Anurans from the upper basins of the Rio Grande and Rio Preto: eastern portion of the Serra da Mantiqueira Environmental Protection Area, state of Minas Gerais, Brazil / Anfíbios anuros das bacias hidrográficas do alto rio Grande e alto rio Preto: porção leste da Área de Proteção Ambiental da Serra da Mantiqueira, estado de Minas Gerais, Brasil

The Neotropical zone stands out for having the greatest richness of amphibians in the world. About 625 species of anurans are known on Atlantic Forest. The southeastern mountainous regions of Brazil called Serra do Mar and Serra da Mantiqueira, are very important areas for the diversity of anurans in the country because they shelter remaining strata of the Atlantic Forest that have several endemic species. The present study aims to expand the knowledge about the Serra da Mantiqueira anurans, specifically from the Rio Preto and Rio Grande upper basins. The study area comprised 10 sample centers. We realized 38 campaigns, lasting two days each. During this period, we used active search as visual and auditory cues. The passive search was comprised of pitfall traps. Bioacoustic analyses were used to determine the cryptic species. The sufficiency of the sampling effort was estimated through the accumulation curve using Bootstrap and Jackknife-1 as estimators. During the study 55 species were found, 40 of them in the basin of the Rio Preto, and 44 in the basin of the Rio Grande. Twenty-nine species were found in both areas. The Hylidae family was predominant, followed by Leptodactylidae. In addition to species of the Atlantic Forest, the place houses typical components of the Cerrado biome and altitudinal swamps with unexpected sympatries. This inventory expands the area of occurrence of Scinax duartei, a species considered threatened. Although adjacent to the oldest national park in Brazil, the composition of anuran species of this area was unknown until then.

A região Neotropical se destaca por possuir a maior riqueza de anfíbios do mundo. Cerca de 625 espécies de anuros são conhecidas na Mata Atlântica. As regiões montanhosas do sudeste do Brasil, denominadas Serra do Mar e Serra da Mantiqueira, são áreas muito importantes para a diversidade de anuros no país, pois abrigam estratos remanescentes da Mata Atlântica que possuem diversas espécies endêmicas. O presente estudo visa ampliar o conhecimento sobre os anuros da Serra da Mantiqueira, especificamente das altas bacias dos rios Preto e Grande. A área de estudo compreendeu 10 centros amostrais. Realizamos 38 campanhas, com duração de dois dias cada. Nesse período, utilizamos a busca ativa como pistas visuais e auditivas. A busca passiva foi composta por armadilhas de queda. Análises bioacústicas foram usadas para determinar as espécies crípticas. A suficiência do esforço amostral foi estimada através da curva de acumulação usando Bootstrap e Jackknife-1 como estimadores. Durante o estudo foram encontradas 55 espécies, sendo 40 delas na bacia do Rio Preto e 44 na bacia do Rio Grande. Vinte e nove espécies foram encontradas em ambas as bacias. A família Hylidae foi predominante, seguida por Leptodactylidae. Além de espécies da Mata Atlântica, o local abriga componentes típicos do bioma Cerrado e brejos de altitude com simpatrias inesperadas. Este inventário amplia a área de ocorrência de Scinax duartei, espécie considerada ameaçada. Embora adjacente ao parque nacional mais antigo do Brasil, a composição de espécies de anuros dessa área era desconhecida até então.

Phallus eversion sexing in Phrynops geoffroanus (Testudines: Chelidae): a new non-invasive approach

Abstract The sexing of species is an important tool for population management. In tortoises and turtles, sexing is usually done according to secondary characteristics, such as plastron morphology or phallus eversion techniques. In this paper, we present the phallus eversion method and compare its efficiency to other existing techniques. This new method aims at shortening the management time and reducing the physical aftereffects on the animals, fostering the clinical and reproductive management of Geoffroys side-necked turtle (Phrynops geoffroanus) and related species. A sample of fifty individuals was taken in the central region of Piauí state, Northeastern Brazil. The proposed method consisted of moving the head from its resting position. The methods achieving a success rate higher than 80% were compared to each other as for the stimulation time required for phallus eversion. The method as here proposed achieved 100% efficiency in the sex determination of specimens with a linear carapace length between 10 and 34 cm. Also, when compared to the second most efficient method, it substantially reduced the time needed for managing the specimens (91%), avoiding possible sequelae in the individuals handled.

