Ticks associated with armadillo (Euphractus sexcinctus) and anteater (Myrmecophaga tridactyla) of Emas National Park, State of Goias, Brazil

This study was conducted in October 1998 and November 1999 in the Emas National Park (131,868 ha), a savanna-type cerrado region situated in the far south of Goias State, Brazil, near the geographic center of South America (15degrees-23degrees S; 45degrees-55degrees W). Animals were captured with the aid of nets and anesthetized (15 mg/kg ketamine + 1 mg/kg xylasine) in order to collect ticks for identification and to establish laboratory colonies. They included giant anteaters (Myrmecophaga tridactyla) (n = 4) and yellow armadillos (Euphractus sexcinctus) (n = 6). Free-living ticks (larvae, nymphs, and adults) were collected from the field by using a 1 X 2-m flannel cloth. Free-living ticks were identified as Amblyomma sp., A. cajennense, and A. triste. Adult ticks collected from anteaters were identified as Amblyomma cajennense and A. nodosum and from armadillos as A. pseudoconcolor and A. nodosum. The relevance of these host-tick relationships to possible mechanisms underlying emergence of tick-borne pathogens of importance to public health is discussed.

Filogenia molecular dos xenarthra (Mammalia): análise do grupo cingulata a partir de sequências nucleotídicas do gene mitocondrial rRNA 16S

Os Xenarthra são o grupo de mamíferos que inclui os tatus, os tamanduás e as preguiças. A América do Sul serviu de cenário para a história natural do grupo que, somente no fim do Cenozóico, dispersou-se para a América Central e, com uma perda de variedade, chegou à América do Norte e à algu-mas ilhas do Caribe. Trinta e uma espécies estão descritas dentro da linha-gem dos Xenarthra. Elas estão classificadas em 13 gêneros, quatro famílias (Bradypodidae, Megalonychidae, Myrmecophagidae e Dasypodidae) e duas ordens (Cingulata e Pilosa). A filogenia deste grupo tem sido alvo de diver-sas pesquisas que analisaram tanto dados morfológicos, quanto moleculares. Delsuc et al. (2003) analisaram seqüências de genes mitocondriais e nucleares e confirmaram a monofilia das três subfamílias (Dasypodinae, Euphacti-nae e Tolypeutinae) inclusas na família Dasypodidae. Delsuc et al. (2003) geraram a seguinte árvore: (((Bradypus, Choloepus)100, ((Myrmecophaga, Tamandua)100, Cyclopes)100), ((D. kappleri, D. novemcinctus)100, (Toly-pentes, (Priodontes, Cabassous)54)100, (Zaedyus, (Euphractus, Chaetophrac-tus)60)100)). Gaudin (2005) apresentou um trabalho que reviu e ampliou as análises morfológicas apresentadas até então, concluindo que os tatus atu-ais estão divididos em dois grupos, um mais basal (Dasypodinae) e outro mais derivado (Euphractinae), de acordo com o seguinte arranjo: (Bradypus, Tamandua), (Dasypus, (Priodontes, (Cabassous, (Tolypeutes, (Euphractus, Chaetophractus, (Zaedyus, Chlamyphorus)42)36)72)72)40)85). Neste traba-lho utilizou-se parte do gene mitocondrial rRNA 16S de 12 táxons atu-ais de Xenarthra para analisar a filogenia do grupo através do critério de máxima verossimilhança. Nossos resultados são apresentados analisando-se o gene 16S e analisando o banco de dados do 16S mais o de Delsuc et al. (2003). Nas duas situações, as filogenias apresentadas apóiam os resulta-dos de Delsuc et al. (2003): (Bradypus, (Choloepus, ((Cyclopes, (Myrme-cophaga, Tamandua)100)100, (Dasypus, (((Cabassous, Priodontes)68, Toly-peutes)100,((Chaetophractus, Euphractus)65, Zaedyus)100)100)100)100)100). Uma melhora nos valores de bootstrap nos ramos dentro das sub-famílias da família Dasypodidae é percebida em relação ao trabalho de Delsuc et al. (2003). Acreditamos que Elementos de Transposição do tipo (LINES) são os marcadores moleculares mais adequados para confirmar o arranjo obtido com as seqüências de genes mitocondriais e nucleares.

