Phenotypic and allelic variation for wort protein Z in Australian and Canadian barleys

Protein Z is a major component in beer foam. Two-dimensional electrophoresis was used to analyze wort proteins of two Australian (Buloke and Commander) and two Canadian (CDC Meredith and Bentley) varieties. The Canadian barley contained more abundant proteins from MW 40–45 kDa (pI 5 to 7). These proteins were identified as either protein Z4 or protein Z7 using liquid chromatography–mass spectrometry. Full-length gene of protein Z4 and Z7 were sequenced from Canadian and Australian barleys. Sequence differences were identified in the coding region and upstream regions of the two genes, resulting in protein sequence and expression variations. Molecular markers were designed according to the indels in the upstream regions of protein Z4 and Z7 genes. These markers were highly correlated to wort protein Z content in Canadian and Australian varieties. The Canadian barleys contained ‘high level’ genotypes for protein Z4 and Z7 while most Australian barleys had ‘low level’ genotypes for protein Z4, Z7 or both. The markers identified in this study provide a valuable tool for the selection of protein Z alleles in marker-assisted breeding. Total protein Z content was assessed using different steeping conditions, and increasing air-rest time increased protein Z content in 15 varieties.     

Effect of germination time and temperature on the functionality and protein solubility of sorghum flour

Sorghum was germinated for different time (12, 24, 36 and 48 h) at different temperatures (25, 30 and 35 °C) and the changes in their nutritional and functional properties of germinated sorghum flour were assessed and compared with native sorghum flour. Germination inversely affects the crude protein, fat, fibre and ash content. A decrease in water absorption and swelling power and increase in oil absorption capacity was observed due to enzymatic starch modification as the germination duration progressed. Germination of sorghum increased the gel consistency while paste clarity was decreased as compared to native flour. Proteins were modified by action of enzymes during higher germination time and temperature conditions, which results in significantly higher protein solubility of germinated sorghum flour, which also result in enhancing the foaming and emulsifying properties of the flour. Lowest % synersis value and least gelation concentrations were observed in native sorghum has, which increased during germination and were highest in sorghum germinated for 48 h at 35 °C. Germination in overall can be used as low cost natural bio-processing technique for the preparation of modified flour with enhanced function properties without chemical modification or genetic engineering.     

An improved method for extraction of sorghum polymeric protein complexes

A method for fractionating sorghum proteins using extraction solvents and techniques designed to obtain polymeric protein structures (especially disulfide linked) was developed. Extraction and separation conditions were optimized in terms of completeness of protein extraction, sample stability, and analytical resolution. After pre-extraction of albumins and globulins, a 3-step sequential procedure involving no reducing agents was applied to ground whole sorghum flour. The three fractions obtained represented proportionally different protein polymer contents and molecular weight distribution as evidenced by comparative size exclusion chromatography. Protein composition also varied among the extracts with differences in kafirin composition and non-kafirin proteins detected in the fractions by RP-HPLC and SDS-PAGE analysis. The ability to quantify and further characterize sorghum polymeric protein complexes will be useful for additional studies linking protein structures with functionality and digestibility and variations for these properties within diverse sorghum germplasm.     

