Molecular and serological detection of Leishmania spp. in captive wild animals from Ilha Solteira, SP, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mamíferos de médio e grande porte e sua relação com o mosaico de habitats na cuesta de Botucatu, Estado de São Paulo, Brasil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Molecular detection of Paracoccidioides brasiliensis in road-killed wild animals

Paracoccidioides brasiliensis infections have been little studied in wild and/or domestic animals, which may represent an important indicator of the presence of the pathogen in nature. Road-killed wild animals have been used for surveillance of vectors of zoonotic pathogens and may offer new opportunities for eco-epidemiological studies of paracoccidiodomycosis (PCM). The presence of P. brasiliensis infection was evaluated by Nested-PCR in tissue samples collected from 19 road-killed animals; 3 Cavia aperea (guinea pig), 5 Cerdocyon thous (crab-eating-fox), 1 Dasypus novemcinctus (nine-banded armadillo), 1 Dasypus septemcinctus (seven-banded armadillo), 2 Didelphis albiventris (white-eared opossum), 1 Eira barbara (tayra), 2 Gallictis vittata (grison), 2 Procyon cancrivorus (raccoon) and 2 Sphiggurus spinosus (porcupine). Specific P. brasiliensis amplicons were detected in (a) several organs of the two armadillos and one guinea pig, (b) the lung and liver of the porcupine, and (c) the lungs of raccoons and grisons. P. brasiliensis infection in wild animals from endemic areas might be more common than initially postulated. Molecular techniques can be used for detecting new hosts and mapping 'hot spot' areas of PCM.

Levantamento da mastofauna e identificação das influências antrópicas em dois fragmentos de Mata Atlântica transição com Cerrado de Rubião Junior – Botucatu - SP

Atlantic Forest and Cerrado are between the biological richest areas in the world, for that are considered top conservation priorities. Those ecosystems are found at Botucatu region, where it´s extremely fragmented by routes, agricultural areas and urban centres. This fragmentation causes restrictions on animals populations, in particular to medium and large mammals, that needs larger areas to live and also are often hunted. Even isolated and suffering anthropic influences the fragments are fundamental for local biodiversity conservation. We realized a mammal survey on two forest fragments of Atlantic forest transition to Cerrado of Rubião Júnior – Botucatu – SP – Brazil: Santo Antônio Church and Parque das Cascatas forests. The Santo Antônio church munt is considered a turistic point and its forest receive many visitants, Parque das Cascatas forest is inside of a residential condominium. Both areas are classify as semidecidual stational forest and are disconected by Domingos Sartori route, that connect Botucatu centre to Rubião Júnior district. Around both fragments can be found residences, plantations and pastures. The utilized method included: recognition of tracks on the forests, interviews with next residents and sand plots mounting. The interviews indicated 29 mammals species, and 19 was validated for animal traces, overruns or visualization. Other 3 species found were not mentioned on interviews. Totally, was found 22 small and medium mammals presence evidences on both forest fragments. Parque das Cascatas forest showed more diversity (19) then Santo Antônio Church forest (11), and species like Eira barbara, Procyon cancrivorus, Mazama americana, Shiggurus villosus, Dasypus novemcinctus, Didelphis albiventris, Lutreolina crassicaudata and Lepus europaeus looked for other areas once that was verifyed overruns of individuals... (Complete abstract click electronic access below)

Molecular detection of Leishmania spp. in road-killed wild mammals in the Central Western area of the State of São Paulo, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Susceptibility of Raccoons (Procyon lotor) to Infection with Mycobacterium bovis

