天然沸石对肉鸡的营养效应与有害物减控机制研究
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摘要
天然沸石是一种性质独特、用途广泛的功能性材料,在生命科学领域中极具应用潜力。本研究以天然斜发沸石为试验材料,采用化学和X射线衍射法对其原料矿粉和改性产品品位及在体外模拟消化道pH环境中的稳定性进行分析评价；比较不同品位与粒度以及改性天然斜发沸石对肉鸡的营养和其它生物学效应；从肉鸡生产性能、血清二胺氧化酶与一氧化氮、胸腺细胞凋亡等方面研究沸石对产气荚膜梭菌感染所致负面影响的缓解作用,并探索相关机制；从肉鸡生产性能、铅在肌肉中的残留、血清HSP70的0D值、细胞凋亡相关基因的表达等指标着手,探讨天然斜发沸石对铅毒性的减控作用及相关机制。
1、天然沸石的品位及稳定性评价
采集两种天然沸石样品,进行改性处理和模拟消化道pH环境以及加NH4+处理,采用X射线衍射、阳离子交换容量(吸铵值)和电镜扫描进行测定分析。结果表明：样品I的主成分为斜发沸石,含量达50%以上,其杂质主要是长石和水云母,并含有部分闪石和二氧化硅,化学成分主要是Si和Al,并含有一定量的Ca、Fe、K、Mg、Na;样品Ⅱ的斜发沸石含量较低,杂质主要是长石和蒙脱石。经模拟消化道环境处理后,样品I斜发沸石的含量稍有下降,Pb含量分别下降了39.2%和36.8%；样品II斜发沸石含量未改变,但出现了海泡石。改性提高了沸石的阳离子交换容量；结果提示试验所用沸石具有良好的稳定性。
2、不同品位及改性沸石对肉鸡生产性能、养分表观代谢率、血液生化指标的影响
将504只1日龄AA肉鸡随机分为四个处理,每个处理6个重复,分别饲喂对照日粮,对照日粮添加5%普通沸石、高品位沸石和高品位改性沸石的试验日粮,研究不同品位沸石及改性沸石对肉鸡生产性能、养分表观消化率、血液生化指标的影响。结果表明：添加沸石后肉鸡后期的体增重分别比对照组提高了2.05%、7.62%(P<0.05)和4.73%(P<0.05),饲料转化率改善了0.98%、5.39%(P<0.05)和3.43%(P<0.05)；高品位和改性高品位沸石使肉鸡全期的体重提高了4.10%和2.41%,饲料转化率均改善1.56%；但添加沸石降低了肉鸡前期体重,而提高了饲料转化率(P<0.05)。与对照组相比,三种沸石使饲料干物质的表观消化率分别提高了4.05%、5.86%及3.03%,粗蛋白消化率分别提高了2.25%、6.16%和11.410%,能量消化率分别提高了3.59%、7.60%和7.16%；但未能提高钙的消化率,高品位和改性高品位沸石提高了磷的消化率；除蛋氨酸外,添加沸石提高了其他氨基酸的消化率,分别使总必需氨基酸表观消化率提高了2.69%、4.79%和4.53%(P>0.05),总非必需氨基酸表观消化率提高了2.77%、7.56%和6.080%(P>0.05),总氨基酸表观消化率提高了2.74%、6.1l%和5.26%(P>0.05)；改性高品位沸石降低了肉鸡粪便中的水分含量；沸石对肉鸡血液生化指标无明显影响。
3、不同品位及改性沸石对肉鸡肠道消化酶活性、DNA与RNA含量及黏膜形态的影响
就普通沸石、高品位沸石和高品位改性沸石对肉鸡肠道粘膜中的RNA及DNA含量、消化酶活性和形态结构的影响进行了测定分析。结果表明,日粮中分别添加5%普通沸石、高品位沸石和高品位改性沸石,可提高肉鸡前期和后期肠道粘膜中RNA含量及后期空肠粘膜中的DNA含量；普通和高品位沸石使肉鸡前期空肠粘膜中的DNA含量有所提高(P>0.05),但均降低了前期回肠粘膜中的DNA含量(P>0.05);对肉鸡后期肠道粘膜中的DNA含量则影响不一。饲料中添加三种沸石分别使肉鸡前期肠道淀粉酶和蛋白酶活性提高了23.4%、8.3%、15.9%和17.4%、17.2%、2.90%,后期蛋白酶活性提高了4.0%、25.2%、22.8%(P>0.05),普通沸石使肉鸡后期内源淀粉酶活性提高了12.8%,但高品位沸石和改性沸石却使其分别下降了25.1%和14.1%(P>0.05)。结果提示,沸石具有改善肉鸡肠道粘膜形态的功能。
4、不同粒度沸石对肉鸡生产性能、肠道菌群及组织重金属残留的影响以及沸石在不同料型饲料中的应用效果
在饲料中分别添加三种不同粒度的沸石粉,研究其对肉鸡生产性能、肠道菌群及肝脏与肌肉中重金属残留量的影响及沸石在不同料型饲料中的应用效果。结果表明,与对照组相比,在饲料中添加5%三种不同粒度的沸石均未影响肉鸡增重,但改善了饲料转化率(P<0.05),降低了死淘率；粗、中、细三种粒度沸石使肉鸡肌肉中的铅含量分别降低了10.06%、43.02%、39.11%,肝脏中砷含量分别下降了34.00%、29.85%、16.12%,粗、中粒度沸石使肝脏中铅含量分别降低了32.61%、24.78%,沸石对肌肉中的砷含量影响不大；沸石降低了血氨浓度及空肠中的大肠杆菌数,使空肠中的乳酸菌数有所提高。
饲喂颗粒饲料的肉鸡生产性能明显优于粉料(P<0.05)。颗粒饲料中添加沸石对肉鸡前期生产性能无影响,但在粉料中添加则显著降低了肉鸡生长速度、采食量和饲料转化率(P<0.05)。后期在颗粒饲料中添加沸石则降低了肉鸡生长速度(P<0.05)和饲料转化率(P>0.05),但不影响采食量；粉料后期对增重无影响,但明显降低了采食量和料重比(P<0.05)。颗粒料和粉料中添加沸石均不影响肉鸡全期增重,但颗粒料的饲料转化率有所上升(P>0.05),而粉料则明显降低(P<0.05)。
5、天然沸石缓解产气荚膜梭茵对肉鸡的有害影响及相关机制研究
384只1日龄的AA肉鸡随机分为4个处理,每个处理6个重复,分别为对照组(饲喂基础日粮)、肉鸡感染产气荚膜梭菌组(饲喂基础日粮)、感染产气荚膜梭菌肉鸡饲喂添加杆菌肽锌(40mg.kg-1)日粮和感染产气荚膜梭菌肉鸡饲喂添加5%沸石日粮,研究天然沸石缓解产气荚膜梭菌对肉鸡的有害影响及相关机制。结果表明：沸石明显提高了感染产气荚膜组肉鸡前期的饲料转化率(P<0.05),但对免疫器官指数和血清各生化指标均影响不明显(P>0.05)。与感染组相比,饲料中添加沸石使肉鸡前期血清SOD极显著提高(P<0.01),沸石缓解了产气荚膜梭菌对肉鸡造成的氧化损伤；抗生素也有相同效果。产气荚膜梭菌感染明显提高了肉鸡前期NO和后期DAO;沸石显著降低了肉鸡前期血清NO(P<0.01)和后期DAO(P<0.05),沸石可缓解产气荚膜梭菌对肠道的损伤；抗生素亦具同样作用。沸石和抗生素对肉鸡血清T3、T4、IgA和IgG均无影响(P>0.05)；但添加沸石可减少产气荚膜梭菌导致的胸腺细胞死亡数,提高活细胞数量(P>0.05)。
