Aerobic Deterioration Of Silage Research Articles

Microbiota succession during aerobic stability of maize silage inoculated with Lentilactobacillus buchneri NCIMB 40788 and Lentilactobacillus hilgardii CNCM-I-4785.

Aerobic deterioration of silage following feeding out is responsible for the deterioration of its quality. Inoculation of silage with lactic acid bacteria is one strategy to limit these effects. A trial was performed using whole‐plant corn ensiled in bag silo, and forage was inoculated with Lentilactobacillus buchneri NCIMB 40788 (Lactobacillus buchneri) and Lentilactobacillus hilgardii CNCM‐I‐4785 (Lactobacillus hilgardii) or not (Control silage). After 159 days of fermentation, the silos were opened and the silage was sampled at 24‐h intervals during a 10‐day aerobic stability assay to measure pH, the fermentation profile, mycotoxins, and microbial and fungal populations. In inoculated silage, lactic acid concentrations and pH remained stable during the aerobic phase and higher microorganism alpha‐diversity was observed. Treated silage was characterized by a high abundance of Saccharomycetes and maintenance of Lactobacillus throughout the aerobic stability assay. The high aerobic stability of the inoculated silage contrasted with the decrease in lactic acid contents and the increase in pH observed in the Control silage, concomitantly with an increase in lactate‐assimilating yeast (Pichia and Issatchenkia), and in Acetobacter and Paenibacillus OTUs. Remarkably, Penicillium and roquefortine C were detected in this silage by day 8 following exposure to air. Our study highlighted the fact that the use of L. buchneri with L. hilgardii modified the consequences of exposure to air by maintaining higher microbial diversity, avoiding the dominance of a few bacteria, and preventing fungi from having a detrimental effect on silage quality.

Aerobic deterioration of corn stalk silage and its effect on methane production and microbial community dynamics in anaerobic digestion

Ensilage is a commonly used method of preserving energy crops for biogas production. However, aerobic deterioration of silage is an inevitable problem. This study investigated the effect of aerobic deterioration on methane production and microbial community dynamics through anaerobic digestion (AD) of maize stalk silage, following 9days air exposure of silage. After air exposure, hydrolytic activity and methanogenic archaea amount in AD were reduced, decreasing the specific methane yield (SMY); whereas lignocellulose decomposition during exposure improved the degradability of silage in AD and enhanced SMY, partially compensating the dry matter (DM) loss. 29.3% of the DM and 40.7% of methane yield were lost following 0–9days exposure. Metagenomic analysis showed a shift from Clostridia to Bacteroidia and Anaerolineae in AD after silage deterioration; Methanosaetaceae was the dominant methanogenic archaea.

Characterization of culturable yeast species associating with whole crop corn and total mixed ration silage.

ObjectiveThis study investigated the association of yeast species with improved aerobic stability of total mixed ration (TMR) silages with prolonged ensiling, and clarified the characteristics of yeast species and their role during aerobic deterioration.MethodsWhole crop corn (WCC) silages and TMR silages formulated with WCC were ensiled for 7, 14, 28, and 56 d and used for an aerobic stability test. Predominant yeast species were isolated from different periods and identified by sequencing analyses of the 26S rRNA gene D1/D2 domain. Characteristics (assimilation and tolerance) of the yeast species and their role during aerobic deterioration were investigated.ResultsIn addition to species of Candida glabrata and Pichia kudriavzevii (P. kudriavzevii) previously isolated in WCC and TMR, Pichia manshurica (P. manshurica), Candida ethanolica (C. ethanolica), and Zygosaccharomyces bailii (Z. bailii) isolated at great frequency during deterioration, were capable of assimilating lactic or acetic acid and tolerant to acetic acid and might function more in deteriorating TMR silages at early fermentation (7 d and 14 d). With ensiling prolonged to 28 d, silages became more (p<0.01) stable when exposed to air, coinciding with the inhibition of yeast to below the detection limit. Species of P. manshurica that were predominant in deteriorating WCC silages were not detectable in TMR silages. In addition, the predominant yeast species of Z. bailii in deteriorating TMR silages at later fermentation (28 d and 56 d) were not observed in both WCC and WCC silages.ConclusionThe inhibition of yeasts, particularly P. kudriavzevii, probably account for the improved aerobic stability of TMR silages at later fermentation. Fewer species seemed to be involved in aerobic deterioration of silages at later fermentation and Z. bailii was most likely to initiate the aerobic deterioration of TMR silages at later fermentation. The use of WCC in TMR might not influence the predominant yeast species during aerobic deterioration of TMR silages.

