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1.
【目的】探究青藏高原垂穗披碱草青贮饲料中乳酸菌的多样性,筛选在低温条件下(10、15和25°C)生长性能较好的优良菌株。【方法】将垂穗披碱草青贮饲料中分离纯化的乳酸菌进行形态特征观察及16S r RNA基因测序鉴定;用MRS液体培养基在10、15和25°C条件下分离、初筛乳酸菌,选取高吸光度值的菌株作为优势菌株。用绿汁发酵液在10、15和25°C条件下培养测定其p H值,选取低p H值菌株作为优势菌株,并综合MRS培养基筛选结果确定优良菌株。【结果】从不同温度和发酵阶段的垂穗披碱草青贮饲料中共分离得到108个乳酸菌菌株,它们分属于6个属、18个种。其中,清酒乳杆菌LS-24在15°C条件下发酵液p H值显著降低(P0.05),戊糖片球菌PP-63在发酵初期生长速度较快,植物乳杆菌LP-21在15°C条件下发酵液p H值降至3.9且有最大活菌数。【结论】在青藏高原垂穗披碱草青贮饲料中发现的乳酸菌属基本涵盖了前人在常温青贮饲料中发现的所有属,但种数略少;在108株菌中,清酒乳杆菌LS-24、戊糖片球菌PP-63和植物乳杆菌LP-21在低温条件下均表现出较好的繁殖和发酵特性,可作为青贮饲料低温发酵的备选菌株。  相似文献   

2.
NaCl-tolerant lactic acid bacteria (LAB) strains LC-10 ( Lactobacillus casei ) and LP-15 ( Lact. plantarum ) and NaCl were used as additives to sorghun ( Sorghum bicolor ). Numbers of LAB were significantly ( P < 0·05) higher in all the additive-treated silages than in the control silage at an early stage of ensiling. During the fermentation process, addition of NaCl or LAB effectively inhibited the growth of aerobic bacteria and clostridia, but not yeasts. All the additive-treated silages had significantly ( P < 0·05) lower pH, ammonia nitrogen content, dry matter loss and gas production but significantly ( P < 0·05) higher lactic acid content and residual water soluble carbohydrates compared with the control silage. The improvement in silage quality was in the order : LAB > NaCl > control. Yeast counts were high in all additive-based silages and they increased during the exposure of the silages to air. As a result, these silages suffered aerobic deterioration, whereas the control silage was stable. The results confirmed that the NaCl or LAB improved fermentation quality but did not prevent aerobic deterioration of the silage.  相似文献   

3.
The effects of adding two legumes, Gliricidia sepium and Leucaena leucocephala, cv. Cunningham, and molasses on the fermentation characteristics of silages made from two tropical grasses (Pangola grass, Digitaria decumbens, and Setaria sphacelata cv. Kazungula) were investigated. Pangola grass silages contained significantly higher contents of water-soluble carbohydrates and lactic acid than did setaria silages after 100 days fermentation, but there were no significant differences between the two silages in populations of lactic acid bacteria and contents of total N and NH3–N. Addition of either species of legume had no significant effect on fermentation acids and NH3–N contents, and numbers of lactic acid bacteria. Addition of both legumes reduced NH3–N production in the silages by 59% after 5 days' fermentation. Numbers of lactic acid bacteria were not significantly affected by the different treatments. Enterococcus faecalis represented 60% of the lactic acid bacteria isolated from the treated herbages prior to ensiling. By 100 days of fermentation, only lactobacilli were isolated: 82% homo-fermenters and 18% hetero-fermenters. Lactobacillus mesenteroides subsp. dextranicum was found only in the silage supplemented with 33% (w/w) legume. It was concluded that the low quality of tropical grasses used as feeds for ruminants may be significantly improved by ensiling these grasses with small amounts of molasses and with high-protein tree leaves.M. Tjandraatmadja and B.W. Norton are with the Department of Agriculture. The University of Queensland, Queensland, 4072, Australia; I.C. Mac Rae is with the Department of Microbiology, The University of Queensland, Queensland, 4072, Australia.  相似文献   

