Effect of diet on phosphine toxicity, rate of development and reproduction of the rice weevil Sitophilus oryzae (Linnaeus)

One of the loci responsible for strong phosphine resistance encodes dihydrolipoamide dehydrogenase (DLD). The strong co-incidence of enzyme complexes that contain DLD, and enzymes that require thiamine as a cofactor, motivated us to test whether the thiamine deficiency of polished white rice could influence the efficacy of phosphine fumigation against insect pests of stored grain. Three strains of Sitophilus oryzae (susceptible, weak and strong resistance) were cultured on white rice (thiamine deficient), brown rice or whole wheat. As thiamine is an essential nutrient, we firstly evaluated the effect of white rice on developmental rate and fecundity and found that both were detrimentally affected by this diet. The mean time to reach adult stage for the three strains ranged from 40 to 43 days on brown rice and 50–52 days on white rice. The mean number of offspring for the three strains ranged from 7.7 to 10.3 per female over a three day period on brown rice and 2.1 to 2.6 on white rice. Growth and reproduction on wheat was similar to that on brown rice except that the strongly resistant strain showed a tendency toward reduced fecundity on wheat. The susceptible strain exhibited a modest increase in tolerance to phosphine on white rice as expected if thiamine deficiency could mimic the effect of the dld resistance mutation at the rph2 locus. The strongly resistant strain did not respond to thiamine deficiency, but this was expected as these insects are already strongly resistant. We failed, however, to observe the expected synergistic increase in resistance due to combining thiamine deficiency with the weakly resistant strain. The lack of interaction between thiamine content of the diet and the resistance genotype in determining the phosphine resistance phenotype suggests that the mode of inhibition of the complexes is a critical determinant of resistance.

Optimizing Western Flower Thrips Management on French Beans by Combined Use of Beneficials and Imidacloprid

Western flower thrips (WFT), Frankliniella occidentalis (Pergande), is an important pest of vegetable crops worldwide and has developed resistance to many insecticides. The predatory mites Neoseiulus (=Amblyseius) cucumeris (Oudemans), the entomopathogenic fungus Metarhizium anisopliae (Metsch.), and an insecticide (imidacloprid) were tested for their efficacy to reduce WFT population density and damage to French bean (Phaseolus vulgaris L.) pods under field conditions in two planting periods. Metarhizium anisopliae was applied as a foliar spray weekly at a rate of one litre spray volume per plot while imidacloprid was applied as a soil drench every two weeks at a rate of two litres of a mixture of water and imidacloprid per m(2). Neoseiulus cucumeris was released every two weeks on plant foliage at a rate of three mites per plant. Single and combined treatment applications reduced WFT population density by at least three times and WFT damage to French bean pods by at least 1.7 times compared with untreated plots. The benefit-cost ratios in management of WFT were profitable with highest returns realized on imidacloprid treated plots. The results indicate that M. anisopliae, N. cucumeris, and imidacloprid have the potential for use in developing an integrated pest management program against WFT on French beans.

SUSCEPTIBILITY TO BRUCHIDS AMONG COMMON BEANS IN UGANDA

The bean bruchids, Acanthoscelides obtectus Say and Zabrotes subfasciatus Boheman (Coleoptera: Bruchidae), are cosmopolitan pests of stored dry common beans ( Phaseolus vulgaris L. ), causing damage through reduction of grain quality and seed germination. Biological resistance to these bruchids was definitively established in non-cultivated bean accessions, and has been introgressed into a range of drybean market classes. However, existing resistance to bruchids in Uganda’s common bean germplasm has not been systematically studied. In this study, 45 bean genotypes from the National Bean-Breeding Programme (25 genotypes) and agroecologically diverse bean growing areas in Uganda (20 genotypes), were evaluated for postharvest bruchid resistance. None of the evaluated bean genotypes expressed resistance to either bruchid species, with all the 45 bean genotypes supporting bruchid development, reproduction and feeding. All genotypes were severely damaged by bruchids feeding, resulting in significant (P<0.05) reduction of seed germination. Reduction in seed germination was related to the number of emergence holes and seed size; small bean seeds damaged by up to 2 bruchid emergence holes had a 7.1% reduction in germination, while large bean seeds with a similar number of emergence holes showed a 25% reduction in germination. Whereas this study further confirms bruchids as important storage pests of beans causing direct loss through consumption of the seed and indirect loss through viability deterioration, the resistance to bruchids in the evaluated range of Uganda’s dry bean germplasm is inadequate for direct exploitation in a breeding programme.

