestres sinantrópicos do município de Toledo/PR (autorização Sisbio/Ibama n. 55109-1). Durante a avaliação física, amostras de fezes foram coletadas após defecação espontânea e encaminhadas para análise. O material foi submetido aos métodos de flutuação simples em solução hipersaturada de cloreto de sódio e centrífugo-flutuação em sacarose. Na leitura das lâminas obtidas pelo método de centrífugo-flutuação em sacarose, foram observadas estruturas com comprimento de aproximadamente 10µm, forma bicônica e um tampão em cada extremidade, compatíveis com esporocistos de Monocystis sp. Protozoários do filo Apicomplexa, os Monocystis sp., são geralmente parasitas de anelídeos, como as minhocas. Esse organismo é comumente correlato a pseudoparasitismo em vários animais, porém não foram encontrados relatos de pseudoparasitismo desse agente em tatus-galinhas, logo essa é a primeira descrição de Monocystis sp. em espécime de D. novemcinctus.(AU)

The present article is an account of a female of D. novemcinctus which was captured during a sanitary evaluation project of synanthropic wild animals of the municipality of Toledo/PR (license SISBIO/ICMBIO n°. 55109-1). During a physical evaluation, samples of feces were collected, after spontaneous defecation, and sent for analysis. The material was subjected to simple flotation methods in hypersaturated sodium chloride solution and centrifugal-flotation solutions in sucrose. In a test reading by the centrifugal-flotation method in sucrose, structures with approximately 10µm, biconical shape and one plate at each end were observed, compatible with sporocysts of Monocystis sp. Protozoa of the phylum Apicomplexa, like Monocystis sp., it is common parasitic annelids, such as worms. This organism is a correlate of pseudoparasitism in several animals but was not found to describe this product as an agent in nine banded armadillos, so this is a first description of Monocystis sp. in specimen of D. novemcinctus.(AU)

Primeira descrição de pseudoparasitismo por esporocistos de Monocystis sp. em tatu-galinha, Dasypus novemcinctus (Linnaeus, 1758) - relato de caso / First description of pseudoparasitism by sporocysts of Monocystis sp. in nine banded armadillo, Dasypus novemcinctus (Linnaeus, 1758) - case report

O presente artigo é um relato sobre uma fêmea de D. novemcinctus que foi capturada de forma imprevista, durante um projeto de avaliação sanitária de animais silvestres sinantrópicos do município de Toledo/PR (autorização Sisbio/Ibama n. 55109-1). Durante a avaliação física, amostras de fezes foram coletadas após defecação espontânea e encaminhadas para análise. O material foi submetido aos métodos de flutuação simples em solução hipersaturada de cloreto de sódio e centrífugo-flutuação em sacarose. Na leitura das lâminas obtidas pelo método de centrífugo-flutuação em sacarose, foram observadas estruturas com comprimento de aproximadamente 10µm, forma bicônica e um tampão em cada extremidade, compatíveis com esporocistos de Monocystis sp. Protozoários do filo Apicomplexa, os Monocystis sp., são geralmente parasitas de anelídeos, como as minhocas. Esse organismo é comumente correlato a pseudoparasitismo em vários animais, porém não foram encontrados relatos de pseudoparasitismo desse agente em tatus-galinhas, logo essa é a primeira descrição de Monocystis sp. em espécime de D. novemcinctus.(AU)

The present article is an account of a female of D. novemcinctus which was captured during a sanitary evaluation project of synanthropic wild animals of the municipality of Toledo/PR (license SISBIO/ICMBIO n°. 55109-1). During a physical evaluation, samples of feces were collected, after spontaneous defecation, and sent for analysis. The material was subjected to simple flotation methods in hypersaturated sodium chloride solution and centrifugal-flotation solutions in sucrose. In a test reading by the centrifugal-flotation method in sucrose, structures with approximately 10µm, biconical shape and one plate at each end were observed, compatible with sporocysts of Monocystis sp. Protozoa of the phylum Apicomplexa, like Monocystis sp., it is common parasitic annelids, such as worms. This organism is a correlate of pseudoparasitism in several animals but was not found to describe this product as an agent in nine banded armadillos, so this is a first description of Monocystis sp. in specimen of D. novemcinctus.(AU)

Parasitism by Nematopsis sp. (Apicomplexa: Eugregarinida) in Mytella guyanensis at the Marine Extractive Reserve Baía do Iguape, Bahia, Brazil / Parasitismo por Nematopsis sp. (Apicomplexa: Eugregarinida) em Mytella guyanensis na Reserva Extrativista Marinha Baía do Iguape, Bahia, Brasil

