Lactobacillus spp. from an inoculant andWeissella and Leuconostoc spp. from forage crops were characterized, and their influence on silage fermentation was studied. Forty-two lactic acid-producing cocci were obtained from forage crops and grasses. All isolates were gram-positive, catalase-negative cocci that produced gas from glucose, and produced more than 90% of their lactate in the d-isomer form. These isolates were divided into groups A and B by sugar fermentation patterns. Two representative strains from the two groups, FG 5 and FG 13, were assigned to the species Weissella paramesenteroides and Leuconostoc pseudomesenteroides, respectively, on the basis of DNA-DNA relatedness. Strains FG 5, FG 13, and SL 1 (Lactobacillus casei), isolated from a commercial inoculant, were used as additives to alfalfa and Italian ryegrass silage preparations. Lactic acid bacterium counts were higher in all additive-treated silages than in the control silage at an early stage of ensiling. During silage fermentation, inoculation with SL 1 more effectively inhibited the growth of aerobic bacteria and clostridia than inoculation with strain FG 5 or FG 13. SL 1-treated silages stored well. However, the control and FG 5- and FG 13-treated silages had a significantly (P < 0.05) higher pH and butyric acid and ammonia nitrogen contents and significantly (P < 0.05) lower lactate content than SL 1-treated silage. Compared with the control silage, SL 1 treatments reduced the proportion ofd-(−)-lactic acid, gas production, and dry matter loss in two kinds of silage, but the FG 5 and FG 13 treatments gave similar values in alfalfa silages and higher values (P < 0.05) in Italian ryegrass silage. The results confirmed that heterofermentative strains of W. paramesenteroides FG 5 andL. pseudomesenteroides FG 13 did not improve silage quality and may cause some fermentation loss.

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.

Pediococcus species isolated from forage crops were characterized, and their application to silage preparation was studied. Most isolates were distributed on forage crops at low frequency. These isolates could be divided into three (A, B, and C) groups by their sugar fermentation patterns. Strains LA 3, LA 35, and LS 5 are representative isolates from groups A, B, and C, respectively. Strains LA 3 and LA 35 had intragroup DNA homology values above 93.6%, showing that they belong to the species Pediococcus acidilactici. Strain LS 5 belonged to Pediococcus pentosaceus on the basis of DNA-DNA relatedness. All three of these strains and strain SL 1 (Lactobacillus casei, isolated from a commercial inoculant) were used as additives to alfalfa and Italian ryegrass silage preparation at two temperatures (25 and 48°C). When stored at 25°C, all of the inoculated silages were well preserved and exhibited significantly (P < 0.05) reduced fermentation losses compared to that of their control in alfalfa and Italian ryegrass silages. When stored at 48°C, silages inoculated with strains LA 3 and LA 35 were also well preserved, with a significantly (P < 0.05) lower pH, butyric acid and ammonia-nitrogen content, gas production, and dry matter loss and significantly (P < 0.05) higher lactate content than the control, but silages inoculated with LS 5 and SL 1 were of poor quality. P. acidilactici LA 3 and LA 35 are considered suitable as potential silage inoculants.

Enterococcus faecalis CA 6, E faecium CA 10, E casseliflavus CA 13, Weissella paramesenteroides CA 14, Leuconostoc pseudomesenteroides CA 17, Pediococcus pentosaceus CA 21, P acidilactici CA 25 and Lactobacilus plantarum CA 28 isolated from forage crops were characterised and their effects on silage fermentation were studied. Strains CA 6, CA 10 and CA 13 grew only at relatively high pH (above 5.0), while strains CA 21, CA 25 and CA 28 were able to grow at pH values below 4.0. Strain CA 25 grew at 50 °C, but the others did not grow at above 45 °C. These strains were used as inoculants for perennial ryegrass silage ensiled at 25 or 45 °C. At 25 °C all strains significantly (P < 0.01) reduced the pH value and butyric acid content and greatly increased the lactic acid content compared with the control, particularly strains CA 21, CA 25 and CA 28. At 45 °C strain CA 25 markedly improved the fermentation quality of silage, but the others did not. These silages had a lower lactic acid content and a higher proportion of L(+)‐ to total lactic acid than equivalent silages kept at 25 °C. The results confirmed that at 25 °C P pentosaceus CA 21, P acidilactici CA 25 and L plantarum CA 28 were the most effective, whereas at 45 °C only P acidilactici CA 25 had a beneficial effect on silage fermentation quality. © 2000 Society of Chemical Industry

Y . C AI , S. OH M OM O, M . O GA W A A ND S . K UM A I. 1997. 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 ³ 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 ³ 0·05) lower pH, ammonia nitrogen content, dry matter loss and gas production but significantly (P ³ 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.