Susceptibility of Rhipicephalus (Boophilus) microplus to ivermectin (200, 500 and 630 mu g/kg) in field studies in Brazil

The present study aimed to determine the susceptibility of 17 Rhipicephalus (Boophilus) microplus populations, originating in the Southeast and Southern regions of Brazil, to different ivermectin concentrations (200, 500 and 630 mu g/kg), administered through subcutaneous or topical (pour-on) routes. R. (B.) microplus populations from the states of Minas Gerais (seven populations), Sao Paulo (seven populations) and Parana (three populations) were chosen for the tests. The selected cattle were allocated to treatment groups on day 0, and block formation was based on the arithmetic mean of female ticks (4.5-8.0 mm long) counted on three consecutive days (-3, -2 and -1). To evaluate the therapeutic and residual efficacies of these formulations, tick counts (females ranging from 4.5 to 8.0 mm long) were performed on days 3,7 and 14 post-treatment, and continued on a weekly basis thereafter until the end of each experiment. The results obtained throughout this study, utilizing field efficacy studies, allowed us to conclude that the resistance of R. (B.) microplus against 200 and 500 mu g/kg ivermectin is widely disseminated because all tick populations that had contact with these specific concentrations were diagnosed as resistant. However, it is possible to infer that R. (B.) microplus resistance against 630 mu g/kg ivermectin was also widespread, diagnosed at six of ten analyzed properties. Resistance of these ectoparasites to 630 mu g/kg ivermectin is most likely emerging in three other populations of R. (B.) microplus. Strategies of resistance management need to be quickly determined to keep the selection pressure at a minimum level in Brazil. (C) 2014 Elsevier B.V. All rights reserved.

Differentiation and species status of the Neotropical yellow-eared bats Vampyressa pusilla and V. thyone (Phyllostomidae) with a molecular phylogeny and review of the genus

A systematic re-evaluation of Vampyressa pusilla warrants the elevation of V. p. thyone from subspecies to species rank based on its distinction from the allopatric V. p. pusilla. Morphological, mensural, chromosomal, and mitochondrial differences define each of these two taxa as divergent lineages. Vampyressa pusilla is endemic to the Atlantic Forest of southeastern South America and V. thyone is found allopatrically in northwestern South America, Central America, and southern Mexico. A molecular phylogenetic analysis of the mtDNA ND3-4 gene region using restriction endonuclease cut sites resulted in a monophyletic, although weakly supported Vampyressa ingroup with Chiroderma, and a clade of Mesophylla and Ectophylla as successive basal outgroup lineages. The phylogeny within Vampyressa, with the exception of V. melissa which is most similar to V. thyone based on karyotypes and morphology, had a topology of ((pusilla + thyone) + ((brocki + nymphaea) + bidens))).

Icnofósseis (paleotocas e crotovinas) atribuídos a mamíferos extintos no sudeste e sul do Brasil

ICHNOFOSSILS (PALEO-BURROWS and CROTOVINES) ATTRIBUTED TO EXTINCT MAMMALS IN SOUTHEASTERN and SOUTH BRAZIL. This work presents information regarding tunnels which are attributed to large extinct mammals. These structures can be found in several places in southeastern and southern Brazil, in different types of substrate, occurring as hollow structures (paleo-burrows) or those filled with sediments (crotovines). The dimensions and osteoderm and claw imprints found along the internal walls of the paleo-burrow found on aluvial fan deposits near the town of Cristal (Rio Grande do Sul State) suggest that a dasypodid xenarthran might have dug this structure. Comparison with similar structures found in Argentina can provide more detailed information regarding the paleoecology and biostratigraphy of the organisms that made these burrows.

