Serenella Piano

Mechanisms of action and efficacy of four isolates of the yeast Metschnikowia pulcherrima active against postharvest pathogens on apples

Abstract The mechanisms of action and efficacy of four isolates (GS37, GS88, GA102, and BIO126) of the yeast Metschnikowia pulcherrima against Botrytis cinerea , Penicillium expansum , Alternaria sp., and Monilia sp., all postharvest pathogens of apple fruit, were studied in vitro and on apples, in controlled and semi-commercial conditions. An application of a cell suspension (10 8 cells per ml) of the antagonists in artificial wounds of apples reduced growth of B. cinerea and P. expansum after storage at 23xa0°C. A complete suppression of the pathogen was obtained against Monilia sp., stored at 23xa0°C, and against B. cinerea and P. expansum , stored at 4xa0°C. The results against Alternaria sp. were more variable. Applications of culture filtrates and autoclaved cells of the isolates were ineffective in reducing the diameter of the lesions on the fruit, supporting the hypothesis that living cells are necessary for biocontrol. In experiments of antagonism in vitro, on different solid substrates, a reduction in the mycelium growth of the pathogens resulted, so that, at least in vitro, the antagonists could produce some diffusible toxic metabolites. In co-cultivation in vitro on a synthetic medium, B. cinerea spore (10 5 ml −1 ) germination was completely inhibited by the presence of 10 8 cells of the antagonists, while culture filtrates and autoclaved suspensions were not able to reduce germination. Dipping boxes of apples cv. Golden delicious in a suspension of 10 7 antagonist cells per ml and storing for 8 months in controlled atmosphere at 1xa0°C, showed levels of control against B. cinerea and P. expansum similar to those from thiabendazole.

Biocontrol capability of Metschnikowia pulcherrima against Botrytis postharvest rot of apple

Abstract The biocontrol capability of two isolates of the yeast Metschnikowia pulcherrima against Botrytis postharvest rot of apple was studied in vitro and on apples under different storage conditions. The biocontrol activity of M. pulcherrima 2.33 or 4.4 can be reduced or totally suppressed by the addition of several nitrates. The addition of 100 g l−1 fructose inhibited the pathogen per se, while leaving unaltered the antagonistic capability of M. pulcherrima. These results support the hypothesis that competition for nutrients plays a major role in the biocontrol capability of M. pulcherrima against Botrytis postharvest rot of apple. However, both yeasts strongly inhibited the growth and spore germination of B. cinerea in vitro even under non-restrictive nutrient conditions. There was no evidence for the involvement of diffusible toxic metabolites in the biocontrol efficacy. The antagonistic activity of M. pulcherrima was dependent on the concentration of the antagonist. At 107–108 cells ml−1, 2.33 was most effective when applied at least 6 h before the pathogen, while 4.4 provided consistent results when co-inoculated with B. cinerea or applied 1 h later. Rapid colonisation of fresh apple fruit wounds was observed at room temperature during the first 24–48 h, and then the populations stabilised for the remaining storage period. On apple wounds kept at 4°C, the increase in population density of M. pulcherrima was lower, but continued over 96–128 h after application of the antagonists until it reached the same saturation level observed on apples stored at room temperature.

Clostridia spore formation during aerobic deterioration of maize and sorghum silages as influenced by Lactobacillus buchneri and Lactobacillus plantarum inoculants

Aims:u2002 The effect of the inoculation of maize and sorghum silages with Lactobacillus plantarum (LP) and Lactobacillus buchneri (LB) on the clostridia spore formation during aerobic deterioration has been studied.

Effect of Lactobacillus buchneri LN4637 and Lactobacillus buchneri LN40177 on the aerobic stability, fermentation products, and microbial populations of corn silage under farm conditions.

