牦牛源正呼肠孤病毒2型的检测和分离鉴定

旨在调查川西北牦牛哺乳动物正呼肠孤病毒（MRV）的感染情况并分离病毒。采用RT-PCR方法，对采自川西北15个牧场的72份牦牛腹泻粪便样本和其中5个牧场的15份腹泻牦牛血清样本进行MRV检测，阳性样本进一步用分型PCR确定其血清型。结果显示，粪便样本中MRV检出率为20.83%（15/72），血清2型的比例为60%（9/15）；血清样本中MRV检出率为40%（6/15），血清2型的比例为83.33%（5/6）；未检测到其他血清型。成功地从腹泻粪便中分离到1株MRV血清2型毒株（TCID₅₀为4×10^-8.56·mL^-1），并获得长度为23 587 bp的分离株全基因组，该分离株与中国猪源毒株的遗传关系最近；与GenBank中所有的MRV S1基因相比，该分离株有4个独特的氨基酸突变。本研究从牦牛中检测到MRV，并分离到1株牛源MRV血清2型毒株，为进一步研究MRV血清2型生物学特性奠定了基础。     

The effects of density and artificial substrate on intensive shrimp Litopenaeus vannamei nursery production

Recirculating aquaculture systems (RAS) can be installed indoors, allowing year-round production of tropical animals in nearly any climate. A nursery phase is commonly used in Litopenaeus vannamei production since it allows for enhanced biosecurity and better quantification of animals while reducing space requirements. However, it is unclear whether animal density and inclusion of artificial substrate may improve shrimp performance during the nursery phase. In this experiment, we compared shrimp production parameters in two stocking densities with or without the use of an artificial substrate by creating four treatments: low-density LD; 1500 PL/m⁻³, low-density with substrate LDS, high-density HD; 3000 PL/m⁻³), and high-density with substrate (HDS). The LDS and HDS treatments included 0.46-m² of high-density polyethylene 2.5-cm mesh as a substrate, which increased the tank surface area by 21 %. Each treatment was randomly assigned to four 160-l culture tanks, each with a biofilter. The shrimp had an initial weight of 4 mg and were grown for 50 days. The low-density treatments had significantly higher dissolved oxygen (DO) and pH than the high-density treatments (P ≤ 0.001). Specifically, LDS had the highest DO and pH followed by the LD, HD, and HDS treatments, respectively. High-density treatments had significantly higher NO₂-N levels than low-density treatments during week 2 of the experiment when an unusually high concentration of nitrite was observed. FCR was significantly lower in both low-density treatments than in high-density treatments. At harvest, the total biomass (kg m⁻³) was significantly higher in high-density treatments than in low-density treatments (P ≤ 0.001), and the HDS treatment had a significantly greater biomass output than HD. Producers should consider artificial substrate and higher densities during nursery production to maximize shrimp production; however, the effects on water quality should also be taken into account.     

牦牛瘤胃微生物抗生素抗性基因对3种外源性刺激因子的响应

通过给牦牛投喂硫酸头孢喹肟(CEF)、盐酸二氟沙星(DIF)和黄曲霉毒素B1(AFB1),并进行瘤胃微生物宏基因组测序,旨在揭示这3种外源性刺激因子对抗生素抗性基因(ARGs)种类、抗性类型、抗性机制等的影响,对于深入研究微生物抗性组特征和抗性机制具有重要价值。选取15头牦牛,随机分5组。Cef组和Dif组分别根据说明书推荐剂量按体重计算、灌服CEF 1 mg·kg^-1和DIF 1 mL·kg^-1;E1组和E2组分别按采食量投喂AFB120、60μg·kg^-1;C组为对照组。处理7 d后,采集瘤胃液,提取DNA,Illumina HiSeq测序,对reads counts进行标准化得到TPM值,并进行方差分析。结果显示,对照组共获得132种ARGs,分属30种抗性类型,其中,四环素类tetQ和tetW基因丰度较高;Cef组tetW基因丰度增加(P<0.05),Dif组tetQ丰度增加(P<0.05);Cef组四环素类和头孢菌素类抗性基因丰度增加(P<0.05),Dif组四环素类和氨基香豆素类抗性基因丰度增加(P<0.05),E1组氨基香豆素和青霉烯类抗性基因丰度增加(P<0.05),E2组青霉烯类、头孢菌素类等9类抗性基因丰度均增多(P<0.05);Dif组Erm基因23S核糖体RNA甲基转移酶丰度增加(P<0.05),E2组中ATP结合盒超家族等3种抗性机制相关基因的丰度增加(P<0.05);3种因子均显著增加四环素类ARGs宿主的种类。结论:瘤胃是蕴含丰富ARGs的储藏库,其中,四环素类抗生素抗性基因tetQ和tetW是主要的ARGs。不仅CEF和DIF使部分ARGs的种类、抗性类型以及耐药机制相关酶等的丰度升高,增加瘤胃微生物的耐药性,而且AFB1也具有类似作用,且高剂量AFB1对抗性类型的影响范围较抗生素大。这3种因子还导致携带四环素类ARGs宿主微生物的种类数量增加,从而强化横向转移机制,加快ARGs传播,增强微生物对四环素类的耐药性。     

