Chinese Journal of Animal Nutrition >

2023 , Vol. 35 >Issue 4: 2702 - 2710

DOI: https://doi.org/10.12418/CJAN2023.252

RESEARCH PAPER

Effects of Different Lactic Acid Bacteria Treatments on Nutritional Quality, Fermentation Characteristics and Aerobic Stability of Hybrid Broussonetia papyrifera Silage

  • ZHANG Yulin , 1, 2 ,
  • YANG Hanjun 1 ,
  • LI Chaocheng 3 ,
  • QI Yayin 1 ,
  • MA Chunhui 1 ,
  • ZHANG Fanfan , 1, *
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  • 1 College of Animal Science and Technology, Shihezi University, Shihezi 832000, China
  • 2 College of Animal Science and Technology, Tarim University, Alar 843300, China
  • 3 China Xinjiang Kegou Biolgy Science and Technolgy Co., Ltd., Shihezi 832000, China
*associate professor, E-mail:

Received date: 2022-10-07

  Online published: 2023-04-12

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Abstract

This experiment was conducted to study the effects of different lactic acid bacteria treatments on nutritional quality, fermentation characteristics and aerobic stability of hybrid Broussonetia papyrifera silage, and to provide technical reference for the development and utilization of hybrid Broussonetia papyrifera. There were 7 treatments as follows: the CK treatment was supplemented 10 mL/kg deionized water, the LP treatment was supplemented 5×106 CFU/g FM Lactobacillus plantarum, the LB treatment was supplemented 5×106 CFU/g FM Lactobacillus brevis, the EF treatment was supplemented 5×106 CFU/g FM Enterococcus faecalis, the LP+EF treatment was compound supplemented 5×106 CFU/g FM Lactobacillus plantarum and Enterococcus faecalis (1∶1), the LP+LB treatment was compound supplemented 5×106 CFU/g FM Lactobacillus plantarum and Lactobacillus brevis (1∶1), and the LP+LB+EF treatment was compound supplemented 5×106 CFU/g FM Lactobacillus plantarum, Lactobacillus brevis and Enterococcus faecalis (1∶1∶1), each treatment contained 5 replicates, fermentation for 60 days. The results showed as follows: 1) the dry matter content of LP and LP+LB+EF treatments was significantly higher than that of CK treatment (P<0.05). The neutral detergent fiber content of LP, LB, EF, LP+LB and LP+LB+EF treatments was significantly lower than that of CK treatment (P<0.05). The crude protein content of EF and LP+LB+EF treatments was significantly higher than that of CK treatment (P<0.05). The water-soluble carbohydrates content of LP, LB and EF treatments was significantly lower than that of CK treatment (P<0.05). 2) The pH of LP and LP+LB+EF treatments was significantly lower than that of CK treatment (P<0.05). The ammoniacal nitrogen content of CK treatment was significantly higher than that of other treatments (P<0.05). The lactic acid content of LP treatment was significantly higher than that of EF, LP+LB and CK treatments (P<0.05). The acetic acid content of LB and LP+LB treatment was significantly higher than that of other treatments except LP+LB+EF treatment (P<0.05). The propionic acid content of LP, LB and LP+LB treatment was significantly lower than that of CK treatment (P<0.05). The lactic acid bacteria population of LP+LB treatment was significantly higher than that of other treatments (P<0.05). 3) After 5 days of aerobic exposure, the mold population of LP+LB treatment was the lowest, and the aerobic bacteria population of LP+LB+EF treatment was the lowest. The aerobic stability order from high to low was LB treatment > LP+LB treatment > LP+LB+EF treatment > CK, LP, EF and LP+EF treatments. To sum up, adding epiphytic lactic acid bacteria to hybrid Broussonetia papyrifera silage obviously improve the nutritional quality, fermentation characteristics and aerobic stability.

