CN106070019A - 一种应用苹果酸在肉鸡初阶生产过程中弯曲菌的减除方法 - Google Patents
一种应用苹果酸在肉鸡初阶生产过程中弯曲菌的减除方法 Download PDFInfo
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Abstract
本发明提供了一种在肉鸡初级生产阶段弯曲菌的减除方法,能安全有效的降低鸡肉体内弯曲菌数量且不影响肉鸡生产。本发明利用苹果酸对肉鸡出栏前饮水进行处理。苹果酸添加入肉鸡日常饮水中。最佳抑菌条件为:所述苹果酸浓度为0.075%、饲喂方式为肉鸡出栏前饮水中持续添加1周。本发明的方法使饲喂丙酸后的肉鸡泄殖腔弯曲菌数量达到较低水平,同时不影响肉鸡的肠道健康、生产性能及肉品质。本发明较传统的饲喂抗生素防治方法更安全,更高效,且对肉鸡生产的影响较小。
Description
技术领域
本发明属于养殖场减除细菌的技术,特别是饲养过程中肉鸡弯曲菌携带量的减除方法。
背景技术
弯曲菌(Campylobacter),一种革兰阴性菌,是全世界细菌性腹泻病的主要诱因之一。特定血清型的弯曲菌还可作为格林-巴利综合症(Guillain-Barré syndrome,GBS)的前驱因子,对健康造成严重损害。欧盟2009年内报道了2017711例弯曲菌相关病例,而2013年增加到了214779例。与2006-2008年期间的数据相比,2014年美国该病原菌相关病例数量上升了13%。弯曲菌普遍分布于环境中,且定植的原始部位是盲肠和小肠。被污染的禽肉是该肠道病原菌的主要来源,肉鸡是该病原菌的天然宿主,被感染后其消化道内可携带大量弯曲菌,尤其在盲肠中的数量极为庞大,高达106-108CFU/g。养殖过程中肉鸡的弯曲菌肠道定植是后续胴体加工过程中弯曲菌交叉污染的主要原因,使弯曲菌通过禽肉生产链传播至零售环节成为可能。研究表明,通过在养殖环节减少弯曲菌在肉鸡体内的定植可以降低弯曲菌通过食物链传播至人类的风险,从而减少弯曲菌病例的发生,对社会公共卫生具有重要意义。
饲喂抗菌剂是降低肉鸡体内弯曲菌数量传统而有效的措施,主要抗菌剂有抗生素、微生物制剂、植物天然提取物及有机酸等。但抗生素的使用存在引发菌株耐药性和药物残留等安全隐患。据EFSA报道,从人类病例中获得的弯曲菌分离株对临床常见抗生素具有高耐药性。此外,来自食物及野鸡、鸡肉、猪和牛等动物源性的弯曲菌分离株对于喹诺酮类药物的抗性极高。为此欧盟和美国已禁止以非治疗性目的在动物饲料中添加抗生素。所以我们要寻找更安全有效 的减菌措施来替代目前并不完善的防控措施。在众多抗生素替代产品中,有机酸以其广谱性抑菌、天然无公害等优势吸引了研究者们的注意,已有文献证实有机酸作为饮水或饲料添加剂可以减少肉鸡弯曲菌及其他肠道病原菌的携带量,但也有研究表明,在日粮中添加有机酸后肉鸡的肠道生长、生产性能和肉品质会受到显著影响。消化器官生长发育的好坏决定着动物的生长快慢和健康,有机酸主要作用于家禽肠道,通常通过测定肠道的重量、长度及pH来评价其对消化器官的影响。评价肉鸡生产性能的指标主要有体重、平均日增重。GB102212-88规定了肉是由水分,蛋白质,脂肪,维生素及其他微量物质组成的复杂生物系统,肌肉中各种物质的种类、数量及其组成方式是肉品质的决定因素。因此测定肌肉常规成分可用于评价鸡肉口感与营养。
发明内容
本发明目的在于提供一种应用低浓度苹果酸在肉鸡初阶生产过程中弯曲菌的减除方法,能安全有效的降低肉鸡体内弯曲菌数量且影响肉鸡肠道、生产性能及肉品质。
为实现上述目的,本发明采用以下技术方案:
一种肉鸡生产初级阶段弯曲菌的减菌方法,利用苹果酸对肉鸡出栏前饮水进行处理,将饮水的pH调整至3.6~4.0。饮水pH为4.0(苹果酸终浓度为0.075%)。
上述方法具体步骤为:将苹果酸溶于无菌蒸馏水中配制成母液,添加于肉鸡饮水中。
优选地,饮水pH为4.0,所述苹果酸在饮水中的最终质量浓度为0.075%、酸化饮水饲喂时间为出栏前1~3周。