MANAGEMENT OF A CARAPACE FRACTURE IN A SIX-BANDED ARMADILLO (EUPHRACTUS SEXCINCTUS)

An adult female free-ranging six-banded armadillo (Euphractus sexcinctus) was presented with an inverted L-shaped fracture of the left pectoral carapace. The fracture was stabilized with the use of three simple interrupted interfragmentary sutures of 2-0 nylon. After 7 days, wound dehiscence occurred, so sutures were replaced and the wound treated topically with castor bean oil cream. Healing of the fracture was observed after 14 days of this treatment.

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)

Armadillo

Oil 24 x 24"

Identification of Darlin, a Dictyostelium Protein with Armadillo-like Repeats that Binds to Small GTPases and Is Important for the Proper Aggregation of Developing Cells

We purified from Dictyostelium lysates an 88-kDa protein that bound to a subset of small GTPases, including racE, racC, cdc42Hs, and TC4ran, but did not bind to R-ras or rabB. Cloning of the gene encoding this 88-kDa protein revealed that it contained multiple armadillo-like repeats most closely related to the mammalian GTP exchange factor smgGDS. We named this protein darlin (Dictyostelium armadillo-like protein). Disruption of the gene encoding darlin demonstrated that this protein is not essential for cytokinesis, pinocytosis, phagocytosis, or development. However, the ability of darlin null cells to aggregate in response to starvation is severely affected. When starved under liquid medium, the mutant cells were unable to form aggregation centers and streams, possibly because of a defect in cAMP relay signaling. This defect was not due to an inability of the darlin mutants to activate adenylate cyclase in response to G protein stimulation. These results suggest that the darlin protein is involved in a signaling pathway that may modulate the chemotactic response during early development.

RcppArmadillo : accelerating R with high-performance C++ linear algebra

The R statistical environment and language has demonstrated particular strengths for interactive development of statistical algorithms, as well as data modelling and visualisation. Its current implementation has an interpreter at its core which may result in a performance penalty in comparison to directly executing user algorithms in the native machine code of the host CPU. In contrast, the C++ language has no built-in visualisation capabilities, handling of linear algebra or even basic statistical algorithms; however, user programs are converted to high-performance machine code, ahead of execution. A new method avoids possible speed penalties in R by using the Rcpp extension package in conjunction with the Armadillo C++ matrix library. In addition to the inherent performance advantages of compiled code, Armadillo provides an easy-to-use template-based meta-programming framework, allowing the automatic pooling of several linear algebra operations into one, which in turn can lead to further speedups. With the aid of Rcpp and Armadillo, conversion of linear algebra centered algorithms from R to C++ becomes straightforward. The algorithms retains the overall structure as well as readability, all while maintaining a bidirectional link with the host R environment. Empirical timing comparisons of R and C++ implementations of a Kalman filtering algorithm indicate a speedup of several orders of magnitude.

Monitoreo de la fauna silvestre en bosque tropical seco secundario, Nandarola, Nandaime, Nicaragua

