自然霉变玉米对肉鸭生产性能和消化生理的影响及机制研究
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摘要
饲料污染霉菌毒素会降低动物生产性能,抑制免疫机能,甚至在动物组织和产品中残留,给养殖业造成了巨大危害。近年的研究表明,霉菌毒素可改变肠道的消化功能。肉鸭对霉菌毒素中的黄曲霉毒素B1 (AFB1)比其他动物更敏感。本研究通过考察自然霉变玉米对肉鸭生产性能、消化器官发育、肠道形态结构、消化酶活性、各肠道DNA和RNA含量、营养物质消化率、食糜通过速度、Na+,K+-ATPase基因表达量和内分泌激素的影响来探讨霉菌毒素对肉鸭消化生理和消化功能的影响及其可能机制。研究共包括四个试验。
试验一自然霉变玉米对肉鸭生产性能和肠道形态结构的影响
本试验旨在研究以自然霉变玉米配制饲粮对肉鸭生产性能和肠道形态结构的影响。试验设2个处理,处理1为正常玉米对照组,处理2以霉变玉米(AFB1超标)100%替代正常玉米。每个处理6个重复,每个重复16只1日龄健康樱桃谷肉鸭(公母各半)。试验期4w。在试验的第0d、14d和28d按重复称肉鸭体重,并计算耗料；试验第28d每重复选2只肉鸭屠宰,采集空肠组织观测肠道形态结构。试验结果显示,自然霉变玉米组死亡率极显著升高(p<0.01),生产性能显著下降：与对照组相比,0-28d日增重和日采食量分别降低了19.96%(p<0.01)和18.50%(p<0.05),0-28d料重比略低(p>0.05)而0-14d料重比极显著降低(p<0.01)；自然霉变玉米组肉鸭肠绒毛高度较正常玉米组提高了22.11%(p<0.05),隐窝深度降低了32.69%(p<0.05),绒毛高度/隐窝深度极显著提高(p<0.01),肠壁厚略有提高(p>0.05)；自然霉变玉米组肉鸭发生心包积水、肝脏褐色素沉积及肝组织灶性坏死、结缔组织增生,淋巴细胞和脂肪细胞增多等病变。结果表明：摄食含AFB1超标自然霉变玉米的饲粮将导致肉鸭死亡率升高和生产性能降低,导致肝脏等内脏发生病变,并明显影响肠道形态结构。
试验二自然霉变玉米对肉鸭生产性能、肠道发育及消化酶活性的影响
本试验旨在研究肉鸭摄食以污染多种霉菌毒素且AFB1超标的自然霉变玉米配制的饲粮后,生产性能、肠道发育和消化酶活性的变化。试验采用单因子设计,共3个处理组,即用自然霉变玉米分别按0%(对照组)、50%和100%比例替代饲粮中正常玉米,每个处理8个重复,每个重复16只肉鸭,公母各半。试验期5w。试验考察了肉鸭各阶段的生产性能,观测了第14 d和35 d的消化器官指数和肠道形态,测定了第14 d的肠黏膜RNA和蛋白质含量,以及第14 d的空肠刷状缘酶与胰腺酶活性。结果显示：在整个试验期内,随着霉变玉米在饲粮中用量增加,肉鸭的ADFI、ADG、F/G均呈线性降低(p<0.001),死亡率线性升高(p<0.001),100%霉变玉米组鸭的死亡率达31.25%;第14d和35d的十二指肠指数、空肠指数、回肠指数及第35d的肝脏指数均较其余两组极显著增加(p<0.001)；随霉变玉米的用量增加,第14d空肠粘膜的绒毛宽度(VW)(p<0.05)、绒毛高度(VH)(p<0.001)和绒毛表面积(VASA)(p<0.01)增加,第35d十二指肠粘膜的隐窝深度(CD)(p<0.01).VW(p=0.10).VH(p<0.05)和VASA(p<0.05)增加,第14d十二指肠和空肠粘膜的DNA含量增加而RNA含量降低,第14d空肠刷状缘碱性磷酸酶活性(p<0.05)和亮氨酸氨肽酶活性(p<0.05)以及胰腺胰蛋白酶活性(p<0.05)和糜蛋白酶活性(p<0.001)升高。结果表明：自然霉变玉米饲粮(AFB1超标)使肉鸭采食受到抑制,增重减慢,死亡率增加,改变肉鸭肠道发育和消化酶活性。
试验三自然霉变玉米对肉鸭食糜通过速率和养分消化率的影响
本试验旨在研究以自然霉变玉米配制的饲粮对肉鸭食糜通过速率和养分消化率的影响。试验设3个处理,处理1为正常玉米组,处理2为霉变玉米组,处理3为采食配对组,即肉鸭采食的饲粮同处理1而采食的量同处理2,每个处理8个重复,每个重复16只肉鸭。试验期14d。试验考察了肉鸭的生产性能、养分消化率和食糜流通速率。结果显示：在0-7d、8-14d和0-14d,与正常玉米组相比,自然霉变玉米组肉鸭ADFI分别降低了23.55%(p<0.01)、47.56%(p<0.01)和43.23%(p<0.01),ADG分别降低了31.47%(p<0.01)、41.57%(p<0.01)和38.84%(p<0.01),与采食配对组相比,自然霉变玉米组ADG分别增加4.63%、18.25%和13.83%(p<0.01)；与正常玉米组和采食配对组相比,霉变玉米组全期F/G分别降低4.90%(p<0.01)和5.56%(p<0.01),DM消化率分别提高3.78%(p<0.01)和3.75%(p<0.01),CP消化率分别提高2.65%和4.63%(p<0.01),总氨基酸消化率分别提高3.9%(p<0.01)和3.98%(p<0.01)。