6、沸石对感染产气荚膜梭菌肉鸡生产性能、免疫器官及屠宰性能的影响
本试验设5个处理,分别为对照组(饲喂基础日粮)、肉鸡感染产气荚膜梭菌组(饲喂基础日粮)、感染产气荚膜梭菌肉鸡饲喂添加杆菌肽锌(40mg-kg-1)日粮、感染产气荚膜梭菌肉鸡饲喂添加5%沸石日粮和感染产气荚膜梭菌肉鸡饲喂添加5%沸石日粮及杆菌肽锌(40mg.kg-1),研究沸石对感染产气荚膜梭菌的肉鸡生产性能、免疫器官及屠宰性能的影响。结果表明：添加沸石显著提高了肉鸡前期平均体重和平均日增重(P<0.05),抗生素明显提高了肉鸡生产性能,但抗生素与沸石同时添加不影响肉鸡各阶段生产性能(P>0.05)。对免疫器官指数影响不明显(P>0.05)。感染产气荚膜梭菌使肉鸡血清唾液酸含量提高了21.64%(P>0.05),但各处理对感染产气荚膜梭菌肉鸡的血清唾液酸含量影响不明显(P>0.05)。添加抗生素、沸石、抗生素与沸石后则使肉鸡胸肌率和全净膛率均有不同程度的提高(P>0.05)。
7、天然沸石缓解饲料中低剂量铅对肉鸡毒性作用及相关机理研究
试验一：将720只1日龄AA肉鸡随机分成6组,分别为基础日粮对照组、基础日粮+3%沸石组、基础日粮+5%沸石组、基础日粮+硝酸铅(10mg.kg-1Pb)组、基础日粮+硝酸铅+3%沸石组、基础日粮+硝酸铅+5%沸石组,研究天然沸石和饲料中低剂量铅对肉鸡生产性能及血清HSP70的影响。结果表明：饲料中低剂量污染铅不影响肉鸡生产性能；添加3%沸石、低剂量污染铅并添加3%和5%的沸石则显著提高了肉鸡后期增重和降低了料重比(P<0.05),低剂量污染铅并添加5%的沸石使肉鸡全期平均日增重、饲料转化率均显著改善(P<0.05)。单独添加沸石和低剂量污染铅均不同程度的提高了肉鸡血清HSP70的(P>0.05),低剂量污染铅并添加3%和5%的沸石则显著提高了21日龄肉鸡血清HSP70水平(P<0.05),42日龄时肉鸡血清HSP70水平也有一定程度提高。
试验二：本试验设4个处理,分别为基础日粮对照组、基础日粮+硝酸铅(10mg-kg-1Pb)组、基础日粮十5%沸石、基础日粮+硝酸铅+5%沸石(10mg.kg-1Pb),研究天然沸石和饲料中低剂量铅对肉鸡抗体滴度、肝脏抗氧化功能与金属硫蛋白、胸腺细胞凋亡相关基因表达的影响。结果表明：16日龄时,沸石使肉鸡免疫抗体效价有所提高,37日龄时铅和沸石均显著提高了免疫抗体效价(P<0.05),其它则无明显差异。沸石可以减少铅在肉鸡肌肉中的残留,且沸石均极显著地降低了42日龄肉鸡肌肉中的铅残留(P<0.01),但各处理对肉鸡后期肝脏的抗氧化功能影响不大。污染铅显著提高了肉鸡21日龄时肝脏中的金属硫蛋白含量(P<0.05),添加沸石和污染铅再添加沸石对肉鸡肝脏中金属硫蛋白的含量影响不大(P>0.05)。污染铅显著提高了42日龄肉鸡胸腺bax基因的表达量(P<0.05)；污染铅并添加沸石有下调bax基因表达的趋势,并显著提高了21日龄时肉鸡胸腺bc1-2基因的表达量(P<0.05),各处理均显著提高了42日龄时bcl-2基因的表达。污染铅和添加沸石均显著降低了肉鸡21日龄时p53基因的表达量(P<0.05),污染铅使42日龄时的表达量明显提高(P<0.05),添加沸石则极显著地提高了42日龄时的p53基因表达量(P<0.01)。
The zeolites are functional materials with unique properties and wide uses, and have potentiality in the life science. This study was conducted to evaluate the quality and stability of natural zeolite (clinoptilolite) in vitro by chemical and X-ray diffraction. The effects of different quality and particle sizes, and modified natural zeolite on nutrition and other biological function in broilers were compared. The ameliorative effects of dietary natural zeolite (clinoptilolite) on performance, serum diamine oxidase (DAO) and nitric oxide (NO) level, apoptosis of thymocytes in broilers after Clostridium Perfringens infection were studied. The ameliorative effects of dietary natural zeolite (clinoptilolite) on performance, lead residual in breast muscle, OD value of serum HSP70, and gene expression of apoptosis cell in broilers after low dose lead exposure were researched. Then, the mechanism of harmful agent reduction by zeolite in broilers was explained.
