电子鼻和电子舌单独与联合检测掺大豆蛋白或淀粉的鸡肉糜

为实现掺杂掺假鸡肉的快速、客观评价，该研究利用电子鼻和电子舌联合检测技术对掺杂鸡肉糜进行快速检测，通过对采集的数据进行主成分分析和偏最小二乘法分析，所得结果表明，采用主成分分析，电子鼻和电子舌联合检测掺大豆蛋白鸡肉糜和掺淀粉鸡肉糜的主成分总贡献率分别为99.8%和99.1%；采用偏最小二乘法分析，电子鼻和电子舌联合检测鸡肉糜中掺杂大豆蛋白含量的预测值与真实值之间的决定系数为0.992，均方根误差为2.8%；联合检测鸡肉糜中掺杂淀粉含量的预测值与真实值之间的决定系数为0.996，均方根误差为2.4%。表明电子鼻和电子舌联合检测对鸡肉糜的掺杂情况具有良好的区分和预测能力，并且是一种有效、高精度的肉类掺假检测方法。     

Bt抗虫棉新品系毒蛋白表达差异分析

【目的】本研究旨在探讨部分抗虫棉不抗虫的原因。【方法】通过室内生物测定、卡那霉素、Bt-Cry1Ab/Ac试纸条、PCR分析、Southern blot检测和Bt蛋白含量检测等方法分析了23个抗虫棉新品系的抗虫性。【结果】生测结果表明供试品系间幼虫死亡率差异显著。卡那霉素、Bt-Cry1Ab/Ac试纸条、PCR分析和Southern blot检测结果表明,Bt基因在供试材料中整合并稳定遗传且为单拷贝。RT-PCR结果表明,各试验品系Bt基因相对表达量差异较大,花期叶片Bt基因的相对表达量最高;Bt蛋白检测结果发现抗虫棉Bt蛋白表达量在生育前期大于后期,营养器官中的大于生殖器官,不同年份Bt蛋白含量差异显著。幼虫死亡率与Bt基因相对表达量的相关系数仅为0.1745,与Bt蛋白的相关系数为0.7130,表明Bt蛋白含量越高,抗虫性越好;各组织器官的Bt蛋白与Bt基因的相对表达量相关系数为-0.3659~0.2542,二者表达趋势不一致,表明遗传背景对抗虫棉Bt蛋白的表达具有一定影响。且Bt基因可能在转录后受到调控导致Bt蛋白含量发生变化,从而影响了抗虫棉的抗虫性。【结论】这些结果有望为抗虫棉育种提供参考。     

饲料蛋白质和脂肪水平对亚东鲑亲鱼生长性能、消化酶活性和血清指标的影响

本试验旨在研究饲料蛋白质和脂肪水平对亚东鲑亲鱼生长性能、性腺指数、消化酶活性和血清指标的影响。试验采用5×2两因素随机设计,设5个蛋白质水平(36%、39%、42%、45%、48%)和2个脂肪水平(9%、18%),共配制10种实用试验饲料。每种试验饲料投喂3个重复,每个重复19尾鱼。初重为(462.53±45.40)g亚东鲑在室内水族箱中流水饲养,饲养周期为77 d。结果表明:同一脂肪水平下,随饲料蛋白质水平的升高,增重率有所升高,而饲料系数逐渐降低,至蛋白质水平为48%时又有所升高。饲料蛋白质和脂肪水平对雌性亲鱼性腺指数无显著影响(P0.05),雄性亲鱼性腺指数随蛋白质水平的升高而降低,但至蛋白质水平为48%时又有所回升。同一脂肪水平下,随着饲料蛋白质水平的升高,胃、肠道和幽门盲囊的脂肪酶活性逐渐增加,蛋白质水平为48%时显著高于蛋白质水平为36%时(P0.05);而胃、肠道和幽门盲囊的蛋白酶活性虽有所增加,但各组间无显著差异(P0.05)。同一蛋白质水平下,18%脂肪水平组胃、肠道和幽门盲囊的蛋白酶和脂肪酶活性均略高于9%脂肪水平组。同一脂肪水平下,血清氨含量随饲料蛋白质水平的升高而逐渐增加。饲料蛋白质和脂肪水平对血清超氧化物歧化酶、溶菌酶活性以及葡萄糖、甘油三酯、总蛋白、白蛋白和丙二醛含量均未产生显著影响(P0.05)。由此得出,亚东鲑亲鱼对饲料蛋白质(36%~48%)和脂肪(9%~18%)水平有一定的适应性反应;以增重率为指标进行折线模型分析,得出饲料脂肪水平为9%和18%时,亚东鲑亲鱼对蛋白质需要量分别为43.11%和45.69%。     

Reducing crude protein variability and maximizing savings when formulating corn-soybean meal-based feeds