Resumo A sexagem das espécies é uma ferramenta importante para o manejo populacional. Em tartarugas, cágados e jabutis, a sexagem geralmente é feita de acordo com características secundárias, como morfologia do plastrão ou técnicas de eversão do falo. Neste artigo, apresentamos o método de eversão do falo e comparamos sua eficiência com outras técnicas existentes. Este novo método visa encurtar o tempo de manejo e reduzir os efeitos colaterais físicos nos animais, promovendo o manejo clínico e reprodutivo do cágado-de-barbicha (Phrynops geoffroanus) e espécies relacionadas. Uma amostra de cinquenta indivíduos foi coletada na região central do estado do Piauí, Nordeste do Brasil. O método proposto consiste em remover a cabeça de sua posição de repouso. Os métodos que obtiveram índice de sucesso superior a 80% foram comparados entre si quanto ao tempo de estimulação necessário para a eversão do falo. O método aqui proposto alcançou 100% de eficiência na determinação do sexo de espécimes com comprimento de carapaça linear entre 10 e 34 cm. Além disso, quando comparado ao segundo método mais eficiente, reduziu substancialmente o tempo necessário para o manejo dos espécimes (91%), evitando possíveis sequelas nos indivíduos manuseados.

Phallus eversion sexing in Phrynops geoffroanus (Testudines: Chelidae): a new non-invasive approach / Sexagem por eversão do falo em Phrynops geoffroanus (Testudines: Chelidae): uma nova abordagem não invasiva

Abstract The sexing of species is an important tool for population management. In tortoises and turtles, sexing is usually done according to secondary characteristics, such as plastron morphology or phallus eversion techniques. In this paper, we present the phallus eversion method and compare its efficiency to other existing techniques. This new method aims at shortening the management time and reducing the physical aftereffects on the animals, fostering the clinical and reproductive management of Geoffroy's side-necked turtle (Phrynops geoffroanus) and related species. A sample of fifty individuals was taken in the central region of Piauí state, Northeastern Brazil. The proposed method consisted of moving the head from its resting position. The methods achieving a success rate higher than 80% were compared to each other as for the stimulation time required for phallus eversion. The method as here proposed achieved 100% efficiency in the sex determination of specimens with a linear carapace length between 10 and 34 cm. Also, when compared to the second most efficient method, it substantially reduced the time needed for managing the specimens (91%), avoiding possible sequelae in the individuals handled.

Resumo A sexagem das espécies é uma ferramenta importante para o manejo populacional. Em tartarugas, cágados e jabutis, a sexagem geralmente é feita de acordo com características secundárias, como morfologia do plastrão ou técnicas de eversão do falo. Neste artigo, apresentamos o método de eversão do falo e comparamos sua eficiência com outras técnicas existentes. Este novo método visa encurtar o tempo de manejo e reduzir os efeitos colaterais físicos nos animais, promovendo o manejo clínico e reprodutivo do cágado-de-barbicha (Phrynops geoffroanus) e espécies relacionadas. Uma amostra de cinquenta indivíduos foi coletada na região central do estado do Piauí, Nordeste do Brasil. O método proposto consiste em remover a cabeça de sua posição de repouso. Os métodos que obtiveram índice de sucesso superior a 80% foram comparados entre si quanto ao tempo de estimulação necessário para a eversão do falo. O método aqui proposto alcançou 100% de eficiência na determinação do sexo de espécimes com comprimento de carapaça linear entre 10 e 34 cm. Além disso, quando comparado ao segundo método mais eficiente, reduziu substancialmente o tempo necessário para o manejo dos espécimes (91%), evitando possíveis sequelas nos indivíduos manuseados.

Anurans from the upper basins of the Rio Grande and Rio Preto: eastern portion of the Serra da Mantiqueira Environmental Protection Area, state of Minas Gerais, Brazil.

