Diploid chromosome set of kissing bug Triatoma baratai (Hemiptera, Triatominae)

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

De novo transcriptome assembly for a non-model species, the blood-sucking bug Triatoma brasiliensis, a vector of Chagas disease

High throughput sequencing (HTS) provides new research opportunities for work on non-model organisms, such as differential expression studies between populations exposed to different environmental conditions. However, such transcriptomic studies first require the production of a reference assembly. The choice of sampling procedure, sequencing strategy and assembly workflow is crucial. To develop a reliable reference transcriptome for Triatoma brasiliensis, the major Chagas disease vector in Northeastern Brazil, different de novo assembly protocols were generated using various datasets and software. Both 454 and Illumina sequencing technologies were applied on RNA extracted from antennae and mouthparts from single or pooled individuals. The 454 library yielded 278 Mb. Fifteen Illumina libraries were constructed and yielded nearly 360 million RNA-seq single reads and 46 million RNA-seq paired-end reads for nearly 45 Gb. For the 454 reads, we used three assemblers, Newbler, CAP3 and/or MIRA and for the Illumina reads, the Trinity assembler. Ten assembly workflows were compared using these programs separately or in combination. To compare the assemblies obtained, quantitative and qualitative criteria were used, including contig length, N50, contig number and the percentage of chimeric contigs. Completeness of the assemblies was estimated using the CEGMA pipeline. The best assembly (57,657 contigs, completeness of 80 %, < 1 % chimeric contigs) was a hybrid assembly leading to recommend the use of (1) a single individual with large representation of biological tissues, (2) merging both long reads and short paired-end Illumina reads, (3) several assemblers in order to combine the specific advantages of each.

Interactions of Bacillus thuringiensis bioinsecticides and the predatory stink bug Podisus nigrispinus to control Plutella xylostella

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparing de novo and reference-based transcriptome assembly strategies by applying them to the blood-sucking bug Rhodnius prolixus

High Throughput Sequencing capabilities have made the process of assembling a transcriptome easier, whether or not there is a reference genome. But the quality of a transcriptome assembly must be good enough to capture the most comprehensive catalog of transcripts and their variations, and to carry out further experiments on transcriptomics. There is currently no consensus on which of the many sequencing technologies and assembly tools are the most effective. Many non-model organisms lack a reference genome to guide the transcriptome assembly. One question, therefore, is whether or not a reference-based genome assembly gives better results than de novo assembly. The blood-sucking insect Rhodnius prolixus-a vector for Chagas disease-has a reference genome. It is therefore a good model on which to compare reference-based and de novo transcriptome assemblies. In this study, we compared de novo and reference-based genome assembly strategies using three datasets (454, Illumina, 454 combined with Illumina) and various assembly software. We developed criteria to compare the resulting assemblies: the size distribution and number of transcripts, the proportion of potentially chimeric transcripts, how complete the assembly was (completeness evaluated both through CEGMA software and R. prolixus proteome fraction retrieved). Moreover, we looked for the presence of two chemosensory gene families (Odorant-Binding Proteins and Chemosensory Proteins) to validate the assembly quality. The reference-based assemblies after genome annotation were clearly better than those generated using de novo strategies alone. Reference-based strategies revealed new transcripts, including new isoforms unpredicted by automatic genome annotation. However, a combination of both de novo and reference-based strategies gave the best result, and allowed us to assemble fragmented transcripts.

Towards biological control strategies for the bronze bug, Thaumastocoris peregrinus, on eucalyptus plantations in South America

The bronze bug is an invasive Australian pest that has reached eucalypt production areas worldwide in <10 years. The fi rst record in South America was in 2005 in Argentina. Collaboration in the region towards a unifi ed strategy for the management of the bronze bug started soon after the dispersal of the pest into Brazil and Uruguay was confi rmed. Here, we present the main achievements of this collaboration in four main topics: 1) biology of the pest, 2) monitoring, 3) biological control, and 4) cooperative networks. Two mass rearing procedures have been implemented in the region with relative success, allowing basic biological studies on the pest. Continuous monitoring in the region for >5 years has provided a reasonable knowledge on seasonal patterns of T. peregrinus. Biological control strategies developed include the use of local natural enemies of T. peregrinus, the development of biopesticides, and the introduction of Cleruchoides noackae, an egg parasitoid of T. peregrinus from Australia. We review the main achievements in each country. Finally, a regional network of institutions, researchers, and students has strengthened in the region, providing a solid background for future collaboration.

