Chinese Journal of Animal Nutrition >

2024 , Vol. 36 >Issue 12: 7687 - 7697

DOI: https://doi.org/10.12418/CJAN2024.656

RESEARCH PAPER

Effects of Compound Probiotic Preparation on Performance, Egg Quality, Serum Indexes, Nutrient Apparent Metabolic Rates and Fecal Pollutant Emissions of Laying Hens

  • YANG Bowen , 1 ,
  • DONG Tongchao 1 ,
  • WANG Jue 2 ,
  • WANG Juan 3 ,
  • CHEN Xiaolong 1 ,
  • LIU Huage 2 ,
  • WANG Xuejing , 2, * ,
  • XU Lijun , 4, * ,
  • CHEN Ligong 1 ,
  • WANG Xue 5 ,
  • WANG Chengmin 5 ,
  • XU Lili 6 ,
  • JIN Jingjing 7
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  • 1 Hebei Agricultural University, Baoding 071000, China
  • 2 Hebei Animal Husbandry and Veterinary Research Institute, Baoding 071030, China
  • 3 Shijiazhuang Engineering Vocational College, Shijiazhuang 050204, China
  • 4 Baoding Animal Husbandry Workstation, Baoding 071023, China
  • 5 Institute of Zoological Research, Guangdong Academy of Sciences, Guangzhou 510075, China
  • 6 Union Biotechnology (Shanghai) Co., Ltd., Shanghai 201400, China
  • 7 Hebei Huanshan Agricultural Development Co., Ltd., Baoding 074200, China
*WANG Xuejing, professor, E-mail: ;
XU Lijun, senior husbandry specialist, E-mail:

Received date: 2024-05-30

  Online published: 2024-12-12

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Abstract

The purpose of this experiment was to investigate the effects of compound probiotic preparation on the performance, egg quality, serum indexes, nutrient apparent metabolic rates and fecal pollutant emissions of laying hens, and to provide experimental basis for the application of compound probiotic preparation in layer production. A total of 288 fifty-weeek-old Jinghong laying hens with similar egg production rate and health were randomly selected and divided into four groups, with six replicates for each group and 12 hens for each replicate. Laying hens in control group were fed a corn-soybean meal type basal diet, and those in experimental groups were fed the control group’s diet supplemented with 100, 150 and 200 mg/kg of compound probiotic preparation (It consisted of Bacillus subtilis, Bacillus licheniformis, Clostridium butyricum and Enterococcus faecalis), respectively. The pre-feeding period was 7 d, and the formal test period was 84 d. The results showed as follows: 1) the laying rate of the 150 mg/kg group was significantly higher than that of the control group (P<0.05); the broken egg rate of the 200 mg/kg group was significantly lower than that of the control group (P<0.05), but the difference between the 150 and 200 mg/kg groups was insignificant (P>0.05). 2) The eggshell thickness of the 200 mg/kg group was significantly higher than that of the control group (P<0.05), and there was no significant difference between the 200 and 150 mg/kg groups (P>0.05). 3) Serum albumin content of the 150 mg/kg group was significantly higher than that of the other group (P<0.05); serum urea nitrogen content of the 200 mg/kg group was significantly lower than that of the control group (P<0.05), and there was no significant difference with that in the 150 mg/kg group (P>0.05). 4) Crude protein apparent metabolic rate of the 200 mg/kg group was significantly higher than that of the control group and 100 mg/kg group (P<0.05), but not significantly different from that of the 150 mg/kg group (P>0.05); calcium apparent metabolic rate of the 150 and 200 mg/kg groups was significantly higher than of the control group and 100 mg/kg group (P<0.05), but there was no significant difference between the 150 and 200 mg/kg groups (P>0.05). 5) The indole and 3-methylindole contents and ammonia emission in the feces of 150 and 200 mg/kg groups was significantly lower than those of the control group and 100 mg/kg group (P<0.05), and there was no significant difference between the 150 and 200 mg/kg groups (P>0.05). 6) The villus height of the duodenum of the 200 mg/kg group was significantly higher than that of the control group and 100 mg/kg group (P<0.05), and there was no significant difference between the 150 and 200 mg/kg groups (P>0.05). 7) The highest average profit per hen was recorded in the 150 mg/kg group, which was 7.40% higher than that of the control group. In conclusion, it is recommended that the additive amount of compound probiotic preparation in the diet of laying hens should be 150 mg/kg.

