BTH-mediated antioxidant system responses in apple leaf tissues

BTH (S-methylbenzo-1,2,3-thiadiazole-7-carbothiate), an active compound of the commercial preparation Bion, has been studied as an elicitor of resistance to fire blight (Erwinia amylovora) in apple. However, the biochemical mechanisms of its action are not fully elucidated. Our study indicated that BTH at the best time of its protection activity (2–14 days after application) induced changes in prooxidant–antioxidant balance in the leaves of apple trees, but in different ways in the enzymatic and nonenzymatic antioxidants. Glutathione as low molecular antioxidant as well as superoxide anion radical and lipid peroxides as oxidants exhibited changes at the early phase of BTH action. Glutathione-dependent enzymes were strongly affected by the elicitor used. On the 2nd day glutathione transferase (GST) and glutathione peroxidase (GSH-Px) activities increased by about 70% and 30% above the control, respectively. GST activity normalized about the 14th day but GSH-Px at the same time showed 27% of the control value. Among enzymes utilising hydrogen peroxide only catalase showed increase (37%) at the early phase of experiment. Compared with the control, BTH-treated plants did not show changes in ascorbate peroxidase and phenylalanine ammonia-lyase activities. Tocopherol (TOC) level diminished starting from the 7th day after BTH treatment and on the 14th day it was only 28% of the control. It is proposed that extinguishing of BTH-mediated signal resulted from TOC and glutathione action. The diminished ascorbate level at all examined times may play a crucial role in BTH-mediated cell growth regulation. The direct influence of BTH on lipid metabolism should be also taken into consideration.     

New qualitative approach in the characterization of antioxidants in white wines by antioxidant free radical scavenging and NMR techniques

The aim of this study was to obtain new information on antioxidant compounds in white wines. For this purpose, white wine degradation was promoted by a forced aged protocol, and six normally aged white wines from different vintages were analyzed. Both normal and forced aged wines were sequentially extracted using hexane and ethyl acetate. Apolar antioxidants were removed using hexane, and polar antioxidants were extracted with ethyl acetate. This last residue was subject to partial re-extraction with hexane and acetone. The antioxidant capacity of the wines and of each fraction was evaluated by two free radical methods, ABTS and DPPH. Normal aging provides a decrease in the total antioxidant capacity of wines. The antioxidant activity of ethyl acetate/acetone extracts was approximately 95% higher than that found for the hexane extracts. Concerning the forced aged wines, results showed that the wine submitted to a temperature of 60 degrees C for 21 days had higher antioxidant activity than that submitted to a temperature of 20 degrees C. With regard to the ethyl acetate/acetone extracts, oxygen and temperature treatment leads to a decrease in their antioxidant activity. NMR analysis was performed in the highest antioxidant capacity organic fractions (ethyl acetate/acetone extracts) and in the aqueous fraction of the control wine (T = 20 degrees C), in order to attempt the characterization of species involved in oxygen protection. Possible structures of antioxidant compounds in white wines were proposed. Two of these are tyrosol-like structures. This molecule is a well-known phenolic compound in wine, and it is reported to have antioxidative effects.     

Enhancement of the rootability of a structurally fragile tropical soil using gypsum and leguminous residues to increase the yield of maize

In humid tropical areas, a combination of factors related to soil conditions for root development contribute to decreased nutrient use efficiency. Our aim was to evaluate the combined effects of the application of gypsum and mulch from leguminous trees on rootability of root zone soil, root growth, phosphorus recovery efficiency and maize yield in a sandy loam soil prone to hardsetting. The experiment involved a randomized block design with four replications and six treatments: no residue or gypsum (control, C); leguminous residues (L); leguminous residues and 6 t/ha of gypsum (LG6); urea and 6 t/ha of gypsum (UG6); leguminous residues, urea and 6 t/ha of gypsum (LUG6); and leguminous residues, urea and 12 t/ha of gypsum (LUG12). Application of gypsum plus leguminous residues enhanced soil rootability, when applied to a tropical Arenic Hapludult, through the combined effects of increased porosity and decreased penetration resistance. These improvements facilitated root growth in deeper soil layers and greater nutrient uptake. Application of leguminous residues increased the input and retention of calcium in deeper layers. The use of large amounts of gypsum with leguminous residue, despite a significant increase in phosphorus uptake, did not result in a proportional increase in maize yield. Given the small differences in calcium content between the 6 and 12 t/ha gypsum treatments, applying a low dose of gypsum combined with leguminous residue, rather than administering a single large gypsum dose, is recommended for tropical sandy loam soils with low cation retention capacities.     