Biological control of the bronze bug, Thaumastocoris peregrinus, in eucalyptus plantations in Brazil

The bronze bug Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae) was detected in Brazil in 2008 and infested >180 000 ha of eucalyptus plantations in 2011. The bronze bug can cause a reduction of 10–15% in wood productivity after 2 years of heavy infestation. Although there is not an effective control method known, biological control is the main control strategy studied. An exotic egg parasitoid, Cleruchoides noackae (Hymenoptera: Mymaridae), was imported from Australia in 2012, reared in a laboratory and released in three Brazilian regions. Parasitoids were recovered at release points after 20–30 d. In 2013, preliminary evaluations demonstrated parasitoid establishment in these areas, and the parasitoid was recovered in adjacent areas after 1 year of release. Bioassays confi rmed egg parasitism of 15–20% by C. noackae. Other native natural enemies were studied. We found green lacewing Chrysoperla externa and predatory bugs Supputius cincticeps and Atopozelus opsimus preying on nymphs and/or adults of T. peregrinus. Another promising possibility is entomopathogenic fungi. Commercial formulations of Beauveria bassiana were tested with success in lab and fi eld conditions. Fusarium proliferatum and Paecilomyces cateniannulatus caused mortality of T. peregrinus in natural epizooties. After 5 years of research, it is possible to develop an integrated pest management system (IPM) for eucalyptus plantations based on biocontrol strategy for bronze bug.

Harbor Porpoise (Phocoena phocoena) Abundance in Alaska: Bristol Bay to Southeast Alaska, 1991-1993

Between 1991 and 1993, Alaska harbor porpoise (Phocoena phocoena) abundance was investigated during aerial surveys throughout much of the coastal and offshore waters from Bristol Bay in the eastern Bering Sea to Dixon Entrance in Southeast Alaska. Line-transect methodology was used, and only those observations made during optimal conditions were analyzed. Survey data indicated densities of 4.48 groups/100 km2, or approximately 3,531 harbor porpoises (95% C.I. 2,206-5,651) in Bristol Bay and 0.54 groups/100 km2, or 136 harbor porpoises (95% C.I. 11-1,645) for Cook Inlet. Efforts off Kodiak Island resulted in densities of 1.85 groups/100 km2, or an abundance estimate of 740 (95% C.I. 259-2,115). Surveys off the south side of the Alaska Peninsula found densities of 2.03 groups/100 km2 and an abundance estimate of 551 (95% C.I. 423-719). Surveys of offshore waters from Prince William Sound to Dixon Entrance yielded densities of 4.02 groups/100 km’ and an abundance estimate of 3,982 (95% C.I. 2,567-6,177). Combining all years and areas yielded an uncorrected density estimate of 3.82 porpoises per 100 km2, resulting in an abundance estimate of 8,940 porpoises (CV = 13.8%) with a 95% confidence interval of 6,746-11,848. Using correction factors from other studies to adjust for animals missed by observers, the total number of Alaska harbor porpoises is probably three times this number.

Radiocarbon Dating, Chronologic Framework, and Changes in Accumulation Rates of Holocene Estuarine Sediments from Chesapeake Bay

Rapidly accumulating Holocene sediments in estuaries commonly are difficult to sample and date. In Chesapeake Bay, we obtained sediment cores as much as 20min length and used numerous radiocarbon ages measured by accelerator mass spectrometry methods to provide the first detailed chronologies of Holocene sediment accumulation in the bay. Carbon in these sediments is a complex mixture of materials from a variety of sources. Analyses of different components of the sediments show that total organic carbon ages are largely unreliable, because much of the carbon (including coal) has been transported to the bay from upstream sources and is older than sediments in which it was deposited. Mollusk shells (clams, oysters) and foraminifera appear to give reliable results, although reworking and burrowing are potential problems. Analyses of museum specimens collected alive before atmospheric nuclear testing suggest that the standard reservoir correction for marine samples is appropriate for middle to lower Chesapeake Bay. The biogenic carbonate radiocarbon ages are compatible with 210Pb and 137Cs data and pollen stratigraphy from the same sites. Post-settlement changes in sediment transport and accumulation is an important environmental issue in many estuaries, including the Chesapeake. Our data show that large variations in sediment mass accumulation rates occur among sites. At shallow water sites, local factors seem to control changes in accumulation rates with time. Our two relatively deep-water sites in the axial channel of the bay have different long-term average accumulation rates, but the history of sediment accumulation at these sites appears to reflect overall conditions in the bay. Mass accumulation rates at the two deep-water sites rapidly increased by about fourfold coincident with widespread land clearance for agriculture in the Chesapeake watershed.

