Prevalência e ciclo evolutivo de Isospora felis Wenyon, 1923 e I. rivolta (Grassi, 1879) Wenyon, 1923 em gatos

Foi levantada a prevalência de "I. felis" (24,0%), de "I. rivolta" (14,4%) e de infecções mistas (15,2%) em 125 gatos da Guanabara e do Rio de Janeiro, Brasil. Foi estudado o ciclo endógeno desses coccídios no epitélio intestinal. A "I. felis" se caracteriza pelo seu maior tamanho e pelo maior número de merozoítos e microgametos; e a "I. rivolta" por evoluir freqüentemente junto à lâmina própria, e quando isto não acontece, pelo seu menor tamanho em relação a "I. felis". Não foi conseguida a infecção de 3 cães desmamados e 4 recém-nascidos com as Isospora do gato. De 20 gatos com isosporose, apenas 2 deram reação de Sabin-Feldman positiva (1:16 e 1:64 respectivamente).

Prevalência da infecção pelo Toxoplasma gondii em animais domésticos, silvestres e grupamentos humanos da Amazônia

Através de Reação de Hemaglutinação Indireta para toxoplasmose foram examinadas amostras de sangue de dez diferentes espécies de animais domésticos e silvestres, de um grupamento humano da cidade de Manaus-Amazonas e de um grupamento humano indígena de área distante, no território de Roraima. Em 108 animais domésticos, o exame sorológico foi reagente em 90,6% dos gatos (Felis catus), 68,4% dos cães (Canis familiaris), 60,0% dos bovinos (Bos sp), 41,2% dos galináceos (Gallus sp) e 40,0% dos palmípedes (Cairina sp). Nos 104 animais silvestres foram reagentes 75,0% dos felídeos (Felis sp), 63,6% dos marsupiais (Didelphis marsupialis e Marmosa sp), 63,3% dos primatas (Saimiri sp) e 61,1% dos roedores (Proechimys). Entre os dois grupos humanos a prevalência foi de 70,6% nos 51 habitantes da área de Manaus, 64,8% nos 37 silvícolas de Roraima. Os autores discutem os resultados obtidos, assim como os diversos aspectos envolvidos na epidemiologia da toxoplasmose e chamam a atenção para a existência de mecanismos de transmissão ainda não esclarecidos, enfatizando a necessidade de maiores estudos dessa zoonose.

Wild Rodents as Experimental Intermediate Hosts of Lagochilascaris minor Leiper, 1909

A total of 25 specimens of Cavia porcellus (guinea pig), 5 Dasyprocta agouti (agouti), and 22 Calomys callosus (vesper mice) were inoculated with infective eggs of Lagochilascaris minor. The inoculum was prepared with embryonated eggs and orally administered to each individual animal through an esophagus probe. In parallel, 100 specimens of Felis catus domesticus were individually fed with 55-70 nodules containing 3rd-stage larvae encysted in tissues of infected rodents. Animals were examined and necropsied at different time intervals. The migration and encystment of L3 larva was observed in viscera, skeletal muscle, adipose and subcutaneous tissues from all rodents. Adult worms localized at abscesses in the cervical region, rhino, and oropharynx were recovered from domestic cats inoculated with infected rodent tissues. Through this study we can conclude that: (1) wild rodents act as intermediate hosts, characterizing this ascarid heteroxenic cycle; (2) in natural conditions rodents could possibly act as either intermediate hosts or paratenic hosts of Lagochilascaris minor; (3) despite the occurrence of an auto-infecting cycle, in prime-infection of felines (definite hosts) the cycle is only completed when intermediate hosts are provided; and (4) in the wild, rodents could serve as a source of infection for humans as they are frequently used as food in regions with the highest incidence of human lagochilascariasis.

Molecular identification of Rickettsia felis in ticks and fleas from an endemic area for Brazilian Spotted Fever

Rickettsioses are arthropod-borne diseases caused by parasites from the Order Rickettsiales. The most prevalent rickettsial disease in Brazil is Brazilian Spotted Fever (BSF). This work intends the molecular detection of those agents in ectoparasites from an endemic area of BSF in the state of Espírito Santo. A total of 502 ectoparasites, among them Amblyomma cajennense, Amblyomma dubitatum (A. cooperi), Riphicephalus sanguineus, Anocentor nitens and Ctenocephalides felis, was collected from domestic animals and the environment and separated in 152 lots according to the origin. Rickettsia sp. was detected in pools of all collected species by amplification of 17kDa protein-encoding gene fragments. The products of PCR amplification of three samples were sequenced, and Rickettsia felis was identified in R. sanguineus and C. felis. These results confirm the presence of Rickettsia felis in areas previously known as endemic for BSF, disease caused by Rickettsia rickettsii. Moreover, they show the needing of further studies for deeper knowledge of R. felis-spotted fever epidemiology and differentiation of these diseases in Brazil.