1Evaluation of the quality and stability of natural zeolite (clinoptilolite)
Two zeolite samples were modified and treated under pH environment of broiler digestive tract and added NH4+, and then all samples were analyzed with X-ray diffraction, cationic exchange capacity and scanning electron microscopy method. The results showed that the composition of sample I mainly consisted of clinoptilolite above50%, and feldspar, hydromica, amphibole, silica. The chemical composition of sample I mainly contained Si and Al, and also contained Ca, Fe, K, Mg, and Na. The clinoptilolite content of sample Ⅱ was lower, and also occurred feldspar, montmorillonite. After the samples were treated under pH environment of broiler digestive tract, the clinoptilolite content of sample I decreased slightly, and Pb content decreased by39.2%and36.8%with or without NH4+, respectively. The treatments had no effect on the clinoptilolite content of sample Ⅱ, but sepiolite occurred. The cationic exchange capacity of zeolites increased by modification. The zeolites had good stability.
2Effects of different quality and modified zeolite on performance,digestibility of nutrients and biochemical indexes in blood of broilers
An experiment was conducted to study the effect of different quality and modified zeolites added in diets on the performance,the digestibility of nutrients and biochemical indexes in blood of broiler. A total of504one-day-old Arbor Acres broiler chickens were assigned randomly to four treatments with six replicates for each.The broilers were fed control diet and the same diet added with5%common zeolite, high quality zeolite and modified high quality zeolite, respectively. The results indicated that the supplementation of common zeolite, high quality zeolite and modified high quality zeolite in diet increased BWG of broiler by2.05%,7.62%(P<0.05) and4.73%(P<00.05),improved FCR by0.98%,5.39%(P<00.05) and3.43%(P<00.05) in finisher phase, respectively. The high quality zeolite and modified zeolite increased BWG of broiler by4.10%and2.41%,respectively, improved FCR by1.56%in whole growing phase. But the supplementation of all zeolites decreased the performance of broiler in starter phase (P<0.05).Compared with control group, the digestibility of dry matter, protein, energy of diets increased by4.05%,5.86%and3.03%;2.25%,6.16%and11.41%;3.59%,7.60%and7.16%with addition of three zeolites, but there was no effect on digestibility of calcium. The high quality zeolite and modified zeolite improved the digestibility of phosphorus. Three zeolites increased digestibility of all amino acid except methionine, the apparent digestibility of total essential amino acid increased, total non-essential amino acid increased and total amino acid by2.69%,4.79%and4.53%(P>0.05), by2.77%,7.56%and6.08%(P>0.05) and by2.74%,6.11%and5.26%(P>0.05) respectively. The modified high quality zeolite reduced the moisture of broiler feces. In conclusion, the supplementation of zeolites in diet improved the performance of broiler chicks and the digestibility of dietary nutrients, and the high quality zeolite was more effective. The addition of zeolites in diets had no effect on biochemical indexes in blood of broilers.
3Effects of different quality and modified zeolite on the enzyme activities, morphological structure and the DNA and RNA content of the small intestine for broilers
The experiment was conducted to analyze the effect of different quality and modified zeolites added in diets on the DNA and RNA content, activities of digestive enzymes, and morphological structure of the small intestine in broilers. The results indicated that, addition of all zeolites increased the RNA content in intestinal mucosa of broiler, and increased the DNA content of jejunum for finisher broiler(P<0.05).The common zeolite and high quality zeolite enhanced the DNA content of jejunum of starter broilers (P>0.05), but, all zeolites decreased the DNA content of ileum for starter broilers (P>0.05). The addition of three kinds of zeolites increased the amylase and protease activity in small intestine23.4%,8.3%,15.9%, and17.4%,17.2%,2.9%at starter phase (P>0.05),and protease activity increased by4.0%,25.2%,2.8%(P>0.05) at grower phase for broilers. The common zeolite increased the amylase activity by12.8%at grower phase,but high quality zeolite and modified quality zeolite decreased the amylase activity by25.1%and14.1%(P>0.05).The zeolites improved the morphological structure of the small intestine in broilers.
4Effects of different particle sizes zeolites on the performance, intestinal microflora and residual of heavy metal and effect of zeolite added into different form of feed on performance in broilers
This experiment was conducted to study the effects of zeolites of three different particle sizes added to diets on the performance, intestinal microflora and residual of heavy metal in broilers. The results indicated that the supplementation of5%zeolite of coarse, middle and fine size to diets had no effect on BWG for broilers,but improved FCR (P<0.05), decreased the mortality of broilers. The diets added with coarse, middle and fine size zeolite decreased the Pb concentration of breast muscle by10.06%,43.02%and39.11%, and reduced the As content of liver by34.00%,29.85%and16.12%, respectively, but the As content of breast muscle was unaffected. The coarse and middle size zeolite decreased the Pb concentration of liver by32.61%and24.78%. Blood ammonia level and the number of E.coil in jejunum were decreased markedly with zeolite treatment, and the number of Lactobacillus in jejunum was increased.
