脱脂柞蚕蛹蛋白用复合中性蛋白酶水解制备多肽的工艺条件优化试验

利用蛋白酶对脱脂柞蚕蛹蛋白进行酶解制备具有活性的多肽,可以提高柞蚕蛹蛋白的应用价值。为了建立适合工业化应用的脱脂柞蚕蛹蛋白的酶解工艺条件,首先采用单因素试验考察料液质量浓度、复合中性蛋白酶种类与使用浓度、酶解温度和时间4个因素对脱脂柞蚕蛹蛋白水解度的影响,得出较适合的工艺条件是:料液质量浓度60 g/L,3 g/L复合中性蛋白酶中的风味蛋白酶与蚕蛹专用复配蛋白酶按1∶1质量比混合,酶解温度55℃,酶解时间10 h。进一步采用Box-Behnken设计及响应面分析法对工艺条件进行优化,在复合中性蛋白酶质量浓度4.85 g/L、料液质量浓度41 g/L、酶解温度55℃、酶解时间10.66 h的最佳酶解条件下,脱脂柞蚕蛹蛋白的理论水解度为45.82%,实际水解度为45.75%。利用优化的酶解工艺条件制备柞蚕蛹蛋白活性肽,具有酶解效率高、稳定性好、操作简单、生产成本低的特点。     

柞蚕幼虫在体脑神经节自发放电活动的初步研究

以柞蚕 5龄幼虫为材料 ,运用微电极电生理技术 ,以细胞外记录方法记录了柞蚕幼虫在体脑神经节的自发放电活动 ,并观察了胞外Na+、K+、Ca2 +的浓度变化对放电活动的影响。结果表明 ,柞蚕幼虫脑神经节的胞外放电活动有 3种形式 ,其离子机制可能既有Na+动作电位 ,又有Ca2 +动作电位。     

柞蚕对核型多角体经口感染的抵抗性研究

取不同素质的柞蚕,于1、2、5龄饷食期,用离心提纯的新鲜核型多角体病毒液以五倍系列稀释5—6六个浓度,分别进行经口定量添毒测定,然后根据各组发病情况,求出半数致死量(LD_(50)),来衡量各类型柞蚕的抗毒力大小。初步认为:(1)河南一化性柞蚕品种,对核型多角体病毒抵抗性差异在十倍左右。(2)同品种不同蛾系间的抗毒力也有一定差异。(3)辉点蚕的抗毒力是二对侧点蚕<一对侧点蚕<正常蚕。(4)对不同发育阶段的柞蚕,5龄起蚕的抗毒力比3龄起蚕增长47倍;5龄不同发育时期的柞蚕抗毒力的增长,与其体重增长有关。(6)不同外界环境,可导致柞蚕形成不同的生理状态,因此对核型多角体的抵抗力有明显差异;不同卵期经过的柞蚕抗毒力是:20天>30天>40天;不同饲料育的1、2龄柞蚕,不论是先食叶后添毒,或者是先添毒后食叶,其抗毒力均为适熟叶育>幼嫩叶育。     

柞蚕蛹的营养价值及综合利用研究进展

柞蚕蛹是一种优质、天然的蛋白源,具有极高的营养和药用价值,被广泛应用于食品、医药及农业等领域.本文综述了柞蚕蛹的营养价值,综述了柞蚕蛹深加工技术,包括蛹蛋白多肽、蚕蛹油脂、蛹虫草、免疫活性物质、生物防治方面应用,综述了柞蚕蛹生物学活性研究,展望了柞蚕蛹综合利用的发展前景,为其进一步开发应用提供参考.     