抗凋亡融合蛋白PTD-Bcl-xL原核表达载体的构建及表达纯化

人工抗凋亡蛋白PTD-Bcl-x L能保护多种因素引起的细胞异常凋亡,为了获得高纯度Bcl-x L与PTD(Protein transduction domains)的融合蛋白,首先采用TRIzol法提取SD大鼠肝脏总RNA,将RNA反转录为c DNA,设计引物以c DNA为模板,PCR扩增Bcl-x L基因,构建p UM19-T-Bcl-x L质粒,并对质粒双酶切鉴定和测序鉴定;其次设计包含PTD序列的Bcl-x L引物,以测序正确的p UM19-T-Bcl-x L质粒为模板,PCR扩增PTD-Bcl-x L序列,将扩增序列克隆入p ET28a载体,构建PTD-Bcl-x L蛋白的原核表达质粒p ET28a-PTD-Bcl-x L,并对p ET28a-PTD-Bcl-x L载体双酶切鉴定和测序鉴定;将p ET28a-PTD-Bcl-x L重组质粒转化大肠杆菌BL21(DE3),用IPTG诱导表达,并对IPTG诱导融合蛋白表达的浓度和诱导时间进行了优化;SDS-PAGE分析表达蛋白的可溶性情况,在变性条件下用Ni-NTA琼脂纯化融合蛋白;最后用SDS-PAGE、Western Blot及质谱对融合蛋白进行鉴定。结果表明:双酶切p UM19-T-Bcl-x L质粒出现约774 bp大小条带,p UM19-T-Bcl-x L质粒测序结果与NCBI数据库比对序列一致,表明成功构建p UM19-T-Bcl-x L质粒;双酶切p ET28a-PTD-Bcl-x L质粒出现约744 bp大小条带,p ET28a-PTD-Bcl-x L质粒测序结果与预期序列一致,表明成功构建了p ET28a-PTD-Bcl-x L原核表达载体;在IPTG诱导下p ET28a-Bcl-x L重组质粒在大肠杆菌BL21(DE3)中表达出36 k Da大小蛋白,最优IPTG诱导浓度为0.1 mmol/L,最佳IPTG诱导时间为6 h;SDS-PAGE电泳显示融合蛋白主要出现在菌液超声后的沉淀里,以包涵体形式表达,经Ni-NTA琼脂纯化获得了高纯度的融合蛋白;Western Blot和质谱鉴定证明IPTG诱导表达蛋白和纯化的融合蛋白为PTD-Bcl-x L蛋白。纯化得到了PTD-Bc L-x L融合蛋白,推进了PTD-Bcl-x L蛋白在猪、牛等家畜精液冷冻保存的应用进程。     

用酶法制备藏羊血食用蛋白粉的研究

采用四因素三水平正交试验,对藏羊血制备食用蛋白的工艺进行了分析。经分析食用蛋白粉水解最佳条件为：酶解用酶量4.5g,酶解温度为46℃,酶解时间为7.5h,酶解的pH值为10.5,活性炭作用于水解液时由室温升至80℃所需最佳时间为6～8min。     

Protein and energy concentrations of meat meal and meat and bone meal fed to pigs based on in vitro assays

The objective of this study was to develop equations for estimating ileal digestible crude protein (CP) and metabolizable energy (ME) contents of meat meal (MM) and meat and bone meal (MBM) as feed ingredients for pigs based on in vitro assays. Test ingredients were 4 sources of MM and 3 sources of MBM. Ash and CP contents of the ingredients ranged from 3.8% to 33.1% and 46.8% to 82.9% (as-is basis), respectively. In vitro ileal disappearance (IVID) of CP was determined and ileal digestible CP content was calculated by multiplying CP content by IVID of CP. In vitro total tract disappearance (IVTTD) of dry matter (DM) was determined and ME was calculated using gross energy, CP contents, and IVTTD of DM. The IVID of CP and IVTTD of DM ranged from 77.2% to 88.7% and from 82.7% to 92.4%, respectively. Calculated ileal digestible CP and ME contents ranged from 37.8% to 73.5% DM and 2,405 to 3,905 kcal/kg DM, respectively. Ash contents were negatively correlated (P < 0.001) with CP (r = −0.99), in vitro ileal digestible CP (r = −0.97), gross energy (r = −1.00), in vitro digestible energy (r = −0.97), and adjusted ME (r = −0.97). The most fitting equations for ileal digestible CP and adjusted ME were: ileal digestible CP (% DM) = 11.91 − 0.90 × Ash (% DM) + 0.74 × IVID of CP (%) (R² = 0.99) and adjusted ME (kcal/kg DM) = 130.85 − 50.90 × ash (% DM) + 47.06 × IVTTD of DM (%) (R² = 0.99). To validate the accuracy of the prediction equations for ME, mean bias and linear bias were determined using a regression analysis. Calculated ME values of MM and MBM were in a good agreement with data obtained from animal experiments based on a statistically insignificant bias in the models. In conclusion, ME concentrations of MM and MBM as swine feed ingredients can be calculated using ash concentration and in vitro disappearance of dry matter.     