The performance of broilers fed with pellet feed was better than that with mash form (P<0.05). The addition of zeolite in pellet feed had no effect on performance of broilers at starter phase, but addition of zeolite in mash feed reduced growth, feed consumption, and FCR (P<0.05). The pellet feed added with zeolite decreased growth and FCR, but no effect on feed intake. The zeolite in mash feed had no effect on growth at finisher phase, but reduced feed intake and FCR (P<0.05). The zeolite added in pellet or mash feed had no effect on gain, but dencreased FCR for pellet (P<0.05),and increased for mash during full growth period.
5The alleviative effects of dietary natural zeolite (clinoptilolite) on broilers after Clostridium Perfringens infection
3841-day-old Arbor Acres chicks were randomly assigned to4treatment groups consisting of6replicates to study the alleviative effect of natural zeolite on broilers after Clostridium Perfringens infection. The details of four treatments were described as follows: Control(broilers fed with basal diet), Clostridium Perfringens infection broilers fed with basal diet, Clostridium Perfringens infection broilers fed with basal diet added zinc bacitracin at40mg·kg-1, and Clostridium Perfringens infection broilers fed with basal diet added5%natural zeolite. The results showed that the natural zeolite significantly increased FCR(P<0.05) at starter phase, but all treatments had no significant effect on organ index and serum biochemical parameters of broilers by Clostridium Perfringens infection. Compared to the infection treatment, the zeolite increased the activity of T-SOD level in serum at starter phase (P<0.01). The result documented that the natural zeolite alleviated the decline of antioxidation by Clostridium Perfringens infection in broilers. The addition of zinc bacitracin in diet had the same effect. The Clostridium Perfringens infection increased serum NO level at starter phase (P<0.01) and the activity of DAO at grower phase for broilers. The zeolite decreased the serum NO production at starter phase (P<0.01) and the activity of DAO in serum at grower phase(P<0.05), and the result implied the natural zeolite alleviated the entero-lesion of broilers by Clostridium Perfringens. The zeolite and zinc bacitracin had no significant effect on T3, T4, IgA, and IgG in serum of broilers (P>0.05). The supplementation of natural zeolite in diet decreased the dead cells and increased the living cell of thymus in broilers after Clostridium Perfringen infection
6The effects of dietary natural zeolite (clinoptilolite) on performance, immune organ and carcass trait after Clostridium Perfringens infection in broilers
This trial was conducted to study the effects of natural zeolite on performance, immune organ and carcass trait of broilers after Clostridium Perfringens infection. There were5treatments consisted of Control(broilers fed with basal diet), Clostridium Perfringens infection broilers fed with basal diet, Clostridium Perfringens infection broilers fed with basal diet added zinc bacitracin at40mg·-kg-1, Clostridium Perfringens infection broilers fed with basal diet added5%natural zeolite, and Clostridium Perfringens infection broilers fed with basal diet added zinc bacitracin at40mg·kg-1and5%natural zeolite. The results showed that natural zeolite significantly increased the body weight and ADG (P<0.05), and zinc bacitracin improved all parameters of performance of broilers at starter phase, but the combining addition of natural zeolite with zinc bacitracin in diet had no effect on the performance of broilers(P>0.05). All treatments had no significant effect on organ index of broilers. The content of SA in serum increased by21.64%after Clostridium Perfringens infection, but natural zeolite or/and zinc bacitracin in diet had no effect on the content of SA in serum. The supplementation of natural zeolite or/and zinc bacitracin slightly increased breast meat rate and dressing yield.
7The alleviative effects and mechanism of dietary natural zeolite (clinoptilolite) on broilers after low dose lead exposure
Trial1720I-day-old AA broilers were randomly assigned to six treatment groups consisting of8replicates to study effects of natural zeolite and low dose lead added in diet on performance and serum HSP70of broilers. The treatments were:control (broilers fed with basal diet), basal diet added with3%natural zeolite,5%natural zeolite,10mg·kg-1Pb,3%natural zeolite with10mg·kg-1Pb,5%natural zeolite with10mg·kg-1Pb, respectively. The results showed that low dose lead had no effect on performance of broilers. The addition of3%natural zeolite, the low dose lead with3%and5zeolite increased BWG and decreased FCR at grower phase (P<0.05), and the addition of5%natural zeolite and lead of10mg·kg-1increased BWG and FCR (P<0.05) in whole period. The supplementation of natural zeolite or low dose lead in diet increased the serum HSP70OD value (P>0.05), the low dose lead with3%or5%natural zeolite increased HSP70level at grower phase (P<0.05)
Trial2This experiment was carried out to study alleviative effects of natural zeolite on antibody titer、liver antioxidation function and expression of apoptosis associated genes of broilers during broiler fed diet with low dose lead addition. The treatments included: control (broilers fed with basal diet), basal diet added with10mg·kg-1Pb,5%natural zeolite,10mg·kg-1Pb with5%natural zeolite, respectively. The results showed that natural zeolite increased antibody titer of ND for broilers at16d and at37d (P<0.05), the supplementation of low dose lead did too(P<0.05). The natural zeolite reduced the residual of lead in breast of broilers, especially at42d (P<0.01).But all treatments had no significant effect on antioxidation function of broiler liver. The MT content of liver was increased in starter phase of chickens fed diet with low dose lead addition(P<0.05), but natural zeolite with or without low dose lead had no effect on the content of MT (P>0.05) The low dose lead increased the expression level of bax gene at42d (P<0.05), the zeolite decreased slightly bax gene level which result in increasing by lead (P>0.05), and significantly increased the expression level of bcl-2gene in thymus of broilers at21d. All treatments significantly enhanced bcl-2level. The supplementation of low dose lead or natural zeolite decreased the expression level of p53gene at21d (P<0.05),the low dose lead (P<0.05) or natural zeolite (P<0.01) in diet enhanced p53level of broilers at42d.