EFFECT OF TEMPORARY DROUGHT AT DIFFERENT GROWTH STAGES ON SNAP BEAN POD QUALITY AND YIELD

High quality snap bean ( Phaseolus vulgaris L. ) can be produced under rain-fed conditions, provided that adequate moisture is available. However, drought may occur at any stage of growth of snap bean. The objective of this study was to evaluate the effect of drought stress at different growth stages on pod physical quality and nutrient concentrations. An experiment was conducted at the Horticulture Greenhouse, Hawassa University in Ethiopia. Drought stress (50% of field capacity [FC]) was applied at the unfolding of the fourth trifoliate leaf, flowering and pod formation, against a control with no drought stress. The drought stress treatments and eight cultivars were arranged as a factorial experiment in a completely randomised design, with three replications. Drought stress (50% FC) during reproductive stages significantly (P<0.05) reduced pod texture, appearance, and pod curvature. Drought stress increased protein and zinc concentrations by 41 and 15%, respectively; but reduced iron concentration by 15% in snap bean pods. All the tested cultivars had relatively similar responses to drought stress.

Avaliação da ação bioinseticida de SBTI e vicilina de Erythrina velutina em enzimas digestivas e membrana peritrófica de larvas de Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests

NUTRITIONAL AND AMINO ACID ANALYSIS OF RAW, PARTIALLY FERMENTED AND COMPLETELY FERMENTED LOCUST BEAN ( Parkia biglobosa ) SEEDS

The nutritional and amino acid analysis of raw and fermented seeds of Parkia biglobosa were carried out. Parameters investigated include moisture, crude protein, crude fat, ash, crude fibre and mineral contents; and the effect of the degree of fermentation on these parameters was also investigated. The amino acid compositions of all the samples were evaluated and amino acid quality determined by calculating amino acid scores and the predicted protein efficiency ratio (P-PER). Results showed that the proximate composition was significantly affected by fermentation, although there was little difference between the parameters for the partially fermented and completely fermented samples. Based on dry matter percentage, protein content was in the 39.77 – 43.74 % range while crude fibre ranged between 5.55 – 7.42 %. The ash content was lowest in the raw sample (2.34 %), while the fermented samples had ash contents between 4.27 and 8.33 % for the fully fermented and the partially fermented seeds, respectively. The fat content increased from 8.65 % in the raw seed to 24.4 % and 27.6 % for the partially and completely fermented samples, respectively. Results of the amino acid analysis showed that the partially fermented sample had the lowest quantities of all amino acids determined and had lysine as the limiting amino acid, whereas the raw and completely fermented samples had very similar amino acid profile with amino acid scores of 100, indicating that there are no limiting amino acids. All the samples were rich in essential amino acids. The P-PER also showed that the partially fermented sample had the lowest protein efficiency while the raw seed had the highest. Mineral contents generally increased from the raw, through the partially fermented, to the completely fermented seeds and results showed the samples to be good sources of potassium (K), calcium (Ca), manganese (Mn) and copper (Cu) in addition to being complementary sources of other metals. Locust bean seed does not accumulate lead and is, therefore, safe for consumption without the potential of food poisoning.