Aerobic stability of sugar‐cane silage inoculated with tropical strains of lactic acid bacteria

AbstractAerobic stability is an important feature in the evaluation of silages. The aims were to investigate the chemical and microbiological changes that occur in sugar‐cane (Saccharum spp.) silage after aerobic exposure, to identify the major species of yeasts associated with the aerobic deterioration process and to select lactic acid bacteria (LAB) strains that can improve the aerobic stability of this silage. Fourteen wild LAB strains belonging to Lactobacillus plantarum, L. brevis and L. hilgardii were evaluated using experimental silos. Silage samples were collected at 0, 96 and 216 h after aerobic exposure to determinate the DM, WSC, pH, products of fermentation, to evaluate the silage temperatures and to identify yeast species associated with the aerobic deterioration of silage. The strains tested were able to modify the fermentative and chemical parameters and the diversity of yeasts species of silage after aerobic exposure. There was no association between the facultative or obligatory heterofermentative fermentation patterns and the increased aerobic stability of silage. Aerobic stability of sugar‐cane silages was associated with high acetic acid and 1,2‐propanediol concentrations. L. hilgardii UFLA SIL51 and UFLA SIL52 strains promoted an increase in aerobic stability of silage.

Determination of aerobic deterioration of corn stalk silage caused by aerobic bacteria

Aerobic deterioration of silage is an important problem in silage production because it affects not only the silage quality but also the animal health. The objective of the current study was to investigate the aerobic stability of corn stalk silage and the role of microbes resulting in aerobic deterioration. Corn stalks were ensiled in evacuated plastic bags for 65 days at room temperature. After the bags were opened, the silages were subjected to treatment with chloramphenicol (CL) or without CL (WCL). The aerobic bacteria and yeast compositions of silages at opening and after exposure to air for 7 days were investigated by analyzing the sequence divergence in the 16S ribosomal DNA gene and in the D1/D2 region of the 26S ribosomal DNA gene, respectively. The results showed that corn stalk silages were well preserved, indicated by the low pH value and high lactic acid content. At the opening of bags, the aerobic bacteria in corn stalk silage consisted of Myroides odoratimimus, Acinetobacter soli, Stenotrophomonas maltophilia, Serratia nematodiphila, Bacillus pumilus and Lysinibacillus fusiformis, but contained undetectable amounts of yeasts (<2.0010logcfu/g FM). After exposure to air for 7 days, WCL silage deteriorated, while CL silage did not, because aerobic bacteria were inhibited by CL. Further study showed that L. fusiformis were predominant in WCL silage (0.625 of aerobic bacteria) and metabolized lactic acid. Therefore, in the present study, L. fusiformis resulted in aerobic deterioration of corn stalk silage.

Silage and the safety and quality of dairy foods: a review

Silage contains a number of potential hazards to the safety and quality of milk and dairy products. This paper reviews the present knowledge about silage as a source of (1) spores of anaerobic spore-formers (Clostridium species) and aerobic spore-formers (mainly Bacillus and Paenibacillus species), (2) the zoonotic pathogenic bacteria Listeria monocytogenes and Escherichia coli, and (3) mycotoxins. A distinction is made between field-derived mycotoxins, i.e. mycotoxins that are formed during growth of crops in the field, and ensilage-derived mycotoxins, i.e. mycotoxins that are formed after ensiling. The routes of transmission of these hazards from feed to milk, the effect of pasteurization of milk, and reduction strategies are discussed. Aerobic deterioration of silages is a major factor influencing levels of spores of both aerobic and anaerobic spore-formers, L. monocytogenes, and certain mycotoxins.

