Atypical Toxoplasma gondii genotype from a sheep and a pig on Fernando de Noronha Island, Brazil, showed different mouse virulence profiles.

Toxoplasma gondii is a zoonotic parasite which can infect almost all warm-blooded animals. Toxoplasma gondii isolates from Brazil have greater genetic diversity with a predominance of virulent and atypical genotypes, compared with the Northern Hemisphere. Considering that previous studies have demonstrated a high seroprevalence of T. gondii antibodies in animals from Fernando de Noronha Island, the aim of this study was to isolate, genetically characterize, and determine mouse virulence of isolates of T. gondii from livestock from this Brazilian island. Two T. gondii isolates were obtained by mouse bioassay from brain from one sheep and one pig. Genotyping was performed by PCR-RFLP using 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22- 8, c29-2, PK1, L358, and Apico) and an atypical genotype of T. gondii (ToxoDB #146) was identified for both isolates. Genotyping of four ROP loci indicated different alleles for ROP16 and mouse virulence analysis revealed different profiles (intermediate and low virulence). This is the first report of this genotype being described in a pig and a sheep.

Low frequency of antibodies anti-Neospora caninum in rodents in Fernando de Noronha Island, Brazil.

This study aimed to evaluate the frequency of Neospora caninum antibodies in rodents from the Island of Fernando de Noronha, Brazil, and to contribute to epidemiological data on neosporosis in this region. Blood samples were collected from 154 black-rats (Rattus rattus) and 12 rock-cavies (Kerodon rupestris). The search for anti-N. caninum antibodies was made using the NAT technique, with a cut-off of 1:20. The frequency of positive mice was 5.19% (8/154 - 1:20 titer) and all rock-cavies were negative. The low frequency of antibodies against N. caninum in rodents found in the Island shows the low contamination of the environment where these animals live.

Padronização de multiplex PCR para detecção de dermatófitos em pelos e crostas de cães e gatos / Standardization of a PCR multiplex for the detection of dermatophytes in dogs and cats fur and crusts

Objetivou-se padronizar uma reação do tipo multiplex PCR (mPCR) para detectar Microsporum canis, Microsporum gypseum e o complexo Trichophyton mentagrophytes em amostras de pelos e/ou crostas de cães e gatos. 250 amostras de pelos e/ou crostas de cães e gatos foram analisadas por meio de exame direto e cultura, o DNA das mesmas foi extraído para mPCR. Primers foram desenhados e como controle positivo da reação utilizou-se o DNA extraído de colônias de M. canis (URM 6273), M. gypseum (URM 6921) e T. mentagrophytes (URM 6211), provenientes da Coleção de Culturas (Micoteca URM), Departamento de Micologia, Centro de Ciências Biológicas da Universidade Federal de Pernambuco (CCB/UFPE). Como controles negativos de reação, utilizou-se água destilada esterilizada e DNA extraído de Alternaria sp. para verificar a especificidade dos primers. Do total de amostras analisadas, 15 (6%) foram identificadas, em cultura, como dermatófitos, e destas, 10 foram M. canis, três M. gypseum e dois T. mentagrophytes (complexo). Destas 15 amostras positivas, 11 (73,3%) foram detectadas por meio da mPCR. Além destas, seis outras, negativas em cultura, foram identificadas como M. gypseum. Verificou-se uma boa concordância entre os resultados da cultura e mPCR (Kappa: 0,66). O protocolo padronizado neste estudo pode ser utilizado como um método de triagem, por apresentar uma sensibilidade maior que a da cultura, usado paralelamente aos exames de rotina, permitindo um diagnóstico em menor tempo.(AU)

The aim of this study was to standardize a multiplex PCR (mPCR) reaction to detect Microsporum canis, Microsporum gypseum and the Trichophyton mentagrophytes complex in dog and cat fur and/or crusts. 250 fur and/or crusts samples from dogs and cats were analyzed by direct examination and culture, DNA from them was extracted for mPCR. Primers were designed and the DNA extracted from colonies of M. canis (URM 6273), M. gypseum (URM 6921) and T. mentagrophytes (URM 6211) from the Collection of Cultures - URM Micoteca - Department of Mycology, Biological Sciences Center of the Federal University of Pernambuco (CCB / UFPE). As negative controls, sterile distilled water and DNA extracted from Alternaria sp., were used to verify the specificity of the primers. Of the total samples analyzed, 15 (6%) were identified in culture as dermatophytes, and of these, 10 were M. canis, three M. gypseum and two T. mentagrophytes (complex). Of these 15 positive samples, 11 (73.3%) were detected by mPCR. Besides these, six others, negative in culture, were identified as M. gypseum. There was good agreement between culture results and mPCR (Kappa: 0.66). The protocol standardized in this study can be used as a screening method, because it has a sensitivity greater than that of the culture, used in parallel to the routine exams, allowing a diagnosis in a shorter time.(AU)

