Molecular phylogeny of land and freshwater planarians (Tricladida, Platyhelminthes): from freshwater to land and back

The suborder Tricladida (phylum Platyhelminthes) comprises the well-known free-living flatworms, taxonomically grouped into three infraorders according to their ecology: Maricola (marine planarians), Paludicola (freshwater planarians), and Terricola (land planarians). Molecular analyses have demonstrated that the Paludicola are paraphyletic, the Terricola being the sister group of one of the three paludicolan families, the Dugesiidae. However, neither 18S rDNA nor COI based trees have been able to resolve the relationships among species of Terricola and Dugesiidae, particularly the monophyly of Terricola. Here, we present new molecular data including sequences of nuclear genes (18S rDNA, 28S rDNA) and a mitochondrial gene (COI) of a wider sample of dugesiid and terricolan species. The new sequences have been analyzed, together with those previously obtained, in independent and concatenated analyses using maximum likelihood and bayesian methods. The results show that, although some parts of the trees remain poorly resolved, they support a monophyletic origin for Terricola followed by a likely return of some species to freshwater habitats. Relationships within the monophyletic group of Dugesiidae are clearly resolved, and relationships among some terricolan subfamilies are also clearly established and point to the need for a thorough revision of Terricola taxonomy.     

Global diversity of land planarians (Platyhelminthes, Tricladida, Terricola): a new indicator-taxon in biodiversity and conservation studies

Biodiversity conservation requires prioritization of areas for in situ conservation. In that perspective, the present study documents the global diversity of a component of the soil macrofauna, the land planarians, and concerns an exploratory analysis of their possible role as indicators of biodiversity. Diversity is described by three quantitative methods: (1) hotspots of species richness, selecting areas richest in species, (2) hotspots of range-size rarity, identifying areas richest in narrowly endemic species, and (3) complementarity, prioritizing areas according to their greatest combined species richness. The biodiversity measures of species richness and range-size rarity show a great correspondence in the identification of hotspots of diversity; both measures identify the following seven areas as the most biodiverse for land planarians: Sao Paulo, Florianopolis, western Java, Tasmania, Sri Lanka, North Island/New Zealand, and Sydney. It is discussed to what extent the results for the land planarians correspond with those obtained in other studies that assessed biodiversity hotspots for taxa on a global scale. It is noteworthy that land planarians identify a few global hotspots of diversity that generally do not feature, or only have low rankings, in other studies: New Zealand, southeastern Australia, and Tasmania.     

Biogeography and karyotypes of freshwater planarians (Platyhelminthes, Tricladida, Paludicola) in southern Brazil

In the Tricladida (Platyhelminthes), the incidence of different biotypes identified by several ploidy levels is very common. Planarians collected in the State of Rio Grande do Sul were identified using cytogenetics. Different species distributions were observed with respect to Rio Grande do Sul's geomorphology, which could have been caused by their different microhabitats. Girardia tigrina and G. anderlani consisted of diploid and triploid individuals, whereas G. schubarti showed diploids, triploids, and mixoploids; for all these species, individuals of different ploidies were sympatric. Only for diploid G. anderlani were B chromosomes observed. These B chromosomes seem to have an irregular segregational behavior during mitosis, and possibly also during meiosis. However the processes (e.g., selection, mutation) of maintaining 2n, 3n, and 2n/3n individuals within natural populations of G. schubarti remain to be clarified.     

Repeated DNA elements in planarians of the Dugesia gonocephala group (Platyhelminthes,Tricladida)

The results of the isolation of repetitive DNA elements in the genome of Dugesia etrusca, a species of the Dugesia gonocephala group, are reported. These sequences, about 1.4 kb long, represent only a part of longer interspersed genomic units (De1 family) and appear to be limited to the genome of some planarians of this group, as indicated by a Southern blot analysis performed in different species and populations. The genomic relationships among different species and populations of the D. gonocephala group are discussed in relation to the results obtained in the present work. This revised version was published online in July 2006 with corrections to the Cover Date.     

