Giants' cooperation: a draft genome of the giant ciliate Muniziella cunhai suggests its ecological role in the capybara's digestive metabolism.

Several hundred ciliate species live in animals' guts as a part of their microbiome. Among them, Muniziella cunhai (Trichostomatia, Pycnotrichidae), the largest described ciliate, is found exclusively associated with Hydrochoerus hydrochaeris (capybara), the largest known rodent reaching up to 90 kg. Here, we present the sequence, structural and functional annotation of this giant microeukaryote macronuclear genome and discuss its phylogenetic placement. The 85 Mb genome is highly AT rich (GC content 25.71 %) and encodes a total of 11 397 protein-coding genes, of which 2793 could have their functions predicted with automated functional assignments. Functional annotation showed that M. cunhai can digest recalcitrant structural carbohydrates, non-structural carbohydrates, and microbial cell walls, suggesting a role in diet metabolization and in microbial population control in the capybara's intestine. Moreover, the phylogenetic placement of M. cunhai provides insights on the origins of gigantism in the subclass Trichostomatia.

Genetic structure of tree and shrubby species among anthropogenic edges, natural edges, and interior of an atlantic forest fragment.

Two species, Psychotria tenuinervis (shrub, Rubiaceae) and Guarea guidonia (tree, Meliaceae), were used as models to compare the genetic structure of tree and shrubby species among natural edges, anthropogenic edges, and a fragment interior. There were significant differences between two genetic markers. For isozymes, P. tenuinervis presented greater heterozygosity (expected and observed) and a higher percentage of polymorphic loci and median number of alleles than G. guidonia. For microsatellites, there was no difference in genetic variability between the species. Only P. tenuinervis, for isozymes, showed differences in genetic variability among the three habitats. There was no genetic structure (F (ST) < 0.05) among habitats in both plant species for both genetic markers. Isozymes showed great endogamy for both plant species, but not microsatellites. The forest fragmentation may have negative effects on both spatial (among edges and interior) and temporal genetic variability.

Alcohol dehydrogenase activities and ethanol tolerance in Anastrepha (Diptera, Tephritidae) fruit-fly species and their hybrids.

The ADH (alcohol dehydrogenase) system is one of the earliest known models of molecular evolution, and is still the most studied in Drosophila. Herein, we studied this model in the genus Anastrepha (Diptera, Tephritidae). Due to the remarkable advantages it presents, it is possible to cross species with different Adh genotypes and with different phenotype traits related to ethanol tolerance. The two species studied here each have a different number of Adh gene copies, whereby crosses generate polymorphisms in gene number and in composition of the genetic background. We measured certain traits related to ethanol metabolism and tolerance. ADH specific enzyme activity presented gene by environment interactions, and the larval protein content showed an additive pattern of inheritance, whilst ADH enzyme activity per larva presented a complex behavior that may be explained by epistatic effects. Regression models suggest that there are heritable factors acting on ethanol tolerance, which may be related to enzymatic activity of the ADHs and to larval mass, although a pronounced environmental effect on ethanol tolerance was also observed. By using these data, we speculated on the mechanisms of ethanol tolerance and its inheritance as well as of associated traits.

Gene flow and Andean uplift shape the diversification of Gasteracantha cancriformis (Araneae: Araneidae) in Northern South America.

The Andean uplift has played a major role in shaping the current Neotropical biodiversity. However, in arthropods other than butterflies, little is known about how this geographic barrier has impacted species historical diversification. Here, we examined the phylogeography of the widespread color polymorphic spider Gasteracantha cancriformis to evaluate the effect of the northern Andean uplift on its divergence and assess whether its diversification occurred in the presence of gene flow. We inferred phylogenetic relationships and divergence times in G. cancriformis using mitochondrial and nuclear data from 105 individuals in northern South America. Genetic diversity, divergence, and population structure were quantified. We also compared multiple demographic scenarios for this species using a model-based approach (phrapl) to determine divergence with or without gene flow. At last, we evaluated the association between genetic variation and color polymorphism. Both nuclear and mitochondrial data supported two well-differentiated clades, which correspond to populations occurring on opposite sides of the Eastern cordillera of the Colombian Andes. The final uplift of this cordillera was identified as the most likely force that shaped the diversification of G. cancriformis in northern South America, resulting in a cis- and trans-Andean phylogeographic structure for the species. We also found shared genetic variation between the cis- and trans-Andean clades, which is better explained by a scenario of historical divergence in the face of gene flow. This has been likely facilitated by the presence of low-elevation passes across the Eastern Colombian cordillera. Our work constitutes the first example in which the Andean uplift coupled with gene flow influenced the evolutionary history of an arachnid lineage.