1、天然沸石的品位及稳定性评价
采集两种天然沸石样品,进行改性处理和模拟消化道pH环境以及加NH4+处理,采用X射线衍射、阳离子交换容量(吸铵值)和电镜扫描进行测定分析。结果表明：样品I的主成分为斜发沸石,含量达50%以上,其杂质主要是长石和水云母,并含有部分闪石和二氧化硅,化学成分主要是Si和Al,并含有一定量的Ca、Fe、K、Mg、Na;样品Ⅱ的斜发沸石含量较低,杂质主要是长石和蒙脱石。经模拟消化道环境处理后,样品I斜发沸石的含量稍有下降,Pb含量分别下降了39.2%和36.8%；样品II斜发沸石含量未改变,但出现了海泡石。改性提高了沸石的阳离子交换容量；结果提示试验所用沸石具有良好的稳定性。
2、不同品位及改性沸石对肉鸡生产性能、养分表观代谢率、血液生化指标的影响
将504只1日龄AA肉鸡随机分为四个处理,每个处理6个重复,分别饲喂对照日粮,对照日粮添加5%普通沸石、高品位沸石和高品位改性沸石的试验日粮,研究不同品位沸石及改性沸石对肉鸡生产性能、养分表观消化率、血液生化指标的影响。结果表明：添加沸石后肉鸡后期的体增重分别比对照组提高了2.05%、7.62%(P<0.05)和4.73%(P<0.05),饲料转化率改善了0.98%、5.39%(P<0.05)和3.43%(P<0.05)；高品位和改性高品位沸石使肉鸡全期的体重提高了4.10%和2.41%,饲料转化率均改善1.56%；但添加沸石降低了肉鸡前期体重,而提高了饲料转化率(P<0.05)。与对照组相比,三种沸石使饲料干物质的表观消化率分别提高了4.05%、5.86%及3.03%,粗蛋白消化率分别提高了2.25%、6.16%和11.410%,能量消化率分别提高了3.59%、7.60%和7.16%；但未能提高钙的消化率,高品位和改性高品位沸石提高了磷的消化率；除蛋氨酸外,添加沸石提高了其他氨基酸的消化率,分别使总必需氨基酸表观消化率提高了2.69%、4.79%和4.53%(P>0.05),总非必需氨基酸表观消化率提高了2.77%、7.56%和6.080%(P>0.05),总氨基酸表观消化率提高了2.74%、6.1l%和5.26%(P>0.05)；改性高品位沸石降低了肉鸡粪便中的水分含量；沸石对肉鸡血液生化指标无明显影响。
3、不同品位及改性沸石对肉鸡肠道消化酶活性、DNA与RNA含量及黏膜形态的影响
就普通沸石、高品位沸石和高品位改性沸石对肉鸡肠道粘膜中的RNA及DNA含量、消化酶活性和形态结构的影响进行了测定分析。结果表明,日粮中分别添加5%普通沸石、高品位沸石和高品位改性沸石,可提高肉鸡前期和后期肠道粘膜中RNA含量及后期空肠粘膜中的DNA含量；普通和高品位沸石使肉鸡前期空肠粘膜中的DNA含量有所提高(P>0.05),但均降低了前期回肠粘膜中的DNA含量(P>0.05);对肉鸡后期肠道粘膜中的DNA含量则影响不一。饲料中添加三种沸石分别使肉鸡前期肠道淀粉酶和蛋白酶活性提高了23.4%、8.3%、15.9%和17.4%、17.2%、2.90%,后期蛋白酶活性提高了4.0%、25.2%、22.8%(P>0.05),普通沸石使肉鸡后期内源淀粉酶活性提高了12.8%,但高品位沸石和改性沸石却使其分别下降了25.1%和14.1%(P>0.05)。结果提示,沸石具有改善肉鸡肠道粘膜形态的功能。
4、不同粒度沸石对肉鸡生产性能、肠道菌群及组织重金属残留的影响以及沸石在不同料型饲料中的应用效果
在饲料中分别添加三种不同粒度的沸石粉,研究其对肉鸡生产性能、肠道菌群及肝脏与肌肉中重金属残留量的影响及沸石在不同料型饲料中的应用效果。结果表明,与对照组相比,在饲料中添加5%三种不同粒度的沸石均未影响肉鸡增重,但改善了饲料转化率(P<0.05),降低了死淘率；粗、中、细三种粒度沸石使肉鸡肌肉中的铅含量分别降低了10.06%、43.02%、39.11%,肝脏中砷含量分别下降了34.00%、29.85%、16.12%,粗、中粒度沸石使肝脏中铅含量分别降低了32.61%、24.78%,沸石对肌肉中的砷含量影响不大；沸石降低了血氨浓度及空肠中的大肠杆菌数,使空肠中的乳酸菌数有所提高。
饲喂颗粒饲料的肉鸡生产性能明显优于粉料(P<0.05)。颗粒饲料中添加沸石对肉鸡前期生产性能无影响,但在粉料中添加则显著降低了肉鸡生长速度、采食量和饲料转化率(P<0.05)。后期在颗粒饲料中添加沸石则降低了肉鸡生长速度(P<0.05)和饲料转化率(P>0.05),但不影响采食量；粉料后期对增重无影响,但明显降低了采食量和料重比(P<0.05)。颗粒料和粉料中添加沸石均不影响肉鸡全期增重,但颗粒料的饲料转化率有所上升(P>0.05),而粉料则明显降低(P<0.05)。
5、天然沸石缓解产气荚膜梭茵对肉鸡的有害影响及相关机制研究
384只1日龄的AA肉鸡随机分为4个处理,每个处理6个重复,分别为对照组(饲喂基础日粮)、肉鸡感染产气荚膜梭菌组(饲喂基础日粮)、感染产气荚膜梭菌肉鸡饲喂添加杆菌肽锌(40mg.kg-1)日粮和感染产气荚膜梭菌肉鸡饲喂添加5%沸石日粮,研究天然沸石缓解产气荚膜梭菌对肉鸡的有害影响及相关机制。结果表明：沸石明显提高了感染产气荚膜组肉鸡前期的饲料转化率(P<0.05),但对免疫器官指数和血清各生化指标均影响不明显(P>0.05)。与感染组相比,饲料中添加沸石使肉鸡前期血清SOD极显著提高(P<0.01),沸石缓解了产气荚膜梭菌对肉鸡造成的氧化损伤；抗生素也有相同效果。产气荚膜梭菌感染明显提高了肉鸡前期NO和后期DAO;沸石显著降低了肉鸡前期血清NO(P<0.01)和后期DAO(P<0.05),沸石可缓解产气荚膜梭菌对肠道的损伤；抗生素亦具同样作用。沸石和抗生素对肉鸡血清T3、T4、IgA和IgG均无影响(P>0.05)；但添加沸石可减少产气荚膜梭菌导致的胸腺细胞死亡数,提高活细胞数量(P>0.05)。
