术苦芩总多糖对湿热泄泻仔猪肠道菌群和免疫功能的影响

旨在探究术苦芩总多糖(ZKQPs)对湿热泄泻仔猪肠道菌群和免疫功能的影响。通过HPLC分析ZKQPs的单糖组成,体外抑菌试验研究ZKQPs体外对常见肠道致病菌的直接影响,同时建立仔猪腹泻模型,用白头翁散(PP)及3种剂量的ZKQPs (25、50和75 mg·kg^-1)治疗腹泻仔猪,治疗结束后,收集小肠各段组织、盲肠内容物,对菌落进行鉴定与计数;通过16S rDNA高通量测序分析肠道菌群的变化,测定小肠免疫相关细胞因子mRNA表达。结果显示:ZKQPs一级结构主要由鼠李糖、葡萄糖、半乳糖和木糖构成。在体外试验中,大肠杆菌、沙门菌、猪源产气荚膜梭菌肠道常见有害菌对ZKQPs敏感,并在一定范围内呈现浓度依赖性。动物试验中,ZKQPs有效促进了腹泻仔猪肠道乳酸杆菌的生长,对致病大肠杆菌生长有较强的抑制作用,其中,50 mg·kg^-1ZKQPs影响最为显著。16S rDNA高通量测序结果表明,ZKQPs调节了腹泻仔猪菌群Alpha多样性,对肠道菌群在纲和属水平上有显著影响,腹泻仔猪芽胞杆菌纲、乳酸杆菌属相对丰度显著下降,梭菌纲相对丰度显著上升;ZKQPs治疗后芽胞杆菌纲、乳酸杆菌属相对丰度均显著上升且高于正常值,梭菌纲相对丰度极显著下降,显著改善了腹泻仔猪肠道菌群结构。仔猪腹泻时,小肠各段Th1细胞因子IL-2 mRNA水平上升(P<0.01),Th2细胞因子IL-4、IL-5 mRNA水平均下降;ZKQPs治疗后,腹泻仔猪小肠各段IL-2 mRNA水平下降,IL-4、IL-5 mRNA水平普遍上升,且不同细胞因子mRNA水平在小肠不同位置变化有所差异,50和75 mg·kg^-1ZKQPs治疗后对其影响最为显著,综合分析50 mg·kg^-1ZKQPs治疗腹泻仔猪对细胞因子mRNA水平影响最大。综上表明,ZKQPs在体外对常见肠道致病菌生长具有良好的抑制作用,可以改变腹泻仔猪肠道微生物群的丰富度和多样性,提高乳酸杆菌属的相对丰度,并改善菌群结构,同时调节肠道免疫反应,有效增强了仔猪抗腹泻能力。     

术苦芩总多糖对湿热泄泻仔猪小肠杯状细胞数量以及MUC-2和ITF-3 mRNA转录的影响

为探讨术苦芩总多糖(ZKQPs)对湿热泄泻仔猪小肠修复作用的影响,将60头断奶仔猪分为空白对照组(n=10)造模组(n=50),造模组采用高温高湿环境配合高脂配合饲料等建立湿热泄泻模型,进一步将造模成功后的50头仔猪随机分为模型组、阳性药物组(白头翁散)、ZKQPs高、中、低剂量组(n=10),分别拌料给药,连用7d。采用过碘酸雪夫染色(PAS)结合显微镜观察小肠组织杯状细胞(GC),利用RT-PCR和ELISA分别检测湿热泄泻仔猪小肠黏液中黏蛋白-2(MUC-2)和肠三叶因子(ITF-3)的转录和含量变化。试验结果:1)模型组仔猪十二指肠、空肠、回肠中GC数量均减少,与空白组比较,差异显著(P<0.05);与模型组比较,ZKQPs高剂量组、中剂量组和阳性药物组仔猪十二指肠、空肠、回肠中GC数量均增加,ZKQPs高剂量组、阳性药物组仔猪十二指肠、空肠、回肠和ZKQPs中剂量组仔猪十二指肠、回肠GC的数量差异显著(P<0.05)。2)与空白组比较,模型组仔猪十二指肠、空肠、回肠中MUC-2mRNA的转录量和含量均降低,差异显著(P<0.05或P<0.01)。与模型组比较,ZKQPs各剂量组和阳性药物组仔猪十二指肠、空肠、回肠中MUC-2mRNA的转录量和含量均升高,ZKQPs高剂量组和阳性药物组仔猪十二指肠、空肠、回肠MUC-2mRNA的转录量和含量差异显著(P<0.05或P<0.01)。3)与空白组比较,模型组仔猪十二指肠、空肠、回肠中ITF-3mRNA的转录量和含量均降低,差异极显著(P<0.01)。与模型组比较,ZKQPs高剂量组、中剂量组和阳性药物组仔猪十二指肠、空肠、回肠中ITF-3mRNA的转录量和含量均升高,ZKQPs高剂量组、中剂量组和阳性药物组仔猪空肠、回肠中ITF-3mRNA的转录量和含量差异显著(P<0.05或P<0.01)。ZKQPs可提高湿热泄泻仔猪小肠GC的数量以及MUC-2和ITF-3mRNA的转录,促进湿热泄泻仔猪小肠内GC分泌物MUC-2和ITF-3分泌。     