Land planarians (Platyhelminthes) as a model organism for fine-scale phylogeographic studies: understanding patterns of biodiversity in the Brazilian Atlantic Forest hotspot

The Brazilian Atlantic Forest is one of the richest biodiversity hotspots of the world. Paleoclimatic models have predicted two large stability regions in its northern and central parts, whereas southern regions might have suffered strong instability during Pleistocene glaciations. Molecular phylogeographic and endemism studies show, nevertheless, contradictory results: although some results validate these predictions, other data suggest that paleoclimatic models fail to predict stable rainforest areas in the south. Most studies, however, have surveyed species with relatively high dispersal rates whereas taxa with lower dispersion capabilities should be better predictors of habitat stability. Here, we have used two land planarian species as model organisms to analyse the patterns and levels of nucleotide diversity on a locality within the Southern Atlantic Forest. We find that both species harbour high levels of genetic variability without exhibiting the molecular footprint of recent colonization or population expansions, suggesting a long-term stability scenario. The results reflect, therefore, that paleoclimatic models may fail to detect refugia in the Southern Atlantic Forest, and that model organisms with low dispersal capability can improve the resolution of these models.     

Occurrence and abundance of a mariner-like element in freshwater and terrestrial planarians (Platyhelminthes, Tricladida) from southern Brazil

Transposable elements are DNA sequences present in all the large phylogenetic groups, both capable of changing position within the genome and constituting a significant part of eukaryotic genomes. The mariner family of transposons is one of the few which occurs in a wide variety of taxonomic groups, including freshwater planarians. Nevertheless, so far only five planarian species have been reported to carry mariner-like elements (MLEs), although several different species have been investigated. Regarding the number of copies of MLEs, Girardia tigrina is the only planarian species in which this has been evaluated, with an estimation of 8,000 copies of the element per haploid genome. Preliminary results obtained in our laboratory demonstrated that MLE is found in a large number of different species of planarians, including terrestrial. With this in mind, the aim was to evaluate the occurrence and estimate the number of MLE copies in different planarian species collected in south Brazil. Twenty-eight individuals from 15 planarian species were analyzed. By using PCR and the hybridization of nucleic acids, it was found that MLE was present in all the analyzed species, the number of copies being high, probably over 10(3) per haploid genome.     

Investigation of the ultrastructure of Dendrocoelum constrictum (Platyhelminthes,Tricladida) spermatogenesis and mature spermatozoa

To add to our understanding of dendrocoelid spermatozoa and to describe additional phylogenetic characters, the ultrastructure of the testis was investigated in the subterranean freshwater planarian Dendrocoelum constrictum. This is the first study investigating spermatogenesis and spermatozoon ultrastructure in a subterranean freshwater planarian species. We found that the basic structure of spermatozoa in D. constrictum is similar to that of other Tricladida that have been studied previously. In fact, D. constrictum spermatozoa possess an elongated nucleus, one giant mitochondrion, and two subterminal flagella with a 9 + ‘1’ pattern. The flagella emerge together from one side of the spermatozoon. However, D. constrictum has some characteristics that have not yet been described for other freshwater planarians. In fact, the number of cortical microtubules reaches the maximum number in the anterior and middle part of region I, and then decrease until they disappear towards the posterior extremity of the spermatozoon. The extreme tip of the anterior region of the spermatozoon exhibits a specific external ornamentation of the plasma membrane.     

The effect of habitat fragmentation on the genetic structure of a top predator: loss of diversity and high differentiation among remnant populations of Atlantic Forest jaguars (Panthera onca)

Habitat fragmentation may disrupt original patterns of gene flow and lead to drift-induced differentiation among local population units. Top predators such as the jaguar may be particularly susceptible to this effect, given their low population densities, leading to small effective sizes in local fragments. On the other hand, the jaguar's high dispersal capabilities and relatively long generation time might counteract this process, slowing the effect of drift on local populations over the time frame of decades or centuries. In this study, we have addressed this issue by investigating the genetic structure of jaguars in a recently fragmented Atlantic Forest region, aiming to test whether loss of diversity and differentiation among local populations are detectable, and whether they can be attributed to the recent effect of drift. We used 13 microsatellite loci to characterize the genetic diversity present in four remnant populations, and observed marked differentiation among them, with evidence of recent allelic loss in local areas. Although some migrant and admixed individuals were identified, our results indicate that recent large-scale habitat removal and fragmentation among these areas has been sufficiently strong to promote differentiation induced by drift and loss of alleles at each site. Low estimated effective sizes supported the inference that genetic drift could have caused this effect within a short time frame. These results indicate that jaguars' ability to effectively disperse across the human-dominated landscapes that separate the fragments is currently very limited, and that each fragment contains a small, isolated population that is already suffering from the effects of genetic drift.     