柞蚕蛹茶肠加工工艺的研究

柞蚕蛹内含有丰富的蛋白质、不饱合脂肪酸、矿物质、维生素等营养物质。因其滋味鲜美、低脂肪、高蛋白、无污染而深受人们的喜爱。柞蚕蛹的吃法很多,可以煮、炸、烧、烤、涮。本试验是用柞蚕鲜蛹蛋白替代茶肠中的部分精肉生产柞蚕蛹茶肠。经一系列加工,生产出的柞蚕蛹茶肠,口感好,具有独特的蛹香味,是老幼皆宜的食品。其加工工艺如下：     

柞蚕蛹血淋巴多酚氧化酶活性影响因素的研究

柞蚕血淋巴中多酚氧化酶活性的大小对柞蚕丝产量有一定影响,并对柞蚕蛹色起着非常重要的作用。本试验采用分光光度计法对柞蚕蛹血淋巴多酚氧化酶活性大小的影响因素进行了研究。结果表明:柞蚕蛹血淋巴多酚氧化酶的最适pH值为5.0;最适温度为25℃;少量的苯基硫脲对其的抑制作用较为明显;不同化合物对多酚氧化酶的活性影响不同,在相同浓度下,氯化钙和氯化镁对柞蚕蛹血淋巴多酚氧化酶的活性有促进作用,其他化合物(EDTA,维生素C和亚硫酸钠等)则表现为抑制作用。     

不同浓度Ca2+、Mg2+、Fe2+、Zn2+对紫花苜蓿生长的影响

以紫花苜蓿金皇后为材料,采用温室盆栽的方法对其在不同浓度Ca2+、Mg2+、Fe2+、Zn2+下的生长及结瘤情况进行探讨,结果表明:不同浓度不同离子对紫花苜蓿地上生物量及结瘤情况影响各有不同,其中Ca2+浓度为90mg/L时植株结瘤数目最丰,促进植株结瘤的作用最为明显;Mg2+浓度80mg/L时植株根冠比最大,为41.56％;Fe2+对植株结瘤数与蛋白质含量的影响作用最不明显,浓度为15mg/L时植株地上生物量干重为0.41g/株,增产作用优于Ca2+和Mg2+;Zn2+浓度为2.0mg/L时地上生物量干重(0.56g/株)和粗蛋白质含量(18.78％)均最高,对植株产量增加与蛋白质含量累积作用最为显著.     

添加柞蚕蛹"香肚"的加工工艺

柞蚕蛹内含有丰富的蛋白质、不饱和脂肪酸、矿物质、维生素等营养物质.近年来,因其滋味鲜美、低脂肪、高蛋白、无污染而越来越受到人们的喜爱.蚕蛹的吃法很多,可以煮、炸、烧、烤、涮.本试验是以柞蚕蛹蛋白替代肚制品中的部分精肉,经一系列加工而生产出的柞蚕蛹香肚,其口感好,风味独特,是老幼皆宜的绿色食品.现将加工工艺报道如下.     

重金属Cu2+、Cd2+和Pb2+对8种禾草种子萌发和幼苗生长的影响

研究了不同浓度Cu²⁺（200、300、400和500 mg·L^-1）、Cd²⁺（1、50、150和200 mg·L^-1）、Pb²⁺（500、1 000、1 500和2 000 mg·L^-1）对8种禾本科牧草种子萌发和幼苗生长的影响。结果表明：重金属离子胁迫对禾本科植物种子的发芽和幼苗生长的影响因植物种类及重金属离子不同而有差异。随着Cu²⁺,Cd²⁺和Pb²⁺浓度的增大,8种禾本科草种的相对发芽率、相对发芽指数和相对活力指数均呈不同程度的下降。供试不同浓度的Cu²⁺,Cd²⁺和Pb²⁺对8种植物幼苗生长均有抑制作用,高浓度胁迫下会出现“无根苗”。通过采用性状相对值对本试验进行隶属函数值综合评价,8种禾本科植物耐Cu²⁺的强弱顺序为：玉米 > 高丹草 > 燕麦 > 黑麦 > 无芒雀麦 > 黑麦草 > 垂穗披碱草 > 高羊茅,耐Cd²⁺的强弱顺序为：高丹草 > 黑麦 > 玉米 > 燕麦 > 无芒雀麦 > 垂穗披碱草 > 黑麦草 > 高羊茅,耐Pb²⁺的强弱顺序为：高丹草 > 燕麦 > 玉米 > 黑麦 > 无芒雀麦 > 垂穗披碱草 > 黑麦草 > 高羊茅。     

