枇杷汁乳酸发酵优良菌株筛选及其发酵特性研究

以枇杷汁为原料,以乳酸发酵风味为主要评判指标,采用纯种控温发酵技术,筛选枇杷果汁乳酸发酵的优良乳酸菌,并研究其发酵特性。实验结果表明,枇杷果汁乳酸发酵的最适菌株为植物乳杆菌R23,其次为干酪乳杆菌R35,且2者发酵风格各具特色。发酵温度低于30℃时,菌细胞生长有较长的稳定期。枇杷果汁发酵过程中,总酸先降后升,酸代谢增减速率与发酵温度呈正相关。植物乳杆菌R23最适发酵温度25～30℃,发酵时间2～3 d。原果酸含量对发酵风味影响不显著。     

二步发酵法在西番莲果皮酵素加工中的应用

以西番莲果皮为对象,研究酿酒酵母菌、乳酸菌二步发酵工艺,制备乳酸菌和酿酒酵母菌活菌数高、多酚保留好、风味醇和的果皮浆。通过单因素和正交优化实验,以酿酒酵母菌和乳酸菌活菌数、总酚、乙醇生成量为指标,分别优选了单一乳酸菌和酿酒酵母菌发酵的工艺参数,在此基础上,通过测定总酸含量、活菌数、总酚含量等指标的变化,确定二步法发酵西番莲果皮浆的工艺参数为：发酵总时间40 h,酿酒酵母菌初始接种量1.5%,在29 ℃下发酵16 h后,再接入2.0%复合乳杆菌（干酪乳杆菌∶植物乳杆菌=1∶1）,在37 ℃下继续发酵24 h至终点。制备的西番莲果皮发酵液中,复合乳杆菌活菌数为8.93 lg CFU/mL,酿酒酵母菌活菌数为7.78 lg CFU/mL,总酚含量为24.00 μg/mL,总酸含量为4.40 mg/mL。二步发酵法发挥了酿酒酵母菌产香、产乙醇抑制杂菌,以及乳酸菌提高酸度、保留多酚物质的优点。     

Microbial and Biochemical Changes Occurring During Production of masvusvu and mangisi,traditional Zimbabwean beverages

Biochemical and microbiological changes that occur during production of masvusvu and mangisi, both traditional Zimbabwean beverages made from malted finger millet flour were investigated in this study. During cooking to produce masvusvu, amylase was active within the first 20 minutes but its activity decreased drastically at temperatures of above 60 °C. Free reducing sugars increased from 3.8 to 308.1 mg/ml. During fermentation to produce mangisis, total aerobic mesophilic bacteria, lactic acid bacteria and yeast and molds increased with fermentation time. Total titratable acidity increased from 0.13 to 0.67 percent, lactic acid from 0.51 t 4.10 g/l and pH decreased from 6.10 to 3.98.     

香蕉果酒接种发酵和自然发酵过程中乳酸菌菌落变化及有机酸代谢特征

香蕉营养丰富、活性物质含量高，可作为深加工产品的优质原料。香蕉果酒是香蕉精深加工产品中的重要一类。由于香蕉果浆碳水化合物含量较高、酸度低、糖分高，其酿造过程中适宜各种微生物的生长繁殖，特别是乳酸菌数量的变化与酸类物质的产生密切相关。乳酸菌群落的变化容易增加发酵过程的风险，造成香蕉酒挥发酸含量超标、品质低下等不良结果。因此明确香蕉果酒发酵过程中自然存在的乳酸菌的生长繁殖规律及其对有机酸代谢的影响，可为精准调控发酵过程、提高产品质量提供理论依据。试验设置商业酵母Lalvin K1^TM接种发酵过程（K1）与香蕉果汁自然发酵过程（SF）2组对比，采用实时定量PCR方法和高效液相色谱法对香蕉汁接种发酵（K1）与自然发酵过程（SF）中酵母菌、乳酸菌的生长规律以及有机酸的含量变化进行监测。结果表明：2种发酵处理下，酵母菌和乳酸菌的生长规律有差异；K1酵母菌主导的发酵过程中，乳酸菌的生长受到显著抑制，最大生长量仅为4.0×10⁶ CFU/mL，而酵母菌生物量为1.2×10⁷ CFU/mL；在自然发酵过程中，乳酸菌与酵母菌的生长周期完全错开，但对乳酸菌的生物量无显著影响，乳酸菌与酵母菌的生长量分别达到1.0×10⁷ CFU/mL和1.6×10⁷ CFU/mL。2种发酵过程均表明，酵母菌和乳酸菌的生长存在竞争与抑制关系；有机酸代谢主要与乳酸菌菌落数量相关，不同有机酸的总体变化趋势一致；在发酵过程中，乳酸含量逐渐增加，乙酸含量呈先增后降的趋势，二者在自然发酵样品中的累积量均高于接种发酵；柠檬酸、苹果酸、酒石酸含量整体呈下降趋势。     