Key words： compound probiotic preparation; laying hens; performance; egg quality; fecal pollutant emissions

Cite this article

YANG Bowen , DONG Tongchao , WANG Jue , WANG Juan , CHEN Xiaolong , LIU Huage , WANG Xuejing , XU Lijun , CHEN Ligong , WANG Xue , WANG Chengmin , XU Lili , JIN Jingjing . Effects of Compound Probiotic Preparation on Performance, Egg Quality, Serum Indexes, Nutrient Apparent Metabolic Rates and Fecal Pollutant Emissions of Laying Hens[J]. Chinese Journal of Animal Nutrition, 2024 , 36(12) : 7687 -7697 . DOI: 10.12418/CJAN2024.656

益生菌能够保护动物胃肠道,促进其健康生长,并且通过其有益代谢产物影响动物的健康,减少环境污染,降低养殖成本,因此成为了近年来的研究热点。益生菌作为绿色、安全的饲料添加剂已被广泛应用于畜禽养殖中。研究显示,枯草芽孢杆菌具有提高蛋鸡生产性能、改善蛋品质、促进肠道健康发育、提高养分吸收的作用[1];地衣芽孢杆菌能够提高蛋鸡的生产性能、促进肠道健康,并能影响盲肠微生物组成,调节肠道微生态平衡[2];丁酸梭菌是一种主要代谢产物是丁酸的益生菌,其可以通过提高肠道屏障功能和免疫力改善蛋鸡的卵巢功能和生产性能[3];粪肠球菌可以改善蛋鸡盲肠微生物结构和血清生化指标[4]。复合益生菌制剂是由几种益生菌组合加工制成的制剂,能够发挥多种益生菌的正向作用,进而促进动物的生长和生产[5]。近年来有很多研究均表明复合益生菌制剂可以提高蛋鸡的生产性能、增强免疫力,但由于益生菌种类、组合方式及添加量等不同,复合益生菌制剂在动物体内产生的效果不同。本试验选择了一种主要成分为枯草芽孢杆菌、地衣芽孢杆菌、丁酸梭菌、粪肠球菌的复合益生菌制剂,探究其对蛋鸡生产性能、蛋品质、血清指标、养分表观代谢率、肠道形态和粪污排放的影响,以期为该复合益生菌制剂在蛋鸡生产中的应用提供试验支撑。

1 材料与方法

1.1 试验材料

复合益生菌制剂组成:枯草芽孢杆菌(6×109 CFU/g)、地衣芽孢杆菌(4×109 CFU/g)、丁酸梭菌(5×108 CFU/g)、粪肠球菌(1×109 CFU/g)。

1.2 试验设计与饲粮组成

试验方案经河北省畜牧兽医研究所实验动物伦理审查委员会批准(编号01)。饲养试验在河北省易县永合养鸡场进行。选取产蛋率相近、健康的50周龄京红蛋鸡288只,随机分为4组,每组6个重复,每个重复12只鸡。对照组采用玉米-豆粕型基础饲粮,该基础饲粮参照《鸡饲养标准》(NY/T 33—2004)配制(表1);试验组分别在对照组饲粮的基础上添加100、150和200 mg/kg的复合益生菌制剂。预试期7 d,正试期84 d。
表1 基础饲粮组成及营养水平(风干基础)

Table 1 Composition and nutrient levels of the basal diet (air-dry basis) %

原料Ingredients 含量Content 营养水平Nutrient levels2) 含量Content
玉米Corn 58.54 代谢能ME/(MJ/kg) 11.34
豆粕Soybean meal 24.86 粗蛋白质CP 17.22
麸皮Wheat bran 4.00 钙Ca 3.28
预混料Premix1) 3.00 总磷TP 0.65
豆油Soybean oil 1.00 有效磷AP 0.39
石粉Limestone 8.60 赖氨酸Lys 0.85
合计Total 100.00 蛋氨酸Mst 0.26

1)预混料为每千克饲粮提供 The premix provided the following per kg of the diet:VA 8 000 IU,VD3 3 600 IU,VE 21 IU,VK3 4.2 mg,VB1 3 mg,VB2 10.2 mg,叶酸 folic acid 0.9 mg,泛酸钙 calcium pantothenate 15 mg,烟酸 niacin 45 mg,VB6 5.4 mg,VB12 0.024 mg,生物素 biotin 0.15 mg,Cu 6.8 mg,Fe 66 mg,Zn 83 mg,Mn 80 mg,I 1 mg,Se 0.3 mg。

2)除代谢能和有效磷外,其他指标均为测定值。代谢能和有效磷根据中国饲料数据库(https://www.chinafeeddata.org.cn)计算。ME and AP were calculated values, while the others were measured values. ME and AP were calculated based on the Chinese Feed Database (https://www.chinafeeddata.org.cn).