同时,自然霉变玉米组食糜在消化道各段的停留时间延长了,与正常玉米组相比,食管膨大部和空肠停留时间分别延长37.78%(p<0.05)和55.94%(p<0.05)。结果表明：以自然霉变玉米配制饲粮(AFB1超标)饲喂肉鸭,会抑制肉鸭采食,降低肉鸭增重,同时延长食糜在消化道的停留时间,提高养分消化率、降低料重比。
试验四自然霉变玉米对肉鸭血清激素水平和肠组织Na+,K+-ATPase基因表达的影响
为了进一步探明自然霉变玉米影响肉鸭摄食及消化生理的机制,本试验研究了以自然霉变玉米配制的饲粮对肉鸭血清采食调控相关激素水平和肠组织Na+,K+-ATPase基因表达的影响。试验设2个处理,处理1为正常玉米饲粮组,处理2饲粮则以自然霉变玉米(AFB1超标)100%替代正常玉米配制,每个处理8个重复,每个重复16只1日龄健康肉鸭。试验期28天。第14 d和第28 d,分别从每个重复中选取2只肉鸭(公母各一只),颈静脉采血4ml,测定血清中神经肽Y (NPY)、Ghrelin、瘦素(Leptin)、八肽胆囊收缩素(CCK-8)、胰岛素、胰岛素样生长因子-1(IGF-1)的含量。第14 d采血后处死肉鸭,采集空肠前段(前1/4处)组织,检测Na+,K+-ATPase基因相对表达量。结果显示：自然霉变玉米组肉鸭14d和28d的血清中IGF-1和Leptin的含量皆极显著提高(p<0.01),而第28d血清NPY含量降低(p<0.05)；肉鸭空肠组织Na+,K+-ATPase基因相对表达量较对照组增加了23%(p>0.05)。结果表明：肉鸭采食自然霉变玉米饲粮(AFB1超标)后,可能通过提高血清Leptin并降低血清NPY含量,进而抑制肉鸭采食；可能通过提高血清IGF-1含量和肠组织Na+, K+-ATPase基因表达量来影响养分吸收和饲料效率,缓解霉菌毒素的危害。
综上所述,以自然霉变玉米配制的饲粮饲喂肉鸭会导致死淘亡率升高,采食量、增重和料重比降低,饲料养分表观消化率提高。进一步分析发现,自然霉变玉米导致的增重下降主要是通过降低肉鸭的采食量所致,其机制与血清Leptin含量提高而NPY含量降低有关；而料重比较低的原因是饲料养分消化利用率提高,其机制与十二指肠和空肠粘膜的DNA含量增加、RNA含量降低,绒毛高度、宽度和表面积增加,空肠组织Na+, K+-ATPase基因表达量上升,空肠刷状缘的碱性磷酸酶和亮氨酸氨肽酶活性提高,胰腺胰蛋白酶和糜蛋白酶活性增强,以及食糜在消化道的通过速率减慢等因素的综合作用有关。结果提示,摄食含AFB1超标自然霉变玉米的饲粮对肉鸭消化系统和内分泌系统产生较复杂的影响,进而影响生产性能。
Feed contaminated with mycotoxins is harmful to animals. It reduces growth and depresses the immunity of animals. These effects have serious economic implications for animal producers. Further more, mycotoxins will residue in the products and tissues of animals fed with diet contaminated with them. Recent researches indicated that digestive function of intestinal tract could be changed by mycotoxins, and that ducks were more sensitive to mycotoxins than other animals. In this study, the effect of corn naturally contaminated with mycotoxins on the the growth performance, physiological function of digestive tract and its possible mechanisms were evaluated. This study consisted of 4 experiments as followed.
