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The environment temperature and its effect on the temperature of silage is very important for the fermentation and subsequent quality of a silage. Obligate heterofermentative lactic acid bacteria (LAB) inocula, because of their ability to inhibit yeasts, have been developed to prevent the aerobic deterioration of silages. The temperature during silage conservation may also play an important role in the fermentation profile of silages. This study has evaluated the effect of temperature, during the conservation of whole crop corn silage, untreated or treated with different LAB inocula, on the fermentation profile and on the aerobic stability of the silage. Corn was harvested at 42% dry matter and either not treated (control) or treated with Lactobacillus buchneri NCIMB 40788 (LB) at 300,000 cfu/g fresh matter (FM); Lactobacillus hilgardii CNCM I-4785 at 150,000 cfu/g FM (LH150); L. hilgardii CNCM I-4785 at 300,000 cfu/g FM (LH300); or LB+LH at 150,000 cfu/g FM each. In an attempt to experimentally simulate temperature fluctuations in the mass or at the periphery of a silage bunker, corn was conserved in laboratory silos at a constant temperature (20 ± 1°C; MASS) or at lower and variable outdoor temperatures (PERIPH; ranging from 0.5 to 19°C), and the silos were opened after 15, 30, and 100 d of conservation. Lactic acid, acetic acid, and ethanol contents increased in all the silages over the conservation period. The lactic acid content was higher (+10%) in the silages kept at a constant temperature than those conserved at the lower and variable outdoor temperatures. The acetic acid was higher in the treated silages than in the control ones conserved at a constant temperature for 100 d. Moreover, 1,2-propanediol was only detected in the treated silages after at least 30 d at a constant temperature, whereas only traces were detected in the LB+LH treatment for the other temperature conditions. The yeast count decreased during conservation at a slower rate in PERIPH than in MASS and on average reached 2.96 and 4.71 log cfu/g for MASS and PERIPH, respectively, after 100 d of conservation. The highest aerobic stability values were observed for LH300 (191 h) in the MASS silage after 100 d of conservation, whereas the highest aerobic stability was observed in LB+LH (150 h) in the PERIPH silages. After 7 d of air exposure, a pH higher than 4.5 and a higher yeast than 8.0 log cfu/g were detected in all the silages opened after 15 and 30 d of conservation. A pH value close to that of silo opening was detected in the LB, LH150, and LH300 silages conserved under MASS conditions after 100 d, whereas LB+LH was the most effective under PERIPH conditions. The temperature and its fluctuation during conservation of silage in laboratory silos influenced the fermentation, which in turn had an effect on the quality of silage and on the extent of the effect of LAB inocula.

Background Lactic acid bacteria inocula have been developed over the years to improve the aerobic stability of silages. The aims of the study were to evaluate the effect of various conservation periods and the use of Lactobacillus hilgardii inoculum on aerobic stability, fermentative profile and microbial population of corn and sorghum silages. Trials were carried out on two corns and one sorghum crops. The crops were untreated or treated with L. buchneri (LB, application rate 300 000 cfu g-1 FM), L. hilgardii (LH, application rate 300 000 cfu g-1 FM) and a combination (LB + LH, application rate 150 000 cfu g-1 FM each). Silos were opened after 15, 30, 100 and 250 days of conservation, and the silages were analysed for fermentative profile, microbial count and aerobic stability. Results During conservation, the inocula influenced the fermentation profile. The use of LH increased the aerobic stability at 15 and 30 days in one out of three trials, while after 100 and 250 days, the presence of LB alone or with LH led to greater stability. In all the trials, the acetic acid content increased, the yeast count decreased and the aerobic stability increased during the conservation period. Conclusions This study has shown that a long period in complete anaerobiosis reduced yeast count and improved aerobic stability in all silages. The addition of LB was confirmed to be a good option for increasing aerobic stability of silages, whereas the effect of LH alone or in combination with LB on aerobic stability was not consistent between trials. © 2018 Society of Chemical Industry.

The objective of this study was to evaluate the impact of different mixtures of two fresh-cut baby lettuce (Lactuca sativa L. var. crispa cv. Lollo Bionda [LB] and cv. Lollo Rossa [LR]) cultivars on lettuce phytochemical composition during postharvest. Lettuces were grown in a soilless culture system with continuous flotation (FL) in a greenhouse, mixed at harvest and packaged in polypropylene bags and stored at 4 °C for 9 days (d9). Mixes were made of 100, 75, 50, 25 and 0% of LB, respectively. The results showed that the phytochemicals were preserved during storage. In specific, 25LB had the highest pigment content on d1, while 50LB and 25LB had the highest inherent quality on d1.FL led to a reduced microbial contamination, thus, limiting its growth during storage. The results have revealed that high quality and microbiologically safe baby leaf vegetables (BLV), can be obtained by means of FL. The adopting a mix of lettuce cultivars could represent a positive postharvest practice to preserve the phytochemicals of BLV throughout their shelf life.