Polymorphism analysis of the hsp70 stress gene in Broiler chickens (Gallus gallus) of different breeds

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

Estrutura e composição da ictiofauna de riachos do rio Paranapanema, Sudeste e Sul do Brasil

Foram amostrados 17 trechos de riachos com 100 m de extensão, todos de ordem igual ou menor a três, ao longo de ambas as margens do canal principal do Rio Paranapanema, SP e PR. O ponto médio de cada trecho foi georreferenciado via satélite com receptor GPS e o uso de metodologia padronizada de coleta de dados ambientais e peixes (baseada principalmente na pesca elétrica), possibilitou a obtenção das seguintes informações em cada local: 1) composição taxonômica da ictiofauna e contribuição, em termos de número de indivíduos e biomassa, de cada espécie para a ictiofauna local como um todo; 2) documentação fotográfica de espécimes representativos de cada espécie coletada com sua coloração natural; 3) descrição de cada ambiente coletado, com ilustrações fotográficas coloridas, e seus principais parâmetros bióticos e abióticos. No total foram coletados 3.683 exemplares, pertencentes a seis ordens, 16 famílias, 37 gêneros e 52 espécies, com biomassa total de 16,8 kg. Das espécies coletadas, aproximadamente 36% pertencem a ordem Siluriformes, 36% a Characiformes, 11% a Gymnotiformes, 10% a Perciformes, 4% a Cyprinodontiformes e 2% a Synbranchiformes. As espécies mais abundantes em termos de número de indivíduos foram Astyanax altiparanae (15,2%) e Astyanax sp. 1 (12,3%); aquelas com maior biomassa foram A. altiparanae (28%) e Geophagus brasiliensis (13%). A composição da ictiocenose em termos de abundância e biomassa por família indica a predominância expressiva de Characidae, seguida por Loricariidae, Pimelodidae e Cichlidae. Dentre os trechos amostrados, o trecho 14 (24 espécies) e o 13 (cinco espécies), apresentaram a maior e a menor riqueza em espécies, respectivamente, coincidindo com os valores obtidos para o índice de diversidade específica de Shannon-Wienner (H´= 0,99 e 0,32, respectivamente). A riqueza média encontrada foi de 11 espécies por trecho de riacho. Na estimativa de riqueza por extrapolação para o conjunto total de riachos amostrados na bacia do rio Paranapanema, obtivemos um valor de 69 espécies (erro padrão igual a quatro) indicando ser necessário um esforço amostral adicional moderado para atingir a assíntota da curva. Das 52 espécies coletadas, oito (aproximadamente 15% do total) são seguramente novas, cinco (aproximadamente 10% do total) possuem status taxonômico ainda indefinido, enquanto outras três (aproximadamente 6% do total) são espécies introduzidas. Analisando a estrutura trófica e espacial da ictiocenose estudada, as 10 espécies numericamente dominantes nos riachos estudados dividem-se, em ordem decrescente de importância numérica, em quatro guildas: onívoros nectônicos; invertívoros bentônicos; perifitívoros; e onívoros bentônicos. Uma chave de identificação para todas as espécies de peixes coletadas durante este estudo é fornecida ao final deste trabalho.

Caracterização do gene da proteína capsidial do Grapevine virus A em videiras afetadas pela acanaladura do lenho de Kober no Estado de São Paulo

O presente trabalho caracteriza o gene codificador da proteína capsidial do isolado do Grapevine virus A (GVA) encontrado no Estado de São Paulo (GVA-SP). RNA total foi extraído de folhas e pecíolos de plantas de videira (Vitis spp.) da variedade 'Kober 5BB' e submetido a RT-PCR usando oligonucleotídeos desenhados para amplificar um fragmento entre as posições 6409 e 7175 do RNA do GVA (GenBank, acesso X75433). Foi obtido um fragmento de tamanho esperado (767 nt) que inclui o gene da proteína capsidial, codificando 198 aminoácidos. A seqüência do GVA-SP apresentou similaridade de nucleotídeos e aminoácidos de, respectivamente, 86-92,3% e 94,5-98% com isolados do GVA da Europa, África e Japão (Acessos X75433, AF441234, AF007415, AB039841) e da região Sul do Brasil (Acesso AF494187), sendo, entretanto, mais similar aos isolados africano e italiano.