Tuberculosis due to Mycobacterium bovis infection is endemic in white-tailed deer (Odocoileus virginianus) in the northeastern portion of the lower Michigan peninsula (USA). Various wild carnivores and omnivores, including raccoons (Procyon lotor), are infected with M. bovis within the endemic area. To investigate the pathogenesis of tuberculosis in raccoons and the likelihood of M. bovis transmission from infected raccoons to other susceptible hosts, we experimentally inoculated raccoons with single oral doses of M. bovis (ranging from 30 to 1.7 x 105 colony forming units [CFU]), five daily oral doses of M. bovis (ranging from 10 to 1 x 105 CFU), or a single intravenous (IV) dose of 1 x 105 CFU of M. bovis, from November 1998 through December 2000. Granulomatous lesions consistent with tuberculosis, or tissue colonization with M. bovis, were seen in one of five raccoons in the single low oral dose group, one of five raccoons in the multiple low oral dose group, two of five raccoons in the multiple medium oral dose group, five of five raccoons in the multiple high oral dose group, and five of five raccoons in the IV inoculated group. In orally inoculated raccoons, lesions were most common in the tracheobronchial and mesenteric lymph nodes and lung. Excretion of M. bovis in saliva or nasal secretions was noted in all IV inoculated raccoons and two of five multiple low oral dose raccoons. Mycobacterium bovis was not isolated from urine or feces from any experimentally inoculated raccoons. The need for multiple large oral doses to establish infection, and the low number of orally inoculated raccoons that excreted M. bovis in nasal secretions or saliva, suggest that widespread tuberculosis among raccoons is unlikely.

Landscape Genetics of Raccoons (Procyon lotor) Associated with Ridges and Valleys of Pennsylvania: Implications for Oral Rabies Vaccination Programs

Raccoons are the reservoir for the raccoon rabies virus variant in the United States. To combat this threat, oral rabies vaccination (ORV) programs are conducted in many eastern states. To aid in these efforts, the genetic structure of raccoons (Procyon lotor) was assessed in southwestern Pennsylvania to determine if select geographic features (i.e., ridges and valleys) serve as corridors or hindrances to raccoon gene flow (e.g., movement) and, therefore, rabies virus trafficking in this physiographic region. Raccoon DNA samples (n = 185) were collected from one ridge site and two adjacent valleys in southwestern Pennsylvania (Westmoreland, Cambria, Fayette, and Somerset counties). Raccoon genetic structure within and among these study sites was characterized at nine microsatellite loci. Results indicated that there was little population subdivision among any sites sampled. Furthermore, analyses using a model-based clustering approach indicated one essentially panmictic population was present among all the raccoons sampled over a reasonably broad geographic area (e.g., sites up to 36 km apart). However, a signature of isolation by distance was detected, suggesting that widths of ORV zones are critical for success. Combined, these data indicate that geographic features within this landscape influence raccoon gene flow only to a limited extent, suggesting that ridges of this physiographic system will not provide substantial long-term natural barriers to rabies virus trafficking. These results may be of value for future ORV efforts in Pennsylvania and other eastern states with similar landscapes.

Le Baylisascaris procyonis dans le sud du Québec : prévalence chez le raton laveur et étude de contamination des latrines

