Salmonid fish viruses and cell interactions at early steps of the infective cycle

A flow cytometric virus-binding assay that directly visualizes the binding and entry of infectious pancreatic necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and virus haemorrhagic septicaemia virus (VHSV) to several cell lines was established. The highest efficiency of binding was shown by the BF-2 cell line and this was used to study, at the attachment level, the interactions of these cells with salmonid fish viruses in coinfections, and to further determine if the earliest stage of the viral growth cycle could explain the previously described loss of infectivity of IHNV when IPNV is present. Our results demonstrated that IPNV binds to around 88% of cells either in single or dual infections, whereas IHNV attachment always decreased in the presence of any of the other viruses. VHSV binding was not affected by IPNV, but coinfection with IHNV reduced the percentage of virus-binding cells, which suggests competition for viral receptors or co-receptors. Internalization of the adsorbed IHNV was not decreased by coinfection with IPNV, so the hypothetical competence could be restricted to the binding step. Treatment of the cells with antiviral agents, such as amantadine or chloroquine, did not affect the binding of IPNV and VHSV, but reduced IHNV binding by more than 30%. Tributylamine affected viral binding of the three viruses to different degrees and inhibited IPNV or IHNV entry in a large percentage of cells treated for 30 min. Tributylamine also inhibited IHNV cytopathic effects in a dose-dependent manner, decreasing the virus yield by 4 log of the 50% endpoint titre, at 10 mm concentration. IPNV was also inhibited, but at a lower level. The results of this study support the hypothesis that IHNV, in contrast to VHSV or IPNV, is less efficient at completing its growth cycle in cells with a simultaneous infection with IPNV. It can be affected at several stages of viral infection and is more sensitive to the action of antiviral compounds.     

Enriching rotifers with “premium” microalgae. Isochrysis aff. galbana clone T-ISO

The effect of semi-continuous culture on the nutritional value of microalgae was tested in the rotifer Brachionus plicatilis in short-term enrichment experiments. Isochrysis aff. galbana clone T-ISO was cultured semi-continuously with renewal rates from 10 to 50% of the volume of the culture per day and used to feed the rotifers. After 24 h, dramatic differences in dry weight and protein, lipid and carbohydrate contents were observed in the rotifers depending on the renewal rate applied to the microalgal culture. Rotifers fed T-ISO cultured with low renewal rates showed low dry weight and organic content, whereas rotifers fed microalgae from nutrient-sufficient, high renewal rate cultures showed higher dry weight and increases up to 60% in protein, 35% in lipid and 100% in carbohydrate contents. Feed conversion rate (FCR) and organic FCR decreased with increasing renewal rates, indicating a more efficient assimilation of the microalgal biomass obtained at high growth rates. The fatty acid profile of rotifers reflected that of T-ISO, with maximum content of polyunsaturated fatty acids (PUFAs), n-3 fatty acids and docosahexaenoic acid (DHA) being found in the rotifers fed microalgae from the renewal rate of 40%. Results demonstrate that the biochemical composition of B. plicatilis is strongly modified through the use of semi-continuous cultures of microalgae in short-term enrichment processes. This technique provides an excellent tool to improve the nutritional value of the live feed used in fish larvae cultures.     

Irrigation of paddy soil with industrial landfill leachate: impacts in rice productivity,plant nutrition,and chemical characteristics of soil

Treated industrial effluents have high levels of nutrients and dissolved organic matter. The irrigation of rice by flooding can increase nutrient uptake and grain yield. Therefore, this study evaluated the nutrient contents in the shoots and grain of the rice crop and also the chemical of the soil after irrigation of the crop with leachate of the treated industrial effluent. A greenhouse experiment was conducted using pots filled with 20 kg of soil in a randomized block design with three replications. The treatments consisted of control (irrigation with distilled water) and four concentrations of the leachate (25, 50, 75, and 100 %) for irrigation. At the end of the experiment, the nutrient contents in tissues of rice plants, sterility of spikelets, and grain mass were evaluated. Results showed that irrigation with the leachate at 25 % content increased the macro- and micronutrients’ concentrations in the shoot biomass and grain, except for potassium and iron. Irrigation with the industrial leachate decreased tillering and grain yield; however, it increased chlorophyll content, sterility of spikelets, and sodium intake at this leachate concentration. The potassium and sodium levels and the electrical conductivity values of soils irrigated with treated industrial leachate were increased. The use of the treated leachate from industrial effluents is an alternative that reuses the nutritional load, but the volume of leachate should be limited and monitored to prevent the sodicity in the soil and problemsdue to eutrophication.     