Influence of harvest date of primary growth on microbial flora of grass herbages and haylage, and on fermentation and aerobic stability of haylage conserved in laboratory silos

The influence of harvest date of the primary growth of grass herbages on the microbial flora of herbage pre-conservation and haylage post-conservation was studied along with fermentation variables and aerobic stability of haylage. The primary growths of two grass swards, one intensely (Int) and one extensively (Ext) managed, were cut at three different harvest dates (May, June, August) and ensiled in laboratory silos for 120 d. Later harvest dates resulted in increased counts of yeast, mould and enterobacteria in the pre-conserved herbage ( P  <   0·001). Counts of lactic acid bacteria (LAB) ( P  <   0·001) and number of mould species ( P  <   0·001) were highest in the pre-conserved herbage harvested in August. Later harvest dates resulted in higher yeast ( P  <   0·001) and LAB ( P  <   0·001) counts in the haylage while counts of enterobacteria decreased ( P  <   0·001). Clostridial spore counts were unaffected by harvest date both in herbage and haylage. The haylage harvested in August had the lowest pH and the lowest concentration of ethanol but the concentration of lactic acid was in general low. Aerobic stability was longer ( P  <   0·01) for haylage from herbage harvested in August compared with haylage from herbage harvested in May and June. Sward type had less influence than harvest date on microbial variables in herbage and haylage and on fermentation variables of haylage, and did not influence the aerobic stability of haylage.     

Fermentation and nutritive value of Zea mays and lucerne forage ensiled with added rolled barley

The effect of adding rolled barley on the ensiling characteristics and nutritive value of lucerne and corn ( Zea mays ) forage was studied using 19-1 plastic mini-silos. Other factors evaluated with lucerne were the effect of wilting and added lactic acid bacteria. Rolled barley was added at rates of 0, 50, 100 and 150gkg⁻¹ (wet weight) to lucerne or corn forage. Lucerne was ensiled at 170–330 g kg⁻¹ dry matter (DM) and lactic acid bacteria were applied at 10⁵g⁻¹ wet forage. Addition of rolled barley consistently improved the fermentation of lucerne by lowering the pH and decreasing the concentration of acetate and ammonia nitrogen. Wilting of lucerne had variable effects on fermentation in two experiments. The addition of lactic acid bacteria improved fermentation by decreasing the concentrations of acetate and ammonia nitrogen, improved the nutritive value by increasing the in vitro dry matter digestibility (IVDMD), and was associated with decreased amounts of lactic acid. Corn was ensiled at 250 g kg⁻¹ DM, and added rolled barley had little effect on the fermentation of corn silage. The digestibility of all silages determined in vitro was improved by the addition of rolled barley.     

RESEARCH NOTE

Data from twenty-two experiments conducted at tour ADAS Research Centres during 1980–92 were used to compare untreated silages with silages treated with formic acid, with or without added formalin, commercial inoculants or molasses. The sillages were made from herbage whose dry-matter (DM) and water-soluble carbohydrate (WSC) contents were 277 (s.e. 0.46) g DM kg⁻¹ and 36 (s.e. 8.1) g kg⁻¹ respectively. Inoculant use significantly decreased silage pH and ammonia-N, significantly increased lactic acid and total acid content, and decreased butyric acid and total short-chain fatty acids. Formic acid use significantly increased silage lactic acid and total acid content, and decreased butyric acid content, whereas formic acid+formalin significantly decreased silage ammonia-N level. Molasses had little effect upon silage fermentation. Improvements in silage fermentation, however, produced little benefit in terms of either silage DM intake or liveweight gain when the silages were offered to growing lambs.
It is suggested from the results that inoculant- and formic acid-based additives can be used to improve the fermentation of big-bale silages.     