Le raton laveur (Procyon lotor) est porteur de nombreux parasites dont certains sont des agents de zoonose. Il nous est apparu opportun d’étudier la faune parasitaire intestinale du raton laveur québécois, particulièrement l’espèce Baylisascaris procyonis. L’étude a donc porté sur l’examen de matières fécales (n = 301) et de contenus intestinaux (n = 203) de 351 ratons laveurs provenant de deux programmes gouvernementaux sur la rage du raton laveur et d’excréments (n = 409) provenant de latrines dans des parcs nationaux du Québec. Des oeufs de B. procyonis étaient excrétés par 23,3 % des ratons laveurs, alors que 29,1 % étaient porteurs de vers. La charge parasitaire moyenne des vers B. procyonis de tout stade variait de 8,5 chez les adultes à 27,1 chez les juvéniles. Chez ces derniers, la charge parasitaire était 11,4 fois supérieure à celle des ratons adultes (p < 0,0001) dont 23,7 fois plus de vers immatures (p < 0,0001) et 9,1 fois plus de vers matures (p = 0,01). Le nombre d’oeufs était 24,7 fois plus élevé chez les ratons juvéniles (p = 0,02) et 50,0 fois moins élevé chez la femelle allaitante. La probabilité du raton d’excréter des oeufs l’automne était de 11,1 (IC95 %: 1,21-101,60) par rapport au printemps et de 21,1 (IC95 %: 2,91-153,18) par rapport à l’été (p < 0,01). La sensibilité (Se) et la spécificité (Sp) de la coproscopie comparée à la nécropsie (norme étalon) ont été calculées en se basant sur l’observation des vers matures (Se : 81,8 %; Sp : 97,7 %) et des vers de tout stade (Se : 53,9 %; Sp : 97,0 %). De plus, la recherche des parasites intestinaux nous a permis de découvrir des ookystes de Giardia (6/159), espèce parasitaire nouvellement rapportée chez le raton. En 2007, nous avons trouvé, par échantillonnage unique, des oeufs de B. procyonis dans 15,8 % des latrines (n = 165) identifiées dans des parcs nationaux du sud du Québec et en 2008, par échantillonnage multiple, dans 89,7 % des latrines (n = 26) actives situées dans des secteurs accessibles à l’homme dans trois parcs nationaux de la Montérégie. Le potentiel zoonotique de B. procyonis est un problème de santé publique qui pourrait devenir sérieux étant donné le pourcentage élevé de ratons laveurs qui excrètent le parasite et qui contaminent des secteurs accessibles par les humains.

Search for Mycobacterium leprae in wild mammals

Leprosy is still a worldwide public health problem. Brazil and India show the highest prevalence rates of the disease. Natural infection of armadillos Dasypus novemcinctus with Mycobacterium leprae has been reported in some regions of the United States. Identification of bacilli is difficult, particularly due to its inability to grow in vitro. The use of molecular tools represents a fast and sensitive alternative method for diagnosis of mycobacteriosis. In the present study, the diagnostic methods used were bacilloscopy, histopathology, microbiology, and PCR using specific primers for M. leprae repetitive sequences. PCR were performed using genomic DNA extracted from 138 samples of liver, spleen, lymph nodes, and skin of 44 D. novemcinctus, Euphractus sexcinctus, Cabassous unicinctus, and C. tatouay armadillos from the Middle Western region of the state of São Paulo and from the experimental station of Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Pantanal, located in Pantanal da Nhecolândia of Mato Grosso do Sul state. Also, the molecular analysis of 19 samples from internal organs of other road killed species of wild animals, such as Nasua nasua (ring-tailed coati), Procyon cancrivoros (hand-skinned), Cerdocyon thous (dog-pity-bush), Cavia aperea (restless cavy), Didelphis albiventris (skunk), Sphigurrus spinosus (hedgehog), and Gallictis vittata (ferret) showed PCR negative data. None of the 157 analyzed samples had shown natural mycobacterial infection. Only the armadillo inoculated with material collected from untreated multibacillary leprosy patient presented PCR positive and its genomic sequencing revealed 100% identity with M. leprae. According to these preliminary studies, based on the used methodology, it is possible to conclude that wild mammals seem not to play an important role in the epidemiology of leprosy in the Middle Western region of the São Paulo state and in the Pantanal of Mato Grosso do Sul state.

Fluoridation of the public water supply and prevalence of dental fluorosis in a peripheral district of the municipality of Bauru, SP

