Trypanosoma cruzi mitochondrial maxicircles display species- and strain-specific variation and a conserved element in the non-coding region

Abstract Background The mitochondrial DNA of kinetoplastid flagellates is distinctive in the eukaryotic world due to its massive size, complex form and large sequence content. Comprised of catenated maxicircles that contain rRNA and protein-coding genes and thousands of heterogeneous minicircles encoding small guide RNAs, the kinetoplast network has evolved along with an extreme form of mRNA processing in the form of uridine insertion and deletion RNA editing. Many maxicircle-encoded mRNAs cannot be translated without this post-transcriptional sequence modification. Results We present the complete sequence and annotation of the Trypanosoma cruzi maxicircles for the CL Brener and Esmeraldo strains. Gene order is syntenic with Trypanosoma brucei and Leishmania tarentolae maxicircles. The non-coding components have strain-specific repetitive regions and a variable region that is unique for each strain with the exception of a conserved sequence element that may serve as an origin of replication, but shows no sequence identity with L. tarentolae or T. brucei. Alternative assemblies of the variable region demonstrate intra-strain heterogeneity of the maxicircle population. The extent of mRNA editing required for particular genes approximates that seen in T. brucei. Extensively edited genes were more divergent among the genera than non-edited and rRNA genes. Esmeraldo contains a unique 236-bp deletion that removes the 5'-ends of ND4 and CR4 and the intergenic region. Esmeraldo shows additional insertions and deletions outside of areas edited in other species in ND5, MURF1, and MURF2, while CL Brener has a distinct insertion in MURF2. Conclusion The CL Brener and Esmeraldo maxicircles represent two of three previously defined maxicircle clades and promise utility as taxonomic markers. Restoration of the disrupted reading frames might be accomplished by strain-specific RNA editing. Elements in the non-coding region may be important for replication, transcription, and anchoring of the maxicircle within the kinetoplast network.

Phylogeography of the neotropical Anopheles triannulatus complex (Diptera: Culicidae) supports deep structure and complex patterns

Abstract Background The molecular phylogenetic relationships and population structure of the species of the Anopheles triannulatus complex: Anopheles triannulatus s.s., Anopheles halophylus and the putative species Anopheles triannulatus C were investigated. Methods The mitochondrial COI gene, the nuclear white gene and rDNA ITS2 of samples that include the known geographic distribution of these taxa were analyzed. Phylogenetic analyses were performed using Bayesian inference, Maximum parsimony and Maximum likelihood approaches. Results Each data set analyzed septely yielded a different topology but none provided evidence for the seption of An. halophylus and An. triannulatus C, consistent with the hypothesis that the two are undergoing incipient speciation. The phylogenetic analyses of the white gene found three main clades, whereas the statistical parsimony network detected only a single metapopulation of Anopheles triannulatus s.l. Seven COI lineages were detected by phylogenetic and network analysis. In contrast, the network, but not the phylogenetic analyses, strongly supported three ITS2 groups. Combined data analyses provided the best resolution of the trees, with two major clades, Amazonian (clade I) and trans-Andean + Amazon Delta (clade II). Clade I consists of multiple subclades: An. halophylus + An. triannulatus C; trans-Andean Venezuela; central Amazonia + central Bolivia; Atlantic coastal lowland; and Amazon delta. Clade II includes three subclades: Panama; cis-Andean Colombia; and cis-Venezuela. The Amazon delta specimens are in both clades, likely indicating local sympatry. Spatial and molecular variance analyses detected nine groups, corroborating some of subclades obtained in the combined data analysis. Conclusion Combination of the three molecular markers provided the best resolution for differentiation within An. triannulatus s.s. and An. halophylus and C. The latest two species seem to be very closely related and the analyses performed were not conclusive regarding species differentiation. Further studies including new molecular markers would be desirable to solve this species status question. Besides, results of the study indicate a trans-Andean origin for An. triannulatus s.l. The potential implications for malaria epidemiology remain to be investigated.

A new species of Leporinus Agassiz, 1829 from the upper Rio Paraná basin (Characiformes, Anostomidae) with redescription of L. elongatus Valenciennes, 1850 and L. obtusidens (Valenciennes, 1837)

Leporinus obtusidens Valenciennes, 1837 and L. elongatus Valenciennes, 1850 are redescribed based on the type specimens, including those of their junior synonyms, and recently collected specimens. Leporinus obtusidens is considered to be widespread, occuring in the river drainages of La Plata, São Francisco, and Parnaíba. Leporinus aguapeiensis Campos, 1945, described from the upper Rio Paraná, and L. silvestrii Boulenger, 1902, described from the Rio Paraguay, are considered junior synonyms of L. obtusidens. Leporinus elongatus is endemic to the Rio Jequitinhonha and Rio Pardo, two eastern Brazilian river basins, and the locality cited for the lectotype, Rio São Fransico, likely to be erroneous. Leporinus crassilabris Borodin, 1929, and L. crassilabris breviceps Borodin, 1929, both described from the Rio Jequitinhonha, are considered junior synynoms of L. elongatus. A new species of Leporinus, endemic to the upper Rio Paraná, very similar and sometimes mistaken with L. obtusidens, is formally described. In addition, comments on Leporinus pachyurus Valenciennes, 1850 and on L. bimaculatus Castelnau, 1855 are provided, and a lectotype for L. bimaculatus is selected.