The Neotropical zone stands out for having the greatest richness of amphibians in the world. About 625 species of anurans are known on Atlantic Forest. The southeastern mountainous regions of Brazil called Serra do Mar and Serra da Mantiqueira, are very important areas for the diversity of anurans in the country because they shelter remaining strata of the Atlantic Forest that have several endemic species. The present study aims to expand the knowledge about the Serra da Mantiqueira anurans, specifically from the Rio Preto and Rio Grande upper basins. The study area comprised 10 sample centers. We realized 38 campaigns, lasting two days each. During this period, we used active search as visual and auditory cues. The passive search was comprised of pitfall traps. Bioacoustic analyses were used to determine the cryptic species. The sufficiency of the sampling effort was estimated through the accumulation curve using Bootstrap and Jackknife-1 as estimators. During the study 55 species were found, 40 of them in the basin of the Rio Preto, and 44 in the basin of the Rio Grande. Twenty-nine species were found in both areas. The Hylidae family was predominant, followed by Leptodactylidae. In addition to species of the Atlantic Forest, the place houses typical components of the Cerrado biome and altitudinal swamps with unexpected sympatries. This inventory expands the area of occurrence of Scinax duartei, a species considered threatened. Although adjacent to the oldest national park in Brazil, the composition of anuran species of this area was unknown until then.

Phallus eversion sexing in Phrynops geoffroanus (Testudines: Chelidae): a new non-invasive approach.

The sexing of species is an important tool for population management. In tortoises and turtles, sexing is usually done according to secondary characteristics, such as plastron morphology or phallus eversion techniques. In this paper, we present the phallus eversion method and compare its efficiency to other existing techniques. This new method aims at shortening the management time and reducing the physical aftereffects on the animals, fostering the clinical and reproductive management of Geoffroy's side-necked turtle (Phrynops geoffroanus) and related species. A sample of fifty individuals was taken in the central region of Piauí state, Northeastern Brazil. The proposed method consisted of moving the head from its resting position. The methods achieving a success rate higher than 80% were compared to each other as for the stimulation time required for phallus eversion. The method as here proposed achieved 100% efficiency in the sex determination of specimens with a linear carapace length between 10 and 34 cm. Also, when compared to the second most efficient method, it substantially reduced the time needed for managing the specimens (91%), avoiding possible sequelae in the individuals handled.

Efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e população de células natural killers na decídua e na glândula metrial de ratas / Effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats

Estudou-se o efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e na população de células natural killers (NK) na decídua e na glândula metrial de ratas durante a gestação. Avaliou-se a detecção imunoistoquímica de interferon γ (IFNγ), do fator inibidor de migração (MIF), da interleucina 15 (IL15), do óxido nítrico sintase induzível (iNOS), a marcação com lectina DBA para evidenciação das células NK uterinas DBA+ e a expressão gênica de Ifnγ e Nos2. O hipotireoidismo aumentou o iNOS aos sete dias, a IL15 e o MIF aos 10 e 12 dias, o IFNγ e o MIF aos 14 DG e a expressão dos transcritos gênicos para iNos aos 12 e 19 dias e para Ifnγ aos 14 DG. O hipotireoidismo reduziu a imunomarcação de MIF e lectina DBA aos sete dias, lectina DBA aos 10 e 14 DG, IFNγ aos 12 dias, e a expressão de Ifnγ aos 10 e 19 DG e de iNOS aos 12, 14 e 19 DG, bem como reduziu seus transcritos gênicos aos 10 e 14 DG. Conclui-se que o hipotireoidismo compromete o perfil imunológico na interface materno-fetal ao longo da gestação, particularmente por reduzir o fator anti-inflamatório iNOS e a população de células uNK DBA+.(AU)