6、沸石对感染产气荚膜梭菌肉鸡生产性能、免疫器官及屠宰性能的影响
本试验设5个处理,分别为对照组(饲喂基础日粮)、肉鸡感染产气荚膜梭菌组(饲喂基础日粮)、感染产气荚膜梭菌肉鸡饲喂添加杆菌肽锌(40mg-kg-1)日粮、感染产气荚膜梭菌肉鸡饲喂添加5%沸石日粮和感染产气荚膜梭菌肉鸡饲喂添加5%沸石日粮及杆菌肽锌(40mg.kg-1),研究沸石对感染产气荚膜梭菌的肉鸡生产性能、免疫器官及屠宰性能的影响。结果表明：添加沸石显著提高了肉鸡前期平均体重和平均日增重(P<0.05),抗生素明显提高了肉鸡生产性能,但抗生素与沸石同时添加不影响肉鸡各阶段生产性能(P>0.05)。对免疫器官指数影响不明显(P>0.05)。感染产气荚膜梭菌使肉鸡血清唾液酸含量提高了21.64%(P>0.05),但各处理对感染产气荚膜梭菌肉鸡的血清唾液酸含量影响不明显(P>0.05)。添加抗生素、沸石、抗生素与沸石后则使肉鸡胸肌率和全净膛率均有不同程度的提高(P>0.05)。
7、天然沸石缓解饲料中低剂量铅对肉鸡毒性作用及相关机理研究
试验一：将720只1日龄AA肉鸡随机分成6组,分别为基础日粮对照组、基础日粮+3%沸石组、基础日粮+5%沸石组、基础日粮+硝酸铅(10mg.kg-1Pb)组、基础日粮+硝酸铅+3%沸石组、基础日粮+硝酸铅+5%沸石组,研究天然沸石和饲料中低剂量铅对肉鸡生产性能及血清HSP70的影响。结果表明：饲料中低剂量污染铅不影响肉鸡生产性能；添加3%沸石、低剂量污染铅并添加3%和5%的沸石则显著提高了肉鸡后期增重和降低了料重比(P<0.05),低剂量污染铅并添加5%的沸石使肉鸡全期平均日增重、饲料转化率均显著改善(P<0.05)。单独添加沸石和低剂量污染铅均不同程度的提高了肉鸡血清HSP70的(P>0.05),低剂量污染铅并添加3%和5%的沸石则显著提高了21日龄肉鸡血清HSP70水平(P<0.05),42日龄时肉鸡血清HSP70水平也有一定程度提高。
试验二：本试验设4个处理,分别为基础日粮对照组、基础日粮+硝酸铅(10mg-kg-1Pb)组、基础日粮十5%沸石、基础日粮+硝酸铅+5%沸石(10mg.kg-1Pb),研究天然沸石和饲料中低剂量铅对肉鸡抗体滴度、肝脏抗氧化功能与金属硫蛋白、胸腺细胞凋亡相关基因表达的影响。结果表明：16日龄时,沸石使肉鸡免疫抗体效价有所提高,37日龄时铅和沸石均显著提高了免疫抗体效价(P<0.05),其它则无明显差异。沸石可以减少铅在肉鸡肌肉中的残留,且沸石均极显著地降低了42日龄肉鸡肌肉中的铅残留(P<0.01),但各处理对肉鸡后期肝脏的抗氧化功能影响不大。污染铅显著提高了肉鸡21日龄时肝脏中的金属硫蛋白含量(P<0.05),添加沸石和污染铅再添加沸石对肉鸡肝脏中金属硫蛋白的含量影响不大(P>0.05)。污染铅显著提高了42日龄肉鸡胸腺bax基因的表达量(P<0.05)；污染铅并添加沸石有下调bax基因表达的趋势,并显著提高了21日龄时肉鸡胸腺bc1-2基因的表达量(P<0.05),各处理均显著提高了42日龄时bcl-2基因的表达。污染铅和添加沸石均显著降低了肉鸡21日龄时p53基因的表达量(P<0.05),污染铅使42日龄时的表达量明显提高(P<0.05),添加沸石则极显著地提高了42日龄时的p53基因表达量(P<0.01)。
The zeolites are functional materials with unique properties and wide uses, and have potentiality in the life science. This study was conducted to evaluate the quality and stability of natural zeolite (clinoptilolite) in vitro by chemical and X-ray diffraction. The effects of different quality and particle sizes, and modified natural zeolite on nutrition and other biological function in broilers were compared. The ameliorative effects of dietary natural zeolite (clinoptilolite) on performance, serum diamine oxidase (DAO) and nitric oxide (NO) level, apoptosis of thymocytes in broilers after Clostridium Perfringens infection were studied. The ameliorative effects of dietary natural zeolite (clinoptilolite) on performance, lead residual in breast muscle, OD value of serum HSP70, and gene expression of apoptosis cell in broilers after low dose lead exposure were researched. Then, the mechanism of harmful agent reduction by zeolite in broilers was explained.
