鸡毒霉形体黏附素截短蛋白表达条件的优化

研究了培养温度、培养时间、培养基pH值以及异丙基硫代-β-D-半乳糖苷(isopropylthio-β-D-galacto-side,IPTG)浓度等不同条件对鸡毒霉形体黏附素截短蛋白在大肠杆菌中表达量的影响。经SDS-PAGE分析表明:诱导温度为28℃、诱导时间为9 h、培养基pH值为7.0、IPTG浓度为0.010 mmol/L时目的蛋白在细菌裂解沉淀和上清液中均最大量表达,分别达到30%和17%左右。上清液经GST.Bind Resin亲和层析分离纯化后,洗脱产物中pMGAⅣ融合蛋白纯度高达95%。Western-blot分析表明,该蛋白可被鸡毒霉形体阳性血清识别,具有较好的免疫学活性。     

日粮中鱼蛋白水解物对黄颡鱼生长、体成分和血清生理指标的影响

为了探讨鱼蛋白水解物对黄颡鱼生产性能的影响,以日本鳀粉为对照,以实用型黄颡鱼饲料配方模式为基础开展实验:1以30.5%鱼粉为对照(FM),在相同配方模式下,以6%鱼蛋白水解物(MPH6)替代20%的鱼粉;2以30.5%鱼粉为对照(FM),在无鱼粉日粮中分别添加3%(FPH3)、6%(FPH6)、12%(FPH12)鱼蛋白水解物;共设计5组等氮等能实验日粮,在池塘网箱中养殖黄颡鱼[初始体质量(30.08±0.35)g]60 d。结果显示:与FM相比,FPH12在SGR、FCR、PRR和FRR方面均无显著差异,而MPH6、FPH3、FPH6组SGR降低了15.45%~24.39%,FCR升高了32.14%~42.86%,MPH6、FPH6差异显著,在PRR和FRR方面,MPH6、FPH3、FPH6组PRR降低了21.11%~27.78%,MPH6组FRR降低了41.51%;全鱼水分、粗蛋白、粗脂肪和灰分各组间差异不显著,FPH3、FPH6、FPH12肌肉多种游离氨基酸水平显著高于FM,其中Thr、Val、His与其在日粮中的水平显著相关;FPH6组HSI显著低于FM,鱼蛋白水解物对CP、VSI、肠体比的影响不显著;血清AST、ALT、HDL、LDL、TP、CHOL、TG无显著差异,FPH3组ALB显著低于FM。研究表明:黄颡鱼日粮中,12%鱼蛋白水解物(干物质)与30.5%鱼粉在生长速度、饲料效率、血清生理指标等方面具有一定的等效性;过高的植物蛋白日粮影响了黄颡鱼的生产性能;饲喂鱼蛋白水解物日粮使黄颡鱼肌肉游离氨基酸的含量升高,特别是呈味氨基酸的含量增加。     

盐度胁迫对高体革鯻幼鱼消化酶活力的影响

研究了盐度对高体革（Scortum barcoo）幼鱼消化酶活力的影响。在温度25±2℃下,将平均体质量为2.50±0.23 g的高体革幼鱼分别饲养在盐度0,5,8,11,13和16条件下20 d,检测幼鱼胃蛋白酶、淀粉酶和脂肪酶活性的差异。结果表明：盐度胁迫对高体革幼鱼胃蛋白酶活力有显著影响（P〈0.05）,对脂肪酶和淀粉酶活力有极显著影响（P〈0.01）。随着盐度的升高,胃蛋白酶活力先升后降,当盐度升高到13时又急剧升高;脂肪酶的活力则是随盐度的升高而受到抑制,当盐度升高到13时也急剧升高,盐度16时最高;而淀粉酶活力变化则是盐度升高时显著降低。这说明环境盐度对高体革消化酶比活力的影响比较大,不同消化酶随盐度变化规律有所差异。     