Treatment of 46 cats with porcine lente insulin--a prospective, multicentre study

This prospective, multicentre, non-blinded, open study followed 46 cats with diabetes mellitus during treatment with porcine lente insulin (also known as porcine insulin zinc suspension, Caninsulin, Intervet) for 16+/-1 weeks (stabilization phase), with additional monitoring of some cats (n=23) for a variable period. At least three of the following were present at initial presentation: appropriate history of clinical signs consistent with diabetes mellitus, glucosuria, blood glucose greater than 15 mmol/l and fructosamine greater than 380 micromol/l. Insulin treatment was started at a dose rate of 0.25-0.5 IU/kg body weight twice daily, with a maximum starting dose of 2 IU/injection. Twenty-eight of the cats were classed as reaching clinical stability during the study, in 23 of these cats this was during the stabilization phase. Seven cats went into remission during the stabilization phase and one of the cats in week 56. Clinical signs of hypoglycaemia, significantly associated with a dose of 3 units or 0.5 IU/kg or more per cat (twice daily), were observed in nine of the 46 cats during the stabilization phase and concomitant biochemical hypoglycaemia was recorded in most cases. Biochemical hypoglycaemia, recorded in 6% of the blood glucose curves performed during the stabilization phase, was significantly associated with a dose rate of 0.75 IU/kg or more twice daily. This further highlights the need for cautious stepwise changes in insulin dose. The protocol used in the present study is suitable for and easy to use in practice. This study confirmed the efficacy and safety of porcine lente insulin (Caninsulin) in diabetic cats under field conditions.     

Genetic alterations of KIT during clonal expansion and subsequent acquisition of resistance to toceranib in a canine mast cell tumor cell line

One of the potential mechanisms underlying acquired resistance to toceranib in canine mast cell tumor (MCT) is the emergence of a secondary mutation in the KIT gene. Here, genetic alterations of KIT during clonal expansion and subsequent acquisition of resistance to toceranib were investigated in the toceranib‐susceptible canine MCT cell line VI‐MC, which carries a KIT‐activating mutation resulting in a predicted p.(Asn508Ile) amino acid change in the receptor tyrosine kinase protein KIT. Two sublines were cloned from VI‐MC and toceranib‐resistant sublines then were established by continuous exposure to toceranib. The mutation status of KIT in parental VI‐MC and its sublines was investigated using next‐generation sequencing (NGS). Additionally, effects of secondary mutations on toceranib sensitivity in p.(Asn508Ile)‐mutant KIT were examined. KIT secondary mutations, including those encoding p.(Asn679Lys)‐, p.(Asp819Val)‐, and p.(Asp819Gly)‐mutant KIT, that confer toceranib insensitivity to p.(Asn508Ile)‐mutant KIT emerged only in toceranib‐resistant VI‐MCs. These mutations were not detected by NGS in the parental VI‐MC line or in the toceranib‐naive cloned VI‐MCs, although the parental line and sublines exhibited genetic heterogeneity in KIT that may have been caused by genetic evolution during clonal expansion. VI‐MC clones with these secondary mutations in KIT appear to have arisen from subclones during treatment with toceranib rather than being pre‐existing. However, further study using a higher resolution technique will be needed to confirm the developmental mechanism of KIT secondary mutation in canine MCT cells with acquired resistance to toceranib.     

Comparative foraging behaviour of three species of shrike in southern Africa

Utilisation of food resources in migrant and resident birds living sympatrically often leads to differences in feeding strategy. For sit-and-wait hunters such as shrikes, it is connected with competition for food and hunting places. In this study, foraging behaviour of three shrike species was studied in savanna-like habitat in southern Africa. The fieldwork was carried out in December 2017 during wintering season of Lesser Grey Shrike Lanius minor and Red-backed Shrike Lanius collurio and breeding season of Common Fiscal Lanius collaris. Details of the bird's behaviour were recorded during 30 min observations of each bird. A general linear mixed model with logit link function and binomial error variance was used to compare the species’ behaviour. Although the type of perch-site and perching time did not differ between species, Red-backed Shrike used places situated significantly lower than other shrikes. This species also more often used bushes and attacked from lower perch-sites than Lesser Grey Shrike. This may possibly be due to interspecific competition and antipredator behaviour of Red-backed Shrike. No differences in foraging behaviour between Common Fiscal and Lesser Grey Shrike were observed. All three species utilised the same category of prey. Hunting success (the ratio of successful to all attacks) was low and similar in all three species (47%–57%). The lack of statistically significant differences in behaviour, hunting success and size of prey items among shrike species may be a consequence of the relatively short observation time and small sample sizes.     