Ecological aspects of helminth fauna of Magellanic penguins, Spheniscus magellanicus (aves: Spheniscidae), from the Northern Coast of the State of São Paulo, Brazil

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

Avaliação ultra-sonográfica do sistema urinário, figado e útero do macaco-prego, Cebus apella

O macaco-prego, Cebus apella, é muito difundido no norte e sul da Amazônia Legal Brasileira e no Cerrado. Estes animais encontram-se rotineiramente submetidos à caça predatória, aumentando assim a necessidade de preservação desta espécie silvestre. Realizou-se um estudo ultra-sonográfico de 10 macacos-prego como forma de descrever a anatomia ultra-sonográfica normal de sua cavidade abdominal. A vesícula urinária apresentou parede com espessura média 0,2cm e em posição anatômica cuja topografia permitiu contato com as paredes do corpo do útero e cólon descendente. À varredura abdominal caudal foi visualizada a aorta, veia cava caudal e veia ilíaca direita. O fígado foi visto em varredura sagital e transversal, possibilitando a observação da vesícula biliar e vasos hepáticos. A varredura renal demonstrou com precisão a pelve, seio renal e relação cortico-medular. O comprimento médio de ambos os rins foi de 6,24±0,31cm, não existindo diferença estatística entre o rim direito e esquerdo (Teste t de Student e ANOVA). O volume renal foi 2,37±0,18cm³. Os coeficientes de Correlação de Pearson entre os comprimentos renais direito e esquerdo e entre volumes renais direito e esquerdo foram dispostos como r = 0,74 e 0,51. As espessuras médias para a região cortical e medular foram 0,75±0,11cm e 0,39±0,06cm, respectivamente. O coeficiente de correlação para a relação cortico-medular entre os rins direito e esquerdo foi de r = 0,19. O exame ultrasonográfico mostrou-se como uma técnica eficiente, nãoinvasiva, rápida e reprodutível, que provê dados importantes aos profissionais da área de clínica e cirurgia de animais silvestres.

Multiple events of horizontal transfer of the Minos transposable element between Drosophila species

In this study the Minos element was analyzed in 26 species of the repleta group and seven species of the saltans group of the genus Drosophila. The PCR and Southern blot analysis showed a wide occurrence of the Minos transposable element among species of the repleta and the saltans groups and also a low number of insertions in both genomes. Three different analyses, nucleotide divergence, historical associations, and comparisons between substitution rates (d(N) and d(S)) of Minos and Adh host gene sequences, suggest the occurrence of horizontal transfer between repleta and saltans species. These data reinforce and extend the Arca and Savakis [Genetica 108 (2000) 263] results and suggest five events of horizontal transfer to explain the present Minos distribution: between D. saltans and the ancestor of the mulleri and the mojavensis clusters; between D. hydei and the ancestor of the mulleri and the mojavensis clusters; between D. mojavensis and D. aldrichi; between D. buzzatii and D. serido; and between D. spenceri and D. emarginata. An alternative explanation would be that repeated events of horizontal transfer involving D. hydei, which is a cosmopolitan species that diverged from the others repleta species as long as 14 Mya, could have spread Minos within the repleta group and to D. saltans. The data presented in this article support a model in which distribution of Minos transposon among Drosophila species is determined by horizontal transmission balanced by vertical inactivation and extinction. (c) 2004 Elsevier B.V. All rights reserved.

3 CHAMAEZA ANTTHRUSHES IN EASTERN BRAZIL (FORMICARIIDAE)

There are three, not two, sibling antthrushes of the genus Chamaeza (Formicariidae) in eastern Brazil. One is the short-tailed, large, pale-billed, lower montane C. campanisona, with a spotted throat, dark forehead, and a long song ending in several grunts. The second is the long-tailed, small, dark-billed, montane and southern C. ruficauda, with barred undertail coverts and a short upscale song. The third is medium-tailed, small, dark-billed, midmontane, with a long upscale song like that of midmontane so-called ruficauda in Colombia and Venezuela. This northern group is like the third Brazilian species in proportions but not in coloration, so is considered to be the separate species C. turdia. The third Brazilian bird is probably C. meruloides Vigors 1825, based on an 1826 color plate; type specimens were sold at auction and have disappeared. It has a reddish crown and olive-brown back as in C. campanisona but reddish forehead and tail as in C. ruficauda; the throat is unspotted but the rest of the underparts are as in C. campanisona. C. meruloides and C. turdina form a vocally similar superspecies.