1Evaluation of the quality and stability of natural zeolite (clinoptilolite)
Two zeolite samples were modified and treated under pH environment of broiler digestive tract and added NH4+, and then all samples were analyzed with X-ray diffraction, cationic exchange capacity and scanning electron microscopy method. The results showed that the composition of sample I mainly consisted of clinoptilolite above50%, and feldspar, hydromica, amphibole, silica. The chemical composition of sample I mainly contained Si and Al, and also contained Ca, Fe, K, Mg, and Na. The clinoptilolite content of sample Ⅱ was lower, and also occurred feldspar, montmorillonite. After the samples were treated under pH environment of broiler digestive tract, the clinoptilolite content of sample I decreased slightly, and Pb content decreased by39.2%and36.8%with or without NH4+, respectively. The treatments had no effect on the clinoptilolite content of sample Ⅱ, but sepiolite occurred. The cationic exchange capacity of zeolites increased by modification. The zeolites had good stability.
2Effects of different quality and modified zeolite on performance,digestibility of nutrients and biochemical indexes in blood of broilers
An experiment was conducted to study the effect of different quality and modified zeolites added in diets on the performance,the digestibility of nutrients and biochemical indexes in blood of broiler. A total of504one-day-old Arbor Acres broiler chickens were assigned randomly to four treatments with six replicates for each.The broilers were fed control diet and the same diet added with5%common zeolite, high quality zeolite and modified high quality zeolite, respectively. The results indicated that the supplementation of common zeolite, high quality zeolite and modified high quality zeolite in diet increased BWG of broiler by2.05%,7.62%(P<0.05) and4.73%(P<00.05),improved FCR by0.98%,5.39%(P<00.05) and3.43%(P<00.05) in finisher phase, respectively. The high quality zeolite and modified zeolite increased BWG of broiler by4.10%and2.41%,respectively, improved FCR by1.56%in whole growing phase. But the supplementation of all zeolites decreased the performance of broiler in starter phase (P<0.05).Compared with control group, the digestibility of dry matter, protein, energy of diets increased by4.05%,5.86%and3.03%;2.25%,6.16%and11.41%;3.59%,7.60%and7.16%with addition of three zeolites, but there was no effect on digestibility of calcium. The high quality zeolite and modified zeolite improved the digestibility of phosphorus. Three zeolites increased digestibility of all amino acid except methionine, the apparent digestibility of total essential amino acid increased, total non-essential amino acid increased and total amino acid by2.69%,4.79%and4.53%(P>0.05), by2.77%,7.56%and6.08%(P>0.05) and by2.74%,6.11%and5.26%(P>0.05) respectively. The modified high quality zeolite reduced the moisture of broiler feces. In conclusion, the supplementation of zeolites in diet improved the performance of broiler chicks and the digestibility of dietary nutrients, and the high quality zeolite was more effective. The addition of zeolites in diets had no effect on biochemical indexes in blood of broilers.
3Effects of different quality and modified zeolite on the enzyme activities, morphological structure and the DNA and RNA content of the small intestine for broilers
The experiment was conducted to analyze the effect of different quality and modified zeolites added in diets on the DNA and RNA content, activities of digestive enzymes, and morphological structure of the small intestine in broilers. The results indicated that, addition of all zeolites increased the RNA content in intestinal mucosa of broiler, and increased the DNA content of jejunum for finisher broiler(P<0.05).The common zeolite and high quality zeolite enhanced the DNA content of jejunum of starter broilers (P>0.05), but, all zeolites decreased the DNA content of ileum for starter broilers (P>0.05). The addition of three kinds of zeolites increased the amylase and protease activity in small intestine23.4%,8.3%,15.9%, and17.4%,17.2%,2.9%at starter phase (P>0.05),and protease activity increased by4.0%,25.2%,2.8%(P>0.05) at grower phase for broilers. The common zeolite increased the amylase activity by12.8%at grower phase,but high quality zeolite and modified quality zeolite decreased the amylase activity by25.1%and14.1%(P>0.05).The zeolites improved the morphological structure of the small intestine in broilers.
4Effects of different particle sizes zeolites on the performance, intestinal microflora and residual of heavy metal and effect of zeolite added into different form of feed on performance in broilers
This experiment was conducted to study the effects of zeolites of three different particle sizes added to diets on the performance, intestinal microflora and residual of heavy metal in broilers. The results indicated that the supplementation of5%zeolite of coarse, middle and fine size to diets had no effect on BWG for broilers,but improved FCR (P<0.05), decreased the mortality of broilers. The diets added with coarse, middle and fine size zeolite decreased the Pb concentration of breast muscle by10.06%,43.02%and39.11%, and reduced the As content of liver by34.00%,29.85%and16.12%, respectively, but the As content of breast muscle was unaffected. The coarse and middle size zeolite decreased the Pb concentration of liver by32.61%and24.78%. Blood ammonia level and the number of E.coil in jejunum were decreased markedly with zeolite treatment, and the number of Lactobacillus in jejunum was increased.