1.3 饲养管理

蛋鸡采用立式笼养,试验期间通过湿帘、通风等手段控制鸡舍温度在26 ℃左右,相对湿度在55%;每日饲喂2次(8:00、15:00),自由采食和饮水;光照采用人工光照,每日16 h;按常规程序进行免疫。

1.4 检测指标及方法

1.4.1 生产性能

试验开始后,每天记录每个重复的产蛋数、蛋重、鸡死亡数、不合格蛋数,每周结料,统计耗料量。试验结束后计算每组的产蛋率、平均日采食量(ADFI)和料蛋比(F/E)。
产蛋率(%)=[产蛋总数/(鸡只数×统计天数)]×100;
平均日采食量(g)=总耗料重/(鸡只数×统计天数);
料蛋比=总耗料重/总蛋重;
不合格蛋率(%)=(不合格蛋数/产蛋总数)×100。

1.4.2 蛋品质

试验结束当天,每组随机选取30枚鸡蛋,标号,存放于4 ℃冰箱保存,并于收集当天进行蛋品质测定。使用游标卡尺测定鸡蛋长径和短径,计算蛋形指数;使用蛋壳强度测定仪(Orka Food Technology Co., Ltd.,以色列)测定蛋壳强度;使用蛋品质分析仪(Orka Food Technology Co., Ltd.,以色列)测定蛋白高度、蛋黄颜色、哈氏单位;使用电子天平称蛋重和蛋黄重,计算蛋黄比率;使用螺旋测微仪测定蛋壳厚度(每枚鸡蛋测量钝、中、锐3个部位,取平均值)。

1.4.3 养分表观代谢率

在饲养试验的最后3 d,以重复为单位每24 h收集1次排泄物(去除其中羽毛、饲粮等杂物),在-20 ℃冰箱中保存,以防止样品腐败变质。样品收集完毕后,将3 d的排泄物混合均匀,经过65 ℃烘干制成风干样品,粉碎过40目筛,制成待测样品封于密封袋中,在-20 ℃冰箱中保存待测。饲粮和排泄物中干物质含量参照GB/T 6435—2014测定,粗蛋白质含量参照GB/T 6432—2018(凯氏定氮法)测定,粗灰分含量参照GB/T 6438—2007测定,总磷含量参照GB/T 6437—2018(采用分光光度法)测定,钙含量参照GB/T 6436—2002(高锰酸钾滴定法)测定,酸不溶灰分含量参照GB/T 23742—2009测定。
养分表观代谢率计算公式如下:
某养分表观代谢率(%)=[1-(a/b)×(c/d)]×100。
式中:a为排泄物中该养分含量;b为饲粮中该养分含量;c为饲粮中酸不溶灰分含量;d为排泄物中酸不溶灰分含量。

1.4.4 血清指标

饲养试验最后1天禁食12 h,每重复随机选取1只鸡采集血液10 mL,常温条件下倾斜促凝管静置30 min,经1 500×g离心10 min,采集血清,分装至离心管中,将其置于-20 ℃保存待测。使用南京建成生物工程研究所生产的对应试剂盒测定血清中总蛋白(TP)、白蛋白(ALB)、球蛋白(GLB)、总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、尿素氮(UN)、葡萄糖(GLU)、免疫球蛋白A(IgA)、免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、白细胞介素-4(IL-4)、肿瘤坏死因子-α(TNF-α)、丙二醛(MDA)的含量和谷草转氨酶(AST)、谷丙转氨酶(ALT)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性以及总抗氧化能力(T-AOC)。

1.4.5 粪中污染物质排放

在饲养试验的最后3 d,以重复为单位每24 h收集1次排泄物(去除其中羽毛、饲粮等杂物),在-20 ℃冰箱中保存,以防止样品腐败变质。样品收集完毕后,将3 d的排泄物混合均匀后取样待测。
粪中吲哚和3-甲基吲哚(粪臭素)含量使用试剂盒测定,试剂盒均购自上海迈瑞尔生化科技有限公司。
粪中氨气排放量测定:每个重复取100 g新鲜鸡粪,置于500 mL磨口玻璃瓶中,于室温环境下发酵3 d。参照《环境空气和废气 氨的测定 纳氏试剂分光光度法》(HJ 533-2009)的方法使用纳氏试剂比色法测定。

1.4.6 小肠形态结构

饲养试验结束时,每个重复选取1只鸡空腹12 h后屠宰,迅速剖取小肠(十二指肠、空肠、回肠)样品,在4%的多聚甲醛溶液中固定。组织固定48 h后经脱水、透明后使用石蜡进行包埋,经苏木精-伊红染色制作常规石蜡切片,石蜡切片置于显微镜下观察。以10×10的倍数,在显微镜下测量十二指肠、空肠和回肠的绒毛高度和隐窝深度。从每个切片中选取3个视野,每个视野中选取5个绒毛或隐窝测量绒毛高度或隐窝深度,并计算绒隐比(绒毛高度/隐窝深度)。