4.
Degradation of grass fructans by epiphytic or inoculated lactic acid bacteria during ensilage was examined using both normal and sterile hybrid ryegrass. It was clear that even in the absence of bacteria fructan degradation occurred, but at a significantly slower rate than in normal grass which had not been inoculated with lactic acid bacteria. Fructan degradation in sterile herbage suggests that plant fructan hydrolases were partially responsible for this process in all herbages, irrespective of treatment. Inoculation of sterile herbage with a strain of Lactobacillus plantarum known to lack the ability to degrade grass fructans resulted in a slower rate of fructan breakdown than when inoculated with Lactobacillus casei subsp. paracasei , a confirmed fructan degrader. In the later stages of the fermentation of uninoculated normal herbage when water-soluble carbohydrate appeared to be limiting, lactic acid was fermented to acetic acid. However, this fermentation pathway was not observed in either of the inoculated silages. The results suggest that silage inoculant bacteria possessing fructan hydrolase activity may have potential for improving silage fermentation, particularly when late cut, low sugar grass containing a high proportion of fructans is ensiled.  相似文献   

5.
The fermentation of lactic acid by two Propionibacterium spp. and by Micrococcus lactilyticus was examined in simulated silage. Fermentation occurred in silages which achieved a low pH (<4·0) but this resulted only in a small rise in pH and a secondary clostridial fermentation did not occur. Lactic acid-fermenting organisms resembling Propionibacterium spp. were isolated from authentic silages but not from fresh grass.  相似文献   

6.
A natural lactic fermentation of mixtures of water and whole flour of either maize or high-tannin sorghum was obtained either before or after cooking to a weaning gruel: The preparations had a final pH of about 3.8 (range 3.67 to 4.00) and a ratio of lactic acid to acetic acid of 91 (w/w). The growth of added (about 107 c.f.u./g gruel) Gram-negative intestinal pathogenic bacteria, enterotoxigenicEscherichia coli, Campylobacter jejuni, Shigella flexneri andSalmonella typhimurium, was strongly inhibited in the sour gruels, and the effect could primarily be explained by the low pH caused by the formation of lactic and acetic acids during the fermentation process. Of the added Gram-positive bacteria,Bacillus cereus andStaphylococcus aureus showed similar inhibited growth up to 7h after inoculation in the sour gruels. The strain ofStaphylococcus, however, showed only a continued reduction in growth in the fermented gruel samples, which had a viable lactic bacteria culture indicating the presence of a bacteriocin. This implies that a low pH (< 4.0) alone is not sufficient to sustain the inhibition of the growth ofStaphylococcus aureus. The survival studies were carried out at optimal temperatures for each respective enteropathogen.  相似文献   

7.
The present study was aimed to investigate the nutritive profiles, microbial counts and fermentation metabolites in rye, Italian rye-grass (IRG) and barley supplemented with Lactobacillus plantarum under the field condition, and its probiotic properties. After preparation of silage, the content of crude protein (CP), crude ash, acid detergent fiber (ADF), and neutral detergent fiber (NDF), microbes such as lactic acid bacteria (LAB), yeast and fungi counts, and fermentation metabolites lactic acid, acetic acid and butyric acid was assessed. Results indicated that the content of ADF and NDF were significantly varied between rye, IRG and barley mediated silages. The content of CP was increased in L. plantarum supplemented with IRG, but slightly decreased in rye and barley mediated silages. The maximum LAB count was recorded at 53.10 × 107 cfu/g in rye, 16.18 × 107 cfu/g in IRG and 2.63 × 107 cfu/g in barley silages respectively. A considerable number of the yeasts were observed in the IRG silages than the rye silages (P < 0.05). The amount of lactic acid production is higher in L. plantarum supplemented silages as compared with control samples (P < 0.05). It was confirmed that higher amount of lactic acid produced only due to more number of LAB found in the silages. L. plantarum was able to survive at low pH and bile salt and the duodenum passage with the highest percentage of hydrophobicity. Furthermore, the strain was sensitive towards the antibiotics commonly used to maintain the microbes in food industrial setups. In conclusion, supplementation of L. plantarum is most beneficial in rye, IRG and barley silage preparations and probiotic characteristics of L. plantarum was an intrinsic feature for the application in the preparation of animal feeds and functional foods.  相似文献   