Comparative pharmacokinetics of marbofloxacin in healthy and Mannheimia haemolytica infected calves

The pharmacokinetics of marbofloxacin were investigated in healthy (n=8) and Mannheimia haemolytica naturally infected (n=8) Simmental ruminant calves following intravenous (i.v.) and intramuscular (i.m.) administration of 2 mg kg(-1) body weight. The concentration of marbofloxacin in plasma was measured using high performance liquid chromatography with ultraviolet detection. Following i.v. administration of the drug, the elimination half-life (t(1/2 beta)) and mean residence time (MRT) were significantly longer in diseased calves (8.2h; 11.13 h) than in healthy ones (4.6 h; 6.1 h), respectively. The value of total body clearance (CL(B)) was larger in healthy calves (3 ml min(-1) kg(-1)) than in diseased ones (1.3 ml min(-1) kg(-1)). After single intramuscular (i.m.) administration of the drug, the elimination half-life, mean residence time (MRT) and maximum plasma concentration (C(max)) were higher in diseased calves (8.0, 12 h, 2.32 microg ml(-1)) than in healthy ones (4.7, 7.4 h, 1.4 microg ml(-1)), respectively. The plasma concentrations and AUC following administration of the drug by both routes were significantly higher in diseased calves than in healthy ones. Protein binding of Marbofloxacin was not significantly different in healthy and diseased calves. The mean value for MIC of marbofloxacin for M. haemolytica was 0.1+/-0.06 microg ml(-1). The C(max)/MIC and AUC(24)/MIC ratios were significantly higher in diseased calves (13.0-64.4 and 125-618 h) than in healthy calves (8-38.33 and 66.34-328 h). The obtained results for surrogate markers of antimicrobial activity (C(max)/MIC, AUC/MIC and T > or = MIC) indicate the excellent pharmacodynamic characteristics of the drug in diseased calves with M. haemolytica, which can be expected to optimize the clinical efficacy and minimize the development of resistance.     

小麦杂种后代籽粒蛋白质含量的配合力研究

选用６个蛋白质含是高低不同的亲本，采用Ｇｒｉｆｆｉｎｇ双列杂交方法二，对亲本在Ｆ１－Ｆ３的蛋白质含量配合力作了研究。结果表明，蛋白质含量的ＧＣＡ与ＳＣＡ方差均极显著，表明在要试验中基因加性效应和非加性效应均起重要作用。但由于三个世代ｇｃａＭｓ／ｓｃａＭｓ值均极显著，且随着世代的推进这一比值又逐渐增加，因此蛋白质含量在杂种后代的表现主要还是由基因加性效应决定，随着世代的推进，基因加性效应越显重要，在     

Antibody response to Raboral VR-G® oral rabies vaccine in captive and free-ranging black-backed jackals (Canis mesomelas)

Rabies is a zoonotic disease that remains endemic in large parts of southern Africa because of its persistence in wildlife and domestic dog vectors. The black-backed jackals (Canis mesomelas) is primarily the wildlife vector responsible for rabies outbreaks in northern parts of South Africa. Two trials were carried out to investigate antibody responses to the oral rabies vaccine Raboral V-RG® in black-backed jackals under captive and free-ranging conditions. In captive jackals 10/12 (83%; 95% confidence interval [CI]: 52% – 98%), seroconverted after single oral vaccination. Nine captive jackals had protective antibody titres (> 0.5 IU/mL) at 4 weeks (median: 2.1 IU/mL; inter quartile range [IQR]: 0.6–5.7) and 10 jackals had at 12 weeks (median: 3.5 IU/mL; IQR: 1.5–8.3) and three maintained antibody titres for up to 48 weeks (median: 3.4 IU/mL; IQR: 2.0–6.3). Four sites were baited with Raboral V-RG® vaccine for wild jackals, using fishmeal polymer and chicken heads. Baits were distributed by hand or from vehicle at three sites in north-eastern South Africa, with an average baiting density of 4.4 baits/km² and at one site in central South Africa, at 0.12 baits/km². This resulted in protective antibody titres in 3/11 jackals (27%; 95% Cl: 6–61) trapped between 3 and 12 months after baiting in north-eastern South Africa, compared with 4/7 jackals (57%; 95% Cl: 18–90) trapped after 3–18 months in central South Africa. This study shows the potential utility of oral rabies vaccination for the control of wildlife-associated rabies in north-eastern and central South Africa, but extensive studies with wider distribution of bait are needed to assess its potential impact on rabies control in wild jackals.