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.

Nutritional status of the cauliflower cultivar ‘verona’ grown with omission of out added macronutrients

Knowledge of plant nutritional status allows an understanding of the physiological responses of plants to crop fertilization. A hydroponic experiment evaluated the symptoms of macronutrient deficiency in cauliflower 'Verona'and determined: a) the macronutrient contents of foliar tissues when visual symptoms were observed, b) macronutrients content of foliar and inflorescence tissues at harvest. The effect of nutrient deficiency on inflorescence mass was also evaluated. Nitrogen deficiency caused chlorosis followed by purple color in the old leaves, while P deficiency caused only chlorosis in old leaves. Chlorosis at the edge of old leaves progressing to the center of the leaves was observed with the omission of K, and after was observed necrosis in the chlorotic areas. Ca deficiency caused tip burn in new leaves, while Mg deficiency caused internerval chlorosis in old leaves. The omission of eachmacronutrient reduced inflorescence dry matter. This deleterious effect was larger for N, P, and K deficiencies, reducing inflorescence dry matter by 87, 49, and 42%, respectively. When the nutrient solutions without N, P, K, Ca, or Mg were supplied to cauliflower plants, the macronutrient contents at harvest were 8.8, 0.6, 3.5, 13.0, and 0.8 g kg(-1) in the foliar tissues and 27.3, 2.2, 21.6, 1.1, and 0.7 g kg(-1) in the inflorescence tissues, respectively.

Gallus gallus domesticus are resistant to infection with Neospora caninum tachyzoites of the NC-1 strain

The aim of this study was to experimentally evaluate infection in Gallus gallus domesticus with Neospora caninum tachyzoites of the NC-1 strain. Experimental infection was conducted in 90-day-old chickens, embryonated eggs and bioassays in dogs. In the first experiment, poults were randomly divided into four groups. Groups I and II were provided feed with coccidiostat, whereas groups III and IV received feed without coccidiostat. When the poults from groups I and III reached 90 days of age, they received a subcutaneous inoculation of N. caninum. Once the hens entered their egg-laying period, during the following 30 days, the eggs were collected, identified, weighed and placed in an incubator. On the 70th day after inoculation, all animals, including the chicks, were euthanized. Tissue samples from the adult poultry and chicks were collected for histopathology, immunohistochemistry (IHC) and PCR. Brain tissue and pectoral muscle samples from infected birds were fed to two dogs. Notably, the average weight of the group III eggs was lower than that of the group IV eggs (p <0.05). No changes consistent with infection in adult poultry or chicks were detected by histopathology or IHC; moreover, no amplified parasite DNA was detected in the birds'tissues or dogs'feces. No dog eliminated oocysts. In the second experiment, the embryonated chicken eggs were inoculated with 1 x 10(2) N. caninum tachyzoites, on the 10th day of incubation, and chicks born from these eggs were housed in boxes suitable for the species and received commercial feed and distilled water ad libitum. On the 30th day after infection (DAI), the poultry were euthanized, and their organs were processed as described in experiment I. The amplification of parasite DNA was observed in the spleen and pectoral muscles of one of the birds. The ingestion of bird tissues by dogs did not result in oocyst elimination. These results indicate that the parasite may have been eliminated by the host and that the use of tachyzoites to induce chronic disease might be a poor source for hens. (C) 2014 Elsevier B.V. All rights reserved.

Identities among actin-encoding cDNAs of the Nile tilapia (Oreochromis niloticus) and other eukaryote species revealed by nucleotide and amino acid sequence analyses

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

Calcium hydroxide root canal sealers - Histopathologic evaluation of apical and periapical repair after endodontic treatment

Eighty root canals of the premolars of 4 dogs, with vital pulp, were instrumented and filled during the same session with the Sealapex, CRCS, Sealer 26, and Apexit sealers, the animals were sacrificed 180 days after root canal filling and their maxillae and mandibles were removed and fixed in 10% formalin, After routine histologic processing, the sections were stained with hematoxylin-eosin and Mallory trichrome, Histopathologic analysis showed that Sealapex was the sealer that best permitted the deposition of mineralized tissue at the apical level and was the only sealer that provided complete sealing (37.5% of cases), With the use of Sealapex, no inflammatory infiltrate occurred and there was no reabsorption of mineralized tissues, In contrast, partial sealing and a moderate inflammatory infiltrate occurred with the use of CRCS, When Apexit and Sealer 26 were used the absence of sealing was frequent and active reabsorption of mineralized tissues occurred in most cases, the inflammatory infiltrate predominating with the use of Apexit was of the severe type, whereas with the use of Sealer 26 the inflammatory infiltrate was mild or absent.

Spindle cell lipoma occurring in the buccal mucosa: an unusual location of this benign lipomatous neoplasm

Spindle cell lipoma is a benign lipomatous neoplasm, which rarely occurs in the oral cavity. The aims of this paper are to report a case of spindle cell lipoma located in buccal mucosa and discuss the main clinical, histological, and immunohistochemical findings of this entity. Thus, we report a 4-year history of an asymptomatic smooth surface nodule in an elderly Caucasian man with clinical hypothesis of fibroma. The histopathological examination showed spindle cells, mature adipose tissue, and many mast cells in a stroma of connective tissue presenting ropey collagen fibers bundles. After immunohistochemical analysis, the final diagnosis was spindle cell lipoma.