This study determined the efficacy of the use of 2 commercial inoculants containing Lactobacillus buchneri alone or in combination with homofermentative lactic acid bacteria in improving aerobic stability of corn silage stored in commercial farm silos in northern Italy. In the first survey, samples were collected from 10 farms that did not inoculate their silages and from 10 farms that applied a Pioneer 11A44 inoculant (L. buchneri strain LN4637; Pioneer Hi-Bred International, Des Moines, IA). In the second survey, corn silage samples were collected from 11 farms that did not inoculate their silages and from 11 farms that applied a Pioneer 11CFT inoculant (L. buchneri strain LN40177; Pioneer Hi-Bred International). Inoculants were applied directly through self-propelled forage harvesters, at the recommended rate of 1 g/t of fresh forage, to achieve a final application rate of 1.0 × 10(5) cfu/g of L. buchneri. One corn bunker silo, which had been open for at least 10 d, was examined in detail on each farm. The silages inoculated with L. buchneri had lower concentrations of lactic acid, a lower lactic-to-acetic acid ratio, a lower yeast count, and higher aerobic stability compared with the untreated silages. Unexpectedly, concentrations of acetic acid and 1,2-propanediol, 2 hallmarks of L. buchneri activity, did not differ between treatments and were only numerically higher in the inoculated silages compared with untreated ones, in both surveys. Aerobic stability, on average, was 107 and 121 h in the inoculated silages and 64 and 74 h in the untreated silages, for surveys 1 and 2, respectively, and decreased exponentially as the yeast count in the silage at the time of sampling increased, regardless of treatment. Inoculation with L. buchneri proved to be effective in reducing the yeast count to <2 log cfu/g of silage in 16 of 21 of the studied farm silages, confirming the ability of this inoculum to enhance the aerobic stability of corn silages in farm bunker silos.

Evolution of fungal populations in corn silage conserved under polyethylene or biodegradable films

To identify cultivable filamentous fungi before ensiling, after silage conservation, in farm‐silos covered with two different plastic films (polyethylene (PE) vs biodegradable (MB)), as well as after aerobic exposure of whole‐crop corn silage.

Leaching of S-metolachlor, terbuthylazine, desethyl-terbuthylazine, mesotrione, flufenacet, isoxaflutole, and diketonitrile in field lysimeters as affected by the time elapsed between spraying and first leaching event

The effect of elapsed time between spraying and first leaching event on the leaching behavior of five herbicides (terbuthylazine, S-metolachlor, mesotrione, flufenacet, and isoxaflutole) and two metabolites (desethyl-terbuthylazine and diketonitrile) was evaluated in a 2011–2012 study in northwest Italy. A battery of 12 lysimeters (8.4 m2 long with a depth of 1.8 m) were used in the study, each filled with silty-loam soil and treated during pre-emergence with the selected herbicides by applying a mixture of commercial products Lumax (4 L ha−1) and Merlin Gold (1 L ha−1). During treatment periods, no gravity water was present in lysimeters. Irrigation events capable of producing leaching (40 mm) were conducted on independent groups of three lysimeters on 1 day after treatment (1 DAT), 7 DAT, 14 DAT, and 28 DAT. The series was then repeated 14 days later. Leachate samples were collected a few days after irrigation; compounds were extracted by solid phase extraction and analyzed by high-performance liquid chromatography and gas chromatography–mass spectrometry. Under study conditions, terbuthylazine and S-metolachlor showed the highest leaching potentials. Specifically, S-metolachlor concentrations were always found above 0.25 µg L−1. Desethyl-terbuthylazine was often detected in leached waters, in most cases at concentrations above 0.1 µg L−1. Flufenacet leached only when irrigation occurred close to the time of herbicide spraying. Isoxaflutole and mesotrione were not measured (<0.1 µg L−1), while diketonitrile was detected in concentrations above 0.1 µg L−1 on 1 DAT in 2011 only.

Aerobic stability of maize silage stored under plastic films with different oxygen permeability

BACKGROUNDnThe most important factor that can influence silage quality is the degree of anaerobiosis maintained during conservation. The quality of the plastic film is a key factor, since the permeability of polyethylene to oxygen is too high for silage conservation. The aim of this work was to assess the effects of the interaction between three plastic films with different degrees of oxygen permeability and two different maize silage conservation times on fermentation, microbial quality and aerobic stability.nnnRESULTSnThe conservation time affected the pH, lactic and acetic acids, 1,2-propanediol and lactic/acetic acid ratio, with higher pH, lower lactic acid concentration and lactic/acetic acid ratio but higher acetic acid and 1,2-propanediol concentrations being found in silages conserved for 110 days versus silages conserved for 55 days. The plastic film affected the pH, lactic and acetic acids and lactic/acetic acid ratio. The yeast count was lower and aerobic stability higher for silage conserved under the oxygen barrier film for both conservation periods.nnnCONCLUSIONnThe positive interaction between length of storage and the increased anaerobiosis of silage during conservation provided by the oxygen barrier compared with polyethylene film helped reduce the yeast count and increase the aerobic stability of maize silage, even when less acetic acid was produced during ensiling.