Key words： lactic acid bacteria; Broussonetia papyrifera; silage; nutritional quality; fermentation characteristics; aerobic stability

Cite this article

ZHANG Yulin , YANG Hanjun , LI Chaocheng , QI Yayin , MA Chunhui , ZHANG Fanfan . Effects of Different Lactic Acid Bacteria Treatments on Nutritional Quality, Fermentation Characteristics and Aerobic Stability of Hybrid Broussonetia papyrifera Silage[J]. Chinese Journal of Animal Nutrition, 2023 , 35(4) : 2702 -2710 . DOI: 10.12418/CJAN2023.252

杂交构树(Broussonetia papyrifera)是由中国科学院植物研究所通过杂交育种技术培育出的一种木本植物品种,其营养价值丰富、抗逆性强、生长速度快[1],用作饲料可提高各类家畜生长性能、肉品质等[2⇓-4]。因此,大力开发杂交构树资源对缓解我国蛋白质饲料短缺问题具有重要现实意义。由于杂交构树蛋白质分子结构复杂,直接鲜饲畜禽消化利用率较低,导致饲料资源的大量损失;而青贮发酵能减缓杂交构树中营养物质的流失,有效改善家畜适口性和消化率,相较于调制干草(粉)具有较好的优越性[5⇓-7]。但杂交构树原料粗蛋白质含量和缓冲能值高等,导致其在实际生产中青贮成功率较低,营养物质损失较大[8],进而严重阻碍了杂交构树资源的推广和应用。
外源添加乳酸菌制剂是目前国内外学者改善饲料青贮品质的主要手段[9-10]。乳酸菌制剂通过提高青贮原料中乳酸菌数量、改善乳酸菌种类等,进而调节青贮原料微生物区系,确保发酵过程中乳酸菌的主导地位;进而达到防止青贮饲料腐败变质,最大程度保存营养物质的目的[11]。大量研究证实,青贮原料表面附着的乳酸菌数量和种类是影响青贮发酵品质的关键,当青贮原料表面附着乳酸菌数量或种类较少时,需外源添加优质乳酸菌种以确保发酵产品的特性[12]。目前主要采用同质型乳酸菌植物乳杆菌(Lactobacillus plantarum)与异质型乳酸菌布氏乳杆菌(Lactobacillus buchneri)单独或复合接种,并辅以糖源或纤维素酶,以确保杂交构树发酵品质[13⇓-15];但少见杂交构树表面自然附着优良乳酸菌的应用研究。理论上看,发酵原料表面附着乳酸菌相较于外源乳酸菌制剂能够更好地适应植物的生存环境,且在发酵过程中自然附着乳酸菌菌株在定植、存活、生长等方面可能具有一定的优势[16]。鉴于此,本研究以实验室前期杂交构树自然青贮中分离得到的优势乳酸菌——植物乳杆菌、短乳杆菌(Lactobacillus brevis)、屎肠球菌(Enterococcus faecium)为青贮菌剂,通过对营养品质、发酵特性和有氧稳定性的详细分析,明确3种乳酸菌对杂交构树青贮的影响,为今后杂交构树青贮饲料化利用及乳酸菌添加剂的研发提供依据。

1 材料与方法

1.1 试验材料

青贮原料于2021年9月采自新疆维吾尔自治区第八师142团杂交构树种植示范基地(44°52'N、85°50'E,海拔335 m),种植的杂交构树品种为“杂交构树203”,待株高长至1.5 m左右,利用青饲料收获机进行全株收割粉碎,粉碎长度1~2 cm。杂交构树青贮原料营养水平和微生物数量见表1。
表1 杂交构树青贮原料营养水平和微生物数量

Table 1 Nutrient levels and microbial populations of hybrid Broussonetia papyrifera

项目
Item		 干物质
DM/%		 粗蛋
白质
CP/%
DM		 中性洗
涤纤维
NDF/%
DM		 酸性洗
涤纤维
ADF/%
DM		 粗灰分
Ash/
%DM		 可溶性碳
水化合物
WSC/
%DM		 乳酸菌
LAB/
[lg(CFU/
g FM)]		 酵母菌
Yeast/
[lg(CFU/
g FM)]		 霉菌
Mold/
[lg(CFU/
g FM)]		 好氧细菌
AB/
[lg(CFU/
g FM)]
含量
Content		 36.11 20.84 43.80 20.15 10.71 10.54 7.18 6.54 5.30 7.16