A new variant of the neodermatan-type spermiogenesis in a parasitic 'turbellarian', Notentera ivanovi (Platyhelminthes) and the origin of the Neodermata

Spermiogenesis is characterized by fully incorporated (in the testes of mature worms) or partially free (submature worms) axonemes in spermatids. Formation of free flagella correlates with tight arrangement of cells in the testes and small size of the zone of differentiation and vice versa . In both cases the axonemes elongate within the growing shaft, so that the organization of the resulting spermatids is different only with regard to the distal end. In late spermatids, the nucleus occupies the proximal half, the two mitochondria and the axonemes directed distally lie in the distal half. After detachment of the spermatid, a migration of the nucleus takes place. In the resulting mature sperm, the proximal (anterior) half is occupied by the mitochondria and axonemes the basal bodies of which lie at the anterior end of the spermatozoon; the nucleus occupies the distal (posterior) half. Because of the distal orientation of the axonemes and a peculiar mode of the migration of the nucleus, the spermiogenesis of Notentera should be classified as a new variant of the type characteristic of the Neodermata (parasitic Platyhelminthes). Based on the analysis of the available morphological and other relevant data it is argued (i) that a high-ranked taxon, the Fecampiida, should be established within the Neoophora to include Notentera and the closely related Fecampiidae and (ii) that all the Platyhelminthes with neodermatan-type spermiogenesis form a monophyletic taxon, the Revertospermata, which includes the sister groups Fecampiida and Mediofusata.     

A new and aberrant species of Dugesia (Platyhelminthes,Tricladida, Dugesiidae) from Madagascar

In this paper we report a new species of Dugesia of the family Dugesiidae from Madagascar, representing the fourth species of freshwater planarian known from this global biodiversity hotspot. In some respects the new species is aberrant, when compared with its congeners, being characterized by a head with smoothly rounded auricles, a peculiar course of the oviducts, including the presence of a common posterior extension, and by the asymmetrical openings of the vasa deferentia at about halfway along the seminal vesicle. Further, it is characterized by a ventral course of the ejaculatory duct with a terminal opening, very long spermiducal vesicles and unstalked cocoons. Its diploid chromosome complement with 18 chromosomes represents an uncommon feature among fissiparous species of Dugesia.     

Genetic variation in remnant populations of Dalbergia nigra (Papilionoideae), an endangered tree from the Brazilian Atlantic Forest

BACKGROUND AND AIMS: Dalbergia nigra, known as Brazilian rosewood, is an endangered tree species restricted to the Brazilian Atlantic Forest and has been intensively logged for five centuries due to its high-quality wood. The objective of the present study was to assess the genetic variation and structure in adults and saplings of the species from a large reserve of the Atlantic Forest, the Rio Doce State Park, and from two small surrounding fragments, one better preserved and another with a high degree of anthropogenic disturbance. METHODS: Analyses of genetic variation and structure were conducted by studying allozyme markers. Seven putative enzymatic loci were resolved, five of them being polymorphic. KEY RESULTS: The mean numbers of alleles per locus (A) were 1.93 and 1.73, while the percentages of polymorphic loci (P) were 93 and 73 % for adults and saplings, respectively. Saplings from the fragment with high anthropogenic disturbance exhibited the lowest values of A and P. The fragment that constitutes a conservation area exhibited genetic variation similar to the population from the large reserve. The observed (H(o)) and expected (H(e)) heterozygosities were not significantly different among the three populations. Only sapling populations showed F(ST) values (divergence among populations) significantly different from zero over all studied loci. The fragment with high anthropogenic disturbance exhibited considerable genetic divergence in relation to the above-cited populations. CONCLUSIONS: The evaluated populations displayed mean levels of genetic variation intermediate to those expected for narrow and widespread species. The results suggest that fragments with similar area and geographical distance from a large protected reserve can exhibit different levels of genetic variation, depending on the degree of anthropogenic disturbance. The considerable genetic variation in the protected fragment points to the importance of adequate conservation of small fragments for the preservation of genetic variation in D. nigra.     

