First molecular and serological detection of Epizootic Hemorrhagic Disease virus in white tailed deer ( Odocoileus virginianus ) from Tamaulipas, Mexico / Primeira detecção molecular e sorológica do vírus Doença Hemorrágica Epizoótica em veados de cauda branca (Odocoileus virginianus) de Tamaulipas, Mexico

Epizootic hemorrhagic disease viruses (EHDV) are dsRNA arboviruses transmitted by biting midges of the genus Culicoides that cause disease in domestic and wild ruminants. Epizootic hemorrhagic disease (EHD) is considered the most important infectious disease of white tailed deer (WTD) in North America, some studies in Northeast Mexico reported EHDV-seropositive WTD and EHDV-infected Culicoides vectors. The increasing population of WTD that share habitat with livestock in Northeast México highlights the importance of EHD for the livestock industry in the transboundary region with the U.S. One hundred and twenty two samples from WTD in Tamaulipas state, Mexico were tested by ELISA and RT-PCR for EHDV antibodies and nucleic acid, respectively. Twelve animals were seropositive to ELISA and eleven animals were positive by RT-PCR. This is the first report of EHDV nucleic acid detection in WTD from Mexico. It is hypothesized that applying the transboundary disease approach to interdisciplinary research will help fill knowledge gaps, which could help develop countermeasures to mitigate the threat of EHDV infection in wildlife and livestock along the U.S.-Mexico border.(AU)

Virus da doença hemorrágica epizoótica (EHDV) são arbovírus dsRNA transmitidos por mordidas do genus Culicoides que causam doenças em ruminantes domésticos e selvagens. Doença hemorrágica epizoótica (EHD) é considerada uma das doenças infecciosas mais importantes dos veados de cauda branca (WTD) na América do Norte. Alguns estudos no Nordeste do México relatam soropositividade para EHDV em WTD e vetores Culicoides infectados com EHDV. A crescente população de WTD que compartilham hábitats com pecuária no Nordeste do México realçam a importância de EHD para a indústria pecuária na região de fronteira com os Estados Unidos. Cento e vinte duas amostras de WTD no estado de Tamaulipas, Mexico, foram testados por ELISA e RT-PCR para anticorpos e ácido nucleico de EHDV, respectivamente. Esse é o primeiro relato de detecção de ácido nucleico de EHDV em WTD do México. A hipótese é de que a aplicação de uma resposta transfronteira e pesquisa interdisciplinar ajudará a preencher lacunas de conhecimento levando a medidas reativas para mitigar a ameaça de infecção por EHDV na pecuária e animais selvagens na fronteira entre os Estados Unidos e o Mexico.(AU)

Snakebites as a largely neglected problem in the Brazilian Amazon: highlights of the epidemiological trends in the State of Amazonas

Envenoming snakebites are thought to be a particularly important threat to public health worldwide, especially in rural areas of tropical and subtropical countries. The true magnitude of the public health threat posed by snakebites is unknown, making it difficult for public health officials to optimize prevention and treatment. The objective of this work was to conduct a systematic review of the literature to gather data on snakebite epidemiology in the Amazon region and describe a case series of snakebites from epidemiological surveillance in the State of Amazonas (1974-2012). Only 11 articles regarding snakebites were found. In the State of Amazonas, information regarding incidents involving snakes is scarce. Historical trends show an increasing number of cases after the second half of the 1980s. Snakebites predominated among adults (20-39 years old; 38%), in the male gender (78.9%) and in those living in rural areas (85.6%). The predominant snake envenomation type was bothropic. The incidence reported by the epidemiological surveillance in the State of Amazonas, reaching up to 200 cases/100,000 inhabitants in some areas, is among the highest annual snakebite incidence rates of any region in the world. The majority of the cases were reported in the rainy season with a case-fatality rate of 0.6%. Snakebite envenomation is a great disease burden in the State of Amazonas, representing a challenge for future investigations, including approaches to estimating incidence under-notification and case-fatality rates as well as the factors related to severity and disabilities.

