Chinese Journal of Animal Nutrition >

2025 , Vol. 37 >Issue 9: 5923 - 5935

DOI: https://doi.org/10.12418/CJAN2025.482

RESEARCH PAPER

Effects of Streptococcus alactolyticus from Chickens on Growth Performance, Meat Quality, Antioxidant Capacity and Expression of Inflammatory Factors in Muscle of White-Feathered Broiler Chickens

  • ZHOU Qianxi , 1, 2, 3, 4, 5 ,
  • XIE Kun 1, 2, 3, 4, 5 ,
  • ZHANG Haihan 1, 2, 3, 4, 5 ,
  • SONG Zehe 1, 2, 3, 4, 5 ,
  • HE Xi , 1, 2, 3, 4, 5, *
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  • 1 College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China
  • 2 Engineering Research Center for Feed Safety and Efficient Utilization, Ministry of Education, Changsha 410128, China
  • 3 Hunan Poultry Safety Production Engineering Technology Research Center, Changsha 410128, China
  • 4 Provincial and Ministerial Co-Built Collaborative Innovation Center for the Production of High-Quality Livestock and Poultry Products, Changsha 410128, China
  • 5 Yuelu Mountain Laboratory, Changsha 410128, China
* professor, E-mail:

Received date: 2025-01-14

  Online published: 2025-09-12

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Abstract

This experiment aimed to investigate the effects of dietary Streptococcus alactolyticus from chickens on growth performance, meat quality, antioxidant capacity and expression of inflammatory factors in muscle of white-feathered broiler chickens. The Streptococcus alactolyticus FM231008 used in the experiment was screened out from broiler chicken feces by our laboratory, and was identified as Streptococcus alactolyticus with the sequence compared with the NCBI database. A total of 360 one-day-old healthy Arbor Acres (AA) white-feathered broiler chickens were selected and randomly divided into 5 groups, with 6 replicates in each group and 12 chickens in each replicate. The chickens in the control group (CON group) were fed a basal diet, while those in the experimental groups were fed the basal diets supplemented with 2.5×1010 (T1 group), 5.0×1010 (T2 group), 7.5×1010 (T3 group) and 10.0×1010 CFU/kg (T4 group) Streptococcus alactolyticus powder, respectively. The experiment lasted for 42 days. The results showed as follows: 1) there were no significant differences in growth performance and slaughter performance among all groups (P>0.05). 2) Compared with CON group, the pH45 min in breast muscle in T3 group was significantly decreased (P<0.05), the pH24 h and water loss rate in breast muscle in T3 and T4 groups were significantly decreased (P<0.05), the redness (a*) and yellowness (b*) values in breast muscle in T1 and T2 groups were significantly decreased (P<0.05), and the shear force in breast muscle in each experimental group was significantly decreased (P<0.05). 3) At 21 days of age, compared with CON group, the activities of total superoxide dismutase (T-SOD) and catalase (CAT) in serum in T1 and T4 groups were significantly increased (P<0.05), and the malondialdehyde (MDA) content in serum was significantly decreased (P<0.05); the glutathione peroxidase (GSH-Px) activity in serum in T1 group was significantly increased (P<0.05); the CAT activity in serum in T2 group was significantly increased (P<0.05); the MDA content in serum in T3 group was significantly decreased (P<0.05). Compared with CON group, the MDA content in breast muscle in each experimental group was significantly decreased (P<0.05), and the total antioxidant capacity (T-AOC) in breast muscle in T2 and T4 groups was significantly increased (P<0.05). At 42 days of age, compared with CON group, the CAT activity in serum in each experimental group was significantly increased (P<0.05); the T-AOC in serum in T1 and T4 groups was significantly increased (P<0.05); the GSH-Px activity in serum in T2, T3 and T4 groups was significantly increased (P<0.05), and the MDA content in serum was significantly decreased (P<0.05); the T-SOD activity in serum in T3 group was significantly increased (P<0.05). Compared with CON group, the MDA content in breast muscle in T1, T3 and T4 groups was significantly decreased (P<0.05), and the T-AOC in breast muscle in T2 group was significantly increased (P<0.05). 4) At 21 days of age, compared with CON group, the mRNA relative expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in breast muscle in each experimental group were significantly decreased (P<0.05), the interleukin-2 (IL-2) mRNA relative expression level in breast muscle in T1, T3 and T4 groups was significantly decreased (P<0.05), and the interleukin-10 (IL-10) mRNA relative expression level in breast muscle in T1 and T3 groups was significantly decreased (P<0.05). At 42 days of age, compared with CON group, the mRNA relative expression levels of IL-1β, IL-6 and TNF-α in breast muscle in each experimental group were significantly decreased (P<0.05), the IL-2 mRNA relative expression level in breast muscle in T2 group was significantly decreased (P<0.05), and the IL-10 mRNA relative expression level in breast muscle in T2 and T3 groups was significantly decreased (P<0.05). In conclusion, under the conditions of this experiment, appropriate dose of dietary Streptococcus alactolyticus from chickens can improve the meat quality and alleviate oxidative stress by enhancing antioxidant capacity and regulating inflammatory responses of white-feathered broiler chickens.