部分一化性柞蚕品种对微粒子病食下感染的抵抗性研究

<正>不同类型的柞蚕,对核型多角体病存在着不同的抵抗性[1],而对微粒子病的抵抗性如何,至今未见报导。近几年,柞蚕微粒子病的危害日趋严重,查明不同类型柞蚕对微粒子病的抗性差异,对养蚕防病和抗病育种等将会起到一定的指导作用。2002～2003年,我们用一化性柞蚕作材料,测定了部分蚕品种(生产种)和不同发育阶段的柞蚕对微粒子病的抵抗性。现将结果初报如下： 1 试验材料和方法 1．1 供试品种 三三、三九、一○一、河四一、早一、六号,均由本场柞蚕育种室品保组提供。 1．2 供试病原 取新鲜的柞蚕微粒子病蛹,加入适量灭菌水,通过研磨、过滤和离心提纯,将提纯的微孢子稀释、查数,配成一定浓度,保存于冰箱中备用。 1．3 测定方法 取不同类型的柞蚕作材料,在各龄起蚕时,将目的病液用微量进样器进行经口(蚁蚕     

Temperature and metal ions-dependent activity of the family I inorganic pyrophosphatase from the swine roundworm Ascaris suum

Temperature dependence, heat stability and metal ions-dependent activity were examined on the Family I inorganic pyrophosphatase (PPase) recently identified from Ascaris suum. Recombinant A. suum PPase (rAsPPase) showed an optimal activity at 55 degrees C. The rAsPPase was heat stable at 40 degrees C in the absence of added Mg(2+) and at 50 degrees C in its presence. The enzyme required divalent metal ions for its activity. The preferences for the metal ions (5 mM concentration) were in the order: Mg(2+)> Co(2+)> Cu(2+)> Fe(2+)> Zn(2+)> Mn(2+). On the contrary, enzyme activity was inhibited by Ca(2+). These findings suggest that catalytic features of AsPPase are consistent with the Family I PPases reported from a wide range of organisms.     

N素营养对NaCl胁迫下盐地碱蓬幼苗生长及渗透调节物质变化的影响

通过NaCl和N对盐地碱蓬幼苗生长及渗透调节物质变化的影响研究,结果表明,NaCl和N素对植物的生长的影响是明显的(P<0.05),低NaCl胁迫下N含量的增加对盐地碱蓬幼苗生长的影响不明显,而高NaCl处理下增加营养液中N的含量能明显地缓解盐胁迫,促进盐地碱蓬幼苗的生长.叶片脯氨酸含量随着NaCl浓度和养分N含量的增加而增加,而可溶性糖含量减少. NaCl浓度的增加使得植株总离子含量、Na 含量增加,K 、Ca2 、Mg2 含量减少.随N养分含量的增加,叶中Na 和K 含量增加, Mg2 含量基本上呈减少的趋势.叶片中的Ca2 含量和植株根茎部各离子含量随养分N含量增加的变化规律不明显.     

