Characterization and application of lactic acid bacteria for tropical silage preparation

Strains TH 14, TH 21 and TH 64 were isolated from tropical silages, namely corn stover, sugar cane top and rice straw, respectively, prepared in Thailand. These strains were selected by low pH growth range and high lactic acid‐producing ability, similar to some commercial inoculants. Based on the analysis of 16S ribosomal RNA gene sequence and DNA‐DNA relatedness, strain TH 14 was identified as Lactobacillus casei, and strains TH 21 and TH 64 were identified as L. plantarum. Strains TH 14, TH 21, TH 64 and two commercial inoculants, CH (L. plantarum) and SN (L. rhamnosus), were used as additives to fresh and wilted purple Guinea and sorghum silages prepared using a small‐scale fermentation method. The number of epiphytic lactic acid bacteria (LAB) in the forages before ensilage was relatively low but the numbers of coliform and aerobic bacteria were higher. Sorghum silages at 30 days of fermentation were all well preserved with low pH (3.56) and high lactic acid production (72.86 g/kg dry matter). Purple Guinea silage inoculated with LAB exhibited reduced count levels of aerobic and coliform bacteria, lower pH, butyric acid and ammonia nitrogen and increased lactic acid concentration, compared with the control. Strain TH 14 more effectively improved lactic acid production compared with inoculants and other strains. © 2016 Japanese Society of Animal Science     

Effect of grape pomace on fermentation quality and aerobic stability of sweet sorghum silage

The objective of this study was to evaluate the effect of grape pomace (GP) with different adding levels (0%, 5%, 10% and 15%, fresh matter basis), alone (GP‐LAB) or in combine with an inoculant LAB (GP+LAB), on the fermentation quality and aerobic stability of sweet sorghum silage. After 90 days of ensiling in vacuumized mini‐silos, silages were subject to a 7‐day aerobic stability test, in which chemical, microbial and polyphenol composition were measured. In the GP‐LAB group, adding GP decreased (P < 0.05) concentrations of water‐soluble carbohydrate (WSC) and butyric acid in silage. In the GP+LAB group, adding GP increased (P < 0.05) concentrations of lactic acid, WSC and crude protein, decreased (P < 0.05) final pH value, NH₃‐N ratio and butyric acid concentration in silage. Polyphenol level was reduced (P < 0.05) after silage fermentation. During aerobic exposure, the fungi count, pH value and silage temperature increased (P < 0.05), the levels of lactic acid, acetic acid and polyphenols (quercetin 3‐O‐glucoside and quercetin 3‐O‐glucuronid) decreased (P < 0.05) in silage. GP+LAB treated silage had a lag phase for aerobic spoilage. When the fermentation products, microbial counts, chemical and polyphenol composition were considered, the use of 10% GP+LAB at ensiling could provide a valuable source for improved fermentation quality and aerobic stability of sweet sorghum silage.     

5株乳酸菌复合物与CaCO3,酶及尿素不同组合对全株玉米青贮品质影响

为探讨前期筛选出的5株对好氧细菌、酵母菌和霉菌具有优良抑菌活性乳酸菌的复合物及其与CaCO₃、酶及尿素不同组合对全株玉米青贮发酵品质及营养成分的影响,进一步筛选优质复合青贮生物制剂,共设置7个不同组合添加物进行全株玉米青贮,并设空白对照组,测定青贮3、10和30 d时其发酵品质和营养成分的变化情况。结果表明,添加5株乳酸菌混合剂有提高全株玉米青贮发酵品质和营养品质的趋势;同时添加CaCO₃和复合乳酸菌能进一步增加青贮饲料的乳酸(LA)和乙酸(AA)产量,尤其是AA产量,防止pH值过分降低,也有提高青贮营养品质的趋势;同时添加尿素和复合乳酸菌能进一步提高青贮的AA和氨态氮的含量,防止pH值过分降低,显著提高粗蛋白(CP)含量,从而提高青贮营养品质;纤维素酶和淀粉酶复合物可以提高发酵产物的水溶性碳水化合物(WSC)含量及LA和AA产量,改善青贮发酵品质;复合CaCO₃、尿素、纤维素酶、淀粉酶和复合乳酸菌的添加对提高全株玉米青贮的发酵品质和营养品质的效果最好;全株玉米青贮发酵品质和营养品质在前3 d变化最快,发酵不同阶段不同添加组合的变化不尽一致。     