The goal of this study was to evaluate the effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats. Interferon gamma (IFNγ), migration inhibitory factor (MIF), interleukin 15 (IL15), inducible nitric oxide synthase (iNOS), and DBA-Lectin labeling for evidence of uNK DBA+ cells in decidua and genetic expression of Ifnγ and iNos by real-time RT-PCR were evaluated. Hypothyroidism increased protein expression of iNOS at 7 days, IL15 and MIF at 10 and 12 days, IFNγ and MIF at 14 DG in the decidua and/or metrial gland and the gene transcripts for iNOS at 12 and 19 days and for Inf at 14 DG. In addition, hypothyroidism reduced the protein expression of MIF and DBA-Lectin at 7 days, DBA-Lectin at 10 and 14 DG, IFNγ at 12 days, and the gene transcript to Ifnγ at 10 and 19 DGs. Hypothyroidism also reduced the protein expression of iNOS at 12, 14 and 19 DG and reduced its gene transcripts at 10 and 14 DGs. It is concluded that hypothyroidism compromises the immunology profile at the maternal-fetal interface throughout pregnancy, particularly by reducing the anti-inflammatory factor iNOS and population of uNK DBA+ cells.(AU)

Efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e população de células natural killers na decídua e na glândula metrial de ratas / Effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats

Estudou-se o efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e na população de células natural killers (NK) na decídua e na glândula metrial de ratas durante a gestação. Avaliou-se a detecção imunoistoquímica de interferon γ (IFNγ), do fator inibidor de migração (MIF), da interleucina 15 (IL15), do óxido nítrico sintase induzível (iNOS), a marcação com lectina DBA para evidenciação das células NK uterinas DBA+ e a expressão gênica de Ifnγ e Nos2. O hipotireoidismo aumentou o iNOS aos sete dias, a IL15 e o MIF aos 10 e 12 dias, o IFNγ e o MIF aos 14 DG e a expressão dos transcritos gênicos para iNos aos 12 e 19 dias e para Ifnγ aos 14 DG. O hipotireoidismo reduziu a imunomarcação de MIF e lectina DBA aos sete dias, lectina DBA aos 10 e 14 DG, IFNγ aos 12 dias, e a expressão de Ifnγ aos 10 e 19 DG e de iNOS aos 12, 14 e 19 DG, bem como reduziu seus transcritos gênicos aos 10 e 14 DG. Conclui-se que o hipotireoidismo compromete o perfil imunológico na interface materno-fetal ao longo da gestação, particularmente por reduzir o fator anti-inflamatório iNOS e a população de células uNK DBA+.(AU)

The goal of this study was to evaluate the effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats. Interferon gamma (IFNγ), migration inhibitory factor (MIF), interleukin 15 (IL15), inducible nitric oxide synthase (iNOS), and DBA-Lectin labeling for evidence of uNK DBA+ cells in decidua and genetic expression of Ifnγ and iNos by real-time RT-PCR were evaluated. Hypothyroidism increased protein expression of iNOS at 7 days, IL15 and MIF at 10 and 12 days, IFNγ and MIF at 14 DG in the decidua and/or metrial gland and the gene transcripts for iNOS at 12 and 19 days and for Inf at 14 DG. In addition, hypothyroidism reduced the protein expression of MIF and DBA-Lectin at 7 days, DBA-Lectin at 10 and 14 DG, IFNγ at 12 days, and the gene transcript to Ifnγ at 10 and 19 DGs. Hypothyroidism also reduced the protein expression of iNOS at 12, 14 and 19 DG and reduced its gene transcripts at 10 and 14 DGs. It is concluded that hypothyroidism compromises the immunology profile at the maternal-fetal interface throughout pregnancy, particularly by reducing the anti-inflammatory factor iNOS and population of uNK DBA+ cells.(AU)

Estimation of a closed population size of tadpoles in temporary pond / Estimativa do tamanho da população fechada de girinos em lagoas temporárias

The practice of capture-recapture to estimate the diversity is well known to many animal groups, however this practice in the larval phase of anuran amphibians is incipient. We aimed at evaluating the Lincoln estimator, Venn diagram and Bayes theorem in the inference of population size of a larval phase anurocenose from lotic environment. The adherence of results was evaluated using the Kolmogorov-Smirnov test. The marking of tadpoles for later recapture and methods measurement was made with eosin methylene blue. When comparing the results of Lincoln-Petersen estimator corresponding to the Venn diagram and Bayes theorem, we detected percentage differences per sampling, i.e., the proportion of sampled anuran genera is kept among the three methods, although the values are numerically different. By submitting these results to the Kolmogorov-Smirnov test we have found no significant differences. Therefore, no matter the estimator, the measured value is adherent and estimates the total population. Together with the marking methodology, which did not change the behavior of tadpoles, the present study helps to fill the need of more studies on larval phase of amphibians in Brazil, especially in semi-arid northeast.(AU)