Key words： Streptococcus alactolyticus from chickens; white-feathered broiler chickens; growth performance; meat quality; antioxidant capacity; inflammatory factors

Cite this article

ZHOU Qianxi , XIE Kun , ZHANG Haihan , SONG Zehe , HE Xi . Effects of Streptococcus alactolyticus from Chickens on Growth Performance, Meat Quality, Antioxidant Capacity and Expression of Inflammatory Factors in Muscle of White-Feathered Broiler Chickens[J]. Chinese Journal of Animal Nutrition, 2025 , 37(9) : 5923 -5935 . DOI: 10.12418/CJAN2025.482

鸡肉因其高蛋白质、低脂肪和低热量的特点深受消费者喜爱,也是目前仅次于猪肉的第二大肉类产品,具有极高的发展潜力[1]。白羽肉鸡因其饲养周期短、饲料回报率高和肉产量高等特点而被大量饲养,成为我国禽肉消费之首[2]。根据中国畜牧业协会数据显示,2023年我国白羽肉鸡出栏量为71.95亿只,同比增长18.14%,白羽肉鸡出栏量占我国肉鸡出栏量的55.25%。但是近年来,白羽肉鸡木质化鸡胸肉、白条纹肉等劣质肉问题不断发生,严重影响白羽肉鸡鸡肉产品质量[3-4]。
非解乳糖链球菌(Streptococcus alactolyticus)为革兰氏阳性菌,呈链状排列,无芽孢,无鞭毛且无运动性[5]。非解乳糖链球菌最早是从猪和鸡的肠道中分离出来,且发现该菌可能与肉鸡的生长繁殖与肠道健康以及肉质改善有着密切的关系。研究发现,其能够提升肠道谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)等抗氧化酶活性,而这些抗氧化酶可能通过循环系统影响肌肉组织,减少肌肉脂质过氧化和肌纤维异常,从而改善肉质[6-7]。此外,非解乳糖链球菌还表现出极强的耐酸和耐胆盐特性,同时此菌株基因组中包含151个与生物黏附相关的基因,可通过表面蛋白[如热不稳定延伸因子(elongation factor thermo unstable,EF-Tu)]介导与宿主肠上皮的黏附,因此此菌种具备作为饲用益生菌开发的巨大潜力[5]。本团队前期研究同样发现,肉鸡肠道非解乳糖链球菌相对丰度与木质化程度呈显著负相关关系[8],并进一步从鸡盲肠中筛选到1株具有较好耐酸、耐胆盐活性的非解乳糖链球菌FM231008。因此,本试验旨在研究饲粮中添加非解乳糖链球菌菌粉对肉鸡生长性能、肉品质、抗氧化能力和肌肉炎性因子表达的影响,为进一步探索应对肉鸡肉质问题的措施提供参考。

1 材料和方法

1.1 试验菌株

鸡源细菌非解乳糖链球菌FM231008是本实验室从肉鸡粪便中筛出,并经过菌株纯化培养后,筛选出的一株厌氧菌菌株。经TSINGKE细菌基因组DNA提取试剂盒测序(扩增及测序引物信息见表1)送样并返回序列结果,并经NCBI数据库比对,该菌株与模式菌株非解乳糖链球菌ATCC 43077相似性达100%,比对序列号为NR_041781。利用MEGA-X软件,进行ClustalW多重序列对比,并采用邻接法构建系统发育树,对其进行系统发育分析如图1所示。从系统发育树中可以看到,这些细菌由一个属组成,其中与菌株FM231008的16S rDNA序列比对相似度最高的菌株几乎全部属于链球菌属(Streptococcus)。根据细菌测定的16S rDNA碱基序列,比对模式菌株,对所鉴定的细菌确定种属及命名,命名规则采用基于16S层面的中国海洋微生物菌种保藏管理中心(Marine Culture Collection of China,MCCC)原则,该菌株暂定名为非解乳糖链球菌FM231008。
表1 非解乳糖链球菌FM231008扩增及测序引物信息

Table 1 Amplification and sequencing primer information for Streptococcus alactolyticus FM231008

项目
Items		 名称
Names		 序列
Sequences
(5'—3')		 扩增序列
Ampliconic
sequence		 PCR长度
PCR length/bp
扩增引物
Amplification primer		 27F
1492R		 AGTTTGATCMTGGCTCAG
GGTTACCTTGTTACGACTT		 16S rDNA 1 500左右
测序引物
Sequencing primer		 789F
932R		 CGAAAGCGTGGGGAGCAAAC
CCCCGTCAATTCCTTTGAGT		 16S rDNA 1 500左右
图1 非解乳糖链球菌FM231008系统发育树

Streptococcus pasteurianus:巴氏链球菌;Streptococcus equinus:马链球菌;Streptococcus alactolyticus:非解乳糖链球菌;Streptococcus macedonicus:马其顿链球菌;Streptococcus sanguinis:血链球菌;Streptococcus danieliae:丹尼尔链球菌。

Fig.1 Phylogenetic tree of Streptococcus alactolyticus FM231008

1.2 试验设计和饲粮

本试验各项操作均经过湖南农业大学动物伦理委员会的审查和批准(批准编号:HAU ACC 2023DK JQ016)。试验选取1日龄健康白羽肉鸡360只,随机分为5组,每组6个重复,每个重复12只鸡。对照组(CON组)饲喂基础饲粮,试验组在基础饲粮中分别添加2.5×1010(T1组)、5.0×1010(T2组)、7.5×1010(T3组)和10.0×1010 CFU/kg(T4组)非解乳糖链球菌菌粉。鸡源非解乳糖链球菌菌粉由本团队实验室制备冻干所得,菌粉浓度为1×1013 CFU/mL。试验期42 d。
基础饲粮参照Aviagen公司爱拔益加(AA)肉鸡品种推荐标准(2014年)配制,分前期(1~21日龄)和后期(22~42日龄)2个阶段,其组成及营养水平见表2。饲粮代谢能参考《中国饲料成分及营养价值表(2020年第31版)》计算。
表2 基础饲料组成及营养水平(风干基础)