Trypanosomatids (Protozoa: Kinetoplastida) in three species of Armored Catfish from Mogi-Guaçu river, Pirassununga, São Paulo, Brazil.

Trypanosome infections have been reported in several species of fish, in majority of cases described on the basis of morphological characteristics. Trypanosomes in fish are heteroxenous and transmitted by hirudineans. This study aims to evaluate the prevalence and density of infections by Trypanosoma sp. in blood from three species of catfish, Hypostomus regani, H. strigaticeps, H. albopunctatus, from the Mogi Guaçu River, Pirassununga, São Paulo, Brazil. Further, this study intends to characterize the Trypanosoma specimens found in the blood of these fish by morphological and molecular techniques. The trypanosomes overall prevalence observed was 47.6% with a general average density of 0.75 parasites/µl of blood. Hypostomus regani and Hypostomus strigaticeps showed a significant difference in prevalence. The average densities of parasites were not significantly different among the three fish species. Similar findings were observed for the monthly variations in densities. The parasites found in the three species of catfish studied showed similar morphological characteristics. The morphological data and the statistical analyses used in this study didn't show the formation of groups. The analyses provided evidence of the presence of pleomorphisms in the trypanosomes found in the three studied fish.

Trypanosomatids (Protozoa: Kinetoplastida) in three species of Armored Catfish from Mogi-Guaçu river, Pirassununga, São Paulo, Brazil / Tripanossomatídeos (Protozoa: Kinetoplastida) em três espécies de cascudos do rio Mogi-Guaçu, Pirassununga, São Paulo, Brasil

Abstract Trypanosome infections have been reported in several species of fish, in majority of cases described on the basis of morphological characteristics. Trypanosomes in fish are heteroxenous and transmitted by hirudineans. This study aims to evaluate the prevalence and density of infections by Trypanosoma sp. in blood from three species of catfish, Hypostomus regani, H. strigaticeps, H. albopunctatus, from the Mogi Guaçu River, Pirassununga, São Paulo, Brazil. Further, this study intends to characterize the Trypanosoma specimens found in the blood of these fish by morphological and molecular techniques. The trypanosomes overall prevalence observed was 47.6% with a general average density of 0.75 parasites/µl of blood. Hypostomus regani and Hypostomus strigaticeps showed a significant difference in prevalence. The average densities of parasites were not significantly different among the three fish species. Similar findings were observed for the monthly variations in densities. The parasites found in the three species of catfish studied showed similar morphological characteristics. The morphological data and the statistical analyses used in this study didn’t show the formation of groups. The analyses provided evidence of the presence of pleomorphisms in the trypanosomes found in the three studied fish.

Resumo Infecções por tripanossomas foram descritas em diversas espécies de peixes, sendo a maioria das descrições baseada nas características morfológicas. Tripanossomas de peixes são heteroxenos e transmitidos por hirudíneos. Este estudo tem como objetivo a avaliação da prevalência e densidade da infecção por Trypanosoma sp. no sangue de três espécies de cascudos, Hypostomus regani, H.strigaticeps, H.albopunctatus, do Rio Mogi-Guaçu, Pirassununga, São Paulo, Brasil. Além disso, pretende-se a caracterização das espécies de Trypanosoma encontradas no sangue desses peixes, por meio da utilização de técnicas morfólogicas e moleculares. A prevalência total de tripanossomas foi de 47,6%, e a densidade média foi de 0,75 parasitas/ µl de sangue. Hypostomus regani e Hypostomus strigaticeps apresentaram diferenças significativas em prevalência. As médias das densidades dos parasitas não apresentaram diferenças significativas entre as três espécies de peixes estudadas. O mesmo foi observado com a variação mensal das densidades. Os parasitas encontrados nas três espécies de cascudo possuíam características morfológicas semelhantes. As análises morfológicas e estatísticas obtidas neste estudo não mostraram a formação de grupos. As análises evidenciaram a presença de pleomorfismo dos tripanossomas encontrados nas três espécies de peixes estudados.

Mating systems of Psychotria tenuinervis (Rubiaceae): distance from anthropogenic and natural edges of Atlantic forest fragment.