A prática de captura-recaptura para a estimação da diversidade é bem conhecida para diversos grupos animais, porém na fase larvar de anfíbios anuros essa prática é incipiente. Objetivamos avaliar os métodos do estimador de Lincoln, diagrama de Venn e o teorema de Bayes na inferência do tamanho populacional de uma anurocenose em fase larvar de ambiente lótico. A aderência dos resultados foi avaliada através do teste de Kolmogorov-Smirnov. A marcação dos girinos para posterior recaptura e aferição dos métodos foi feita com eosina de azul de metileno. Ao compararmos os resultados do estimador de Lincoln que corresponde com o do diagrama de Venn e com o teorema de Bayes detectamos diferenças percentuais por amostragem, isto é, a manutenção da proporção dos gêneros de anuros amostrados é mantida entre os três métodos, embora com valores numericamente diferentes. Ao submetermos esses resultados ao teste Kolmogorov-Smirnov não encontramos diferenças significativas. Logo, qualquer que seja o estimador o valor aferido é aderente e estima a população total. Aliado à metodologia de marcação que não alterou o comportamento dos girinos, o presente estudo ajuda a preencher a necessidade de mais estudos na fase larvar dos anfíbios no Brasil, em especial no semiárido nordestino.(AU)

Estimation of a closed population size of tadpoles in temporary pond / Estimativa do tamanho da população fechada de girinos em lagoas temporárias

Abstract The practice of capture-recapture to estimate the diversity is well known to many animal groups, however this practice in the larval phase of anuran amphibians is incipient. We aimed at evaluating the Lincoln estimator, Venn diagram and Bayes theorem in the inference of population size of a larval phase anurocenose from lotic environment. The adherence of results was evaluated using the Kolmogorov-Smirnov test. The marking of tadpoles for later recapture and methods measurement was made with eosin methylene blue. When comparing the results of Lincoln-Petersen estimator corresponding to the Venn diagram and Bayes theorem, we detected percentage differences per sampling, i.e., the proportion of sampled anuran genera is kept among the three methods, although the values are numerically different. By submitting these results to the Kolmogorov-Smirnov test we have found no significant differences. Therefore, no matter the estimator, the measured value is adherent and estimates the total population. Together with the marking methodology, which did not change the behavior of tadpoles, the present study helps to fill the need of more studies on larval phase of amphibians in Brazil, especially in semi-arid northeast.

Resumo A prática de captura-recaptura para a estimação da diversidade é bem conhecida para diversos grupos animais, porém na fase larvar de anfíbios anuros essa prática é incipiente. Objetivamos avaliar os métodos do estimador de Lincoln, diagrama de Venn e o teorema de Bayes na inferência do tamanho populacional de uma anurocenose em fase larvar de ambiente lótico. A aderência dos resultados foi avaliada através do teste de Kolmogorov-Smirnov. A marcação dos girinos para posterior recaptura e aferição dos métodos foi feita com eosina de azul de metileno. Ao compararmos os resultados do estimador de Lincoln que corresponde com o do diagrama de Venn e com o teorema de Bayes detectamos diferenças percentuais por amostragem, isto é, a manutenção da proporção dos gêneros de anuros amostrados é mantida entre os três métodos, embora com valores numericamente diferentes. Ao submetermos esses resultados ao teste Kolmogorov-Smirnov não encontramos diferenças significativas. Logo, qualquer que seja o estimador o valor aferido é aderente e estima a população total. Aliado à metodologia de marcação que não alterou o comportamento dos girinos, o presente estudo ajuda a preencher a necessidade de mais estudos na fase larvar dos anfíbios no Brasil, em especial no semiárido nordestino.

Estimation of a closed population size of tadpoles in temporary pond.