Distribuição espacial e abundância de carrapatos (Acari: Ixodidae) em remanescente de Mata Atlântica, Nordeste do Brasil / Spatial distribution and abundance of ticks (Acari: Ixodidae) in Atlantic forest remaining, northeastern Brazil

A distribuição espacial e a abundância de carrapatos em estágio de vida livre foram examinadas em fragmentos de Mata Atlântica no Nordeste do Brasil. O estudo foi realizado em Unidades de Conservação de Proteção Integral Mata do Tapacurá e Mata do Camucim, localizadas no município de São Lourenço da Mata, Pernambuco. Os carrapatos foram capturados pelo método da bandeira de flanela em seis parcelas de 200m2, sendo três parcelas estabelecidas em uma borda de mata adjacente a uma matriz agropastoril e outras três parcelas estabelecidas na borda de mata adjacente ao lago de barragem. Foram capturados 2652 carrapatos do gênero Amblyomma, sendo duas ninfas identificadas como Amblyomma dissimile com distribuição espacialmente agregada, demonstrados pela relação variância/média e o índice de agregação de Lloyd. Houve variação significativa na abundância de carrapatos entre as parcelas (H=26,79, p<0,005) sendo encontrados mais carrapatos na borda da mata adjacente à matriz agropastoril. Quanto as variáveis ambientais, houve correlação positiva entre a cobertura de dossel e a densidade de carrapatos (rs=0,31, p<0,05). No presente estudo observou-se que em estágios de vida livre, notadamente larvas e ninfas, a distribuição espacial e a abundância de carrapatos são fortemente influenciadas por áreas mais fechadas das matas. A maior presença de carrapatos próximos à matriz agropastoril poderia facilitar a migração de carrapatos entre ambientes silvestres e ambientes antrópicos.(AU)

The spatial distribution and abundance of free-living ticks were examined in Atlantic Forest fragments in northeastern Brazil. The study was conducted on Integral Protection Conservation Units of Mata do Tapacurá and Mata do Camucim, located in the municipality of São Lourenço da Mata, Pernambuco. Ticks were captured by flagging method in six plots of 200m2, three plots set in a forest edge adjacent to an agropastoral matrix and three plots set in the forest edge adjacent to a dam lake. Were captured 2652 ticks of the genus Amblyomma, two nymphs identified as Amblyomma dissimile with spatially aggregated distribution, shown by the variance/mean ratio and Lloyd aggregate index. There was significant variation in the abundance of ticks between plots (H=26.79, p<0.005) and more ticks was found in the woods adjacent to the edge agropastoral matrix. There was a positive correlation between canopy cover and tick density (rs=0.31, p<0.05). In the present study, it was observed that in free-living stages, especially larvae and nymphs, the spatial distribution and abundance of ticks are strongly influenced by more enclosed areas of forests. Most ticks near agropastoral matrix could facilitate the migration of ticks between wild environments and man-made or modified environments.(AU)

IDENTIFICATION OF BACTERIA IN BLOOD CULTURES FROM CLINICALLY ILL CAPTIVE ANTILLEAN MANATEES (TRICHECHUS MANATUS MANATUS).

