Application of Six Detection Methods for Analysis of Paralytic Shellfish Toxins in Shellfish from Four Regions within Latin America

With the move away from use of mouse bioassay (MBA) to test bivalve mollusc shellfish for paralytic shellfish poisoning (PSP) toxins, countries around the world are having to adopt non-animal-based alternatives that fulfil ethical and legal requirements. Various assays have been developed which have been subjected to single-laboratory and multi-laboratory validation studies, gaining acceptance as official methods of analysis and approval for use in some countries as official control testing methods. The majority of validation studies conducted to date do not, however, incorporate shellfish species sourced from Latin America. Consequently, this study sought to investigate the performance of five alternative PSP testing methods together with the MBA, comparing the PSP toxin data generated both qualitatively and quantitatively. The methods included a receptor binding assay (RBA), two liquid chromatography with fluorescence detection (LC-FLD) methods including both pre-column and post-column oxidation, liquid chromatography with tandem mass spectrometry (LC-MS/MS) and a commercial lateral flow assay (LFA) from Scotia. A total of three hundred and forty-nine shellfish samples from Argentina, Mexico, Chile and Uruguay were assessed. For the majority of samples, qualitative results compared well between methods. Good statistical correlations were demonstrated between the majority of quantitative results, with a notably excellent correlation between the current EU reference method using pre-column oxidation LC-FLD and LC-MS/MS. The LFA showed great potential for qualitative determination of PSP toxins, although the findings of high numbers of false-positive results and two false negatives highlighted that some caution is still needed when interpreting results. This study demonstrated that effective replacement methods are available for countries that no longer wish to use the MBA, but highlighted the importance of comparing toxin data from the replacement method using local shellfish species of concern before implementing new methods in official control testing programs.     

Soil bulk density and porosity of homogeneous morphological units identified by the Cropping Profile Method in clayey Oxisols in Brazil

Soil structure is important to root development and crop yield. The objective of this study was to test the Cropping Profile Method in Brazilian soils, in order to evaluate the soil structure in the field. Grouped different structures determined by the Cropping Profile Method were compared to laboratory determinations for soil bulk density, total porosity and mercury porosity. The study was conducted in clayey Oxisols submitted to different uses and management including annual crops, orchards and natural forests in the State of Paraná, southern Brazil. Homogeneous morphological units (HMUs) were determined in trenches using the Cropping Profile Method, and the different structures were grouped as: (a) non-compacted; (b) compacted; (c) in-process-of-compacting. Results of field evaluation were compatible with those obtained in the laboratory. More compacted and in-process-of-compacting structures corresponded to soil bulk density values of 1.42 and 1.33 Mg m⁻³, which were significantly higher than the 1.18 Mg m⁻³ value obtained for soil bulk density in non-compacted HMU. The total porosity of compacted HMU and in-process-of-compacting HMU was 0.49 and 0.52 m³ m⁻³, respectively. These were significantly lower than the value obtained for the non-compacted HMU (0.60 m³ m⁻³). The Cropping Profile Method is useful mainly in field research works when it is important to verify the effect of management practices on soil structure.     

Physicochemical properties of natural phenolics from grapes and olive oil byproducts and their antioxidant activity in frozen horse mackerel fillets

The reducing and chelating capacities and the affinity for the incorporation into the fish muscle of grape procyanidins, hydroxytyrosol, and propyl gallate were studied together with their antioxidant activity in frozen horse mackerel (Trauchurus trauchurus) fillets. Fillets were supplemented with phenolic antioxidants by (a) spraying an aqueous phenolic solution, (b) glazing with an aqueous phenolic solution, and (c) a previous washing of fillets with water plus spraying an aqueous phenolic solution. The effect of washing on the endogenous pro-oxidant/antioxidant balance of the fillets was also determined. All phenolic compounds were effective delaying lipid oxidation in the fish fillets. The order of antioxidant efficiency in spraying and glazing was propyl gallate > hydroxytyrosol > procyanidins, which was similar to the reducing power of these phenolics, but did not show any correlation with their chelating capacity and their affinity to the fish muscle. Washing the fillets with water prior to spraying phenols increased synergistically the antioxidant activity of grape procyanidins and changed the relative antioxidant efficiency to propyl gallate approximately procyanidins > hydroxytyrosol. This synergism may be a result of a better distribution of the procyanidins onto the fillet surface because of the residual water that remained on the fillets surface after washing.     