Several studies show the presence of protozoa of the genus Nematopsis Schneider, 1892, in many species of bivalves. The pathogenicity of this Apicomplexa is still much debated, which is possibly related to the parasitism degree and host habitat. In this context, this study investigated parasitism by Nematopsis sp. in Mytella guyanensis (Bivalvia: Mytilidae) in a mangrove of the Marine Extractive Reserve Baía do Iguape, Bahia, Brazil. The collections were made monthly, from March 2014 to March 2015, consisting of 30 adults with shell height above 40 mm per month, totaling 360 specimens. The specimens were measured, weighed, opened, and macroscopically examined for identifying parasites and/or signs of morphological changes. After that, they were fixed in Davidson's solution and processed according to classical histological techniques, with inclusion in paraffin, obtaining 5 µm-thick sections by microtomy, and stained by Harris' Hematoxylin and Eosin (HE). The water temperature ranged from 25.5 to 33.6 ºC and the salinity from 21.1 to 34.3 Practical Salinity Units (PSU). The specimens measured between 41.1 and 68.6 mm and had an average weight of 7.24 g. Macroscopically, there was no presence of the parasite or any morphological changes related its presence. The frequency percentage of Nematopsis sp. was 99.45%. The mantle was the organ with the highest frequency of infection (46.26%), followed by the gonads (18.36%). Among the analyzed organs, all presented the parasite and, in a few severely infected cases, microscopical changes were observed in the mantle conformation. The infection intensity for most of the sessions analyzed was 1 to 3 oocysts/phagocyte; however, in one of the sessions, 9 oocysts/phagocytes were recorded. The parasite had a negative correlation with salinity. The high frequency of Nematopsis sp. should be considered for the sustainability of natural stocks, especially if such parasitism occurs simultaneously with other pathogens and stressful environmental conditions.(AU)

Diversos estudos evidenciam a presença de protozoários do gênero Nematopsis Schneider, 1892 em várias espécies de bivalves. A patogenicidade desse apicomplexo ainda é bastante debatida, a qual possivelmente relaciona-se ao grau de parasitismo e habitat do hospedeiro. Nesse contexto, este estudo investigou o parasitismo por Nematopsis sp. em Mytellaguyanensis(Bivalvia: Mytilidae) em um manguezal da Reserva Extrativista Marinha Baía do Iguape, Bahia, Brasil. As coletas foram efetuadas mensalmente, de março de 2014 a março de 2015, constando 30 adultos com altura da concha acima de 40 mm, totalizando 360 espécimens. Os espécimens foram medidos, pesados, abertos e examinados macroscopicamente para a identificação de parasitos e/ou sinais de alterações morfológicas, em seguida fixados em solução de Davidson e processados segundo técnica histológica clássica, com inclusão em parafina, obtenção de cortes de 5 µm de espessura por microtomia e coloração por hematoxilina de Harris e Eosina (HE). A temperatura da água variou de 25,5 a 33,6 ºC e a salinidade de 21,1 a 34,3 Unidades Práticas de Salinidade (UPS). Os espécimens mediram entre 41,1 e 68,6 mm e tiveram um peso médio de 7,24g. Macroscopicamente, não se observou presença do parasito ou qualquer alteração morfológica relacionada à presença do mesmo. A frequência percentual de Nematopsis sp. foi de 99,45%. O manto foi o órgão de maior frequência de infecção (46,26%), seguido das gônadas (18,36%). Entre os órgãos analisados, todos apresentaram o parasito, sendo que, para poucos casos severamente infectados, observaram-se microscopicamente modificações na conformação do manto. A intensidade de infecção para a maioria das sessões analisadas foi de 1 a 3 oocistos/fagócitos, contudo em uma das sessões foram registrados 9 oocistos/fagócitos. O parasito apresentou correlação negativa com a salinidade. A elevada frequência do Nematopsis sp. deve ser levada em consideração para a sustentabilidade dos estoques naturais, principalmente se tal parasitismo ocorrer simultaneamente a outros patógenos e condições ambientais estressantes.(AU)

Hepatozoon spp: relato de caso no oeste do Paraná-Brasil / Hepatozoon spp: case report in west Paraná-Brazil

O Hepatozoon spp. é um hemoparasita de cães, descrito em várias regiões do Brasil entre elas, São Paulo, Rio de Janeiro e Rio Grande do Sul, no entanto não há citações no estado do Paraná. Desta forma, relatou-se um caso de diagnóstico laboratorial de hepatozoonose trazendo informações sobre a doença e da importância do exame laboratorial pela visualização das inclusões em leucócitos pela microscopia óptica durante o hemograma.(AU)

Hepatozoon spp. is a hemoparasite of dogs, described in several regions of Brazil among them, São Paulo, Rio de Janeiro and Rio Grande do Sul, however there are no citations in the state of Paraná. In this way, a case of laboratory diagnosis of hepatozoonosis was reported, bringing information about the disease and the importance of the laboratory examination by the visualization of the inclusions in leukocytes by light microscopy during the hemogram.(AU)