Late Pliocene diatoms in a diatomite from Prydz Bay, East Antarctica

Very well-preserved Pliocene diatoms from a diatomite unit interbedded within glacial sediments at Ocean Drilling Program Site 742 in Prydz Bay, Antarctica are documented and illustrated. The presence of Thalassiosira kolbei, T torokina, Actinocyclus actinochilus, A karstenii and the absence of Nitzschia interfrigidaria, T. insigna and T. vulnifica in Sample 119-742A-15R-4, 44-46 cm constrain its age to ca. 2.2-1.8 Ma (late Pliocene). Diatoms associated with sea ice constitute 35% of the Pliocene diatom assemblage, compared with 71% of the modern sediment assemblage at the site, suggesting that sea ice was present during the late Pliocene period of deposition of the sample, although it probably was not the significant feature it is today.

BENTHIC MACROFAUNA STRUCTURE IN THE NORTHEAST AREA OF TODOS OS SANTOS BAY, BAHIA STATE, BRAZIL: PATTERNS OF SPATIAL AND SEASONAL DISTRIBUTION

Structure of intertidal and subtidal benthic macrofauna in the northeastern region of Todos os Santos Bay (TSB), northeast Brazil, was investigated during a period of two years. Relationships with environmental parameters were studied through uni- and multivariate statistical analyses, and the main distributional patterns shown to be especially related to sediment type and content of organic fractions (Carbon, Nitrogen, Phosphorus), on both temporal and spatial scales. Polychaete annelids accounted for more than 70% of the total fauna and showed low densities, species richness and diversity, except for the area situated on the reef banks. These banks constitute a peculiar environment in relation to the rest of the region by having coarse sediments poor in organic matter and rich in biodetritic carbonates besides an abundant and diverse fauna. The intertidal region and the shallower area nearer to the oil refinery RLAM, with sediments composed mainly of fine sand, seem to constitute an unstable system with few highly dominant species, such as Armandia polyophthalma and Laeonereis acuta. In the other regions of TSB, where muddy bottoms predominated, densities and diversity were low, especially in the stations near the refinery. Here the lowest values of the biological indicators occurred together with the highest organic compound content. In addition, the nearest sites (stations 4 and 7) were sometimes azoic. The adjacent Caboto, considered as a control area at first, presented low density but intermediate values of species diversity, which indicates a less disturbed environment in relation to the pelitic infralittoral in front of the refinery. The results of the ordination analyses evidenced five homogeneous groups of stations (intertidal; reef banks; pelitic infralittoral; mixed sediments; Caboto) with different specific patterns, a fact which seems to be mainly related to granulometry and chemical sediment characteristics.

CAPTURE FISHERY IN NORTHERN TODOS OS SANTOS BAY, TROPICAL SOUTHWESTERN ATLANTIC, BRAZIL

Information on marine and estuarine capture fishery activity in northern Todos os Santos Bay, northeastern Brazil, based on daily data collected between September 2003 and June 2005 is presented. Small-scale artisanal fishery in this area includes the use of traditional vessels both non-motorized and motorized for locomotion, being carried out mainly by canoe or on foot, and involves many different kinds of gear, including gillnet, hook and line, seine nets, and traps. A total of 113 taxa were grouped into 77 resources, including 88 fish, 10 crustaceans, and 15 mollusks. Data on nominal catches of fish, crustaceans and mollusks are presented by month and location. A total of 345.2 tonnes of fishery resources were produced (285.4 tonnes of fish, 39.2 tonnes of fresh invertebrates, and 20.6 tonnes of processed invertebrates). Temporal variation in the fish catch was associated with the life cycle of the species or with the hydrographic conditions. The first-sale value of this catch amounted to around US$ 615,000.00, fishes representing 71.3% of it. A table of the average price of each fishery resource is presented. The results produced in this study may be considered a reference for future monitoring programs of fishery resources in the area.

ICHTHYOFAUNA IN AN ESTUARY OF THE MATARIPE AREA, TODOS OS SANTOS BAY, BAHIA, BRAZIL

The community structure and dynamics as well as some biological parameters of selected species of the ichthyofauna of the Mataripe estuarine region affected by the Landulfo Alves Oil Refinery (RLAM) were analyzed. Twenty stations were sampled with a gillnet in five different periods: August and December 2003, March and July/August 2004 and January 2005. Thirty-five actinopterygian species and one elasmobranch species were recorded, Oligoplites saurus, Diapterus rhombeus, Lutjanus synagris and Scomberomorus brasiliensis among them, on all the campaigns. A total of 1368 specimens, weighing 36.10 kg, were caught. The ichthyofauna total biomass was greater, in weight, on the eastern side of the study region, especially at the stations close to the shoals/reefs and the rocky bottom. A similar pattern was also observed for the diversity values. In general, low evenness and diversity were observed in the area studied, possibly as a result of the fishing gear used. D. rhombeus juveniles dominated in all but one of the samplings (July 2004), in which latter Cyclichthys spinosus was dominant. Carangids and species associated with consolidated bottoms were observed, although in small numbers, throughout the study period. In spite of the limitations imposed by the gear used for sampling, the estuarine area influenced by the RLAM was seen to play a role as a growth area for the great majority of species, especially the mojarra (D. rhombeus), but it offers no fishing potential due to the prevalence of young and small individuals. Evidence of imminent spawning was recorded for Pomadasys corvinaeformis in August 2003, and recent spawning in March 2004 for Oligoplites saurus. Further, mature individuals occurred in insufficient numbers to permit population level evaluation.