Variabilidad genética en géneros de ciervos neotropicales (Mammalia: Cervidae) según loci microsatelitales

Genetic variability in Neotropical deer genera (Mammalia: Cervidae) according to DNA microsatellite loci. Species conservation programs are highly based on analyses of population genetics. We compared eight Neotropical Cervidae (Mazama americana, M. gouzaoubira, M. rufina, Odocoileus virginianus, Hippocamelus antisensis, Pudu mephistopholes, Ozotoceros bezoarticus and Blastoceros dichotomus) and some European and Asian Cervidae (Cervus elaphus, C. nippon, Capreolus capreolus, C. pygargus and Dama dama). The European species C. elaphus was our standard for a high degree of genetic variability: we used a Scottish population originated in the mix of diverse Western European subspecies. On the contrary, Cervus nippon (a population from Scotland with a founder effect) was our standard for a depauperated population. The M. americana, M. gouzaoubira and O. virginianus samples had high diversity values close to our C. elaphus population (H= 0.64, 0.70 and 0.61, respectively), while M. rufina was very low, close to C. nippon. Several sample sets of Mazama and Odocoileus yielded a homozygote excess, probably due to the Wahlund (subdivison) effect. There was no evidence of recent bottleneck events. Rev. Biol. Trop. 57 (3): 879-904. Epub 2009 September 30.

Los programas de conservación de especies se apoyan fuertemente en estudios de genética poblacional. En el presente estudio, reportamos diversos análisis genéticopoblacionales en ocho especies de cérvidos neotropicales (Mazama americana, M. gouzaoubira, M. rufina, Odocoileus virginianus, Hippocamelus antisensis, Pudu mephistopholes, Ozotoceros bezoarticus y Blastoceros dichotomus) y, adicionalmente, en varias especies de cérvidos europeos y asiáticos (Cervus elaphus, C. nippon, Capreolus capreolus, C. pygargus and Dama dama). Una de esas especies europeas, la población de Cervus elaphus en Escocia, fue tomada como una población con un grado muy elevado de diversidad genética ya que proviene del cruce de diferentes grupos de ciervos rojos procedentes de diversas subespecies de la Europa continental. Desde una perspectiva de una diversidad genética depauperada, se tomó el nivel encontrado en una población de ciervos sika (Cervus nippon) en Escocia, que prácticamente no mostró variabilidad a nivel molecular. Respecto a esos dos casos que consideramos como de elevada y escasa variabilidad genética, encontramos que las poblaciones analizadas de Mazama americana, M. gouzaoubira y Odocoileus virginianus estuvieron cerca del límite máximo encontrado para el ciervo rojo escocés (H=0.64, 0.70 y 0.61, respectivamente), mientras que M. rufina mostró el más bajo grado de variabilidad genética de las especies neotropicales, cercano al extremo mínimo presentado por C. nippon. Algunas de las muestras de Mazama y de Odocoileus, tomadas a nivel macrogeográfico, mostraron un exceso de homocigotos debido, probablemente, a la existencia de efecto Wahlund (efecto de subdivisión). Ninguna de las especies analizadas parece haber atravesado un cuello de botella reciente.

Genetic variability of the prion protein gene (PRNP) in wild ruminants from Italy and Scotland

The genetics of the prion protein gene (PRNP) play a crucial role in determining the relative susceptibility to transmissible spongiform encephalopathies (TSEs) in several mammalian species. To determine the PRNP gene variability in European red deer (Cervus elaphus), roe deer (Capreolus capreolus) and chamois (Rupicapra rupicapra), the PRNP open reading frame from 715 samples was analysed to reveal a total of ten single nucleotide polymorphisms (SNPs). In red deer, SNPs were found in codons 15, 21, 59, 78, 79, 98, 136, 168 and 226. These polymorphisms give rise to 12 haplotypes, and one of which is identical to the PRNP of American wapiti (Rocky Mountain elk, Cervus elaphus nelsoni). One silent mutation at codon 119 was detected in chamois and no SNPs were found in roe deer. This analysis confirmed that European wild ruminants have a PRNP genetic background that is compatible with TSE susceptibility, including chronic wasting disease.

