牛病毒性腹泻-粘膜病病毒TaqMan荧光定量RT-PCR检测方法的建立及初步应用

为建立-种牛病毒性腹泻-粘膜病病毒（BVDV）的快速检测方法,根据GenBank中登录的BVDV基因序列,针对5’UTR的保守序列,设计合成了一对特异性引物和一条TaqMan荧光探针。通过对反应条件和反应体系进行优化,建立了一种能快速定量检测BVDV的荧光定量RT—PCR检测方法。通过对20份胎牛血清样品和猪瘟细胞苗半成品进行检测,对该方法的特异性、敏感性和重复性进行试验。结果显示,建立的方法能检测到BVDV,而对CSFV、PRRSV和MDBK细胞的扩增结果均为阴性,具有高度的特异性。在所检测的20份样品中,BVDV阳性6份（30％）。对所构建的标准品进行检测,在10^2-10^8拷贝／μL的范围内可以得到良好的动力学曲线,能检测低至10^3-10^4拷贝／mL的病毒量。表明所建立的检测方法具有快速、特异、灵敏、重复性好等特点,可用于临床及科研中对BVDV的快速定量检测和对牛血清及猪瘟兔化弱毒疫苗等生物制品中是否污染BVDV进行监测。     

禁抗后仔猪细菌性腹泻的临床诊断与防治

为了促进食品安全和养猪业的健康发展，禁止在饲料中使用促生长抗生素势在必行。随着我国的饲料禁抗相关法规的推行，饲料企业被严格禁止在饲料生产过程中加入抗生素，这也导致了猪场疾病尤其是肠道疾病的高发，而仔猪肠道发育未完全，免疫系统未发育成熟，更容易发生严重的腹泻甚至死亡。文章对几种常见的仔猪细菌性腹泻疾病进行了回顾，病原包括大肠杆菌、沙门氏菌、魏氏梭菌，讨论了它们的临床症状与防控措施，为猪场在防控这些细菌性腹泻提供参考。     

G and P genotype profiles of rotavirus A field strains circulating in beef and dairy cattle herds in Brazil, 2006–2015

The aim of this retrospective study was to use RT-PCR and nucleotide sequencing analysis to determine the G (VP7 gene) and P (VP4 gene) genotypes of 155 Brazilian bovine rotavirus A (RVA) wild-type strains detected in diarrheic calves from all Brazilian geographical regions from 2006 to 2015. The RVA strains evaluated belonged to the G6, G10, P[5], and P[11] genotypes. The G6P[5] genotype was prevalent (65.5%; P < 0.05) in beef, and the G10P[11] (38.4%) and G6P[11] (30.8%) genotypes were more prevalent in dairy cattle herds. The Midwest was the region with the highest number of genotyped RVA strains, where the genotypes G6, P[5], and P[11] were identified. Genotype combination G6-IV/P[5]-IX, prevalent in beef herds, and G6-III/P[11]-III or G10-IV/P[11]-III, prevalent in dairy herds, were detected. In addition, for the first time in Brazil, we detected the P[5] and P[11] genotype RVA strains that belong to lineage II and VII, respectively.     

一株猪流行性腹泻病毒的分离鉴定和遗传进化分析

从陕西地区26个疑似猪流行性腹泻病毒感染的猪场,采集发病仔猪的肛门拭子、粪便以及死亡仔猪的小肠,将其接种于Vero细胞,在添加胰酶的情况下,连续传代培养并观察,直至细胞出现典型的合胞体病变,经RT-PCR检测、测序、基因比对分析与动物回归实验,最终确定分离到一株PEDV病毒强毒株。经过测定毒株的S基因序列,进行序列比对分析和遗传进化分析,发现本次分离到的毒株与疫苗株CV777同源性较低,进化分支位于目前流行的PEDV G2a亚群。     

Molecular screening and risk factors of enterotoxigenic Escherichia coli and Salmonella spp. in diarrheic neonatal calves in Egypt

The aim of the present study was to carry out molecular epidemiological investigation on enterotoxigenic Escherichia coli (ETEC) K99 and Salmonella spp. in diarrheic neonatal calves. Fecal samples were obtained from 220 diarrheic calves at 9 farms related to four governorates in central and northern Egypt. E. coli and Salmonella spp. isolates were examined for E. coli K99 and Salmonella spp. using PCR. ETEC K99 was recovered from 20 (10.36 %) out of 193 isolates, whereas Salmonella spp. was recovered from nine calves (4.09%).Multivariable logistic regression was used to evaluate the risk factors associated with both infections. ETEC K99 was significantly affected by age (P < 0.01; OR: 1.812; CI 95%: 0.566–1.769), colostrum feeding practice (P < 0.01; OR: 5.525; CI 95%: 2.025–15.076), rotavirus infection (P < 0.001; OR: 2.220; CI 95%: 0.273–1.251), vaccination of pregnant dams with combined vaccine against rotavirus, coronavirus and E. coli (K99) (P < 0.001; OR: 4.753; CI 95%: 2.124–10.641), and vitamin E and selenium administration to the pregnant dam (P < 0.01; OR: 3.933; CI 95%: 0.703–1.248).Infection with Salmonella spp. was found to be significantly affected by the animal age (P < 0.05; OR: 0.376; CI 95%: 0.511–1.369), Hygiene (P < 0.05; OR: 0.628; CI 95%: 1.729–5.612), and region (P < 0. 01; OR: 0.970; CI 95%: 0.841–1.624).The results of the present study indicate the importance of PCR as rapid, effective and reliable tool for screening of ETEC and Salmonella spp. when confronted with cases of undifferentiated calf diarrhea. Moreover, identification of the risk factors associated with the spreading of bacteria causing diarrhea may be helpful for construction of suitable methods for prevention and control.     