Fermentation quality and aerobic stability of mulberry silage prepared with lactic acid bacteria and propionic acid

The objective of this study was to examine the fermentation quality and aerobic stability of mulberry (Morus alba L.) silage prepared with lactic acid bacteria (LAB) and propionic acid (PA). The selected LAB strains Lactobacillus (L.) plantarum LC365281 (L1) and L. brevis LC365282 (L2), and commercial inoculant strains L. plantarum Gaofuji (GF) and L. buchneri Fresh (FR), and PA were used as additives for silage preparation. Silage treatments were designed as control, L1, L2, GF, FR, PA, PA + L1, PA + L2, PA + GF, or PA + FR. After 30 days of ensiling, the fermentation quality of silages treated with PA + L1 was improved, with a lower (p < 0.05) pH and NH₃‐N content than those of other treatments. During the aerobic exposure, the PA + LAB‐treated silages displayed an aerobic stability with stable pH value and lactic acid content. The results confirm that L. plantarum L1 and PA were the best additive combination for ensiling mulberry.     

糖蜜和乳酸菌对去穗玉米秸秆青贮发酵品质的影响

为研究糖蜜和乳酸菌对去穗玉米（Zea mays L.）秸秆青贮饲料发酵品质的影响,试验设对照（C）、糖蜜（M）、乳酸菌（LAB）、糖蜜+乳酸菌（M+LAB）4个处理,发酵60天后测定发酵品质。结果表明：添加糖蜜或乳酸菌均提高了去穗玉米秸秆青贮饲料的发酵品质,M+LAB组的V-Score评分最高。因此,在去穗玉米秸秆青贮实际生产过程中,建议使用糖蜜和乳酸菌组合添加剂。     

A survey of fermentation products and bacterial communities in corn silage produced in a bunker silo in China

To evaluate the current practice of corn silage management in China, samples of bunker‐made silage were collected from 14 farms within a 500‐km radius of Beijing for the analysis of fermentation products and bacterial communities. Mean values for dry matter (DM) content were as low as 250 g/kg in both corn stover (St) and whole crop corn (Wc) silages, and pH values averaged 4.48 and 3.73, respectively. Only three of the 14 silages exhibited a lactic‐to‐acetic acid ratio > 1.0, indicating that the presence of acetic acid was predominant in fermentation. Although 1,2‐propanediol content was marginal in most cases (< 5.0 g/kg dry matter (DM)), two Wc silages had 1,2‐propanediol levels > 25 g/kg DM. In contrast, 3 St silages had large amounts (> 10 g/kg DM) of butyric acid, and two of the three butyrate silages also had high concentrations of 1‐propanol. Denaturing gradient gel electrophoresis analysis demonstrated that the bacterial community appeared similar in 10 out of the 14 silage samples. Bands indicating Lactobacillus buchneri, L. acetotolerans and Acetobacter pasteurianus were found in both the St and Wc silages, accounting for the high acetic acid content found across silage samples.     

不同乳酸菌添加剂对青贮黑麦草和青贮玉米发酵产物和有氧稳定性的影响

在青贮黑麦草(Lolium perenne)和青贮玉米(Zea mays)中添加鼠李糖乳杆菌和布氏乳杆菌,研究其对发酵产物与有氧稳定性的影响.青贮饲料贮藏时间为14,56和120 d.无添加青贮黑麦草发酵产物中乙醇含量较多,在14 d开封时,乙醇和2,3-丁二醇含量为乳酸和挥发性脂肪酸总和的7倍.添加鼠李糖乳杆菌与布氏乳杆菌可以抑制乙醇型发酵,使乳酸型与乙酸型发酵占主导.无添加青贮黑麦草与添加鼠李糖乳杆菌的黑麦草发生了有氧腐败,但是添加布氏乳杆菌的黑麦草保持了有氧稳定.在青贮玉米中,添加鼠李糖乳杆菌未影响发酵品质,但添加布氏乳杆菌增加了发酵产物中乙酸含量,降低了乳酸含量.添加布氏乳杆菌的青贮玉米从56 d开封开始保持了有氧稳定,其它组在120 d开封时保持了有氧稳定,所有组乙酸含量都增加.     