Effect of chitosan on the preservation quality of sugarcane silage

We aimed to evaluate the effects of chitosan and microbial inoculant addition to sugarcane silage fermentation, gas and effluent losses, chemical composition, in situ dry matter (DM), neutral detergent fibre (NDF) degradation and aerobic stability. A completely randomized design with four treatments (n = 40) was performed. It was arranged in a 2 × 2 factorial scheme with chitosan [0 and 6 g/kg of sugarcane DM—1.66 g/kg of natural matter (NM)] and microbial inoculant (0 and 8 mg/kg on NM). Each g of inoculant contained 3.9 × 10¹⁰ UFC/g of Pediococcus acidilactici and 3.75 × 10¹⁰ UFC/g of Propionibacterium acidicipropionici. The addition of microbial inoculant increased lactic acid concentration in silos treated with chitosan. Furthermore, chitosan increased pH and tended to increase acetic acid of silage. In contrast, the inoculant decreased pH and acetic acid, besides increasing ethanol concentration. As chitosan addition increased DM recovery, inoculant addition decreased it. Chitosan decreased NDF and acid detergent fibre (ADF) level and increased DM degradation, while inoculant decreased DM content, DM and NDF degradation. In addition, chitosan improved the aerobic stability only in non‐inoculated silos. Thus, chitosan has a positive effect on silage fermentation, reducing fermentative losses, and improving silage chemical composition and degradation. Conversely, the addition of microbial inoculant negatively affected silage DM recovery, chemical composition, and its association with chitosan decreased the aerobic stability when compared to the exclusive use of chitosan.     

Integrated utilisation of green biomass in the green biorefinery

Grass, clover and alfalfa are used for production of fodder pellets. In the green crop drying factories in the northern countries the drying is combined with pressing of the green crops resulting in production of large volumes of plant juices, green and brown juice. In order to get rid of these enormous amounts of plant juice, representing an environmental threat, it is our aim to convert the simple drying industry to a whole crop utilisation factory, a green biorefinery, where the plant juice can be used as raw material in a fermentation process. As the plant juice is very unstable, it has been necessary to develop a method for both conservation and utilisation of the juice. The plant juice is converted to a nice universal fermentation medium by lactic acid bacteria. A number of lactic acid producing strains have been tested. Among the most productive strains, Lactobacillus paracasei ssp. paracasei P4155 is found to be the most effective in utilising fructans and citric acid. Most carbohydrates and organic acids in the juice can be converted to lactic acid by two homofermentative lactobacilli. The highest lactic acid yield and productivity is reached when using not-heat sterilised brown or green juice as the fermentation medium. The amount of free amino acids in the juice is increased and the content of vitamins and other important growth factors kept unspoiled by the lactic acid fermentation. The fermented juice can be stored as it is or evaporated to a concentrated form and used in fermentation industries for production of amino acids, organic acids, enzymes etc.     

In vitro rumen methane output of grasses and grass silages differing in fermentation characteristics using the gas‐production technique (GPT)

The chemical composition of silage consumed by cattle can influence the subsequent rumen microbial fermentation patterns and methane (CH₄) emissions. The objectives of this study were to (i) evaluate the effect of ensilage on the in vitro rumen methane output of perennial ryegrass and (ii) relate the silage fermentation characteristics of grass silages with in vitro rumen methanogenesis. Three pre‐harvest herbage‐conditioning treatments and seven silage‐additive treatments were used in a laboratory‐scale silo experiment to produce a diversity of silage fermentation characteristics. Ensilage reduced (P < 0·01) the in vitro rumen CH₄ output (mL CH₄ g^?1 dry matter (DM) disappeared). This reflected differences in the direction of rumen fermentation (lower acetic (P < 0·05) and higher propionic (P < 0·001) acid proportions in volatile fatty acids) rather than major changes in the extent of in vitro rumen fermentation (i.e. mmol VFA g^?1 DM). The magnitude of the decrease in CH₄ output (mL g^?1 DM incubated) owing to ensilage increased as the extent of silage fermentation, and in particular the lactic acid concentration, increased. In contrast, among silages with relatively similar extents of silage fermentation (i.e. total fermentation products), an increase in the proportion of lactic acid in silage fermentation products led to a more extensive in vitro rumen fermentation and thus to a greater CH₄ output (mL g^?1 DM).     