Experiment 1 Effect of corn naturally contaminated with mycotoxins on growth performance and intestinal morphology of ducks
This experiment was conducted to study the effect of corn naturally contaminated with mycotoxins on the growth performance and intestinal morphology for ducks.192 1-d-old Charry Vally ducks were randomly allotted by body weight to 2 groups with 6 replicates of 16 ducks each (8 males and 8 females).Group 1 (control) was fed the conventional diet with the corn free of mycotoxins, and group 2 was fed the diet with the corn naturally contaminated with mycotoxins with excessive content of AFB1. The trial lasted for 28 d. At d 0,14 and 28 of experiment, the ducks were weighed and consumption of feed by replicate was calculated. At d 28, jejunum was collected from 2 ducks in each replicate to determine the intestinal morphology. The results showed that diet with the corn naturally contaminated with mycotoxins resulted in the increase of mortality and decrease of growth performance of ducks. Compared with the control group, during d 0-28, the average daily gain (ADG) and average daily feed intake (ADFI) of mycotoxins contaminated corn group significantly reduced by 19.96%(p<0.01) and 18.50%(p< 0.05) respectively, and the d 0-14 F/G (feed to gain ratio)decreased significantly (p<0.01). Jejunum villus height the ducks fed mycotoxins contaminated corn diet increased by 22.11%(p<0.O5), crypt depth decreased by 32.69%(p<0.05), the villus height to crypt depth ratio (V/C) enhanced significantly(p<0.01) and the thickness of intestine wall tended to increase. The pathological changes were also found in these ducks fed diet with moldy corn, including hydrocardia, phaeo-pigment deposition in liver, and focal necrosis, desmoplasia, lympholeukocyte and adipocyte increasing in hepatic tissue. The results indicated that corn naturally contaminated with mycotoxins could decrease growth performance and increase the mortality, induce pathological changes in liver, and change intestinal morphology in ducks.
Experiment 2 Effect of different levels of corn contaminated with mycotoxins on growth performance and digestive physiology in ducks
In order to study the effect of corn naturally contaminated with mycotoxins (predominantly with AFB-1) on growth performance, intestinal development and digestive enzyme activities in ducks,384 ducks were allotted by body weight into 3 treatments with 8 replicates per treatment and 16 ducks(8 males and 8 females) per replicate. Treatment 1 (control) was fed diet with normal corn, and treatment 2 and 3 were fed the diets with contaminated corn substituted for normal com by 50% and 100% respectively. The trial lasted for 5 weeks. Growth performance, the digestive organ index and intestinal morphology at d 14 and 35, contents of DNA, RNA, protein of intestinal mucosa at d 14 and activities of digestive enzymes in brush border of jejunum and pancreas were determined. The results showed that ADFI, ADG and F/G linearly reduced (p< 0.001), and mortality linearly increased (p< 0.001) with increment of moldy com. The mortality in 100% contaminated corn group was 31.25%. At d 14 and 35, duodenum index, jejunum index and ileum index were increased (p< 0.0001) significantly, and so did the liver index at d 35. With the increasing levels of moldy corn, the V W (p<0.05), VH (p< 0.001) and VASA(p< 0.01) of jejunum mucosa increased at d 14, the CD (p< 0.01), VW (p=0.10), VH (p<0.05) and VASA (p<0.05) of duodenum mucosa also increased at d 35, content of DNA in duodenum and jejunum mucosa increased at d 14, which was opposite to that of RNA. In addition, the increment of addition levels of moldy com enhanced the activities of alkaline phosphatase (p<0.05), leucine aminopeptidase (p<0.05), trypsinase (p< 0.05) and chymotrypsin (p<0.001) at 14 d. The results indicated that the corn naturally contaminated predominantly with AFB1 could depress the feed intake, decrease body weight gain, increase mortality and reduce F/G through the changes in intestinal development and digestive enzymes activities.