地衣芽孢杆菌CP-16脂类水解酶的研究

本试验旨在通过异源表达获得地衣芽孢杆菌(Bacillus licheniformis)CP-16的脂类水解酶,并探究其在羽毛降解过程中的作用。试验以地衣芽孢杆菌CP-16基因组DNA为模板,扩增脂类水解酶基因,转化入大肠杆菌中表达,获得重组酶L-4。研究重组酶L-4的适宜p H、p H稳定性、适宜温度、温度稳定性以及有机溶剂和金属离子对其相对活性的影响,同时探究其对角蛋白酶K水解天然羽毛角蛋白的作用。结果显示,获得的脂类水解酶基因大小为747 bp,编码248个氨基酸,在大肠杆菌中成功表达出重组酶L-4,其分子质量约为28.3 ku,酯酶活性为0.42 U/m L,适宜p H为6.5,适宜温度为50℃;在p H 6.5~9.5条件下处理30 min相对活性保持80%以上,在低于50℃温度条件下处理30 min相对活性保持70%以上。二价铁离子(Fe~(2+))、钠离子(Na~+)、锰离子(Mn~(2+))、钙离子(Ca~(2+))对重组酶L-4相对活性具有激发作用,钡离子(Ba~(2+))、锌离子(Zn~(2+))、铜离子(Cu~(2+))、镍离子(Ni~(2+))对重组酶L-4相对活性具有抑制作用。当有机溶剂浓度为30%时,重组酶L-4在二甲基亚砜(DMSO)和甲醇中保存97%和85%的相对活性,在丙酮、乙醇中保存45%以上的相对活性,在异丙醇中保存不到20%的相对活性,而在乙腈中相对活性基本完全丧失。用重组酶L-4预处理天然羽毛底物,可提高角蛋白酶K对底物的水解效率,促进率为4.32%。由此可见,脂类水解酶可降解羽毛表层脂质,可在促进角蛋白酶水解羽毛角蛋白中发挥作用。     

Improved pulmonary adaptation in newborn calves with postnatal acidosis.

The adaptation of newborn calves to extra-uterine life was evaluated by measuring arterial blood gases, acid-base values, blood ions and lung mechanical function parameters in normal and acidotic calves during the first 24 h. Twenty-seven Holstein Friesian newborn calves were divided into two groups according to their immediate post partum arterial blood pH values (Group A blood pH > or = 7.2: normal group; Group B blood pH between 7.2 and 7.0: acidotic group). Pulmonary function parameters were measured and arterial blood samples were analysed for blood gases, acid-base variables and ion content immediately post partum (within 2 min) and then 6 and 24 h after calving.Lung resistance and maximal difference between pressure maximum and pressure minimum (max delta Ppl) decreased, while dynamic lung compliance increased significantly in both groups. Immediately post partum the lung resistance and max delta Ppl were significantly higher in the acidotic group than in the normal group. The arterial blood pH progressively compensated with time in both groups during the first 24 h and there was no difference in arterial blood pH values between the two groups 6 h after birth. These results showed that the compensation of acidosis was associated with the improvement in lung mechanics and these changes occurred mainly during the first 6 h of life. Moderate to pronounced acidosis did not affect the pulmonary adaptation negatively, although some respiratory mechanics parameters (max delta Ppl), blood pH and Ca(2+)ion concentrations remained significantly different between the normal and acidotic groups at 24 h. This might be the result of overcompensation of acidosis and the interdependence between blood pH and Ca(2+)concentrations.     

Cu2+胁迫对多花黑麦草生长及饲草品质的影响

为探讨Cu2+污染对牧草生产的影响,盆栽研究了多花黑麦草的生长及品质对C2+胁迫的响应.结果表明多花黑麦草对Cu2+反应敏感,Cu2+处理浓度＞5mg/L时,株高、分蘖数及单株干物重急剧降低,生长显著受到抑制;Cu2+处理浓度＞30mg/L时,单株干物重降低至对照的39.0%.多花黑麦草植株体内的Cu2+积累量显著增加,但Cu2+在各器官中的分布存在明显的差异;Cu2+含量根＞茎＞叶,根中含量远远高于地上部.Cu2+处理浓度＞50mg/L时,茎叶粗蛋白(CP)含量随处理浓度的提高呈上升趋势,但Cu2+处理浓度＞50mg/L后,处理浓度提高导致饲草品质迅速下降.干物质量与CP含量的乘积,即CP养分产量在15mg/L Cu2+处理浓度时与对照无明显差异,到30mg/L Cu2+处理浓度时则急剧降为对照的42.2%.综合考虑Cu2+对多花黑麦草生长及品质的影响,土壤溶液中Cu2+＞30mg/L的污染土壤不宜进行多花黑麦草生产.     