Dry matter and nutritional losses during aerobic deterioration of corn and sorghum silages as influenced by different lactic acid bacteria inocula

The economic damage that results from aerobic deterioration of silage is a significant problem for farm profitability and feed quality. This paper quantifies the dry matter (DM) and nutritional losses that occur during the exposure of corn and sorghum silages to air over 14 d and assesses the possibility of enhancing the aerobic stability of silages through inoculation with lactic acid bacteria (LAB). The trial was carried out in Northern Italy on corn (50% milk line) and grain sorghum (early dough stage) silages. The crops were ensiled in 30-L jars, without a LAB inoculant (C), with a Lactobacillus plantarum inoculum (LP), and with a Lactobacillus buchneri inoculum (LB; theoretical rate of 1 × 106 cfu/g of fresh forage). The pre-ensiled material, the silage at silo opening, and the aerobically exposed silage were analyzed for DM content, fermentative profiles, yeast and mold count, starch, crude protein, ash, fiber components, 24-h and 48-h DM digestibility and neutral detergent fiber (NDF) degradability. The yield and nutrient analysis data of the corn and sorghum silages were used as input for Milk2006 to estimate the total digestible nutrients, net energy of lactation, and milk production per Mg of DM. The DM fermentation and respiration losses were also calculated. The inocula influenced the in vitro NDF digestibility at 24h, the net energy for lactation (NEL), and the predicted milk yield per megagram of DM, whereas the length of time of air exposure influenced DM digestibility at 24 and 48h, the NEL, and the predicted milk yield per megagram of DM in the corn silages. The inocula only influenced the milk yield per megagram of DM and the air exposure affected the DM digestibility at 24h, the NEL, and the milk yield per megagram of DM in the sorghum silages. The milk yield, after 14 d of air exposure, decreased to 1,442, 1,418, and 1,277 kg/Mg of DM for C, LB, and LP corn silages, respectively, compared with an average value of 1,568kg of silage at opening. In the sorghum silages, the milk yield, after 14 d of air exposure, decreased to 1,226, 1,278, and 1,250 kg/Mg of DM for C, LB, and LP, respectively. When the estimated milk yield per megagram of harvested DM of corn and sorghum silage were related to mold count, it was shown that the loss of potential milk production occurred when the mold count exceeded 4 log cfu/g of silage, and it was almost halved when the mold count reached values greater than 8 log cfu/g of silage. Inoculation with L. buchneri, at a rate of 1 × 106 cfu/g of fresh forage, enhanced the stability of the silage after exposure to air, and, consequently, contributed to maintaining the nutritional value of the harvested forage over time, for air exposure up to 7 d.

Anaerobic lactic acid degradation during ensilage of whole crop maize inoculated with lactobacillus buchneri inhibits yeast growth and improves aerobic stability

Aerobic deterioration of silages is initiated by (facultative) aerobic micro-organisms, usually yeasts, that oxidize the preserving organic acids. In this study, a Lactobacillus buchneri strain isolated from maize silage was evaluated for its potential as a bacterial inoculant that enhances aerobic stability of silages. In four experiments, chopped whole crop maize (30-43% dry matter (DM)) was inoculated with Lact. buchneri and ensiled in laboratory silos. Uninoculated silages served as controls. Analysis of silages treated with Lact. buchneri at levels of 103-106 cfu g-1 after about 3 months of anaerobic storage showedthat acetic acid and 1-propanol contents increased with inoculum levels above 104 cfu g-1,whereas lactic acid decreased. Propionic acid, silage pH and DM loss increased withinoculum levels above 105 cfu g-1. Time course experiments with maize inoculated with Lact. buchneri at 4 x 104-2 x 105 cfu g-1 showed that up to 7-14 d after ensiling, Lact. buchneri had no effect on silage characteristics. Thereafter, the lactic acid content of the inoculated silages declined and, simultaneously, acetic acid and, to a lesser extent, propionic acid and 1-propanol, accumulated. Inoculation reduced survival of yeasts during the anaerobic storage phase and inhibited yeast growth when the silage was exposed to O2, resulting in a substantial improvement in aerobic stability. The results indicate that the use of Lact. buchneri as a silage inoculant can enhance aerobic stability by inhibition of yeasts. The ability of the organism to ferment lactic acid to acetic acid appears to be an important underlying principle of this effect.

