Hoary Fox Research Articles

Teste de ELISA indireto para diagnóstico sorológico de leishmaniose visceral em canídeos silvestres

Na América do Sul, alguns canídeos silvestres são considerados reservatórios naturais da Leishmania chagasi. A resposta imunológica desses animais à Leishmania é pouco conhecida, havendo a necessidade de métodos diagnósticos adequados para esse fim. No presente estudo, é descrita a padronização do ensaio imunoenzimático indireto (ELISA) para o diagnóstico sorológico de leishmaniose visceral em canídeos silvestres brasileiros. Foram estudadas amostras de soro e plasma de 12 canídeos cativos: sete lobos-guará (Chrysocyon brachyurus), três raposinhas (Lycalopex vetulus) e dois cachorros-do-mato (Cerdocyon thous). As amostras de um C. brachyurus e uma L. vetulus, cativos em área endêmica para LV, que apresentavam doença clínica e positividade em testes de Imunofluorescência Indireta e Reação em Cadeia de Polimerase, foram utilizadas como controles positivos. Foram comparados os conjugados anti-IgG de cão e proteína A, ambos ligados a peroxidase, cujos testes detectaram quatro (04/12) e três (03/12) C. brachyurus soropositivos para anticorpos anti-Leishmania sp., respectivamente. As médias das densidades ópticas (DOs) das amostras negativas foram nitidamente mais baixas do que as médias das DOs dos positivos tanto no ELISA com anti-IgG de cão (4,8 vezes) como com proteína A (15,5 vezes). Os soros de três C. brachyurus positivos no ELISA indireto foram avaliados por Western blotting e identificaram 22 bandas, sendo imunodominantes as de peso molecular de 19, 22, 24, 45 e 66 kDa. Os testes ELISA com a proteína A e o conjugado anti-IgG de cão apresentaram respectivamente concordância excelente (Kappa = 1; p<0,001) e moderada (Kappa = 0,8; p<0,0015), com o Western blotting. Ambos foram, portanto, considerados adequados a avaliações de triagem de animais cuja resposta humoral de anticorpos indica contato com o parasito, úteis para subsidiar estudos para adequação de metodologias específicas para os canídeos silvestres.

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.

SEROLOGIC SURVEY OF BRUCELLOSIS IN CAPTIVE NEOTROPICAL WILD CARNIVORES IN NORTHEAST BRAZIL

Abstract. This study reports the detection of antibodies against Brucella abortus and B. canis in wild neotropical carnivores kept in captivity in three zoos in northeastern Brazil. A total of 42 serum samples were examined, 17 from coatis (Nasua nasua), eight from crab-eating raccoons (Procyon cancrivorus), three from crab-eating foxes (Cerdocyon thous), three from hoary foxes (Lycalopex vetulus), two from little spotted cats (Leopardus tigrinus), five from tayras (Eira barbara), two from greater grisons (Galictis vittata), and two from neotropical river otters (Lontra longicaudis). The Rose-Bengal test and complement fixation test (CFT) were performed to detect anti-Brucella spp. antibodies, whereas the agar gel immunodiffusion test (AGID) was employed to detect anti-B. canis antibodies. The overall seroprevalence varied by species and by test; in addition, CFT and AGID seemed better able to detect antibodies against B. abortus and B. canis, respectively. This is the first study on the presence of anti-Brucella spp. antibodies in captive carnivores from Brazil, as well as the first report of antibodies to Brucella spp. in coatis, crab-eating raccoons, hoary foxes, little spotted cats, tayras, and greater grisons.