Objectives: The objectives of this study were to assess the fluoride concentration in the public water supply and the prevalence of dental fluorosis in schoolchildren between 7 and 15 years old, living in a peripheral district of the municipality of Bauru. Material and Methods: For this, fifty two water samples were collected on three different days of one week. These samples were analyzed for fluoride by means of the ion-sensitive electrode method (Orion 9609) coupled to a potentiometer (Procyon, model 720). In this method, 1.0 mL of TISAB II (Orion) was added to 1.0 mL of the sample. For the epidemiological survey of fluorosis, 52 schoolchildren of both genders, aged between 7 and 15 were assessed, with prior authorization from their caretakers. Only one person examined the children, after supervised toothbrushing and drying with cotton wool rolls. The TF index was used. Results: The fluoride concentrations in the water samples ranged from 0.62 to 1.20 mg/L, with a mean of 0.9 mg/L. The prevalence of dental fluorosis was 33%, with severity ranging from TF1 to TF4 (Kappa of 0.73 and concordance of 83.33%). Conclusions: The results from the analysis of water samples indicated a fluoride concentration greater than recommended for Bauru. The fluorosis levels found were higher than expected for a peripheral district, in which water is one of the few sources of fluoride.

Levantamento rápido de mamíferos terrestres de médio e grande porte no Parque Natural Municipal Cachoeira da Marta, Botucatu/SP

One of the most important strategies for biodiversity conservation is the establishment of protected areas, which protect areas important for the survival of several species. The park “Parque Natural Municipal Cachoeira da Marta”, located in Botucatu, São Paulo, is a unit of strictly protected area and aims to conserve the ecosystems and the biodiversity that exists in the area. The park has a management plan, but this does not include a survey of mammals present in the area, which is relevant as the group has a high degree of threat and a huge ecological importance. Thus, this study aimed to conduct a quick survey of terrestrial mammals of medium and large through the methodology of sand plots, with 25 plots distributed in five areas throughout the park. We recorded 4 orders, 6 families and 9 species (Nasua Nasua, Eira Barbara, Procyon cancrivorous, Leopardus pardalis, Leopardus sp., Dasypus novemcinctus, Tamandua tetradctyla, and Cavia aperea Myrmecophaga tridactyla). Most animals marked are generalist feeding habits, but it was also reported the presence of species more demanding regarding the quality of the habitat

Epizootiologic Survey of Mycobacterium Bovis in Wildlife and Farm Environments in Northern Michigan

Bovine tuberculosis (bovine TB), caused by Mycobacterium bovis, has reemerged in northern Michigan, USA, with detections in white-tailed deer (Odocoileus virginianus) in 1994 and in cattle in 1998. Since then, significant efforts have been directed toward reducing deer densities in the area in the hopes of reducing the bovine TB prevalence rate in deer and eliminating spillover of the disease into cattle. Despite the success of the efforts to reduce deer densities, additional cattle herds have become infected. Other mammals can be infected with M. bovis, and some carnivores and omnivores had been found to be infected with the disease in northern Michigan, USA. We conducted a multiyear surveillance effort to detect bovine TB in wild species of mammals in the Michigan, USA, outbreak area. From 2002 to 2004, tissue samples from 1,031 individual animals of 32 species were collected, processed, and cultured for M. bovis. Only 10 (1.0%) were culture-positive for M. bovis (five raccoons [Procyon lotor], four opossums [Didelphis virginiana], and one grey fox [Urocyon cinereoargenteus]). We also found two raccoons and four opossums to be positive for Mycobacterium avium. We collected 503 environmental samples from cattle farms recently identified as bovine TB positive; none yielded positive M. bovis culture results. Finally, we used infrared cameras to document wildlife use of four barns in the area. Many avian and mammalian species of wildlife were observed, with raccoons being the most commonly observed species. This surveillance study identified no new wildlife species that should be considered significant reservoirs of bovine TB in the outbreak area in northern Michigan, USA. However, the relatively high, apparent bovine TB prevalence rates in some carnivorous and omnivorous species, their relatively long life spans, and their frequent use of barns, suggests that removal of raccoons, opossums, foxes, and coyotes (Canis latrans) should be considered when a newly infected farm is depopulated of cattle.