Experiment 3 Effect of corn contaminated with mycotoxins on the rate of chyme to pass through the digestive tract and the digestibility of nutrients
This experiment was conducted to study the effect of corn naturally contaminated with mycotoxins on the rate of chyme to pass through the digestive tract and digestibility of nutrients. 384 ducks were randomly assigned to 3 dietary treatments with 8 replicates per treatment and 16 ducks (8 males and 8 females) per replicate. Group 1 was fed diet with normal corn (control), group 2 was fed diet with contaminated corn (contaminated), group 3 was pair-fed group to receive the same diet as control and the same amount of feed as the contaminated ducks (pair-fed). The trial lasted for 14 d. The growth performance, digestibility of nutrients and the rate of chyme to pass through the digestive tract were determined. Compared with control, on d 0-7,8-14 and 0-14, ADFI of contaminated corn group ducks was reduced by 23.55%(p< 0.01),47.56%(p< 0.01) and 43.23%(p<0.01) respectively, ADG was decreased by 31.47%(p<0.01),41.57%(p<0.01) and 38.84%(p< 0.01) respectively, however, compared with pair-fed group, ADG of contaminated group was improved by 4.63%、18.25% and 13.83%(p< 0.01) respectively. Compared with control and pair-fed group, d 0-14 F/G was reduced 4.90%(p< 0.01) and 5.56%(p< 0.01).digestibilities of DM were increased by 3.78%(p< 0.01) and 3.75%(p< 0.01), CP was increased by 2.65% and 4.63%(p< 0.01), total amino acid was increased by 3.9%(p< 0.01) and 3.98%(p< 0.01), Furthermore, the retention time of chyme in segment of digestive tract was obviously prolonged in contaminated group. The retention time in lower segment of esophagus and jejunum were prolonged by 37.78%(p< 0.05) and 55.94%(p< 0.05) respectively. The results indicated that corn naturally contaminated with mycotoxins could depress feed intake, decrease body weight gain and. increase digestibility of nutrients, dercrease F/G by reducing the rate of chyme to pass through the digestive tract.
Experiment 4. Effects of corn contaminated with mycotoxins on serum hormon levels and gene expression of Na+, K+-ATPase in ducks
256 ducks were randomly allotted by body weight into 2 treatment groups with 8 replicates per treatment and 16 ducks (8 males and 8 females) per replicate to explore the mechanism for the effect of corn contaminated with mycotoxins on the digestive physiology. The trial lasted for 4 weeks. At d 14,28,2 ducks (1 male and 1 female) of average weight in each replication were selected to collect 4 mL blood from jugular vein for determining contents of neuropeptide Y(NPY), ghrelin, leptin, CCK-8, insulin and IGF-I in serum,and then to be killed by cervical dislocation to collect 2-cm jejunum sample(first quarter) for determining relative expression of Na+, K+-ATPase gene. The content of leptin (p< 0.01)and IGF-I (p< 0.01) in serum at d 14 and 28 increased significantly when ducks fed diet with moldy corn, while the content of NPY at d 28 decreased significantly (p< 0.05) Furthermore, compared with control, the relative expression of Na+, K+-ATPase gene tended to increase. The results indicated that the decrease of NPY and increase of leptin and CCK-8 content in serum of ducks fed diet with corn naturally polluted with mycotoxins could lead to the depression of feed intake and growth performance. At the same time, the increase of IGF-I content could alleviate the lesion brought from mycotoxins to keep the health of ducks.
Implication
The results indicated that diet with corn naturally contaminated with mycotoxins (predominantly with AFB1) would increase the ducks mortality, decrease the feed intake and growth performance, and improve the feed efficiency. Futhermore, it was explored that the decrease of NPY and increase of Leptin content in serum were the main reason for depression of feed intake, and the decrease of feed intake was the main reason for depression of growth performance. At the same time, it was discovered that diet with corn naturally contaminated with mycotoxins (predominantly with AFB1) would increase the content of DNA in duodenum and jejunum mucosa and reduce that of RNA, improve VW, VH and VASA of jejunum, enhance the activities of alkaline phosphatase, leucine aminopeptidase, trypsinase and chymotrypsin, and prolong the retention time of chyme in digestive tract. It was concluded that diet naturally contaminated with mycotoxins had complicated effects on digestive end endocrine system of Ducks.