Chromosome polymorphism in the Brazilian dwarf brocket deer, Mazama nana (Mammalia, Cervidae)

The Brazilian dwarf brocket deer (Mazama nana) is the smallest deer species in Brazil and is considered threatened due to the reduction and alteration of its habitat, the Atlantic Rainforest. Moreover, previous work suggested the presence of intraspecific chromosome polymorphisms which may contribute to further population instability because of the reduced fertility arising from the deleterious effects of chromosome rearrangements during meiosis. We used G- and C-banding, and nucleolus organizer regions localization by silver-nitrate staining (Ag-NOR) to investigate the causes of this variation. Mazama nana exhibited eight different karyotypes (2n = 36 through 39 and FN = 58) resulting from centric fusions and from inter and intraindividual variation in the number of B chromosomes (one to six). Most of the animals were heterozygous for a single fusion, suggesting one or several of the following: a) genetic instability in a species that has not reached its optimal karyotypic evolutionary state yet; b) negative selective pressure acting on accumulated rearrangements; and c) probable positive selection pressure for heterozygous individuals which maintains the polymorphism in the population (in contrast with the negative selection for many rearrangements within a single individual).

Genetic diversity of two Brazilian populations of the Pampas deer (Ozotoceros bezoarticus, Linnaeus 1758)

The Pampas deer (Ozotoceros bezoarticus) is one of the most endangered Neotropical cervid with populations that have been drastically reduced to small and isolated ones, mainly because of its habitat destruction. Random amplified polymorphic DNA (RAPD) markers were used to analyze population divergence and genetic variation within and between two populations corresponding to distinct subspecies. The RAPD markers displayed substantial genetic variation with all animals possessing unique RAPD phenotypes over 105 polymorphic bands produced by 15 primers. An analysis of molecular variance (AMOVA) and a neighbor-joining cluster analysis were performed to assess levels of differentiation between populations. No differentiation was recorded and about 96.0 percent (P < 0.00001) of the total variance was attributable to variation within populations. This result is quite distinct from data obtained by the analysis of the mtDNA control region, and is discussed on the basis of genetic differences between the different markers and the male-biased dispersal patterns generally observed in the mammal species. The data presented herein are potentially useful for future taxonomic and genetic studies in this species, for the monitoring of the genetic variation observed within these populations, and for the development of management guidelines for its conservation.

O Veado-campeiro (Ozotoceros bezoarticus) é uma das espécies de cervídeos neotropical mais ameaçadas devido à destruição de seu hábitat e conseqüente redução e isolamento de suas populações. Marcadores do tipo RAPD (Random amplified polymorphic DNA) foram utilizados na análise da divergência populacional e estimativa da variação genética dentro e entre duas populações correspondentes a diferentes subespécies. Os marcadores RAPD mostraram uma variação genética substancial, sendo que as 105 bandas polimórficas obtidas pelo uso de 15 primers produziram fenótipos únicos para todos os indivíduos analisados. Para avaliar o nível de diferenciação entre as populações, foi realizada uma análise da variância molecular (AMOVA) e uma análise de agrupamento utilizando o método de neighbor-joining. Nenhuma diferenciação foi observada, sendo aproximadamente 96,0 por cento da variação encontrada atribuída à variação dentro das populações estudadas. Este resultado difere do obtido através da análise da região controle do mtDNA, e é discutido levando-se em consideração as diferenças genéticas entre os diferentes marcadores utilizados e o padrão de dispersão geralmente observado nas espécies de mamíferos (realizada principalmente pelos machos). Os dados aqui apresentados poderão ser úteis para futuros estudos taxonômicos e genéticos desta espécie, para o monitoramento da variação genética observada em suas populações e para o desenvolvimento de estratégias de manejo para sua conservação.