8.
Whole-crop wheat and barley were each harvested at the soft-, medium- and hard-dough stages of grain development. Material from each harvest was ensiled in polythene bag silos without additive or after the addition of calcium hydroxide, sodium hydroxide or sodium acrylate at 50, 50 and 12.5 g kg?1 of the crop dry matter (DM), respectively. All silages were opened after 60 days.With advancing maturity there was an increase in the content of DM, starch and insoluble-nitrogen, but a reduction in water-soluble carbohydrates (WSC) and ash.When crops were ensiled without additives, only medium-dough barley fermented to give butyric acid. However, the addition of calcium hydroxide to crops of low DM (soft-dough) and medium DM (medium-dough) promoted the activity of clostridial bacteria giving rise to the production of butyric acid, but this did not occur with crops of high DM (hard-dough). Sodium hydroxide gave rise to butyric acid only at low DM, and to restricted fermentation at high DM content. Sodium acrylate restricted fermentation and prevented butyric acid production in all silages.Ensiling led to an average reduction of 5 percentage units in the digestible organic matter (DOM) of the control silages compared to that of the crops. Addition of calcium hydroxide and sodium acrylate gave values similar to the control silages. Only sodium hydroxide consistently increased DOM, the effect becoming more marked as the crops matured. The increases over the control silages were 10, 18 and 26 units for wheat and 15, 21 and 20 units for barley at low, medium and high DM, respectively.  相似文献   

9.
The effect of applying a commercial lactic acid bacterial inoculant, at 5.6 × 104 cfu/g fresh material, to vetch, wheat, direct-cut and wilted alfalfa silages has been studied under laboratory conditions, and on wheat also under farm conditions. Dry matter losses in the inoculated vetch and alfalfa silages were smaller than in the control silages, due to improved fermentation in the former as indicated by a faster and larger pH decrease and by a faster and larger lactic acid build-up. Volatile fatty acid analysis also indicated more efficient fermentation patterns in the inoculated vetch and alfalfa silages with less ethanol, acetic and butyric acids compared with the respective control silages. The inoculant suppressed enterobacteria and clostridia in the inoculated direct-cut alfalfa silage. The inoculant did not have a great effect on the wheat silages.  相似文献   

10.
Silages are important feedstuffs. Homofermentative lactic acid bacterial inoculants are often used to control silage fermentation. However, some research pointed out those homofermentative lactic acid bacteria (LAB) impaired the aerobic stability of wheat, sorghum, and corn silages. Adding heterofermentative LAB can produce more acetic acid, thereby stabilizing silages during aerobic exposure. Alfalfa is difficult to ensile. The present work was to study the effects of L. buchneri (heterofermentative LAB), alone or in combination with L. plantarum (homofermentative LAB) on the fermentation, aerobic stability, bacteria diversity and ruminal degradability of alfalfa silage. After 90 days ensiling, the pH, NH3-N/TN, butyric acid content and molds counts of control were the highest. The inoculated silages had more lactic acid, acetic acid content and more lactic acid bacteria than the control. Inoculating LAB inhibited harmful microorganisms, such as Enterobacterium and Klebsiella pneumoniae. The L. buchneri L. plantarum-inoculated silage had more acetic acid and less yeasts than other three treatments (P < 0.05), and lower NH3-N/TN than control (< 0.05). The CO2 production of L. buchneri L. plantarum-inoculated silage was less than that of L. plantarum-inoculated silage (P < 0.05). Inoculating LAB in alfalfa silages can decrease pH, increase the production of lactic and acetic acids, reduce the number of yeasts and molds, and inhibit Enterobacterium and K. pneumoniae. Inoculating with L. buchneri or L. buchneri L. plantarum can improve aerobic stability of alfalfa silages. A combination of L. buchneri and L. plantarum is preferable because it enhanced alfalfa silage quality and aerobic stability.  相似文献   

11.
This study examined the production of com silages with low or high lactic acid concentrations, provided by the addition of formic acid (0.5%), molasses (5%) or microbial inoculant (homofermentative lactic acid bacteria, 10 g/tonne). After the fermentation period, sheep were fed the silages to determine true and apparent digestibility of the organic matter and microbial protein synthesis. The experiment were carried out with four KıvırcıkxMorkaraman sheep, 1.5 years old, fixed with cannula in their rumen and duodenum.Lactic acid concentrations were significantly higher in silages treated with enzyme or molasses compared to other specific treatments. Acetic acid concentration was highest in silage treated with formic acid, and lowest in silage treated with molasses (P < 0.05). The by-pass of crude protein was highest in silage treated with formic acid.  相似文献   

