双季稻最佳磷肥和钾肥用量与密度组合研究

【目的】为明确磷肥、钾肥用量和移栽密度对双季稻的施用效果,在田间试验条件下研究了不同磷肥用量、钾肥用量和移栽密度组合对江西双季稻产量、产量构成要素及磷肥和钾肥利用率的影响。【方法】本研究采用裂区试验设计研究了不同施磷量和移栽密度、不同施钾量和移栽密度对双季稻产量、磷肥和钾肥利用率的影响。磷肥用量和移栽密度试验中,设4个施磷水平(P2O5 0、60、90、120 kg/hm2,以P0、P60、P90和P120表示)和4种移栽密度(21×104、27×104、33×104、39×104 穴/hm2,以D21、D27、D33和D39表示)组合。钾肥用量和移栽密度试验中,设4个施钾水平(K2O 0、90、120、150 kg/hm2,以K0、K90、K120和K150表示),密度设置同磷肥试验。在水稻成熟期对产量以及产量构成要素进行测定,并分析其磷素和钾素的吸收量和利用率等指标。【结果】磷肥与密度试验中,同一施磷水平下,早稻产量和地上部磷素吸收量随着移栽密度的增加而增加,当施磷量超过60 kg/hm2时,产量和磷素吸收量不再随密度增加而显著增加,磷素吸收利用率(REP)、磷素农学效率(AEP)和磷素偏生产力(PFPP)逐步降低,以P60D39处理组合的产量和磷素吸收利用率最高,分别为5303.9 kg/hm2和24.4%,AEP为29.4 kg/kg; 晚稻则以施磷量在60 kg/hm2和33×104 穴/hm2密度组合的产量和磷素吸收利用率最高,分别为7246.9 kg/hm2和42.4%,AEP为36.2 kg/kg。钾肥与密度试验中,早稻的钾素吸收量随着施钾量的增加而增加,施钾量在120 kg/hm2和39×104 穴/hm2密度组合的处理产量和钾素吸收利用率(REK)最高,分别为6376.3 kg/hm2和67.2%,此时钾素农学效率(AEK)为15.6 kg/kg; 晚稻则以施钾量在90 kg/hm2和33×104 穴/hm2密度组合的处理产量和REK最佳,分别为7025.6 kg/hm2和74.0%,AEK为21.7 kg/kg。【结论】合理的磷肥、钾肥用量和移栽密度可以显著增加水稻单位面积有效穗数和养分累积量,进而增加水稻产量和肥料利用率,但过高的磷肥和钾肥施用会抑制产量的进一步增加。建议本研究区域的早稻采用施磷量在60 kg/hm2、施钾量120 kg/hm2和39×104穴/hm2的密度组合,而晚稻采用施磷量60 kg/hm2、施钾量90 kg/hm2和33×104 穴/hm2的密度组合。     

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。     

复杂疾病的遗传咨询技术

复杂疾病大面积爆发使人们的生活健康面临着重大挑战。在这一环境下,遗传咨询在人们的生活中占据了越来越重要的地位。依据基因与环境互作共同影响个体健康的原理,咨询者在遗传咨询师的指导帮助下合理解决各种遗传学问题可以有效降低复杂疾病的发病率。目前国内的遗传咨询发展尚处于萌芽期,随着遗传咨询在健康领域的地位上升,该技术在未来将会进一步推广应用。     

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。     

Critical comparison of the vanadomolybdate and the molybdenum blue methods for the analysis of phosphate in plant sap

Abstract

The performance of the vanodomolybdate and molybdenum blue methods for the routine determination of phosphate in plant sap were compared in order to assess their suitability for the detection of P deficiency at an early stage of plant growth. The molybdenum blue method was ca 10 times more sensitive than the vanadomolybdate one and required less sap to give a successful test. Recoveries of phosphate added to both sap and solution samples were close to 100% with both methods. The methods were in good agreement over a wide range of phosphate concentrations using sap samples from all leaves during the early stages of P deficiency, but increases in sap background colour caused the vanadomolybdate method to overestimate phosphate concentrations as the deficiency became more acute. Measurements on solutions of similar absorbance showed that the vanadomolybdate method was not significantly more variable than the molybdenum blue method but tended to overestimate phosphate concentration when very small volumes of sap were available. It was concluded that the molybdenum blue method was the most reliable for routine measurements of phosphate in plant sap.     

Identification and characterization of the enzymes responsible for the metabolism of the non‐steroidal anti‐inflammatory drugs,flunixin meglumine and phenylbutazone,in horses

The in vivo metabolism and pharmacokinetics of flunixin meglumine and phenylbutazone have been extensively characterized; however, there are no published reports describing the in vitro metabolism, specifically the enzymes responsible for the biotransformation of these compounds in horses. Due to their widespread use and, therefore, increased potential for drug–drug interactions and widespread differences in drug disposition, this study aims to build on the limited current knowledge regarding P450‐mediated metabolism in horses. Drugs were incubated with equine liver microsomes and a panel of recombinant equine P450s. Incubation of phenylbutazone in microsomes generated oxyphenbutazone and gamma‐hydroxy phenylbutazone. Microsomal incubations with flunixin meglumine generated 5‐OH flunixin, with a kinetic profile suggestive of substrate inhibition. In recombinant P450 assays, equine CYP3A97 was the only enzyme capable of generating oxyphenbutazone while several members of the equine CYP3A family and CYP1A1 were capable of catalyzing the biotransformation of flunixin to 5‐OH flunixin. Flunixin meglumine metabolism by CYP1A1 and CYP3A93 showed a profile characteristic of biphasic kinetics, suggesting two substrate binding sites. The current study identifies specific enzymes responsible for the metabolism of two NSAIDs in horses and provides the basis for future study of drug–drug interactions and identification of reasons for varying pharmacokinetics between horses.     

“1237”P氏生物安全立体非洲猪瘟防控体系

在目前非洲猪瘟疫苗毒株、变异毒株、野毒株并存背景下,只做好外部生物安全已不足以达到防控非洲猪瘟的目的。因疫苗毒株隐蔽性强、检测难度大,不知道通过引种是否会将疫苗毒株带到猪场。因此,未来猪场生物安全不能只关注场外生物安全,更重要的是要运用“以猪为本”的“1237”P氏生物安全立体非洲猪瘟防控体系,即在做好“控五流”等外部生物安全的同时,更要关注饮水安全、注重猪的自身健康度、提高猪的非特异性免疫力,才能实现“与非共存,与蓝共舞”。所谓“立体防控”是指全方位的防控,而不是仅仅针对猪场大门的传统生物安全防控。     

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.