Dissipation of Propanil and 3,4 Dichloroaniline in Three Different Rice Management Systems

This study focused on the dissipation of propanil and 3,4 dichloroaniline (3,4 DCA) over time in the soil, field water, inlet water, and outlet water of paddy fields under three management systems: conventional water seeding (CON), conventional water seeding with supplied liquid manure (LMA), and dry seeding (DRY). Propanil dissipation in water was also investigated under laboratory conditions. The field study was conducted from 2004 to 2006 at Vercelli, northern Italy. Propanil and 3,4 DCA showed rapid dissipation in water and soil environments both in the field and in the laboratory. Under controlled conditions, chemical hydrolysis was not detected for either compounds for up to 100 d at pHs of 5, 7, and 9. In the laboratory, the half-life of propanil in irrigation water was 1.1 d; its half-life in soil was routinely measured at <1.0 d (between 0.17 and 1.77 d). 3,4 DCA was found to persist much longer. Measured in all three study years at 50 d after treatment, its concentration ranged between 44 μg kg (CON) and 140 μg kg (DRY). Propanil and 3,4 DCA concentrations in paddy water were particularly high in samples collected at 4 d (2004) and 2 d (2005) after treatment. Maximum concentrations were 54.4 μg L (CON) for propanil (2005) and 113.7 μg L (LMA) for 3,4 DCA (2004). The concentrations of propanil and 3,4 DCA in inlet water were never above 1.1 and 0.3 μg L, respectively, whereas the highest concentration of each compound in outlet water was in samples collected first after treatment in 2005 and 2006. Both chemicals dissipated rapidly in all the soil-water environments but displayed no important differences among the three management systems. In conclusion, propanil and 3,4 DCA did not persist longer in paddy fields. A risk of water network contamination by these compounds may occur only early after herbicide spraying. A water-holding period after herbicide spraying may reduce this risk.

Buffer strip effect on terbuthylazine, desethyl-terbuthylazine and S-metolachlor runoff from maize fields in Northern Italy

The effectiveness of a 6 m wide vegetative buffer strip for reducing runoff of S-metolachlor, terbuthylazine and desethyl-terbuthylazine was studied in 2007–2008 in Northern Italy. Two cultivated fields, with and without the buffer strip, were compared. Residues of the chemicals were investigated in runoff water collected after runoff events and their dissipation in the soil was studied. The highest concentration of the chemicals in water occurred in samples collected from the unbuffered field at the first runoff events. Losses of terbuthylazine and S-metolachlor in runoff waters were particularly high in 2007 (2.6% and 0.9% of the amount applied, respectively). Soil half-life of terbuthylazine and S-metolachlor ranged between 12.1 and 8.9 days and 16 and 7 days, respectively. The presence of desethyl-terbuthylazine was related to parent compound degradation. The buffer strip allowed an important reduction of chemical content in water (>90%), in particular during the first runoff events.

Detection, identification, and typing of Listeria species from baled silages fed to dairy cows

Anaerobiosis, critical for successful ensilage, constitutes a challenge in baled silages. The loss of complete anaerobiosis causes aerobic deterioration and silages undergo dry matter and nutrient losses, pathogen growth, and mycotoxin production. Silage may represent an ideal substrate for Listeria monocytogenes, a pathogen of primary concern in several cheeses. The aim of this research was to investigate the occurrence of Listeria in baled silage fed to cows producing milk for a protected designation of origin cheese, and to characterize isolates by repetitive sequence-based PCR. Listeria spp. were detected in 21 silages and L. monocytogenes in 6 out of 80 of the analyzed silages; 67% of positives were found in molded zones. Results of the PCR typing showed genotypic homogeneity: 72.9 and 78.8% similarity between strains of Listeria spp. (n=56) and L. monocytogenes (n=24), respectively. Identical profiles were recovered in molded and nonmolded areas, indicating that contamination may have occurred during production. The application of PCR allowed the unambiguous identification of Listeria isolated from baled silages, and repetitive sequence-based PCR allowed a rapid and effective typing of isolates. Results disclose the potential of the systematic typing of Listeria in primary production, which is needed for the understanding of its transmission pathways.