1.4.7 经济效益

根据试验期间饲料的消耗量和饲料市场均价计算饲料成本,不计入死亡鸡耗料量。根据试验期间蛋鸡总产蛋量和鸡蛋市场均价计算收入,不计入不合格蛋。试验期间,基础饲粮单价为2.98元/kg,所用复合益生菌制剂单价为135元/kg,鸡蛋单价为11.58元/kg,未考虑人工、水电等其他成本。平均每只鸡利润计算公式为:
平均只利润=只总产蛋重×鸡蛋单价-只总耗料量×饲料单价。

1.5 数据统计分析

试验数据用SPSS 22.0统计软件进行单因素方差分析,并采用Duncan氏法进行多重比较,数据用“平均值±标准差”表示,P<0.05为差异显著。

2 结果与分析

2.1 复合益生菌制剂对蛋鸡生产性能的影响

由表2可知,150 mg/kg组的产蛋率显著高于对照组(P<0.05),其他组之间差异不显著(P>0.05);200 mg/kg组的不合格蛋率显著低于对照组(P<0.05),其他组之间差异不显著(P>0.05);平均蛋重、平均日采食量和料蛋比各组之间均无显著差异(P>0.05)。图1为产蛋率随时间变化的折线图,从图中可以看出,从第4周开始,添加复合益生菌制剂的试验组的产蛋率出现高于对照组的趋势,其中150 mg/kg组变化最明显。
表2 复合益生菌制剂对蛋鸡生产性能的影响

Table 2 Effects of compound probiotic preparation on performance of laying hens

项目
Items		 复合益生菌制剂添加量
Compound probiotic preparation additive amount/(mg/kg)		 P值
P-value
0 100 150 200
产蛋率Laying rate/% 89.94±2.52b 91.50±0.54ab 92.61±1.58a 91.64±1.19ab 0.019
平均蛋重Average egg weight/g 58.57±0.90 57.82±1.17 59.16±0.79 58.21±0.52 0.112
平均日采食量ADFI/g 113.44±2.65 111.32±2.94 114.14±4.28 113.41±3.43 0.519
料蛋比F/E 2.16±0.06 2.11±0.11 2.13±0.07 2.13±0.07 0.701
不合格蛋率Broken egg rate/% 6.47±1.82a 4.88±2.15ab 4.85±1.45ab 3.06±1.44b 0.041

同行数据肩标不同小写字母表示差异显著(P<0.05),无字母或相同字母表示差异不显著(P>0.05)。表3至表7同。

Data in the same row labeled with different lowercase letters on the shoulder indicate significant difference (P<0.05), and labeled with no letter or the same letter on the shoulder indicate non-significant difference (P>0.05). The following Tables 3 to 7 are the same.

图1 产蛋率随时间变化的折线图

Fig.1 Broken line graph of laying rate with time

2.2 复合益生菌制剂对蛋鸡蛋品质的影响

由表3可知,各组鸡蛋的蛋形指数、蛋壳强度、蛋白高度、哈氏单位、蛋黄比率均无显著差异(P>0.05);100 mg/kg组的蛋黄颜色较对照组以及150和200 mg/kg组显著下降(P<0.05),其他组之间差异不显著(P>0.05);200 mg/kg组的蛋壳厚度较其他组显著提高(P<0.05),其他组之间差异不显著(P>0.05)。
表3 复合益生菌制剂对蛋鸡蛋品质的影响

Table 3 Effects of compound probiotic preparation on egg quality of laying hens

项目
Items		 复合益生菌制剂添加量
Compound probiotic preparation additive amount/(mg/kg)		 P值
P-value
0 100 150 200
蛋形指数Shape index 1.32±0.04 1.34±0.04 1.33±0.04 1.33±0.04 0.612
蛋壳强度Eggshell strength/N 31.59±5.12 30.00±6.07 31.24±6.05 31.20±7.14 0.767
蛋白高度Albumen height/mm 5.10±1.17 4.94±1.16 4.67±1.06 4.83±0.70 0.444
蛋黄颜色Yolk color 7.31±0.80a 6.28±1.71b 6.97±0.84a 7.30±0.71a 0.002
哈氏单位Haugh unit 70.03±10.45 68.23±10.11 67.26±9.60 68.68±7.66 0.736
蛋黄比率Yolk percentage/% 28.02±2.20 28.43±2.07 27.64±2.05 28.49±1.87 0.354
蛋壳厚度Eggshell thickness/mm 0.33±0.02b 0.33±0.22b 0.33±0.02b 0.34±0.03a 0.031

2.3 复合益生菌制剂对蛋鸡血清指标的影响

由表4可知,150 mg/kg组的血清ALB含量较对照组和100 mg/kg组显著提高(P<0.05),其他试验组与对照组差异不显著(P>0.05);200 mg/kg组的血清UN含量显著低于对照组和100 mg/kg(P<0.05),与150 mg/kg组无显著差异(P>0.05);其他血清指标各组之间无显著差异(P>0.05)。
表4 复合益生菌制剂对蛋鸡血清指标的影响