供试乳酸菌为前期实验室中分离筛选自杂交构树青贮中的优势乳酸菌,分别为植物乳杆菌(GenBank No. MZ008357)、短乳杆菌(GenBank No. MZ008360)和屎肠球菌(GenBank No. MZ008363)[17]。

1.2 试验设计

杂交构树粉碎后立刻采用真空袋法调制青贮。试验共分7个处理:CK处理仅添加10 mL/kg去离子水,LP处理添加5×106 CFU/g FM植物乳杆菌,LB处理添加5×106 CFU/g FM短乳杆菌,EF处理添加5×106 CFU/g FM屎肠球菌,LP+EF处理复合添加5×106 CFU/g FM植物乳杆菌和屎肠球菌(1∶1),LP+LB处理复合添加5×106 CFU/g FM植物乳杆菌和短乳杆菌(1∶1),LP+LB+EF处理复合添加5×106 CFU/g FM植物乳杆菌、短乳杆菌和屎肠球菌(1∶1∶1)。将各菌种分别在MRS、M17液体培养基中培养,平板计数后确定其数量,按比例配制菌液,以鲜重为基础进行添加。称取500 g样品于带单向排气阀的聚乙烯厌氧袋中分别加入菌液,混合均匀后真空密封(真空袋规格25 cm×40 cm),每个处理5个重复,共计调制35袋。置于18~23 ℃条件下自然发酵60 d后,每个处理随机取3袋保存完好的青贮饲料,混匀后取样,测定营养、发酵指标。另外,采用温度测定仪测定开窖后的温度变化,并在开袋后的第5天测定各处理的pH和主要微生物数量。

1.3 测定指标及方法

营养物质指标主要测定干物质(DM)、粗蛋白质(CP)、粗灰分(Ash)、中性洗涤纤维(NDF)和酸性洗涤纤维(ADF)含量。其中,DM含量采用烘干法(DHG-9620A鼓风干燥机,上海合恒仪器设备有限公司)测定,CP含量采用凯氏定氮法(Kjeltec 8400 FOSS全自动凯氏定氮仪,美国福斯公司)测定,NDF和ADF含量采用范氏洗涤纤维法测定(自制纤维袋,专利号:ZLB01621382211.7和ZLB01621382214.0)[18]。
发酵特征指标主要测定可溶性碳水化合物(WSC)、氨态氮(NH3-N)、4种有机酸[乳酸(LA)、乙酸(AA)、丙酸(PA)、丁酸(BA)]含量及pH。其中,WSC含量采用蒽酮比色法(722可见分光光度计,上海精科实业有限公司)测定,NH3-N含量采用苯酚-次氯酸比色法测定[18-19],有机酸含量采用液相色谱法(Agilent1260高效液相色谱仪,美国安捷伦公司)测定,pH用酸度计(PHSJ3F酸度计,上海雷磁仪器有限公司)测定。
微生物指标主要测定乳酸菌、酵母菌(yeast)、好氧细菌(aerobic bacteria,AB)和霉菌(mold)数量。其中,乳酸菌采用MRS培养基在37 ℃厌氧条件下培养48 h;酵母菌和霉菌分别采用MEA培养基与SCDA培养基在30 ℃厌氧条件下培养24 h;好氧细菌采用PCA培养基在30 ℃好氧条件下培养24 h。上述微生物在培养结束后进行平板计数,并折算微生物数量[20]。
有氧稳定性测定:取400 g青贮样品置于15 cm×20 cm塑料封口袋中用牙签扎数个小孔,套上一个宽松不封口的塑料袋,防止交叉污染和减少水分损失[21],置于阴凉处存放5 d,观测各处理开袋后温度变化,第5天时测定各样品的pH和微生物数量。各指标测定方法同上。