Admiralty Bay Benthos Diversity-A census of a complex polar ecosystem

A thorough census of Admiralty Bay benthic biodiversity was completed through the synthesis of data, acquired from more than 30 years of observations. Most of the available records arise from successive Polish and Brazilian Antarctic expeditions organized since 1977 and 1982, respectively, but also include new data from joint collecting efforts during the International Polar Year (2007-2009). Geological and hydrological characteristics of Admiralty Bay and a comprehensive species checklist with detailed data on the distribution and nature of the benthic communities are provided. Approximately 1300 species of benthic organisms (excluding bacteria, fungi and parasites) were recorded from the bay`s entire depth range (0-500 m). Generalized classifications and the descriptions of soft-bottom and hard-bottom invertebrate communities are presented. A time-series analysis showed seasonal and interannual changes in the shallow benthic communities, likely to be related to ice formation and ice melt within the bay. As one of the best studied regions in the maritime Antarctic Admiralty Bay represents a legacy site, where continued, systematically integrated data sampling can evaluate the effects of climate change on marine life. Both high species richness and high assemblage diversity of the Admiralty Bay shelf benthic community have been documented against the background of habitat heterogeneity. (C) 2010 Elsevier Ltd. All rights reserved.

Bacterial diversity in rhizosphere soil from Antarctic vascular plants of Admiralty Bay, maritime Antarctica

The Antarctic is a pristine environment that contributes to the maintenance of the global climate equilibrium. The harsh conditions of this habitat are fundamental to selecting those organisms able to survive in such an extreme habitat and able to support the relatively simple ecosystems. The DNA of the microbial community associated with the rhizospheres of Deschampsia antarctica Desv (Poaceae) and Colobanthus quitensis (Kunth) BartI (Caryophyllaceae), the only two native vascular plants that are found in Antarctic ecosystems, was evaluated using a 16S rRNA multiplex 454 pyrosequencing approach. This analysis revealed similar patterns of bacterial diversity between the two plant species from different locations, arguing against the hypothesis that there would be differences between the rhizosphere communities of different plants. Furthermore, the phylum distribution presented a peculiar pattern, with a bacterial community structure different from those reported of many other soils. Firmicutes was the most abundant phylum in almost all the analyzed samples, and there were high levels of anaerobic representatives. Also, some phyla that are dominant in most temperate and tropical soils, such as Acidobacteria, were rarely found in the analyzed samples. Analyzing all the sample libraries together, the predominant genera found were Bifidobacterium (phylum Actinobacteria), Arcobacter (phylum Proteobacteria) and Faecalibacterium (phylum Firmicutes). To the best of our knowledge, this is the first major bacterial sequencing effort of this kind of soil, and it revealed more than expected diversity within these rhizospheres of both maritime Antarctica vascular plants in Admiralty Bay, King George Island, which is part of the South Shetlands archipelago. The ISME Journal (2010) 4, 989-1001; doi:10.1038/ismej.2010.35; published online 1 April 2010

SPATIAL AND TEMPORAL DISTRIBUTION OF FISH IN PALMAS BAY, UBATUBA, BRAZIL

The aim of this study was to analyze the distribution and abundance of the fish fauna of Palmas bay on Anchieta Island in southeastern Brazil. Specimens were caught in the summer and winter of 1992, using an otter trawl at three locations in the bay. The specimens were caught in both the nighttime and daytime. Data on the water temperature and salinity were recorded for the characterization of the predominant water mass in the region, and sediment samples were taken for granulometric analysis. A total of 7 656 specimens (79 species), with a total weight of approximately 300 kg, were recorded. The most abundant species were Eucinostomus argenteus, Ctenosciaena gracilicirrhus, Haemulon steindachneri, Eucinostomus gula and Diapterus rhombeus, which together accounted for more than 73% of the sample. In general, the ecological indices showed no differences in the composition of species for the abiotic variables analyzed. The multivariate analysis showed that the variations in the distribution of the fish fauna were mainly associated with intra-annual differences in temperature and salinity, resulting from the presence of South Atlantic Central Water (SACW) in the area during the summer. The analysis also showed an association with the type of bottom and a lesser association with respect to the night/day periods.