术苦芩颗粒对靶动物仔猪的安全性试验

为了评价术苦芩颗粒对靶动物猪的安全性,选取60头健康的20日龄荣昌猪,随机分为4组,每组15头。各试验组分别以1倍(5 g/kg)、3倍(15 g/kg)、5倍(25 g/kg)推荐剂量的术苦芩颗粒连续经口灌胃给药14 d,观察临床症状、增重、血液学和组织病理学变化。结果显示,各组间的血液生理、生化指标无明显差异,组织无明显病理学病变,中剂量组仔猪日增重显著增加(P0.05)。试验表明,术苦芩颗粒安全范围大,在仔猪体内耐受性高,以5倍推荐剂量灌胃14 d对靶动物猪安全。     

加味葛根芩连汤对湿热泄泻仔猪肠道炎症和损伤修复的作用

通过用加味葛根芩连汤治疗仔猪的湿热泄泻,探讨其对湿热泄泻仔猪肠道的修复作用.选取8～15日龄未断奶仔猪(大白×长白)30头,其中健康仔猪6头,为对照组;有粪色黄褐而臭、小便短赤、舌质红等症状的泄泻仔猪24头,分为湿热泄泻组、加味葛根芩连汤低剂量组、加味葛根芩连汤中剂量组和加味葛根芩连汤高剂量组,每组6头.对照组和湿热泄...     

术苦芩颗粒对湿热泻痢仔猪血清D-木糖、淀粉酶及脂肪酶的影响

为了观察中药复方术苦芩颗粒对湿热泻痢仔猪血清D-木糖、淀粉酶及脂肪酶的影响,将60头仔猪随机分为6组:空白组、模型组、阳性药物组、术苦芩颗粒高、中、低剂量组,每组10头;利用湿热环境、细菌侵袭等恶劣环境人工复制仔猪湿热泻痢模型;观察猪只体征变化,采用间苯三酚法测血清D-木糖、ELISA法测脂肪酶与淀粉酶。结果湿热泻痢仔猪血清D-木糖含量、淀粉酶活性及脂肪酶活性改变,术苦芩颗粒可明显改善其临床症状,治愈湿热泻痢仔猪,促进仔猪D-木糖吸收和恢复淀粉酶、脂肪酶的活性。表明术苦芩颗粒可有效治疗仔猪湿热泻痢,并改善仔猪小肠的消化吸收功能。     