新疆卡提拉玛工艺配方的研究

熟牛油、生奶皮、白砂糖是制作新疆塔吉克族特色面制品的主要原料。通过正交试验确定三者在卡提拉玛中的添加量，研制具有新疆独特风味的卡提拉玛。在基本配方（高筋小麦粉500 g，酵母3 g，食盐2 g，鸡蛋清30 g，牛奶185 mL，饮用水100 mL，清油4 mL）的基础上，确定三者最佳添加量（以高筋粉为基重）。结果表明：加入熟牛油6 mL、生奶皮15 g、白砂糖3 g，可制作出色泽金黄、口感醇香、质地酥脆、具有生奶皮特有香气的新疆卡提拉玛。     

A Lymphoid Organ Specific Anti-Lipopolysaccharide Factor from Litopenaeus vannamei Exhibits Strong Antimicrobial Activities

Different shrimp species are known to possess apparent distinct resistance to different pathogens in aquaculture. However, the molecular mechanism underlying this finding still remains unknown. One kind of important antimicrobial peptides, anti-lipopolysaccharide factors (ALF), exhibit broad-spectrum antimicrobial activities. Here, we reported a newly identified ALF from the shrimp Litopenaeus vannamei and compared the immune function with its counterpart in the shrimp Fenneropenaeus chinensis. The ALF, designated as LvALF8, was specifically expressed in the lymphoid organ of L. vannamei. The expression level of LvALF8 was apparently changed after white spot syndrome virus (WSSV) or Vibrio parahaemolyticus challenges. The synthetic LBD peptide of LvALF8 (LvALF8-LBD) showed strong antibacterial activities against most tested Gram-negative and Gram-positive bacteria. LvALF8-LBD could also inhibit the in vivo propagation of WSSV similar as FcALF8-LBD, the LBD of LvALF8 counterpart in F. chinensis. However, LvALF8-LBD and FcALF8-LBD exhibited apparently different antibacterial activity against V. parahaemolyticus, the main pathogen causing acute hepatopancreatic necrosis disease (AHPND) of affected shrimp. A structural analysis showed that the positive net charge and amphipathicity characteristics of LvALF8-LBD peptide were speculated as two important components for its enhanced antimicrobial activity compared to those of FcALF8-LBD. These new findings may not only provide some evidence to explain the distinct disease resistance among different shrimp species, but also lay out new research ground for the testing and development of LBD-originated antimicrobial peptides to control of shrimp diseases.     

Comparing biofloc,clear-water,and hybrid nursery systems (Part I): Shrimp (Litopenaeus vannamei) production,water quality,and stable isotope dynamics

Indoor, intensive, nursery-based recirculating aquaculture systems (RAS) can provide high-quality juvenile shrimp for indoor or pond-based production systems in a biosecure manner. However, it is unclear what type of RAS is most appropriate for indoor shrimp nurseries. This study compared three types of RAS nurseries: biofloc (BF), clear-water (CW), and hybrid (HY). Each treatment included four, randomly assigned 160 L (0.35-m²) tanks that were stocked with 3000 post-larvae shrimp m⁻³. The post-larvae (PL10) shrimp had an initial average weight of 7 ± 0.0 mg and were grown for 48 days. The BF tanks included external settling chambers as the only filtration mechanism. The CW tanks had settling chambers, foam fractionators, and external biofilters to fully clarify the water and process nitrogenous waste. Hybrid tanks included settling chambers, and external biofilters to maintain some suspended solids along with external biofiltration. Overall, the CW treatment had significantly higher dissolved oxygen (DO) and pH levels than the BF and HY systems. The HY treatment had significantly higher DO than the BF treatment. Nitrite concentration was significantly higher in the HY treatment than the CW treatment. Turbidity in the BF treatment was significantly higher than the other treatments. On the final sample date, the BF treatment had significantly higher nitrite and nitrate concentrations than the other treatments. Differences between treatments in terms of shrimp survival, mean harvest weight, specific growth rate, and feed conversion ratio were not significant. The final weight of the shrimp at 48 days for the BF, CW, and HY were 670 mg, 640 mg, and 590 mg respectively. A stable isotope mixing model indicated that, in the BF treatment, 13% of the C and 34% of the N in harvested shrimp tissue may have originated from biofloc material, signifying some nutrient recycling. The nitrification process was more effective with the inclusion of an external biofilter. All three system types appear suitable for RAS shrimp nursery production although consideration should be given to water quality consistency and filtration costs.     