Ecological restoration, carbon sequestration and biodiversity conservation: The experience of the Society for Wildlife Research and Environmental Education (SPVS) in the Atlantic Rain Forest of Southern Brazil

Since 1999, SPVS has been involved in three projects that combine two fundamental goals over the course of 40 years: the conservation of one of Brazil's most important remnants of Atlantic Forest and the implementation of projects for carbon sequestration. In addition, there is an interest in replicating these projects in order to restore other degraded areas, protect the Brazilian biomes, and help to diminish deforestation and forest fire, therefore reducing carbon emissions. The acquisition of 19,000 ha of degraded areas of high biological importance in southern Brazil was the first step towards the implementation of the projects. These areas are owned by SPVS, a Brazilian NGO, and are being restored, conserved and transformed into Private Natural Reserves, in partnership with the NGO – The Nature Conservancy, and financed by the companies – American Electric Power, General Motors and Chevron Texaco. The process of forest restoration involves several stages: soil studies, surveying the region's native plants, planning for restoration by means of a Geographical Information System, production of seedlings, application of different techniques for planting (such as manual or mechanised planting with seedlings and stakes), and biomass and biodiversity monitoring. To guarantee the survival of the seedlings on the planted areas, during the first three years, there is a continuous and systematic maintenance programme including weeding of undergrowth, crowing and organic fertilisation. The three projects already planted around 500,000 seedlings of native species until September 2004, and aim to plant a further 300,000 until 2008.     

Three new cecidogenous species of Palaeomystella Fletcher (Lepidoptera,Momphidae) from the Brazilian Atlantic Rain Forest

Three new cecidogenous species of Palaeomystella Fletcher (Lepidoptera, Momphidae) from the Brazilian Atlantic Rain Forest are described. Larvae of P. fernandesi Moreira & Becker, sp. n., P. rosaemariae Moreira & Becker, sp. n. and P. tavaresi Becker & Moreira, sp. n. induce galls, respectively, on Tibouchina sellowiana (Cham.) Cogn., T. asperior (Cham.) Cogn. and T. fissinervia (Schrank & Mart. ex DC.) Cogn. (Melastomataceae). Adults, immature stages and galls are illustrated, and data on life history and a preliminary analysis of mitochondrial DNA sequences, including related species, are also provided.     

Molecular barcoding and phylogeography of sexual and asexual freshwater planarians of the genus Dugesia in the Western Mediterranean (Platyhelminthes, Tricladida, Dugesiidae)

Planarians of the genus Dugesia have a worldwide distribution with high species diversity in the Mediterranean area. In this area, populations with a triploid karyotype that reproduce by fissiparity are exceptionally frequent, outnumbering the sexual populations. This situation poses interesting questions, such as the age of these asexual lineages, whether they all belong to the same species or whether the triploidization event is recurrent, and what factors (climatic, geographical, historical...) explain the prevalence of these asexual forms. However, asexual populations cannot be assigned to a species due to the lack of copulatory apparatus – the main structure used in species identification. In this study, we have developed a DNA barcoding method, based on COI and ITS-1 sequences, which allows the assignment of the fissiparous forms to sexual species. At the same time, phylogenetic analysis from species of the western Mediterranean have unveiled the presence of species with highly differentiated populations alongside species with a wide distribution and almost no genetic variation. The roles of habitat instability, dispersal capacity and human activities are briefly discussed.     

Morphogenia: a new genus of the Neotropical tribe Jubini (Coleoptera,Staphylinidae, Pselaphinae) from the Brazilian Amazon

A new genus and species of the large Neotropical pselaphine tribe Jubini is described from Manaus, Brazil, based on material preserved in the Natural History Museum, London. Morphogenia struhligen. et sp. n. represents the possible sister taxon of the abundant and speciose genus Barrojuba Park, sharing with it the putatively derived condition of anterolaterally shifted vertexal foveae, producing a smoothly convex vertex devoid of fovea or sulci. However, unlike Barrojuba, Morphogenia retains a plesiomorphic antebasal sulcus on the pronotum in both sexes, and additionally lacks elaborate abdominal fovea-like pockets and teeth on the lateral margins of the pronotum that are typical of Barrojuba. The genus is also unusual among jubine genera in lacking the characteristic V- or Y-shaped gular carina. In contrast to the commonly-collected Barrojuba, specimens of Morphogenia are absent in extensive jubine collections housed in museums in the United States, indicating that the new taxon may be relatively scarce or localised.     