Evaluation of a microwave method for dry matter determination in faecal samples from weaned pigs with or without clinical diarrhoea

Microwave drying as a procedure for determination of faecal dry matter in weaned pigs was evaluated and clinical relevant cut-off values between faecal consistency scores were determined. Repeatability and reproducibility were evaluated. Overall coefficient of variation was 0.03. The 95% confidence limits for any future faecal subsample examined by any operator in any replica were ± 0.85% faecal dry matter. Robustness in relation to weight of wet faeces was evaluated. The weight categories were 0.5, 1.0, 1.5, 2.0 and 3.0 g. Samples of 0.5 g gave significantly different mean faecal dry matter content compared to weighing of 1.0-3.0 g. Agreement with freeze-drying was evaluated. Lin's concordance correlation coefficient was 0.94. On average the faecal dry matter values was 1.7% (SD=1.99%) higher in freeze dried compared to micro waved samples. Non-parametric ROC analyses were used to determine optimal faecal dry matter cut-off values for clinical faecal consistency scores. The 4 consistency scores were score 1=firm and shaped, score 2=soft and shaped, score 3=loose and score 4=watery. The cut-off values were score 1: faecal dry matter content >19.5%, score 2: faecal dry matter content ≤ 19.5% and >18.0%, score 3: faecal dry matter content ≤ 18.0% and >11.3%, score 4: faecal dry matter content ≤ 11.3%. In conclusion, the microwave procedure has an acceptable repeatability/reproducibility and good agreement with freeze drying can be expected. A minimum of 1.0 g of wet faeces must be used for analyses. Faecal dry matter cut-off values between 4 different clinical consistency scores were determined.     

液相色谱-三重四极杆/线性离子阱复合质谱技术检测牛可食性组织中莫昔克丁的残留

建立了一种可准确定性定量检测4种牛可食性组织中莫昔克丁残留的液相色谱-三重四极杆/线性离子阱(LC-Qtrap)复合质谱分析技术。牛肌肉、肝脏、肾脏和脂肪样品经乙腈提取,高速离心去除蛋白质等杂质,C18柱净化。以0.1%甲酸水溶液和0.1%甲酸乙腈溶液为流动相进行洗脱,在BEH C18色谱柱上实现分离,在电喷雾正离子(ESI+)模式下,用多反应监测联合信息依赖性采集与增强子离子扫描(MRM-IDA-EPI)模式检测,基质匹配标准溶液外标法定量。结果表明:莫昔克丁在牛肌肉和牛肾脏1~100 ng/mL的基质匹配浓度范围内;在牛肝脏1~200 ng/mL的基质匹配浓度范围内和在牛脂肪1~1000 ng/mL的基质匹配浓度范围内均呈现良好的线性关系,相关系数(R 2)均大于0.990;在4种牛可食性组织中莫昔克丁的定量限均为2μg/kg。莫昔克丁在牛肌肉2~40μg/kg、牛肝脏2~200μg/kg、牛肾脏2~100μg/kg和牛脂肪2~1000μg/kg添加浓度水平上的回收率范围为65.6%~115%;批内与批间相对标准偏差均小于15%。该方法具有简便快速、灵敏度高、定性准确,重复性好等特点,可以满足上述组织中莫昔克丁残留检测的要求。     

Evaluation of farmers’ diagnostic performance for detection of diarrhoea in nursery pigs using digital pictures of faecal pools

