Transcriptional and cellular effects of paracetamol in the oyster Crassostrea gigas.

Acetaminophen (paracetamol) (PAR) is one of the most popular non-steroidal anti-inflammatory drugs (NSAIDs) with analgesic and antipyretic properties consumed worldwide and often detected in the aquatic environment. Due to the fact that PAR induces oxidative stress in mammals, the aim of this study was to evaluate if similar effects were observed in oysters Crassostrea gigas, given their economic and ecological importance and worldwide distribution. Oysters were exposed for 1, 4 and 7 days to two different sublethal PAR concentrations (0, 1 and 100µgL-1). Cell viability, DNA damage in hemocytes and enzymatic activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidases (GPx), glutathione reductase (GR), glucose 6-phosphate dehydrogenase (G6PDH) and glutathione S-transferases (GST) were evaluated in oyster gills. In addition, changes at transcriptional level of Cu/Zn superoxide dismutase (SOD), catalase-like (CAT-like), cytochrome P450 genes (CYP30C1, CYP2AU2, CYP3071A1, CYP356A1), glutathione S-transferase isoforms (GST-ω and GST-π-like), cyclooxygenase (COX), fatty acid binding proteins-like (FABP-like), and caspase genes were evaluated in oyster gills and digestive gland. No changes in cell viability and DNA damage were observed in oysters exposed to both PAR concentrations. Similarly, no significant changes were detected in the major antioxidant enzymes (except for auxiliary enzyme GR) in oyster gills, suggesting that changes in GR activity are enough to counteract a potential oxidative stress in C. gigas gills under these experimental conditions. Furthermore, changes at transcriptional level are concentration and tissue dependent. PAR elicited an inhibition of CYP30C1, CYP3071A1 and FABP-like transcripts highlighting their role in drug metabolism, transport and detoxification of PAR in the gills. GST transcript levels were type, tissue and concentration-dependent. GST-π-like was down-regulated in oyster gills exposed to the lowest PAR concentration and up-regulated in the digestive gland of oysters exposed to the highest PAR concentration. However, GST-ω transcript levels were lower only in oysters digestive gland exposed to the lowest PAR concentration. Therefore, changes at transcriptional level were more sensitive to assess the exposure to PAR at environmental relevant concentrations.

The reproductive cycle of the oyster Crassostrea gasar.

The purpose of this study was to analyze the reproductive cycle of the oyster Crassostrea gasar (= C. brasiliana) in the field and the laboratory. The reproductive cycle of the animals was evaluated in the field at Sambaqui Beach, Florianópolis, SC (27° 29'18″ S and 48° 32'12″ W) from May 2008 through November 2009. In July, the animals were in the resting stage. The early growth stage began in August and was followed by the late growth stage in October. In November and December, the oysters began to enter the mature stage. Females in spawning condition were predominant during these months. The stages of the reproductive cycle were positively associated with temperature (r=0.77, P<0.01) and negatively associated with salinity (r=-0.56, P=0.042). These findings demonstrated that increased temperature and reduced salinity influence the reproductive development of Crassostrea gasar. The condition index (CI) of the animals was also associated with the seawater temperature. The highest values of the condition index were observed during the months when the temperature of the seawater was gradually increasing. A laboratory experiment was performed to test the effect of salinity on the reproductive cycle of the oysters. The experiment was conducted in standardized tanks. The animals were conditioned using two salinities (24‰ and 34‰). The salinity regime influenced the development of the gonadal tissue of the oysters. A salinity of 24‰ produced greater reproductive development.

Growth and survival of the pearl oyster Pteria hirundo (L.,1758) in an intermediate stage of culture in Santa Catarina, Brazil.

There are no records in Brazil for the culture of Pteridae family oysters, genus Pteria and Pinctada. The intermediate culture can be considered one of the critical stages during the oyster farming. The changes in life conditions from a semi-closed environment provided by the collector to the structures utilized for the growth represents a considerable stress factor to which the animals are subjected. The goal of the present study was to evaluate the growth and survival of the pearl oyster Pteria hirundo (Linnaeus, 1758) kept in intermediate lanterns. Seeds of P. hirundo, obtained through larviculture carried out at the Marine Molluscs Laboratory (UFSC), southern Brazil were detached from the collectors, transferred and kept in an intermediate culture system (oyster lantern). During 6 months, growth (dorso-ventral height - DVH in mm) and survival (%) were evaluated. Two size classes were tested: medium (M), between 3.4 and 4.4 mm, and large (L), bigger than 4.5 mm. The density of occupation in each floor of the lantern was 50%. At the end of the experiment, the survivals observed were 90 and 94% for the sizes M and L, respectively. Medium juveniles of Pteria hirundo had an average (Std) increase of 16.1 (±4.38) mm in the dorso-ventral height, and large juveniles an average increase of 11.4 (±3.77) mm after six months of experiment. The results observed during the intermediate stage of cultivation indicate that Pteria hirundo presents survival and growth similar to other species of pearl oysters traditionally used in the pearl industry in several countries.