Separating the effects of forage source and field microbiota on silage fermentation quality and aerobic stability

This study attempted to separate the effects of forage source and field microbiota on silage fermentation quality and aerobic stability. Single samples of grass, red clover and maize were used. Field microbiota was obtained by centrifugation of microbial suspensions of the three samples. The intact forages were dried and sterilized by heating at 60°C for 3 h + 103°C for 15 h, inoculated in a 3 (forage) × 3 (inoculum) design and reconstituted to a dry‐matter level of 400 g kg^?1 before ensiling. After ensiling for 71 d, subsamples were subjected to an 8‐d aerobic stability test, which included temperature and pH measurements. Bacterial community analysis was performed on samples before and after ensiling by 16S rRNA gene amplicon sequencing. Forage source had a marked effect on the levels of lactic acid, acetic acid, ammonia‐N and 2,3‐butanediol, but microbiota source only affected the acetic acid concentration. The forage and microbiota as well as their interactions affected silage stability variables. The maize microbiota improved silage stability, whereas silages made from the maize forage had the poorest stability. Bacterial community analysis revealed higher abundance of lactic acid bacteria on the maize forage, with Lactococcus and Leuconostoc being the dominant genera. These preliminary results suggested that fermentation quality is mainly affected by forage source, whereas the aerobic stability is affected by both forage and field microbiota.     

Fermentation by selected sourdough lactic acid bacteria to decrease coeliac intolerance to rye flour

A pool of selected lactic acid bacteria was used to ferment suspensions of rye flour. Two-dimensional electrophoresis showed that 109 of the 129 ethanol-soluble rye polypeptides were hydrolysed almost totally by lactic acid bacteria. Matrix-assisted laser desorption ionization—time of flight mass spectrometry and reversed-phase high performance liquid chromatography analysis confirmed the hydrolysis of prolamins. After 48 h fermentation, no prolamin polypeptides were recognized by R5-Western analysis. HPLC analysis of glutelin polymers showed a very low bacterial proteolysis but a pH dependent hydrolysis probably due to activation of rye enzymes. Prolamins were extracted from rye flour and used to produce a peptic–tryptic (PT)-digest for in vitro tests with K 562 (S) sub-clone and Caco-2/TC7 cells of human origin. The PT-digest was also treated with lactic acid bacteria before assay. The Minimal Agglutinating Capacity increased ca. 8-times when K 562 (S) sub-clone cells were exposed to rye PT-digest treated with lactic acid bacteria. Hydrolysis of rye PT-digest by lactic acid bacteria decreased the toxicity of PT-digest itself towards Caco-2/TC7 cells as estimated by cell viability, caspase-3 activity and release of nitric oxide. Rye prolamins and glutelins were extracted from doughs and subjected to PT digestion. Compared to PT-digests from chemically acidified dough, coeliac jejunal biopsies exposed to the PT-digest from the dough fermented by lactic acid bacteria did not show an increase of the infiltration of CD3⁺ intraepithelial lymphocytes. The same was found for epithelial cell Fas expression. Long-time fermentation of dough by selected lactic acid bacteria could be considered as a potential tool to decrease the risk of rye contamination of gluten-free products for coeliac patients.     

Effects of Maize Type and Fermentation Conditions on the Quality of Beninese Traditional Ogi,a Fermented Maize Slurry

Four Beninese cultivars (two local ecotypes and two new cultivars) were tested for their ability to produce ogi. The two local ecotypes, having friable and floury to medium vitreous kernels, gave a higher yield of ogi (10% more) than the improved cultivars that had coherent and vitreous kernels. They also gave slightly more viscous ogi porridges. Whatever the storage conditions (20 to 35°C, 3 to 15 d), a hetero-lactic fermentation generated two characteristic products: lactic acid and ethanol. However, the fermentation process was more intense and more rapid at 35°C giving a peak in acidity at the end of the first week, then a decrease during the second week. In parallel, ori porridge viscosity increased during the second week of fermentation.     