本发明还公开了苹果酸在制备肉鸡初阶生产过程中弯曲菌的减菌剂中的应用。所述应用具体是:在肉鸡出栏前饮水中添加苹果酸,将饮水的pH调整至3.6~4.0。 优选地,上述应用中所述苹果酸的质量浓度为0.075%,肉鸡初阶生产过程是指肉鸡出栏前1~3周。
本发明的有益效果是:本发明的方法使饮用苹果酸的肉鸡弯曲菌携带量达到较低水平,同时不影响鸡的肠道健康、生产性能及肉品质。本发明较传统的防控方法更安全,更高效,且对肉鸡生产的影响较小。由于有机酸具有一定腐蚀性,过量添加可能影响饲料及饮水的适口性,降低采食及饮水量,本发明所使用的苹果酸浓度较低,酸度范围合理,能够在有效降低肉鸡肠道弯曲菌载量的同时对肉鸡的生产性能不造成不利影响。另外,由于弯曲菌病是人兽共患病,处理或食用污染的鸡肉产品及其交叉污染物,是人感染弯曲菌的最重要途径,同时鸡携带弯曲菌的比例可达100%,而且鸡肠道内容物的含菌量可达109/克。本发明是专门针对肉鸡初级生产阶段,开展如何降低肠道弯曲菌携带量做的一系列研究,意在研发出针对于弯曲菌的最佳抑菌条件,旨在有效的降低肉鸡弯曲菌携带量,这不同于传统的抗菌剂治疗方法。
附图说明
图1是不同质量浓度苹果酸对弯曲菌抑制效率的比较;
图2是与苹果酸共培养20h后弯曲菌的扫描电镜图片;
图3是苹果酸对弯曲菌蛋白质外泄的影响;
图4是苹果酸对弯曲菌核酸渗漏的影响;
图5是苹果酸对肉鸡增重的影响。
具体实施方式
下面结合具体实施方式对本发明作进一步说明。
实施例1
体外致死临界浓度的筛选
首先对0.05%、0.10%、0.15%三个质量浓度的苹果酸对弯曲菌的抑制效率 进行比较。将培养状态相同的弯曲菌与不同质量浓度的苹果酸42℃共培养30h,弯曲菌起始浓度为5×107CFU/mL,每10h平板计数一次,绘制添加各浓度苹果酸后空肠弯曲菌11168菌株的生长曲线。结果表明,0.05%、0.1%、0.15%三个质量浓度的苹果酸中,在30h内可将对空肠弯曲菌11168菌株浓度降低至不可检测水平的只有0.15%质量浓度的苹果酸(见图1)。
实施例2
实际应用
为了判断方案的可行性,对养殖场肉鸡进行试验以验证苹果酸对肉鸡肠道中弯曲菌的作用效果。按商品肉鸡的饲养要求控制环境温度及湿度,入雏前1周,熏蒸消毒并将鸡笼及用具全部冲洗干净,全程定期消毒。采用笼养方式,通过塑料饮水器供水,自由釆食、饮水。饲养期间各组免疫程序相同。选用个体差异相对较小的5周龄肉鸡60只,随机分成三个组,苹果酸组、酒石酸组和对照组。两个处理组分别用苹果酸、酒石酸将肉鸡日常饮用水pH调为4.0(苹果酸质量浓度为0.075%,酒石酸的质量浓度为0.059%);对照组则供应养殖场日常饮水。有机酸饲喂3周,于有机酸饲喂当天及饲喂后的第1、2、3周采集各组肉鸡肛拭样品测定弯曲菌阳性率、携带量及肠道微生物菌落总数。
由表1和表2可知,苹果酸和酒石酸对肉鸡弯曲菌携带率无影响,但应用1周时苹果酸可使弯曲菌数量显著降低0.41个数量级(P=0.0139),抑菌率为61.10%;而酒石酸组与对照组无显著差异(P>0.05)。由表3可知,苹果酸和酒石酸应用各周对肉鸡肠道微生物区系无显著影响(P>0.05)。表明饲喂0.075%苹果酸不会影响肉鸡肠道微生态平衡,且可有效降低弯曲菌的携带量。
表1肉鸡泄殖腔弯曲菌阳性率计算结果(%)
表2肉鸡泄殖腔弯曲菌携带量测定结果(log10CFU/g)
表3肉鸡肠道菌落总数测定结果(log10CFU/g)
实施例3
对弯曲菌影响的测定
应用扫描电镜(SEM)对苹果酸组弯曲菌的细胞壁进行观察,具体操作方法如下:取800μL MH液体培养基,、加入100μL 1.5%(w/v)的苹果酸,以100μL的MH液体培养基代替作为对照。在CCDA固体培养基上复苏细菌,于42℃微需氧条件下培养24h后用MH液体培养基收获细菌,并用MH培养基调整弯曲菌的接种浓度为5×108CFU/mL。向配制好的有机酸溶液中加入稀释好的菌液100μL。42℃摇床,120rpm微需氧条件下培养。于20h取菌悬液做为待测样品,保证两实验组同时进行。4℃,5000rpm离心5min,弃上清。后续固定、脱水、样品干燥及扫描电镜(SEM)观察均在扬州大学测试中心完成。