Table 4 Effects of compound probiotic preparation on serum indexes of laying hens

项目
Items		 复合益生菌制剂添加量
Compound probiotic preparation additive amount/(mg/kg)		 P值
P-value
0 100 150 200
总蛋白TP/(g/L) 29.24±3.46 29.36±3.54 32.79±4.76 30.63±2.96 0.473
白蛋白ALB/(g/L) 17.01±1.21b 17.68±2.02b 20.77±2.46a 19.09±3.08ab 0.048
球蛋白GLB/(g/L) 12.23±2.60 11.67±2.30 11.07±1.27 11.55±1.54 0.082
谷丙转氨酶ALT/(U/L) 37.09±7.09 36.11±6.00 33.06±7.79 35.03±7.72 0.079
谷草转氨酶AST/(U/L) 143.62±2.62 145.99±20.82 157.65±18.88 155.59±7.22 0.283
总胆固醇TC/(mmol/L) 5.55±0.51 4.24±1.06 5.26±0.89 4.88±0.99 0.095
甘油三酯TG/(mmol/L) 10.95±0.40 10.57±0.77 11.03±0.40 11.03±0.23 0.340
高密度脂蛋白胆固醇HDL-C/(mmol/L) 1.79±0.17 1.32±0.39 1.71±0.30 1.54±0.35 0.076
低密度脂蛋白胆固醇LDL-C/(mmol/L) 2.54±0.33 1.75±0.61 2.33±0.56 2.12±0.64 0.109
尿素氮UN/(mmol/L) 6.94±1.86a 6.77±1.46a 4.78±2.38ab 4.14±1.44b 0.032
葡萄糖GLU/(mmol/L) 11.57±0.52 12.02±0.26 11.96±0.77 11.81±0.31 0.482
免疫球蛋白G IgG/(g/L) 4.14±0.07 4.18±0.17 4.21±0.09 4.22±0.07 0.566
免疫球蛋白A IgA/(g/L) 2.20±0.03 2.22±0.09 2.26±0.06 2.25±0.04 0.335
免疫球蛋白M IgM/(g/L) 1.61±0.04 1.62±0.07 1.64±0.07 1.64±0.04 0.779
超氧化物歧化酶SOD/(U/L) 122.10±11.13 120.52±4.38 113.16±0.84 115.89±13.19 0.364
丙二醛MDA/(nmol/L) 4.38±0.77 4.48±0.31 4.89±0.19 4.85±0.93 0.399
总抗氧化能力T-AOC/(U/L) 9.22±0.96 9.07±0.36 8.49±0.06 8.62±1.21 0.395
谷胱甘肽过氧化物酶GSH-Px/(U/L) 1 067.78±91.30 1 045.69±9.94 1 018.61±20.66 1 019.55±86.01 0.548
肿瘤坏死因子-α TNF-α/(pg/mL) 14.67±2.76 14.81±0.86 16.85±0.79 15.98±2.93 0.259
白细胞介素-4 IL-4/(pg/mL) 34.10±5.52 32.95±2.05 31.23±1.55 31.08±5.28 0.521

2.4 复合益生菌制剂对蛋鸡养分表观代谢率的影响

由表5可知,200 mg/kg组的粗蛋白质表观代谢率显著高于对照组和100 mg/kg组(P<0.05),与150 mg/kg组无显著差异(P>0.05);150和200 mg/kg组的钙表观代谢率均显著高于对照组和100 mg/kg组(P<0.05),这2组之间无显著差异(P>0.05);其他养分的表观代谢率各组之间无显著差异(P>0.05)。
表5 复合益生菌制剂对蛋鸡养分表观代谢率的影响

Table 5 Effects of compound probiotic preparation on apparent metabolic rates of nutrients of laying hens %

项目
Items		 复合益生菌制剂添加量
Compound probiotic preparation additive amount/(mg/kg)		 P值
P-value
0 100 150 200
干物质DM 91.35±1.03 90.95±0.85 90.86±0.55 91.20±0.85 0.095
粗灰分Ash 41.98±6.89 39.25±5.71 38.65±3.72 43.77±4.85 0.095
粗蛋白质CP 60.39±2.04b 59.81±8.48b 67.54±7.10ab 68.30±13.06a 0.038
钙Ca 40.38±9.86b 37.97±7.65b 48.25±2.17a 51.33±1.31a 0.001
总磷TP 44.20±8.07 42.32±2.92 43.33±14.10 45.01±4.94 0.938

