Rapidly quantitative detection of Nosema ceranae in honeybees using ultra-rapid real-time quantitative PCR

BackgroundThe microsporidian parasite Nosema ceranae is a global problem in honeybee populations and is known to cause winter mortality. A sensitive and rapid tool for stable quantitative detection is necessary to establish further research related to the diagnosis, prevention, and treatment of this pathogen.ObjectivesThe present study aimed to develop a quantitative method that incorporates ultra-rapid real-time quantitative polymerase chain reaction (UR-qPCR) for the rapid enumeration of N. ceranae in infected bees.MethodsA procedure for UR-qPCR detection of N. ceranae was developed, and the advantages of molecular detection were evaluated in comparison with microscopic enumeration.ResultsUR-qPCR was more sensitive than microscopic enumeration for detecting two copies of N. ceranae DNA and 24 spores per bee. Meanwhile, the limit of detection by microscopy was 2.40 × 10⁴ spores/bee, and the stable detection level was ≥ 2.40 × 10⁵ spores/bee. The results of N. ceranae calculations from the infected honeybees and purified spores by UR-qPCR showed that the DNA copy number was approximately 8-fold higher than the spore count. Additionally, honeybees infected with N. ceranae with 2.74 × 10⁴ copies of N. ceranae DNA were incapable of detection by microscopy. The results of quantitative analysis using UR-qPCR were accomplished within 20 min.ConclusionsUR-qPCR is expected to be the most rapid molecular method for Nosema detection and has been developed for diagnosing nosemosis at low levels of infection.     

魔芋试管微球茎成熟度对其储藏及萌发率的影响

种子在母体植株上的干燥过程是种子完成从发育到萌发的一个重要转换点。魔芋组织培养中诱导试管微球茎过程中，在色泽上有1个明显的变化过程。本实验将这个过程分为3个阶段，对不同采收时期的微球茎进行储藏及萌发研究，结果表明，不同采收期的微球茎，储藏后的萌发率具有较大的差异，而且这种差异与魔芋种具有较大的相关性，这种差异与其保水性能有关。同时对试管微球茎收获度对储藏后萌发率影响的机制进行了初步的探讨。     

转基因抗虫棉中棉所76高效制种技术规程

总结了优质抗虫棉中棉所76高效制种技术,包括：制种田的选择、前期准备、亲本种植与管理、制种程序及注意事项等关键技术。     

Apparent ileal digestibility of nutrients and amino acids in soybean meal,fish meal,spray‐dried plasma protein and fermented soybean meal to weaned pigs

This study sought to determine whether fermentation could increase apparent ileal digestibility (AID) of dry matter (DM), nitrogen (N), energy (E) and amino acids (AA) in fermented soybean meal (FSBM) greater than that of soybean meal (SBM) in weaned pigs. Four weaned pigs (10.00 ± 0.30 kg) were surgically equipped with T‐cannulas and randomly followed a 4 × 4 Latin square design of treatments (SBM, FSBM, fish meal and spray‐dried plasma protein). Overall, the fermentation process was able to reduce the amount of anti‐nutritional factors (ANF), including trypsin inhibitors, raffinose and stachyose, in the FSBM diet, which were significantly reduced by 39.4, 92.2, and 92.9%, respectively, as compared to the SBM diet. As a consequence of ANF reduction in FSBM, the AID of DM, N and E as well as AA was significantly greater with FSBM than SBM. Taken all together, the fermentation process improved the nutritional quality of SBM, due to ANF reduction, leading to improvement of digestibility of AA. As such, FSBM can be potentially used as a specialized feed ingredient, especially for young animal diets in an attempt to reduce diet costs.     