两种盐度下凡纳滨对虾饲料中的最适动植物蛋白比

试验研究了不同盐度下（3和22）,以鱼粉和大豆浓缩蛋白为蛋白源,配制6种不同动植物蛋白比的饲料对凡纳滨对虾生长、成活和肝胰腺可溶性蛋白质含量的影响,饲养试验为期40 d。结果显示：（1）饲料动植物蛋白比可显著影响凡纳滨对虾增重率、成活率、肝体指数、肥满度和肝胰腺中可溶性蛋白质含量。增重率随饲料动植物蛋白比升高而升高,但当饲料中动植物蛋白比升至29∶8时,增重率不再明显升高,其它指标均先随饲料动植物蛋白比升高至一定程度,而后则稍有下降;（2）盐度22组对虾的增重率、成活率和肥满度显著高于盐度3组对虾,肝体指数却显著低于盐度3组,不同的盐度对凡纳滨对虾肝胰腺可溶性蛋白含量的影响不显著;（3）双因素方差分析结果显示,盐度和饲料动植物蛋白比对凡纳滨对虾增重率、成活率和肝体指数存在显著交互作用,最大值分别出现在盐度22下全动物蛋白饲料组、盐度22下全动物蛋白和动植物蛋白为29∶8的饲料组、盐度3下饲料动植物蛋白比为14∶23的饲料组中;（4）Broken-Line分析表明,3‰盐度下凡纳滨对虾最适饲料蛋白比为29.12∶7.79~30.29∶6.71,盐度22时为26.05∶10.95~29.03∶7.44。结果提示,饲料中氨基酸的组成和含量会随配方中动植物蛋白配比而改变,且不同盐度下凡纳滨对虾对饲料中动植物蛋白比的要求有所不同,但配饵中适当的动植物蛋白比可以满足虾对各种氨基酸的适宜需求。因此,在养殖过程中,需结合实际的养殖环境和饲料蛋白源种类,来设计适宜的实用饲料配方,这样才能达到降低生产成本,提高经济效益的目的。     

Relationship Between Dietary Protein Source and Feeding Frequency During Feeding Nile Tilapia,Oreochromis niloticus (L.) Cultured in Concrete Tanks

ABSTRACT

A 24-week feeding trial was conducted in concrete tanks with Nile tilapia, Oreochromis niloticus (L.) with an average initial weight and length of 50.87 ± 6.03 g and 14.4 ± 0.45 cm, respectively, to examine the effect of two protein sources and two feeding frequencies on growth performance, production traits, and body composition. Twelve 4 m³ concrete tanks (2 × 2 × 1.25 m, long, width, and height) were each stocked with 100 fish and fed diets containing either fish meal protein or soybean meal protein at two feeding frequency of two times daily or four times daily (2 × 2 × 3 factorial experiment). The results revealed that there was no significant effect of dietary protein sources on growth rate, whereas there was a significant increase in growth rate with increasing feeding frequency (P < 0.05). The same trend was also observed for mean body weight (g), production rate (kg/m3), specific growth rate (SGR % day^?1). The best final mean body weight (g), specific growth rate (SGR % day^?1), and production rate (kg/m3) were recorded in groups of fish fed with feeding frequency four times daily. Whole fish moisture, protein, fat, and ash contents were significantly influenced (P ≤ 0.05) by protein sources and feeding frequency, except protein not influenced with feeding frequency. Energy was significantly (P < 0.05) influenced by feeding frequency, but not by dietary protein sources. Economic evaluation indicated that soybean meal (Diet B) at four times feeding daily was the most cost-effective and affordable feed strategy for farmers. We conclude that a soybean diet fed four times daily is recommended for adult Nile tilapia reared in concrete tanks.     

基于反相高效液相色谱技术测定市售羊乳粉中蛋白

为鉴别及检测不同地区市售羊乳粉的品质,基于反相高效液相色谱(reversed-phase high performance liquid chromatography,RP-HPLC)技术,将35种市售羊乳粉根据陕西地区和非陕西地区、纯羊乳粉和配方羊乳粉进行分类与检测,研究不同产地、不同种类羊乳粉的差异,对羊乳粉品质进行分析判别。结果表明：对市售羊乳粉乳蛋白进行RP-HPLC测定发现,配方羊乳粉α_s2-酪蛋白(α_s2-casein,α_s2-CN)和β-CN相对含量分别低于纯羊乳粉20%和24%,κ-CN和乳清蛋白相对含量分别高于纯羊乳粉30%和50%;不同地区纯羊乳粉乳蛋白组成及含量相似。因此,通过RP-HPLC可以实现对市售羊乳粉中乳蛋白的测定和质量评价。     