Selectivity of diacylhydrazine insecticides to the predatory bug Orius laevigatus: in vivo and modelling/docking experiments

BACKGROUND: Knowledge of pesticide selectivity to natural enemies is necessary for a successful implementation of biological and chemical control methods in integrated pest management (IPM) programmes. Diacylhydrazine (DAH)‐based ecdysone agonists, also known as moulting‐accelerating compounds (MACs), are considered to be a selective group of insecticides, and their compatibility with predatory Heteroptera, which are used as biological control agents, is known. However, their molecular mode of action has not been explored in beneficial insects such as Orius laevigatus (Fieber) (Hemiptera: Anthocoridae). RESULTS: In this project, in vivo toxicity assays demonstrated that the DAH‐based RH‐5849, tebufenozide and methoxyfenozide have no toxic effect against O. laevigatus. The ligand‐binding domain (LBD) of the ecdysone receptor (EcR) of O. laevigatus was sequenced, and a homology protein model was constructed that confirmed a cavity structure with 12 α‐helices, harbouring the natural insect moulting hormone 20‐hydroxyecdysone. However, docking studies showed that a steric clash occurred for the DAH‐based insecticides owing to a restricted extent of the ligand‐binding cavity of the EcR of O. laevigatus. CONCLUSIONS: The insect toxicity assays demonstrated that MACs are selective for O. laevigatus. The modelling/docking experiments are indications that these pesticides do not bind with the LBD‐EcR of O. laevigatus and support the supposition that they show no biological effects in the predatory bug. These data help in explaining the compatible use of MACs together with predatory bugs in IPM programmes. Copyright © 2012 Society of Chemical Industry     

Physiometabolic responses to road transport in weaned piglets for a short period and the effects of straw bedding

The aim of the study was to evaluate the physiometabolic responses to stress in hybrid suckling piglets that were weaned and transported at different ages on a paved road under two vehicle conditions: with and without a layer of straw bedding. The piglets were weaned and transported at the following ages: 8, 15 and 22 days. The duration of each trip was 1 h. The piglets in each group were further divided into two sub‐groups for two distinct experimental transport conditions: (1) over the road with straw bedding; and (2) over the same road but without straw bedding. A total of three repetitions were performed for each experimental condition. In order to assess their responses to weaning‐ and transport‐induced stress, the study evaluated a thorough physiometabolic blood profile. The 8‐ and 15‐day‐old piglets transported without straw bedding showed increases in their pCO2, blood glucose, blood lactate levels and the percentage of hematocrit, but a decrease of pO2 upon arrival (P < 0.05). In contrast, the 22‐day‐old piglets transported on the same road with straw bedding were able to re‐establish their blood concentrations of lactate, pCO2, pO2, Na+, K+, Ca2+ and pH during the trip (P < 0.05).     

A synthetic conjugate polysaccharide vaccine against Haemophilus influenzae type b

Glycoconjugate vaccines provide effective prophylaxis against bacterial infections. To date, however, no commercial vaccine has been available in which the key carbohydrate antigens are produced synthetically. We describe the large-scale synthesis, pharmaceutical development, and clinical evaluation of a conjugate vaccine composed of a synthetic capsular polysaccharide antigen of Haemophilus influenzae type b (Hib). The vaccine was evaluated in clinical trials in Cuba and showed long-term protective antibody titers that compared favorably to licensed products prepared with the Hib polysaccharide extracted from bacteria. This demonstrates that access to synthetic complex carbohydrate-based vaccines is feasible and provides a basis for further development of similar approaches for other human pathogens.     

Gluconic acid consumption in wines by Schizosaccharomyces pombe and its effect on the concentrations of major volatile compounds and polyols

Schizosaccharomyces pombe 1379 (ATCC 26760) yeast strain in wine substantially increases acetaldehyde and 1,1-diethoxyethane concentrations and to decreases levo-2,3-butanediol, glycerol, acetoin, and gluconic acid concentrations. In this study, S. pombe has been used for the first time to reduce gluconic acid in wine under aerobic conditions. Only acetaldehyde and acetoin exhibited significantly higher levels in the wines containing gluconic acid. The high in vitro specific activity of alcohol dehydrogenase observed may be directly related to the high production of acetaldehyde by the studied fission yeast.     

Antimicrobial activity of olive solutions from stored Alpeorujo against plant pathogenic microorganisms

The aim of this work was to assess the in vitro antimicrobial effects that wastewaters from alpeorujo oil extraction have against phytopathogenic bacteria and fungi. Alpeorujo was stored for 6 months and then processed to extract its oil, pomace, and a new liquid waste (OWSA), which was characterized by its content in phenolic compounds. OWSA at 20% decreased bu >4 log the population of Erwinia spp., Pseudomonas spp., and Clavibacter spp. viable cells in test tubes, whereas OWSA at 50% in agar medium was necessary to inhibit mycelial growth of most fungi. It was found that the bactericidal effect was due to the joint action of low molecular mass phenolic compounds, although neither hydroxytyrosol, its glucosides, hydroxytyrosol glycol, nor a glutaraldehyde-like compound individually explained this bioactivity. Hence, OWSA constitutes a promising natural solution to fight plant phytopathogenic bacteria and fungi.