The Role of Fumonisins in the Biological Interaction between Fusarium verticillioides and Sitophilus zeamais.

The aim of the current study was to investigate the entomopathogenic capacity of the mold Fusarium verticillioides and the effect of its mycotoxins fumonisins, on the grain beetle Sitophilus zeamais. We evaluated the capacity of this fungus to infect live insects, the antifungal activity of constituents of the insect's epicuticle, and the effect of a fumonisin extract on the fitness of the insects. We found that F. verticillioides could not penetrate the cuticle of S. zeamais and that the fumonisin extract had no negative effects on the fitness of the insects. However, the progeny of the insects increased, and the fumonisin extract had repellent effects. This is the first report about the effects of fumonisins on the relationship between F. verticillioides and S. zeamais, which may provide useful information about interactions between pathogenic microorganisms and insects, especially on stored product pests.

Mycoviruses in Biological Control: From Basic Research to Field Implementation.

Mycoviruses from plant pathogens can induce hypovirulence (reduced virulence) in their host fungi and have gained considerable attention as potential biocontrol tools. An increasing number of mycoviruses that induce fungal hypovirulence, from a wide variety of taxonomic groups, are currently being reported. Successful application of these viruses in disease management is greatly dependent on their ability to spread in the natural populations of the pathogen. Mycoviruses generally lack extracellular routes of transmission. Hyphal anastomosis is the main route of horizontal mycovirus transmission to other isolates, and conidia of vertical transmission to the progeny. Transmission efficiencies are influenced by both the fungal host and the infecting virus. Interestingly, artificial transfection methods have shown that potential biocontrol mycoviruses often have the ability to infect a variety of fungi. This expands their possible use to the control of pathogens others than those where they were identified. Mycovirus research is also focused on gaining insights into their complex molecular biology and the molecular bases of fungus-virus interactions. This knowledge could be exploited to manipulate the mycovirus and/or the host and generate combinations with enhanced properties in biological control. Finally, when exploring the use of mycoviruses in field conditions, the pathogen life style and the characteristics of the disease and crops affected will deeply impact the specific challenges to overcome, and the development of biocontrol formulations and delivery methods.

In vitro activity of natural phenolic compounds against fluconazole-resistant Candida species: a quantitative structure-activity relationship analysis.

AIMS: To evaluate the antifungal activity and to analyse the structure-activity relationship of eleven natural phenolic compounds against four Candida species which are resistant to fluconazole. METHODS AND RESULTS: Four different species of Candida isolates were used: Candida albicans, Candida krusei, Candida tropicalis and Candida dubliniensis. The phenolic compound carvacrol showed the highest anti-Candida bioactivity, followed by thymol and isoeugenol. The obtained minimum inhibitory concentration (MIC) values obtained were used in a quantitative structure-activity relationship (QSAR) analysis where the electronic, steric, thermodynamic and topological descriptors served as dependent variables. According to the descriptors obtained in this QSAR study, the antifungal activity of phenols has a first action specific character which is based on their interaction with plasma or mitochondrial membranes. The second action is based on a steric descriptor-the maximal and minimal projection of the area-which could explain the inability of some phenolic compounds to be biotransformed to quinones methylene by Candida species. CONCLUSIONS: According to the descriptors obtained in this QSAR study, the anti-Candida activity of ortho-substituted phenols is due to more than one action mechanism. The anti-Candida activity of phenolic compounds can be predicted by their molecular properties and structural characteristics. SIGNIFICANCE AND IMPACT OF THE STUDY: These results could be employed to predict the anti-Candida activity of new phenolic compounds in the search for new alternatives or complementary therapies to combat against candidiasis.

Effects of thymol and isoeugenol feed supplementation on quail adult performance, egg characteristics and hatching success.

1. A study was conducted to evaluate whether feed supplementation with thymol or isoeugenol can alter Japanese quail growth rate and final body weight, the female onset of puberty, hen-day egg production and the physical and chemical characteristics of the egg, as well as its potential to alter hatchability. 2. From 4 to 16 weeks of age, birds from each cage (1 male: 3 females) were assigned to 1 of 3 treatments that differed in the supplement added to the feed: control, thymol or isoeugenol (400 mg/kg). The average ages (d) at first egg lay (FIRST), at 25% egg production (A25% EP), at 50% egg production (A50% EP) and weekly and cumulative hen-day egg production (HDEP) were calculated. In addition, physical and chemical characteristics of the eggs, their fertility and hatchability were also evaluated for each group. 3. Feed supplementation did not significantly affect growth rate, final body weight, egg production parameters, fertility and physical characteristics of egg or most of the fatty acid components of the yolk. 4. The group treated with isoeugenol showed an increase in the percentage of palmitoleic fatty acid compared to the control, with thymol group showing intermediates values. 5. Both thymol and isoeugenol supplemented groups showed increased hatchabilities, by 18.8% and 11.8%, respectively, compared to their control counterparts. 6. The improvement in the hatching success of the eggs from the thymol and isoeugenol supplemented groups without a negative impact on their performance may have important economic implications for future breeding programmes, particularly if these effects generalise from quail to other more commercially important poultry species, such as chickens or turkeys.