黄土高原丘陵区柠条根系生长发育特性研究

以陇西黄土高原丘陵沟壑区1～3a柠条荒山直播苗为研究对象,应用挖掘法对其根系的形态、生长发育、分布及根瘤特性进行了观测分析。结果表明,幼龄期柠条主根生长速度远大于株高生长,根冠比值在4～6之间;侧根主要发生在主根的5～15cm段;柠条胚根平均向上伸长5cm,合理的播种深度为3cm;根瘤主要集中在10—30cm土层中主根及一级侧根上;速生、发达的根系和根瘤,使得柠条具有超强的抗旱、耐瘠薄能力和蓄水保土、改良土壤的生态功能。柠条秋播较春播能更大程度地与当地的光、热、水等资源相拟合,应大力倡导秋季直播造林。     

饲料中添加酵母提取物对凡纳滨对虾免疫相关基因表达及抗菌机能的影响

为了评估饲料中添加酵母提取物对凡纳滨对虾免疫灵敏性和平衡性的相关基因表达的影响,以鱼粉含量为24.5%的基础饲料(对照组)及在基础饲料中添加2.5%酵母提取物的实验饲料,分别饲喂凡纳滨对虾56 d,检测两种饲料投喂的对虾在急性感染溶藻弧菌前后鳃组织Toll受体、IM D和溶菌酶mRNA表达量变化及感染后的对虾死亡情况。结果表明:摄食添加酵母提取物饲料的凡纳滨对虾鳃组织中Toll受体mRNA和溶菌酶mRNA表达量均显著高于对照组对虾(P0.05),IMD mRNA表达量与对照组无显著性差异(P0.05)。急性感染溶藻弧菌后,两组对虾鳃组织Toll受体、IMD和溶菌酶mRNA表达量随感染进程均出现显著变化,Toll受体、IMD和溶菌酶mRNA表达量峰值出现的时间分别为24、42和36 h,且摄食添加酵母提取物组对虾各基因的表达量峰值分别为对照组峰值的201.06%、481.46%和276.77%,均显著高于对照组对虾(P0.05)。摄食添加酵母提取物饲料组对虾鳃组织中Toll受体和IM D mRNA表达量在感染溶藻弧菌后12和24 h分别出现显著上调,而对照组对虾鳃组织中Toll受体和IMD mRNA表达量分别在感染弧菌后24和42 h才出现显著上调。两组对虾经溶藻弧菌人工急性感染后72 h内的累积死亡率无显著差异(P0.05)。实验表明,饲料中添加酵母提取物上调了溶藻弧菌感染前凡纳滨对虾Toll受体和溶菌酶mRNA的表达量,提早了溶藻弧菌感染后对虾Toll受体和IMD mRNA表达量上调时间,且增加了对虾Toll受体、IM D和溶菌酶mRNA表达量的峰值,在一定程度上提高了凡纳滨对虾免疫相关基因表达的灵敏性。     

外源NO对低温胁迫下豇豆幼苗生长和生理特性的影响

研究不同浓度的外源NO供体硝普钠（sodium nitroprusside, SNP）溶液对8 ℃低温胁迫下豇豆幼苗生长和生理特性的影响,结果表明,0.5~1.5 mmol/L外源NO处理下,豇豆幼苗叶面积、根冠比、壮苗指数有所提高,脯氨酸（Pro）、可溶性蛋白和叶绿素含量增加,丙二醛（MDA）含量降低。过高或过低浓度的外源NO对低温胁迫下豇豆幼苗生长的缓解作用均不显著,过低浓度的外源NO会促进MDA累积。在低温胁迫下,适当浓度的外源NO能促进脯氨酸和可溶性蛋白的形成,提高叶绿素含量,降低膜脂过氧化程度,从而增强豇豆幼苗对低温胁迫的适应性。     