The practice of capture-recapture to estimate the diversity is well known to many animal groups, however this practice in the larval phase of anuran amphibians is incipient. We aimed at evaluating the Lincoln estimator, Venn diagram and Bayes theorem in the inference of population size of a larval phase anurocenose from lotic environment. The adherence of results was evaluated using the Kolmogorov-Smirnov test. The marking of tadpoles for later recapture and methods measurement was made with eosin methylene blue. When comparing the results of Lincoln-Petersen estimator corresponding to the Venn diagram and Bayes theorem, we detected percentage differences per sampling, i.e., the proportion of sampled anuran genera is kept among the three methods, although the values are numerically different. By submitting these results to the Kolmogorov-Smirnov test we have found no significant differences. Therefore, no matter the estimator, the measured value is adherent and estimates the total population. Together with the marking methodology, which did not change the behavior of tadpoles, the present study helps to fill the need of more studies on larval phase of amphibians in Brazil, especially in semi-arid northeast.

Validation of a reaction volume reduction protocol for analysis of Y chromosome haplotypes targeting DNA databases.

The use of Y chromosome haplotypes, important for the detection of sexual crimes in forensics, has gained prominence with the use of databases that incorporate these genetic profiles in their system. Here, we optimized and validated an amplification protocol for Y chromosome profile retrieval in reference samples using lesser materials than those in commercial kits. FTA® cards (Flinders Technology Associates) were used to support the oral cells of male individuals, which were amplified directly using the SwabSolution reagent (Promega). First, we optimized and validated the process to define the volume and cycling conditions. Three reference samples and nineteen 1.2 mm-diameter perforated discs were used per sample. Amplification of one or two discs (samples) with the PowerPlex® Y23 kit (Promega) was performed using 25, 26, and 27 thermal cycles. Twenty percent, 32%, and 100% reagent volumes, one disc, and 26 cycles were used for the control per sample. Thereafter, all samples (N = 270) were amplified using 27 cycles, one disc, and 32% reagents (optimized conditions). Data was analyzed using a study of equilibrium values between fluorophore colors. In the samples analyzed with 20% volume, an imbalance was observed in peak heights, both inside and in-between each dye. In samples amplified with 32% reagents, the values obtained for the intra-color and inter-color standard balance calculations for verification of the quality of the analyzed peaks were similar to those of samples amplified with 100% of the recommended volume. The quality of the profiles obtained with 32% reagents was suitable for insertion into databases.

Diversidade de carrapatos (Acari: Ixodidae) em animais silvestres recebidos pelo Zoológico Municipal de Guarulhos / Tick diversity (Acari: Ixodidae) on wild animals received by the Municipal Zoo of Guarulhos

O Zoológico Municipal de Guarulhos, localizado no estado de São Paulo, recebe e recupera animais silvestres do próprio município e de alguns municípios limítrofes. Entre os anos de 1997 e 2016 foram coletados carrapatos de animais silvestres (Cariama cristata, Buteo brachyurus, Alouatta guariba, Bradypus variegatus, Tamandua tetradactyla, Puma concolor, Cerdocyon thous, Nasua nasua, Mazama gouazoubira, Didelphis aurita, Sphiggurus villosus, Myocastor coypus e Hydrochoerus hydrochaeris) atendidos pelo Zoológico. Foram identificadas as seguintes espécies de carrapatos: Amblyomma aureolatum (Pallas, 1772), Amblyomma calcaratum Neumann, 1899, Amblyomma dubitatum Neumann, 1899, Amblyomma longirostre (Koch, 1844), Amblyomma nodosum Neumann, 1899, Amblyomma parkeri Fonseca e Aragão, 1952, Amblyomma sculptum Berlese, 1888, Amblyomma varium Koch, 1844, Haemaphysalis juxtakochi Cooley, 1946, Ixodes loricatus Neumann, 1899 e Rhipicephalus sanguineus sensu lato (Latreille, 1806). Apesar de todas as associações carrapato-hospedeiro já terem sido previamente relatadas no território brasileiro, o presente estudo registra pela primeira vez um ginandromorfo de A. dubitatum.(AU)