12.
AIMS: The detection and isolation of lactic acid bacteria by enrichment methods from wine grapes cultivated in vineyards located in New South Wales, Australia. METHODS AND RESULTS: Enrichment cultures in de Man, Rogosa and Sharpe (MRS) broth, MRS + ethanol (5%), MRS broth supplemented with 15% (v/v) tomato juice (MRST), pH 5.5 and 3.5 and autoenrichment in grape juice homogenate were used to detect lactic acid bacteria on wine grapes. Bacteria were isolated from enrichment cultures by plating onto MRS and MRST agar and identified by 16S rDNA sequence analysis and phenotypical methods. A molecular method, PCR-denaturing gradient gel electrophoresis (DGGE) was also used to examine the bacteria that developed in enrichment cultures. Species of Lactobacillus, Enterococcus, Lactococcus and Weissella were detected in enrichments by plating and PCR-DGGE. Other bacteria (Sporolactobacillus, Asaia, Bacillus ssp.) were also found in some enrichment cultures. The principal malolactic bacterium, Oenococcus oeni, was not isolated. CONCLUSIONS: The incidence and populations of lactic acid bacteria on wine grapes were very low. Damaged grape berries showed a greater presence of these bacteria than undamaged berries. The diversity of bacterial species isolated from the grapes was greater than those previously reported and represented both lactic acid bacteria and nonlactic acid bacteria. Some of these bacteria (i.e. Lactobacillus lindneri, Lactobacillus kunkeei) could be detrimental to wine production. Oenococcus oeni was not found on grapes, but its recovery could be obscured by overgrowth from other species. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactic acid bacteria are significant in wine production because they conduct the malolactic fermentation and cause stuck or sluggish alcoholic fermentation and wine spoilage. This study investigates wine grapes as a potential source of these bacteria.  相似文献   

13.
When molasses was added during ensilage of three tropical grasses [hamil grass (Panicum maximum cv. Hamil), pangola grass (Digitaria decumbens) and setaria (Setaria sphacelata cv. Kazungula)] the final pH, concentration of fermentation acids (except lactic acid) and NH3–N content were all similar after 100 days of incubation. Pangola grass silage had significantly higher lactic acid content (66 g/kg dry matter) than the other two. Adding either 4 or 8% (w/w) molasses reduced NH3–N, volatile fatty acid content and pH but increased lactic acid content in the final silages. Numbers of lactic acid bacteria remained approximately constant during the course of the fermentation, although large differences were noted in the species composition of the populations. At the time of ensiling, only Pediococcus spp. and Leuconostoc spp. were detected. By 5 days, the homo-fermentative population, notably Lactobacillus plantarum, dominated (43%) and remained dominant. Hetero-fermentative rods were only detected in the 100-day silage, where they represented 29% of the strains isolated. Homo-fermenters were more abundant in pangola (60%) and setaria (47%) silages than hamil (27%) silages. Homo-fermenter populations were lowest in the 12-week forage. Molasses additions increased homo-fermenter populations. Pangola grass gave the best quality silage but, since the water-soluble carbohydrate content in the grasses was insufficient to promote a strong lactic fermentation, the addition of 20 to 30 kg molasses/tonne should achieve satisfactory preservation.M. Tjandraatmadja and B.W. Norton are with the Department of Agriculture, The University of Queensland, Queensland, 4072, Australia; I.C. Mac Rae is with the Department of Microbiology, The University of Queensland, Queensland, 4072, Australia.  相似文献   

14.
Distribution of spermine in bacilli and lactic acid bacteria   总被引:1,自引:0,他引:1  
Obligate moderately thermophilic bacilli and obligate moderately thermoacidophilic bacilli contained spermine as the major polyamine in addition to putrescine and spermidine. The identity of spermine was confirmed by thin-layer chromatography and high-performance liquid chromatography before and after treatment with putrescine oxidase. Using these methods, thermospermine and spermine can be separated; thermospermine was not present in these organisms. On the other hand, various facultative thermophiles and mesophilic strains of the genus Bacillus, including alkalophiles and halophiles, lack spermine and other tetraamines. No spermine was detected in several strains of mesophilic or facultative slightly thermophilic lactic acid bacteria, Lactobacillus and Streptococcus.  相似文献   