Los animales silvestres obtienen refugio, hábitat, alimento y agua de la cobertura que les ofrece la vegetación dentro del bosque. En los años 60s, la mayoría de las áreas boscosas de Nandarola fueron deforestadas y usadas en la producción agrícola de granos (frijoles, maíz, arroz) y áreas ganaderas. Los campos fueron recientemente abandonados y están siendo colonizados por un joven bosque secundario (González, 2005). La fauna silvestre también sufre procesos de colonización de las áreas abandonadas, de esta forma, es importante conocer que especies son las primeras en ser parte de esa colonización y evaluar los cambios en composición de las comunidades animales en el tiempo, principalmente porque muchos vegetales dependen de éstos para realizar procesos biológicos y ecológicos tan importantes como lo son la dispersión y la polinización. Hasta el momento se han realizados dos muestreos para el monitoreo de la fauna silvestre en un parche de bosque seco secundario de 3.5 ha en Nandarola, realizándose 17 visitas en el primero (año 2007) y 15 en el segundo (año 2009), distribuidas en ocho meses (octubre-mayo en cada muestreo). El área era agrícola y fue abandonado hace 17 años con fines de convertirlo en área forestal. Para la toma de datos se establecieron seis transectos de 20 x 100 m y seis puntos de conteo de 25 m de radio. Fueron contados e identificados todos los animales silvestres de los grupos mamíferos, aves, reptiles y anfibios mediante la técnica de avistamiento, para los cuales se calculó, se comparó la riqueza, abundancia, diversidad biológica y se describió la composición de especie, la preferencia de hábitat, los hábitos alimenticios y también se determinaron las especies de probable ocurrencia en el área de estudio. En promedio entre los dos muestreos fueron determinaron 113 individuos, agrupados en 35 especies, 34 géneros y 26 familias de fauna silvestre. De estas especies cuatro fueron mamíferos, 21 aves, siete reptiles y tres anfibios. Las familias más representadas según el número de especies fueron: Columbidae (3 especies) e Iguanidae (3). Las especies con mayor promedio de abundancia fueron: Campylorynchus rufinucha (22.5 individuos), Thryothorus pleurostictus (11.5), Calocitta formosa (10.5), Turdus grayi (8), Pachyrampuhus aglaiae (7), Eumomota supersiliosa (6.5) y Trogon melanocephalus (6). La abundancia y la riqueza no varió significativamente entre los muestreos, la diversidad fue mayor en el primero, debido al aumento de la dominancia específica (principalmente por Campylorynchus rufinucha ) en el segundo muestreo. El grupo de los mamíferos, reptiles y anfibios fue poco representado en el área de estudio, pudiéndose deber a factores antropogénicos. El paisaje esta mayormente representado por especies de aves de hábitats abiertos y que se alimentan mayoritariamente de invertebrados y semillas. Por otro lado con una relación menos específica los mamíferos y reptiles fueron principalmente de hábitats generalistas y mayormente consumidores de frutas, hierbas y vertebrados. Entre las especies de probable ocurrencia en el área de estudio se determinaron: Bufo coccifer, Boa constrictor, Ctenosaura similis, Mabuya unimarginata, Amazilia rutila, Aratinga canicularis, Ciccaba virgata, Crax rubra, Polyborus plancus, Dasypus novencintus, Dasyprocta punctata, Felis pardalis, Felis wiedii, Philander opossum, Nasua narica, Odocoileus virginianus, Procyon lotor, Puma concolor, Sylvilagus sp, Tayassu tajacu, entre otros.

A influência de uma espécie exótica invasora, Artocarpus heterophyllus Lam. (jaqueira), sobre uma comunidade de pequenos mamíferos e sua interferência na dinâmica de dispersão de sementes nativas

