Correlation of molecular markers and biological properties in Verticillium dahliae and the possible origins of some isolates

Haploid and amphihaploid Verticillium dahliae isolates were studied using PCR-based molecular markers which: (i) discriminate the defoliating and nondefoliating pathotypes (two primer pairs INTD2f/r and INTND2f/r), and (ii) are species-specific (primer pair 19/22). The results were compared with some known biological and other molecular properties of the isolates. Five discrete sequences of the 19/22 amplicon were found. Sequence 4 was associated with both defoliating isolates from Spain and nondefoliating isolates from Spain and USA; these pathotypes were separated by the primer pairs INTD2f/r and INTND2f/r, but the data showed that the primer espdef01 (derived from the 19/22 amplicon) cannot be used for this purpose. Amplicon sizes and sequences with primers 19/22 divided amphihaploid isolates from crucifers (thought to be interspecific hybrids) into those corresponding to the previously reported α and β groups. The β-group isolates had either sequence 4 or 5 (these two differing by a single base). The distinct amplicon sequence 3 given by the α-group isolates demonstrated that the V. dahliae -like 'parent' of this group was molecularly unlike any haploid isolate yet studied. The overall results are discussed in relation to phytosanitary considerations and the probability of defoliating or crucifer pathotypes arising de novo within Europe, either by selection or by interspecific hybridizations.     

Antimicrobial and Antibiofilm Effect of Inulin-Type Fructans,Used in Synbiotic Combination with Lactobacillus spp. Against Candida albicans

Plant Foods for Human Nutrition - There is great interest in the search for new alternatives to antimicrobial drugs, and the use of prebiotics and probiotics is a promising approach to this...     

The role of small reserves in plant conservation in a region of high diversity in eastern Spain

The Valencian region, in eastern Spain, is home to ≈3050 vascular plant species, 68 of which are listed under the critically endangered IUCN category. To afford protection to our endangered, rare and endemic flora, a network of small (2-20 ha) statutory reserves has been created by the Regional Wildlife Service. These are termed “micro-reserves” and encompass a large number of individual species and natural habitats. The objective is to monitor long term changes in the plant populations and to carry out active management of the protected plants. In the selection of plots, criteria of distinctiveness and endangerment of plants was followed. Eight years after the onset of this initiative, plant micro-reserves have become an essential tool for the effective protection of the diverse flora of this Mediterranean region.     

Influence of S. babylonica extract on feed intake, growth performance and diet in vitro gas production profile in young lambs

An experiment was completed to determine the effect of Salix babylonica (SB) extract supplementation to the diet of growing lambs. Eighteen Katahdin × Pelibuey male lambs (14?±?2 kg live body weight) were divided randomly in individual cages into three groups and fed three diets varying in SB: a control group was fed on total mixed ration (TMR) without SB (SB0), an SB25 group was fed on TMR plus SB extract at 25 mL/lamb/day, and an SB50 group was fed on TMR plus SB extract at 50 mL/lamb/day on dry matter intake (DMI), average daily gain (ADG), feed efficiency, and in vitro gas production (GP) in lambs fed on TMR. In vitro GP of the TMR fed to lambs was recorded at 2, 4, 6, 8, 10, 12, 24, 48, and 72 h of incubation with 0, 0.6, 1.2, and 1.8 mL extract per gram of DM. Addition of SB extract at low and high doses improved the DMI of lambs by 59.9 and 33.2 %, respectively. Relative to the control, low and high extract doses achieved greater lamb ADG during the experimental period. The asymptotic GP increased (P?<?0.05) with increasing dose of SB extract without affecting the rate of GP or the initial delay before GP begins. Linear increases for in vitro GP with advancing time with different SB extract doses were observed. It is suggested that the use of S. babylonica extract with the rate of 25 mL/lamb/day is beneficial to young lamb’s performance growth and thus can be safely used as a feed additive in diets without any negative effects on animal health.     

Variation of pathotypes and races and their correlations with clonal lineages in Verticillium dahliae

Understanding pathogenic variation in plant pathogen populations is key for the development and use of host resistance for managing verticillium wilt diseases. A highly virulent defoliating (D) pathotype in Verticillium dahliae has previously been shown to occur only in one clonal lineage (lineage 1A). By contrast, no clear association has yet been shown for race 1 with clonal lineages. Race 1 carries the effector gene Ave1 and is avirulent on hosts that carry resistance gene Ve1 or its homologues. The hypothesis tested was that race 1 arose once in a single clonal lineage, which might be expected if V. dahliae acquired Ave1 by horizontal gene transfer from plants, as hypothesized previously. In a diverse sample of 195 V. dahliae isolates from nine clonal lineages, all race 1 isolates were present only in lineage 2A. Conversely, all lineage 2A isolates displayed the race 1 phenotype. Moreover, 900‐bp nucleotide sequences from Ave1 were identical among 27 lineage 2A isolates and identical to sequences from other V. dahliae race 1 isolates in GenBank. The finding of race 1 in a single clonal lineage, with identical Ave1 sequences, is consistent with the hypothesis that race 1 arose once in V. dahliae. Molecular markers and virulence assays also confirmed the well‐established finding that the D pathotype is found only in lineage 1A. Pathogenicity assays indicated that cotton and olive isolates of the D pathotype (lineage 1A) were highly virulent on cotton and olive, but had low virulence on tomato.     