加味葛根芩连汤对湿热泄泻仔猪肠道炎症和损伤修复的作用

通过用加味葛根芩连汤治疗仔猪的湿热泄泻,探讨其对湿热泄泻仔猪肠道的修复作用。选取8~15日龄未断奶仔猪（大白×长白）30头,其中健康仔猪6头,为对照组;有粪色黄褐而臭、小便短赤、舌质红等症状的泄泻仔猪24头,分为湿热泄泻组、加味葛根芩连汤低剂量组、加味葛根芩连汤中剂量组和加味葛根芩连汤高剂量组,每组6头。对照组和湿热泄泻组仔猪灌服生理盐水,药物组仔猪灌服加味葛根芩连汤（按生药量,低、中、高剂量组药物添加量分别为2.5、5.0和10.0 g·d^-1）,连续5 d。在服药的第0、3、5天记录各组仔猪的粪便评分。取对照组、湿热泄泻组、加味葛根芩连汤中剂量组仔猪十二指肠、空肠、回肠和结肠进行组织切片和H.E.染色,取十二指肠进行增殖性细胞核抗原（PCNA）免疫组化染色;用荧光定量PCR检测对照组、湿热泄泻组、加味葛根芩连汤中剂量组仔猪空肠TLR4、TNF-α、IL-1β和IL-6 mRNA表达量变化。湿热泄泻严重破坏了仔猪的十二指肠、空肠、回肠和结肠结构,尤其是十二指肠和空肠,十二指肠隐窝处PCNA表达量显著升高（P<0.05）。与对照组相比,湿热泄泻组仔猪空肠TLR4（P<0.05）、TNF-α（P<0.01）、IL-1β（P<0.01）和IL-6（P<0.05）mRNA表达量显著升高。相比于湿热泄泻组,加味葛根芩连汤显著降低了仔猪的粪便评分（P<0.05）。中剂量加味葛根芩连汤组仔猪的小肠和结肠的组织结构得到显著改善,十二指肠的肠腔中有不断脱落的绒毛团,绒毛中的PCNA表达量极显著升高（P<0.01）。与湿热泄泻组相比,加味葛根芩连汤中剂量组空肠TLR4（P<0.05）、TNF-α（P<0.01）、IL-1β（P<0.01）和IL-6（P<0.05）mRNA表达量显著降低。综上表明,加味葛根芩连汤可通过促进隐窝干细胞的增殖和降低炎症反应对湿热泄泻造成的仔猪肠道损伤进行修复。     

苦芩总多糖对免疫低下小鼠免疫功能的影响

为了探索苦芩总多糖对免疫低下小鼠免疫功能的影响,试验通过对苦芩中总多糖的提取,并将所提取总多糖对免疫低下小鼠进行一系列免疫相关试验,如碳粒廓清指数测定试验、免疫器官指数测定试验、淋巴细胞转化试验、血清溶血素的测定试验。结果表明,苦芩及其总多糖均能显著提高免疫抑制小鼠的吞噬指数α,脾脏指数和胸腺指数,同时还能显著提高免疫抑制小鼠血清溶血素水平以及T、B淋巴细胞转化率。表明苦芩总多糖可以通过调节机体的免疫系统从而增强机体的免疫功能。     

基于网络药理学探究苦参止痢颗粒治疗湿热泄泻的有效成分及作用机制

利用中药系统药理学数据库与分析平台(TCMSP)对苦参止痢颗粒3味中药的化学成分及其作用靶点进行挖掘,通过Uniport数据库标准化.从GeneCards数据库和OMIM数据库获取泄泻的相关基因,将苦参止痢颗粒与泄泻的基因靶点进行映射取交集,利用STRING数据库构建蛋白相互作用(PPI)网络,通过DAVID数据库进行...     

女贞子多糖对仔猪抗氧化及MAPK通路的影响

为了研究女贞子多糖对人工感染大肠杆菌K88仔猪氧化应激的保护作用,将实验猪分为四组:对照组、模型组、女贞子多糖组和女贞子多糖饲喂后攻毒组。试验运用血清生化、ELISA和RT-PCR等技术,通过检测血清和肝肾组织的抗氧化酶和氧化产物,仔猪肝脏氧化应激相关基因转录水平和MAPK信号关键蛋白的转录,评价女贞子多糖的抗氧化机制。结果显示,女贞子多糖可以显著降低由大肠杆菌引起的仔猪血清MDA含量的升高,调节肝肾组织中MDA和SOD的水平;女贞子多糖可以提高仔猪肝脏中CAT和CuZnSOD的mRNA转录水平;正常情况下,女贞子多糖对仔猪肝脏TNF-α的mRNA转录表达无影响,但可以降低大肠杆菌攻毒后的转录水平;女贞子多糖抑制MAPK通路主要依靠调控ERK1起作用。结果表明,女贞子多糖可以通过调节肝脏抗氧化指标保护大肠杆菌引起的仔猪氧化应激反应,其作用分子机制可能与下调MAPK相关信号转录水平有关。     