2.5 复合益生菌制剂对蛋鸡粪中污染物质排放的影响

由表6可知,蛋鸡饲粮中添加复合益生菌制剂显著影响了蛋鸡粪中污染物质的排放量(P<0.05);150和200 mg/kg组粪中氨气排放量显著低于对照组(P<0.05),3个试验组之间无显著差异(P>0.05);150和200 mg/kg组粪中3-甲基吲哚含量显著低于对照组和100 mg/kg组(P<0.05),这2组之间差异不显著(P>0.05);粪中吲哚含量也有相似的趋势,150和200 mg/kg组显著低于对照组和100 mg/kg组(P<0.05),这2组之间差异不显著(P>0.05)。
表6 复合益生菌制剂对蛋鸡粪中污染物质排放的影响

Table 6 Effects of compound probiotic preparation on pollutant emissions from laying hen feces

项目
Items		 复合益生菌制剂添加量
Compound probiotic preparation additive amount/(mg/kg)		 P值
P-value
0 100 150 200
氨气Ammonia/(mg/m3) 2 334.46±1 131.14a 1 701.69±378.99ab 1 308.47±207.03b 1 198.87±142.88b 0.028
3-甲基吲哚3-methylindole/(pg/mL) 570.56±9.93a 537.91±6.06a 505.94±6.49b 504.26±18.54b 0.001
吲哚Indole/(pg/mL) 3 635.56±155.30a 3 047.73±69.00a 2 210.50±15.73b 2 320.90±90.30b 0.001

2.6 复合益生菌制剂对蛋鸡小肠形态结构的影响

各组蛋鸡具代表性小肠组织切片见图2。由表7可知,200 mg/kg组蛋鸡十二指肠的绒毛高度显著高于对照组和100 mg/kg组(P<0.05),与150 mg/kg组差异不显著(P>0.05);其他小肠形态结构指标各组之间无显著差异(P>0.05)。
图2 各组蛋鸡小肠具代表性组织学切片

蛋鸡十二指肠组织切片的标尺为500 μm,空肠组织切片的标尺为100 μm,回肠组织切片的标尺为100 μm。

Fig.2 Representative histological section of small intestine of laying hens in each group

Scale 500 μm for histological sections of the duodenum, scale 100 μm for histological sections of the jejunum, and scale 100 μm for histological sections of the ileum.

表7 复合益生菌制剂对蛋鸡小肠形态结构的影响

Table 7 Effects of compound probiotic preparation on small intestinal morphologic structure of laying hens

项目
Items		 复合益生菌制剂添加量
Compound probiotic preparation additive amount/(mg/kg)		 P值
P-value
0 100 150 200
十二指肠Duodenum
绒毛高度Villus height/μm 1 210.74±214.50b 1 186.41±267.39b 1 298.43±145.78ab 1 456.50±59.93a 0.012
隐窝深度Crypt depth/μm 223.47±57.03 256.48±69.95 252.30±39.61 258.38±81.26 0.586
绒隐比V/C 5.71±1.54 4.85±1.53 5.30±1.18 6.22±2.14 0.299
空肠Jejunum
绒毛高度Villus height/μm 955.61±238.51 904.26±164.26 1 034.00±204.91 1 017.26±92.31 0.381
隐窝深度Crypt depth/μm 172.04±31.34 178.92±21.50 184.32±29.47 169.98±35.64 0.698
绒隐比V/C 5.70±1.69 5.05±0.74 5.69±1.23 6.26±1.64 0.296
回肠Ileum
绒毛高度Villus height/μm 905.60±153.92 864.39±47.52 871.09±130.87 941.02±86.24 0.410
隐窝深度Crypt depth/μm 178.06±33.04 155.62±47.61 143.56±26.19 156.89±33.34 0.208
绒隐比V/C 5.29±1.58 5.96±1.58 6.19±1.11 6.13±0.76 0.401

2.7 复合益生菌制剂对蛋鸡养殖经济效益的影响

由表8可知,只考虑饲料成本的情况下,各试验组的平均只利润均高于对照组,其中以150 mg/kg组最高,比对照组提高了7.40%。
表8 试验期间各组蛋鸡的经济效益

Table 8 Economic efficiency of laying hens in each group during experimental period

项目
Items		 复合益生菌制剂添加量
Compound probiotic preparation additive amount/(mg/kg)
0 100 150 200
饲料单价Feed unit-price/(元/kg) 2.98 2.99 3.00 3.01
饲料消耗总量Total feed consumption/(kg/只) 9.53 9.35 9.59 9.53
鸡蛋单价Egg unit-price/(元/kg) 11.58 11.58 11.58 11.58
鸡蛋总产量Total egg production/(kg/只) 4.42 4.44 4.60 4.48
平均只利润Average profit per hen/(元/只) 22.84 23.47 24.53 23.24

