Establishment and Application of a Multiplex RT-PCR Assay for Detection of PEDV,TGEV and PRoV

In this study,a multiplex RT-PCR assay was established to differentially detect porcine epidemic diarrhea virus (PEDV),porcine transmissible gastroenteritis virus (TGEV) and porcine rotavirus (PRoV) after optimization of the reaction conditions.Three pairs of primers PEDV-N,TGEV-M and PRoV-VP6 were designed for specifically amplifying PEDV N gene,TGEV M gene and PRoV VP6 gene,respectively.The assay could specifically amplify PEDV,TGEV and PRoV,but not classical swine fever virus (CSFV),porcine foot and mouth disease virus (FMDV),pseudorabies virus (PRV),porcine parvovirus (PPV) and porcine circovirus type 2 (PCV2).The detection limits of PEDV,TGEV and PRoV standard recombinant plasmids were 1.41×10³,1.41×10² and 1.41×10³ copies/μL,respectively.The repeated reaction under the same conditions obtained uniform results.The assay was used to detect a total number of 190 clinical samples,of which 42 (22.11%) samples were positive for PEDV,58 (30.53%) samples for TGEV and 34 (17.89%) samples for PRoV,and there were mixed infection among these viruses.The results indicated that this multiplex RT-PCR assay had the advantages of sensitivity,specificity and repeatability and provided a useful tool for differential detection and epidemiological investigation of PEDV,TGEV and PRoV.     

溶菌酶对犊牛腹泻和生长性能的影响

[目的]本试验研究溶菌酶在使用量不同情况下,对犊牛腹泻和生长性能的影响。[方法]选择无病、健康、体型近似的犊牛100 头,随机分为4 组,每组25 头,分别为对照组、低剂量组（Ⅰ组、1 g/头·天）、中剂量组（Ⅱ组、3 g/头·天）、高剂量组（Ⅲ组、5 g/头·天）,试验期30 天。在试验开始和结束时,观察犊牛粪便、腹泻、体重、体尺变化,并记录。[结果]试验期溶菌酶对犊牛的粪便评分有影响,1~30 天的粪便评分Ⅲ组显著低于对照组（P<0.05）,且Ⅲ组比对照组降低12.5%;1~15 天时腹泻率差异显著（P<0.05）,Ⅰ组、Ⅱ组、Ⅲ组与对照组相比分别降低23.02%、41.81%、53.64%,表明溶菌酶使用量多,腹泻效果更加明显;添加溶菌酶的Ⅱ组和Ⅲ组犊牛的平均日增重均显著高于对照组（P<0.05）,分别提高9.6%、12.0%;Ⅲ组胸围显著高于对照组,高3.0%。[结论]溶菌酶使用到合适剂量,不仅能降低犊牛腹泻率,还能促进犊牛的生长性能。     

Comparison of 2 methods for establishing rat model of chronic osmotic diarrhea

AIM: To compare 2 different rat models of chronic osmotic diarrhea with the lactose enrichment and magnesium citrate methods. METHODS: The SD rats (n=31) were randomly divided into lactose enrichment diarrhea group (Lac group, 12 rats), magnesium citrate diarrhea group (Mg group, 12 rats) and normal control group (Con group, 7 rats). The rats in Lac group were fed with a lactose enrichment diet and freely drank water. The rats in Mg group were fed with a standard diet and drank magnesium citrate solution. The Con group was fed with a standard diet and normal water. The general conditions and the characteristics of stool were observed every day. The grip strength and motor activity of the rats were measured after diarrhea for 14 d. The trans-epithelial resistance of the colonic mucosa was detected by Ussing chamber. The pathological changes of villus height and villus surface area in the intestine were observed with hemato-xylin-eosin staining. RESULTS: The rats in both diarrhea groups displayed the clinical manifestations of slow increase in body weight, spare and dull hair, lacking in strength and physical inactivity. The grip strength and motor activity of the rats in both diarrhea groups decreased significantly compared with Con group. The diarrhea rate and diarrhea index of the rats in the 2 diarrhea groups were both significantly higher than those in Con group (P<0.05). Moreover, the diarrhea index in Lac group was significantly increased compared with Mg group. The thickness of intestinal mucosa in Lac group was thinner than that in Mg group at the same time. However, the damage of intestinal villi in Mg group was more serious than that in Lac group.CONCLUSION: Both methods of lactose enrichment-induced diarrhea and magnesium citrate-induced diarrhea meet the requirement of chronic osmotic diarrhea, but there are different advantages between the diarrhea models. Therefore, we should select the correct diarrhea model to meet different aims of the experiments.     