Identification of the causal agents of crazy root disease on hydroponically cultivated cucumber plants in Poland

European Journal of Plant Pathology - In April 2019, hydroponically cultivated cucumber plants with characteristic symptoms of crazy root disease were found in two different commercial production...     

Diversity of Botryosphaeriaceae causing grapevine trunk diseases and their spatial distribution under different climatic conditions in Algeria

The family Botryosphaeriaceae is one of the most widespread and cosmopolitan endophytic group of fungi. Every year, species of this family cause severe damages on table and wine grape production, worldwide. However, this threat is still poorly known in Algeria. In this study, a large number of Botryosphaeriaceae-like isolates were obtained from symptomatic grapevines collected from eight regions with different ecological conditions, namely: Boumerdès, Médéa, Algiers, Tipaza, El Taref, Sidi Bel Abbes, Biskra and Adrar. The isolates were identified using DNA sequences of the translation elongation factor (tef1-α) and internal transcribed spacer (ITS) regions. Eleven species belonging to six genera, including Neofusicoccum parvum, N. algeriense, N. vitifusiforme, N. stellenboschiana, N. luteum, Diplodia seriata, D. olivarum, Lasiodiplodia theobromae, Dothiorella sarmentorum, Botryosphaeria dothidea and Neoscytalidium dimidiatum were identified. The spatial distribution of the Botryosphaeriaceae showed that D. seriata and N. stellenboschiana were the most widespread in the Algerian vineyards, while L. theobromae was recorded in the desert region of Biskra. Pathogenicity trials showed that all species were pathogenic on detached green shoots of grapevine, with N. parvum and L. theobromae being the most aggressive.

     

Immunological properties of recombinant classical swine fever virus NS3 protein in vitro and in vivo

Classical swine fever (CSF) is a highly contagious and often fatal disease of pigs characterised by fever, severe leukopenia and haemorrhages. With vaccines having an importance in disease control, studies are seeking improved protein-based subunit vaccine against the virus (CSFV). In this respect, recombinant viral NS3 protein was analysed for its immunopotentiating capacity, particularly in terms of cytotoxic immune responses. NS3 was effective at inducing in vitro responses, quantified by lymphoproliferation, IFN-gamma ELISPOT, flow cytometric detection of activated T cell subsets, and cytotoxic T cell assays. Peripheral blood mononuclear cells from CSFV-immune pigs could be stimulated, but not cells from na?ve animals. In addition to the IFN-gamma responses, induction of both CD4+ T helper cell and CD8+ cytotoxic T cells (CTL) were discernible--activation of the latter was confirmed in a virus-specific cytolytic assay. Attempts were made to translate this to the in vivo situation, by vaccinating pigs with an E2/NS3-based vaccine compared with an E2 subunit vaccine. Both vaccines were similar in their abilities to stimulate specific immune responses and protect pigs against lethal CSFV infection. Although the E2/NS3 vaccine appeared to have an advantage in terms of antibody induction, this was not statistically significant when group studies were performed. It was also difficult to visualise the NS3 capacity to promote T-cell responses in vivo. These results show that NS3 has potential for promoting cytotoxic defences, but the formulation of the vaccine requires optimisation for ensuring that NS3 is correctly delivered to antigen presenting cells for efficient activation of CTL.     

Antifungal activity evaluation of the constituents of Haliclona baeri and Haliclona cymaeformis, collected from the Gulf of Thailand

A new compound maleimide-5-oxime was isolated, together with 3,4-dihydroxybenzoic acid, tetillapyrone, from the ethyl acetate extract of the marine sponge Haliclona baeri while tetillapyrone, nortetillapyrone, p-hydroxybenzaldehyde and phenylacetic acid were isolated from the ethyl acetate extract of Haliclona cymaeformis, collected from the Gulf of Thailand. The structures of tetillapyrone and nortetillapyrone were re-examined using HMBC correlations. Maleimide-5-oxime, tetillapyrone and nortetillapyrone were found to be inactive against three human tumor cell lines (the estrogen-dependent ER(+) MCF-7, the estrogen-independent ER(−) MDA-MB-231 and NCI-H460. Maleimide-5-oxime, p-hydroxybenzaldehyde, phenylacetic acid, tetillapyrone and nortetillapyrone were evaluated for their growth inhibitory effect against seven yeasts and eight filamentous fungi. Only nortetillapyrone showed antifungal activity, with a preponderance on the dermatophytic filamentous fungi.