The Municipal Zoo of Guarulhos, located in the state of São Paulo, receives and recovers wild animals from the Guarulhos municipality and from some neighboring municipalities. Between the years of 1997 and 2016 ticks were collected on wild animals (Cariama cristata, Buteo brachyurus, Alouatta guariba, Bradypus variegatus, Tamandua tetradactyla, Puma concolor, Cerdocyon thous, Nasua nasua, Mazama gouazoubira, Didelphis aurita, Sphiggurus villosus, Myocastor coypus, and Hydrochoerus hydrochaeris) attended by the Zoo. The following tick species have been identified: Amblyomma aureolatum (Pallas, 1772), Amblyomma calcaratum Neumann, 1899, Amblyomma dubitatum Neumann, 1899, Amblyomma longirostre (Koch, 1844), Amblyomma nodosum Neumann, 1899, Amblyomma parkeri Fonseca and Aragão, 1952, Amblyomma sculptum Berlese, 1888, Amblyomma varium Koch, 1844, Haemaphysalis juxtakochi Cooley, 1946, Ixodes loricatus Neumann, 1899, and Rhipicephalus sanguineus sensu lato (Latreille, 1806). Although all tick-host associations have previously been reported in Brazilian territory, the present study records for the first time a gynandromorph of A. dubitatum.(AU)

DIVERSIDADE DE CARRAPATOS (ACARI: IXODIDAE) EM ANIMAIS SILVESTRES RECEBIDOS PELO ZOOLÓGICO MUNICIPAL DE GUARULHOS

O Zoológico Municipal de Guarulhos, localizado no estado de São Paulo, recebe e recupera animais silvestres do próprio município e de alguns municípios limítrofes. Entre os anos de 1997 e 2016 foram coletados carrapatos de animais silvestres (Cariama cristata, Buteo brachyurus, Alouatta guariba, Bradypus variegatus, Tamandua tetradactyla, Puma concolor, Cerdocyon thous, Nasua nasua, Mazama gouazoubira, Didelphis aurita, Sphiggurus villosus, Myocastor coypus e Hydrochoerus hydrochaeris) atendidos pelo Zoológico. Foram identificadas as seguintes espécies de carrapatos: Amblyomma aureolatum (Pallas, 1772), Amblyomma calcaratum Neumann, 1899, Amblyomma dubitatum Neumann, 1899, Amblyomma longirostre (Koch, 1844), Amblyomma nodosum Neumann, 1899, Amblyomma parkeri Fonseca e Aragão, 1952, Amblyomma sculptum Berlese, 1888, Amblyomma varium Koch, 1844, Haemaphysalis juxtakochi Cooley, 1946, Ixodes loricatus Neumann, 1899 e Rhipicephalus sanguineus sensu lato (Latreille, 1806). Apesar de todas as associações carrapato-hospedeiro já terem sido previamente relatadas no território brasileiro, o presente estudo registra pela primeira vez um ginandromorfo de A. dubitatum.

Diversidade de carrapatos (Acari: Ixodidae) em animais silvestres recebidos pelo Zoológico Municipal de Guarulhos / Tick diversity (Acari: Ixodidae) on wild animals received by the Municipal Zoo of Guarulhos

O Zoológico Municipal de Guarulhos, localizado no estado de São Paulo, recebe e recupera animais silvestres do próprio município e de alguns municípios limítrofes. Entre os anos de 1997 e 2016 foram coletados carrapatos de animais silvestres (Cariama cristata, Buteo brachyurus, Alouatta guariba, Bradypus variegatus, Tamandua tetradactyla, Puma concolor, Cerdocyon thous, Nasua nasua, Mazama gouazoubira, Didelphis aurita, Sphiggurus villosus, Myocastor coypus e Hydrochoerus hydrochaeris) atendidos pelo Zoológico. Foram identificadas as seguintes espécies de carrapatos: Amblyomma aureolatum (Pallas, 1772), Amblyomma calcaratum Neumann, 1899, Amblyomma dubitatum Neumann, 1899, Amblyomma longirostre (Koch, 1844), Amblyomma nodosum Neumann, 1899, Amblyomma parkeri Fonseca e Aragão, 1952, Amblyomma sculptum Berlese, 1888, Amblyomma varium Koch, 1844, Haemaphysalis juxtakochi Cooley, 1946, Ixodes loricatus Neumann, 1899 e Rhipicephalus sanguineus sensu lato (Latreille, 1806). Apesar de todas as associações carrapato-hospedeiro já terem sido previamente relatadas no território brasileiro, o presente estudo registra pela primeira vez um ginandromorfo de A. dubitatum.