Between September 2001 and March 2013, 62 bacterial cultures (37 aerobic and 25 anaerobic) were performed on 37 blood samples from 23 Antillean manatees ( Trichechus manatus manatus) that were kept in captivity at the Brazilian National Center for Research and Conservation of Aquatic Mammals (CMA) in Pernambuco (CMA-PE) and Alagoas (CMA-AL), Brazil. All of the animals sampled exhibited clinical signs at the time of sampling including abscesses (n = 8), debilitation and anorexia (n = 22), and profound lethargy-moribundity (n = 7). The 4 animals with profound lethargy-moribundity died shortly after sampling of unknown causes. Bacteria were isolated from 15/37 (40.5%) and aerobic blood cultures from 13/23 animals (56.5%). None of the anaerobic cultures were positive. Aeromonas caviae , Aeromonas hydrophila , Aeromonas sp., Escherichia coli , Leclercia adecarboxylata , Pantoea agglomerans , Pseudomonas aeruginosa , Pseudomonas stutzeri , Pseudomonas sp., Sphingomonas paucimobilis , coagulase-negative Staphylococcus, and Staphylococcus epidermidis were each found in only one animal; Staphylococcus spp. was found in two; and Vibrio fluvialis in four. Thirteen samples had only one bacteria isolated, one sample had two bacteria, and one sample had three bacteria isolated. Regarding sex, age group, and origin among the manatees examined, 54.5% (6/11) of the females, 58.3% (7/12) of the males, 40% (2/5) of the calves, 66.7% (8/12) of the juveniles, 50% (3/6) of the adults, 55.5% (10/18) at CMA-PE, and 60% (3/5) at CMA-AL were found to be positive for bacterial growth during at least one sampling time. All Antillean manatees were clinically ill. Regarding clinical signs, bacteria were found in 50% (11/22) of blood samples of the animals showing debilitation and anorexia, 1 of 8 (12.5%) of blood samples of the animals showing abscesses, and 3 of 7 (42.9%) of blood samples of the animals showing profound lethargy-moribundity.

Ectopic testis in coati (Nasua nasua Linnaeus, 1766) / Testículo ectópico em quati (Nasua nasua Linnaeus, 1766)

This paper reports a case of unilateral extracorporeal ectopic testes in a captive coati (Nasua nasua) in the State Park of Dois Irmãos Zoo, Recife/PE, Brazil. The testicle was located in the subcutaneous tissue of the inguinal region not adhered to the surrounding tissues. After bilateral orchiectomy, both testes were measured, fixed with 10% formalin buffered and embedded in paraffin for histopathological evaluation. The left testis measured 1.2 cm width by 1.7cm length, and the right one measured 1.5 cm width by 2.0 cm length. The ectopic testes had seminiferous epithelium without post-meiotic germ cell lines. The non-ectopic testis had several changes in the seminiferous epithelium that indicated degeneration. In both epididymis, the lumen did not contain sperm and the major epithelial structural alterations were more distinct in the epididymis associated to the ectopic testicle. In conclusion, the ectopic testis and epididymis had lesions compatible with testicular exposition to body temperature. Non-ectopic epididymis and testis had minor lesions but could be related to the infertility of the coati.(AU)

O artigo relata um caso de testículo ectópico em quati de cativeiro (Nasua nasua) no Zoológico do Parque Estadual Dois Irmãos, Recife/PE. O testículo encontrava-se localizado no tecido subcutâneo da região inguinal, sem estar aderido aos tecidos circunvizinhos. Após orquiectomia bilateral, ambos os testículos foram mensurados, fixados em formol a 10% e embebidos em parafina para avaliação histopatológica. O testículo esquerdo mediu 1,2cm de largura por 1,7cm de movimento; e o testículo direito mediu 1,5cm de largura por 2,0cm de comprimento. O testículo ectópico apresentou epitélio sem linhagem de células germinativas pós-meióticas. O testículo não ectópico apresentou alterações no epitélio seminífero caracterizando degeneração. Em ambos os epidídimos, o lúmen não continha espermatozoides e as principais alterações estruturais do epitélio foram mais distintas no epidídimo associado ao testículo ectópico. Conclui-se que o testículo ectópico e epidídimo apresentaram lesões características de aumento de temperatura. O testículo e epidídimo não ectópico apesentaram lesões menores mas que puderam ser associadas à infertilidade do quati.(AU)

Seroprevalence of Toxoplasma gondii among turkeys on family farms in the state of Northeastern Brazil.

The aim of the present study was to investigate the prevalence of antibodies against Toxoplasma gondii in turkeys and chickens on family farms in the semi-arid region of the state of Pernambuco, Brazil. In order to do so, 204 sera samples from turkeys were analyzed using the Modified Agglutination Test (MAT) and 322 sera samples from chickens were assessed using the Immunofluorescence Assay (IFA). Twenty-eight properties contained turkeys and 23 of these properties also contained chickens. The relative frequency of positive turkeys was 11% (21/204), with 46.6% (13/28) of the properties containing at least one positive turkey. The frequency of positive chickens was 25.8% (83/322), with 95.6% (22/23) of the properties containing at least one positive chicken. Based on the results of the present study, it was possible to conclude that turkeys can serve as an indicator of environmental contamination by oocysts of T. gondii. However, they are less effective than chickens bred in the same conditions. The increasing demand from consumers for naturally produced products should worry local sanitary authorities due to the high prevalence of antibodies against T. gondii found in this type of rearing system, particularly among chickens.