Crude protein in corn and soybean meal have been documented to vary, and such inherent variability can result in under- or over-feeding of CP when feeds are formulated, leading to reduced bird growth, added input costs, and increased environmental pollution. The purpose of this study was to compare 2 grain-handling techniques and 2 feed formulation methods (linear vs. stochastic programming) to reduce CP variability in finished feeds and determine resulting costs or savings. The 2 grain-handling techniques were placing all the random batches of each delivered ingredient in to (1) a single bin (1-bin method) or (2) segregating above- and below-average samples into 2 bins (2-bin method). A fast way of estimating the composition of the ingredients is now available (near-infrared reflectance spectroscopy). Microsoft Excel workbooks were constructed to solve broiler starter feed formulation problems. Formulating feeds by linear and stochastic models based on the 2-bin method reduced CP variability by at least 50% compared with the 1-bin method. Formula cost was reduced by ˜20 cents per ton (averages of August 2012 United States ingredient prices) when the 2-bin method was used with the linear model. Formulating feed with a margin of safety increased formula cost by $3.40 per ton. Stochastic feed formulation increased formula cost to meet the specified CP level in feed at any probability of success, and formula cost was reduced substantially with the 2-bin method (up to $6.47 per ton). The magnitude of savings and reduced feed variability suggested that, regardless of the costs associated with building extra bins, the 2-bin method can be economically efficient in the long run. Therefore, it could be possible to split the batches of feed ingredients at a feed mill into above- or below-average bins before feed formulation to reduce CP variability and to maximize savings.     

基于S基因序列分析新型猪流行性腹泻病毒的遗传演化

最近，笔者实验室在青藏高原地区发现两种新亚型藏猪源猪流行性腹泻病毒（PEDV），为进一步调查新型PEDV是否在四川腹泻猪群中存在或流行，对实验室2018-2019年保存的116份猪腹泻粪便或肠组织样本进行PEDV的检测及其纤突蛋白基因（spike）分子特征研究。结果表明：腹泻样本的PEDV检出率为42.2%（49/116，95% CI=33.1%~51.8%），并获得了13条完整的S基因序列，全长为4 149~4 170 bp，序列相似性为94.2%~99.9%，其中SWUN-H3-CH-SCYA-2019的S基因与藏猪源新G1亚群PEDV的序列相似性高达97.0%~98.6%。遗传演化研究结果表明13株PEDV S基因划分为G1和G2大群，其中SWUN-H3-CH-SCYA-2019位于藏猪源新G1亚群；SWUN-19-CH-SCZY-2018、SWUN-4-CH-SCXC-2018、SWUN-1-CH-SCNJ-2019和SWUN-3CH-CH-SCZG-2019位于G2亚群中一个独立的分支，且与藏猪源新G2亚群毒株有着较近的亲缘关系。为了进一步研究13株PEDV的演化过程，以贝叶斯进化分析软件包（BEAST）进行分歧时间估算，结果表明SWUN-H3-CH-SCYA-2019的分歧时间约为2012.3年，早于藏猪源新G1亚群其余毒株的最早分歧时间（2015.7年）；SWUN-4-CH-SCXC-2018、SWUN-19-CH-SCZY-2018和SWUN-3CH-CH-SCZG-2019的分歧时间约为2014.2年，早于G2亚群的藏猪源毒株2014.7年，所有藏猪源PEDV的分歧时间均晚于四川毒株。本研究在四川地区首次发现了藏猪源PEDV，并且从毒株的分歧时间推断青藏高原的藏猪源PEDV来源于四川，为新型PEDV分子遗传进化的监测提供了依据。     

Application of urease inhibitor improves protein composition and bread‐baking quality of urea fertilized winter wheat

Background: Nitrogen losses is an economic problem for wheat production and a high risk to the environment. Therefore, improved N fertilizer management is a key to increasing the N efficiency and minimizing N losses. To increase N efficiency, enhanced fertilizers such as urea combined with urease inhibitor can be used. Aims: The aim of present study was to evaluate the effects of different N forms on grain storage protein subunits in winter wheat and to examine whether the observed changes correlate with parameters of baking quality. Methods: The investigation was performed over two consecutive years at two locations in Germany. Protein subunits were analyzed by SDS‐PAGE. Results: Protein concentrations were similarly increased after fertilization with ammonium nitrate and urea + urease inhibitor. Analysis of the individual storage protein fractions indicated that both fertilizers specifically enhanced ω‐gliadins and HMW glutenins, but the effect was more pronounced in the ammonium nitrate treatment. Application of urea + urease inhibitor had greater influence on the protein composition and resulted in higher specific baking volume as well as the best fresh keeping ability, in comparison with urea treatment. Conclusion: Considering that the urea + urease inhibitor treatment resulted in almost comparable improvements of NUE and baking quality, with the additional benefit of reduced N losses in combination with easy handling, urea + urease inhibitor can be recommended as a viable alternative to both urea alone and ammonium nitrate treatments. This opens up an opportunity for the reduction of N loss in wheat production when use of urea is preferred.     