Silage fermentation and ruminal degradation of stylo prepared with lactic acid bacteria and cellulase

In order to improve the silage fermentation of stylo (Stylosanthes guianensis ) in tropical areas, stylo silages were prepared with commercial additives Lactobacillus plantarum Chikuso‐1 (CH 1), L. rhamnasus Snow Lact L (SN ), Acremonium cellulase (CE ) and their combination as SN +CE or CH 1 + CE , and the fermentation quality, chemical composition and ruminal degradation of these silages were studied. Stylo silages treated with lactic acid bacteria (LAB ) or cellulase, the pH value and NH ₃‐N ? total‐N were significantly (P  <  0.05) decreased while the ruminal degradability of dry matter (DM ), crude protein (CP ), neutral detergent fiber (aNDF om) and acid detergent fiber (ADF om) were significantly (P  <  0.05) increased compared to control. Compared to LAB or cellulase‐treated silages, the DM , CP contents and relative feed value (RFV ), and the ruminal degradability in LAB plus cellulase‐treated silages were significantly (P  <  0.05) higher, but the aNDF om content was significantly (P  <  0.05) lower. CH 1 + CE treatment was more effective in silage fermentation and ruminal degradation than SN +CE treatment. The results confirmed that LAB or LAB plus cellulase treatment could improve the fermentation quality, chemical composition and ruminal degradation of stylo silage. Moreover, the combined treatment with LAB and cellulase may have beneficial synergistic effects on ruminal degradation.     

5株优良抑菌活性乳酸菌对全株玉米青贮品质的影响

为了探讨具有优良抑菌活性的乳酸菌对全株玉米青贮过程不同时期营养品质的影响,进一步验证并筛选出可提高玉米秸秆青贮品质的乳酸菌接种剂,将实验室前期从甘肃各地玉米秸秆青贮饲料中分离筛选获得的5株产酸快、多,且具有抑菌活性的优良乳酸菌分别添加到全株玉米秸秆中进行青贮,共设置6个试验组,分别在青贮第3、7、15、30天测定产物的干物质(DM)、粗蛋白(CP)、可溶性碳水化合物(WSC)、中性洗涤纤维(NDF)、酸性洗涤纤维(ADF)和氨态氮(NH₃-N)含量及第30 天乳酸(LA)、乙酸(AA)、丙酸(PA)和丁酸(BA)含量。结果显示,发酵30 d后,5个处理组的DM、CP、ADF和LA含量显著高于对照组(P<0.05),NH₃-N和WSC含量显著低于对照组(P<0.05),B3-1组NDF显著高于对照组(P<0.05),但与其他处理组差异不显著(P>0.05),B3-1和B5-2组AA含量显著高于对照组(P<0.05),但与其他处理组差异不显著(P>0.05);各组DM、CP、WSC含量均随发酵时间延长而降低,而ADF、NDF和NH₃-N含量均随发酵时间的延长而升高;B3-1组在玉米秸秆全株青贮发酵各期的发酵品质和营养品质最优。以上结果表明5株乳酸菌均能显著改善青贮饲料发酵品质,其中植物乳杆菌B3-1的效果最好。     

Effect of lactic acid bacteria,molasses, and their combination on the fermentation quality and bacterial community of cassava foliage silage

This study investigated the effects of LAB inoculants (L) and molasses (M) on the microbial community and fermentation quality of cassava foliage (CF). The small segments (about 2–3 cm) CF were ensiled in plastic bags and incubated at normal temperature (25°C). Four treatments were carried out as follows: control (no additives, CK), LAB inoculants (Lactobacillus plantarum, L), molasses (M), and LAB in combination with molasses (LM). The LAB and molasses obviously altered the bacterial community structure of the CF silage and enhanced the fermentation quality. The combination addition could increase the abundance of Lactobacillus and reduce the Pseudomonas. The LAB and molasses also significantly elevated the lactic acid concentration (P < 0.001) and decreased the pH (P < 0.001), as well as the concentrations of acetic acid, propionic acid, butyric acid, and ammonia-N (P < 0.05). In addition, the combination treatment displayed more effective results on silage fermentation. The LAB and molasses improved the fermentation quality of the CF silage by altering the bacterial community structure. Furthermore, the bacterial community was significantly correlated with fermentation indexes.     