S‐locus diversity and cross‐compatibility of wild Prunus avium for timber breeding

Prunus avium is primarily cultivated for its fruit, sweet cherries. However, it is also used to produce high‐quality timber. In a P. avium seed orchard, gametophytic self‐incompatibility is a restriction for free pollen flow and should be considered when establishing basic forest materials. In this study, S‐locus diversity and cross‐incompatibility of wild cherry individuals in clonal banks established for breeding for timber production were investigated. Wild cherry trees (140) with outstanding forest growth habit, collected in northern Spain, grafted and planted in two clonal banks, were genotyped at the S‐locus. The self‐incompatibility S‐locus genes, S‐RNase and SFB, were analysed by PCR. Twenty‐two S‐haplotypes, resulting in 72 different S‐genotypes, were identified. The genotypes were grouped into 33 incompatibility groups and 39 unique genotypes. This initial S‐locus analysis revealed large genetic diversity of wild cherry trees from the Spanish northern deciduous forest, and provides useful information for seed orchard design. Wild P. avium displays significantly more genetic diversity than what is detected in local cultivars, revealing a narrowing of genetic diversity during local domestication.     

Two reported cytotypes of the emergent orchid model species <Emphasis Type="Italic">Erycina pusilla</Emphasis> are two different species

Each species is characterized by a specific set of chromosomes, which is described as the chromosome portrait or karyotype. In general, such a karyotype is the same for all individuals in the population. An exception to that rule has recently been found in the orchid Erycina pusilla, which has been reported to have two cytotypes with chromosome numbers of 2n = 10 and 2n = 12. Here, we examined the karyotypes of the two cytotypes and found differences in arm ratios and heterochromatin patterns as well as in the presence of satellite chromosomes and in the number and location of rDNA and telomeric repeat sites. These differences are extensive and would have required multiple chromosome rearrangements to generate the differences between the two karyotypes. We also found that F1 hybrids between the parents with the two different chromosome numbers resulted in sterile offspring, in accordance with our previous findings. The combination of hybrid sterility and extensively rearranged chromosomes supports the hypothesis that these two reported cytotypes are, in fact, two different species.     

Use of genomic and phenotypic selection in lines for prediction of testcrosses in maize II: grain yield and plant traits

Plant breeders have been trying to predict the performance of hybrids based on their parental performance. One application of molecular markers is its use in selection. The objectives were to map quantitative trait loci (QTL) and verify its congruence in maize lines and in their testcrosses and verify the possibility to select testcrosses from the predicted means of the lines by using information from markers. Two-hundred and fifty six lines and the testcrosses of these lines with two testers were evaluated in six environments, considering grain yield, plant lodging, days to anthesis and silking, anthesis-silking interval, plant and ear height and ear placement. QTL were mapped in the lines and in testcrosses and the predicted means of the lines were computed based on QTL effects and in all markers of the genome. The congruence of QTL detected in the lines and testcrosses were small for all traits. The correlations between the predicted means of the lines and the phenotypic means of the testcrosses ranged from low for grain yield to moderate for cycle and stature traits. The highest coincidences of the lines and selected testcrosses were observed for cycle and stature traits and the lowest for grain yield. Even by using molecular markers information, it is only possible to predict the testcrosses performance from the lines information to less complex traits and with reduced dominance effect. For complex traits and with pronounced dominance effect, information of markers must be obtained directly in the testcrosses, so they can be used for selection.     