Table 2 Composition and nutrient levels of basal diets (air-dry basis) %

原料
Ingredients		 1~21日龄
1 to 21
days of age		 22~42日龄
22 to 42
days of age		 营养水平
Nutrient levels2)		 1~21日龄
1 to 21
days of age		 22~42日龄
22 to 42
days of age
玉米Corn 56.51 62.21 代谢能ME/(MJ/kg) 12.71 13.01
豆粕Soybean meal (43% CP) 35.30 29.20 粗蛋白质CP 20.65 18.28
石粉Limestone 1.52 1.60 赖氨酸Lys 1.27 1.09
大豆油Soybean oil 4.50 4.90 蛋氨酸Met 0.47 0.41
磷酸氢钙CaHPO4 1.00 1.00 蛋氨酸+胱氨酸Met+Cys 0.95 0.86
赖氨酸盐酸盐Lys·HCl (98%) 0.35 0.30 苏氨酸Thr 0.83 0.74
DL-蛋氨酸DL-Met (98%) 0.16 0.13 色氨酸Trp 0.24 0.20
L-苏氨酸L-Thr (98%) 0.06 0.06 钙Ca 0.90 0.91
食盐NaCl 0.30 0.30 总磷TP 0.54 0.52
预混料Premix1) 0.30 0.30
合计Total 100.00 100.00

1)预混料为每千克饲粮提供 The premix provided the following per kg of diets:VA 12 000 IU,VD3 2 500 IU,VE 20.0 mg,VK3 3.0 mg,VB1 3.0 mg,VB2 8.0 mg,VB6 7.0 mg,VB12 0.03 mg,泛酸 pantothenic acid 20.0 mg,烟酸 nicotinic acid 50.0 mg,生物素 biotin 0.1 mg,叶酸 folic acid 1.5 mg,Fe 80 mg,Cu 17.5 mg,I 1.5 mg,Zn 105 mg,Mn 124 mg。

2)代谢能为计算值,其余均为实测值。ME was a calculated value, while the others were measured values.

1.3 饲养管理

试验于湖南省长沙市长沙县开慧镇锡福小院进行。试验开始前,对鸡舍进行彻底清扫,并使用“甲醛+高锰酸钾”进行熏蒸消毒,3 d后进行通风。试验采用全封闭式笼养,以上、中和下3层笼养方式饲养。整个试验阶段鸡只自由采食和饮水,饲养管理参考《AA肉仔鸡饲养管理手册》进行。

1.4 测定指标及方法

1.4.1 饲粮营养成分

饲粮粗蛋白质含量参照GB/T 6432—2018方法测定,钙含量参照GB/T 6436—2018方法测定,总磷含量参照GB/T 6437—2018方法测定,氨基酸含量参照GB/T 18246—2019方法测定。

1.4.2 生长性能

分别于21和42日龄以重复为单位对试验鸡进行称重,试验期间记录试验鸡采食量和死淘情况,计算平均日增重(ADG)、平均日采食量(ADFI)和料重比(F/G)。

1.4.3 屠宰性能

于试验第42天,每个重复挑选1只体重接近平均体重的试验鸡空腹称重后颈动脉放血处死,依次称量半净膛、全净膛、胸肌、腿肌以及腹脂的重量并计算各个屠宰性能指标,具体测定方法参照《家禽生产性能名词术语和度量统计方法》(NY/T 823—2020)进行。屠宰性能指标计算公式如下:
屠宰率(%)=(屠体重/活重)×100;
半净膛率(%)=(半净膛重/活重)×100;
全净膛率(%)=(全净膛重/活重)×100;
胸肌率(%)=(胸肌重/全净膛重)×100;
腿肌率(%)=(腿肌重/全净膛重)×100;
腹脂率(%)=(腹脂重/全净膛重)×100。

1.4.4 肉品质

胸肌剪切力参照《肉嫩度的测定 剪切力测定法》(NY/T 1180—2006)方法测定,亮度(L*)、红度(a*)和黄度(b*)值以及失水率参照《畜禽肉质的测定》(NY/T 1333—2007)方法测定,屠宰后45 min和24 h时pH采用pH计(Opto-Star,德国)测定。

1.4.5 抗氧化能力

分别于21和42日龄,每个重复挑选1只体重接近平均体重的试验鸡进行颈静脉取血,血样收集于10 mL抗凝管中(收集量约为管容积的2/3),倾斜放置15 min后,1 006×g离心10 min,收集上层血清,-80 ℃冰箱保存,待测。屠宰后取肉鸡胸肌样品,置于液氮中,于-80 ℃冰箱保存,待测。按照试剂盒(南京建成生物工程研究所)说明书步骤测定血清和胸肌总抗氧化能力(T-AOC)、丙二醛(MDA)含量以及总超氧化物歧化酶(T-SOD)、GSH-Px和过氧化氢酶(CAT)活性。

1.4.6 肌肉炎性因子表达

采用TRIzol试剂盒(南京诺唯赞生物科技股份有限公司)提取肉鸡胸肌总RNA,并采用1.0%浓度的琼脂糖凝胶电泳检测RNA的完整性。采用Evo M-MLV反转录预混型试剂盒Ver.2(含去除gDNA试剂,湖南艾科瑞生物工程有限公司)将RNA反转录成cDNA。利用NCBI中相应基因组序列信息,通过NCBI中的Pick Primers进行引物设计,引物序列见表3。采用荧光定量PCR仪[LightCycler 480 Ⅱ,罗氏诊断产品(上海)有限公司],根据SYBR Green Pro Taq HS预混型qPCR试剂盒(产品货号AG11701,湖南艾科瑞生物工程有限公司)说明书进行实时荧光定量PCR反应。以β-肌动蛋白(β-actin)作为内参基因,采用2-△△Ct法计算目的基因mRNA相对表达量。
表3 引物序列