Characteristics of isolated lactic acid bacteria and their effectiveness to improve stylo (Stylosanthes guianensis Sw.) silage quality at various temperatures

Two lactic acid bacteria (LAB) strains, Pediococcus pentosaceus SC1 and Lactobacillus paraplantarum SC2 isolated from king grass silage, were characterized and their effectiveness to improve the silage fermentation quality of stylo (Stylosanthes guianensis Sw.) was studied. Strain SC1 was able to grow at a high temperature of 45°C, while SC2 did not. SC2 normally grew at a low pH of 4.0, while SC1 could not. These two strains and a commercial inoculant of LAB (L. plantarum, LP) were used as additives to stylo silage preparation at various temperatures (20°C, 30°C and 40°C). All LAB inoculants significantly (P < 0.05) reduced the pH value and ammonia-N content, and increased the ratio of lactic acid to acetic acid and quality score compared with the control. In addition, inoculating LAB strains markedly (P < 0.05) reduced butyric acid content at the temperatures of 30°C and 40°C. Compared to SC2 and LP strains, strain SC1 was the most effective for improving stylo silage quality at 20°C, indicated by the increase in lactic acid, ratio of lactic acid to acetic acid and quality score. At 30°C and 40°C, there were no significant differences among SC1, SC2 and LP treatments in pH values, contents of acetic acid, butyric acid and ammonia-N (P > 0.05).     

Fermentation quality and chemical composition of shrub silage treated with lactic acid bacteria inoculants and cellulase additives

Effects of lactic acid bacteria (LAB) inoculants and cellulase additives on fermentation quality and chemical compositions of shrub silages were studied by using a small‐scale fermentation system. Two LAB inoculants of Qingbao (Lactobacillus plantarum, Pediococcus acidilacticii, Lactobacillus casei and Clostridium phage) and Caihe (Lactobacillus plantarum, Lactobacillus brevis and Pediococcus acidilactici) and a commercial cellulase made from Trichoderma reesei were used as additives for intermediate pea‐shrub, rush bushclover, arborescent ceratoides and shrubby silage preparation. The crude protein, neutral detergent fiber and water‐soluble carbohydrate contents of the four shrub materials were 10.1–14.2, 62.6–67.2 and 1.9–3.5% on a dry matter basis, respectively. All shrub silages had pH 3.40–4.43, ammonia‐N 0.1–0.2% g/kg and lactic acid 1.3–2.9% on a fresh matter basis. The silage quality of LAB‐inoculated silages did not have a greater effect than control silages, except shrubby silage preparation. Silages treated with the cellulase, the pH of rush bushclover and shrubby sweetvetch silage were significantly (P < 0.05) lower and the lactic acid content were significantly (P < 0.05) higher than the control silages. The results confirmed that shrub contained a relatively high content of crude protein; its silages can be preserved in good quality, and they are new potential resources for livestock feed.     

Microbial population,chemical composition and silage fermentation of cassava residues

In order to effectively use the cassava (Manihot esculenta Crantz) residues, including cassava leaves, peel and pulp for livestock diets, the chemical and microbiological composition, silage preparation and the effects of lactic acid bacteria (LAB) inoculants on silage fermentation of cassava residues were studied. These residues contained 10⁴ to 10⁵ LAB and yeasts, 10³ to 10⁴ coliform bacteria and 10⁴ aerobic bacteria in colony forming units (cfu) on a fresh matter (FM) basis. The molds were consistently at or below the detectable level (10² cfu of FM) in three kinds of cassava residues. Dry matter (DM), crude protein (CP) and neutral detergent fiber (NDF) content of cassava residues were 17.50‐30.95%, 1.30‐16.41% and 25.40–52.90% on a DM basis, respectively. The silage treatments were designed as control silage without additive (CO) or with LAB inoculants Chikuso‐1 (CH, Lactobacillus plantarum) and Snow Lacto (SN, Lactobacillus rhamnosus) at a rate of 5 mg/kg of FM basis. All silages were well preserved with a low pH (below 4.0) value and when cassava residues silage treated with inoculants CH and SN improved fermentation quality with a lower pH, butyric acid and higher lactic acid than control silage.     