Antifungal activity of extracts from Atacama Desert fungi againstParacoccidioides brasiliensis and identification ofAspergillus felis as a promising source of natural bioactive compounds

Fungi of the genus Paracoccidioides are responsible for paracoccidioidomycosis. The occurrence of drug toxicity and relapse in this disease justify the development of new antifungal agents. Compounds extracted from fungal extract have showing antifungal activity. Extracts of 78 fungi isolated from rocks of the Atacama Desert were tested in a microdilution assay against Paracoccidioides brasiliensis Pb18. Approximately 18% (5) of the extracts showed minimum inhibitory concentration (MIC) values≤ 125.0 µg/mL. Among these, extract from the fungus UFMGCB 8030 demonstrated the best results, with an MIC of 15.6 µg/mL. This isolate was identified as Aspergillus felis (by macro and micromorphologies, and internal transcribed spacer, β-tubulin, and ribosomal polymerase II gene analyses) and was grown in five different culture media and extracted with various solvents to optimise its antifungal activity. Potato dextrose agar culture and dichloromethane extraction resulted in an MIC of 1.9 µg/mL against P. brasiliensis and did not show cytotoxicity at the concentrations tested in normal mammalian cell (Vero). This extract was subjected to bioassay-guided fractionation using analytical C18RP-high-performance liquid chromatography (HPLC) and an antifungal assay using P. brasiliensis. Analysis of the active fractions by HPLC-high resolution mass spectrometry allowed us to identify the antifungal agents present in the A. felis extracts cytochalasins. These results reveal the potential of A. felis as a producer of bioactive compounds with antifungal activity.

Role of Inflammatory Monocytes in Vaccine-Induced Reduction of Helicobacter felis Infection.

Despite the proven ability of immunization to reduce Helicobacter infection in mouse models, the precise mechanism of protection has remained elusive. In this study, we evaluated the role of inflammatory monocytes in the vaccine-induced reduction of Helicobacter felis infection. We first showed by using flow cytometric analysis that Ly6C(low) major histocompatibility complex class II-positive chemokine receptor type 2 (CCR2)-positive CD64(+) inflammatory monocytes accumulate in the stomach mucosa during the vaccine-induced reduction of H. felis infection. To determine whether inflammatory monocytes played a role in the protection, these cells were depleted with anti-CCR2 depleting antibodies. Indeed, depletion of inflammatory monocytes was associated with an impaired vaccine-induced reduction of H. felis infection on day 5 postinfection. To determine whether inflammatory monocytes had a direct or indirect role, we studied their antimicrobial activities. We observed that inflammatory monocytes produced tumor necrosis factor alpha and inducible nitric oxide synthase (iNOS), two major antimicrobial factors. Lastly, by using a Helicobacter in vitro killing assay, we showed that mouse inflammatory monocytes and activated human monocytes killed H. pylori in an iNOS-dependent manner. Collectively, these data show that inflammatory monocytes play a direct role in the immunization-induced reduction of H. felis infection from the gastric mucosa.

Variations of plasmid content in Rickettsia felis.

BACKGROUND: Since its first detection, characterization of R. felis has been a matter of debate, mostly due to the contamination of an initial R. felis culture by R. typhi. However, the first stable culture of R. felis allowed its precise phenotypic and genotypic characterization, and demonstrated that this species belonged to the spotted fever group rickettsiae. Later, its genome sequence revealed the presence of two forms of the same plasmid, physically confirmed by biological data. In a recent article, Gillespie et al. (PLoS One. 2007;2(3):e266.) used a bioinformatic approach to refute the presence of the second plasmid form, and proposed the creation of a specific phylogenetic group for R. felis. METHODOLOGY/PRINCIPAL FINDINGS: In the present report, we, and five independent international laboratories confirmed unambiguously by PCR the presence of two plasmid forms in R. felis strain URRWXCal(2) (T), but observed that the plasmid content of this species, from none to 2 plasmid forms, may depend on the culture passage history of the studied strain. We also demonstrated that R. felis does not cultivate in Vero cells at 37 degrees C but generates plaques at 30 degrees C. Finally, using a phylogenetic study based on 667 concatenated core genes, we demonstrated the position of R. felis within the spotted fever group. SIGNIFICANCE: We demonstrated that R. felis, which unambiguously belongs to the spotted fever group rickettsiae, may contain up to two plasmid forms but this plasmid content is unstable.