Larval settlement and spat recovery rates of the oyster Crassostrea brasiliana (Lamarck, 1819) using different systems to induce metamorphosis.

This study aimed at the assessment, in the laboratory, of the larval settlement and spat recovery rates of oysters of the species Crassostrea brasiliana using plastic collectors, epinephrine (C9H13NO3 C4H6O6) and shell powder in settlement tanks. Polypropylene was used attached to bamboo frames. The material was chosen due to its pliability--that favours the spat detachment. Two experiments were carried out; the first between February and April 2008, and the second between November and December 2008 at the Marine Mussel Laboratory of Santa Catarina Federal University (Laboratório de Moluscos Marinhos da Universidade Federal de Santa Catarina). In the first experiment, the scratched plastic collectors were tested consorting them with shell powder; on the second, the plastic collectors were tested consorted with shell powder, only shell powder and epinephrine as the metamorphosis stimulator. The quantification was carried out of the larvae settled in the plastic collectors, and of the recovery and integrity of the spats after their detachment. The first experiment has shown a recovery rate of 48.83% of the spats in comparison with the D larvae used. From this percentage, 4.9% settled in the plastic collectors and 43.93% in shell powder. The second experiment revealed 55.78% regarding the settled spats in comparison with the total of larvae used (using epinephrine), 78.62% in the treatment with the collector plus shell powder and 58.33% in the treatment only with shell powder. Thus, the use of the collector plus shell powder resulted in a greater spat recovery when compared to the other treatments.

Infecção por cestóides proteocefalídeos em tucunaré Cichla sp. (Osteichthyes: Cichlidae), no Rio Paraná, São Paulo / Proteocephalid cestode infection in tucunaré Cichla sp. (Osteichthyes: Cichlidae) from Paraná River, São Paulo

The occurrence of proteocephalid cestodes in tucunaré Cichla sp., captured monthly, between August 2000 and August 2001, in Paraná River, Presidente Epitácio, SP, was evaluated. From 128 specimens, 71 (55.6 percent) were parasitized by Proteocephalus macrophallus (Diesing, 1850) and/or P. microscopicus (Woodland, 1935). Total mean abundance and intensity were 157.08 and 223.41, respectively. The highest prevalence (90 percent) mean abundance (1,122.4) and intensity indexes (1,247.11) occurred in February 2001, while in September 2000 there were no observed animals infected by cestodes. No relationship between the sex of the host and parasitological indexes was found.

Avaliou-se a ocorrência de cestóides proteocefalídeos em tucunaré Cichla sp., capturados mensalmente, entre agosto de 2000 e agosto de 2001, no rio Paraná, em Presidente Epitácio, SP. Um total de 128 espécimes foram analisados, dos quais 71 (55,6 por cento) estavam parasitados por Proteocephalus macrophallus (Diesing, 1850) e/ou P. microscopicus (Woodland, 1935). A abundância e intensidade média total foram de 157,08 e 223,41, respectivamente. A maior prevalência (90 por cento), juntamente com os maiores índices de abundância (1122,4) e intensidade média (1247,11) ocorreram no mês de fevereiro 2001, enquanto no mês de setembro 2000 não foram observados animais parasitados por cestóides. Não houve relação entre o sexo do hospedeiro e os índices parasitológicos.

Gene expression profile of the plant pathogen Fusarium graminearum under the antagonistic effect of Pantoea agglomerans.

The pathogenic fungus Fusarium graminearum is an ongoing threat to agriculture, causing losses in grain yield and quality in diverse crops. Substantial progress has been made in the identification of genes involved in the suppression of phytopathogens by antagonistic microorganisms; however, limited information regarding responses of plant pathogens to these biocontrol agents is available. Gene expression analysis was used to identify differentially expressed transcripts of the fungal plant pathogen F. graminearum under antagonistic effect of the bacterium Pantoea agglomerans. A macroarray was constructed, using 1014 transcripts from an F. graminearum cDNA library. Probes consisted of the cDNA of F. graminearum grown in the presence and in the absence of P. agglomerans. Twenty-nine genes were either up (19) or down (10) regulated during interaction with the antagonist bacterium. Genes encoding proteins associated with fungal defense and/or virulence or with nutritional and oxidative stress responses were induced. The repressed genes coded for a zinc finger protein associated with cell division, proteins containing cellular signaling domains, respiratory chain proteins, and chaperone-type proteins. These data give molecular and biochemical evidence of response of F. graminearum to an antagonist and could help develop effective biocontrol procedures for pathogenic plant fungi.

Chicken skeletal muscle-associated macroarray for gene discovery.