Undernutrition in the parental and first generation provokes an organ‐specific response to oxidative stress on neonates of second filial generation of Wistar rats

Undernutrition induces an increase of the oxidative stress that can predispose offspring to various diseases in adulthood through epigenetic reprogramming. The aim of this study was to evaluate the effects of intergenerational undernutrition on protein oxidation and antioxidant defence response on liver, heart and brain of the second‐generation neonates (F₂) of undernourished rats. For this purpose, both parents in parental (F₀) and first generation (F₁) were fed with a low‐nutrient diet. Body mass and length decreased (p < 0.05) in F₀, F₁ and F₂ being the F₁ males who exhibited a greater mass loss. A decrease in plasma albumin concentration was observed in F₂ neonates (p < 0.05) and also a mass loss of liver, heart and brain (p < 0.05), although proportionally to body length reduction. Undernutrition increased levels of protein oxidation in liver and heart (p < 0.05) but not in brain (p > 0.05) while catalase activity increased only in brain (p < 0.05). In summary, intergenerational undernutrition modifies the antioxidant status through an organ‐specific response, on F₂ neonate rats, where the brain increased catalase activity to prevent a severe oxidative damage and support the vital functions of this key organ to maintain vital functions.     

Diversity of insecticide resistance mechanisms and spectrum in European populations of the codling moth, Cydia pomonella

Only a few of the registered insecticides against Cydia pomonella L. are still effective in areas where insecticide resistance has emerged in this pest. Resistance mechanisms are multiple, and their lone or cumulative effects in a single population are not completely understood. A detailed estimation of resistance spectrum is still required to define the suitable insecticides to use against a given population. The efficacy of ten insecticides was therefore investigated together with the resistance mechanisms expressed in four laboratory strains and 47 field populations of C. pomonella from five countries. Bioassays were performed using topical applications of diagnostic concentrations on diapausing larvae, and resistance mechanisms were analysed on adults emerging from control insects. All populations exhibited a reduced susceptibility to at least one insecticide when compared with the susceptible laboratory strain. Cross-resistances were observed between azinphos-methyl or phosalone and more recent compounds such as spinosad and thiacloprid. Resistances to azinphos-methyl, diflubenzuron, spinosad, tebufenozide and thiacloprid were significantly correlated with mixed-function oxidase activity, while increased glutathione-S-transferase and reduced non-specific esterase activities were correlated with resistance to azinphos-methyl and emamectin, respectively. Conversely, resistances to azinphos-methyl, tebufenozide and thiacloprid were negatively correlated with increased esterase activity. None of the observed mechanisms explained the loss of susceptibility of populations to chlorpyrifos-ethyl, and no significant correlation was detected between resistance to deltamethrin and the presence of the kdr mutation. The suitability of such non-target instars to monitor insecticide resistance in field populations is discussed.     

Effect of Mediterranean saltbush (Atriplex halimus) ensilaging with two developed enzyme cocktails on feed intake,nutrient digestibility and ruminal fermentation in sheep

The aim of this study was to assess the effects of feeding Atriplex halimus (AH) silage treated with two developed enzyme cocktails to sheep on feed intake, nutrient digestibility and ruminal fermentation. The AH silage was treated without or with 2 L of ZAD1^® or ZAD2^®/1000 kg with 5% molasses and ensiled for 30 days. Barley grain (300 g/head/day) was fed as an energy supplement once daily at 10.00 hours and AH silage with or without enzyme treatment was offered ad libitum to animals twice daily at 09.00 and 16.00 hours. Sheep were fed on four experimental forage diets comprised of AH silage and barley (D1), AH silage treated with ZAD1^® and barley (D2), AH silage treated with ZAD2^® and barley (D3) and AH silage treated with a combination of ZAD1^® and ZAD2^® (1:1) and barley (D4). Ensiling AH with enzymes reduced its contents of neutral detergent fiber and acid detergent fiber. The dry matter intake of AH of D2, D3 and D4 decreased (P < 0.001) as compared to D1. However, enzyme‐treated diets had greater total digestible nutrients intake (P < 0.001) as compared to D1. The nutrients digestibility for D2, D3 and D4 were higher than those for D1 (P < 0.001), and were higher for D3 as compared to both D2 and D4. Sheep fed on D3 had highest (P < 0.001) ruminal total volatile fatty acids concentration, ammonia nitrogen concentration and microbial protein yield. It could be concluded that AH silage treated with ZAD1^® or ZAD2^® improved digestibility and rumen fermentation in sheep.     

Nutritional reference values for Opuntia ficus‐indica determined by means of the boundary‐line approach

Optimum concentrations and/or sufficiency ranges of nutrients are useful for a correct diagnosis and improvement of nutrient status of cultivated plants. To develop boundary‐line approach (BLA) standards for Opuntia ficus‐indica L., a database of N, P, K, Ca, and Mg concentrations in 1‐year‐old cladodes and cladodes fresh‐matter yield was used. The BLA optimum concentrations (associated with estimated maximum yield) for O. ficus‐indica were: N = 13.1 g kg^–1, P = 3.2 g kg^–1, K = 44.4 g kg^–1, Ca = 38.1 g kg^–1, and Mg = 17.3 g kg^–1. The BLA sufficiency ranges at 95% yield were 8.4–20.3 g kg^–1 for N, 2.4–4.2 g kg^–1 for P, 38.2–50.8 g kg^–1 K, 31.8–45.2 g kg^–1 for Ca, and 14.3–20.9 g kg^–1 for Mg. The BLA standards are comparable to those obtained in a previous study using compositional‐nutrient‐diagnosis (CND) approach.