试验一自然霉变玉米对肉鸭生产性能和肠道形态结构的影响
本试验旨在研究以自然霉变玉米配制饲粮对肉鸭生产性能和肠道形态结构的影响。试验设2个处理,处理1为正常玉米对照组,处理2以霉变玉米(AFB1超标)100%替代正常玉米。每个处理6个重复,每个重复16只1日龄健康樱桃谷肉鸭(公母各半)。试验期4w。在试验的第0d、14d和28d按重复称肉鸭体重,并计算耗料；试验第28d每重复选2只肉鸭屠宰,采集空肠组织观测肠道形态结构。试验结果显示,自然霉变玉米组死亡率极显著升高(p<0.01),生产性能显著下降：与对照组相比,0-28d日增重和日采食量分别降低了19.96%(p<0.01)和18.50%(p<0.05),0-28d料重比略低(p>0.05)而0-14d料重比极显著降低(p<0.01)；自然霉变玉米组肉鸭肠绒毛高度较正常玉米组提高了22.11%(p<0.05),隐窝深度降低了32.69%(p<0.05),绒毛高度/隐窝深度极显著提高(p<0.01),肠壁厚略有提高(p>0.05)；自然霉变玉米组肉鸭发生心包积水、肝脏褐色素沉积及肝组织灶性坏死、结缔组织增生,淋巴细胞和脂肪细胞增多等病变。结果表明：摄食含AFB1超标自然霉变玉米的饲粮将导致肉鸭死亡率升高和生产性能降低,导致肝脏等内脏发生病变,并明显影响肠道形态结构。
试验二自然霉变玉米对肉鸭生产性能、肠道发育及消化酶活性的影响
本试验旨在研究肉鸭摄食以污染多种霉菌毒素且AFB1超标的自然霉变玉米配制的饲粮后,生产性能、肠道发育和消化酶活性的变化。试验采用单因子设计,共3个处理组,即用自然霉变玉米分别按0%(对照组)、50%和100%比例替代饲粮中正常玉米,每个处理8个重复,每个重复16只肉鸭,公母各半。试验期5w。试验考察了肉鸭各阶段的生产性能,观测了第14 d和35 d的消化器官指数和肠道形态,测定了第14 d的肠黏膜RNA和蛋白质含量,以及第14 d的空肠刷状缘酶与胰腺酶活性。结果显示：在整个试验期内,随着霉变玉米在饲粮中用量增加,肉鸭的ADFI、ADG、F/G均呈线性降低(p<0.001),死亡率线性升高(p<0.001),100%霉变玉米组鸭的死亡率达31.25%;第14d和35d的十二指肠指数、空肠指数、回肠指数及第35d的肝脏指数均较其余两组极显著增加(p<0.001)；随霉变玉米的用量增加,第14d空肠粘膜的绒毛宽度(VW)(p<0.05)、绒毛高度(VH)(p<0.001)和绒毛表面积(VASA)(p<0.01)增加,第35d十二指肠粘膜的隐窝深度(CD)(p<0.01).VW(p=0.10).VH(p<0.05)和VASA(p<0.05)增加,第14d十二指肠和空肠粘膜的DNA含量增加而RNA含量降低,第14d空肠刷状缘碱性磷酸酶活性(p<0.05)和亮氨酸氨肽酶活性(p<0.05)以及胰腺胰蛋白酶活性(p<0.05)和糜蛋白酶活性(p<0.001)升高。结果表明：自然霉变玉米饲粮(AFB1超标)使肉鸭采食受到抑制,增重减慢,死亡率增加,改变肉鸭肠道发育和消化酶活性。
试验三自然霉变玉米对肉鸭食糜通过速率和养分消化率的影响
本试验旨在研究以自然霉变玉米配制的饲粮对肉鸭食糜通过速率和养分消化率的影响。试验设3个处理,处理1为正常玉米组,处理2为霉变玉米组,处理3为采食配对组,即肉鸭采食的饲粮同处理1而采食的量同处理2,每个处理8个重复,每个重复16只肉鸭。试验期14d。试验考察了肉鸭的生产性能、养分消化率和食糜流通速率。结果显示：在0-7d、8-14d和0-14d,与正常玉米组相比,自然霉变玉米组肉鸭ADFI分别降低了23.55%(p<0.01)、47.56%(p<0.01)和43.23%(p<0.01),ADG分别降低了31.47%(p<0.01)、41.57%(p<0.01)和38.84%(p<0.01),与采食配对组相比,自然霉变玉米组ADG分别增加4.63%、18.25%和13.83%(p<0.01)；与正常玉米组和采食配对组相比,霉变玉米组全期F/G分别降低4.90%(p<0.01)和5.56%(p<0.01),DM消化率分别提高3.78%(p<0.01)和3.75%(p<0.01),CP消化率分别提高2.65%和4.63%(p<0.01),总氨基酸消化率分别提高3.9%(p<0.01)和3.98%(p<0.01)。同时,自然霉变玉米组食糜在消化道各段的停留时间延长了,与正常玉米组相比,食管膨大部和空肠停留时间分别延长37.78%(p<0.05)和55.94%(p<0.05)。结果表明：以自然霉变玉米配制饲粮(AFB1超标)饲喂肉鸭,会抑制肉鸭采食,降低肉鸭增重,同时延长食糜在消化道的停留时间,提高养分消化率、降低料重比。