Aspectos morfométricos do timo em gatos domésticos (Felis domesticus)

O timo é um órgão linfático primário que desenvolve sua atividade em organismos jovens. Apesar de sua função ser responsável por mecanismos fundamentais na aquisição das defesas e conseqüentes respostas orgânicas, ela ainda não está totalmente esclarecida, nem tampouco as bases morfológicas que respondem por tais funções, como o processo de desenvolvimento e involução do órgão. Objetivou-se analisar e caracterizar os aspectos morfológicos do timo, tais como seu tamanho e volume, e aspectos histológicos do timo em gatos, correlacionando o sexo e o desenvolvimento etário. Doze timos provenientes de fetos de gatos domésticos (Felis domesticus) sem raça definida (SRD), machos e fêmeas, separados em três grupos etários. O timo apresentou-se com uma coloração rosa-pálida e com duas porções, a torácica e a cervical, sendo que cada uma delas possuía um lobo direito e um lobo esquerdo em sua maioria. A porção torácica localizava-se em região de mediastino cranial, entre os pulmões e à base do coração. E a porção cervical estendia-se além das costelas em sentido cranial, estando localizada ventralmente à traqueia. A estrutura celular do timo demonstrou-se organizada com a presença de agregados concêntricos, os chamados corpúsculos tímicos, formados por células epiteliais, sustentada por uma cápsula de tecido conjuntivo de onde partiam septos que ao penetrar no órgão dividia-o em lóbulos. Ocorreram variações significativas quanto à lobação e as dimensões do timo entre indivíduos da mesma faixa etária, e entre sexos diferentes. Os valores relativos ao comprimento, espessura e largura, de maneira geral, apresentaram aumento, em conformidade ao desenvolvimento dos animais, mas com diferenças entre os sexos.

Eficácia do fipronil em cães infestados com diferentes cargas parasitárias de Ctenocephalides felis felis (Siphonaptera: Pulicidae)

O objetivo do trabalho foi avaliar através de teste in vivo, a eficácia e o período residual de proteção do fipronil 10% "top spot" em cães infestados com diferentes cargas parasitárias de Ctenocephalides felis felis. Foram utilizados 24 cães da raça Beagle, compondo seis animais por grupo. Os cães foram divididos em quatro grupos. Os cães dos grupos controles I e II não receberam tratamento, enquanto que os cães dos grupos tratados I e II receberam tratamento com formulação de fipronil 10% "top spot". Os cães dos grupos controle I e tratado I foram infestados com 100 pulgas adultas não alimentadas, e os cães dos grupos controle II e tratado II foram infestados com 300 pulgas adultas não alimentadas. As infestações foram realizadas nos dias, -2, +5, +12, +19, +26, +33 e +40 e nos dias +2, +7, +14, +21, +28, +35 e +42 foi realizada retirada mecânica e contagem de pulgas para avaliação. As eficácias pulguicidas, para o grupo tratado I, nos dias +2, +7, +14, +21, +28, +35 e +42, foram respectivamente 99,36%; 99,73%; 99,48%; 99,74%; 99,75%; 95,06% e 67,62%. As eficácias pulguicidas, para o grupo tratado II, avaliadas nos mesmos dias, foram respectivamente 100%; 100%; 100%; 100%; 99,91%; 95,60% e 68,55%. O fipronil mostrou-se eficaz na eliminação das pulgas em cães até o dia +35. A análise estatística comparativa entre as médias de pulgas vivas, entre os grupos controle I e tratado I, demonstrou que ocorreu diferença significativa (p≤0,05) para os desafios em todos os dias experimentais, após o tratamento. Os grupos controle II e tratado II também apresentaram diferença significativa (p≤0,05) para os desafios em todos os dias experimentais, após o tratamento. A análise estatística entre os grupos tratados I e II demonstrou que não ocorreu diferença significativa (p≥0,05) para os desafios em todos os dias experimentais. O desafio foi encerrado no dia +42 já que a eficácia do fipronil nos grupos tratados I e II foram inferiores 70%. O produto em teste mostrou-se eficaz na eliminação das pulgas em cães até o dia + 35, não apresentando mais efeito residual de proteção quando os animais foram reinfestados. Não houve diferença significativa nos níveis de eficácia entre os grupos infestados com 100 e 300 exemplares adultos de C. felis felis ao longo do período experimental.