Heavy Metals in Hair of Wild Canids from the Brazilian Cerrado

In this study, we aimed to assess whether free-ranging wild canids are exposed to heavy metals in one of the most developed and populated regions of Brazil. Hair of 26 wild canids (maned wolves Chrysocyon brachyurus, crab-eating foxes Cerdocyon thous, and hoary foxes Lycalopex vetulus) from the Cerrado biome in Southeast Brazil were analyzed by spectrophotometry to detect cadmium, chromium, and lead, and also the essential copper, iron, manganese, and zinc traces. All samples showed traces of copper, iron, manganese, and zinc. Non-essential lead was detected in 57% (2.35 ± 0.99 mg/kg), and chromium in 88% (2.98 ± 1.56 mg/kg) of samples. Cadmium traces (detection limit 0.8 mg/kg) were not found. Crab-eating foxes had more copper, iron, and manganese in hair than maned wolves. Correlations among element levels differed between maned wolves and crab-eating foxes. Concentrations of chromium and lead were outstandingly higher than in wild canids from other areas. Addressing the causes of such levels and the impacts of the heavy metal pollution in Neotropical ecosystems is urgent for animal health and conservation purposes. We argue that heavy metal pollution should be considered as dangerous threats to wildlife health in Brazil and recommend hair sampling as a biomonitoring tool for heavy metals in Neotropical terrestrial mammals.

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

Leishmaniasis is a zoonotic disease that affects 12 million people worldwide. Several mammalian species can serve as a reservoir for this disease. Dogs are the main reservoir for visceral leishmaniasis in urban areas, which has become a serious public health concern in Brazil. The aim of this study was to evaluate the presence of Leishmania spp. in captive wild animals from Ilha Solteira, São Paulo, Brazil. Blood and various tissues samples were collected from animals of five different species: Speothos venaticus, Chrysocyon brachyurus, Cerdocyon thous, Pseudalopex vetulus, and Procyon cancrivorus. Antibodies against Leishmania spp. were detected in three wild canids by indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). PCR analyses of blood and bone marrow from all animals were negative, but Leishmania DNA was found in the tissues and skin of seropositive animals. Positive PCR samples were also positive for Leishmania donovani complex. Analysis of sequenced PCR products showed similarities with different regions of Leishmania (Leishmania) infantum and Leishmania (Leishmania) chagasi kinetoplastids. Measures to control visceral leishmaniasis in wild animals kept in Brazilian zoos should be established, as no disease control programs are currently available.

Ticks Infesting Wildlife Species in Northeastern Brazil With New Host and Locality Records

From September 2008 to March 2010, 397 ticks (315 larvae, 33 nymphs, 23 females, and 26 males) were collected from captive and free-living wildlife species in northeastern Brazil. Six tick species were identified, including Amblyomma auricularium (Conil) on Tamandua tetradactyla (L.), Amblyomma dubitatum Neumann on Hydrochaeris hydrochaeris (L.), Nectomys rattus (Pelzen) and T. tetradactyla, Amblyomma parvum Aragão on T. tetradactyla, Amblyomma rotundatum Koch on Boa constrictor L., Chelonoidis carbonaria (Spix), Kinosternon scorpioides (L.) and Rhinella jimi (Stevaux), Amblyomma oarium Koch on Bradypus variegatus Schinz, and Rhipicephalus sanguineus (Latreille) on Lycalopex vetulus (Lund). Nectomys rattus and T. tetradactyla are new hosts for A. dubitatum. This study extends the known distribution ofA. dubitatum in South America and provides evidence that its geographical range has been underestimated because of the lack of research. Four (A. dubitatum, A. parvum, A. rotundatum, and R. sanguineus) of six tick species identified in this study have previously been found on humans in South America, some of them being potentially involved in the transmission of pathogens of zoonotic concern.

Wild canids, domestic dogs and their pathogens in Southeast Brazil: disease threats for canid conservation.