Relaciones filogenéticas entre géneros de ciervos neotropicales (Artiodactyla: Cervidae) mediante secuenciación de ADN mitocondrial y marcadores microsatelitales

Phylogenetic relationships among Neotropical deer genera (Artiodactyla: Cervidae) by means of DNAmt sequences and microsatellite markers. The current work shows two molecular phylogenetic analyses on Neotropical deers. In the first analysis, the mitochondrial control region (D-loop) was sequenced in six Odocoileinae species from Latin America, using the sequences of two Muntiacinae as outgroups. The results obtained were as follows: A sequence of Mazama americana showed a striking relationship with several sequences of Odocoileus in contrast to that expected, since this M. americana haplotype, from a Mexican origin, was not associated with several Bolivian Mazama sequences analyzed. This could put forward that this genera is not monophyletic. On the other hand, these Bolivian Mazama formed a clade with Pudu puda and Ozotoceros bezoarticus. Likely, an Odocoileus virginianus sequence from the Central area of Colombia showed a more strong relationship with a Northamerican O. heminonus sequence than with the other O. virginianus sequences of Colombian origin as well. This could be explained by means of various different hypotheses. The first is the existence of common ancestral haplotypes between both species. Another one is the reiterative hybridization among both Odocoileus species before the migration of O. virginianus from North America to South America. Moreover, the maximum parsimony analysis showed an intense relationship between the Muntiacinae and this Neotropical Cervidae clade. In addition, and adding credence to the relevant polyphyletism found in Mazama by means of the mitochondrial control region DNA sequences, a second analysis with 16 DNA microsatellite loci also showed a higher genetic relationship between M. americana and O. virginianus, than between the first species regard to Mazama gouazoubira. Rev. Biol. Trop. 55 (2): 723-741. Epub 2007 June, 29.

El presente trabajo muestra dos análisis moleculares sobre la filogenia de los cérvidos neotropicales. En uno se secuenció la región control del mtDNA (D-loop) de seis especies de Odocoileinae, utilizándose las secuencias de dos Muntiacinae como elementos externos. Se evidenciaron los siguientes resultados: La secuencia de una Mazama americana, de origen mexicano, presentó una fuerte relación filogenética con las diversas secuencias estudiadas de Odocoileus, contrario a lo esperado ya que, a priori, debería haberse asociado con las secuencias analizadas de otros ejemplares de Mazama de origen boliviano. Esto pone en evidencia que este género no es monofilético. A su vez, las secuencias de los ejemplares bolivianos de Mazama formaron una agrupación con secuencias de Pudu puda y O. bezoarticus. Una secuencia de O. virginianus, del área central de Colombia, presentó más relación con la secuencia de un O. hemionus norteamericano que con las restantes secuencias analizadas de O. virginianus, también de origen colombiano. Esto puede reflejar varias explicaciones hipotéticas, tales como la existencia de haplotipos ancestrales comunes entre ambas especies de Odocoileus, hasta la hibridación en Norteamérica entre ambos taxones antes de su penetración en Sudamérica. Los análisis de máxima parsimonia presentan una especial relación entre los Muntiacinae y el clado de los ciervos sudamericanos. El segundo análisis filogenético hizo uso de 16 marcadores nucleares microsatélites. Aunque, en principio, estos marcadores no son los más recomendables para estudios filogenéticos intergenéricos, los resultados muestran, al igual que el ADN mitocondrial, una mayor relación entre M. americana y O. virginianus que entre la primera especie y M. gouzaoubira.

Cross-amplification tests of ungulate primers in the endangered Neotropical pampas deer (Ozotoceros bezoarticus)

In cross-species amplification tests of 15 ungulate primers in pampas deer, five were retained to form a small panel of highly polymorphic loci that could be used to efficiently screen populations of this endangered species. The polymerase chain reactions were performed incorporating the universal fluorescent labeled M13 (-21) primer. In 69 pampas deer, average allelic diversity was 15, expected heterozygosity was 0.869 and the mean polymorphic information content value was 0.847. Paternity exclusion probabilities over loci were NE-1P = 0.01336 and NE-2P = 0.00135, and combined non-exclusion probability of identity was P(ID) = 3 × 10-8.

Transferability of microsatellite loci from Cervidae species to the endangered Brazilian marsh deer, Blastocerus dichotomus

Blastocerus dichotomus, the marsh deer, is the largest Brazilian Cervidae species. The species is endangered because of hunting and loss of its natural habitat, i.e., flood plain areas, because of hydroelectric power station construction and agricultural land expansion. In the present study, we tested 38 microsatellite loci from four Cervidae species: Odocoileus virginianus (7), Rangifer tarandus (17), Capreolus capreolus (7), and Mazama bororo (7). Eleven loci showed clear amplification, opening a new perspective for the generation of fundamental population genetic data for devising conservation strategies for B. dichotomus.