1.4 数据处理与分析

试验数据采用SPSS 20.0统计软件进行单因素方差分析(one-way ANOVA),采用Duncan氏法进行多重比较,以P<0.05作为差异显著性判断标准。

2 结果

2.1 不同乳酸菌处理对杂交构树青贮营养品质的影响

由表2可知,LP和LP+LB+EF处理的DM含量显著高于CK处理(P<0.05)。除LP+EF处理外的其他处理的NDF含量显著低于CK处理(P<0.05)。各处理之间ADF含量均无显著差异(P>0.05)。EF和LP+LB+EF处理的CP含量显著高于除LP处理以外的其他处理(P<0.05)。LP、LB和EF处理的WSC含量显著低于CK处理(P<0.05)。
表2 不同乳酸菌处理对杂交构树青贮营养品质的影响

Table 2 Effects of different lactic acid bacteria treatments on nutritional quality of hybrid Broussonetia papyrifera silage

项目
Items		 处理Treatments SEM P值
P-value
CK LP LB EF LP+
EF		 LP+
LB		 LP+LB+
EF
干物质DM/% 35.01c 36.10ab 35.35bc 35.67bc 35.26bc 35.81bc 36.69a 0.102 0.010
粗蛋白质CP/%DM 17.88bc 18.52ab 17.22c 18.94a 18.04bc 17.50c 19.12a 0.101 <0.001
中性洗涤纤维NDF/%DM 42.78a 38.91b 38.46b 39.39b 40.57ab 39.64b 39.93b 0.303 0.036
酸性洗涤纤维ADF/%DM 20.16 21.04 20.68 20.49 19.43 19.33 18.88 0.635 0.961
可溶性碳水化合物WSC/%DM 7.42a 6.11b 6.30b 6.11b 6.81ab 6.77ab 6.55ab 0.106 0.037

同行数据肩标不同小写字母表示差异显著(P<0.05),相同或无字母表示差异不显著(P>0.05)。下表同。

In the same row, values with different small letter superscripts mean significant difference (P<0.05), while with the same or no letter superscripts mean no significant difference (P>0.05). The same as below.

2.2 不同乳酸菌处理对杂交构树青贮发酵特性及微生物数量的影响

由表3可知,LP和LP+LB+EF处理的pH显著低于CK处理(P<0.05)。CK处理的NH3-N含量显著高于其他处理(P<0.05),且LP处理显著低于EF和LP+EF处理(P<0.05)。各处理的NH3-N/TN均显著低于CK处理(P<0.05)。LP处理的LA含量显著高于EF、LP+LB和CK处理(P<0.05)。EF处理的PA含量显著高于CK处理(P<0.05),而LP、LB和LP+LB处理的PA含量则显著低于CK处理(P<0.05)。LB和LP+LB处理的AA含量显著高于除LP+LB+EF处理外的其他处理(P<0.05)。BA在发酵结束时未检测出。
表3 不同乳酸菌处理对杂交构树青贮发酵特性及微生物数量的影响

Table 3 Effects of different lactic acid bacteria treatments on fermentation characteristics and microbial populations of hybrid Broussonetia papyrifera silage

项目
Items		 处理Treatments SEM P值
P-value
CK LP LB EF LP+
EF		 LP+
LB		 LP+LB+
EF
pH 4.71a 4.42c 4.56abc 4.69a 4.59ab 4.63ab 4.53bc 0.005 0.016
氨态氮NH3-N/%DM 0.65a 0.46c 0.51bc 0.52b 0.53b 0.51bc 0.47bc 0.005 <0.001
氨态氮/总氮NH3-N/TN 0.227a 0.167bc 0.173b 0.173b 0.180b 0.185b 0.157c 0.002 <0.001
乳酸LA/%DM 5.41bc 6.66a 6.25ab 5.09c 5.77abc 5.52bc 6.08ab 0.119 0.046
乙酸AA/%DM 1.77bc 1.89bc 2.13a 1.58c 1.82bc 2.12a 2.07ab 0.043 0.029
丙酸PA/%DM 0.23b 0.14d 0.12d 0.28a 0.25b 0.18c 0.23b 0.000 0.036
丁酸BA/%DM ND ND ND ND ND ND ND
乳酸菌LAB/[lg(CFU/g FM)] 8.52d 8.71b 8.60cd 8.56d 8.77b 8.88a 8.69bc 0.012 <0.001
酵母菌Yeast/[lg(CFU/g FM)] 3.80ab 3.72bc 3.54d 3.88a 3.71bc 3.62cd 3.65cd 0.016 <0.001
霉菌Mold/[lg(CFU/g FM)] 4.15a 3.71b <2.00 <2.00 <2.00 <2.00 <2.00 0.053 0.022
好氧细菌AB/[lg(CFU/g FM)] 7.86a 7.53cd 7.40d 7.45cd 7.41d 7.71b 7.57c 0.016 <0.001