Atualmente, sabe-se que danos causados por espécies exóticas invasoras são umas das principais causas de extinção de espécies, afetando mais seriamente espécies que evoluíram em ilhas. A jaqueira, Artocarpus heterophyllus Lamarck (Moraceae) é originária das florestas tropicais da Índia. Foi introduzida no Brasil ainda no período Colonial e atualmente é invasora em áreas de Mata Atlântica, incluindo a Ilha Grande, RJ. Durante três anos foram amostrados bimestralmente 18 grades, 10 com diferentes densidades de jaqueiras e oito sem jaqueiras. Em cada grade foram colocadas 11 armadilhas de captura de mamíferos, que ficavam abertas durante três dias consecutivos por mês. No laboratório as fezes de todos os animais capturados foram analisadas para verificar a dieta e a quantidade de sementes nativas defecadas. Para verificar as espécies capazes de predar e dispersar sementes de jaca, fizemos testes com sementes de jaca atados a carretéis e armadilhas fotográficas. Neste contexto, o estudo teve como objetivo verificar a influência da jaqueira na comunidade de pequenos mamíferos e na dispersão de sementes de espécies nativas. Os resultados mostraram que em áreas com maior densidade de jaqueiras adultas, houve uma maior abundância de espécies frugívoras e a diminuição da abundância de espécies mais insetívoras. Embora a jaqueira não tenha influenciado no consumo de itens de origem animal e vegetal entre áreas com e sem jaqueiras e durante os períodos de maior e menor frutificação, essa espécie desfavoreceu a dispersão de sementes nativas. Em áreas com maior densidade de jaqueiras verificamos uma quantidade menor de sementes nativas sendo defecadas pelos pequenos mamíferos. O número de sementes defecadas durante o período de menor frutificação das jaqueiras não foi significativo em relação ao período de maior frutificação e em todos os períodos somados. Já em relação à frequência de fezes contendo sementes nativas, os resultados das regressões simples foram significativos para todos os períodos. O fruto da jaqueira A. heterophyllus foi mais consumido por D. aurita, T. dimidiatus eCuniculus paca, sendo que D. auritanão teve influência sobre a predação e a dispersão de sementes de jaca. Os roedores T. dimidiatuse C. paca foram registrados pelas armadilhas fotográficas predando 20% e 16% das sementes de jaca e carregaram 65% e 44% das sementes, respectivamente. Os testes com carretel mostraram que 86% das sementes foram predadas, 10% foram deixadas intactas no local e apenas 4% foram dispersas a pequenas distâncias, entre 2 e 15 metros, sendo possível que esses roedores propiciem a dispersão dessa espécie exótica e invasora para novas áreas

RNAi suppression of zebrafish peptidoglycan recognition protein 6 (zfPGRP6) mediated differentially expressed genes involved in Toll-like receptor signaling pathway and caused increased susceptibility to Flavobacterium columnare

In Drosophila, Toll signaling cascade, which resembles the mammalian Toll-like receptor (TLR)/IL-1R signaling pathways and regulates the expression of anti-microbial peptide genes, mainly relies on peptidoglycan recognition proteins (PGRPs) for the detection of bacterial pathogens. To explore the effect of zebrafish peptidoglycan recognition protein 6 (zfPGRP6) on Toll-like receptor signaling pathway, RNA interference (siRNA) and real time quantitative PCR (RQ-PCR) methods were used to identify differentially expressed genes regulated by zfPGRP6. The target genes included TLR2, TLR3, TLR5, TLR7, TLR8, IL1R, Sterile-alpha and Armadillo motif containing protein (SARM), myeloid differentiation factor 88 (MyD88) and nuclear factor (NF)-kappa B2 (p100/p52). The results of RQ-PCR showed that RNAi-mediated Suppression of zfPGRP6 significantly down-regulated the expression of TLR2, TLR5, IL1R, SARM, MyD88 and p100/p52. The expression of beta-defensin-1 was also down-regulated in those embryos silenced by zfPGRP6. In challenge experiments to determine the anti-bacterial response to Gram-negative bacteria, RNAi knock-down of zfPGRP6 markedly increased susceptibility to Flavobacterium columnare. (C) 2008 Elsevier B.V. All rights reserved.