Wild canids are under many pressures, including habitat loss, fragmentation and disease. The current lack of information on the status of wildlife health may hamper conservation efforts in Brazil. In this paper, we examined the prevalence of canine pathogens in 21 free-ranging wild canids, comprising 12 Cerdocyon thous (crab-eating fox), 7 Chrysocyon brachyurus (maned wolf), 2 Lycalopex vetulus (hoary fox), and 70 non-vaccinated domestic dogs from the Serra do Cipó National Park area, Southeast Brazil. For wild canids, seroprevalence of antibodies to canine parvovirus, canine adenovirus, canine coronavirus and Toxoplasma gondii was 100 (21/21), 33 (7/21), 5 (1/19) and 68 (13/19) percent, respectively. Antibodies against canine distemper virus, Neospora caninum or Babesia spp. were not found. We tested domestic dogs for antibodies to canine parvovirus, canine distemper virus and Babesia spp., and seroprevalences were 59 (41/70), 66 (46/70), and 42 (40/70) percent, respectively, with significantly higher prevalence in domestic dogs for CDV (P < 0.001) and Babesia spp. (P = 0.002), and in wild canids for CPV (P < 0.001). We report for the first time evidence of exposure to canine coronavirus in wild hoary foxes, and Platynossomun sp. infection in wild maned wolves. Maned wolves are more exposed to helminths than crab-eating foxes, with a higher prevalence of Trichuridae and Ancylostomidae in the area. The most common ectoparasites were Amblyomma cajennense, A. tigrinum, and Pulex irritans. Such data is useful information on infectious diseases of Brazilian wild canids, revealing pathogens as a threat to wild canids in the area. Control measures are discussed.

Composição e diversidade de mamíferos de médio e grande porte no Cerrado do Brasil central

Composição e diversidade de mamíferos de médio e grande porte no Cerrado do Brasil central. Estudos com mamíferos de médio e grande porte (&gt;1,0 kg) em áreas alteradas no Cerrado revelam que muitas espécies se adaptam aos ambientes antropizados. Foi investigada a estrutura da comunidade de mamíferos em uma área de 92.000 ha localizada no sudoeste baiano e constituída por plantios de Pinus spp., soja e faixas de cerrado sentido restrito. Os mamíferos foram registrados ao longo de nove campanhas de 12 dias, entre janeiro/2008 e maio/2009, no período diurno e noturno através de transectos lineares variando entre 5-15 km de extensão e que totalizaram 3.600 km percorridos/período. Foram registradas 29 espécies, sendo 23 através dos transectos lineares e as demais por observação indireta, captura e registros pretéritos. A comunidade é constituída em sua maioria por representantes da ordem Carnivora (41,4%) e espécies onívoras (36,7%), sendo Lycalopex vetulus a única espécie endêmica do Cerrado registrada na área. Apenas 13,6% das espécies foram frequentes em todas as campanhas (Ozotoceros bezoarticus, Pecari tajacu e Tolypeutes tricinctus) e sete delas correspondem a 91,73% dos registros realizados. O tatu-bola (T. tricinctus) representou o maior número de registros (23,76%) e provavelmente a área de estudo abriga a maior população dessa espécie ameaçada no bioma, destacando a importância em evitar que as áreas de vegetação nativa sejam substituídas por monoculturas e reflorestamento. Muitas das espécies registradas são generalistas no uso do habitat e amplamente distribuídas pelo bioma, ocorrendo inclusive em ambientes alterados. Considera-se que a manutenção da diversidade observada na área é decorrente de sua grande extensão e do mosaico de ambientes que proporcionam uma maior variedade e diversidade de recursos, como abrigo e alimento, possibilitando inclusive a presença de espécies com baixas densidades e especialistas na dieta e habitat.

First Identification of Canine Distemper Virus in Hoary Fox (Lycalopex vetulus): Pathologic Aspects and Virus Phylogeny

Canine distemper virus (CDV) has been reported in several wild animal species, but there have been no reports of CDV in hoary fox (Lycalopex vetulus). This paper characterizes the first case of natural CDV infection in hoary fox, including the clinical and pathologic aspects of the disease as well as the viral strain phylogeny.