ND:未检测出

not detected。

LP+LB处理的乳酸菌数量显著高于其他处理(P<0.05);除LB和EF处理,其他各处理的乳酸菌数量均显著高于CK处理(P<0.05)。EF处理的酵母菌数量显著高于除CK处理外的其他处理(P<0.05)。除CK和LP处理,其余处理的霉菌数量均<2 lg(CFU/g FM)。CK处理的好氧细菌数量显著高于其他处理(P<0.05)。

2.3 不同乳酸菌处理对杂交构树青贮开袋5 d时pH、微生物数量和有氧稳定性的影响

由表4可知,杂交构树青贮开袋5 d时,仅LP+LB和LP+LB+EF处理的pH与CK处理差异不显著(P>0.05),其他处理的pH均显著高于CK处理(P<0.05)。CK处理的乳酸菌数量均显著低于其他处理(P<0.05),其中LP、EF、LP+EF和LP+LB处理的乳酸菌数量显著高于LB和LP+LB+EF处理(P<0.05)。EF和LP+EF处理的酵母菌数量显著高于CK处理(P<0.05),LB和LP+LB+EF处理的酵母菌数量显著低于CK处理(P<0.05)。CK处理的霉菌和好氧细菌数量均显著高于其他处理(P<0.05),其中LP+LB处理的霉菌数量最低,LP+LB+EF处理的好氧细菌数量最低。LB处理的有氧稳定性显著高于其他处理(P<0.05),且LP+LB和LP+LB+EF处理的有氧稳定性显著高于CK、LP、EF和LP+EF处理。
表4 不同乳酸菌处理对杂交构树青贮开袋5 d时pH、微生物数量与有氧稳定性的影响

Table 4 Effects of different lactic acid bacteria treatments on pH, microbial populations and aerobic stability of hybrid Broussonetia papyrifera silage on the 5th day after opening bags

项目
Items		 处理Treatments SEM P值
P-value
CK LP LB EF LP+
EF		 LP+
LB		 LP+LB+
EF
pH 5.12cd 5.24b 5.20b 5.36a 5.24b 5.15c 5.08d 0.004 <0.001
乳酸菌LAB/[lg(CFU/FM)g] 8.13d 8.64a 8.56b 8.63a 8.65a 8.63a 8.30c 0.007 <0.001
酵母菌Yeast/[lg(CFU/FM)g] 4.52b 4.47b 4.28c 4.87a 4.79a 4.51b 4.24c 0.010 <0.001
霉菌Mold/[lg(CFU/FM)g] 4.26a 4.16b 3.93d 4.07c 4.04c 3.84e 4.01cd 0.004 <0.001
好氧细菌AB/[lg(CFU/FM)g] 9.05a 8.38d 8.40d 8.47c 8.68b 8.64b 8.30e 0.007 <0.001
有氧稳定性Aerobic stability/h 62.80d 59.20d 133.60a 60.40d 58.50d 92.20b 79.30c 28.200 <0.001

