Immune responses and enhanced disease resistance in Chinese drum, Miichthys miiuy (Basilewsky), after oral administration of live or dead cells of Clostridium butyrium CB2

Clostridium butyrium CB2 isolated from chickens was tested as a potential fish probiotic in the Chinese drum, Miichthys miiuy . Fish were fed live (CB), dead CB2 (D-CB) cells (10⁸ cells g⁻¹) or spent culture supernatant (SCS), for 30 days and challenged with Vibrio anguillarum or Aeromonas hydrophila . Survival was higher in both the CB and the CB-D fed groups, but the SCS group was not significantly different from the control. After feeding live or dead CB2 cells, there was increase in phagocytic activity of the head kidney macrophages, the lysozyme activity of serum and gut mucosa and immunoglobulin (Ig) level. The SCS group showed no obvious change in immune parameters. The results suggest that live or dead Clostridium butyrium CB2 has an immunomodulatory effect on fish.     

半胱胺对中华鳖生长性能和非特异性免疫功能的影响研究

试验研究了半胱胺对中华鳖生长性能和非特异性免疫功能的影响,并对其机理进行了初步探讨。选取750只健康、活泼的中华鳖,平均体重(220±10)g,分成5个组,每个组设3个重复,每个重复50只。对照组基础日粮中不添加半胱胺,其余各组为试验组,半胱胺添加方式及添加量如下:试验Ⅰ组断续添加(每6天添加一次,6000mg/kg);试验Ⅱ组、试验Ⅲ组、试验Ⅳ组分别每天连续添加500、1000和1500mg/kg。试验期90d。试验结果表明:半胱胺能够提高中华鳖成活率、增重率、摄食率和饲料效率,且连续添加效果好于断续添加,当半胱胺含量达到1000mg/kg时,中华鳖增重率、摄食率和饲料效率达到最高(P<0.05);半胱胺使中华鳖血清胃泌素水平升高,血清生长抑素水平降低;1000mg/kg半胱胺可以显著提高中华鳖肌肉RNA/DNA比率(P<0.05);半胱胺还可以增强中华鳖血清谷胱甘肽过氧化物酶、超氧化物歧化酶活性,但不同添加方式及剂量之间无显著性差异,血清溶菌酶含量在500mg/kg半胱胺添加组最高(P<0.05)。由本试验结果可以看出,半胱胺对中华鳖具有促进生长、增强免疫的作用,是一种良好的生物添加剂,且其适宜添加量为每天连续添加1000mg/kg。     

裙带菜凝集素对鲫的免疫诱导

将从裙带菜Undaria pinnatifida中提取的凝集素作为免疫诱导制剂,注射到鲫Carassius auratusw体内,在连续注射7 d和9 d时,分别对鲫血清中的溶菌酶、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的酶活性进行测定,结果表明,注射凝集素7 d和9 d时,鲫血清中溶菌酶的酶活性比对照组分别提高18.83%、12.69%,SOD的酶活性比对照组分别提高66.67%、139.99%,CAT酶活性比对照组分别提高16.60%、63.63%。说明凝集素可以明显提高鲫血清中这3种酶的表达水平。     

犬细小病毒核酸疫苗制备及免疫试验

以犬细小病毒基因组DNA为模板,应用PCR方法扩增了全长VP1基因,PCR产物经纯化和NotⅠ/BamHⅠ双酶切后与同样处理的真核表达载体pIRES进行连接,转化到感受态细胞JM109中,筛选了真核重组表达质粒pIRESVP1,并进行了序列测定。对6只9月龄犬分别接种不同剂量的pIRESVP1或空载体质粒及生理盐水,8周接种1次,每次接种前采血,测定血清的血凝抑制抗体效价,第2次免疫后即可检测到较高的血清效价。在第3次接种4周后,对全部实验犬进行攻毒,攻毒后第7天及第14天时采血,测定血清的血凝抑制抗体效价。所有接种pIRESVP1疫苗的犬均能抵抗CPV强毒的攻击,而接种空载体质粒和生理盐水的对照犬则全部发病。证明所构建的核酸疫苗(pIRESVP1)能使犬免受犬细小病毒强毒的感染。     

非洲猪瘟形势下猪流行性腹泻的防控措施

自2018年第一例非洲猪瘟疫情报道至今已经两年多了.为了快速扩繁猪群,许多清空场及新建场不得不大量外购来源复杂后备猪进群.因此,猪场在对猪流行性腹泻的防控上面临着更大的挑战.文章介绍了病毒性腹泻暴发的原因、防控难点和综合防控措施,期望为养殖业同人在实际生产中,科学防控猪流行性腹泻提供一定的参考借鉴.     