水通道蛋白在调节肠道健康中的作用

水通道蛋白是一类可以高效选择性转运水分子的细胞膜通道蛋白,广泛分布于人和哺乳动物的胃肠道组织中,在机体液体转运和消化生理中发挥重要作用。本文总结了水通道蛋白的结构、分类及其在人、鼠类和仔猪胃肠道系统的表达和分布情况,并阐述了水通道蛋白与腹泻、肠上皮细胞迁移与修复、肠道屏障功能和肠道炎症相关疾病等过程的关系及其作用。     

女贞枸杞散预防断奶仔猪腹泻效果和对血液白细胞数的影响

为探讨以女贞子、枸杞子、穿心莲、大青叶等多味中药组成的"女贞枸杞散"对断奶仔猪腹泻的预防效果和对外周血各类白细胞数量的影响。根据体重相近、公母各半的原则选用80头体重为10kg左右的"杜长大"三元杂交断奶仔猪,随机分4组,分别为试验Ⅰ组(日粮中加0.4%"女贞枸杞散"),Ⅱ组(添加0.6%),Ⅲ组(添加0.8%)和对照组(普通日粮)。详细记录仔猪腹泻情况,计算腹泻率、腹泻频率、腹泻指数等。试验结束后采血,测定各类白细胞数量,结果表明,"女贞枸杞散"能有效地降低断奶仔猪的腹泻率,改善仔猪的生长性能,促进血液中的白细胞尤其是淋巴细胞的的生成,提高机体免疫力。     

新疆大蒜叶疫病的研究:Ⅱ病菌越冬致病性病害流行因素及防治

 试验结果表明,本菌(Stemphylium vesicarium(Wallr.) Simons)以病残体上分生孢子和菌丝体在田间和冷藏室越冬,9月至翌年6月检查,分生孢子萌发率为16.4~41.3%。人工接种试验指出,不论有无伤口,病菌均可侵染大蒜,但洋葱与大葱在伤口接种条件下病情较重。在自然条件下,大蒜的斑点型病状出现于6月下旬,7月为病情高峰期。影响病情的天气因素,主要是7月份温度和7月中旬的降水量(r=0.69)。如果7月雨次多、雨时长,病情即极为严重,可导致蒜田绝收。化学防治试验结果,以腐霉利防效最佳。从7月上旬开始,每间隔10~13天喷药1次。喷药2~3次即可取得较好的防治效果。     

Quantification of the transmission of classical swine fever virus between herds during the 1997-1998 epidemic in The Netherlands

In this study, we describe a method to quantify the transmission of Classical Swine Fever Virus (CSFV) between herds from data collected during the 1997–1998 epidemic in the Netherlands. From the contacts between infected herds and the serological findings shortly before depopulation, we estimated the week of virus introduction and the length of the period over which the herd emitted virus for each CSFV-infected herd. From these data, we estimated the infection-rate parameter β (the average number of herds infected by one infectious herd during one week) and the herd reproduction ratio, R_h (the average total number of secondary outbreaks caused by one infectious herd, i.e. in its entire infectious period), using a SIR-model for different sets of CSF control measures. When R_h > 1, an epidemic continues to grow. On the other hand, when R_h < 1 an epidemic will fade out.

During the phase before the first outbreak was diagnosed and no specific measures had been implemented, β was estimated at 1.09 and R_h at 6.8. In the subsequent phase infected herds were depopulated, movement restrictions were implemented, infected herds were traced forward and backward and the herds in the protection and surveillance zones were clinically inspected by the veterinary authorities (regional screening). This set of measures significantly reduced β to 0.38. However, R_h was 1.3 and thus still >1. Consequently, the number of outbreaks continued to grow. After a number of additional measures were implemented, the value of R_h was reduced to 0.5 and the epidemic came to an end. These measures included pre-emptive slaughter of herds that had been in contact with infected herds or were located near an infected herd, increased hygienic procedures, replacement of transports of pigs for welfare reasons by killing of young piglets and a breeding ban, and regional screening for CSF-infected herds by local veterinary practitioners.     

低聚木糖和酸化剂对断奶仔猪生产性能的影响

本试验研究了低聚木糖及其与酸化剂合用对断奶仔猪生产性能的影响。选择72头35±1日龄、平均断奶体重(10.33±0.34)kg的健康三元(杜×长×大)杂交仔猪,按体重和性别分成4个处理组,每处理组3个重复。第1组为对照组,饲喂玉米-豆粕型基础日粮;第2组为低聚木糖(200mg/kg)组;第3组为酸化剂(8g/kg)组;第4组为合用组,添加低聚木糖(200mg/kg)和酸化剂(8g/kg)。研究结果表明,各试验组日增重、料重比得到显著改善(P<0.05),腹泻率显著下降(P<0.01)。     

牛病毒性腹泻病毒持续感染的免疫学研究进展

牛病毒性腹泻病毒(BVDV)是危害养牛业的重要病原之一。BVDV感染能够引起广泛的临床症状,包括牛病毒性腹泻-黏膜病、持续感染、免疫耐受、繁殖障碍、血小板减少与出血综合征等。由于其病原的复杂性,给该病的控制及疫苗研制带来了一定的困难。近年来,对其免疫学的研究取得了一定进展,特别是在病原的基本免疫学特征、体液免疫、细胞免疫、免疫耐受和免疫调节方面做了深入研究,同时也在参与免疫应答的细胞因子方面做了研究,这些都为牛病毒性腹泻-黏膜病的预防和控制研究提供了新的思路。