Macro- and microarrays are well-established technologies to determine gene functions through repeated measurements of transcript abundance. We constructed a chicken skeletal muscle-associated array based on a muscle-specific EST database, which was used to generate a tissue expression dataset of ~4500 chicken genes across 5 adult tissues (skeletal muscle, heart, liver, brain, and skin). Only a small number of ESTs were sufficiently well characterized by BLAST searches to determine their probable cellular functions. Evidence of a particular tissue-characteristic expression can be considered an indication that the transcript is likely to be functionally significant. The skeletal muscle macroarray platform was first used to search for evidence of tissue-specific expression, focusing on the biological function of genes/transcripts, since gene expression profiles generated across tissues were found to be reliable and consistent. Hierarchical clustering analysis revealed consistent clustering among genes assigned to 'developmental growth', such as the ontology genes and germ layers. Accuracy of the expression data was supported by comparing information from known transcripts and tissue from which the transcript was derived with macroarray data. Hybridization assays resulted in consistent tissue expression profile, which will be useful to dissect tissue-regulatory networks and to predict functions of novel genes identified after extensive sequencing of the genomes of model organisms. Screening our skeletal-muscle platform using 5 chicken adult tissues allowed us identifying 43 'tissue-specific' transcripts, and 112 co-expressed uncharacterized transcripts with 62 putative motifs. This platform also represents an important tool for functional investigation of novel genes; to determine expression pattern according to developmental stages; to evaluate differences in muscular growth potential between chicken lines, and to identify tissue-specific genes.

The significance of cortisol on acclimation to salinity in pejerrey Odontesthes bonariensis

The role of cortisol on the osmoregulation of pejerrey Odontesthes bonariensis at different salinities was investigated in adult fish injected with 0.7mg hydrocortisone per 100g body weight of fish, and transferred to 0, 5 and 20ppt of NaCl. Blood cortisol was 566ng/ml at the beginning of the experiment (0h) but surged to 1250ng/ml within 3h in cortisol-injected fish. Cortisol levels were influenced not only by treatment but also by time, being higher at 3h compared to 24h. Salinity level, time of exposure and their interaction, but not cortisol treatment, significantly affected plasma osmolality and the concentration of ions Cl- and Na+. This study showed that exogenous cortisol does not seem to play a significant role on the regulation of plasma osmolality and concentration of individual ions in pejerrey

Investigou-se a participação do cortisol na osmoregulação de peixe-rei Odontesthes bonariensis, em diferentes salinidades, em peixes adultos injetados com 0,7mg hidrocortisona por 100g de peso corporal, e transferidos para 0, 5 e 20ppt de NaCl. No inicio do experimento (0h), o cortisol encontrado no plasma foi de 566ng/ml, aumentando para 1250ng/ml em 3h em peixes injetados com cortisol. A concentração de cortisol foi influenciada não somente pelo tratamento, mas também pelo tempo, sendo maior 3h após a inoculação, comparada à 24h. A salinidade, o tempo de exposição e a interação desses dois fatores, mas não o tratamento com o cortisol, afetaram significativamente a osmolaridade e a concentração dos íons Cl- e Na+ do plasma. Este estudo mostrou que, o cortisol exógeno não influi significativamente na regulação da osmolaridade e da concentração de íons Na+ e Cl- no plasma em peixe-rei

Genetic parameters for test day milk yields of first lactation Holstein cows by random regression models.

Covariance components for test day milk yield using 263 390 first lactation records of 32 448 Holstein cows were estimated using random regression animal models by restricted maximum likelihood. Three functions were used to adjust the lactation curve: the five-parameter logarithmic Ali and Schaeffer function (AS), the three-parameter exponential Wilmink function in its standard form (W) and in a modified form (W*), by reducing the range of covariate, and the combination of Legendre polynomial and W (LEG+W). Heterogeneous residual variance (RV) for different classes (4 and 29) of days in milk was considered in adjusting the functions. Estimates of RV were quite similar, rating from 4.15 to 5.29 kg2. Heritability estimates for AS (0.29 to 0.42), LEG+W (0.28 to 0.42) and W* (0.33 to 0.40) were similar, but heritability estimates used W (0.25 to 0.65) were highest than those estimated by the other functions, particularly at the end of lactation. Genetic correlations between milk yield on consecutive test days were close to unity, but decreased as the interval between test days increased. The AS function with homogeneous RV model had the best fit among those evaluated.

Mapping QTLs on chicken chromosome 1 for performance and carcass traits in a broiler x layer cross.

With the objective of mapping quantitative trait loci (QTLs) for performance and carcass traits, an F2 chicken population was developed by crossing broiler and layer lines. A total of 2063 F2 chicks in 21 full-sib families were reared as broilers and slaughtered at 42 days of age. Seventeen performance and carcass traits were measured. Parental F(0) and F1 individuals were genotyped with 80 microsatellites from chicken chromosome 1 to select informative markers. Thirty-three informative markers were used for selective genotyping of F2 individuals with extreme phenotypes for body weight at 42 days of age (BW42). Based on the regions identified by selective genotyping, seven full-sib families (649 F2 chicks) were genotyped with 26 markers. Quantitative trait loci affecting body weight, feed intake, carcass weight, drums and thighs weight and abdominal fat weight were mapped to regions already identified in other populations. Quantitative trait loci for weights of gizzard, liver, lungs, heart and feet, as well as length of intestine, not previously described in the literature were mapped on chromosome 1. This F2 population can be used to identify novel QTLs and constitutes a new resource for studies of genes related to growth and carcass traits in poultry.