试验四自然霉变玉米对肉鸭血清激素水平和肠组织Na+,K+-ATPase基因表达的影响
为了进一步探明自然霉变玉米影响肉鸭摄食及消化生理的机制,本试验研究了以自然霉变玉米配制的饲粮对肉鸭血清采食调控相关激素水平和肠组织Na+,K+-ATPase基因表达的影响。试验设2个处理,处理1为正常玉米饲粮组,处理2饲粮则以自然霉变玉米(AFB1超标)100%替代正常玉米配制,每个处理8个重复,每个重复16只1日龄健康肉鸭。试验期28天。第14 d和第28 d,分别从每个重复中选取2只肉鸭(公母各一只),颈静脉采血4ml,测定血清中神经肽Y (NPY)、Ghrelin、瘦素(Leptin)、八肽胆囊收缩素(CCK-8)、胰岛素、胰岛素样生长因子-1(IGF-1)的含量。第14 d采血后处死肉鸭,采集空肠前段(前1/4处)组织,检测Na+,K+-ATPase基因相对表达量。结果显示：自然霉变玉米组肉鸭14d和28d的血清中IGF-1和Leptin的含量皆极显著提高(p<0.01),而第28d血清NPY含量降低(p<0.05)；肉鸭空肠组织Na+,K+-ATPase基因相对表达量较对照组增加了23%(p>0.05)。结果表明：肉鸭采食自然霉变玉米饲粮(AFB1超标)后,可能通过提高血清Leptin并降低血清NPY含量,进而抑制肉鸭采食；可能通过提高血清IGF-1含量和肠组织Na+, K+-ATPase基因表达量来影响养分吸收和饲料效率,缓解霉菌毒素的危害。
综上所述,以自然霉变玉米配制的饲粮饲喂肉鸭会导致死淘亡率升高,采食量、增重和料重比降低,饲料养分表观消化率提高。进一步分析发现,自然霉变玉米导致的增重下降主要是通过降低肉鸭的采食量所致,其机制与血清Leptin含量提高而NPY含量降低有关；而料重比较低的原因是饲料养分消化利用率提高,其机制与十二指肠和空肠粘膜的DNA含量增加、RNA含量降低,绒毛高度、宽度和表面积增加,空肠组织Na+, K+-ATPase基因表达量上升,空肠刷状缘的碱性磷酸酶和亮氨酸氨肽酶活性提高,胰腺胰蛋白酶和糜蛋白酶活性增强,以及食糜在消化道的通过速率减慢等因素的综合作用有关。结果提示,摄食含AFB1超标自然霉变玉米的饲粮对肉鸭消化系统和内分泌系统产生较复杂的影响,进而影响生产性能。
Feed contaminated with mycotoxins is harmful to animals. It reduces growth and depresses the immunity of animals. These effects have serious economic implications for animal producers. Further more, mycotoxins will residue in the products and tissues of animals fed with diet contaminated with them. Recent researches indicated that digestive function of intestinal tract could be changed by mycotoxins, and that ducks were more sensitive to mycotoxins than other animals. In this study, the effect of corn naturally contaminated with mycotoxins on the the growth performance, physiological function of digestive tract and its possible mechanisms were evaluated. This study consisted of 4 experiments as followed.