The growth of Aspergillus flavus and the production of aflatoxins (AF) during the aerobic deterioration of corn silage represent a problem for animal and human health. This experiment was conducted to evaluate whether the presence of A. flavus and AF production originate from the field or additional AF are produced during the fermentation phase or during aerobic deterioration of corn silage. The trial was carried out in northern Italy on corn at a dry matter (DM) level of 34%. The fresh herbage was either not treated (C) or treated with a Lactobacillus buchneri (LB) NCIMB 40788 [(at 3 x 10(5) cfu/g of fresh matter (FM)], Lactobacillus hilgardii (LH) CNCM 1-4785 (at 3 x 10(5) cfu/g of FM), or their combination (LB+LH; at 1.5 x 10(5) cfu/g of FM of each strain) ensiled in 20-L silos and opened after 250 d of ensiling. After silo opening, the aerobic stability was evaluated and samples were taken after 7 and 14 d of air exposure. The pre-ensiled material, the silages at silo opening, and the aerobically exposed silages were analyzed for DM content, fermentative profiles, microbial count, nutritive characteristics, DM losses, and AFB(1), AFB(2), AFG(1), and AFG(2) contents. Furthermore, a subsample of colonies with macromorphological features of Aspergillus section Flavi was selected for AF gene pattern characterization and in vitro AF production. The presence of A. flavus was below the detection limit (

The occurrence of Paenibacillus and Clostridium spores in silage is of great concern for dairy producers because their spores can contaminate milk and damage processed milk and semi-hard cheeses. Spoiled silage is considered to be the main contamination source of the total mixed ration (TMR), feces of dairy cows, and consequently bulk tank milk via the contamination of cow teats by dirt during milking. The presence of an anaerobic and facultative anaerobic sporeformer population in different matrices (soil, corn silage, other feeds, TMR, feces, and milk) and its transmission pathway has been studied on 49 dairy farms by coupling plate count data with 16S-DNA identification. The different matrices have shown a high variability in the anaerobic and facultative anaerobic spore count, with the highest values being found in the aerobically deteriorated areas of corn silages. Clostridium tyrobutyricum, Paenibacillus macerans, and Paenibacillus thermophilus were detected in all the matrices. The TMR spore count was influenced by the amount of spoiled corn silage in the TMR and by the care taken when cleaning the spoiled silage before feed-out. Most of the farms that prevent the presence of visible moldy silage in the silo and carefully clean to remove molded spots were able to maintain their TMR spore counts below 4.0 log spores/g. When a level of 4.5 log spores/g of TMR was exceeded, the feces presented a greater contamination than 3.0 log spores/g. Moreover, the higher the number of spores in the feces was, the higher the number of spores in the milk. Most of the farms that presented a feces contamination greater than 5.0 log spores/g had a higher milk spore contamination than 1,000 spores/L. Careful animal cleaning and good milking practices have been found to be essential to maintain low levels of contamination in bulk tank milk, but it has emerged that only by coupling these practices with a correct silage management and cleaning during TMR preparation can the contamination of milk by spores be kept at a low level. It has been found that aerobically deteriorated silage has a great capacity to contaminate TMR and consequently to increase the risk of milk spore contamination, even when routine milking practices are adopted correctly.

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 m(2) 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 (

Aims 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. Methods and Results Molecular techniques coupled with traditional microbial counting were utilized to identify the predominant fungal species. The cultivable fungal population changed remarkably from harvesting to silo opening. Anaerobiosis and low pH reduced mould count and the presence of Fusarium species both under PE and MB film. However, in the peripheral areas of the silo, where air penetration could not be completely prevented, the fungal population did not decrease. The predominant fungal species after aerobic exposure of silage was Aspergillus fumigatus, without differences between the two plastic films. Conclusions Maintenance of anaerobiosis and a low pH also in the upper layer of the silo reduce the risk of mould growth during corn silage feed-out. Significance and Impact of the Study Even if the new MB plastic film did not completely maintain the anaerobiosis in the upper layer of silage, the overall silage quality was not compromised and was similar to that observed under PE, indicating that the development and use of MB film to cover silage is promising, but needs some improvement.

Lactic acid bacteria inocula have been developed over the years to improve the aerobic stability of silages. The aims of the study were to evaluate the effect of various conservation periods and the use of Lactobacillus hilgardii inoculum on aerobic stability, fermentative profile and microbial population of corn and sorghum silages. Trials were carried out on two corns and one sorghum crops. The crops were untreated or treated with L. buchneri (LB, application rate 300 000 cfu gDuring conservation, the inocula influenced the fermentation profile. The use of LH increased the aerobic stability at 15 and 30 days in one out of three trials, while after 100 and 250 days, the presence of LB alone or with LH led to greater stability. In all the trials, the acetic acid content increased, the yeast count decreased and the aerobic stability increased during the conservation period.This study has shown that a long period in complete anaerobiosis reduced yeast count and improved aerobic stability in all silages. The addition of LB was confirmed to be a good option for increasing aerobic stability of silages, whereas the effect of LH alone or in combination with LB on aerobic stability was not consistent between trials. © 2018 Society of Chemical Industry.

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

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

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.

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.

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.

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 23 °C. A complete suppression of the pathogen was obtained against Monilia sp., stored at 23 °C, and against B. cinerea and P. expansum , stored at 4 °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 1 °C, showed levels of control against B. cinerea and P. expansum similar to those from thiabendazole.