同时通过紫外分光光度计对弯曲菌内容物外泄进行测定,前期苹果酸与弯曲菌的培养步骤同上,分别于10h、20h、30h,3500rpm离心10min,取上清液于紫外分光光度计260nm波长处测定核酸渗透;于280nm波长处测定菌体蛋白 质的泄漏。
结果显示,培养20h时苹果酸对弯曲菌细胞壁的形态产生了影响,并导致核酸与蛋白质的大量泄漏(见图2、3、4)。
实施例4
对肉鸡肠道及生产性能的影响
1肉鸡肠道总重、长度及pH值的测定
各组随机抽取体重接近均值的5只鸡进行解剖,剖腹取出肠道并将各肠段结合处结扎。将肠道进行称重,并测量十二指肠、空肠、回肠的长度。待肠道组织长度测定后,分别剪开空肠、盲肠的一端,挤出肠道内容物于灭菌的玻璃瓶中,按1∶8(w/v)加入去离子水稀释,玻璃棒搅拌均匀后测定其pH值。
2肉鸡生产性能的测定
体重和平均日增重的测定:
从各处理组饮用酸化饮水的当天起,按不同时间点测定每只鸡的体重,对试验鸡进行空腹称重,称重前晚22时断料、保持供水,次日早8时称重、记录,并计算平均日增重。计算方法如下:
平均日增重(ADG)=(试验末期平均体重-初期平均体重)/试验天数肌肉常规成分测定:
水分(冷冻干燥法):取出-20℃冰箱冷冻保存的肉样,在室温条件下解冻,置于玻璃平皿中用真空冷冻干燥机冷冻干燥,测定水分含量。冷冻干燥后的样品用研磨磨碎后,用于粗蛋白质(凯氏定氮法)、粗脂肪(索氏提取法)、粗灰分(高温灼烧法)测定,具体测定方法参考《饲料分析及饲料质量检测技术》,计算方法如下:
粗蛋白%=0.014×滴定中消耗盐酸的体积÷试样质量×盐酸标准溶液浓度 ×100×转换系数F;粗脂肪%=(接收瓶、沸石连同脂肪的质量-接收瓶和沸石的质量)×100/试样的质量;粗灰分%=(坩埚和灰分的质量-空坩埚的质量)×100/(坩埚和试样的质量-空坩埚的质量)。其中,粗蛋白%、粗灰分%、粗脂肪%的各测定指标的单位均为g。粗蛋白的测定中鸡肉的转换系数F=6.25。粗脂肪%的计算结果保留小数点后一位。
由表4可知,饲喂苹果酸的对肉鸡肠道总重、pH及长度均无显著影响(P>0.05)。由表5和图5可知,饲喂苹果酸对肉鸡体重及增重具有有利影响。由表6可知,饲喂苹果酸可以降低肉鸡死淘率,有利于肉鸡存活。由表7可知,饲喂苹果酸对肉鸡的肉品质无显著不利影响(P>0.05)。
表4肉鸡肠道总重、长度和pH的测定结果
表5肉鸡体重的测定结果(g)
表6肉鸡死淘率的统计结果(%)
表7肉鸡肌肉常规成分(%)
注:在表7中,水分指标是以鲜样为基础,粗灰分、粗蛋白和粗脂肪是以冻干样为基础。
结果表明,苹果酸可显著降低在肉鸡体内的弯曲菌数量,抑菌率达到61.10%,且对肉鸡的肠道微生态平衡和生长及生产性能均无显著不良影响,可以降低肉鸡死淘率,说明0.075%苹果酸应用于肉鸡养殖环节弯曲菌的防控是安全有效的。
Claims (7)
1.一种应用苹果酸在肉鸡初阶生产过程中弯曲菌的减除方法,其特征在于:用苹果酸对肉鸡出栏前饮水进行处理。
2.如权利要求1所述的方法,其特征在于:在肉鸡出栏前饮水中添加苹果酸,将其pH调整至3.6~4.0。
3.如权利要求2所述的方法,其特征在于:处理后饮水的pH值为4.0,苹果酸的终质量浓度为0.075%。
4.如权利要求2所述的方法,其特征在于:在肉鸡出栏前饮水中持续添加1~3周苹果酸。
5.苹果酸在制备肉鸡初阶生产过程中弯曲菌的减菌剂中的应用。
6.如权利要求5所述的应用,其特征在于:所述苹果酸的质量浓度为0.075%,肉鸡初阶生产过程是指肉鸡出栏前1~3周。
7.如权利要求5所述的应用,其特征在于:在肉鸡出栏前饮水中添加苹果酸,将饮水的pH调整至3.6~4.0。
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