Experiment 1 Effect of corn naturally contaminated with mycotoxins on growth performance and intestinal morphology of ducks
This experiment was conducted to study the effect of corn naturally contaminated with mycotoxins on the growth performance and intestinal morphology for ducks.192 1-d-old Charry Vally ducks were randomly allotted by body weight to 2 groups with 6 replicates of 16 ducks each (8 males and 8 females).Group 1 (control) was fed the conventional diet with the corn free of mycotoxins, and group 2 was fed the diet with the corn naturally contaminated with mycotoxins with excessive content of AFB1. The trial lasted for 28 d. At d 0,14 and 28 of experiment, the ducks were weighed and consumption of feed by replicate was calculated. At d 28, jejunum was collected from 2 ducks in each replicate to determine the intestinal morphology. The results showed that diet with the corn naturally contaminated with mycotoxins resulted in the increase of mortality and decrease of growth performance of ducks. Compared with the control group, during d 0-28, the average daily gain (ADG) and average daily feed intake (ADFI) of mycotoxins contaminated corn group significantly reduced by 19.96%(p<0.01) and 18.50%(p< 0.05) respectively, and the d 0-14 F/G (feed to gain ratio)decreased significantly (p<0.01). Jejunum villus height the ducks fed mycotoxins contaminated corn diet increased by 22.11%(p<0.O5), crypt depth decreased by 32.69%(p<0.05), the villus height to crypt depth ratio (V/C) enhanced significantly(p<0.01) and the thickness of intestine wall tended to increase. The pathological changes were also found in these ducks fed diet with moldy corn, including hydrocardia, phaeo-pigment deposition in liver, and focal necrosis, desmoplasia, lympholeukocyte and adipocyte increasing in hepatic tissue. The results indicated that corn naturally contaminated with mycotoxins could decrease growth performance and increase the mortality, induce pathological changes in liver, and change intestinal morphology in ducks.
Experiment 2 Effect of different levels of corn contaminated with mycotoxins on growth performance and digestive physiology in ducks
In order to study the effect of corn naturally contaminated with mycotoxins (predominantly with AFB-1) on growth performance, intestinal development and digestive enzyme activities in ducks,384 ducks were allotted by body weight into 3 treatments with 8 replicates per treatment and 16 ducks(8 males and 8 females) per replicate. Treatment 1 (control) was fed diet with normal corn, and treatment 2 and 3 were fed the diets with contaminated corn substituted for normal com by 50% and 100% respectively. The trial lasted for 5 weeks. Growth performance, the digestive organ index and intestinal morphology at d 14 and 35, contents of DNA, RNA, protein of intestinal mucosa at d 14 and activities of digestive enzymes in brush border of jejunum and pancreas were determined. The results showed that ADFI, ADG and F/G linearly reduced (p< 0.001), and mortality linearly increased (p< 0.001) with increment of moldy com. The mortality in 100% contaminated corn group was 31.25%. At d 14 and 35, duodenum index, jejunum index and ileum index were increased (p< 0.0001) significantly, and so did the liver index at d 35. With the increasing levels of moldy corn, the V W (p<0.05), VH (p< 0.001) and VASA(p< 0.01) of jejunum mucosa increased at d 14, the CD (p< 0.01), VW (p=0.10), VH (p<0.05) and VASA (p<0.05) of duodenum mucosa also increased at d 35, content of DNA in duodenum and jejunum mucosa increased at d 14, which was opposite to that of RNA. In addition, the increment of addition levels of moldy com enhanced the activities of alkaline phosphatase (p<0.05), leucine aminopeptidase (p<0.05), trypsinase (p< 0.05) and chymotrypsin (p<0.001) at 14 d. The results indicated that the corn naturally contaminated predominantly with AFB1 could depress the feed intake, decrease body weight gain, increase mortality and reduce F/G through the changes in intestinal development and digestive enzymes activities.