鹿源牛病毒性腹泻病毒核酸疫苗免疫实验研究

试验应用间接ELISA方法检测鹿源牛病毒性腹泻分离株CCSYD核酸疫苗体液免疫效果,应用淋巴细胞转化实验检测其细胞免疫效果,并与鹿源牛病毒性腹泻分离株CCSYD灭活苗和标准株C24V灭活苗进行比较。结果表明:鹿源牛病毒性腹泻分离株CCSYD核酸疫苗产生了较好的体液和细胞免疫应答,免疫前后免疫应答水平差异均显著(P<0.05)。     

斑马鱼幼鱼嗜中性粒细胞对副溶血弧菌清除的动态变化

副溶血弧菌(Vibrio parahaemolyticus)是人-兽-鱼共患的致病菌,严重威胁食品安全和人类的健康。为了研究宿主嗜中性粒细胞清除细菌感染的动态变化过程,利用红色荧光蛋白标记的副溶血弧菌Vp57~(RFP)菌株感染受精后48 h(hours post fertilization,hpf)的斑马鱼(Danio rerio)幼鱼耳泡,建立局部感染模型,并以大肠杆菌(Escherichia coli)Ec01菌株为对照,系统比较了副溶血弧菌和大肠杆菌的半数致死剂量(median lethal dose,LD_(50))、存活曲线以及和嗜中性粒细胞相互作用的动态过程,RT-qPCR验证炎症相关基因il1b和il10在感染前后的表达量变化。结果显示副溶血弧菌和大肠杆菌感染斑马鱼幼鱼的LD_(50)分别为7.90×10~8 CFU/mL和1.14×10~(11) CFU/mL,与感染大肠杆菌相比,斑马鱼感染副溶血弧菌后招募嗜中性粒细胞的速度更快,且数量更多。RT-qPCR结果发现斑马鱼感染副溶血弧菌后在相同时间点能够激活更高的il1b和il10基因的表达,引起强烈的炎症反应。基于以上研究,我们建立的副溶血弧菌-斑马鱼幼鱼耳泡局部感染模型,为进一步研究嗜中性粒细胞清除副溶血弧菌感染的动态过程,以及深入揭示天然免疫应答机制提供了依据。     

Marine-Derived Chitosan Nanoparticles Improved the Intestinal Histo-Morphometrical Features in Association with the Health and Immune Response of Grey Mullet (Liza ramada)

Marine-derived substances are known for their beneficial influences on aquatic animals’ performances and are recommended to improve intestinal health, immunity, and anti-oxidative status. The present study investigates the role of chitosan nanoparticles on the intestinal histo-morphometrical features in association with the health and immune response of Grey Mullet (Liza ramada). Chitosan nanoparticles are included in the diets at 0, 0.5, 1, and 2 g/kg and introduced to fish in a successive feeding trial for eight weeks. The final body weight (FBW), weight gain (WG), and specific growth rate (SGR) parameters are significantly increased while feed conversion ratio (FCR) decreases by chitosan nanoparticles compared to the control (p < 0.05). The morphometric analysis of the intestines reveals a significant improvement in villus height, villus width, and the number of goblet cells in chitosan-treated groups in a dose-dependent manner. Additionally, there is a positive correlation between the thickness of the enterocyte brush border and the chitosan dose, referring to an increasing absorptive activity. Histologically, the intestinal wall of Grey Mullet consists of four layers; mucosa, sub-mucosa, tunica muscularis (muscular layers), and serosa. The histological examination of the L. ramada intestine shows a normal histo-morphology. The epithelial layer of intestinal mucosa is thrown into elongated finger-like projections, the intestinal villi. The values of hemoglobin, hematocrit, red blood cells (RBCs), total protein (TP), albumin, and globulin are significantly increased in fish fed 1, and 2 g/kg of chitosan nanoparticles compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest levels of TP and albumin are observed in fish fed 1 g/kg diet (p < 0.05). The lysozyme activity and phagocytic index are significantly enhanced by feeding chitosan nanoparticles at 0.5, 1, and 2 g/kg, whereas the phagocytic activity is improved in fish fed 1 and 2 g/kg (p < 0.05). The highest lysozyme activity and phagocytic index are observed in fish fed 1 g/kg. SOD is significantly activated by feeding chitosan nanoparticles at 1 g/kg. Simultaneously, glutathione peroxidase (GPx) and catalase (CAT) activities also are enhanced by feeding chitosan at 1 and 2 g/kg, compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest GPx and CAT activities are observed in fish fed 1 g/kg (p < 0.05). Conversely, the malondialdehyde (MDA) levels are decreased by feeding chitosan at 1 and 2 g/kg, with the lowest being in fish fed 1 g/kg (p < 0.05). To summarize, the results elucidate that L. ramada fed dietary chitosan nanoparticles have a marked growth rate, immune response, and anti-oxidative response. These improvements are attributed to the potential role of chitosan nanoparticles in enhancing intestinal histo-morphometry and intestinal health. These results soundly support the possibility of using chitosan nanoparticles at 1–2 g/kg as a feasible functional supplement for aquatic animals.     

药用真菌发酵产物调控肉鸡内分泌及免疫力的研究

将药用真菌接种到以山芋渣为主并添加不同量的黄芪固态基质中进行发酵,获得发酵菌质,研究菌质对肉鸡内源性激素代谢、免疫力的影响。结果表明：菌质作为饲料添加剂饲喂AA肉鸡,对AA肉鸡的内源性激素有明显的调节作用,各试验组的生长激素浓度显著高于对照组（P〈0．05,P〈0．01）;试验1组的胰岛素、三碘甲腺原氨酸浓度显著高于对照组（P〈0．05）;试验2、3组的四碘甲腺原氨酸浓度显著高于对照组（P〈0．05,P〈0．01）;各试验组的皮质醇浓度低于对照组,但差异不显著。各试验组在配合疫苗使用的情况下,肉鸡血清中禽流感-H5亚型、新城疫抗体滴度显著高于对照组（P〈0．05,P〈0．001）;各试验组的免疫器官重量也比对照组高,说明菌质可提高肉鸡的免疫力。