Molecular epidemiology of animal rabies in the semiarid region of Paraíba, Northeastern Brazil

In the semiarid of the state of Paraíba, the anti-rabies vaccination is not common, most of the local inhabitants who deal with the animals do not know the incidence of the disease in the region. In this study, samples of foxes (Pseudalopex vetulus), insectivorous bats (Molossus molossus), raccoons (Procyon cancrivorous) and domestic animals brains were submitted to the diagnosis of rabies, by using the direct fluorescent antibody technique (d-FAT) and mouse inoculation test (MIT). Of the 581 examined materials, 50 (8.60 %) were positive for d-FAT and 47 (8.09 %) for MIT. From the positive samples for rabies, RNAs were extracted and transformed to cDNA, at the Laboratory of Rabies/Faculdade de Medicina Veterinária e Zootecnia/USP, SP. The phylogenetic characterization of the N gene was performed at the Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Medicina Veterinária Preventiva e Saúde Animal, Universidade Nihon, Faculdade de Ciências Bioresource, Fujisawa, Kanagawa, Japão. Based on the results of genotyping and phylogenetic analyzes, it is concluded that the epidemiology of rabies is complex in the semiarid of Paraíba, with different viral variants being maintained in domestic dogs, foxes, insectivorous bats and vampire bats. All the isolates examined belong to the genotype I of the genus Lyssavirus and it is possible to state that in the region, foxes are important sylvatic reservoirs of the rabies virus.

Distribución y taxonomía de mamíferos (clase mammalia) en el Cerro El Potosí Galeana, Nuevo León, México

El Cerro El Potosí es un área prioritaria para la Comisión Nacional para la Biodiversidad (CONABIO) y un Área Natural Protegida por el estado (Anónimo, 2000), debido a la existencia de una gran diversidad de tipos de vegetación y de especies de importancia fitogeografica o su orografía y altitud (3715 msnm) se considera un área promotora de procesos de especiación, registrándose especies endémicas y relictuales como el caso del Pinus culminicola y Nucifraga columbiana , es una de las montañas más altas del noroeste del país, está enclavada en la Sierra Madre Oriental. Con base en la información previa disponible, se presume que esta área ostente una mayor diversidad de flora y fauna. El objetivo principal fue el análisis de la diversidad y distribución de los mamíferos en los diferentes estratos vegetativos y altitudes dentro de los límites del Cerro El Potosí. Con el propósito de realizar las comparaciones de las diversidades de las especies en forma altitudinal, por vegetación, estacional; Se seleccionó la prueba de Chi-cuadrada para determinar dependencia entre especies y la altitud, tipo de muestreo o tipo de vegetación. Se efectuaron 24 salidas (mensuales) de campo, de noviembre del 2006 a octubre del 2008, de 3 a 4 días por salida. Se registraron 27 especies (14 Familias, 24 géneros, 27 especies, y 14 especies no reportadas previamente). De acuerdo al tipo de vegetación la riqueza se concentro en el bosque de pino seguido por el bosque de encino, las especies se distribuyeron en uno, en dos o más estratos de vegetación (patrones), para los diferentes tipos de vegetación se distribuyeron las especies de Silvilagus floridanus, Thomomys bottae, Peromyscus melanotis, Peromyscus levipes ambiguus, Canis latrans y Lynx rufus. Las especies que sólo se encontraron en un solo tipo de vegetación fueron Bassariscus astutus, Conepatus mesoleucus, Corynorhinus townsendii, Didelphis virginiana, Eptesicus fuscus, Puma yagouaroundi, Lasiurus cinereus, Lasiurus ega, Leptonycteris nivalis, Mustela frenata y Sorex milleri, por lo que se aprecia la diversidad encontrada en el sitio, esto en gran medida por la asociación de la vegetación y la altura marcado por la temperatura reportada para el sitio. Las especies que se localizaron en dos o más estratos o patrones de vegetación fueron Myotis thysanodes, Sciurus alleni, Otospermophilus variegatus, Microtus mexicanus, Urocyon cinereoargenteus, Procyon lotor, Puma concolor, Pecari tajacu y Odocoileus virginianus.