白茶寿眉预防小鼠肥胖作用研究及安全性评价

取1、3、7年寿眉(白茶)制备白茶提取物(White tea extracts,WTEs)。在对雄性C57BL/6J小鼠喂饲高脂饲料的同时给予WTEs干预,诱导建立预肥胖模型。观察小鼠体质量增量、摄食量、脂肪和肝脏组织病理学变化,统计脂肪湿重,测定血脂及血转氨酶水平。q PCR测定小鼠肝脏中脂代谢相关基因表达。综合评价WTEs预防小鼠肥胖功效及其安全性,并对其机制进行探究。结果显示,WTEs在无肝毒性和生长抑制作用条件下,能抑制脂肪细胞分化和脂肪的过度积累,还能下调SREBP-1c、FAS和ACC1的转录水平,抑制肝脏脂肪酸合成和脂肪过度积累,有效预防肥胖和脂肪肝的发生。高剂量下,WTEs预防肥胖功效随白茶年份的增加而显著降低。     

WSSV envelope protein VP51B links structural protein complexes and may mediate virus infection

White spot syndrome virus (WSSV), an enveloped double‐stranded DNA virus, is the causative agent of a disease that has led to severe mortalities of cultured shrimps in Taiwan and many other countries. In the previous study, Penaeus monodon chitin‐binding protein (CBP) and glucose transporter 1 (Glut1), two cell membrane proteins, were found to at least interact with other 10 WSSV envelope proteins including VP51B. These envelope proteins might form a protein complex. According to the known information, VP51B was used to identify its role in the protein complex. Western blotting of the intact viral particles and fractionation of the viral components confirmed that VP51B is one of WSSV envelope proteins. In this study, the protein–protein interaction between VP51B and other WSSV envelope proteins was identified by far‐western blot experiment and VP51B was found to interact with VP24, VP31, VP32, VP39B and VP41A. Furthermore, the in vivo neutralization experiment using recombinant VP51B plus with VP39B showed the best inhibition. These data indicate that VP51B participates in the WSSV protein complex and plays an important role in WSSV infection.     

Effect of dietary GroBiotic®‐A supplementation as a prebiotic on the intestinal microflora,growth performance,haemato‐serological parameters,survival rate and body composition in juvenile beluga (Huso huso Linnaeus, 1754)

The objectives of the present study were to investigate of GroBiotic^®‐A (GBA) on growth, autochthonous intestinal microbiota and haemato‐serological parameters of beluga juvenile. A total of 180 fish (40.82 ± 5.81 g) were fed diets containing graded levels of GBA (0, 5, 10 and 20 g kg^?1 diet) for 8 weeks. No significantly differences in body composition, total viable aerobic bacteria, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, neutrophils, eosinophils, monocytes, albumin, glucose, triglyceride, cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase were detected between treatment groups. The fish fed on 10 and 20 g kg^?1 GBA significantly showed higher lactic acid bacteria, final weight, body weight increases, weight gain and lower feed conversion ratio compared with the control and 5 g kg^?1 groups. The group fed 20 g kg^?1 GBA showed a highly significant difference in condition factor, survival rate, final length, total red and white blood cells, lymphocytes, haematocrit, haemoglobin, total protein, total immunoglobulin and lysozyme activity. The specific growth rate of the treatment groups was significantly elevated compared with the control groups. These results indicated that GBA at level 20 g kg^?1 improved growth, welfare and survival of beluga juvenile.     

四川石渠县牦牛源蜱感染巴尔通体的分子流行病学调查

为了解四川省石渠县牦牛体表寄生蜱的种类及其巴尔通体的感染情况。采集牦牛体表的蜱,经形态学初步鉴定后,提取蜱总DNA,PCR扩增蜱细胞色素氧化酶Ⅰ(COⅠ)基因和巴尔通体gltA基因,对阳性产物测序、比对,并构建系统进化树,从而确定蜱及其携带巴尔通体的种类。结果表明:在石渠县4个乡共采集到蜱818只,其中西藏革蜱占78.97%(646/818)、青海血蜱占21.03%(172/818);蜱巴尔通体的总感染率为30.07%,其中阿日扎乡、麻甲乡、德荣玛乡、长须干玛乡蜱巴尔通体的感染率分别为4.76%、76.79%、12.50%、17.95%,麻甲乡西藏革蜱巴尔通体感染率显著高于其他地点(P<0.01),麻甲乡青海血蜱巴尔通体感染率(79.07%)高于西藏革蜱(69.23%),但无显著差异(P>0.05)。经比对分析,总共得到3条序列(uncultured Bartonella sp. shiqu 1,uncultured Bartonella sp. shiqu 2和uncultured Bartonella sp. shiqu 3)。遗传进化分析显示,uncultured Bartonella sp. shiqu 1和uncultured Bartonella sp. shiqu 2与未定种的Bartonella spp. RF124HAIN (FJ464240)亲缘关系最近;uncultured Bartonella sp. Shiqu 3与对人有致病性的Bartonella melophagi亲缘关系最近。综上,石渠县存在西藏革蜱和青海血蜱,蜱巴尔通体感染率较高,并且具有感染人的风险。