Phylogeography of the Solanaceae-infecting Basidiomycota fungus Rhizoctonia solani AG-3 based on sequence analysis of two nuclear DNA loci

Background: the soil fungus Rhizoctonia solani anastomosis group 3 (AG-3) is an important pathogen of cultivated plants in the family Solanaceae. Isolates of R. solani AG-3 are taxonomically related based on the composition of cellular fatty acids, phylogenetic analysis of nuclear ribosomal DNA (rDNA) and beta-tubulin gene sequences, and somatic hyphal interactions. Despite the close genetic relationship among isolates of R. solani AG-3, field populations from potato and tobacco exhibit comparative differences in their disease biology, dispersal ecology, host specialization, genetic diversity and population structure. However, little information is available on how field populations of R. solani AG-3 on potato and tobacco are shaped by population genetic processes. In this study, two field populations of R. solani AG-3 from potato in North Carolina (NC) and the Northern USA; and two field populations from tobacco in NC and Southern Brazil were examined using sequence analysis of two cloned regions of nuclear DNA (pP42F and pP89).Results: Populations of R. solani AG-3 from potato were genetically diverse with a high frequency of heterozygosity, while limited or no genetic diversity was observed within the highly homozygous tobacco populations from NC and Brazil. Except for one isolate (TBR24), all NC and Brazilian isolates from tobacco shared the same alleles. No alleles were shared between potato and tobacco populations of R. solani AG-3, indicating no gene flow between them. To infer historical events that influenced current geographical patterns observed for populations of R. solani AG-3 from potato, we performed an analysis of molecular variance (AMOVA) and a nested clade analysis (NCA). Population differentiation was detected for locus pP89 (Phi(ST) = 0.257, significant at P < 0.05) but not for locus pP42F (Phi(ST) = 0.034, not significant). Results based on NCA of the pP89 locus suggest that historical restricted gene flow is a plausible explanation for the geographical association of clades. Coalescent-based simulations of genealogical relationships between populations of R. solani AG-3 from potato and tobacco were used to estimate the amount and directionality of historical migration patterns in time, and the ages of mutations of populations. Low rates of historical movement of genes were observed between the potato and tobacco populations of R. solani AG-3.Conclusion: the two sisters populations of the basidiomycete fungus R. solani AG-3 from potato and tobacco represent two genetically distinct and historically divergent lineages that have probably evolved within the range of their particular related Solanaceae hosts as sympatric species.

A comparative floristic analysis of a cerrado area in Botucatu, State of São Paulo (Brazil)

Based on the results of a phytosociological survey in a ''cerrado'' area located within the municipality of Botucatu, state of São Paulo Brazil, a comparative analysis of the 58 sampled tree species' is here presented using the data of other available floristics works for the ''cerrados in the same state. With a binary matrix of presence/absence of shared common species with the studied plot, a clustering analysis was performed thus obtaining a dendrogram which groups northern and southern areas with the same vegetation in the state of São Paulo previously treated as floristicaly differentiated. These results are discussed under the assumption that Isolated areas of ''cerrados'' are to be taken as targets for immediate conservation.