3 讨论

3.1 复合益生菌制剂对蛋鸡生产性能的影响

本研究使用的复合益生菌制剂主要由枯草芽孢杆菌、地衣芽孢杆菌、丁酸梭菌、粪肠球菌4种益生菌复合而成。目前,益生菌对动物生产性能影响的研究较多。Prazdnova等[6]将枯草芽孢杆菌KATMIRA1933复合制剂添加到蛋鸡饲粮中,发现其可以显著提高蛋鸡的产蛋率和蛋品质。Guo等[7]研究发现,饲粮中长期添加枯草芽孢杆菌CGMCC 1.921可以显著提高蛋鸡的饲料转化率。Chen等[8]研究发现,饲粮中添加5×108 CFU/kg的枯草芽孢杆菌可以显著提高蛋鸡的产蛋量。Wang等[9]研究发现,饲粮中添加地衣芽孢杆菌可以改善蛋鸡肠道屏障功能,并调节生殖激素分泌,从而提高了产蛋性能。Huang等[10]研究发现,饲粮中添加107 CFU/kg的丁酸梭菌会降低产蛋率,但添加107和108 CFU/kg的丁酸梭菌可以提高平均蛋重。Zhang等[11]在蛋鸡饲粮中添加3.75×108和7.5×108 CFU/kg的粪肠球菌,发现其可以提高蛋鸡的产蛋率,降低料蛋比。综合以上报道,本研究所使用的复合益生菌制剂由多种益生菌共同发挥作用,改善肠道发育,提高养分吸收,从而提高了蛋鸡的生产性能。150 mg/kg组的生产性能和200 mg/kg组差异不显著而优于100 mg/kg组,可能是由于较高的添加量使益生菌更易定植和发挥作用。

3.2 复合益生菌制剂对蛋鸡蛋品质的影响

蛋品质受肠道吸收养分能力的影响。本研究发现,蛋鸡饲粮中添加200 mg/kg的复合益生菌制剂显著提高了蛋壳厚度,降低了不合格蛋率。Wang等[1]研究发现,生产后期蛋鸡饲粮中添加枯草芽孢杆菌可以显著提高蛋鸡肠道对钙的吸收能力,从而提高蛋壳强度、蛋壳厚度、蛋壳重量、蛋壳比例和蛋壳钙含量。Lei等[12]报道,蛋鸡饲粮中添加不同水平的地衣芽孢杆菌可以显著提高鸡蛋的蛋壳厚度。Yang等[13]使用枯草芽孢杆菌和地衣芽孢杆菌复合剂添加到产蛋后期蛋鸡饲粮中,发现其可以显著提高鸡蛋的蛋壳强度。Wang等[3]添加1×109 CFU/kg的丁酸梭菌到蛋鸡饲粮中,发现其显著提高了蛋黄特性、蛋白高度和哈氏单位。Zhan等[14]的研究也发现,随着蛋鸡饲粮中丁酸梭菌添加量的增加,鸡蛋产量、鸡蛋质量和蛋壳强度呈二次曲线增加,以5×104 CFU/kg组的增加量最大。Song等[4]报道,蛋鸡饲粮中添加微囊化的粪肠球菌可以显著提高蛋壳强度,对蛋壳厚度的影响不显著。以上研究结果均与本试验结果相似。益生菌对蛋鸡蛋壳品质的改善可能与其提高肠道健康和促进肠道中钙的吸收有关。此外,益生菌的添加还可能会引起肠道微生物群的变化[15]。肠道微生物群的变化可能通过泄殖腔影响输卵管微生物群的平衡[16],这可能会改变有机基质降解微生物组分的组成,进而影响蛋壳强度和蛋壳厚度[17]。

3.3 复合益生菌制剂对蛋鸡血清指标的影响

益生菌可以通过调节肠道健康影响机体的代谢,改善血清指标。本研究发现,蛋鸡饲粮中添加150和200 mg/kg的复合益生菌制剂均可以显著提高蛋鸡血清中ALB含量,显著降低UN含量。ALB主要作用是维持机体营养与渗透压,其含量高低反映机体的营养状态。UN是机体蛋白质代谢的主要终末产物,其含量高低反映机体对蛋白质的利用程度。Abd El-Hack等[18]研究发现,在玉米干酒糟及其可溶物(DDGS)型饲粮中添加枯草芽孢杆菌可以显著提高蛋鸡血清中ALB含量,降低氨的含量。Salem等[19]报道,蛋鸡饲粮中添加地衣芽孢杆菌和枯草芽孢杆菌复合制剂显著提高了蛋鸡血清中TP、ALB和GLB含量。Yang等[20]在肉鸡中的试验表明,饲粮中添加丁酸梭菌可以在第39天显著提高其血清中ALB含量,降低第21天的血清中UN含量。Zou等[21]使用由枯草芽孢杆菌、丁酸梭菌和粪肠球菌制成的益生菌复合制剂饲喂肉鸡,发现其可以显著提高肉鸡血清中ALB和TP的含量。以上研究结果均与本试验结果相似,复合益生菌制剂提高蛋鸡血清中ALB含量、降低UN含量可能是因为其通过提高肠道对蛋白质的吸收实现的。此外,已有研究显示复合益生菌制剂还有提高蛋鸡血清抗氧化能力和免疫力的作用[22-23],但本研究中未发现这些作用,可能与添加量和动物的生理状态有关。