Table 3 Primer sequences

基因
Genes		 登录号
Accession number		 引物序列
Primers sequences(5'—3')		 产物长度
Product length/bp
β-肌动蛋白β-actin NM 205518.2 F:CACCACAGCCGAGAGAGAAAT
R:TGACCATCAGGGAGTTCATAGC		 135
白细胞介素-1β IL-1β NM_204524.2 F:ACTGGGCATCAAGGGCTACA
R:GCTGTCCAGGCGGTAGAAGA		 142
白细胞介素-2 IL-2 NC_052535.1 F:CTCCGAGTTCTCCCCGAAAG
R:CAAAGTTGGTCAGTTCATGGAGA		 190
白细胞介素-6 IL-6 NM_204628.2 F:AAATCCCTCCTCGCCAATCT
R:CCCTCACGGTCTTCTCCATAAA		 132
白细胞介素-10 IL-10 NM_001004414.4 F:GGGTGAAGTTTGAGGAAATTAAGGA
R:TCATCTGTGTAGAAGCGCAGCA		 218
肿瘤坏死因子-α TNF-α XM_046927265.1 F:GCCCTTCCTGTAACCAGATG
R:ACACGACAGCCAAGTCAACG		 158

1.5 数据统计分析

试验数据经Excel 2016初步整理后,采用SPSS 26.0软件进行单因素方差分析(one-way ANOVA),并采用Duncan氏法进行多重比较,同时进行线性和二次效应分析。结果数据采用“平均值±标准差”表示,P<0.05表示差异显著。

2 结果与分析

2.1 非解乳糖链球菌对白羽肉鸡生长性能的影响

由表4可知,试验期间各组间白羽肉鸡各阶段生长性能指标均无显著差异(P>0.05),表明饲粮中添加非解乳糖链球菌对白羽肉鸡生长性能无显著影响。
表4 非解乳糖链球菌对白羽肉鸡生长性能的影响

Table 4 Effects of Streptococcus alactolyticus on growth performance of white-feathered broiler chickens

项目
Items		 组别Groups P值P-value
CON T1 T2 T3 T4 处理
Treatment		 线性
Linear		 二次
Quadratic
1~21日龄1 to 21 days of age
初重IBW/g 46.3±3.3 44.6±0.5 46.4±2.1 45.5±0.7 45.7±0.6 0.206 0.312 0.068
末重FBW/g 656.7±14.2 645.6±17.0 654.4±11.5 655.8±13.3 663.6±22.7 0.462 0.207 0.892
平均日增重ADG/(g/d) 29.1±0.6 28.6±0.8 28.9±0.5 29.1±0.6 29.4±1.1 0.497 0.392 0.323
平均日采食量
ADFI/(g/d)		 42.1±0.9 41.4±1.2 42.3±0.7 42.6±1.0 43.7±1.0 0.315 0.286 0.116
22~42日龄22 to 42 days of age
末重FBW/g 2 227.8±37.8 2 222.7±96.1 2 186.5±84.5 2 193.5±54.1 2 241.6±76.7 0.214 0.334 0.157
平均日增重ADG/(g/d) 74.8±2.0 75.1±3.9 73.0±2.8 73.3±1.6 75.2±3.5 0.881 0.735 0.674
平均日采食量
ADFI/(g/d)		 111.2±4.6 112.5±7.6 112.3±5.8 111.4±3.8 113.3±6.9 0.939 0.673 0.893
1~42日龄1 to 42 days of age
平均日增重ADG/(g/d) 52.0±0.9 51.9±2.3 51.0±2.0 51.2±1.3 52.3±1.8 0.653 0.768 0.465
平均日采食量
ADFI/(g/d)		 76.6±1.8 76.9±2.1 77.3±1.3 77.3±2.3 78.5±2.1 0.881 0.784 0.673
料重比F/G 1.48±0.03 1.48±0.01 1.51±0.02 1.51±0.04 1.50±0.04 0.239 0.136 0.175

同行数据肩标相同字母或无字母表示差异不显著(P>0.05),不同字母表示差异显著(P<0.05)。下表同。

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

2.2 非解乳糖链球菌对白羽肉鸡屠宰性能的影响

由表5可知,各组间白羽肉鸡屠宰性能指标均无显著差异(P>0.05),表明饲粮中添加非解乳糖链球菌对白羽肉鸡屠宰性能无显著影响。随着饲粮中非解乳糖链球菌添加剂量的提高,白羽肉鸡屠宰率呈现先升高后降低的二次变化趋势(P<0.05)。
表5 非解乳糖链球菌对白羽肉鸡屠宰性能的影响

Table 5 Effects of Streptococcus alactolyticus on slaughter performance of white-feathered broiler chickens %

项目
Items		 组别Groups P值P-value
CON T1 T2 T3 T4 处理
Treatment		 线性
Linear		 二次
Quadratic
屠宰率
Dressing percentage		 92.7±1.3 92.1±1.1 92.7±1.2 92.6±1.5 92.5±1.5 0.789 0.893 0.045
半净膛率
Semi-eviscerated rate		 83.4±1.9 83.6±2.3 84.1±2.3 83.2±2.6 83.7±1.5 0.905 0.826 0.145
全净膛率Eviscerated rate 71.1±1.2 72.2±2.6 71.0±2.4 70.4±1.8 70.8±1.7 0.254 0.532 0.407
胸肌率Breast muscle rate 26.7±0.9 27.5±1.7 26.8±2.0 26.8±1.5 27.6±1.4 0.595 0.473 0.326
腿肌率Leg muscle rate 21.1±1.1 21.3±1.4 21.7±1.3 21.2±1.2 21.1±1.2 0.515 0.414 0.302
腹脂率Abdominal fat rate 1.7±0.2 1.6±0.2 1.6±0.4 1.7±0.5 1.7±0.2 0.945 0.992 0.254