Effects of ZIP14 on biological behaviors of hepatocellular carcinoma cell line BEL-7404

AIM: To study the expression of zinc transporter ZRT/IRT-like protein 14 (ZIP14) in the hepatocellular carcinoma (HCC) tissues, and to investigate the effects of ZIP14 over-expression on the biological behaviors of HCC cells. METHODS: The expression of ZIP14 at mRNA and protein levels in the HCC tissues and adjacent non-tumor tissues were detected by real-time PCR and immunohistochemical staining, respectively. The lentivirus expression system containing GV365-ZIP14 was constructed, and was used to infect the HCC cell line BEL-7404, which had relatively poor expression of ZIP14. The expression of ZIP14 at mRNA and protein levels in the transfected cells were detected by real-time PCR and Western blot, respectively. Under the conditions of zinc sulfate stimulation at different concentrations, the cell viability, the cell cycle, and the cell migration and invasion abilities were detected by MTT assay, DNA ploid detection, and Transwell assay, respectively. RESULTS: The mRNA expression level and the strong-positive rate of protein expression of ZIP14 in the HCC tissues were significantly lower than those in the adjacent non-tumor liver tissues (P<0.01). The expression of ZIP14 at mRNA and protein levels in the BEL7404 cells was significantly enhanced by infection of GV365-ZIP14 expression lentivirus. Compared with negative control group (transfected with negative control lentivirus), the cell viability, migration and invasion in ZIP14 over-expression group (transfected with GV365-ZIP14 expression lentivirus) were significantly reduced, and the percentage of the cells in G₂/M phase was significantly increased, all of which were more obvious with the elevation of zinc concentration in the culture medium. CONCLUSION: ZIP14 is low expressed in the HCC tissues. The ZIP14 over-expression has inhibitory effects on the viability, migration and invasion of HCC cells, and blocks the cell cycle in G₂/M phase, which might be closely related to the elevation of zinc concentration in cytoplasma of HCC cells due to enchanced zinc transport by ZIP14.     

马链球菌兽疫亚种烯醇化酶对小鼠肺泡巨噬细胞吞噬功能的影响

旨在研究马链球菌兽疫亚种（Streptococcus equi ssp.zooepidemicus，SEZ）烯醇化酶（enolase，Eno）对小鼠肺泡巨噬细胞（RAW264.7）吞噬能力的影响。通过构建原核表达质粒获得重组烯醇化酶（rEno），采用台盼蓝活细胞计数法，判定在不同处理浓度和时间下rEno蛋白对RAW264.7细胞的细胞毒性。将rEno蛋白与RAW264.7细胞共孵育后，用SEZ作用于细胞并检测细胞吞菌数量，判断RAW264.7细胞对SEZ的吞噬活性。进一步通过活细胞稳定同位素标记技术（SILAC）和蛋白质谱分析技术（LC-MS/MS），筛选到RAW264.7细胞中可能与SEZ Eno存在相互作用的候选蛋白。结果发现，10 μg·mL^-1 rEno蛋白处理对RAW264.7细胞有明显的细胞毒性，且10 μg·mL^-1 rEno蛋白处理RAW264.7细胞2和4 h可显著抑制其对SEZ的吞噬作用（P<0.01、P<0.05）。初步筛选到RAW264.7细胞中动力蛋白激活蛋白亚单位蛋白（dynactin subunit protein 2，Dctn）、整合素α-M蛋白（integrin alpha-M）等17种可能与Eno发生互作的蛋白。本研究获得了rEno重组表达蛋白，发现rEno可减少RAW264.7细胞对SEZ的吞噬，互作蛋白的初步筛选也为进一步揭示Eno在SEZ抗吞噬中的作用机制奠定了基础。     

Outbreak of anaplasmosis associated with novel genetic variants of Anaplasma marginale in a dairy cattle

During early lactation, dairy cows may present a transient immunosuppressive state and develop anaplasmosis caused by Anaplasma marginale. In this study, clinical anaplasmosis in dairy cattle in the Thrace region of Turkey was investigated with respect to within-herd prevalence, vertical transmission, and genetic diversity. In March and September 2015, thirty lactating cows showed primary clinical signs of anaplasmosis, including fever, anaemia, decreased milk yield, anorexia, and laboured breathing. Symptoms disappeared in most cows after administration of long-acting oxytetracycline, but nine of them (30%) died. Following diagnosis based on clinical signs, microscopy and molecular findings, blood samples were collected from apparently healthy lactating cows (n = 184), pregnant heifers (n = 39) and newborn calves (n = 24). DNA was extracted from each sample and analyzed for the presence of major surface proteins (MSPs) of A. marginale, followed by sequencing to assess diversity of isolates. Microscopic examination of erythrocytes revealed A. marginale inclusion bodies in symptomatic cows. Examination of thin blood smears showed 3.8% of the lactating, clinically asymptomatic, cows to be infected with A. marginale, while nPCR detected 31.0% positive. A. marginale infection was not detected in pregnant heifers by either method. Congenital infection was found in one calf by nPCR. This is the first report of transplacental transmission of A. marginale in Turkey. The MSP4 sequence analyses showed high genetic diversity among the isolates, presenting 97.6-99.6% homology at the amino acid level. The sequences of MSP1a amplicons revealed genetic diversity providing three new tandem repeats.