复方黄芪颗粒防治鸡传染性法氏囊病的疗效试验

应用复方黄芪颗粒对鸡传染性法氏囊炎病毒人工感染病例进行试验,结果表明,有效率达到80%~92%,治愈率达到68%~76%,均高于对照药物,差异极显著(P<0.01)。推荐剂量为每升饮水中加入1g复方黄芪颗粒,连用5 d。     

坝竹和小篷竹景观效果及实生苗培育

实地调查了黔北坝竹和黔南小蓬竹天然林景观效果,并进行了引种及实生苗培育试验。结果表明：2个竹种在喀斯特石山地沿岩壁下垂如帘,形态飘逸,颇为美观。坝竹实生苗人工培育1年时,每株苗分蘖11个左右,苗高50 cm左右,培育15个月时苗高74 cm左右;小蓬竹实生苗人工培育1年时,每株苗分蘖7个左右,苗高72 cm左右,培育15个月时苗高77 cm左右。研究结果可为坝竹和小篷竹的边坡绿化及盆栽观赏提供参考。     

1株黄曲霉颉颃菌的鉴定及其抑菌活性成分分析

【目的】筛选黄曲霉颉颃菌,探究其最佳培养条件及有效抗菌成分,为实际生产中应用生物防治方法抑制黄曲霉污染提供新的理论依据。【方法】通过平板对峙法及分子生物学鉴定方法对待测菌株进行筛选鉴定。以抑菌率作为衡量标准,判断所得黄曲霉颉颃菌的最佳培养温度、培养时间、接种量、转速、pH及装液量。通过硫酸铵沉淀法从颉颃菌无菌发酵上清液中提取蛋白,观测粗提物对黄曲霉菌丝生长的影响。通过离子交换色谱和凝胶过滤色谱进一步纯化粗提蛋白,并进行质谱测序及序列对比分析,筛选鉴定颉颃菌产生的有效抑菌成分。【结果】筛选到1株解淀粉芽孢杆菌B10(菌种保藏号CCTCCNO:M2018353)。最佳培养条件为温度40℃,培养时间36 h,接种量3%,转速120 r/min, pH 6.0,装液量30 mL,最大抑菌率可达46.56%。其粗提蛋白能有效抑制黄曲霉生长,破坏菌丝正常形态,并从中筛选出几丁质结合蛋白、壳聚糖酶、葡聚糖酶等8种可能的有效抑菌成分。【结论】本试验分离鉴定出1株抑制黄曲霉正常生长发育的解淀粉芽孢杆菌B10,其产生的多种蛋白成分对黄曲霉有显著抑菌活性。     

Comparison of Physicochemical and Structural Properties of Acid-Soluble and Pepsin-Soluble Collagens from Blacktip Reef Shark Skin

Fish collagen has been widely used in tissue engineering (TE) applications as an implant, which is generally transplanted into target tissue with stem cells for better regeneration ability. In this case, the success rate of this research depends on the fundamental components of fish collagen such as amino acid composition, structural and rheological properties. Therefore, researchers have been trying to find an innovative raw material from marine origins for tissue engineering applications. Based on this concept, collagens such as acid-soluble (ASC) and pepsin-soluble (PSC) were extracted from a new type of cartilaginous fish, the blacktip reef shark, for the first time, and were further investigated for physicochemical, protein pattern, microstructural and peptide mapping. The study results confirmed that the extracted collagens resemble the protein pattern of type-I collagen comprising the α₁, α₂, β and γ chains. The hydrophobic amino acids were dominant in both collagens with glycine and hydroxyproline as major amino acids. From the FTIR spectra, α helix (27.72 and 26.32%), β-sheet (22.24 and 23.35%), β-turn (21.34 and 22.08%), triple helix (14.11 and 14.13%) and random coil (14.59 and 14.12%) structures of ASC and PSC were confirmed, respectively. Collagens retained their triple helical and secondary structure well. Both collagens had maximum solubility at 3% NaCl and pH 4, and had absorbance maxima at 234 nm, respectively. The peptide mapping was almost similar for ASC and PSC at pH 2, generating peptides ranging from 15 to 200 kDa, with 23 kDa as a major peptide fragment. The microstructural analysis confirmed the homogenous fibrillar nature of collagens with more interconnected networks. Overall, the preset study concluded that collagen can be extracted more efficiently without disturbing the secondary structure by pepsin treatment. Therefore, the blacktip reef shark skin could serve as a potential source for collagen extraction for the pharmaceutical and biomedical applications.