2.3 非解乳糖链球菌对白羽肉鸡肉品质的影响

由表6可知,与CON组相比,T3组白羽肉鸡胸肌pH45 min显著降低(P<0.05),T3组和T4组胸肌pH24 h和失水率显著降低(P<0.05),T1组和T2组胸肌a*和b*值显著降低(P<0.05),各试验组胸肌剪切力均显著降低(P<0.05);各组间胸肌L*值无显著差异(P>0.05)。随着饲粮中非解乳糖链球菌添加剂量的提高,白羽肉鸡胸肌失水率和b*值呈线性和二次变化(P<0.05),胸肌pH24 h呈二次变化(P<0.05),胸肌剪切力呈线性提高(P<0.05)。
表6 非解乳糖链球菌对白羽肉鸡肉品质的影响

Table 6 Effects of Streptococcus alactolyticus on meat quality of white-feathered broiler chickens

项目
Items		 组别Groups P值P-value
CON T1 T2 T3 T4 处理
Treatment		 线性
Linear		 二次
Quadratic
pH45 min 6.67±0.18a 6.61±0.18ab 6.66±0.17a 6.49±0.20b 6.59±0.22ab 0.022 0.109 0.326
pH24 h 5.93±0.08a 5.89±0.08ab 5.88±0.14ab 5.83±0.07bc 5.80±0.10c 0.005 0.056 0.002
失水率Water loss rate/% 8.56±0.22a 6.24±0.14b 8.08±0.26ab 6.41±0.23b 6.66±0.17b 0.022 0.014 0.005
亮度L* 48.94±1.93 49.79±1.45 50.50±1.87 49.01±1.89 49.83±1.87 0.193 0.254 0.145
红度a* 5.88±0.89a 5.11±0.51c 5.26±0.58bc 5.67±0.73ab 5.52±0.57abc 0.025 0.606 0.407
黄度b* 9.36±0.94a 8.35±0.53b 7.24±0.98c 8.56±0.92ab 9.14±0.50ab 0.001 0.005 0.006
剪切力Shear force/N 12.43±0.88a 9.89±1.23b 11.06±0.80b 9.99±1.28b 11.17±0.68b 0.001 0.044 0.050

2.4 非解乳糖链球菌对白羽肉鸡血清抗氧化指标的影响

由表7可知,21日龄,与CON组相比,T1组和T4组白羽肉鸡血清T-SOD和CAT活性显著提高(P<0.05),血清MDA含量显著降低(P<0.05);T1组血清GSH-Px活性显著提高(P<0.05);T2组血清CAT活性显著提高(P<0.05);T3组血清MDA含量显著降低(P<0.05)。随着饲粮中非解乳糖链球菌添加剂量的提高,白羽肉鸡血清CAT活性和MDA含量呈线性变化(P<0.05)。
表7 非解乳糖链球菌对白羽肉鸡血清抗氧化指标的影响

Table 7 Effects of Streptococcus alactolyticus on serum antioxidant indices of white-feathered broiler chickens

项目
Items		 组别Groups P值P-value
CON T1 T2 T3 T4 处理
Treatment		 线性
Linear		 二次
Quadratic
21日龄21 days of age
总抗氧化能力
T-AOC/(U/mL)		 23.74
±1.85		 23.43
±1.01		 24.98
±1.18		 23.74
±1.18		 25.90
±1.01		 0.074 0.459 0.116
总超氧化物歧化酶
T-SOD/(U/mL)		 213.41
±11.12c		 236.37
±19.36ab		 214.06
±24.91bc		 227.61
±16.28abc		 253.65
±17.05a		 0.014 0.900 0.701
过氧化氢酶
CAT/(U/mL)		 3.21
±0.24c		 3.74
±0.16ab		 3.89
±0.32a		 3.45
±0.24bc		 3.66
±0.12ab		 0.015 0.006 0.285
谷胱甘肽过氧化物酶
GSH-Px/(U/mL)		 1 065.60
±27.43b		 1 221.60
±34.28a		 1 032.80
±97.62b		 1 070.40
±140.71b		 1 150.80
±82.62ab		 0.048 0.180 0.147
丙二醛
MDA/(nmol/mL)		 3.86
±0.47a		 3.02
±0.41b		 3.82
±0.22a		 2.94
±0.29b		 2.94
±0.08b		 0.001 0.006 0.922
42日龄42 days of age
总抗氧化能力
T-AOC/(U/mL)		 18.19
±0.61c		 20.35
±0.71ab		 19.12
±0.71bc		 19.12
±1.59bc		 21.58
±1.59a		 0.007 0.320 0.398
总超氧化物歧化酶
T-SOD/(U/mL)		 252.55
±14.98b		 259.27
±31.34b		 280.42
±24.51b		 321.58
±40.19a		 257.65
±7.08b		 0.003 0.002 0.031
过氧化氢酶
CAT/(U/mL)		 3.22
±0.08c		 3.52
±0.08b		 3.47
±0.14b		 4.03
±0.12a		 3.46
±0.15b		 0.001 0.001 0.003
谷胱甘肽过氧化物酶
GSH-Px/(U/mL)		 1 044.20
±37.51c		 1 067.21
±33.75bc		 1 112.81
±35.32ab		 1 116.83
±42.31ab		 1 140.11
±37.51a		 0.029 0.794 0.004
丙二醛
MDA/(nmol/mL)		 4.56
±0.29a		 4.35
±0.43ab		 3.77
±0.54bc		 3.63
±0.15c		 3.81
±0.41bc		 0.016 0.472 0.001
42 日龄,与CON组相比,各试验组白羽肉鸡血清CAT活性均显著提高(P<0.05);T1组和T4组血清T-AOC显著提高(P<0.05);T2组、T3组和T4组血清GSH-Px活性显著提高(P<0.05),血清MDA含量显著降低(P<0.05);T3组血清T-SOD活性显著提高(P<0.05)。随着饲粮中非解乳糖链球菌添加剂量的提高,白羽肉鸡血清T-SOD和CAT活性呈线性和二次变化(P<0.05),血清GSH-Px活性和MDA含量呈二次变化(P<0.05)。