The performance of broilers fed with pellet feed was better than that with mash form (P<0.05). The addition of zeolite in pellet feed had no effect on performance of broilers at starter phase, but addition of zeolite in mash feed reduced growth, feed consumption, and FCR (P<0.05). The pellet feed added with zeolite decreased growth and FCR, but no effect on feed intake. The zeolite in mash feed had no effect on growth at finisher phase, but reduced feed intake and FCR (P<0.05). The zeolite added in pellet or mash feed had no effect on gain, but dencreased FCR for pellet (P<0.05),and increased for mash during full growth period.
5The alleviative effects of dietary natural zeolite (clinoptilolite) on broilers after Clostridium Perfringens infection
3841-day-old Arbor Acres chicks were randomly assigned to4treatment groups consisting of6replicates to study the alleviative effect of natural zeolite on broilers after Clostridium Perfringens infection. The details of four treatments were described as follows: Control(broilers fed with basal diet), Clostridium Perfringens infection broilers fed with basal diet, Clostridium Perfringens infection broilers fed with basal diet added zinc bacitracin at40mg·kg-1, and Clostridium Perfringens infection broilers fed with basal diet added5%natural zeolite. The results showed that the natural zeolite significantly increased FCR(P<0.05) at starter phase, but all treatments had no significant effect on organ index and serum biochemical parameters of broilers by Clostridium Perfringens infection. Compared to the infection treatment, the zeolite increased the activity of T-SOD level in serum at starter phase (P<0.01). The result documented that the natural zeolite alleviated the decline of antioxidation by Clostridium Perfringens infection in broilers. The addition of zinc bacitracin in diet had the same effect. The Clostridium Perfringens infection increased serum NO level at starter phase (P<0.01) and the activity of DAO at grower phase for broilers. The zeolite decreased the serum NO production at starter phase (P<0.01) and the activity of DAO in serum at grower phase(P<0.05), and the result implied the natural zeolite alleviated the entero-lesion of broilers by Clostridium Perfringens. The zeolite and zinc bacitracin had no significant effect on T3, T4, IgA, and IgG in serum of broilers (P>0.05). The supplementation of natural zeolite in diet decreased the dead cells and increased the living cell of thymus in broilers after Clostridium Perfringen infection
6The effects of dietary natural zeolite (clinoptilolite) on performance, immune organ and carcass trait after Clostridium Perfringens infection in broilers
This trial was conducted to study the effects of natural zeolite on performance, immune organ and carcass trait of broilers after Clostridium Perfringens infection. There were5treatments consisted of Control(broilers fed with basal diet), Clostridium Perfringens infection broilers fed with basal diet, Clostridium Perfringens infection broilers fed with basal diet added zinc bacitracin at40mg·-kg-1, Clostridium Perfringens infection broilers fed with basal diet added5%natural zeolite, and Clostridium Perfringens infection broilers fed with basal diet added zinc bacitracin at40mg·kg-1and5%natural zeolite. The results showed that natural zeolite significantly increased the body weight and ADG (P<0.05), and zinc bacitracin improved all parameters of performance of broilers at starter phase, but the combining addition of natural zeolite with zinc bacitracin in diet had no effect on the performance of broilers(P>0.05). All treatments had no significant effect on organ index of broilers. The content of SA in serum increased by21.64%after Clostridium Perfringens infection, but natural zeolite or/and zinc bacitracin in diet had no effect on the content of SA in serum. The supplementation of natural zeolite or/and zinc bacitracin slightly increased breast meat rate and dressing yield.
7The alleviative effects and mechanism of dietary natural zeolite (clinoptilolite) on broilers after low dose lead exposure
Trial1720I-day-old AA broilers were randomly assigned to six treatment groups consisting of8replicates to study effects of natural zeolite and low dose lead added in diet on performance and serum HSP70of broilers. The treatments were:control (broilers fed with basal diet), basal diet added with3%natural zeolite,5%natural zeolite,10mg·kg-1Pb,3%natural zeolite with10mg·kg-1Pb,5%natural zeolite with10mg·kg-1Pb, respectively. The results showed that low dose lead had no effect on performance of broilers. The addition of3%natural zeolite, the low dose lead with3%and5zeolite increased BWG and decreased FCR at grower phase (P<0.05), and the addition of5%natural zeolite and lead of10mg·kg-1increased BWG and FCR (P<0.05) in whole period. The supplementation of natural zeolite or low dose lead in diet increased the serum HSP70OD value (P>0.05), the low dose lead with3%or5%natural zeolite increased HSP70level at grower phase (P<0.05)
Trial2This experiment was carried out to study alleviative effects of natural zeolite on antibody titer、liver antioxidation function and expression of apoptosis associated genes of broilers during broiler fed diet with low dose lead addition. The treatments included: control (broilers fed with basal diet), basal diet added with10mg·kg-1Pb,5%natural zeolite,10mg·kg-1Pb with5%natural zeolite, respectively. The results showed that natural zeolite increased antibody titer of ND for broilers at16d and at37d (P<0.05), the supplementation of low dose lead did too(P<0.05). The natural zeolite reduced the residual of lead in breast of broilers, especially at42d (P<0.01).But all treatments had no significant effect on antioxidation function of broiler liver. The MT content of liver was increased in starter phase of chickens fed diet with low dose lead addition(P<0.05), but natural zeolite with or without low dose lead had no effect on the content of MT (P>0.05) The low dose lead increased the expression level of bax gene at42d (P<0.05), the zeolite decreased slightly bax gene level which result in increasing by lead (P>0.05), and significantly increased the expression level of bcl-2gene in thymus of broilers at21d. All treatments significantly enhanced bcl-2level. The supplementation of low dose lead or natural zeolite decreased the expression level of p53gene at21d (P<0.05),the low dose lead (P<0.05) or natural zeolite (P<0.01) in diet enhanced p53level of broilers at42d.
引文
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