Comparative analysis of the corps en cerise in several species of Laurencia (Ceramiales, Rhodophyta) from the Atlantic Ocean

Different species of Laurencia have proven to be a rich source of natural products yielding interesting bioactive halogenated secondary metabolites, such as terpenoids and acetogenins. It is shown that such compounds are accumulated in the spherical, reniform to claviform refractive inclusions called corps en cerise (CC), which are intensively osmiophilic and located mainly in the cortical cells of the thalli and also in trichoblast cells. Up to now, it was believed that CC were present only in these two kinds of cells. Recently, however, a species of Laurencia, L. marilzae, with CC in all cells of the thallus, i.e., cortical, medullary, including the pericentral and axial cells, as well as in the trichoblasts, was described from the Canary Islands, and subsequently also reported to Brazil and Mexico. Within the Laurencia complex, only Laurencia species produce CC. Since the species of Laurencia are targets of interest for the prospection of bioactive substances due to their potential antibacterial, antifungal, anticholinesterasic, antileishmanial, cytotoxic, and antioxidant activities, the present paper carries out a comparative analysis of the corps en cerise in several species of Laurencia from the Atlantic Ocean to obtain basic information that can support natural product bioprospection projects. Our results show that the number and size of the CC are constant within a species, independent of the geographical distribution, corroborating their use for taxonomical purposes to differentiate groups of species that present a lower number from those that have a higher number. In this regard, there was a tendency for the number of CC to be higher in some species of Laurencia from the Canary Islands. The presence of CC can also be used to distinguish species in which these organelles are present in all cells of the thallus from those in which CC are restricted to the cortical cells. Among the species analyzed, L. viridis displayed the most varied secondary metabolites composition, such as sesquiterpenes, diterpenes, triterpenes, all of which showed potent antiviral, cytotoxic, and antitumoral activities, including protein phosphatase type 2A (PP2A) inhibitory effects.

Comparison of primers for RAPD-PCR from environmental isolates of Cryptococcus neoformans, Cryptococcus albidus and Cryptococcus laurentii complex

Various organisms have been characterized by molecular methods, including fungi of the genus Cryptococcus. The purposes of this study were: to determine the discriminatory potential of the RAPD (Random Amplified Polymorphic DNA) primers, the pattern of similarity of the Cryptococcus species, and discuss their useful application in epidemiological studies. We analyzed 10 isolates of each specie/group: C. albidus, C. laurentii complex, C. neoformans var. grubii, all from environmental source, and two ATCC strains, C. neoformans var. grubii ATCC 90112, and C. neoformans var. neoformans ATCC 28957 by RAPD-PCR using the primers CAV1, CAV2, ZAP19, ZAP20, OPB11 and SEQ6. The primers showed a good discriminatory power, revealing important differences between them and between species; the SEQ6 primer discriminated a larger number of isolates of three species. Isolates of C. laurentii showed greater genetic diversity than other species revealed by all six primers. Isolates of C. neoformans were more homogeneous. Only the primer CAV2 showed no amplification of DNA bands for C. albidus. It was concluded that the use of limited number of carefully selected primers allowed the discrimination of different isolates, and some primers (e.g., CAV2 for C. albidus) may not to be applied to some species.

Changes in phytoplankton composition in response to tides, wind-induced mixing conditions, and freshwater outflows in an urbanised estuarine complex

Recent reports have shown an increase in potentially harmful phytoplankton in Santos bay (Southeastern Brazilian Coast), located in a highly urbanised estuarine complex. Prediction of blooms is, thus, essential but the phytoplankton community structure in very dynamic regions is difficult to determine. In the present work, we discriminate bloom forming microphytoplankton dominance and their relationship to physical and meteorological variables to look for patterns observed in different tides and seasons. Comparing 8 distinct situations, we found five scenarios of dominance that could be related to winds, tides and rainfall: i) Surfers, diatoms occurring during high surf zone energies; ii) Sinkers, represented by larger celled diatoms during spring tide, after periods of high precipitation rates; iii) Opportunistic mixers, composed of chain forming diatoms with small or elongate cells occurring during neap tides; iv) Local mixers, microplanktonic diatoms and dinoflagellates which occurred throughout the 298 sampling stations; and v) Mixotrophic dinoflagellates, after intense estuarine discharges. Results suggest alterations in the temporal patterns for some bloom-forming species, while others appeared in abundances above safe limits for public health. This approach can also illustrate possible impacts of changes in freshwater discharge in highly urbanised estuaries.

Fishes of Paranaguá Estuarine Complex, South West Atlantic

The objective of this work was to present an updated checklist of the currently known fishes in the Paranaguá Estuarine Complex (PEC) and provides comments on conservation status for the treated species. We used a large dataset derived from a pool of studies which have been conducted within there along the last 30 years. Each study was based on monthly samplings and conducted in several estuarine habitat; thus, the pool covers practically all estuarine habitats and takes into account the seasonal cycle in the system. The PEC ichthyofauna represents a mixture between that fauna typical from the tropical Brazilian coast and that with affinities of temperate Argentinean and Uruguayan zones. The PEC harbors a rich fish fauna of 213 species, inserted in the families that are common along the Brazilian coast. Only a minor part (8%) of the PEC fish fauna was evaluated as regards the conservation status, mostly because of the lack of basic biological and ecological information for most species. Despite part of the among-estuaries differences are due to different and incomplete sampling efforts, the richness in the PEC is surprisingly higher than other systems in Brazil and around world, which emphasize the importance of the region for global biodiversity conservation.