2.5 非解乳糖链球菌对白羽肉鸡肌肉抗氧化指标的影响

由表8可知,21日龄,与CON组相比,各试验组白羽肉鸡胸肌MDA含量显著降低(P<0.05),T2组和T4组胸肌T-AOC显著提高(P<0.05)。随着饲粮中非解乳糖链球菌添加剂量的提高,白羽肉鸡胸肌T-AOC和CAT活性呈线性和二次变化(P<0.05),胸肌T-SOD和GSH-Px活性以及MDA含量呈线性变化(P<0.05)。
表8 非解乳糖链球菌对白羽肉鸡肌肉抗氧化指标的影响

Table 8 Effects of Streptococcus alactolyticus on antioxidant indices in muscle of white-feathered broiler chickens

项目
Items		 组别Groups P值P-value
CON T1 T2 T3 T4 处理
Treatment		 线性
Linear		 二次
Quadratic
21日龄21 days of age
总抗氧化能力
T-AOC/(U/mg prot)		 2.22
±0.10b		 2.43
±0.10ab		 2.61
±0.10a		 2.39
±0.10ab		 2.52
±0.10a		 0.015 0.036 0.035
总超氧化物歧化酶
T-SOD/(U/mg prot)		 165.06
±0.90a		 156.73
±3.30b		 161.24
±2.90ab		 158.00
±2.40b		 164.43
±0.90a		 0.005 0.001 0.988
过氧化氢酶
CAT/(U/mg prot)		 1.54
±0.02a		 1.45
±0.09abc		 1.41
±0.11bc		 1.36
±0.01c		 1.50
±0.03ab		 0.020 0.025 0.017
谷胱甘肽过氧化物酶
GSH-Px/(U/mg prot)		 44.95
±1.10a		 38.14
±1.70c		 42.13
±2.40ab		 41.93
±0.80ab		 41.11
±2.30bc		 0.015 0.004 0.383
丙二醛
MDA/(nmol/mg prot)		 4.37
±0.10a		 2.96
±0.10b		 3.50
±0.20b		 3.38
±0.30b		 3.47
±0.30b		 0.001 0.001 0.171
42日龄42 days of age
总抗氧化能力
T-AOC/(U/mg prot)		 2.46
±0.20b		 2.69
±0.30ab		 3.19
±0.20a		 2.53
±0.30b		 2.71
±0.30ab		 0.048 0.085 0.806
总超氧化物歧化酶
T-SOD/(U/mg prot)		 168.94
±0.90a		 150.76
±3.00c		 163.28
±1.20ab		 158.66
±3.90b		 158.98
±3.30b		 0.002 0.001 0.986
过氧化氢酶
CAT/(U/mg prot)		 1.83
±0.10		 1.63
±0.30		 1.46
±0.20		 1.35
±0.30		 1.52
±0.20		 0.171 0.310 0.025
谷胱甘肽过氧化物酶
GSH-Px/(U/mg prot)		 84.97
±2.70		 73.98
±3.60		 80.03
±2.80		 75.62
±3.30		 81.55
±2.40		 0.089 0.019 0.638
丙二醛
MDA/(nmol/mg prot)		 4.06
±0.20a		 3.02
±0.40b		 3.76
±0.20a		 2.97
±0.30b		 2.61
±0.20b		 0.001 0.190 0.033
42日龄,与CON组相比,T1组、T3组和T4组白羽肉鸡胸肌MDA含量显著降低(P<0.05),T2组胸肌T-AOC显著提高(P<0.05);各组间胸肌CAT和GSH-Px活性无显著差异(P>0.05)。随着饲粮中非解乳糖链球菌添加剂量的提高,白羽肉鸡胸肌T-SOD和GSH-Px活性呈线性提高(P<0.05),胸肌CAT活性和MDA含量呈二次变化(P<0.05)。

2.6 非解乳糖链球菌对白羽肉鸡肌肉炎性因子表达的影响

由表9可知,21日龄,与CON组相比,各试验组白羽肉鸡胸肌白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)mRNA相对表达量显著降低(P<0.05),T1组、T3组和T4组胸肌白细胞介素-2(IL-2)mRNA相对表达量显著降低(P<0.05),T1组和T3组胸肌白细胞介素-10(IL-10)mRNA相对表达量显著降低(P<0.05)。随着饲粮中非解乳糖链球菌添加剂量的提高,白羽肉鸡胸肌IL-1β、IL-6和TNF-α mRNA相对表达量呈线性和二次变化(P<0.05),胸肌IL-2和IL-10 mRNA相对表达量呈线性变化(P<0.05)。
表9 非解乳糖链球菌对白羽肉鸡肌肉炎性因子表达的影响

Table 9 Effects of Streptococcus alactolyticus on expression of inflammatory factors in muscle of white-feathered broiler chickens