Inhibitory effect of cyclic terpenes (limonene, menthol, menthone and thymol) on Fusarium verticillioides MRC 826 growth and fumonisin B1 biosynthesis.

The minimum inhibitory concentration (MIC) of cyclic terpenes (limonene, menthol, menthone and thymol) against Fusarium verticillioides MRC 826 was assessed by using the semisolid agar antifungal susceptibility (SAAS) technique. Limonene, menthol, menthone and thymol were evaluated at final concentrations of 25, 50, 75, 150, 200, 250, 500 and 1000 microL/L of culture medium. Limonene and thymol showed the highest inhibitory effects on F. verticillioides development. Thus, the effects of monoterpenes on fumonisin B1 (FB1) biosynthesis were evaluated by using corn grain (Zea mays) as substrate. The monoterpenes were inserted on maize 1 day before inoculation with a conidial suspension of F. verticillioides to give final concentrations of 75 ppm. At this concentration, thymol was the most active inhibitor on FB1 biosynthesis.

Thymol as natural antioxidant additive for poultry feed: oxidative stability improvement.

Antioxidant are regularly included in poultry feed as protection from deterioration during storage. Recently the interest for the use of natural phytochemicals in animal diets has been increased. Thymol (THY) has been proven to be an effective antioxidant for extending broiler meat quality during storage with similar action to the widely used butylated hydroxytoluene (BHT). This study evaluates whether THY can also have a protective effect on the feed mash by assessing its antioxidant potential and related changes in fatty acid (FA) balance. Feed mash was assigned to 1 of 4 treatments, control (CON, no additive), vehicle (VEH, ethanol 96%), BHT (400 mg BHT /kg feed) and THY (400 mg THY /kg feed). Three replicates of each treatment were taken after 0, 30, and 60 d of storage at room temperature (23 ± 3°C) and relative humidity (40 ± 5%). Peroxide value (PV), titratable acidity (TA) and FA relative composition were determined. As expected, there were no treatment effects on those variables at 0 d of storage. However, higher PV values were detected in the CON and VEH groups after 30 and 60 d of storage in comparison to the THY and BHT treated samples (CON = VEH > THY = BHT). While a slight increase was also observed in TA through storage time, no particular treatment effects were detected. Relative FA composition changed with storage time only in the CON and VEH group which had a decrease in polyunsaturated fatty acids and an increase in saturated FA. No changes were detected in the Thy and BHT treated feeds. The results suggest a similar THY and BHT protective effect on feed mash lipid oxidation. Thus, THY could be considered as a useful natural alternative to help sustain quality of poultry feed.

Subchronic mycotoxicoses in Wistar rats: assessment of the in vivo and in vitro genotoxicity induced by fumonisins and aflatoxin B(1), and oxidative stress biomarkers status.

Some evidence suggests that fumonisin B(1) (FB(1)), a worldwide toxic contaminant of grains produced by Fusarium verticillioides, exhibits an oxidative stress mediated genotoxicity. We studied the DNA damage (by the alkaline comet and the micronucleus tests) and biomarkers of cellular oxidative stress (malondialdehyde, MDA; catalase, CAT; and superoxide dismutase, SOD) in spleen mononuclear cells of male Wistar rats subchronically (90 days) fed on a control experimental diet (CED) or poisoned with experimental diets contaminated with a culture material containing 100 ppm of FB(1) (FED), with 40 ppb of aflatoxin B(1) (a common toxic co-contaminant in cereals, AFB(1)ED), and with a mixture of both toxins (MED). The DNA damage was found in 13.7%, 81.7%, 98.0% and 99.3% (comet assay) and in 2.8%, 7.0%, 10.8% and 8.8% (micronucleus technique) in groups CED, FED, AFB(1)ED and MED, respectively. The MDA levels as well as the CAT and SOD activities were increased in all the poisoned animals. A similar behavior was observed in cells exposed in vitro to the toxins. These data support the hypothesis of an oxidative stress mediated genotoxicity induced by FB(1). Furthermore, the extent of DNA damage assessed by the comet assay suggests a possible protective effect of the fumonisins-AFB(1) mixtures in vitro against the genotoxicity induced individually by the toxins.