Biological and molecular properties of Tomato rugose mosaic virus (ToRMV), a new tomato-infecting begomovirus from Brazil

A viral complex causing golden mosaic and leaf distortion (rugosity) in tomato plants was obtained from viruliferous whiteflies, and named TGV-Ub1. This complex was sap-transmitted from tomato to Nicotiana benthamiana . PCR amplification using universal begomovirus primers yielded two distinct fragments for DNA-A, suggesting that the TGV-Ub1 complex comprised at least two distinct viruses. Clones corresponding to full-length viral genomes were obtained from tomato plants infected with TGV-Ub1. Comparisons of the complete sequences of clones pUb1-49 (DNA-A), pUb1-62 and pUb1-81 (both DNA-B) indicated that they constitute novel western hemisphere begomoviruses. Clones pUb1-49 and pUB1-81 have identical common regions, thus representing the cognate DNA-A and -B of a novel begomovirus, named Tomato rugose mosaic virus (ToRMV). Clone pUb1-62 has a distinct common region from ToRMV and all other geminiviruses. A cognate DNA-A for pUb1-62 was not found. Clones containing 1·8 copies of the genomic components were constructed. Infectivity assays of these clones in tomato and N. benthamiana demonstrated that the clones corresponding to ToRMV systemically infected both hosts. Symptoms were analogous to those observed when using the pure isolates obtained in this study. The combination of pUb1-49 and -62 did not result in systemic infection, indicating that these components do not form a viable virus. ToRMV was sap-transmitted from N. benthamiana to N. benthamiana , and by grafting to Solanum tuberosum and Datura stramonium . ToRMV-A and ToRMV-B were detected in plants of Nicandra physaloides and Phaseolus vulgaris , respectively, growing in nearby tomato fields, in association with distinct DNA components.     

Weather-based prediction of anthracnose severity using artificial neural network models

Data were collected and analysed from seven field sites in Australia, Brazil and Colombia on weather conditions and the severity of anthracnose disease of the tropical pasture legume Stylosanthes scabra caused by Colletotrichum gloeosporioides . Disease severity and weather data were analysed using artificial neural network (ANN) models developed using data from some or all field sites in Australia and/or South America to predict severity at other sites. Three series of models were developed using different weather summaries. Of these, ANN models with weather for the day of disease assessment and the previous 24 h period had the highest prediction success, and models trained on data from all sites within one continent correctly predicted disease severity in the other continent on more than 75% of days; the overall prediction error was 21·9% for the Australian and 22·1% for the South American model. Of the six cross-continent ANN models trained on pooled data for five sites from two continents to predict severity for the remaining sixth site, the model developed without data from Planaltina in Brazil was the most accurate, with >85% prediction success, and the model without Carimagua in Colombia was the least accurate, with only 54% success. In common with multiple regression models, moisture-related variables such as rain, leaf surface wetness and variables that influence moisture availability such as radiation and wind on the day of disease severity assessment or the day before assessment were the most important weather variables in all ANN models. A set of weights from the ANN models was used to calculate the overall risk of anthracnose for the various sites. Sites with high and low anthracnose risk are present in both continents, and weather conditions at centres of diversity in Brazil and Colombia do not appear to be more conducive than conditions in Australia to serious anthracnose development.     

A meta-analysis on experimental infections with porcine circovirus type 2 (PCV2)

A meta-analysis was performed with the aim to identify factors with a relevant influence on the expression of clinical postweaning multisystemic wasting syndrome (PMWS) under experimental conditions. Data from 44 studies were included in the analysis. Several variables were studied: number of pigs in the experiment, intake of colostrum, serological status against porcine circovirus type 2 (PCV2), strain of PCV2 used for inoculation, the route and dose of inoculation, and use of potential triggering factors (such as co-infections, vaccinations, or immunomodulator products). Multiple correspondence analysis and log-linear regression methods were used to establish the relationships between the studied variables and the number of PCV2 infected pigs that developed PMWS. Based on the results of the meta-analysis, the most successful animal experiment aimed to develop PMWS should include: (1) colostrum-deprived pigs, (2) age of inoculation below 3 weeks, (3) high doses of PCV2 inoculum, (4) PCV2 strain from genotype 1, and (5) co-infection with another swine pathogen as a triggering factor.     

Digestive peptidases and proteinases in the midgut gland of the pink shrimp Farfantepenaeus paulensis (Crustacea, Decapoda, Penaeidae)

Proteases from the midgut gland of the Farfantepenaeus paulensis juveniles were assessed. Enzyme activity was determined using protease substrates and inhibitors. The effect of pH, temperature and calcium on proteolytic activity was assayed. Caseinolytic activity was analysed in substrate-sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Trypsin, chymotrypsin and leucine aminopeptidase activity was detected. Proteolytic activity was strongly inhibited by the specific trypsin inhibitors. Tosyl-phenylalanine chloromethyl ketone inhibited 59.3% of chymotrypsin activity. The greatest trypsin-like activity occurred at pH 8.0 and 45 °C. Chymotrypsin-like activity reached maximal values at alkaline pH (7.2–9.0) and 55 °C. CaCl₂ did not increase trypsin-like activity, but rather inhibited it at concentrations of 30 (20%), 50 (30%) and 100 mM (50%). The substrate-SDS-PAGE zymogram revealed eight proteinase bands. Two possibly thermal-resistant (85 °C, 30 min) chymotrypsin isoforms were found, which were inhibited by phenyl-methyl-sulphonyl-fluoride. Aminopeptidase activity of enzyme extracts (Arg, Leu, Lys, Phe and Val) and the recommended concentrations of these essential amino acids in penaeid shrimp diets were positively correlated ( P <0.05). Beause protein digestion involves the combined action of different enzymes, adequate knowledge of shrimp digestion and enzyme characteristics is required for the assessment of the digestive potential of different feed sources and development of in vitro digestibility protocols.     