DNA extraction from hair shafts of wild Brazilian felids and canids

Wild felids and canids are usually the main predators in the food chains where they dwell and are almost invisible to behavior and ecology researchers. Due to their grooming behavior, they tend to swallow shed hair, which shows up in the feces. DNA found in hair shafts can be used in molecular studies that can unravel, for instance, genetic variability, reproductive mode and family structure, and in some species, it is even possible to estimate migration and dispersion rates in given populations. First, however, DNA must be extracted from hair. We extracted successfully and dependably hair shaft DNA from eight wild Brazilian felids, ocelot, margay, oncilla, Geoffroy's cat, pampas cat, jaguarundi, puma, and jaguar, as well as the domestic cat and from three wild Brazilian canids, maned wolf, crab-eating fox, and hoary fox, as well as the domestic dog. Hair samples came mostly from feces collected at the São Paulo Zoo and were also gathered from non-sedated pet or from recently dead wild animals and were also collected from museum specimens. Fractions of hair samples were stained before DNA extraction, while most samples were not. Our extraction protocol is based on a feather DNA extraction technique, based in the phenol:chloroform:isoamyl alcohol general method, with proteinase K as digestive enzyme.

Lycalopex vetulus (Carnivora: Canidae)

Lycalopex vetulus (Lund, 1842), commonly called the hoary fox, is Brazil's smallest canid. It has a slender body and limbs and a small skull composed of relative fragile bones. Its size and skull characteristics combined with its peculiar dentition make it more suitable for an insectivorous diet rather than one of large vertebrates. L. vetulus is endemic to Brazil and occurs primarily in the south-central cerrados at low elevations. There is some evidence to indicate its presence in Bolivia. L. vetulus is widespread and abundant in Brazil and presently there are no known threats that would result in declines in its occurrence in the core area of its distribution.

Anticorpos anti Brucella abortus, anti Brucella canis e anti Leptospira spp. em raposas (Pseudalopex vetulus) do semiárido paraibano, Nordeste do Brasil

O objetivo do presente trabalho foi determinar a ocorrência de anticorpos anti Brucella abortus, anti Brucella canis e anti Leptospira spp. em raposas (Pseudalopex vetulus). Para tanto, foram utilizadas 60 raposas atropeladas em rodovias no semiárido da Paraíba, Nordeste do Brasil. Para a detecção de anticorpos anti Brucella abortus, o teste do antígeno acidificado tamponado (AAT) foi empregado como teste de triagem, e a prova do 2-mercaptoetanol foi empregada como método confirmatório. Para o diagnóstico sorológico das infecções por Brucella canis e Leptospira spp., foram utilizados os testes de imunodifusão em gel de agar (IDGA) e soroaglutinação microscópica, respectivamente. Todas as amostras foram negativas na pesquisa de anticorpos anti Brucella canis e anti Leptospira spp. Das 60 raposas testadas, 16 (26,6%) foram positivas para anticorpos anti Brucella abortus no teste de AAT, e quatro (6,7%) amostras foram confirmadas no teste de 2-mercaptoetanol, sendo duas amostras com título 100 e duas com título 50.

Complete genome analysis of a rabies virus isolate from Brazilian wild fox

The complete genome sequence of wild-type rabies virus (RABV) isolated from a wild Brazilian hoary fox (Dusicyon sp.), the BR-Pfx1 isolate, was determined and compared with fixed RABV strains. The genome structure and organization of the BR-Pfx1 isolate were composed of 11,924 nt and included the five standard genes of rhabdoviruses. Sequences of mRNA start and stop signals for transcription were highly conserved among all structural protein genes of the BR-Pfx1 isolate. All amino acid residues in the glycoprotein (G) gene associated with pathogenicity were retained in the BR-Pfx1 isolate, while unique amino acid substitutions were found in antigenic region I of the nucleoprotein gene and III of G. These results suggest that although the standard genome structure and organization of the RABV isolate are common between the BR-Pfx1 isolate and fixed RABV strains, the unique amino acid substitutions in functional sites of the BR-Pfx1 isolate may result in different biological characteristics from fixed RABV strains.