华北刺槐林与自然恢复植被土壤微生物量碳、氮含量四季动态

【目的】土壤微生物量碳、氮是植被所需碳、氮的重要“源”或“库”,是公认的综合评价土壤质量或肥力的重要指标,也是土壤生态系统变化的预警及敏感指标,研究其动态变化,可为退耕还林及后期管理决策提供科学依据,并为深入研究林地碳氮循环及温室气体排放提供参考。【方法】以农田( FL)为对照,研究华北土石山区10年生刺槐林、43年生刺槐林、自然恢复植被( NRV)土壤微生物量碳、氮的四季动态变化,并对各样地微生物量碳、氮对土壤营养库的贡献率进行对比研究。【结果】各样地微生物量碳、氮随土层加深而逐渐下降,其季节动态变化差异显著;农田、自然恢复植被、10和43年生刺槐林地0～20 cm 土层微生物量碳、氮含量四季均值分别为251.94,290.68,150.66,197.34 mg·kg －1和30.95,46.46,36.55,45.27 mg·kg －1。其中：自然恢复植被的微生物量碳、氮含量四季均值最高,其微生物量碳含量分别是农田、10和43年生刺槐林的1.15,1.93和1.47倍,微生物量氮含量分别是它们的1.50,1.27和1.03倍;土壤微生物量碳、氮含量随刺槐树龄增大而升高,43年生刺槐林0～20 cm 土层的微生物量碳、氮含量是10年生刺槐林的1.31和1.24倍。各植被样地不同层次土壤微生物量碳氮比季节差异明显,农田、自然恢复植被、10年和43年生刺槐林 0～20 cm 土层碳氮比四季均值分别为8.64,6.26,4.12 和4.36;10,43年生刺槐林碳氮比分别是农田的0.48和0.50倍,是自然恢复植被的0.66和0.70倍。在 0～20 cm 土层中,农田、自然恢复植被、10和43年生刺槐林地微生物量碳对土壤有机碳平均贡献率分别为1.88%,2.00%,1.54%和1.24%,土壤微生物量氮对土壤全氮的平均贡献率分别为1.21%,5.44%,3.55%和2.26%。【结论】各样地土壤微生物量碳、氮之间显著相关,它们与土壤全氮、有机质和速效钾含量均显著相关;除此之外,土壤微生物量碳还与土壤硝态氮含量显著相关。随着树龄的增加刺槐林土壤微生物量尤其是微生物量氮含量显著提高,因而土壤的生物肥力也显著提高;由土壤微生物量碳、氮含量及其对土壤营养库的贡献率可知,自然恢复植被更利于土壤微生物结构、功能的恢复和生物活性的改善。     

Biochar-induced changes in soil properties affected immobilization/mobilization of metals/metalloids in contaminated soils

Purpose

Remediation of metal contaminated soil with biochar is attracting extensive interest in recent years. Understanding the significance of variable biochar properties and soil types helps elucidating the meticulous roles of biochar in immobilizing/mobilizing metals/metalloids in contaminated soils.

Materials and methods

Six biochars were produced from widely available agricultural wastes (i.e., soybean stover, peanut shells and pine needles) at two pyrolysis temperatures of 300 and 700 °C, respectively. The Pb-, Cu-, and Sb-contaminated shooting range soils and Pb-, Zn-, and As-contaminated agricultural soils were amended with the produced biochars. The mobility of metals/metalloids was assessed by the standard batch leaching test, principal component analysis and speciation modeling.

Results and discussion

The changes in soil properties were correlated to feedstock types and pyrolysis temperatures of biochars based on the principal component analysis. Biochars produced at 300 °C were more efficient in decreasing Pb and Cu mobility (>93 %) in alkaline shooting range soil via surface complexation with carboxyl groups and Fe-/Al-minerals of biochars as well as metal-phosphates precipitation. By contrast, biochars produced at 700 °C outperformed their counterparts in decreasing Pb and Zn mobility (100 %) in acidic agricultural soil by metal-hydroxides precipitation due to biochar-induced pH increase. However, Sb and As mobility in both soils was unfavorably increased by biochar amendment, possibly due to the enhanced electrostatic repulsion and competition with phosphate.

Conclusions

It is noteworthy that the application of biochars is not equally effective in immobilizing metals or mobilizing metalloids in different soils. We should apply biochar to multi-metal contaminated soil with great caution and tailor biochar production for achieving desired outcome and avoiding adverse impact on soil ecosystem.

     

开菲尔粒中产球菌样细菌素乳酸菌的分离与鉴定

利用MRS选择培养基筛选开菲尔粒中具有抑菌作用的乳酸菌。采用牛津杯琼脂扩散法,结合对理化抗性特征的鉴定、抗菌谱测定、菌落形态与特征和生理生化实验鉴定,结合伯杰氏手册对分离到的菌株进行鉴定。生理生化反应为乳酸菌,菌落形态与特征为球菌,对多数的革兰氏阴性菌有抑制作用,该菌株为乳酸乳球菌二乙酰乳酸亚种。鉴定分离为产球菌样细菌...