Experiment 3 Effect of corn contaminated with mycotoxins on the rate of chyme to pass through the digestive tract and the digestibility of nutrients
This experiment was conducted to study the effect of corn naturally contaminated with mycotoxins on the rate of chyme to pass through the digestive tract and digestibility of nutrients. 384 ducks were randomly assigned to 3 dietary treatments with 8 replicates per treatment and 16 ducks (8 males and 8 females) per replicate. Group 1 was fed diet with normal corn (control), group 2 was fed diet with contaminated corn (contaminated), group 3 was pair-fed group to receive the same diet as control and the same amount of feed as the contaminated ducks (pair-fed). The trial lasted for 14 d. The growth performance, digestibility of nutrients and the rate of chyme to pass through the digestive tract were determined. Compared with control, on d 0-7,8-14 and 0-14, ADFI of contaminated corn group ducks was reduced by 23.55%(p< 0.01),47.56%(p< 0.01) and 43.23%(p<0.01) respectively, ADG was decreased by 31.47%(p<0.01),41.57%(p<0.01) and 38.84%(p< 0.01) respectively, however, compared with pair-fed group, ADG of contaminated group was improved by 4.63%、18.25% and 13.83%(p< 0.01) respectively. Compared with control and pair-fed group, d 0-14 F/G was reduced 4.90%(p< 0.01) and 5.56%(p< 0.01).digestibilities of DM were increased by 3.78%(p< 0.01) and 3.75%(p< 0.01), CP was increased by 2.65% and 4.63%(p< 0.01), total amino acid was increased by 3.9%(p< 0.01) and 3.98%(p< 0.01), Furthermore, the retention time of chyme in segment of digestive tract was obviously prolonged in contaminated group. The retention time in lower segment of esophagus and jejunum were prolonged by 37.78%(p< 0.05) and 55.94%(p< 0.05) respectively. The results indicated that corn naturally contaminated with mycotoxins could depress feed intake, decrease body weight gain and. increase digestibility of nutrients, dercrease F/G by reducing the rate of chyme to pass through the digestive tract.
Experiment 4. Effects of corn contaminated with mycotoxins on serum hormon levels and gene expression of Na+, K+-ATPase in ducks
256 ducks were randomly allotted by body weight into 2 treatment groups with 8 replicates per treatment and 16 ducks (8 males and 8 females) per replicate to explore the mechanism for the effect of corn contaminated with mycotoxins on the digestive physiology. The trial lasted for 4 weeks. At d 14,28,2 ducks (1 male and 1 female) of average weight in each replication were selected to collect 4 mL blood from jugular vein for determining contents of neuropeptide Y(NPY), ghrelin, leptin, CCK-8, insulin and IGF-I in serum,and then to be killed by cervical dislocation to collect 2-cm jejunum sample(first quarter) for determining relative expression of Na+, K+-ATPase gene. The content of leptin (p< 0.01)and IGF-I (p< 0.01) in serum at d 14 and 28 increased significantly when ducks fed diet with moldy corn, while the content of NPY at d 28 decreased significantly (p< 0.05) Furthermore, compared with control, the relative expression of Na+, K+-ATPase gene tended to increase. The results indicated that the decrease of NPY and increase of leptin and CCK-8 content in serum of ducks fed diet with corn naturally polluted with mycotoxins could lead to the depression of feed intake and growth performance. At the same time, the increase of IGF-I content could alleviate the lesion brought from mycotoxins to keep the health of ducks.
Implication
The results indicated that diet with corn naturally contaminated with mycotoxins (predominantly with AFB1) would increase the ducks mortality, decrease the feed intake and growth performance, and improve the feed efficiency. Futhermore, it was explored that the decrease of NPY and increase of Leptin content in serum were the main reason for depression of feed intake, and the decrease of feed intake was the main reason for depression of growth performance. At the same time, it was discovered that diet with corn naturally contaminated with mycotoxins (predominantly with AFB1) would increase the content of DNA in duodenum and jejunum mucosa and reduce that of RNA, improve VW, VH and VASA of jejunum, enhance the activities of alkaline phosphatase, leucine aminopeptidase, trypsinase and chymotrypsin, and prolong the retention time of chyme in digestive tract. It was concluded that diet naturally contaminated with mycotoxins had complicated effects on digestive end endocrine system of Ducks.
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