麂属动物以及六带犰狳DNA重复序列家族的克隆、序列分析和染色体定位

1．黑麂和费氏麂卫星DNA的克隆、序列分析和染色体定位 麂属动物在很短的时间内经历了快速的物种辐射，并且种间染色体数目存在巨大差异，是研究动物核型进化和物种起源的理想模型。近二十年来的分子细胞遗传学研究已基本上证实染色体串联融合(端粒－着丝粒融合)是麂属动物核型演化的主要染色体重排方式。尽管染色体串联融合的分子机制仍不清楚，但研究提示着丝粒区域的卫星DNA可能介导染色体的非同源重组。因此，着丝粒卫星DNA的克隆、分析序列以及染色体定位研究不仅有助于阐明麂属染色体核型演化规律，还可能揭示染色体串联融合的分子机制。迄今为止，上述研究工作已经在赤麂、小麂和小麂台湾亚种开展过。但是，尚无有关黑麂、费氏麂和贡山麂卫星 DNA 克隆、序列分析以及染色体定位研究的报道。 在本研究中，我成功地克隆了黑麂的卫星DNA I、II和IV，分别命名为BMC5、BM700和BM1.1k，并且从费氏麂中克隆了卫星DNA II，命名为FM700。对这些卫星DNA克隆进行序列分析，并将这些克隆探针分别与黑麂、费氏麂、贡山麂和小麂的染色体杂交。研究结果表明： 1)黑麂的卫星DNA I(BMC5)与小麂卫星DNA I(C5)序列高度相似，并且在小麂、黑麂、费氏麂和贡山麂染色体上的大部分串联融合位点处均有分布，因此卫星DNA I可能代表着染色体发生串联融合后保存下来，来源于麂属动物祖先染色体着丝粒的一种卫星DNA。卫星DNA I在这四种麂属动物染色体上的分布也表明黑麂、费氏麂和贡山麂与赤麂的核型演化过程相似，很可能从一个2n ＝ 70的共同祖先通过一系列的串联易位进化而来。 2) 将卫星DNA II（BM700和FM700）克隆探针分别杂交到黑麂和费氏麂的染色体上，只检测到几对间隔分布的信号。这提示在核型进化过程中不同卫星DNA间可能发生了广泛的重组，从而导致卫星DNA II大量丢失。大部分重组断裂位点可能位于卫星DNA I 与卫星DNA II之间，或者在卫星DNA II 区域内。 2．六带犰狳重复序列家族的克隆、序列分析和染色体定位 六带犰狳属于犰狳科、贫齿目，是六带犰狳属中唯一的一个代表物种。系统发育研究认为贫齿目与非洲兽总目是有胎盘哺乳动物中最原始的两个类群。C显带结果揭示六带犰狳30%的基因组是由组成性异染色质构成的，并且C带分布的位置也较复杂，提示在六带犰狳基因组中存在多种重复序列元件。 为了研究六带犰狳异染色质的组成，我从六带犰狳的基因组中克隆了七种位点特异性的重复序列。根据测序结果以及它们在染色体上的分布，将这些重复序列分为五个重复序列家族。其中AMD-EcoRI 837与AMD-BglII 811的序列相似，都是由大小约116 bp的单位组成，分布在大多数染色体的着丝粒区域，同时在一些染色体臂也有分布。AMD-EcoRI 832，AMD-EcoRI 836和AMD-EcoRI 934是特定染色体的重复序列，并且都分布于着丝粒区域。另外，AMD-BglII 634，AMD-EcoRI 731两个克隆都属于长散在分布重复序列(L1)，倾向于分布在G带阳性、富含AT碱基的区域，并且这两种重复序列在染色体上的定位与C带阳性的非着丝粒的异染色质区域很相似。本研究提供了六带犰狳异染色质区域的部分基因组信息，并且这些重复序列家族也可以用于研究六带犰狳及其近缘物种的系统发育关系。

Cytokinesis proteins Tum and Pav have a nuclear role in Wnt regulation.