中药"泄泻康"对仔猪泄泻的预防试验初报

仔猪泄泻是仔猪出生至断乳前后临床常见的疾病之一。笔者将盱眙县师皇动物科技研究所研制的纯中药“泄泻康”添加到乳猪料中饲喂仔猪。证明“泄泻康”对仔猪泄泻,尤其是断乳前后泄泻有比较明显的预防作用。     

腹泻仔猪肠道沙门氏菌耐药性研究

为了解贵州省规模化养殖场腹泻仔猪肠道沙门菌耐药表型与耐药基因型相关性,本试验采集了贵州省7个规模化养猪场的腹泻仔猪肠道样本进行沙门菌分离鉴定,并对分离的沙门菌用药敏纸片法进行耐药表型分析,利用PCR方法对分离菌进行耐药基因扩增。结果显示:共分离鉴定沙门菌21株;药敏试验表明所鉴定的沙门菌对抗菌药物的耐药率从高到低依次是卡那霉素、磺胺异口恶唑和诺氟沙星等,且均为多重耐药株,耐药种类达8种以上;耐药基因与耐药表型分析结果显示分离的沙门菌β-内酰胺类耐药基因与耐药表型符合率高达100%,喹诺酮耐药基因与耐药表型符合率为62.5%,氨基糖苷耐药基因符合率为73.3%,磺胺类耐药基因符合率81.8%。研究结果对合理使用药物控制仔猪沙门菌感染提供了理论依据。     

植物多糖对断奶仔猪小肠黏膜形态及肠黏膜屏障功能的影响

旨在探讨断奶仔猪饲粮中单独或联合添加黄芪多糖、牛膝多糖及白术多糖对其小肠黏膜形态及肠黏膜屏障功能的影响。选取256头35日龄的杜长大三元杂交断奶仔猪,随机分为8个组,每组4个重复,每个重复8头猪,采用三因素两水平(2×2×2)试验设计,饲粮中黄芪多糖、白术多糖和牛膝多糖的添加水平均为0,800mg/kg,试验共进行28d。结果显示:植物多糖能显著提高断奶仔猪十二指肠、空肠和回肠的黏膜绒毛高度,提高绒腺比(P0.05),3种多糖联用效果最适。与对照组相比,多糖组断奶仔猪回肠V/C比值显著提高(P0.05),各单一多糖组之间差异不显著(P0.05),3种多糖联合添加与两两添加及单一添加相比显著提高了V/C比值(P0.05)。多糖组断奶仔猪血清中二胺氧化酶(DAO)的活性、血清中内毒素(ET)含量及D-乳糖的含量均显著降低(P0.05),3种多糖联用效果最适。多糖能显著提高断奶仔猪肠道黏膜Occludin、Claudin和ZO-1的基因表达量(P0.05)。单一添加黄芪多糖、白术多糖及牛膝多糖能显著提高断奶仔猪肠道钠-葡萄糖共转运载体1(SGLT1)的mRNA相对表达量(P0.05),对葡萄糖转运载体2(GLUT2)mRNA相对表达量有提高的趋势,但差异不显著(P0.05)。多糖两两联用及3种多糖联合添加能显著提高断奶仔猪肠道SGLT1和GLUT2mRNA相对表达量(P0.05)。结果表明,在断奶仔猪饲粮中添加植物多糖可显著改善断奶仔猪肠道黏膜形态结构和肠道的通透性,其机制可能与植物多糖改善断奶仔猪肠黏膜屏障功能作用有关。     

Dietary high protein-induced diarrhea and intestinal inflammation by activation of NF-κB signaling in piglets