The aim of this study was to determine and compare the mating systems among Psychotria tenuinervis populations at anthropogenic edges, natural edges, and the forest interior using allozyme electrophoresis of naturally pollinated progeny arrays. P. tenuinervis showed low outcrossing rates, varying from 37% to 50% of the mating attributable to outcrossing and 50% to 63% attributable to self-fertilization, in the three habitats. The forest interior had the highest outcrossing rate (t(m) = 0.50 and t(s) = 0.43) among the three habitats. However, there were no differences in either multilocus or single-locus rates among the three habitats, indicating that the contribution of biparental inbreeding to the apparent selfing rate in these populations was very low. The multilocus (t(m)) and single-locus (t(s)) outcrossing rates for the P. tenuinervis in the sample plots within each habitat showed great heterogeneity. In conclusion, edge creation seems not to influence its mating systems. Additionally, although P. tenuinervis is a distylous species, the population's inbreeding can be attributed almost entirely to self-fertilization.

Genetic variation and population structuring in two brooding coral species (Siderastrea stellata and Siderastrea radians) from Brazil.

Siderastrea stellata and S. radians are scleractinian coral species that present a remarkable overlap of diagnostic characteristics and sympatric distribution. Moreover, both are viviparous with similar reproductive strategies and with a gregarious larval behavior. Samples of both species from the Brazilian coast were analyzed using 18 isozymic loci to quantify their genetic variability and populational structure. Results confirmed species identity, high intrapopulational variability and revealed moderate genetic structuring among all samples (S. stellata: F (ST) = 0.070; S. radians: F (ST) = 0.092). Based on genotypic diversity analysis, there was evidence that local recruitment may have a minor role in the populations (mean, G ( o ):G ( e )= 1.00 +/- 0.0003 SD for S. stellata and 0.99 +/- 0.0023 SD for S. radians). Deviations towards heterozygote deficiencies found in both Siderastrea species could be explained by the Wahlund effect, since there was evidence that populations might be composed of colonies of different ages. In S. radians it is also likely that there is some inbreeding occurring in the studied populations. Despite the brooding pattern and the gregarious larval behavior, our data suggest the occurrence of gene flow along the Brazilian coast. This is the first study on population genetics of Brazilian reef corals.

Population genetics of Collisella subrugosa (Patellogastropoda: Acmaeidae): evidence of two scales of population structure.

Marine invertebrate populations usually show high levels of genetic variability that has frequently been associated with spatial and temporal environmental heterogeneity. One of the most heterogeneous marine environments is the intertidal zone, the habitat of Collisella subrugosa, the most widespread and abundant Brazilian limpet. C. subrugosa has planktonic larvae that can disperse over long distances, what can promote gene flow among shores, working against interpopulational differentiation. In this study we investigated the genetic variability and populational substructure of C. subrugosa through analysis of 24 allozyme loci in 14 samples (590 individuals) collected along 2,700 km of the Brazilian coast. The genetic variability was high ([Formula: see text] and [Formula: see text]), as expected for intertidal species. Genetic differentiation among samples was low (F (ST) = 0.03) what may reflect intensive gene flow associated with larval dispersal. However, we detected an isolation-by-distance pattern of population substructure in one sampled region. High levels of heterozygote deficiency were also observed for many loci in each sample. Alternative hypothesis are discussed, and the "breeding groups" is suggested to explain these pattern, indicating the main cause as environmental heterogeneity.

Quality of seeds produced by Psychotria tenuinervis (Rubiaceae): distance from anthropogenic and natural edges of Atlantic forest fragment.

The aim of this study was to investigate whether there were differences in the genetic variability and rate and velocity of the seed germination produced by Psychotria tenuinervis located at anthropogenic edges, natural edges, and in the forest interior. The populations of P. tenuinervis showed no differences in genetic variability or structure among the three habitats. There was, however, an indication of inbreeding, which was significantly higher in natural edges than in anthropogenic edges and the forest interior. Within-habitat variation was considerable, but there were no differences in seed mass or rate and velocity of germination among the three habitats. These results suggest that seed characteristics were not influenced by the genetic pattern of P. tenuinervis and that other characteristics of the forest fragment, such as gaps, edge age, and type of matrix exert more influence on seed mass and germination than the distance from the edges.

Low genetic structure in an epiphytic Orchidaceae (Oncidium hookeri) in the Atlantic rainforest of South-eastern Brazil.