The Municipal Zoo of Guarulhos, located in the state of São Paulo, receives and recovers wild animals from the Guarulhos municipality and from some neighboring municipalities. Between the years of 1997 and 2016 ticks were collected on wild animals (Cariama cristata, Buteo brachyurus, Alouatta guariba, Bradypus variegatus, Tamandua tetradactyla, Puma concolor, Cerdocyon thous, Nasua nasua, Mazama gouazoubira, Didelphis aurita, Sphiggurus villosus, Myocastor coypus, and Hydrochoerus hydrochaeris) attended by the Zoo. The following tick species have been identified: Amblyomma aureolatum (Pallas, 1772), Amblyomma calcaratum Neumann, 1899, Amblyomma dubitatum Neumann, 1899, Amblyomma longirostre (Koch, 1844), Amblyomma nodosum Neumann, 1899, Amblyomma parkeri Fonseca and Aragão, 1952, Amblyomma sculptum Berlese, 1888, Amblyomma varium Koch, 1844, Haemaphysalis juxtakochi Cooley, 1946, Ixodes loricatus Neumann, 1899, and Rhipicephalus sanguineus sensu lato (Latreille, 1806). Although all tick-host associations have previously been reported in Brazilian territory, the present study records for the first time a gynandromorph of A. dubitatum.

Human autosomal DNA and X chromosome STR profiles obtained from Chrysomya albiceps (Diptera: Calliphoridae) larvae used as a biological trace.

The use of insects to answer questions in criminal investigations, as well as a combination of forensic genetic techniques to obtain human DNA from the organisms, especially necrophagous dipterians, have gained ground in recent decades among researchers and professionals in this area. The objective of our study was to evaluate and compare two methods of human DNA extraction, commonly used for forensic samples, to obtain human autosomal DNA and X chromosome short tandem repeat profiles from the digestive tract of Chrysomya albiceps (Diptera: Calliphoridae) larvae. Immature specimens were collected from corpses at the Institute of Forensic Medicine of Pernambuco and raised in bovine ground meat to allow stabilization of the colony. Groups of larvae in the third instar were provided with bovine ground meat plus human blood for 48 h, dissected, and then subjected to DNA extraction. DNA was extracted using two methods: a DNA IQ™ kit and a phenol-chloroform method. Genomic DNA was amplified using AmpFℓSTR® Identifiler® Plus PCR and Argus-X-12® kits, and samples were sequenced to determine if the two extraction techniques generated reliable profiles that were compatible with a reference sample. The existence of comparable profiles from both techniques demonstrates the usefulness of dipteran larvae for obtaining human DNA from corpses, which can be further used to correlate genetic profiles in a crime scene when other traces are not available. However, several variables still require revision; thus, the technique should be further investigated for its validity, security, and, in particular, its reproducibility.

Detecting multiple DNA human profile from a mosquito blood meal.

Criminal traces commonly found at crime scenes may present mixtures from two or more individuals. The scene of the crime is important for the collection of various types of traces in order to find the perpetrator of the crime. Thus, we propose that hematophagous mosquitoes found at crime scenes can be used to perform genetic testing of human blood and aid in suspect investigation. The aim of the study was to obtain a single Aedes aegypti mosquito profile from a human DNA mixture containing genetic materials of four individuals. We also determined the effect of blood acquisition time by setting time intervals of 24, 48, and 72 h after the blood meal. STR loci and amelogenin were analyzed, and the results showed that human DNA profiles could be obtained from hematophagous mosquitos at 24 h following the blood meal. It is possible that hematophagous mosquitoes can be used as biological remains at the scene of the crime, and can be used to detect human DNA profiles of up to four individuals.