3 讨论

3.1 不同乳酸菌处理对杂交构树青贮营养品质的影响

青贮过程中各种微生物和植物本身酶系统的作用使青贮饲料发生一系列生物化学变化,导致营养物质的损失[22]。本研究中,各处理DM和WSC含量在发酵结束时(60 d)均有所下降,其中WSC含量降幅最为明显(降低3.12%~4.43%)。60 d时各处理WSC含量均低于CK处理,其主要由于接种乳酸菌对底物糖源的利用[23];而其中复合菌剂接种(LP+EF、LP+LB和LP+LB+EF处理)相比较单独接种(LP、LB和EF处理)消耗WSC含量少。其原因可能是菌种间的竞争性作用,减少了WSC的消耗。青贮饲料在发酵过程中WSC大部分转化为LA、AA、琥珀酸以及醇类等,其中LA为主要产物,但也有适量AA和少量PA和BA存在[11,22]。目前,对于提高发酵体系LA含量,最为理想的乳酸菌主要为植物乳杆菌(同型发酵乳酸菌)[9]。本研究中接种植物乳杆菌(LP处理)的LA含量相比屎肠球菌(同型发酵乳酸菌,EF处理)和CK处理高,且将2种乳酸菌联合接种时(LP+EF处理),也并未明显抑制LA的生成,其原因可能是乳酸球菌和乳酸杆菌的相互作用所致[24];可能也与乳酸菌利用碳源的种类和效率不同相关[17]。此外,目前大量研究表明,在发酵过程中,当原料WSC含量较低时,微生物更容易产生水解酶系,促使原料纤维类物质的转化,以为自身繁殖提供更多的可利用碳源[25-26]。本研究中,不同乳酸菌处理虽对ADF含量均未产生显著影响,但除2种同型发酵乳酸菌复合接种(LP+EF处理)外,其余单独或复合接种处理均能显著降低发酵体系NDF含量,其原因可能是半纤维素的酸降解作用,进而不需要通过转化纤维类物质来弥补糖源的不足,且仍能确保LA的产生和pH的维持。

3.2 不同乳酸菌处理对杂交构树青贮发酵特性及微生物数量的影响

短乳杆菌通常被认为是严格异型发酵的乳酸菌,但也有报道称其代谢属于兼性异型发酵[27]。本研究中,杂交构树青贮接种短乳杆菌(LB处理)产LA含量与接种植物乳杆菌(LP处理)无显著差异,这与以往推测的短乳杆菌在青贮发酵中的作用规律相同[28];而当二者联合接种(LP+LB处理)时,显著抑制了LA的生成,其主要原因主要为二者对底物营养产生了竞争性抑制[9]。此外,短乳杆菌无论单独或复合接种(LB、LP+LB、LP+LB+EF处理)时均能产生较高的AA含量,其主要原因与异型发酵乳酸菌的特性有关[29],不仅可通过磷酸戊糖途径产生LA和AA;且能以LA为底物产生1,2-丙二醛、乙醇、AA等。
青贮饲料中PA和BA具有刺激性气味,含量较高时严重影响青贮的适口性,因此其含量是衡量青贮饲料优劣的重要指标[30]。本研究中,各处理均含有少量PA,且整个发酵过程中均未检测出BA,其主要原因为发酵底物营养充足,在乳酸菌的主导下形成了较低pH环境,进而有效抑制有害微生物的增殖,减少了有害次级代谢产物的生成[31];同时也可能与构树自身特征(富含具抑菌作用的黄酮类物质)有关[32]。青贮饲料中CP含量的变化与pH高低密切相关,主要由于pH影响牧草中蛋白分解酶的活性,在酸化作用下青贮饲料的蛋白质水解和脱羧基反应会下降,从而抑制蛋白水解酶的活性,降低NH3-N含量[23]。本研究中,青贮发酵60 d后各处理pH在4.4~4.7之间,且各处理NH3-N含量及NH3-N/TN均较低(相比CK处理),说明本研究的的各类乳酸菌菌种组合均能有效形成微酸环境,促使发酵体系中游离氨含量下降,减少NH3-N的产生,有效地降低青贮饲料中蛋白质的降解。
在本研究发酵60 d时,大多数处理均能显著提高发酵体系乳酸菌数量,这与以往研究结果[9-10]基本一致,说明向青贮原料中接种乳酸菌制剂可使乳酸菌在青贮发酵初期迅速成为优势菌群,形成厌氧和微酸环境,使得乳酸菌发酵前期数量增加而后期减少,直至发酵后期数量趋于稳定;而其中单独接种屎肠球菌(EF处理)不能维持发酵体系乳酸菌数量,且显著消耗WSC的同时并未显著提高LA含量,由此说明屎肠球菌产酸量和效率均弱于植物乳杆菌;接种短乳杆菌(LB处理)虽同样不能维持发酵体系乳酸菌数量,但能通过提高AA含量从而促进发酵体系的有氧稳定性。此外,各乳酸菌接种处理均能很好抑制霉菌和好氧细菌的繁殖;其中,对于酵母菌的抑制作用,更多的原因是短乳杆菌的接种(LB、LP+LB、LP+LB+EF处理),进一步证实了短乳杆菌可通过发酵后期代谢产生AA[27],抑制酵母菌的生长。鉴于不同酵母菌在发酵体系中不属于严格的有害或有益菌,且更多的作用机制还尚未明确,在后期需进一步加强研究。