Does the Use of Chitosan Contribute to Oxalate Kidney Stone Formation?

Chitosan is widely used in the biomedical field due its chemical and pharmacological properties. However, intake of chitosan results in renal tissue accumulation of chitosan and promotes an increase in calcium excretion. On the other hand, the effect of chitosan on the formation of calcium oxalate crystals (CaOx) has not been described. In this work, we evaluated the antioxidant capacity of chitosan and its interference in the formation of CaOx crystals in vitro. Here, the chitosan obtained commercially had its identity confirmed by nuclear magnetic resonance and infrared spectroscopy. In several tests, this chitosan showed low or no antioxidant activity. However, it also showed excellent copper-chelating activity. In vitro, chitosan acted as an inducer mainly of monohydrate CaOx crystal formation, which is more prevalent in patients with urolithiasis. We also observed that chitosan modifies the morphology and size of these crystals, as well as changes the surface charge of the crystals, making them even more positive, which can facilitate the interaction of these crystals with renal cells. Chitosan greatly influences the formation of crystals in vitro, and in vivo analyses should be conducted to assess the risk of using chitosan.     

Lead accumulation and its effects on the branchial physiology of Prochilodus lineatus

The purpose of this work was to determine the tissue accumulation of lead (Pb) and its effects on osmoregulatory processes of the freshwater fish Prochilodus lineatus. Juvenile fish were exposed to Pb (from 1.7 to 0.7 mg of dissolved Pb L^?1) for 6, 24 and 96 h and Pb accumulation was analyzed in the gills, liver, kidneys, blood cells and muscle. The following parameters were also analyzed: hematologic (hemoglobin content, hematocrit and number of red blood cells), metabolic (blood glucose), endocrine (blood cortisol), osmo ionic (plasma osmolality and Na⁺, K⁺, Cl^? and Ca⁺² concentrations), gill enzymes (Na⁺/K⁺-ATPase and carbonic anhydrase), chloride cell (CC) density and CC location in the gills. Pb accumulated in all the analyzed tissues, with the kidneys showing the highest concentration, followed by the gills and liver. The lowest Pb concentrations were found in blood cells and muscle. Pb promoted an increase in blood glucose after 6 and 24 h exposure. Gill Na⁺/K⁺-ATPase was inhibited after 24 h of exposure, but its activity was restored at 96 h, probably due to the increase in CC in gill lamellae. Plasma Na⁺ was reduced after 6 and 96 h, while K⁺ concentrations increased at all the experimental times. Fish exposed to Pb showed reduced plasma Ca⁺² at all experimental periods. Hematologic parameters remained unchanged. Overall, this study demonstrated that Pb interferes in osmoregulatory processes of P. lineatus and the proliferation of CC in the gills is a response in order to reestablish adequate ion concentrations.     

Establishing the optimal essential amino acid ratios in juveniles of Nile tilapia (Oreochromis niloticus) by the deletion method

One of the strategies used to improve fish production is lowering the feed costs and the environmental impact by reducing dietary protein content. Using the deletion method, we determined the optimal amino acid (AA) ratio for Nile tilapia (Oreochromis niloticus) (body weight 20 g). Eleven experimental diets and four replications, taken two at a time, distributed in a randomized block design were used. For this trial, a balanced diet (BD) was formulated. Ten other diets were formulated by the deletion method, in which the BD diet was adjusted to result in a reduction of 45% of the test amino acid. Fish were fed three times a day for 57 days. Groups of fish at the beginning and at the end of the experiment were euthanized for further determination of the carcass nitrogen (N) composition. The optimal ratio of each AA was derived by dividing the requirement for each AA by the requirements for lysine. The essential AA ratios, expressed relative to lysine (=100), were methionine 64, threonine 93, tryptophan 24, arginine 125, histidine 34, isoleucine 57, leucine 96, valine 76 and phenylalanine 101. Our findings might be used to design strategies aimed at reducing the production costs of Oreochromis niloticus.     

A framework for results‐based management in fisheries

We present a framework for results‐based management (RBM) of commercial fisheries. The core idea of RBM is to reduce micromanagement by delegating management responsibility to resource users. The RBM framework represents an industrial organization approach to co‐management and comprises three defining processes, conducted by three independent “agents”: (i) an “authority” defines specific and measurable and achievable objectives (outcome targets, OTs) for the utilization of fisheries resources, (ii) resource user organizations (termed “operators”) take responsibility for achieving these OTs and provide documentation that (iii) allows independent “auditors” to evaluate the achievement of OTs. Using incentive mechanisms, notably deregulation, RBM grants operators the flexibility to develop and implement innovative and cost‐effective ways to achieve OTs. The feasibility of implementing RBM in five European fisheries was investigated in cooperation with relevant stakeholders through artificial planning processes and computer simulations. The operators involved were enthusiastic, and new management plans were drafted based on the framework. These included socioeconomic OTs in addition to traditional stock objectives, encompassing an ecosystem approach. Several issues are in need of further research to consolidate the approach and prepare the ground for practical implementation, including: the specification of the legal and regulatory framework required to underpin RBM, details of transitional arrangements when shifting towards RBM (including cost‐sharing) and the development of necessary organizational capacity for operators. Initially, we therefore envisage the framework being applied to high‐value single‐species fisheries, with a limited number of participants, which are adequately represented by a competent organization.