The present study aimed to investigate whether inflammation-associated responses in piglets are induced by high protein (HP) through activating nuclear factor kappa B (NF-κB) signaling. Sixteen piglets (35 d of age, Duroc × [Landrace × Yorkshire], weaned at d 21, initial BW = 9.70 ± 0.11 kg) were allocated to 18% and 26% CP (HP group) at random, comprising 8 replicate pens per treatment. The piglets were slaughtered to collect intestinal tissues when apparent, persistent, and stable diarrhea syndromes happened (on d 12). No significant differences were observed in their growth performance (P > 0.05), but reduction by 19.11%, 25.31%, 23.64% of ADFI, ADG, and G:F, respectively was detected in the HP group. The HP group had greater (P = 0.002) diarrhea rates. Furthermore, dietary HP had lower ileal villus height (VH; P = 0.048), ratio of villus height to crypt depth (VH/CD ratio; P = 0.016), and colonic CD (P = 0.034), as well as had the trend (P = 0.075) to reduce the ileal villus absorptive area. Moreover, HP diets significantly elevated the goblet cell numbers in the ileal villi (P = 0.016) and colonic crypts (P < 0.001) and up-regulated (P = 0.012) the mRNA expression of mucin2 (Muc2) in the ileum. In addition, HP diets increased the myeloperoxidase concentration in the ileum (P = 0.002) and colon (P = 0.007) of piglets. Dietary HP significantly down-regulated the mRNA expression of tumor necrosis factor-α (TNF-α; P < 0.001) in the ileum, induced nitric oxide synthase (iNOS; P = 0.040) and interleukin-22 (IL-22; P = 0.008) in the colon, and inclined to down-regulate interleukin-1β (IL-1β; P = 0.076) expression in the colon. The relative protein abundance of Galectin-3 (P = 0.046) in the colon and the ratio of phosphorylation NF-κB to NF-κB (p–NF–κB/NF-κB ratio) in the ileum of HP piglets were also greater (P = 0.038). These results suggest that dietary HP may cause diarrhea in piglets by activating NF-κB signaling induced intestinal inflammation.     

无抗条件下从肠道健康方面控制断奶仔猪腹泻的策略

在过去的几十年里,抗菌化合物已被用于预防亚临床和临床疾病,促进断奶仔猪生长等方面。但人们越来越关注细菌耐药性、耐药菌株及其相关基因对人类健康的潜在影响。欧盟从2006年1月1日起禁止在猪和家禽生产中使用抗生素作为生长促进剂。此外,高锌和高铜等矿物质也具有促生长作用,但饲料中添加高锌、高铜会严重污染环境。因此,在不使用抗生素的条件下,有必要制定相应的营养和饲喂策略,以便控制与断奶过渡有关的问题。本文综述改善胃肠道结构、功能,促进断奶后仔猪生长的营养策略,重点介绍益生菌、益生元、有机酸、微量元素来源及水平。     

Tannic acid extracted from gallnut prevents post-weaning diarrhea and improves intestinal health of weaned piglets

This experiment was conducted to evaluate the effects of different levels of tannic acid (TA) on growth performance, diarrhea rate, nutrient digestibility and intestinal health in weaned piglets. A total of 180 weaned piglets (Duroc × Landrace × Yorkshire, 24 d of age, initial average BW = 7.77 ± 0.17 kg) were allotted to 5 groups (6 pigs/pen and 6 replicates/group) in a randomized complete block design according to their gender and body weight. Piglets were fed a basal diet, or the basal diet supplemented with 0.05%, 0.1%, 0.2% or 0.4% TA for 28 d. The supplementary levels of TA in the diets were obtained by adding tannalbin containing 51% TA and 40.17% protein. The results showed that, compared with the CON group, dietary TA did not affect ADFI, ADG or F:G, and linearly reduced (P < 0.01) the diarrhea rate and diarrhea index of piglets. There were no significant effects on apparent total tract digestibility (ATTD) in the 0.05%, 0.1% and 0.2% TA groups, while negative effects (P < 0.05) on apparent digestibility of crude protein and gross energy were observed in the 0.4% TA group. In addition, the nutrient digestibility of dry matter, crude protein and gross energy linearly decreased (P < 0.01) with the increase of TA dosage. Supplementation of TA increased (P < 0.05) the villus height of the duodenum and jejunum, as well as increased (P < 0.05) catalase (CAT) activity in serum. Dietary TA improved (P < 0.05) the Bacillus counts in cecal digesta. Further, TA significantly improved (P < 0.05) Bacillus counts and reduced (P < 0.05) the Escherichia coli counts in colonic digesta. The concentration of acetic acid, propionic acid, butyric acid and isovaleric acid in cecal digesta were significantly increased (P < 0.05). The mRNA expression level of zonula occludens-1 (ZO-1), zonula occludens-2 (ZO-2), and claudin-2 (CLDN-2) in the jejunum were greater (P < 0.05) in TA supplemented groups. The study showed that, compared to the control, TA prevented post-weaning diarrhea and improved intestinal health of weaned piglets, and the appropriate level of TA supplementation would be from 0.1% to 0.2%.     