项目
Items		 组别Groups P值P-value
CON T1 T2 T3 T4 处理
Treatment		 线性
Linear		 二次
Quadratic
21日龄21 days of age
白细胞介素-1β IL-1β 1.00±0.08a 0.37±0.05c 0.76±0.06b 0.39±0.05c 0.22±0.03d 0.001 0.001 0.001
白细胞介素-2 IL-2 1.00±0.06a 0.75±0.09b 0.87±0.06ab 0.84±0.08b 0.80±0.09b 0.009 0.005 0.443
白细胞介素-6 IL-6 1.00±0.07a 0.85±0.08b 0.49±0.09c 0.30±0.02d 0.46±0.04c 0.001 0.004 0.001
白细胞介素-10 IL-10 1.00±0.08a 0.55±0.11c 0.98±0.09a 0.78±0.12b 0.93±0.11ab 0.001 0.001 0.157
肿瘤坏死因子-α TNF-α 1.00±0.09a 0.52±0.08b 0.61±0.09b 0.41±0.09c 0.52±0.01b 0.001 0.001 0.001
42日龄42 days of age
白细胞介素-1β IL-1β 1.00±0.09a 0.54±0.07c 0.12±0.02d 0.13±0.03d 0.82±0.10b 0.001 0.001 0.001
白细胞介素-2 IL-2 1.00±0.10a 0.99±0.13a 0.79±0.06b 0.96±0.09a 0.85±0.07ab 0.018 0.654 0.029
白细胞介素-6 IL-6 1.00±0.01a 0.24±0.03cd 0.18±0.01d 0.30±0.01c 0.63±0.09b 0.001 0.001 0.001
白细胞介素-10 IL-10 1.00±0.08a 0.96±0.12ab 0.56±0.09c 0.78±0.08b 0.91±0.09ab 0.001 0.048 0.002
肿瘤坏死因子-α TNF-α 1.00±0.08a 0.18±0.04b 0.13±0.03b 0.25±0.05b 0.22±0.01b 0.001 0.001 0.001
42日龄,与CON组相比,各试验组白羽肉鸡胸肌IL-1β、IL-6和TNF-α mRNA相对表达量显著降低(P<0.05),T2组胸肌IL-2 mRNA相对表达量显著降低(P<0.05),T2组和T3组胸肌IL-10 mRNA相对表达量显著降低(P<0.05)。随着饲粮中非解乳糖链球菌添加剂量的提高,白羽肉鸡胸肌IL-1β、IL-6、IL-10和TNF-α mRNA相对表达量呈线性和二次变化(P<0.05),胸肌IL-2 mRNA相对表达量呈二次变化(P<0.05)。

3 讨论

3.1 非解乳糖链球菌对白羽肉鸡生长性能的影响

非解乳糖链球菌作为一种肠道益生菌,通过饲粮摄入后进入肉鸡的肠胃,其能够在pH=2的环境下存活4 h以上,且能耐受0.15%~0.30%的胆盐,表现出强大的肠道定植能力[5]。非解乳糖链球菌在肉鸡中应用表现出保护肠道绒毛结构、调节免疫能力和抗氧化能力以及抑制病原菌的作用,进而起到保护肠道健康、提升机体抗病力和改善生长性能的效果[9-10]。然而,在一些饲粮中添加益生菌对肉鸡生长性能影响的研究中发现,低剂量益生菌并未对生长性能产生显著影响[11-12]。类似地,本试验结果发现,饲粮中添加非解乳糖链球菌对白羽肉鸡生长性能并无显著影响,尽管较高添加剂量的非解乳糖链球菌(如T4组)在各阶段的生长性能指标有所提高,但促生长效果未显著增强。分析原因主要有2点,一方面是非解乳糖链球菌的作用相对缓和,更偏向于维护长期的肠道稳定性和健康,对快速增重的短期作用较弱[13];另一方面可能是由于剂量设置与菌株的特性并未完全匹配,未来研究可考虑进一步优化剂量。

3.2 非解乳糖链球菌对白羽肉鸡屠宰性能和肉品质的影响

屠宰性能和肉品质是影响肉鸡生产经济效益和产品品质的关键指标。本试验发现,各组间白羽肉鸡屠宰率、半净膛率、全净膛率、胸肌率、腿肌率和腹脂率均无显著差异,这进一步表明在相对较短的养殖周期内,菌群的调节作用可能尚未充分累积到可以显著影响屠宰性能指标的程度。与CON组相比,T3组白羽肉鸡胸肌pH45 min显著降低,T3组和T4组胸肌pH24 h和失水率显著降低,各试验组胸肌剪切力均显著降低。这表明饲粮中添加非解乳糖链球菌可能在屠宰后影响了肌肉酸度,pH降低会导致肌肉在成熟过程中保持更好的水分,从而减少失水率,保持肉的多汁性和口感。这一效果在T3组和T4组更为显著,可能与益生菌添加剂量较高使得其代谢调节效应增强有关。研究发现,非解乳糖链球菌由于不分解乳糖,往往产生较少的乳酸产物,此外其在营养物质代谢上更多地依赖其他碳源,产生的代谢物较少呈酸性,进而能够改善肉鸡的代谢平衡,降低肌肉中酸性物质的堆积[14]。肉色能反映肉质的生理变化,a*值越高,肉的新鲜度越好;b*值越高,肉的抗氧化能力越弱[15]。本研究发现,T1组和T2组白羽肉鸡胸肌a*和b*值相较于CON组显著降低,这是由于非解乳糖链球菌通过增强抗氧化能力和调节脂质代谢,可能降低了肌肉氧化作用,进而降低了肌红蛋白的氧化水平,使得肉质颜色变浅[16]。

