Identification of genetic sources with attenuated <Emphasis Type="Italic">Tomato chlorosis virus</Emphasis>-induced symptoms in <Emphasis Type="Italic">Solanum</Emphasis> (section <Emphasis Type="Italic">Lycopersicon</Emphasis>) germplasm

The whitefly-transmitted Tomato chlorosis virus (ToCV) (genus Crinivirus) is associated with yield and quality losses in field and greenhouse-grown tomatoes (Solanum lycopersicum) in South America. Therefore, the search for sources of ToCV resistance/tolerance is a major breeding priority for this region. A germplasm of 33 Solanum (Lycopersicon) accessions (comprising cultivated and wild species) was evaluated for ToCV reaction in multi-year assays conducted under natural and experimental whitefly vector exposure in Uruguay and Brazil. Reaction to ToCV was assessed employing a symptom severity scale and systemic virus infection was evaluated via RT-PCR and/or molecular hybridization assays. A subgroup of accessions was also evaluated for whitefly reaction in two free-choice bioassays carried out in Uruguay (with Trialeurodes vaporariorum) and Brazil (with Bemisia tabaci Middle-East-Asia-Minor1—MEAM1?=?biotype B). The most stable sources of ToCV tolerance were identified in Solanum habrochaites PI 127827 (mild symptoms and low viral titers) and S. lycopersicum ‘LT05’ (mild symptoms but with high viral titers). These two accessions were efficiently colonized by both whitefly species, thus excluding the potential involvement of vector-resistance mechanisms. Other promising breeding sources were Solanum peruvianum (sensu lato) ‘CGO 6711’ (mild symptoms and low virus titers), Solanum chilense LA1967 (mild symptoms, but with high levels of B. tabaci MEAM1 oviposition) and Solanum pennellii LA0716 (intermediate symptoms and low level of B. tabaci MEAM1 oviposition). Additional studies are necessary to elucidate the genetic basis of the tolerance/resistance identified in this set of Solanum (Lycopersicon) accessions.     

Combining ability of common bean parents in different seasons,locations and generations

The objectives of this study were to obtain information about genetic parameters of a partial diallel in common bean, as well as to evaluate the effects of growing seasons, locations and generations in these estimates in order to define the best strategies for the use of diallel analysis as methodology of selection of parents and populations in common bean breeding. Twelve parents were crossed in a partial diallel scheme (5?×?7), and the F₁ and F₂ generations were evaluated in four experiments (I, II, III, and IV). The effects of seasons were determined in the Experiments I and II, in which the F₁ generation was evaluated in two distinct growing seasons. To quantify the effect of locations, we used Experiments III and IV, in which the F₂ generation was evaluated in the same season, but at different locations, and to quantify the effect of generations, the experiments II (generation F₁) and III (F₂ generation) were evaluated in the same season and location. As for grain yield the genetic control varied according to the environment. An effect of interaction on the genetic parameters of the diallel was observed for generation, whereas for grain yield, effects of season and location were observed. For plant architecture, parental combining ability should therefore be estimated using the F₁ generation, while for grain yield, both the F₁ and F₂ can be used. The recombination between the best hybrids is a promising strategy for extraction of superior black bean lines.     

Dietary Protein Hydrolysates from Animal By-Products: Digestibility and Enzymatic Activity for Dourado Salminus brasiliensis

Fishmeal (FM) is a high-cost and scarce feedstuff. Notwithstanding, the formulation and processing of diets for carnivorous fish strongly rely on FM as a protein source, given its palatability and biological value. Animal by-product hydrolysates are high-quality feedstuff and can substitute FM in aquafeeds. This study evaluated the digestibility of hydrolysates from tilapia residue (TR), tuna head (TH), swine liver (SL), and poultry liver (PL), and the profile of digestive enzymes in juvenile dourado, Salminus brasiliensis, a carnivorous Characin, fed diets containing graded levels of the hydrolysates. Inclusion of hydrolysates in diet formulations lowered the pH of feed but did not alter feed intake by fish. Higher apparent digestibility coefficients (ADCs) of diets were recorded for fish fed diets containing TR and SL, and lower ADCs were recorded for fish fed diets containing TH. Protease and lipase activities in the animal’s stomach were higher, especially for those fed with diets containing SL. Amylase activity was higher in pyloric caeca, whereas in the intestines, the higher activity was seen in fish fed control and TR diets. Animal by-product hydrolysates were highly digestible for dourado, and the enzymatic profile of fish was influenced by the nutrient contents of diets.     