Vacuum Drying for Extending Litchi Shelf-Life: Vitamin C,Total Phenolics,Texture and Shelf-Life Assessment

In an attempt to obtain shelf-stable litchi fruit with preserved nutritional quality and good sensory features, quarters of peeled and pitted fruits were vacuum dried at 50, 60 and 70 °C at a constant pressure of 8.0 kPa. The product was assessed for its vitamin C, total phenolics and texture (hardness). In addition, the product with the best texture was assessed for its shelf-life by means of accelerated testing. Results suggest that vacuum dried litchi retained almost 70% of the vitamin C and total phenolics when compared to frozen fruits (control). Vitamin C and phenolic compounds content significantly decreased with drying, while no difference was found between different drying temperatures. Hardness increased with drying temperature. The sample dried at 70 °C presented crispness, which is a desired quality feature in dried fruit products. This sample was subjected to shelf-life evaluation, whose result suggests a shelf-life of eight months at 23 °C. Total color change (CIE ΔE₀₀) was the expiry criterion. Vacuum drying was a suitable technique for producing shelf-stable litchi fruit with good texture while preserving its desirable original nutrients. Consumption of vacuum dried litchi may be beneficial to health due to its remarkable content of phenolic compounds and vitamin C.     

<Emphasis Type="Italic">Lupinus albus</Emphasis> Conglutin Gamma Modifies the Gene Expressions of Enzymes Involved in Glucose Hepatic Production <Emphasis Type="Italic">In Vivo</Emphasis>

Lupinus albus seeds contain conglutin gamma (Cγ) protein, which exerts a hypoglycemic effect and positively modifies proteins involved in glucose homeostasis. Cγ could potentially be used to manage patients with impaired glucose metabolism, but there remains a need to evaluate its effects on hepatic glucose production. The present study aimed to analyze G6pc, Fbp1, and Pck1 gene expressions in two experimental animal models of impaired glucose metabolism. We also evaluated hepatic and renal tissue integrity following Cγ treatment. To generate an insulin resistance model, male Wistar rats were provided 30% sucrose solution ad libitum for 20 weeks. To generate a type 2 diabetes model (STZ), five-day-old rats were intraperitoneally injected with streptozotocin (150 mg/kg). Each animal model was randomized into three subgroups that received the following oral treatments daily for one week: 0.9% w/v NaCl (vehicle; IR-Ctrl and STZ-Ctrl); metformin 300 mg/kg (IR-Met and STZ-Met); and Cγ 150 mg/kg (IR-Cγ and STZ-Cγ). Biochemical parameters were assessed pre- and post-treatment using colorimetric or enzymatic methods. We also performed histological analysis of hepatic and renal tissue. G6pc, Fbp1, and Pck1 gene expressions were quantified using real-time PCR. No histological changes were observed in any group. Post-treatment G6pc gene expression was decreased in the IR-Cγ and STZ-Cγ groups. Post-treatment Fbp1 and Pck1 gene expressions were reduced in the IR-Cγ group but increased in STZ-Cγ animals. Overall, these findings suggest that Cγ is involved in reducing hepatic glucose production, mainly through G6pc inhibition in impaired glucose metabolism disorders.     

Prediction of parental combination for introduction of stay-green associated loci in wheat

In wheat, the increase of yield and stability associated traits can be achieved by combining parents containing the stay-green trait and favorable alleles for grain yield. The aim of this work was to analyze the genetic dissimilarity between wheat lines from stay-green and synchronized maturation groups and elite cultivars. Moreover, to propose promising combinations seeking the selection of high-grain yield and high bread-making quality genotypes containing stay-green trait. The experiment was conducted in a randomized block design with three replications in 2003, 2004, and 2005, using sister-lines with the presence and absence of stay-green trait and elite cultivars. Genetic variability exists among wheat strains from the synchronized stay-green maturation group and elite cultivars. Genotypes of maturation group stay-green obtained an average performance superior to the synchronized group. Crosses between stay-green lines and the CEP 29 and BRS 177 cultivars are promising in the selection of genotypes carrying the stay-green trait with high yield and bread-making quality.