3.3 不同乳酸菌处理对杂交构树青贮开袋5 d时pH、微生物数量和有氧稳定性的影响

青贮饲料的有氧稳定性是评价其价值的重要指标之一。有研究表明,同质型乳酸菌在有氧情况下不能起到增加有氧稳定性的效果[33],这与本研究结果部分一致,本研究中2种同质型乳酸菌单独或联合接种(LP、EF和LP+EF处理)均未改善杂交构树青贮的有氧稳定性,而有短乳杆菌接种(LB、LP+LB和LP+LB+EF处理)时有氧稳定性明显提高,以短乳杆菌单独接种表现最为突出(133.6 h)。一般来说,青贮中的好氧微生物并不全部来自于外界环境,有氧暴露过程为发酵体系潜伏的好氧微生物创造了适宜的繁殖条件[12]。因此,在这一过程中优势乳酸菌数量逐渐减少,酵母菌、霉菌和好氧细菌的数量逐渐增加。以往研究表明,同型发酵乳酸菌产生的能够抑制酵母菌、霉菌等生长繁殖的短链脂肪酸的数量非常少,当青贮饲料与氧气接触后,发酵所产生的LA给乳酸同化型酵母菌提供其生长繁殖所需的底物[34],进而使酵母菌数量迅速增长,这与本研究结果相似,本研究中单独或复合接种屎肠球菌(EF、LP+EF处理)均未明显抑制酵母菌的生长,但当有植物乳杆菌接种(LP、LP+EF处理)时,明显抑制了酵母菌的繁殖。此外,青贮开袋5 d时,各处理的霉菌、好氧细菌数量显著低于CK处理,乳酸菌数量显著高于CK处理,其原因主要为乳酸菌的活性虽不足以维持青贮饲料开窖后长时间的稳定,但短时间内(5 d)仍能通过酸性环境和抗菌物质的产生,抑制好氧微生物生长繁殖[35]。

4 结论

① 复合接种植物乳杆菌、屎肠球菌和短乳杆菌能够最大程度保存DM和CP含量,降低发酵体系NDF含量,减少蛋白质的降解。
② 复合接种植物乳杆菌和短乳杆菌能够维持发酵和青贮开袋5 d内的乳酸菌数量,进而通过提高AA的生成抑制霉菌、好氧细菌的生长,提高杂交构树青贮的有氧稳定性。

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王旭哲, 张凡凡, 马春晖, 等. 同/异型乳酸菌对青贮玉米开窖后品质及微生物的影响[J]. 农业工程学报, 2018, 34(10):296-304.

WANG X Z, ZHANG F F, MA C H, et al. Corn silage fermentation quality and microbial populations as influenced by adding homo-and hetero-fermentative bacteria after silos opened[J]. Transactions of the Chinese Society of Agricultural Engineering, 2018, 34(10):296-304. (in Chinese)

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