BACKGROUND AND AIMS: Oncidium hookeri is a neotropical species of epiphytic Orchidaceae found in the Brazilian Atlantic rainforest at the top of the Mantiqueira Range of mountains. The genetic variation of O. hookeri was studied to assess the distribution of genetic variability within and among six populations localized in Atlantic rainforest remnants. Gene flow among populations and the occurrence of recent bottlenecks were investigated in order to infer the degree of isolation of these populations. METHODS: Thirteen polymorphic loci were used for allozyme electrophoresis. The data were analysed by means of standard statistical approaches, to estimate gene diversity and the genetic structure of the populations. KEY RESULTS: The mean gene diversity and allelic richness were H(e) = 0.099 and A = 1.75, respectively. F-statistics revealed high heterozygote deficiencies in all populations (F(IS) = 0.43-0.82). Several rare alleles were found in all the populations, and three populations presented private alleles. Low genetic differentiation among O. hookeri populations was detected (F(ST) = 0.029); natural selection may be involved in PGM locus differentiation among populations. The genetic differentiation between paired populations was low, bearing no correlation with geographic distance (Mantel test: r = -0.34, P = 0.72). Only two populations showed signs of recent bottlenecks. CONCLUSIONS: The heterozygote deficiency found seems to be caused by pollinator behaviour; the low frequencies of several alleles of different loci can be maintained due to clonal propagation. Despite the stochastic nature of the wind-dispersal of seeds to long distances, this process may promote an effective gene flow among populations, thus avoiding genetic differentiation.

Low allozymic variation in the Bidens pilosa L. complex (Asteraceae).

Bidens alba, B. subalternans, and B. pilosa form a complex group based on their morphological similarities. Bidens pilosa L. and B. subalternans DC. are herbs with a wide distribution in agricultural and disturbed areas. Bidens alba (L.) DC. varies in size from herb to subshrub and has a coastal distribution. Enzyme electrophoresis was used to evaluate genetic diversity in 12 populations of Bidens. All but three loci (Lap-1, Est, and Got) were monomorphic. Est-1 and Got were polymorphic only in B. alba. Lap-1 was polymorphic only in B. pilosa and B. subalternans. The estimates of genetic variability were low for all three taxa and all of the populations studied. Genetic diversity varied from 0.01 to 0.03. Mean genetic identities were high among populations of each species (0.99 for B. alba and 1.00 for B. pilosa and B. subalternans) and among the three species (1.00). Bidens pilosa and B. subalternans could be considered a single species if the taxonomy of the group were based solely on isozyme data.

Phylogenetic relationships of the New World Troidini swallowtails (Lepidoptera: Papilionidae) based on COI, COII, and EF-1alpha genes.

A phylogeny of the Neotropical members of the Tribe Troidini (Lepidoptera: Papilionidae) was obtained with sequences of three protein-coding genes: two mitochondrial (COI and COII), and one nuclear (EF-1alpha). Parsimony and Bayesian analyses of 33 taxa resulted in very similar trees regardless of method used with the 27 troidines always forming a monophyletic clade. Within Troidini, the genus Battus is sister group to the remaining troidines, followed by a clade formed by the Paleotropical taxa (here represented by three exemplars). The genus Euryades is the next branch, and sister group of Parides. The genus Parides is monophyletic, and is divided into four main groups by Maximum Parsimony analysis, with the most basal group composed of tailed species restricted to SE Brazil. Character optimization of ecological and morphological traits over the phylogeny proposed for troidines indicated that the use of several species of Aristolochia is ancestral over the use of few or a single host-plant. For the other three characters, the ancestral states were the absence of long tails, forest as the primary habitat and oviposition solitary or in loose group of several eggs.

Alcohol dehydrogenase activities and ethanol tolerance in Anastrepha (Diptera, Tephritidae) fruit-fly species and their hybrids

The ADH (alcohol dehydrogenase) system is one of the earliest known models of molecular evolution, and is still the most studied in Drosophila. Herein, we studied this model in the genus Anastrepha (Diptera, Tephritidae). Due to the remarkable advantages it presents, it is possible to cross species with different Adh genotypes and with different phenotype traits related to ethanol tolerance. The two species studied here each have a different number of Adh gene copies, whereby crosses generate polymorphisms in gene number and in composition of the genetic background. We measured certain traits related to ethanol metabolism and tolerance. ADH specific enzyme activity presented gene by environment interactions, and the larval protein content showed an additive pattern of inheritance, whilst ADH enzyme activity per larva presented a complex behavior that may be explained by epistatic effects. Regression models suggest that there are heritable factors acting on ethanol tolerance, which may be related to enzymatic activity of the ADHs and to larval mass, although a pronounced environmental effect on ethanol tolerance was also observed. By using these data, we speculated on the mechanisms of ethanol tolerance and its inheritance as well as of associated traits.