Background

Overconsumption of antibiotics in the pig industry is of concern in relation to antimicrobial resistance. False positive disease diagnosis may result in the treatment of healthy animals. In Denmark, diarrhoea is the most common cause of antibiotic treatment in pigs. Farm personnel are not professional clinicians, which could result in inappropriate antibiotic treatments of diarrhoea.The primary objectives of this pilot study using digital pictures of faecal pools was to evaluate farmers’ diagnostic performance in the assessment of faecal consistency in nursery pigs and to investigate the effect of different co-variables, including practical experience. A secondary objective was to compare the diagnostic performance of farmers with that of veterinarians.At a pig congress, observers (farm personnel and veterinarians) working professionally with pigs participated in a faecal consistency test consisting of 16 pictures of faecal pools (eight diarrhoeic and eight non-diarrhoeic). The faecal pools had previously been collected and subjected to faecal dry matter determination. The true status of the faecal pools was determined by the faecal dry matter content (diarrhoea: faecal dry matter ≤ 18%). The true status was used to evaluate the farmers’ and veterinarians’ diagnostic performance.

Results

A total of 119 farmers and 18 veterinarians were included in the statistical analysis. For the farmers, the mean proportion of faecal pools assessed as diarrhoeic was 0.48, the mean proportion of correctly classified faecal pools was 0.84, the mean diagnostic sensitivity was 0.83 and the mean diagnostic specificity was 0.86. Farmers with less than four years of practical experience detected clinical diarrhoea more accurately than farmers with more than four years of practical experience (p < 0.05). No significantly differences between farmers and veterinarians was observed (p > 0.20).

Conclusions

The results, using digital pictures of faecal pools, suggest that farmers and veterinarians have similar diagnostic performance in relation to diarrhoea. False positive classification of non-diarrhoeic pigs appears to be a larger problem than false negative classification of diarrhoeic pigs under Danish conditions. If these results can be confirmed under practical conditions, training in, and validation of, clinical diagnoses may be an important factor in reducing antibiotic consumption in the pig industry.     

Clinical Applications of Quantitative Acid-Base Chemistry

Stewart used physicochemical principles of aqueous solutions to develop an understanding of variables that control hydrogen ion concentration (H+) in body fluids. He proposed that H+ concentration in body fluids was determined by PCO₂, strong ion difference (SID = sum of strong positive ion concentrations minus the sum of the strong anion concentrations) and the total concentration of nonvolatile weak acid (A_tot) under normal circumstances. Albumin is the major weak acid in plasma and represents the majority of A_tot. These 3 variables were defined as independent variables, which determined the values of all other relevant variables (dependent) in plasma, including H+. The major strong ions in plasma are sodium and chloride. The difference between Na+ and Cl^- may be used as an estimation of SID. A decrease in SID below normal results in acidosis (increase in H+) and an increase in SID above normal results in alkalosis (decrease in H+). Unidentified strong anions such as lactate will decrease the SID, if present. Equations developed by Fencl allow Stewart's work to be easily applied clinically for evaluating the metabolic (nonrespiratory) contribution to acid-base balance. This approach separates the net metabolic abnormality into components, and allows one to easily detect mixed metabolic acid-base abnormalities. The Fencl approach provides insight into the nature and severity of the disturbances that exist in the patient. Sodium, chloride, protein, and unidentified anion derangements may contribute to the observed metabolic acid-base imbalance.     

东祁连山高寒植被土壤解磷菌筛选及其解磷能力的初步研究

采用PKO固体培养基从东祁连山高寒草地中分离出具有解磷能力的菌株33株,对其进行继代培养,得到13株功能稳定的解磷菌,采用2种方法测定其解磷能力。13株解磷菌在PKO平板上8 d后,解磷圈D/d值为1.21~4.01;在液体摇瓶中,其解磷率在0.13~11.842 mg/g。其中,菌株2-2-1具有较高的解磷率,解磷效果在1%水平下显著高于其他菌株,具有较大的应用潜力。