Serological survey for Leishmania sp. infection in wild animals from the municipality of Maringá, Paraná state, Brazil

Leishmania sp. infection was investigated in wild animals from the Ingá Park, in the municipality of Maringá, Paraná state, Brazil, where American cutaneous leishmaniasis (ACL) is an endemic disease. Sixty-five mammals, comprising Didelphis albiventris, Cerdocyon thous, Lycalopex vetulus, Cebus apella, Dasyprocta azarae, Dasypus novemcinctus, Procyon cancrivorus and Nasua nasua, were captured. Blood samples were collected for parasite cultivation. Antibodies were investigated by direct agglutination test (DAT) using Leishmania (Viannia) braziliensis as antigen. Flagellates were observed in blood cultures of 14 (35.9%) Didelphis albiventris. Anti-Leishmania antibodies were detected in 31 (51.6%) specimens of Cerdocyon thous, Lycalopex vetulus, Cebus apella, Dasyprocta azarae, Procyon cancrivorus and Nasua nasua. These results suggest that Cerdocyon thous and Lycalopex vetulus (crab-eating fox), Cebus apella (capuchin monkey), Dasyprocta azarae (agouti), Procyon cancrivorus (crab-eating raccoon) and Nasua nasua (coati) play an important role in the ACL transmission cycle in the northwestern region of Paraná, Brazil.

Isolation of rabies virus from the parotid salivary glands of foxes (Pseudalopex vetulus) from Paraíba State, Northeastern Brazil

To determine the presence of rabies virus in the parotid salivary glands, 12 road-killed rabies-positive hoary foxes (Pseudoalopex vetulus) were tested by using the fluorescent antibody test (FAT) and mouse inoculation test (MIT). All 12 parotid salivary glands were positive for both tests, although in some cases several passages were required. The findings of this study support the importance of the hoary fox as rabies reservoir in the sem-iarid region of Paraíba State, Northeastern Brazil.

Visceral leishmaniasis in captive wild canids in Brazil

Visceral leishmaniasis (VL) is endemic in Belo Horizonte (State of Minas Gerais, Brazil). Leishmania sp. can naturally infect several species of mammals, and the domestic dog is the most important reservoir of the disease in South America. This report describes five cases of visceral leishmaniasis in Brazilian canids. Among 15 animals kept in captivity in a zoo in Belo Horizonte (State of Minas Gerais, Brazil), two animals, a bush dog ( Spheotos venaticos) and a hoary zorro ( Lycalopex vetulus) were serologically positive and developed clinical signs of VL, whereas three other canids, including a crab-eating fox ( Cerdocyon thous), a maned wolf ( Chrysocyon brachyurus), and a hoary zorro ( Lycalopex vetulus) had positive serological results without clinical signs.

Densidade populacional de raposa-do-campo Lycalopex vetulus (Carnivora, Canidae) em áreas de pastagem e campo sujo, Campinápolis, Mato Grosso, Brasil

Diante da crescente descaracterização do Bioma Cerrado em função da expansão da fronteira agropecuária na região central do Brasil, torna-se importante avaliar a capacidade de adaptação das espécies ao ambiente antropizado. Neste sentido, este trabalho foi desenvolvido com o objetivo de estimar e comparar a densidade populacional da raposa-do-campo Lycalopex vetulus (Lund, 1842) em duas áreas com diferentes graus de alteração, pastagem e campo sujo, em Campinápolis, Mato Grosso. Para tanto, no período entre agosto a novembro de 2005, foram efetuados censos noturnos ao longo de transectos lineares, totalizando percursos de 129,8 km na área de campo sujo e 62,08 km na área de pastagem. Estimativas de densidade populacional foram geradas utilizando o programa Distance 5.0, sendo que o modelo e ajuste mais adequados aos dados foram half-normal + hermite. Foram obtidas 23 e 52 detecções de raposas-do-campo nas áreas de campo sujo e pastagem, respectivamente. A densidade populacional de raposa-do-campo na área de pastagem (D=4,28 indivíduos/km²; IC=2,69 - 6,82) foi maior que na área de campo sujo (D=1,21 indivíduos/km²; IC=0,73 - 2,01), fato que deve estar relacionado, principalmente, com a disponibilidade de alimento e redução de potenciais predadores. Por apresentar uma dieta composta principalmente de cupins, especialmente os dos gêneros Syntermes e Cornitermes, a raposa-do-campo encontra na área de pastagem uma base alimentar abundante e estável. Além disto, a simplificação ambiental, em função da implantação de pastagens acaba por reduzir, ou até mesmo eliminar, animais que são potenciais predadores de raposas-do-campo, como Chrysocyon brachyurus (Illiger, 1815), favorecendo o aumento da densidade populacional da espécie neste tipo de ambiente. Por fim, características adaptativas apresentadas pela raposa-do-campo têm permitido que esta espécie sobreviva, inclusive apresentando elevada densidade populacional, em áreas de pastagem utilizadas para a criação de gado, em Campinápolis, Mato Grosso, onde a vegetação original era Cerrado.