15.
Abstract: Inoculants are used as silage additives to improve preservation efficiency and to enhance animal performance. In most commercially available inoculants, homofermentative lactic acid bacteria (LAB) have been used because they are fast and efficient producers of lactic acid, improving natural silage fermentation. Specific LAB inuculants may also have beneficial effects on animal performance even if there is no effect on fermentation. However, these types of inoculants are not always advantageous. They do not necessarily prevent sermentation by clostridia in moist silages, and they sometimes impair the aerobic stability of grass and small grain silages. Therefore, new criteria for silage inoculants should be established which consider the specific needs of the crop being ensiled. New approaches which are being taken to develop improved inoculants for silage include the following: (1) using LAB isolates which are more specific to the target crops; (2) inclusion of heterofermentative LAB to produce volatile fatty acids to inhibit yeasts and moulds upon aerobic exposure; (3) inclusion of organisms other than LAB in inoculants to inhibit detrimental microorganisms; (4) selection or engineering of LAB strains to inhibit specific microorganisms; and (5) cloning and expression of genes which would enable selected LAB strains to utilize polysaccharides in crops which are low in soluble carbohydrates. Many of these new strategies for formulating inoculants are being tested, but further research is needed to determine the most successful approaches.  相似文献   

16.
Twenty-four low dry matter (DM) silages differing in fermentation quality were harvested at the same time from a crop that consisted mainly of timothy (Phleum pratense), and meadow fescue (Festuca pratensis). The silage samples were analysed by gas chromatography (GC) - mass spectrometry and gas chromatography - flame ionisation detection in order to determine and quantify volatiles present in silage. The voluntary intake of the 24 silages had been measured in a previous feeding trial with growing steers of Norwegian Red. Thirteen esters, five aldehydes, three alcohols, and one sulphide were identified and quantified. A total of 51 variables describing the chemical composition of the silages were included in a partial least-squares regression, and the relationship of silage fermentation quality to voluntary intake was elucidated. The importance of variables describing silage fermentation quality in relation to intake was judged from a best combination procedure, jack-knifing, and empirical correlations of the variables to intake. The GC-analysed compounds were mainly present in poorly fermented silages. However, compared with other explanatory chemical variables none of these compounds was of importance for the voluntary intake as evaluated by partial least-squares regression. A validated variance of 71% in silage DM intake was explained with the selected variables: total acids (TA), total volatile fatty acids (TVFA), lactic acid/total acid ratio and propionic acid. In this study extent (by the variable TA) and type of silage fermentation (by TVFA) influenced intake. Further, it is suggested that by restricting the fermentation in low DM grass silages the potential intake of silage DM is maximised.  相似文献   

17.
Aims: To determine the effects of wilting, storage period and bacterial inoculant on the bacterial community and ensiling fermentation of guinea grass silage. Methods and Results: Fermentation products, colony counts and denaturing gradient gel electrophoresis (DGGE) profiles were determined. There was more lactic acid than acetic acid in all silages, but the lactic acid to acetic acid ratio decreased with storage time. This shift from lactic to acetic acid was not prevented even with a combination of wilting and bacterial inoculant. The DGGE analyses suggest that facultatively heterofermentative lactic acid bacteria (Lactobacillus plantarum, Lactobacillus brevis and Lactobacillus pentosus) were involved in the shift to acetic acid fermentation. Conclusions: Lactic acid can dominate the fermentation in tropical grass silage with sufficient wilting prior to ensiling. Prolonged storage may lead to high levels of acetic acid without distinctive changes in the bacterial community. Significance and Impact of the Study: The bacterial community looks stable compared to fermentation products over the course of long storage periods in tropical grass silage. Acetic acid fermentation in tropical grass silage can be a result of the changes in bacterial metabolism rather than community structure.  相似文献   

18.