Identification of single-nucleotide polymorphisms of the prion protein gene in sika deer (Cervus nippon laiouanus)

Polymorphisms of the prion protein gene (PRNP) havebeen detected in several cervid species. In order toconfirm the genetic variations, this study examined theDNA sequences of the PRNP obtained from 33 captivesika deer (Cervus nippon laiouanus) in Korea. A total ofthree single-nucleotide polymorphisms (SNPs) at codons100, 136 and 226 in the PRNP of the sika deer wereidentified. The polymorphic site located at codon 100 hasnot been reported. The SNPs detected at codons 100 and226 induced amino acid substitutions. The SNP at codon136 was a silent mutation that does not induce any aminoacid change. The genotype and allele frequencies weredetermined for each of the SNPs.

Obtaiment of Pudu (Pudu pudu) deer embryos by the somatic nuclear transfer technique

RESUMEN: Los objetivos de este trabajo fueron los de producir embriones de pudú, obtenidos por la transferencia de núcleos de fibroblastos de la oreja de pudú en ovocitos de un rumiante domésticos que es el bovino. Para posteriormente en un trabajo futuro proceder a la transferencia de embriones de pudú, al útero de hembras receptoras sincronizadas de otra especie. Se obtuvieron biopsias de 1 mm aproximadamente del borde externo de la orejas de dos ciervos pudu machos del jardín zoológico Buin-Zoo, Santiago de Chile. Las líneas celulares han sido establecidas y conservadas según los protocolos utilizados para las bovinos. Los ovocitos son obtenidos por punción del complejo cúmulos-ovocito (COC).desde ovarios de vacas recuperados del matadero. Cada ovocito es enucleado y fusionado con un fibroblasto aislado insertado bajo la zona pelúcida. La fusión de membranas celulares es obtenida por choques eléctricos. En cuanto a la cronología, observamos que al segundo día se forma una etapa de dos blastómeras, al tercer día mórulas de 8 a 16 células, y desde el cuarto día se ha diferenciado como blastocisto, el cuál al séptimo día termina por eclosionar de la zona pelúcida.La obtención de blastocistos embrionarios indica que es posible obtener embriones de pudú mediante clonaje heteroespecífico, aunque, el porcentaje de éxito obtenido es relativamente bajo. Queda aun por verificar la viabilidad de los embriones así obtenidos después de la transferencia in útero.

Caracterización morfométrica de cuatro subespecies de venado cola blanca (Odocoileus virginianus) en la zona noreste de México / Morphometric charaterization of four white-tailed deer subspecies (Odocoileus virginianus) in the northeas region of Mexico

Con el objetivo de contribuir en la definición de las subespecies de venado cola blanca de México, se determinó la variabilidad morfométrica de las subespecies (Odocoileus virginianus carminis, O.v. miquihuanensis, O.v. texanus y O.v. veraecrucis) que se distribuyen en el Noreste de México. Se analizó la información de 592 individuos (579 O. v. texanus, seis O.v. veraecrucis, cuatro O.v. miquihuanesis y tres O.v. carminis), y 49 cráneos de O.v. texanus. La comparación entre la morfometría histórica y la del presente estudio mostró un 98% de similitud para O.v carminis, O.v. miquihuanensis y O.v veraecrucis y un 96% para O.v. texanus. Los contrastes otorgonales mostraron diferencias morfométricas (P<0,05) entre las subespecies. En O.v. texanus, la longitud total tendió a ser de mayor tamaño que en las otras subespecies, mientras que O.v. veraecrucis mostró diferencias significativas respecto a O.v. carminis y O.v. miquihuanesis en todas las variables. La mayor similitud por Distancia Euclidiana se presentó entre los machos de O.v. carminis y O.v. miquihuanensis (5,1), mientras que la de menor similitud fue entre los machos de O.v. texanus y los de las otras tres subespecies (19,9). Los machos adultos de Tamaulipas se diferenciaron significativamente (P<0,05) de los de Coahuila y Nuevo León.