Ultra morphology of the digestive system of Anastrepha fraterculus AND Ceratitis capitata (DIPTERA TEPHRITIDAE)

Anastrepha fraterculus and Ceratitis capitata are widely distributed fruit flies that cause significant damageto fruit crops in tropical and temperate regions. The economic importance of these flies has resulted innumerous studies of their biology, with particular emphasis on their control and management. However,various aspects of the biology of these species are still poorly understood. In this work, we used scanningelectron microscopy (SEM) to examine the external anatomy and organization of the digestive system inthese two species. Adult males and females of A. fraterculus and females of C. capitata were dissected inphysiological saline solution, and the digestive tracts were removed and prepared for microscopy. SEMshowed that the crop was covered by a strong muscular layer that consisted of circular fibers connected bylongitudinal fibers; this arrangement was probably related to the post-feeding behavior of these flies in whichthe crop contents are regurgitated and reingested. The size of the rectum varied and was probably related tothe different body sizes of the two species.

Biological and genetic characteristics of uropathogenic Escherichia coli strains

The aim of the present study was to determine biological characteristics such as expression of fimbriae, Congo red binding, production of hemolysin and aerobactin, adhesion to HeLa and uroepithelial cells and invasion of HeLa cells by Escherichia coli isolates obtained from patients showing clinical signs of urinary tract infection (UTI). Also, the presence of genes (apa, afa, spa) for fimbria expression and cytotoxic necrotizing factors (CNF1, CNF2) was assayed using specific primers in PCR. The data obtained were compared with the clonal relationships obtained by analysis of multilocus enzyme electrophoresis (MLEE), restriction fragment length polymorphism (RFLP) of the rDNA (ribotyping) and enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR). All isolates but one presented a combination of at least two of the characteristics studied, a fact suggesting the presence of pathogenicity islands (PAIs). Diffuse adherence type to HeLa cells was observed to occur in most of the strains, but adhesion to uroepithelial cells seems to be a more reliable test to verify pathogenicity. Although four strains seemed to be able to invade HeLa cells when assayed by light microscopy, electron microscopy studies demonstrated that these strains were not invasive. MLEE, RFLP and ERIC-PCR were able to group the isolates differently into main clusters that were not correlated with the presence of pathogenic traits

Evolutionary trends of alcohol dehydrogenase isozymes in some species of tephritid flies

Com a finalidade de se caracterizar o padräo de evoluçäo das desidrogenases alcoolicas de tefritídeos foram realizados diversos tipos de análise. Através de eletroforese em gel de amido, foram analisadas as ADHs de larvas de alguns tefritídeos e de famílias relacionadas (drosofilídeos e otitídeos). Esta análise revelou uma grande variaçäo quanto ao número de locos que produzem este tipo de enzima. Em Tomoplagia rudolfi, näo se observou qualquer atividade de ADH. Em Acinia fucata, Rachiptera limbata, Rhagoletis nova e R. conversa observou-se apenas um loco, sendo que nas duas primeiras espécies, a enzima correspondente apresentou migraçäo anódica e atividade fortemente preferencial em alcoois de cadeia longa. Esta enzima corresponde ao loco Adh-2 e apresenta comportamento semelhante ao da enzima produzida pelo loco Odh de Drosophila melanogaster. Nas duas espécies estudadas do gênero Rhagoletis, a enzima produzida pelo loco que foi denominado Adh-1 apresentou migraçäo catódica e menor especialidade por substratos alcoólicos, mas ativa principalmente em alcoois de cadeia curta, similar ao observado nas enzimas produzidas pelo loco Adh de Drosophila melanogaster. As larvas de Euxesta eluta (Otitidae)) apresentaram uma ADH que se comportava de modo similar àquela produzida pelas larvas das espécies estudadas de Rhagoletis. Em Ceratitis capitata, Anastrepha grandis, A. fraterculus e A. striata foram observados dois locos, com as características dos locos Adh-1 e Adh-2 já comentadas, mas com especificidade de substrato bem menos pronunciada. Em A. obliqua, A. serpentina e A. bistrigata, foram observados, além destes dois locos, um terceiro (Adh-3), produzindo enzimas com características de migraçäo e especificidade de substrato semelhantes às das enzimas produzidas pelo loco Adh-2. Em A. bistrigata, foi detectado, em parte dos indivíduos o que seria uma enzima correspondente a um quarto loco, com características semelhantes às da enzima produzida pelo loco Odh de D. melanogaster. Medidas de atividade específica de ADH e do conteudo de etanol (principal alcool encontrado) nos frutos hospedeiros de espécies de Anastrepha mostraram haver uma correlaçäo positiva e estatisticamente signioficante entre estes parâmetros