Long-term evaluation of oral gavage with periodontopathogens or ligature induction of experimental periodontal disease in mice

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

Estudo neuroanatômico do éncefalo bovino por ressonância magnética

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Histologic and histometric evaluation of calvarial autogenous bone graft in reconstruction of atrophyc maxilla for dental implants

The calvarial bone is highlighted as a good donor area for large reconstructions of atrophic jaw for subsequent rehabilitation with implant-supported prosthesis. The aim of this study was to observe and measure through histological and histometric evaluation, the cellular events that occur at the interface of union from onlay parietal bone graft on the maxilla of 10 patients, after a period of 6 months of incorporation. The biopsies were performed at the time of installation of osseointegrated implants. The bone contact area represented 78.75% and connective contact 21.25%. The region of connective union between the bone graft to the maxillae presented new bone formation (41.26%), marrow bone (36.06%), osteoid tissue (15.86%) and connective tissue (6.80%). All samples had good graft incorporation to the receptor bed with osteogenic activity and absence of inflammatory cells.

Susceptibility of microorganisms isolated from necrotic pulps to calcium hydroxide

Calcium hydroxide has been used as pulp-capping agent and canal dressing due to its antimicrobial and anti-inflammatory properties besides its ability to induce formation of mineralized tissues. The aim of this study was to evaluate the susceptibility to calcium hydroxide of 146 bacterial strains isolated from endodontic infections. MIC was determined by using an agar dilution method, while contact bactericide activity was performed through in broth. All the isolates were sensitive to calcium hydroxide in concentrations that varied from 0.5mg/ml to 128 mg/ml, and the genera Enterococcus, Pseudomonas, Staphylococcus and Actinomyces were the most resistant. Gramnegative anaerobes proved to be the most sensitive isolates. All the isolates were inhibited after 60 minutes of contact with the alkali in concentration of 100mg/ml

The Pathologic Copepod Phrixocephalus cincinnatus (Copepoda: Pennellidae) in the Eye of Arrowtooth Flounder, Atherestes stomias, and Rex Sole, Glyptocephalus zachirus, from British Columbia

We report Phrixocephalus cincinnatus, a pennellid copepod infecting the eyes of flatfishes, from a single specimen of rex sole, Glyptocephalus zachirus, for the first time. In the typical host, the arrowtooth flounder, Atherestes stomias, the parasite occurred commonly in sampled populations from the Broughton Archipelago in British Columbia, infected primarily the right eye of the flounder, and on only one occasion presented more than two parasites per eye. The copepod attached to the choroid layer and ramified throughout the posterior compartment of the eye, resulting in the disruption of the retina and probably impairing host vision. Inflammation and hyperplasia progressed to necrosis and proliferation of connective tissue, resulting in the total destruction of the eye.

Virus-Associated Tumours of the Harderian Gland in Laboratory-Reared Voles, Clethrionomys rutilis (Pallas)

Tumors of the Harderian gland occurred frequently in red-backed voles, Clethrionomys rutilus (Pallas), in a captive colony maintained since 1957. Although the tumors became obvious only in animals about a year old and older, early stage changes were histologically evident in glands from younger, clinically normal voles. The tumors ranged in type from poorly differentiated adenoma to adenocarcinoma, with invasion of adjacent tissues and characteristic deformation of the zygomata. Electron micrographs disclosed viral particles in the cytoplasm of glands from clinically normal animals as well as in glands showing malignant changes. The tumor was not observed in arvicoline rodents of other species maintained in the same rooms with the red-backed voles.

GaAs 904-nm laser irradiation improves myofiber mass recovery during regeneration of skeletal muscle previously damaged by crotoxin

This work investigated the effect of gallium arsenide (GaAs) irradiation (power: 5 mW; intensity: 77.14 mW/cm(2), spot: 0.07 cm(2)) on regenerating skeletal muscles damaged by crotoxin (CTX). Male C57Bl6 mice were divided into six groups (n = 5 each): control, treated only with laser at doses of 1.5 J or 3 J, CTX-injured and, CTX-injured and treated with laser at doses of 1.5 J or 3 J. The injured groups received a CTX injection into the tibialis anterior (TA) muscle. After 3 days, TA muscles were submitted to GaAs irradiation at doses of 1.5 or 3 J (once a day, during 5 days) and were killed on the eighth day. Muscle histological sections were stained with hematoxylin and eosin (H&E) in order to determine the myofiber cross-sectional area (CSA), the previously injured muscle area (PIMA) and the area density of connective tissue. The gene expression of MyoD and myogenin was detected by real-time PCR. GaAs laser at a dose of 3 J, but not 1.5 J, significantly increased the CSA of regenerating myofibers and reduced the PIMA and the area density of intramuscular connective tissue of CTX-injured muscles. MyoD gene expression increased in the injured group treated with GaAs laser at a dose of 1.5 J. The CTX-injured, 3-J GaAs laser-treated, and the CTX-injured and treated with 3-J laser groups showed an increase in myogenin gene expression when compared to the control group. Our results suggest that GaAs laser treatment at a dose of 3 J improves skeletal muscle regeneration by accelerating the recovery of myofiber mass.