黄芪多糖对断奶仔猪免疫功能及腹泻的影响

研究了在断奶应激模式下添加黄芪多糖 (APS)对断奶仔猪免疫功能及腹泻的影响。试验分 3组 :对照Ⅰ组 (2 8日龄不断奶 ) ,对照Ⅱ组(2 8日龄断奶 )以及试验组 (2 8日龄断奶 +APS)。结果 :试验组与对照Ⅱ组相比 ,在断奶后第 7、 1 4天淋巴细胞百分数分别增加 7 7% (P >0 0 5)、 1 8 2 % (P >0 0 5) ;中性粒细胞百分数分别减少了 1 5 2 % (P >0 0 5)、 2 8 6 % (P >0 0 5) ;断奶仔猪的SRBC抗体效价分别提高 0 47、1 49(P <0 0 5)个滴度 ;单核细胞增殖率分别提高 1 1 % (P >0 0 5)、 2 5 % (P <0 0 5)。腹泻率在 3组间差异不显著 ;APS添加组腹泻频率显著低于对照Ⅱ组 (P <0 0 5) ,与对照Ⅰ组相比 ,差异不显著     

Microscopic structure of the large intestinal mucosa in piglets during an antibiotic-associated diarrhea

Antibiotic-associated diarrhea (AAD) is caused by the treatments of broad-spectrum antimicrobials that seriously affect the activity and composition of the large intestinal microflora. The pathogenic bacteria or low concentration of short-chain fatty acids (SCFAs) has been repeatedly discussed in relation to AAD. Recently, we reported the detection of a large amount of succinate and lactate in the diarrheal feces in AAD-induced piglets. In this study, we investigated histologically the large intestinal mucosa in AAD-induced piglets, in which succinate and lactate were accumulated. AAD was induced in the piglets by an oral dose of polymyxin B sulfate (PL) or by an intra-muscular injection of enrofloxacin (ERFX). When the piglets were defecating diarrheal feces with a high concentration of succinate and/or lactate, the large intestine was removed and separated into four segments (cecum, gyri centripetales, gyri centrifugales, and rectum). Healthy piglets were used as the control. In the AAD-induced piglets, the lamina propria was edematous in the gyri centripetales. Piglets treated with ERFX were also edematous in gyri centrifugales. These edematous lamina propria contained larger amounts of inflammatory cells than observed in control tissues. ERFX-treated piglets had a more shallow crypt than PL-treated and control piglets. The mucosal tissue of the large intestine was more seriously damaged in the ERFX- than in the PL-treated piglets, which might have been caused by the high succinate and low SCFAs concentration in the digesta.     

壳聚糖对仔猪腹泻率的影响

本试验探讨不同的壳聚糖水平对仔猪腹泻率的影响,为壳聚糖在仔猪日粮中的科学应用提供理论依据。试验选用（60&#177;2）日龄雄性健康大白&#215;长白&#215;Ⅵ系杂种仔猪96头,分为4个处理,每个处理3个重复。4种日粮是在基础日粮中分别添加0、0．01％、0．05％和0．10％的壳聚糖配制而成。日粮壳聚糖对仔猪腹泻率的影响与添加剂量有关,添加0．01％和0．05％的壳聚糖对腹泻率的抑制作用效果较好。