3.3 非解乳糖链球菌对白羽肉鸡抗氧化能力的影响

T-SOD和GSH-Px是反映机体抗氧化能力的重要指标,是机体中主要的抗氧化物酶,具有清除自由基的作用;T-AOC反映了机体的总体抗氧化水平;MDA则反映了畜禽机体内部氧化受损的程度[17]。姚先赐等[18]、李寒梅[19]和Xu等[20]研究发现,饲粮中添加益生菌有助于提高肉鸡肠道短链脂肪酸(SCFAs)含量,进而改善肠道健康,促进机体抗氧化水平的提升。本试验中,饲粮中添加非解乳糖链球菌对白羽肉鸡血清抗氧化指标具有积极作用,尤其是在T4组,表现出较高的T-AOC以及T-SOD和GSH-Px活性,同时MDA含量较低,表明抗氧化能力最强,且脂质过氧化程度最低。在肌肉抗氧化指标中,T4组同样表现较佳,表明该组在提升白羽肉鸡肌肉抗氧化能力、减少氧化应激和脂质过氧化损伤方面具有明显的效果。且随着试验鸡日龄的增加,这种正面效应更加明显。高剂量非解乳糖链球菌进入肠道后,有可能更快、更多地定植在肠道内,能够优化肠道微生物平衡,这种作用通过抑制有害菌生长、增加有益菌比例来增强肠道屏障功能,降低由病原菌引发的内毒素和代谢废物进入血液循环的几率,从而降低体内氧化应激,这可能也是提升肌肉品质的原因[16]。此外,非解乳糖链球菌在代谢过程中会产生SCFAs等有益代谢产物,这些代谢物在高剂量条件下更为丰富。SCFAs不仅有助于维持肠道的低pH环境,抑制有害菌生长,还在体内可能通过改善肠道吸收能力和整体营养代谢,为机体提供更加稳定和充足的营养供应,具有潜在的抗氧化作用[21]。研究还表明,SCFAs通过促进脂肪酸氧化、抑制脂肪合成、调控肌肉代谢基因表达以及提高糖原合成效率等方式减少肌肉中脂肪沉积并改善肉质的保水性,进而提升肉品质[22-23]。以上结果表明,非解乳糖链球菌通过“肠道菌群-SCFAs-宿主代谢”轴,在抗氧化能力和肉品质提升中发挥关键作用。

3.4 非解乳糖链球菌对白羽肉鸡肌肉炎性因子表达的影响

白细胞介素-1(IL-1)可分为白细胞介素-1α(IL-1α)和IL-1β 2种亚型,在炎症状态下,以IL-1β为主。炎症细胞分泌的IL-1β能够刺激滑膜细胞产生金属蛋白酶、酪蛋白酶和胶原酶,从而使软骨的胶原纤维变性、蛋白多糖分解;IL-1β亦能够刺激滑膜细胞产生前列腺素E2并诱导内皮细胞黏附分子的表达,使滑膜细胞与浸润性炎性细胞反应性增强,从而导致肌肉及骨骼产生炎症反应,影响肉质[24]。IL-6不仅可以加速蛋白质的降解,在全身炎症、肿瘤恶病质和肌肉炎症等病理过程中的肌肉损耗中也起到了推动性的作用[25]。TNF-α可以抑制脂肪酸酶活性、抑制脂肪酸合成并刺激脂肪分解等,且能显著提高肌肉总蛋白的降解率,其中肌原纤维蛋白的降解率提高了3倍[26]。IL-2是一种重要的细胞因子,在调节机体的免疫应答和免疫稳态中发挥关键作用,其编码的蛋白主要由活化的T细胞分泌[27]。IL-10是一种重要的抗炎性细胞因子,主要由调节性T细胞(Treg)、B细胞、巨噬细胞和树突状细胞等分泌,其在抑制炎症反应、维持免疫稳态方面发挥关键作用[28]。本试验发现,在21和42日龄时,饲粮中添加非解乳糖链球菌显著降低了白羽肉鸡胸肌IL-1β、IL-6和TNF-α的mRNA相对表达量,表明其对炎症反应有抑制作用。非解乳糖链球菌作为一种益生菌,在肠道内与其他微生物协同,形成微生态平衡,从而抑制有害菌的生长。在健康的肠道环境下,益生菌及其代谢产物可促进有益菌群的生长,同时抑制潜在致病菌的增殖,从而减少肠道内毒素的产生和吸收[29]。且由于肠道和全身免疫系统的联系,这一过程可能进一步降低外周组织(如胸肌)中的炎症水平,反映为IL-1β、IL-6、TNF-α的表达下调,进而改善肉品质[30]。肌肉组织不仅是运动系统的主要组成部分,同时也是免疫调节和炎症应答的重要靶器官。本试验中,与CON组相比,T1组和T3组在21日龄时以及T2组在42日龄时胸肌IL-2和IL-10 mRNA相对表达量显著降低,这表明益生菌在抑制炎症的同时,可能通过降低抗炎因子的过度表达来维持免疫系统的动态平衡,这种调节机制有助于机体避免过度的免疫抑制,从而维持健康的免疫功能。崔宇[31]研究表明,枯草芽孢杆菌可显著提高AA肉鸡脾脏IL-2的mRNA相对表达量,进而调控炎症反应,这与本试验研究结果存在差异,表明不同益生菌可能通过不同途径调控机体炎症反应。

4 结论

综上所述,在本试验条件下,白羽肉鸡饲粮中添加适宜剂量鸡源非解乳糖链球菌可通过增强抗氧化能力和调节炎症反应,进而提高肉品质和缓解氧化应激。

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