Accumulation of six metals in the mangrove crab Ucides cordatus (Crustacea: Ucididae) and its food source, the red mangrove Rhizophora mangle (Angiosperma: Rhizophoraceae)

The crab Ucides cordatus and the red mangrove Rhizophora mangle are endemic mangrove species and potential bio-accumulators of metals. This study quantified the accumulation of six metals (Cd, Cr, Cu, Hg, Mn and Pb) in different organs (claw muscle, hepatopancreas and gills) of U. cordatus, as well as in different maturation stages of the leaves (buds, green mature, and pre-abscission senescent) of R. mangle. Samples were collected from mangrove areas in Cubatao, state of Sao Paulo, a heavily polluted region in Brazil. Data for metal contents in leaves were evaluated by one-way ANOVA; while for crabs a factorial ANOVA was used to investigate the effect of different tissues, animal size and the interactions between them. Means were compared by Tukey test at five percent, and the association between the metal concentrations in each crab organ, depending on the size, was evaluated by Pearson's linear correlation coefficient (r). Concentrations of Pb and Hg were undetectable for the different leaf stages and crab tissues, while Cd concentrations were undetectable in the leaf stages. In general, the highest accumulation of metals in R. mangle leaves occurred in pre-abscission senescent and green mature leaves, except for Cu, which was found in the highest concentrations in buds and green mature leaves. For the crab, Cd, Cu, Cr and Mn were present in concentrations above the detection limit, with the highest accumulation in the hepatopancreas, followed by the gills. Cu was accumulated mostly in the gills. Patterns of bioaccumulation between the crab and the mangrove tree differed for each metal, probably due to the specific requirements of each organism for essential metals. However, there was a close and direct relationship between metal accumulation in the mangrove trees and in the crabs feeding on them. Tissues of R. mangle leaves and U. cordatus proved effective for monitoring metals, acting as important bioindicators of mangrove areas contaminated by various metals. (C) 2012 Elsevier Inc. All rights reserved.

Kidney of Minke Whale (Baleanoptera acutorostrata): Architecture and structure

Sarmento C. A. P., Ferreira A. O., Rodrigues E. A. F., Lesnau G. G., Rici R. E. G., Abreu D. K., Biasi C. & Miglino M. A. 2012. [Kidney of Minke Whale (Baleanoptera acutorostrata): Architecture and structure.] Rins de Baleia Minke (Baleanoptera acutorostrata): arquitetura e estrutura. Pesquisa Veterinaria Brasileira 32(8): 807-811. Departamento de Cirurgia, Setor de Anatomia dos Animais Domesticos e Silvestres, Universidade de Sao Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Sao Paulo, SP 05508-270, Brazil. E-mail: sarmento@usp.br Among marine mammals, whale is one of the most attention-arousing animals, especially concerning its urinary tract. This system follows the pattern of mammals with regard to its constitution, however, it differs in renal morphology and number of lobes, which, in turn, form complete reniculi, agglutinated in hundreds. This structure is supported by fibrous connective tissue, but highly capable of maintaining electrolyte balance. Six pairs of kidneys of Minke whale (Balaenoptera acutorostrata), collected in 1982, in Cabedelo, Paraiba, Brazil, in the last fishing allowed, were dissected. These kidneys were preserved in 10% formaldehyde and they presented a very large histologic layer of collagen surrounding the medullary wall. The urinary collecting duct form papillary glasses, that reach a single collecting center which discharges in the ureter. It was found that the kidney of Minke whale has a lobe characteristic, with, on average, 700 reniculi; each reniculus has anatomical and functional characteristics of a unipyramidal kidney, with an inner layer (medulla), and an outer layer (cortex), and independent irrigation, with formation of individually arcuate arteries, as observed in unipyramidal terrestrial mammals. However, the set gathering all these reniculi constitutes, in the end, a multilobular and polipyramidal kidney, contrary to the morphology of most terrestrial mammals. It was not possible to distinguish the renicular cortex structures of the Minke whale in the level of light microscopy. Through scanning electron microscopy, it was possible to visualize a cortical layer located between two fibrous capsules. This joint, in turn, consists of connective tissue, which, along with a layer of collagen and elastic fibers, separates the cortex from the medulla; the kidney glomeruli were visualized, completely taken by the glomerular vessels and arranged into several layers. One notices that the glomerular cavity is almost a virtual space into which the glomerular filtrate is drained, and it does not present a globular shape. Vascularization is increased in the medullary region. The difference between the kidneys of terrestrial and marine mammals consists in the arrangement of morphological components, favoring the organ's physiology.