玉米粉和乳酸菌对甘薯蔓、酒糟及稻草混合青贮品质的影响

为评价玉米粉和乳酸菌对甘薯蔓、酒糟及稻草(4∶4∶2)混合青贮品质的影响,试验设对照组(CK)、玉米粉添加组(CF)、乳酸菌添加组(LAB)、玉米粉和乳酸菌组合添加组(CF+LAB),每个处理6个重复,室温下贮藏60 d开封,取样分析青贮品质。结果表明,添加玉米粉和乳酸菌制剂均明显提高了青贮料的感官品质,与CK相比,CF处理、LAB处理及CF+LAB处理中 CP含量极显著提高(P<0.01), NH₃-N/TN、AA、PA、Ash含量极显著降低(P<0.01),CF处理极显著地提高了DM、CP、LA含量(P<0.01),而LAB处理则极显著地降低了NDF、ADF含量(P<0.01),CF+LAB中NH₃-N/TN、AA含量及pH值极显著低于LAB(P<0.01),显著低于CF(P<0.05)。综上所述,添加玉米粉和乳酸菌制剂均提高了青贮品质,单独添加乳酸菌制剂青贮品质要次于单独添加玉米粉,两者组合添加青贮品质更好。     

玉米秸秆和白菜尾菜混贮料的乳酸菌多样性及耐高温优良菌株筛选

通过形态特征、生理生化特性及16S rRNA序列分析方法对玉米(Zea mays)秸秆和白菜(Brassica pekinensis)尾菜混贮料中的乳酸菌多样性进行分析,并以温度和p H为限制因素筛选优良乳酸菌菌株。结果表明,分离得到的12株乳酸菌分属于乳杆菌属(Lactobacillus)和片球菌属(Pediococcus)。其中,1株(LB-1)为植物乳杆菌(Lactobacillus plantarum),6株(LB-2、LB-4、LB-7、LB-8、LB-9和LB-11)为戊糖片球菌(Pediococcus pentosaceus),3株(LB-5、LB-6和LB-12)为短乳杆菌(Lactobacillus brevis),2株(LB-3和LB-10)为类干酪乳杆菌(Lactobacillus paracasei)。菌株LB-3和LB-8表现出优良的耐高温、耐酸碱特性,且具有较强的产乳酸能力,二者可作为青贮饲料的乳酸菌添加剂。     

Isolating and evaluating lactic acid bacteria strains for effectiveness on silage quality at low temperatures on the Tibetan Plateau

Four lactic acid bacteria (LAB) strains isolated from straw silages on the Tibetan Plateau were characterized, and their effects on the fermentation quality of Italian ryegrass (Lolium multiflorum Lam.) at different temperatures (10°C, 15°C and 25°C) were studied. These LAB isolates were evaluated using the acids production ability test, morphological observation, Gram staining, physiological, biochemical and acid tolerance tests. All the isolates (M1, LM8, LO7 and LOG9) could grow at 5‐20°C, pH 3.5‐7.0 and NaCl (3.0%, 6.5%). Strains M1, LM8, LO7 and LOG9 were identified as Lactobacillus plantarum, L. coryniformis, Pediococcus pentosaceus and P. acidilactici, respectively, by sequencing 16S ribosomal DNA. The four isolates were added to Italian ryegrass for ensiling for 30 days at various temperatures. Compared with the corresponding control, inoculating with isolates M1, LM8 and LO7 could improve the silage quality of Italian ryegrass at low temperatures, indicated by significantly (P < 0.05) higher lactic acid (LA) contents and ratios of lactic acid/acetic acid (LA/AA), and significantly (P < 0.05) lower pH and ammonia nitrogen/total nitrogen (AN/TN). Compared with other isolates, LM8 performed better at 10°C and 15°C, indicated by the higher (P < 0.05) LA content and ratio of LA/AA, and the lower (P < 0.05) pH and AN/TN.     

优良抑菌活性乳酸菌对玉米青贮及有氧暴露期微生物数量和pH的影响

为了探讨乳酸菌对全株玉米青贮及有氧暴露后青贮饲料中乳酸菌、好氧细菌、酵母菌和霉菌数量及其pH的影响,进一步筛选出可提高青贮饲料品质和有氧稳定性的乳酸菌接种剂,将实验室前期从甘肃各地玉米秸秆青贮饲料中分离筛选获得的5株产酸快、多且具有抑菌活性的优良乳酸菌分别添加全株玉米进行青贮,分析青贮过程和有氧暴露后青贮饲料中乳酸菌、好氧细菌、酵母菌和霉菌数量的动态变化及pH。结果显示,在青贮过程和有氧暴露后,分别添加肠膜明串珠菌肠膜亚种B1-7、戊糖片球菌B2-3、植物乳杆菌B3-1、屎肠球菌B5-2和发酵乳杆菌E2-3的各处理组乳酸菌总数均显著高于对照组,而好氧细菌、酵母菌和霉菌数量均显著低于对照组,pH亦低于对照组。其中B1-7和B5-2处理组在青贮初期乳酸菌总数最多,从青贮第7天开始到有氧暴露的30 d内,始终是B3-1处理组乳酸菌总数最多,好氧细菌、酵母菌和霉菌数量最少、pH最低。以上结果表明这5株乳酸菌具有提高青贮饲料品质和有氧稳定性的潜力,其中植物乳杆菌B3-1的效果最好。     