This study was conducted to evaluate the fermentative profile and microbial populations of wilted and non-wilted alfalfa silages ensiled with or without inoculant and the population dynamics of lactic acid bacteria (LAB) of wilted alfalfa plant and theirs silage. A 2?×?2?×?6 factorial arrangement was used, with the absence or presence of wilting (W), with and without bacterial inoculant (I) and six fermentation periods (P) (1, 3, 7, 14, 28 and 56 days), in a completely randomized design, with three replicates. The alfalfa was slightly wilted for 6 h and increased the dry matter content from 133.9 to 233.4 g/kg. It was performed the cultivation, followed by the isolation of LAB from samples of alfalfa forage before ensiling and its silage only in non-inoculated silages, after different fermentation periods. DNA was extracted from the isolated strains of LAB; the 16S rRNA gene sequences were amplified by PCR and the sequences were compared to those available from the GenBank database. Wilting provided silages with lower pH, ammonia nitrogen and acetic acid concentrations. The wilting process did not alter the amount of LAB; however, it affected the LAB diversity of the silages. The Lactobacillus plantarum was the predominant species in non-wilted and wilted silages.

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19.
Aims: To understand the effects of lactic acid bacteria (LAB) inoculation on fermentation products, aerobic stability and microbial communities of silage. Methods and Results: Wilted Italian ryegrass was stored in laboratory silos with and without inoculation of Lactobacillus rhamnosus and Lactobacillus buchneri. The silos were opened after 14, 56 and 120 days and then subjected to aerobic deterioration for 7 days. Intensive alcoholic fermentation was found in untreated silage; the sum of ethanol and 2,3‐butanediol content at day 14 was about 7 times higher than that of lactic and volatile fatty acids. Alcoholic fermentation was suppressed by L. rhamnosus and L. buchneri inoculation and lactic acid and acetic acid became the dominant fermentation products, respectively. Silages were deteriorated in untreated and L. rhamnosus‐inoculated silages, whereas no spoilage was found in L. buchneri‐inoculated silage. Enterobacteria such as Erwinia persicina, Pantoea agglomerans and Rahnella aquatilis were detected in untreated silage, whereas some of these bacteria disappeared or became faint with L. rhamnosus treatment. When silage was deteriorated, Lactobacillus brevis and Bacillus pumilus were observed in untreated and L. rhamnosus‐inoculated communities, respectively. The inoculated LAB species was detectable in addition to untreated bacterial communities. Saccharomyces cerevisiae and Pichia anomala were the main fungi in untreated and L. rhamnosus‐inoculated silages; however, P. anomala was not visibly seen in L. buchneri‐inoculated silage either at silo opening or after exposure to air. Conclusion: Inoculation with L. rhamnosus can suppress alcoholic fermentation of wilted grass silage with elimination of enterobacteria at the beginning of fermentation. Addition of L. buchneri may improve aerobic stability, with distinct inhibitory effect observed on P. anomala after silo opening. Significance and Impact of the Study: Bacterial and fungal community analyses help us to understand how inoculated LAB can function to improve the fermentation and aerobic stability of silage.  相似文献   

20.
The effects of temperature, aerobic and anaerobic conditions in the silo and plant characteristics [water-soluble carbohydrate (WSC) contents, growing season] on the fermentation characteristics of a tropical forage species, Sorghum bicolor cv. sugar-drip, were investigated. Silages fermented in oxygen-impermeable bags were well preserved and had low pH (3.7), high lactic acid [72 g kg–1 dry matter (DM) 80% of total acids], and low butyric acid (0.12 g kg–1 DM) and ammonia nitrogen (NH3–N) (57 g kg–1 total nitrogen contents. Conversely, the use of oxygen-permeable bags as silos allowed aerobic decomposition of the ensiled forages. Increasing the incubation temperature lowered the population of lactic acid bacteria, reduced lactic acid production and caused the pH to rise. The heterofermentative Leuconostoc spp. predominated on fresh forages but homofermentative Lactobacillus plantarum began to dominate after 5 and 8 days of fermentation. Heterofermentative lactobacilli, notably Lactobacillus brevis, were dominant among the isolates obtained from 100-day silages. Varying the WSC contents, by crushing and/or chopping the forage, and growing season did not significantly affect the fermentation quality of the resulting silages. It was concluded that the maintenance of anaerobic conditions is essential if good quality silage is to be produced from tropical forage species.  相似文献   

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