Tissue distribution of a plasmid DNA encoding Hsp65 gene is dependent on the dose administered through intramuscular delivery

In order to assess a new strategy of DNA vaccine for a more complete understanding of its action in immune response, it is important to determine the in vivo biodistribution fate and antigen expression. In previous studies, our group focused on the prophylactic and therapeutic use of a plasmid DNA encoding the Mycobacterium leprae 65-kDa heat shock protein (Hsp65) and achieved an efficient immune response induction as well as protection against virulent M. tuberculosis challenge. In the present study, we examined in vivo tissue distribution of naked DNA-Hsp65 vaccine, the Hsp65 message, genome integration and methylation status of plasmid DNA. The DNA-Hsp65 was detectable in several tissue types, indicating that DNA-Hsp65 disseminates widely throughout the body. The biodistribution was dose-dependent. In contrast, RT-PCR detected the Hsp65 message for at least 15 days in muscle or liver tissue from immunized mice. We also analyzed the methylation status and integration of the injected plasmid DNA into the host cellular genome. The bacterial methylation pattern persisted for at least 6 months, indicating that the plasmid DNA-Hsp65 does not replicate in mammalian tissue, and Southern blot analysis showed that plasmid DNA was not integrated. These results have important implications for the use of DNA-Hsp65 vaccine in a clinical setting and open new perspectives for DNA vaccines and new considerations about the inoculation site and delivery system. © 2006 Coelho-Castelo et al; licensee BioMed Central Ltd.

Genetic vaccine for tuberculosis (pVAXhsp65) primes neonate mice for a strong immune response at the adult stage

Background: Vaccination of neonates is generally difficult due to the immaturity of the immune system and consequent higher susceptibility to tolerance induction. Genetic immunization has been described as an alternative to trigger a stronger immune response in neonates, including significant Th1 polarization. In this investigation we analysed the potential use of a genetic vaccine containing the heat shock protein (hsp65) from Mycobacterium leprae (pVAXhsp65) against tuberculosis (TB) in neonate mice. Aspects as antigen production, genomic integration and immunogenicity were evaluated. Methods: Hsp65 message and genomic integration were evaluated by RT-PCR and Southern blot, respectively. Immunogenicity of pVAXhsp65 alone or combined with BCG was analysed by specific induction of antibodies and cytokines, both quantified by ELISA. Results: This DNA vaccine was transcribed by muscular cells of neonate mice without integration into the cellular genome. Even though this vaccine was not strongly immunogenic when entirely administered (three doses) during early animal's life, it was not tolerogenic. In addition, pVAXhsp65 and BCG were equally able to prime newborn mice for a strong and mixed immune response (Th1 + Th2) to pVAXhsp65 boosters administered later, at the adult life. Conclusion: These results suggest that pVAXhsp65 can be safely used as a priming stimulus in neonate animals in prime-boost similar strategies to control TB. However, priming with BCG or pVAXhsp65, directed the ensuing immune response triggered by an heterologous or homologous booster, to a mixed Th1/Th2 pattern of response. Measures as introduction of IL-12 or GM-CSF genes in the vaccine construct or even IL-4 neutralization, are probably required to increase the priming towards Th1 polarization to ensure control of tuberculosis infection. © 2007 Pelizon et al; licensee BioMed Central Ltd.

Potential use of the veris apparent EC sensor to predict soil texture under the semi-arid conditions of central Arizona

The operational details of the apparent electrical conductivity (ECa) sensor manufactured by Veris Technologies have been extensively documented in literature reports, but the geographical distribution of these research studies indicate a strong regional concentration in the US Mid-west and Southern states. The agricultural lands of these states diverge significantly to the soil conditions and water regime of irrigated land in the US South-western states such as Arizona where there is no previous research reports of the use of this particular sensor. The objectives of the present study were to analyze the performance of this sensor under the conditions of typical soils in irrigated farms of Central Arizona. We tested under static conditions the performance of the sensor on three soils of contrasting texture. Observations were collected as time series data as soil moisture changed from saturation to permanent wilting point. Observations were repeated at the hours of lowest and highest temperatures. In addition, this study included soil penetration resistance and salinity determinations. Preliminary results indicate that soil temperature of the upper layer caused the most dynamic change in the sensor output. The ECa curves of the three soil textures tested had well defined distinctive characteristics. Final multivariate analysis is pending.