Characterization of Rabies virus isolated from canids and identification of the main wild canid host in Northeastern Brazil

The rabies cases in dogs and wild canids in Northeastern Brazil are a public and animal health problem. This paper describes the identities of the coding region of the N-gene of Rabies virus (RABV) isolated in canids from Northeastern Brazil. The genetic tree generated using the sequence data described here divided the cluster BRAZILAN CANIDS into two subclusters (DOG-RELATED STRAINS and WILD CANID-RELATED STRAINS) with identities greater than those already described. The two subclusters are sub-divided into geographic groups related to the origin of the isolates, suggesting a long-standing ecological coexistence of the sequence types characteristic of the groups.This article also analyzes the 513-nucleotide stretch of the mitochondrial DNA control region of rabies-positive canids from Northeastern Brazil with a view to identifying the main RABV host among them. Among the four species of wild canids found in the region, two (Cerdocyon thous and Pseudalopex vetulus) are frequently associated with rabies. Phylogenetic analysis of sequence data generated from mtDNA suggests that C. thous is the main wild canid host in the region. The results obtained in this study are in concordance with the zoology and ecology of wild canids, and thus, help improve epidemiologic vigilance of rabies and allow a more targeted control of the disease.

Serologic evidence of Leishmania infection in free-ranging wild and domestic canids around a Brazilian National Park

Transmission of disease between wildlife, domestic animals, and humans is of great concern to conservation issues and public health. Here we report on the prevalence of anti-Leishmania sp. antibodies in 21 wild canids (7 Chrysocyon brachyurus, 12 Cerdocyon thous, and 2 Lycalopex vetulus) and 74 free domestic dogs (Canis familiaris) sampled around the Serra do Cipó National Park. In dogs, the apparent prevalence was 8.1% and in wild canids it was 19% (2 crab-eating foxes, C. thous, and 2 maned wolves, C. brachyurus). Management of the domestic dog population with evaluation of incidence changes in humans and wildlife, and enlightenment on the role of wild reservoirs are essential issues for future action and research.

First observations on South America's largely insectivorous canid: the hoary fox (Pseudalopex vetulus)

AbstractThe breeding behaviour and social dynamics of a free‐ranging group of hoary foxesPseudalopex vetuluswere studied in the central cerrado (savannah) region of Brazil. The average adult size was 3.42 kg [95% confidence limits (CL): 2.92–3.91 kg]. The breeding pair occupied overlapping home ranges of 420–456 ha, where they reared a single litter of five offspring (M:F: 3:2) in 12 months. There were no additional adult group members and all offspring dispersed whenc. 10 months old, at least three of which (both sexes) established a home range adjacent to the natal territory. The proportion of time invested in rearing offspring by the breeding female declined significantly (33–2%) with increasing offspring age (pre‐weaning to offspring independence), in contrast to that of the breeding male, which was relatively constant (17–29%); the male acted as a chaperone to foraging offspring, mostly in the absence of the breeding female. Hoary fox diet comprised predominantly of insects, particularly termites and dung beetles, none of which was sufficiently bulky to make carriage to the cubs energetically efficient.