Di-tri-octahedral smectite for the prevention of post-operative diarrhea in equids with surgical disease of the large intestine: Results of a randomized clinical trial

The aim of this study was to evaluate the effects of a commercially available di-tri-octahedral (DTO) smectite product on clinical signs and prevalence of post-operative diarrhea in horses with colic associated with disease of the large intestine. Sixty-seven horses with surgical disease of the large intestine were randomly assigned to be treated with DTO smectite (n = 37; 0.5 kg via nasogastric intubation every 24 h for 3 days post-operatively) or a placebo (n = 30). The effect of treatment on fecal scores and clinical and hematological parameters, including heart rate, mucous membrane color, temperature, total white blood cell count, total neutrophil count and total plasma protein values, were determined.Horses treated with DTO smectite had a significant reduction in the prevalence of post-operative diarrhea (10.8%), compared with controls (41.4%). A significant improvement in mucous membrane color was observed 72 h post-operatively in horses receiving treatment, compared with placebo. Administration of DTO smectite to colic patients with disease of the large intestine reduced the occurrence of diarrhea in the early post-operative period.     

Dietary supplementation with Acanthopanax senticosus extract enhances gut health in weanling piglets

The present study was conducted to investigate the effects of Acanthopanax senticosus extract (ASE) as a dietary additive on gut health in weanling piglets by examining diarrhea frequency, intestinal microbiota and morphology. A total of 96 Duroc× (Landrace × Yorkshire) piglets weaned at 21 days of age with an average initial body weight (BW) of 5.6 ± 0.4 kg were randomly assigned to 3 treatment groups with 4 duplicates of 8 piglets each. The piglets were fed basal diet to which had been added 0 or 1 g/kg of ASE, or 0.7 g/kg antibiotics, respectively. Fecal consistence was monitored twice daily and the frequency of diarrhea was calculated. On day 21 after the initiation of supplementation, 8 piglets were randomly selected from each treatment group (2 piglets per pen) and slaughtered. The jejunum, ileum, colon and cecum were then excised and fixed in 10% neutral formalin solution to determine villus height and crypt depth, after their contents were collected to determine microbiota. The results showed that dietary supplementation with ASE increased (P < 0.05) the density of bacterial populations that co-migrated with Lactobacillus amylovorus, Lactobacillus salivarius, Bacillus subtilis, and Clostridium lituseburens, but decreased (P < 0.05) those co-migrating with Staphylococcus aureus, Salmonella typhimurium, Ruminococcus forques, and E. coli O157:H7 in the PCR-DGGE profiling analysis when compared with the control group. The villus height of the duodenum, jejunum and ileum increased (P < 0.05) by 14.8, 13.7 and 10.0%, while the crypt depth decreased (P < 0.05) by 17.9, 9.1 and 12.1%, respectively, in response to dietary ASE supplementation. Additionally dietary supplementation with ASE or an antibiotic decreased (P < 0.05) the frequency of diarrhea by 55.6 and 52.2%, respectively, compared with the control group. In conclusion, these findings suggest that dietary supplementation with ASE could regulate the microbiota composition and maintain a normal morphology of gut mucosa in weanling piglets, thereby decreasing diarrhea that resulted from weaning stress.     

益生菌·低聚木糖制剂对仔猪生长性能和黄白痢的预防效果

选用2胎龄斯格母猪20头,随机分成5个处理,每处理4次重复,仔猪出生3 d内吃乳前饲喂1 mL液态微生态制剂,诱食后在基础日粮中分别添加0.1% 微胶囊(Ⅰ)、0.05% 低聚木糖(Ⅱ)、0.1% 微胶囊和0.05% 低聚木糖(Ⅲ)、0.1% 菌粉和0.05% 低聚木糖(Ⅳ)及对照组(Ⅴ),观察其对仔猪生长性能、黄白痢发生率和死亡率的影响.结果表明:7日龄内饲喂菌液对体质量增加无明显影响(P>0.05),黄白痢发生率与对照组差异不显著,但仔猪死亡率明显低于对照组(P<0.01).8～28日龄Ⅲ组与Ⅰ、Ⅱ、Ⅴ组仔猪体质量增加差异显著(P<0.05),所有试验组与对照组黄白痢发生率差异极显著(P<0.01),Ⅲ、Ⅳ组与Ⅰ、Ⅱ组黄白痢发生率差异极显著(P<0.01);29～40日龄Ⅲ组与Ⅰ、Ⅱ、Ⅳ、Ⅴ组仔猪体质量增加差异显著(P<0.05),Ⅲ组仔猪黄白痢发生率与其他试验组和对照组差异极显著(P<0.01);试验Ⅰ、Ⅱ、Ⅳ组与Ⅴ组差异显著(P<0.05).     

仔猪腹泻的病因分析及综合防制

仔猪腹泻是引起仔猪死亡的重要原因之一。该病病因复杂,始终是困扰养猪业的一大难题。文章详细分析了引起仔猪腹泻的非传染性病因（环境、仔猪自身等）及传染性病因（细菌、病毒、寄生虫等）,提出了预防为主,加强饲养管理及配合药物治疗的综合防制方案,对减少仔猪腹泻,维护养猪业的健康发展具有一定的指导意义。