Modification of the rheological behavior of amaranth (Amaranthus hypochondriacus) dough

For people with celiac disease, a lifelong abdication of gluten including-products is necessary to live a life without celiac affected reactions. The production of high-quality bread from gluten free flour is not simple in comparison to gluten including flours such as those derived from wheat (Triticum spp.). The gas binding and crumb structure forming capacity are very low in gluten free batters. They can efficiently be analyzed through the rheological properties of the dough used. The use of acidification in amaranth (Amaranthus hypochondriacus) dough preparation is a possible means of changing the rheological behavior of amaranth in the desired direction. Methods include the use of lactic acid directly, or the fermentation via lactic acid bacteria. Adding up to 20 mL lactic acid/kg flour in amaranth dough preparation led, during oscillation tests, to an increase of the complex shear modulus up to 30% in the range of 0.1 up to 10 Hz. The use of sourdough fermentation decreased the complex shear modulus in the same test up to nearly 60%. In creep recovery tests, the elastic part of amaranth dough decreased from 65.4% without any treatment down to 63.9% by the addition of up to 20 mL lactic acid/kg flour. Sourdough fermentation by Lactobacillus plantarum was able to decrease it to 54%. The acidification showed a significant positive influence on the rheological parameters of amaranth dough only at the higher stress level. In contrast, sourdough fermentation was able to produce doughs with viscosity and elasticity similar to that found in pure wheat flours.     

Effects of inoculants and wilting on silage fermentation and nutritive characteristics of red clover–grass mixtures

Various management practices (e.g. wilting, application of silage additives or adding a grass component) can be used to improve silage fermentation of pure red clover (Trifolium pratense L.). Therefore, the aim of this laboratory ensiling study was to investigate the effects of varying proportions of red clover and perennial ryegrass (100/0, 66/33, 33/66, 0/100) on silage quality during two consecutive years. In addition, two wilting levels [target dry matter (DM): 300 vs. 400 g kg^?1] in combination with lactic acid bacteria (LAB) additives were tested. Herbage was ensiled, either untreated or inoculated with homofermentative LAB (low wilted) or homo‐ and heterofermentative LAB (high wilted). In most cases, lactic and acetic acid decreased as the proportions of ryegrass were increased. Data concerning ammonia‐N concentrations showed considerable differences between cuts and years. Silages treated with homofermentative LAB generally had high lactic acid and low final pH, whereas acetic acid and 1,2‐propanediol tended to be higher when homo‐ and heterofermentative LAB were applied. Inoculants had a positive effect on DM losses and ammonia‐N in only a few silages. Wilting decreased DM losses and fermentation acids at most cuts, irrespective of the grass/clover ratio in the herbage mixture. There was a strong year effect on the organic matter digestibility (DOM) of the silages. In conclusion, the optimal strategy for successful silage fermentation of red clover is the ensiling in mixtures with ryegrass. Furthermore, herbage should be wilted to a DM content of about 300–350 g kg^?1. The application of LAB inoculants did not alter the DOM but did improve silage fermentation.     

Silage fermentation characteristics of three common grassland species in response to advancing stage of maturity and additive application

This study investigated the effects of advancing stage of maturity and additive treatment on the fermentation characteristics of three common grassland species. Perennial ryegrass (PRG; Lolium perenne L., var. Gandalf), timothy (Phleum pratense L., var. Erecta) and red clover (Trifolium pratense L., var. Merviot) were harvested at three dates in the primary growth (11 May, 8 June and 6 July; Harvests 1–3) and ensiled with the following additive treatments: (1) control (i.e. no treatment), (2) formic acid‐based additive, (3) homofermentative lactic acid bacterial (LAB) inoculant and (4) heterofermentative LAB inoculant. Additive application to the PRG (irrespective of harvest date), the late‐harvest timothy (Harvest 3) and the red clover (Harvests 2 and 3) herbages had little impact on silage fermentation characteristics, where a lactic acid dominant fermentation already prevailed. However, the application of the formic acid‐based additive to the Harvest 1 and 2 timothy and Harvest 1 red clover herbages, where the silage fermentation characteristics were poorer, resulted in an increase in the proportion of lactic acid in total fermentation products and a decrease in pH. In contrast, the heterofermentative LAB inoculant had a negative impact on silage fermentation characteristics where a poor preservation prevailed in the corresponding control silage.     