Effect of applying lactic acid bacteria isolated from forage crops on fermentation characteristics and aerobic deterioration of silage.

Two selected strains, Lactobacillus casei FG 1 and Lactobacillus plantarum FG 10 that were isolated from forage crops were used as additives at 1.0 x 10(5) cfu/g of fresh matter to alfalfa, Italian ryegrass, and sorghum, and their effect on fermentation characteristics and aerobic deterioration of silage was studied. The three silages treated with strains FG 1 or FG 10 were well preserved; had significantly lower pH values, butyric acid, propionic acid, and ammonia N concentrations, gas production, and dry matter losses; and had significantly higher contents of residual water-soluble carbohydrates and lactic acid than did the respective control silages. Yeast counts were high in all treated silages and increased rapidly during aerobic exposure. As a result, treated silages spoiled faster upon aerobic exposure than did the respective control silages. Most yeasts isolated from deteriorated silages showed high tolerance to lactic acid but low tolerance to butyric acid, and they were able to grow at low pH conditions and assimilate lactic acid. The results confirmed that L. casei and L. plantarum improved fermentation quality but did not inhibit the growth of silage yeast or aerobic deterioration of the silage.

Construction of Kluyveromyces lactis killer strains defective in growth on lactic acid as a silage additive

Killer strain of Kluyveromyces lactis was constructed to prevent the aerobic deterioration of silage by disruption of KlPCK1 gene encoding phosphoenolpyruvate carboxykinase (PEPCK). The disruptants are defective in their ability of growth on lactic acid as a sole carbon source. The PEPCK activity was also deficient in the disruptants. The growth rate on lactose medium and the killing activity were equal to those of the parental strain. © Rapid Science Ltd. 1998

Selection of Killer Yeasts (Kluyveromyces lactis) to Prevent Aerobic Deterioration in Silage Making

Killer spectra were investigated to select killer yeast strains to inhibit the growth of wild yeasts that cause aerobic deterioration of silage. Kluyveromyces lactis IFO 1267 was characterized by its rapid killing activity and wide spectrum against the target yeasts. Crude killer protein produced by K. lactis IFO 1267 or inoculation of the cell itself inhibited the growth of the target strain, Saccharomyces cerevisiae IFO 0304 (in liquid or in solid culture), particularly when lactose was used as carbon source instead of glucose.

Microbial dynamics during aerobic deterioration of silages

AbstractFactors affecting microbial dynamics during aerobic deterioration of silages were evaluated in laboratory, pilot‐scale and full‐scale experiments. Yeasts usually caused the initial deterioration but in some samples with fairly good storage stability (&gt;10 days) the activity was associated with the growth of moulds and bacteria. At a later stage, and especially at higher temperatures (&gt;40°C), the activity was caused by bacilli. The increase in pH as a result of lactate metabolism permitted the growth of Enterobacteria. Evaluation of storage stability in samples from different levels in a bunker silo indicated low stability in surface samples and fairly good stability in samples from 160cm depth. The surface samples contained higher numbers (100–1000 times) of yeasts but the pH levels were similar.The establishment of yeasts during storage is probably due to diffusion of oxygen by leakage through the plastic cover. Compared with a plastic cover (0.1mm polythene), a rubber seal improved storage stability in laboratory silage (pH4.0) inoculated with lactic acid bacteria and infected with Candida albicans. An increase in acid content and a reduction in pH (3.7) by means of an efficient inoculant did not seem to increase the storage potential under the conditions described above. The content of adenosine triphosphate has been used as a measure of microbial activity during aerobic deterioration of silages. The main activity was concentrated at the surface (0–5cm). At 20°C, the activity was about ten times higher at the surface than at a depth of 30cm; above 30°C it was about 100 times higher.