3.4 复合益生菌制剂对蛋鸡养分表观代谢率和粪污排放的影响

鉴于上述生产性能、蛋品质和血清指标的差异,本研究进一步对复合益生菌制剂对蛋鸡养分表观代谢率和粪污排放进行了研究。本试验发现,复合益生菌制剂促进了蛋鸡肠道对蛋白质和钙的吸收,并减少了粪中粪臭素和吲哚含量和氨气排放量。Souza等[24]研究发现,饲粮中添加枯草芽孢杆菌PB6可以显著提高蛋鸡干物质、粗蛋白质和能量的利用率。Salem等[19]报道,饲粮中添加地衣芽孢杆菌和枯草芽孢杆菌复合制剂显著提高了蛋鸡的粗蛋白质和粗脂肪利用率。同样的,Abd El-Hack等[18]的研究发现,在玉米DDGS型饲粮中添加枯草芽孢杆菌可以显著降低蛋鸡粪便中的氮含量,并且随着玉米DDGS使用量的增加,这种降低氮排放的效果更加明显。Zou等[21]使用由枯草芽孢杆菌、丁酸梭菌和粪肠球菌制成的益生菌复合制剂饲喂肉鸡,发现其显著降低了粪便中硫化氢和氨气的排放。综合上述文献和本试验结果,150和200 mg/kg复合益生菌组粪中粪臭素和吲哚含量和氨气排放量降低可能是因为其促进了蛋白质的吸收,也可能与肠道内环境改善有关。
在钙的利用率方面,Wang等[1]研究发现,蛋鸡饲粮中添加2×109 CFU/kg枯草芽孢杆菌显著提高了钙和磷的利用率。据报道,枯草芽孢杆菌可降低老化蛋鸡肠道消化液的pH,增加其肠道吸收表面,从而提高钙的吸收率[25]。肠道低pH有利于钙的溶解,从而提高钙的消化和吸收[26]。钙结合蛋白1(CALB1)是十二指肠和上部空肠吸收钙的跨细胞主动转运过程的主要组成部分[27]。枯草芽孢杆菌有助于提高十二指肠中CALB1的相对表达水平,增加细胞内扩散速率和总胞浆钙浓度,从而实现提高钙利用率,增强蛋壳品质[1]。这与本研究中蛋品质部分结果相对应,可能是本研究所使用的复合益生菌制剂提高蛋壳厚度的原因之一。

3.5 复合益生菌制剂对蛋鸡肠道发育的影响

益生菌通过其竞争作用和分泌有益代谢产物改善肠道内环境。本研究中,饲粮中添加200 mg/kg复合益生菌制剂显著提高了蛋鸡十二指肠的绒毛高度,这可能是粗蛋白质和钙表观代谢率提高的原因。据报道,在饲粮中添加枯草芽孢杆菌可以增加蛋鸡小肠的绒毛高度和绒隐比[1]。吸收面积的增加和肠道形态的改善可能是复合益生菌制剂提高钙利用率,并提高蛋壳厚度的另一个原因。Yang等[20]研究发现,饲粮中补充丁酸梭菌降低了肉鸡空肠的隐窝深度,增加了绒毛高度与绒隐比。丁酸梭菌带来的有益效果可能部分归因于丁酸梭菌产生的丁酸盐促进了肠上皮的生长[28]。Pan等[2]报道,饲粮中添加地衣芽孢杆菌DSM5749改善了蛋鸡肠道形态(绒毛高度、隐窝深度和绒隐比)。Wang等[29]将丁酸梭菌和粪肠球菌复合益生菌制剂补充到仔猪饲粮中,发现该复合益生菌制剂增加了空肠绒毛高度和绒隐比,同时降低了空肠隐窝深度。上述报道均与本试验结果相似。综合本试验其他部分的结果,说明复合益生菌制剂可以通过提高蛋鸡肠道表面积来提高粗蛋白质和钙的表观代谢率,从而提高生产性能和蛋壳厚度,减少粪污排放。

4 结论

饲粮中添加复合益生菌制剂可提高血清中ALB含量,降低UN含量,改善小肠组织形态,提高粗蛋白质和钙的表观代谢率,从而提高生产性能和蛋壳厚度,降低不合格蛋率,并且可以减少粪中污染物质的排放。综合各项指标和经济效益,蛋鸡饲粮中复合益生菌制剂的推荐添加量为150 mg/kg。

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