On the Effect of Lactic Acid on Blood Glucose and Insulin Responses to Cereal Products: Mechanistic Studies in Healthy Subjects and In Vitro

It has been observed that bread containing lactic acid produced during the sourdough fermentation or added directly, has the ability to lower the postprandial glucose and insulin responses in humans. The main objective of the present work was to evaluate the possible mechanisms for a lowered glucose response to bread containing lactic acid, and to determine whether the same phenomenon also occurs when lactic acid is added to other cereal products. The rate of starch hydrolysis in bread and bread-like products was studied using an in vitro enzymatic approach. In addition, blood glucose and insulin responses to different lactic acid fermented barley gruels were evaluated in healthy subjects. It was concluded that the inclusion of lactic acid in bread reduces the rate of starch digestion by creating interactions between the gluten and starch. The presence of lactic acid during starch gelatinisation appeared to be a prerequisite for a reduced starch bioavailability. No effect of lactic acid was seen in gruels where the acid was formed after heat-treatment.     

Fermentation-induced changes in the nutritional value of native or germinated rye

Processing is important both for the sensory and nutritional quality of grains. In the case of whole grains, it especially may influence the levels and bioavailability of the bioactive compounds. Yeast, lactic acid and mixed fermentation were shown to vary in their effects on native and germinated wholemeal rye flour. The presence of Saccharomyces cerevisiae was important for enhanced levels of the analysed compounds. The levels of folates, free phenolic acids, total phenolic compounds, lignans and alkylresorcinols were increased especially by fermentation of germinated rye, which also resulted in lower pH when compared with native rye flour, the level of folate was increased up to seven-fold and that of free phenolic acids by up to ten-fold after germination and fermentation. Tailored fermentation thus offers a tool to further increase the bioactive potential of wholemeal rye.     

Influence of maceration at cutting on lactic acid bacteria populations, silage fermentation and voluntary intake and digestibility of precision-chopped lucerne silage

Laboratory experiments with lucerne (Medicago sativa) have shown that maceration at cutting improves silage fermentation. Samples taken during wilting and after various ensiling periods were analysed for lactic acid bacteria (LAB) numbers and indices of silage fermentation. In Experiment 1, in which maceration was tested in unwilted and wilted lucerne, there was an additive effect of maceration and wilting on LAB numbers at ensiling, thus LAB numbers were approximately 108 colony-forming units (cfu) g^?1 fresh crop for wilted, macerated forage compared with 103 cfu g^?1 fresh crop for unwilted, unmacerated forage at ensiling. Initially, maceration reduced pH (P < 0·001) and increased lactic acid (unwilted comparison only; P < 0·001) and insoluble N (wilted comparison only; P < 0·001) concentrations. After 70 d ensiling, beneficial effects of maceration were associated only with the wilted silage. In Experiment 2, macerated lucerne was compared with unmacerated material, which was either ensiled after a wilting period of similar length or after wilting had proceeded to the same DM concentration as in the macerated forage. During wilting, LAB numbers were significantly higher in macerated than unmacerated forage (P < 0·05). This was also the case during the first 16 h of ensiling (P < 0·01). A decline in pH was observed earlier in macerated silage. Two days after ensiling, lactic acid concentration was higher in macerated silage (P < 0·05), but insoluble N concentration was not different. In a third experiment, unconditioned forage was compared with forages ensiled after regular conditioning or maceration. Although drying rate over 30 h was not influenced by degree of conditioning, LAB numbers during wilting increased with the degree of conditioning. In silages made from these treatments after 6 h wilting, there were no major effects on fermentation characteristics. In a fourth experiment, digestibility and voluntary intake of precision-chopped silage were measured in sheep and found not to be increased by maceration. It was concluded that maceration per se resulted in marginal improvements in fermentation; however, when maceration also increased DM concentration, fermentation was markedly improved. In these precision-chopped silages, maceration had no effect on intake or digestibility.     

A model of clostridial dominance in ensilage

A simulation model of the growth and fermentation of lactic acid bacteria and clostridia in silage is described. The model is used to predict the occurrence of a well-preserved lactate silage or a poorly-preserved clostridial silage. Results of the model are compared with published experimental studies not used in the model's development. The results are summarized with maps showing the combinations of initial water-soluble-carbohydrate content, crop buffering capacity, dry matter content, temperature, initial pH, and initial population of lactic acid bacteria which result in a clostridial silage. The specific processes modelled include the growth and death of lactic acid bacteria and clostridia; the use of substrate and accumulation of fermentation end products; release of ammonia; and change in silage pH.