Depletion study and estimation of withdrawal periods for florfenicol and florfenicol amine in pacu (Piaractus mesopotamicus)

The intensive production of farmed fish is at a global all‐time high, and the control of bacteria proliferation in fish farms requires the frequent use of antimicrobials. This practice raises important environmental concerns related to the emergence of antimicrobial‐resistant bacteria strains. Only a few antimicrobial drugs have been approved for use in aquaculture, one of which is florfenicol. This work studies the depletion and withdrawal period of florfenicol and its main metabolite, florfenicol amine, in pacu (Piaractus mesopotamicus), a neotropical characin widely farmed in the southern hemisphere. Juvenile pacu (average weight of 724 g) were stocked in a closed‐loop laboratory system with controlled water temperature (25.8°C), and were fed for 10 consecutive days with a diet containing an intended dose of 10 mg/florfenicol per kg bw. Muscle and skin tissues were collected at 1, 3, 6, 8, 10, 12 and 16 days post‐treatment, and florfenicol and florfenicol amine were quantified using a validated ultra‐high performance liquid chromatography‐tandem mass spectrometry (UPLC‐MS/MS) method. The limits of quantitation for florfenicol and florfenicol amine were 10 ng/g in muscle and 50 ng/g in skin. Considering a maximum residue limit of 1000 ng/g for the sum of florfenicol and florfenicol amine in muscle with skin in natural proportions a withdrawal period of 5 days (water temperature 25.8°C) or 129 degree days was calculated on the basis of the upper limit of the one‐sided 95% confidence interval for the 99^th percentile derived from the residue depletion study.     

Could the Supply of Boron and Zinc Improve Resistance of Potato to Early Blight?

The adequate supply of boron (B) and zinc (Zn) can improve the structural integrity and permeability of cell membranes, which is a defence mechanism against fungi infection. A greenhouse experiment was conducted to investigate the effect of the application of B and Zn on plant growth and the incidence and severity of potato early blight, a destructive foliar disease of potato crop caused by Alternaria grandis E.G. Simmons. Potato plants (Solanum tuberosum L. cv. Ágata) were grown in 11-l pots filled with sandy soil, unfertilized or fertilized with 5 mg kg^?1 B and 5 mg kg^?1 Zn (either alone or in combination). Potato plants were inoculated with isolates of Alternaria grandis at 40 days after planting. Early blight incidence and severity were evaluated visually 7 days after inoculation with A. grandis isolates. Disease incidence ranged from 16 to 41% infected leaves, and was significantly highest in the treatment with the application of B, followed by the control, and lowest with application of Zn and B + Zn. Early blight severity ranged from 2.5 to 25%, and was significantly higher in the unfertilized treatment than in those fertilized with Zn either alone or in combination with B. These findings suggest that Zn plays a critical role in potato tolerance to early blight and should be considered as a preventive measure in the disease management plan, since application of zinc reduced the incidence and severity of potato early blight. However, the physiological basis of this remains unknown.     

Adaptation and Validation of Colorimetric Methods in Determining Ammonium and Nitrate on Tropical Soils

Inorganic nitrogen (N) determination in soil extracts is a useful tool in studying soil–N interactions. The main method used worldwide is the steam distillation, but colorimetric methodologies show advantages of being quick, simple, sensitive, and higher yield. However, they are subject to disruption when the extract has interfering organic N, limiting its application in tropical soil. The aim of this study is to evaluate colorimetric methods for the determination of NH₄⁺ (through Berthelot reaction) and NO₃^? + NO₂^? (through Griess-Ilosvay reaction) in saline extracts of tropical soils by addition of activated charcoal during the extraction process. It is recommended to use extraction solution of 2 mol L^?1 potassium chloride (KCl) and reaction time of 30 min. Validation parameters confirmed the best performance of the colorimetric methods, revealing results favorable to the addition of the activated charcoal in the extraction process to increase the accuracy and precision of the colorimetric methods.     

Vitrification of sperm from marine fish: effect on motility and membrane integrity

Our goal was to develop a standardized approach for sperm vitrification of marine fish that can be applied generally in aquatic species. The objectives were to: (i) estimate acute toxicity of cryoprotectants over a range of concentrations; (ii) evaluate the properties of vitrification solutions (VS); (iii) evaluate different thawing solutions and (iv) evaluate sperm quality after thawing by examination of motility and membrane integrity. Sperm were collected from red snapper (Lutjanus campechanus), spotted seatrout (Cynoscion nebulosus) and red drum (Sciaenops ocellatus). A total of 29 combinations of cryoprotectants were evaluated for toxicity and glass formation. Samples were loaded onto 10‐μL polystyrene loops and plunged into liquid nitrogen. There was a significant difference (P < 0.05) in post‐thaw motility among VS and among species when using the same VS. The sperm in VS of 15% DMSO + 15% ethylene glycol + 10% glycerol + 1% X‐1000^? + 1% Z‐1000^? had an average post‐thaw motility of 58% and membrane integrity of 19% for spotted seatrout, 38% and 9% for red snapper, and 30% and 19% for red drum. Adaptations by marine fish to higher osmotic pressures could explain the survival in the high cryoprotectant concentrations. Vitrification offers an alternative to conventional cryopreservation.