Wg/Wnt signals specify cell fates in both invertebrate and vertebrate embryos and maintain stem-cell populations in many adult tissues. Deregulation of the Wnt pathway can transform cells to a proliferative fate, leading to cancer. We have discovered that two Drosophila proteins that are crucial for cytokinesis have a second, largely independent, role in restricting activity of the Wnt pathway. The fly homolog of RacGAP1, Tumbleweed (Tum)/RacGAP50C, and its binding partner, the kinesin-like protein Pavarotti (Pav), negatively regulate Wnt activity in fly embryos and in cultured mammalian cells. Unlike many known regulators of the Wnt pathway, these molecules do not affect stabilization of Arm/beta-catenin (betacat), the principal effector molecule in Wnt signal transduction. Rather, they appear to act downstream of betacat stabilization to control target-gene transcription. Both Tum and Pav accumulate in the nuclei of interphase cells, a location that is spatially distinct from their cleavage-furrow localization during cytokinesis. We show that this nuclear localization is essential for their role in Wnt regulation. Thus, we have identified two modulators of the Wnt pathway that have shared functions in cell division, which hints at a possible link between cytokinesis and Wnt activity during tumorigenesis.

Role of the different actin genes during extrusion of capping protein mutant cells in the Drosophila wing blade epithelium

Dissertação mest., Biotecnologia, Universidade do Algarve, 2008

Dominant β-catenin mutations cause intellectual disability with recognizable syndromic features

The recent identification of multiple dominant mutations in the gene encoding β-catenin in both humans and mice has enabled exploration of the molecular and cellular basis of β-catenin function in cognitive impairment. In humans, β-catenin mutations that cause a spectrum of neurodevelopmental disorders have been identified. We identified de novo β-catenin mutations in patients with intellectual disability, carefully characterized their phenotypes, and were able to define a recognizable intellectual disability syndrome. In parallel, characterization of a chemically mutagenized mouse line that displays features similar to those of human patients with β-catenin mutations enabled us to investigate the consequences of β-catenin dysfunction through development and into adulthood. The mouse mutant, designated batface (Bfc), carries a Thr653Lys substitution in the C-terminal armadillo repeat of β-catenin and displayed a reduced affinity for membrane-associated cadherins. In association with this decreased cadherin interaction, we found that the mutation results in decreased intrahemispheric connections, with deficits in dendritic branching, long-term potentiation, and cognitive function. Our study provides in vivo evidence that dominant mutations in β-catenin underlie losses in its adhesion-related functions, which leads to severe consequences, including intellectual disability, childhood hypotonia, progressive spasticity of lower limbs, and abnormal craniofacial features in adults

Comparative phylogeography of Trypanosoma cruzi TCIIc: New hosts, association with terrestrial ecotopes, and spatial clustering

We characterized 28 new isolates of Trypanosoma cruzi IIc (TCIIc) of mammals and triatomines from Northern to Southern Brazil, confirming the widespread distribution of this lineage. Phylogenetic analyses using cytochrome b and SSU rDNA sequences clearly separated TCIIc from TCIIa according to terrestrial and arboreal ecotopes of their preferential mammalian hosts and vectors. TCIIc was more closely related to TCIId/e, followed by TCIIa, and separated by large distances from TCIIb and TCI. Despite being indistinguishable by traditional genotyping and generally being assigned to Z3, we provide evidence that TCIIa from South America and TCIIa from North America correspond to independent lineages that circulate in distinct hosts and ecological niches. Armadillos, terrestrial didelphids and rodents, and domestic dogs were found infected by TCIIc in Brazil. We believe that, in Brazil, this is the first description of TCIIc from rodents and domestic dogs. Terrestrial triatomines of genera Panstrongylus and Triatoma were confirmed as vectors of TCIIc. Together, habitat, mammalian host and vector association corroborated the link between TCIIc and terrestrial transmission cycles/ecological niches. Analysis of ITS1 rDNA sequences disclosed clusters of TCIIc isolates in accordance with their geographic origin, independent of their host species. (C) 2009 Elsevier B.V. All rights reserved.