Predators of Spotted Flycatcher Muscicapa striata nests in southern England as determined by digital nest cameras

Capsule Avian predators are principally responsible. Aims To document the fate of Spotted Flycatcher nests and to identify the species responsible for nest predation. Methods During 2005-06, purpose-built, remote, digital nest-cameras were deployed at 65 out of 141 Spotted Flycatcher nests monitored in two study areas, one in south Devon and the second on the border of Bedfordshire and Cambridgeshire. Results Of the 141 nests monitored, 90 were successful (non-camera nests, 49 out of 76 successful, camera nests, 41 out of 65). Fate was determined for 63 of the 65 nests monitored by camera, with 20 predation events documented, all of which occurred during daylight hours. Avian predators carried out 17 of the 20 predations, with the principal nest predator identified as Eurasian Jay Garrulus glandarius. The only mammal recorded predating nests was the Domestic Cat Felis catus, the study therefore providing no evidence that Grey Squirrels Sciurus carolinensis are an important predator of Spotted Flycatcher nests. There was no evidence of differences in nest survival rates at nests with and without cameras. Nest remains following predation events gave little clue as to the identity of the predator species responsible. Conclusions Nest-cameras can be useful tools in the identification of nest predators, and may be deployed with no subsequent effect on nest survival. The majority of predation of Spotted Flycatcher nests in this study was by avian predators, principally the Jay. There was little evidence of predation by mammalian predators. Identification of specific nest predators enhances studies of breeding productivity and predation risk.

Preferência alimentar e identificação das principais fontes de repasto sanguíneo de fêmeas Lutzomyia (Diptera: Psychodidae) em áreas endêmicas para leishmaniose visceral na grande Natal

Leishmaniasis are endemic diseases wild spread in the New and Old World, caused by the flagelated protozoan Leishmania. In the New World, the distribution of different forms of leishmaniasis is mostly in tropical regions. In the State of Rio Grande do Norte, Northeast Brazil, 85% of the captured sand flies fauna is Lutzomyia longipalpis. The distribution of the sand fly vector in the state overlaps with the disease distribution, where the presence of sand flies is associated with presence of animals shelters. The aim of this study was to analyse the blood meal preference of sand flies vector from the genus Lutzomyia spp. in laboratory conditions, to verify the vector life cicle at different temperatures sets and to identify the main blood meal source in endemic areas for visceral leishmaniasis (VL) at peri-urban regions of Natal. Sand flies samples were collected from the municipalities of São Gonçalo do Amarante and Nísia Floresta where female sand flies were grouped for the colony maintenance in the laboratory and for the analysis of the preferred source of sand fly blood meal in natural environment. The prevalence of blood meal preference and oviposition for the females sand flies was 97% for Cavia porcellus with oviposition of 19 eggs/female; 97% for Eqqus caballus with 19 eggs/female; 98% for human blood with 14 eggs/female; 71.3% for Didelphis albiventris with 8.4 eggs/female; 73% for Gallus gallus with 14 eggs/female; 86% for Canis familiaris with 10.3 eggs/female; 81.4% for Galea spixii with 26 eggs/female; 36% for Callithrix jachus with 15 eggs/female; 42.8% for Monodelphis domestica with 0% of oviposition. Female sand flies did not take a blood meal from Felis catus. Sand flies life cycle ranged from 32-40 days, with 21-50 oviposition rates approximately. This study also showed that at 32°C the life cycle had 31 days, at 28° C it had 50 days and at 22°C it increased to 79 days. Adjusting the temperature to 35°C the eggs did not hatch, thus blocking the life cycle. A total of 1540 sand flies were captured, among them, 1.310 were male and 230 were female. Whereas 86% of the sand flies captured were Lu. longipalpis as compared to 10.5% for Lu. evandroi and, 3.2% for L. lenti and 0.3% for Lu whitmani. The ratio between female and male sandfly was approximately 6 males to 1 female. In Nísia Floresta, 50.7% of the collected females took their blood meal from armadillo, 12.8% from human. Among the female sand flies captured in São Gonçalo do Amarante, 80 of them were tested for the Leishmania KDNA infectivity where 5% of them were infected with Leishmania chagasi. Female Lutzomyia spp. showed to have an opportunistic blood meal characteristic. The behavioral parameters seem to have a higher influence in the oviposition when compared to the level of total proteins detected in the host s bloodstream. A higher Lu. longipalpis life cycle viability was observed at 28°C. The increase of temperature dropped the life cycle time, which means that the life cycle is modified by temperature range, source of blood meal and humidity. Lu longipalpis was the most specie found in the inner and peridomiciliar environment. In Nísia Floresta, armadillos were the main source of blood meal for Lutzomyia spp. At São Gonçalo do Amarante, humans were the main source of blood meal due to CDC nets placed inside their houses

Germ Cell Transplantation in Felids: A Potential Approach to Preserving Endangered Species

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cat flea (Ctenocephalides felis) infestation in quarantined marsh deer (Blastocerus dichotomus) populations

Marsh deer (Blastocenrs dichotomus) were captured for a research progam in Brazil and maintained in quarantine stations. After 60 days, fleas were detected on animals and identified as Ctenocephalides felis felis. Elimination of the infestation was difficult. Animal treatment with a fipronil-based compound was effective, and subsequently captured animals were treated prophylactically. Some animals remained infested, and some died from the infestation.