Inversion polymorphism and a new polytene chromosome map of <Emphasis Type=Italic>Zaprionus indianus</Emphasis> Gupta (1970) (Diptera: drosophilidae)

Zaprionus indianus is a recent invader in Brazil and was probably introduced from the West Afrotropical zone. So far, studies regarding its chromosomal polymorphism were limited to India. We found that Brazilian populations were very different from Indian ones. Five new inversions have been discovered. In(II)A, already described in India, where it is quite common, has also been found in Brazil, where it is very rare. The X-chromosome has three inversions; In(X)Na, In(X)Ke and In(X)Eg, which are frequent in all Brazilian populations studied. In every case, we observed strong linkage disequilibrium among these gene arrangements. During the primary collection period (2001–2002), we noticed a significant positive correlation between the frequency of these inversions and latitude, but this was not confirmed in later investigations. Rearrangement In(IV)EF was also common in all populations, while inversion In(V)B was only found in southern populations. Our data suggest that the founders that recently invaded Brazil were polymorphic for the six inversions observed. The place of origin might be identified more precisely by investigating West African populations. In order to facilitate further investigations, we present an updated polytene chromosome photomap, locating the breakpoints of every inversion observed in Brazilian populations. Galina Ananina and Cláudia Rohde contributed equally to this work     

Behavioral activities and diet of Azaras’s capuchin monkey,Sapajus cay (Illiger, 1815), in a forest remnant of the Brazilian Cerrado

ABSTRACT

Azaras’s capuchin monkey (Sapajus cay) is a poorly studied species in ecological aspects. We investigated behavioral activities and diet of a group of S. cay in a remnant of the Brazilian Cerrado. We sought to identify the most frequent behavior that the group engaged in, as well as to verify possible differences in behavioral patterns and diet between adult males and females. From January to July 2012 we collected behavioral and foraging data for a group of 21 individuals using scan sampling. Behaviors and food items consumed were analyzed using percentages and two proportions Z-tests to assess differences between males and females. We obtained 878 scans in approximately 202 hours of sampling effort, resulting in 4,159 individual activity records of capuchin monkeys. The group allocated time to traveling (41.3%), resting (25.5%), feeding (14.3%), foraging (13.7%), and social activities (5.1%). Females spent more time foraging and feeding, while males spent more time resting. Fruits (61.4%), invertebrates (15.3%), and seeds (14.3%) were the most common food items consumed. We did not find differences in food consumption between sexes. Behavioral activities may be influenced by habitat quality of the studied area, opportunistic habits, and plasticity in the diet.     

Revision of the rhinoceros beetle genus Oryctophileurus Kolbe with description of a new species,the male of O. varicosus Prell,and notes on biogeography (Scarabaeoidea,Dynastinae, Phileurini)

The genus Oryctophileurus is reviewed and its validity is supported by a combination of the following apomorphic characters: a single cephalic horn with lateral carina, pronotal cavity with ocellate punctures and two teeth or tubercles close behind the anterior pronotal margin. The male of Oryctophileurus varicosus Prell, 1934, is described for the first time. A new species, Oryctophileurus guerrai Perger & Grossi sp. n., from subhumid Tucuman-Bolivian forest in the Southern Bolivian Andes is described. The new species is distinguished from its closest relative, O. armicollis Prell, 1911, by a narrower distance between the inner teeth of the dorsal pronotal protuberances and a reduced area of weakly developed ocellate punctures above the posterolateral pronotal margin. The occurrence of Oryctophileurus species in areas of endemism along the eastern slope of the tropical Andes suggests that these populations represent biogeographic “relicts”, and the discovery of Oryctophileurus guerrai sp. n. in the southern Bolivian Andes suggests that this area is underrated with respect to insect diversity and endemism.