Investigations into the role of specific micro-organisms in the aerobic deterioration of maize silage

SummaryTwo experiments were undertaken with maize silage, which had been sterilized by subjecting it to a 6 Mrad dose of γ radiation, inoculated with micro-organisms implicated in the aerobic deterioration of silage and subsequently exposed to air. The inocula employed comprised mixed strains of yeasts, bacilli and lactic acid bacteria and combinations of these. The changes which accompanied aerobic deterioration were observed. The major losses of fermentation acids, ethanol, ammonia and dry matter, and increases in total nitrogen and temperature were associated with inocula in which yeasts were represented. Similar changes of a lower magnitude were also observed in silage treated with inocula containing bacteria only, although these were not always accompanied by the growth of these organisms or loss of dry matter. However, when employed in mixed inocula with yeasts, the bacteria proliferated and tended in some instances to increase losses further, suggesting an association between the groups of micro-organisms in the aerobic deterioration process. Losses were also sustained in sterile silage which received no inoculum and the possible reasons for this are discussed.

Gamma radiation in the preparation of maize silage for studies on the aerobic deterioration process

SUMMARYMaize silage was exposed to doses of y radiation varying from 0 to 6 Mrad inclusive and the effects of this treatment on the microflora and possible changes it might impose on the chemical composition were monitored. The highest dose was required to reduce the native population of micro-organisms to an insignificant level and only minor losses of lactic acid and ethanol were sustained. In addition, the irradiated silage was inoculated with representatives of groups of micro-organisms implicated in the aerobic deterioration of silage and the subsequent changes upon exposure to air observed. The inoculum proliferated and induced losses of fermentation acids, water-soluble carbohydrates, dry matter and, to a minor degree, neutral detergent fibre, and resulted in an increase in temperature and total nitrogen, the factors most likely to change during aerobic deterioration. Only the 6 Mrad dose yielded material suitable for investigations into the identification of those organisms primarily responsible for aerobic deterioration.

Aerobic Deterioration of Wheat, Lucerne and Maize Silages Prepared with Lactobacillus acidophilus and a Candida spp.

Aerobic deterioration of lucerne, maize and wheat silages was characterized by rapid increases in yeast and mould flora which oxidized lactic and volatile acids resulting in increased temperature and pH. While populations of yeasts and moulds were similar, temperature increases were slightly greater for silages inoculated with Lactobacillus acidophilus and Candida spp. After 48 h the pH of the inoculated silages was higher in general and concentrations of acids were lower than controls. Bacterial growth was slight although continued lactic acid production was probable. In contrast to lucerne and maize silages, the pH of wheat silage remained stable during this period because of high butyric levels, but temperature and yeast populations increased. After 48 h the pH rose above 5 in maize and lucerne, and bacterial growth and metabolic activity resumed resulting in volatile and non‐volatile acid production from carbohydrate fermentation and deamination of amino acids. During this phase of aerobic deterioration yeast growth slowed or stopped, but temperatures remained high and pH continued to climb probably because of production of ammonia. The changes in gross composition of the silages did not follow any particular pattern. Losses in dry matter were small (2.5–4.0%) and changes in individual components probably reflect this loss rather than substantial changes. Protein availability in the lucerne silages undoubtedly decreased, as protein losses were high. It is concluded that the aerobic deterioration of silage is enhanced by the addition of L. acidophilus and Candida spp. at ensiling.

サイレージ変敗防止剤の添加効果と微生物相-材料水分ならびに薬剤抵抗性菌との関係について

サイレージ変敗防止剤の添加効果と微生物相-材料水分ならびに薬剤抵抗性菌との関係について

The effect of some additives on aerobic deterioration of silages

AbstractTwo experiments are described. In the first, formic acid was applied alone and in combination with isovaleric, caproic or lauric acid to wilted herbage prior to ensiling. The effects of these treatments on the deterioration of silages on exposure to air over a 7 day period were measured. Temperature increase, chemical and microbiological changes were determined. Silages treated with formic acid alone deteriorated, while the untreated control silages were relatively stable. The formic acid treated silages containing either isovaleric or caproic acid remained stable over the 7 day aerobic period. In the second experiment, in addition to the previous treatments, formaldehyde was also included in place of formic acid. In this case, only the untreated control silages deteriorated markedly over the 7 day period.

サイレージから分離した酵母の各種培地における増殖とサイレージの好気的変敗との関係

サイレージから分離した酵母の各種培地における増殖とサイレージの好気的変敗との関係