Effects of inoculants and environmental temperature on fermentation quality and bacterial diversity of alfalfa silage

To investigate the effects of inoculants and environmental temperature on fermentation quality and bacterial diversity of alfalfa silage, first‐cut alfalfa was ensiled with or without two screened lactic acid bacteria (LAB) strains, Lactobacillus plantarum, LP, and Lactobacillus casei, LC. Each treatment was divided into three parts and stored at 20°C, 30°C, 40°C, respectively. After 60 days ensiling, fermentation characteristics were measured and bacterial diversity was investigated by 16S ribosomal RNA gene sequencing using Illumina MiSeq platform. LP and LC decreased pH, coliform bacteria counts and increased lactic acid content at 20°C, and the two strains decreased pH, ammonia‐N concentration, coliform bacteria counts at 30°C. When the environmental temperature was 40°C, silage treated with LC showed lower LAB and coliform bacteria counts and higher lactic acid content than the untreated and LP treated silages. Butyric acid mainly appeared in silages stored at 40°C. The relative abundance of Lactobacillus in alfalfa silages stored at 20°C and 30°C was highest and increased after LP and LC were added. Garciella was another dominant genus in silages stored at 40°C. In conclusion, LP and LC improved fermentation quality of alfalfa silage by increasing Lactobacillus proportions at 20°C and 30°C; ensiling alfalfa at 40°C was difficult because of Garciella.     

Effect of total mixed ration silage containing agricultural by-products with the fermented juice of epiphytic lactic acid bacteria on rumen fermentation and nitrogen balance in ewes

The goal of this study was to determine the effects of the fermented juice of epiphytic lactic acid bacteria (FJLB) on the quality of total mixed ration (TMR) silage containing agricultural by-products, its digestibility, rumen fermentation, and nitrogen balance in ewes. TMR was prepared from rice straw, corn stover silage, brewer grain, tofu waste, steam-flaked corn, and a mineral mixture. The treatments consisted of silage additives added to TMR: CON (no silage additive), FJLB, COM (commercial additive), and MIX (FJLB + COM). Four cannulated ewes were assigned to the 4 × 4 Latin square design. The MIX treatment produced a lower (P < 0.01) pH than did the CON and FJLB treatments and a higher (P < 0.01) lactic acid concentration than did the other treatments. The fiber content in the COM treatment was lower (P < 0.05) than that in the other treatments. The FJLB treatment had similar fermentation quality and chemical composition to those of the CON and COM treatments in all parameters observed. Although the silage quality index (Fleig point) was higher in the MIX and COM treatments than in the CON treatment, all silages had good quality. No silage additives affected intake, digestibility, rumen fermentation, or nitrogen balance. In conclusion, the TMR silage prepared from agricultural by-products mixed with wet-type food by-products with or without FJLB added resulted in well-preserved fermentation, and this product might be used as a ruminant feed.

     

Fermentation and microbial population dynamics during the ensiling of native grass and subsequent exposure to air

To study the microbial population and fermentation dynamics of large needlegrass (LN) and Chinese leymus (CL) during ensiling and subsequent exposure to air, silages were sampled and analyzed using culture‐based techniques and denaturing gradient gel electrophoresis (DGGE). A total of 112 lactic acid bacteria (LAB) strains were isolated and identified using the 16S rRNA sequencing method. Lactic acid was not detected in the first 20 days in LN silage and the pH decreased to 6.13 after 45 days of ensiling. The temperature of the LN silage increased after approximately 30 h of air exposure and the CL silage showed a slight temperature variation. Enterococcus spp. were mainly present in LN silage. The proportion of Lactobacillus brevis in CL silage increased after exposure to air. LN silage with a higher proportion of Enterococcus spp. and propionic acid concentration did not show higher fermentation quality or aerobic stability than CL silage, which had a higher concentration of acetic acid, butyric acid and increased proportion of L. brevis after exposure to air.