Development of Locust Bean Gum Nanoparticles for protein delivery

Dissertação de Mestrado, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2014

Inhalable antitubercular therapy mediated by locust bean gum microparticles

Tuberculosis remains a major global health problem and alternative therapeutic approaches are needed. Considering the high prevalence of lung tuberculosis (80% of cases), the pulmonary delivery of antitubercular drugs in a carrier system capable of reaching the alveoli, being recognised and phagocytosed by alveolar macrophages (mycobacterium hosts), would be a significant improvement to current oral drug regimens. Locust bean gum (LBG) is a polysaccharide composed of galactose and mannose residues, which may favour specific recognition by macrophages and potentiate phagocytosis. LBG microparticles produced by spray-drying are reported herein for the first time, incorporating either isoniazid or rifabutin, first-line antitubercular drugs (association efficiencies >82%). Microparticles have adequate theoretical properties for deep lung delivery (aerodynamic diameters between 1.15 and 1.67 μm). The cytotoxic evaluation in lung epithelial cells (A549 cells) and macrophages (THP-1 cells) revealed a toxic effect from rifabutin-loaded microparticles at the highest concentrations, but we may consider that these were very high comparing with in vivo conditions. LBG microparticles further evidenced strong ability to be captured by macrophages (percentage of phagocytosis >94%). Overall, the obtained data indicated the potential of the proposed system for tuberculosis therapy.

Integrated pest management and the (un)sustainable use of pesticides

Doutoramento em Engenharia Agronómica - Instituto Superior de Agronomia - UL

Physical and chemical properties of starch and flour from different common bean (Phaseolus vulgaris L.) cultivar.

The physical, chemical and pasting properties of the flour and isolated starches from six different bean cultivars (Phaseolus vulgaris L.) were investigated in order to obtain information for application in new products. The protein and total starch contents of the bean flours ranged from 17.72 to 20.27% and from 39.68 to 43.78%, respectively. The bean starches had low amounts of proteins, lipids and ash and showed an amylose content ranging between 45.32 and 51.11% and absolute density values between 1.55 and 1.78 g.cm?3. The bean starch granules were round to oval with a smooth surface. Results viscoamylographic profiles of the starches and flours showed the possibility of selecting cultivars for specific applications according to these characteristics.

Drought impact on rainfed common bean production areas in Brazil.

Common bean production in Goiás, Brazil is concentrated in the same geographic area, but spread acrossthree distinct growing seasons, namely, wet, dry and winter. In the wet and dry seasons, common beansare grown under rainfed conditions, whereas the winter sowing is fully irrigated. The conventional breed-ing program performs all varietal selection stages solely in the winter season, with rainfed environmentsbeing incorporated in the breeding scheme only through the multi environment trials (METs) wherebasically only yield is recorded. As yield is the result of many interacting processes, it is challengingto determine the events (abiotic or biotic) associated with yield reduction in the rainfed environments(wet and dry seasons). To improve our understanding of rainfed dry bean production so as to produceinformation that can assist breeders in their efforts to develop stress-tolerant, high-yielding germplasm,we characterized environments by integrating weather, soil, crop and management factors using cropsimulation models. Crop simulations based on two commonly grown cultivars (Pérola and BRS Radi-ante) and statistical analyses of simulated yield suggest that both rainfed seasons, wet and dry, can bedivided in two groups of environments: highly favorable environment and favorable environment. Forthe wet and dry seasons, the highly favorable environment represents 44% and 58% of production area,respectively. Across all rainfed environment groups, terminal and/or reproductive drought stress occursin roughly one fourth of the seasons (23.9% for Pérola and 24.7% for Radiante), with drought being mostlimiting in the favorable environment group in the dry TPE. Based on our results, we argue that eventhough drought-tailoring might not be warranted, the common bean breeding program should adapttheir selection practices to the range of stresses occurring in the rainfed TPEs to select genotypes moresuitable for these environments.

Major loci controlling resistance to the angular leaf spot of common bean.

Based on the evidences presented in this paper, results from classical genetic studies, fine-mapping information and physical position analysis using the reference genome sequence of P. vulgaris, the BIC Genetic Committee has formally accepted the proposed new gene symbols.

Resistance to Fusarium wilt in common bean cultivars and lines in pre-commercial stage.

The objective of this study was to identify common bean cultivars with resistance to Fusarium wilt.

Phenotypic and molecular characterization of common bean genotypes for resistance to anthracnose.

This study aimed to perform phenotypic and molecular characterization of cultivars and breeding lines of common bean for resistance to anthracnose.

Cooking time of common bean: genetic parameters and selection of elite lines.

The objectives of this study were to investigate the genetic variability and select elite lines for CT, since these lines aggregate essential agronomic traits.