Screening of Preprocessing Method of Biolog for Soil Microbial Community Functional Diversity

As one of the main methods of microbial community functional diversity measurement, biolog method was favored by many researchers for its simple oper- ation, high sensitivity, strong resolution and rich data. But the preprocessing meth- ods reported in the literatures were not the same. In order to screen the best pre- processing method, this paper took three typical treatments to explore the effect of different preprocessing methods on soil microbial community functional diversity. The results showed that, method B＇s overall trend of AWCD values was better than A and C＇s. Method B＇s microbial utilization of six carbon sources was higher, and the result was relatively stable. The Simpson index, Shannon richness index and Car- bon source utilization richness index of the two treatments were B〉C〉A, while the Mclntosh index and Shannon evenness were not very stable, but the difference of variance analysis was not significant, and the method B was always with a smallest variance. Method B＇s principal component analysis was better than A and C＇s. In a word, the method using 250 r/min shaking for 30 minutes and cultivating at 28 ℃ was the best one, because it was simple, convenient, and with good repeatability.

Establishment of Molecular Biological Method for Identification of Bacteria by 16S rDNA and gyrB Gene

[Objectives]The paper was to establish a molecular biological method for identification of bacterial strains.[Methods]The thalli of standard bacterial strains existing in the laboratory were collected and genomic DNA was extracted for amplification of 16S rDNA and gyrB gene.The 16S rDNA and gyrB gene sequences were obtained after sequencing.Sequences were aligned and analyzed via EzBioCloud and NCBI database,and phylogenetic trees were constructed to determine the species relationship of strains.Meantime,they were compared with known strains.[Results]This method could identify 5 standard strains accurately to the species level.The 16S rDNA and gyrB gene sequences were aligned and analyzed in EzBioCloud database and NCBI database.The strain with the max score was consistent with the known strain.And the query cover and ident were both above 99%.[Conclusions]The established molecular biological method for identification of bacterial strains by 16S rDNA and gyrB gene has good accuracy,which effectively solves the problem that the laboratory identification of bacteria relies on traditional methods and the accuracy can not be guaranteed,and further improves the identification ability of laboratory bacterial strains.

Analytical methods for determination of terbinafine hydrochloride in pharmaceuticals and biological materials

Terbinafine is a new powerful antifungal agent indicated for both oral and topical treatment of myco- sessince. It is highly effective in the treatment of determatomycoses. The chemical and pharmaceutical analysis of the drug requires effective analytical methods for quality control and pharmacodynamic and pharmacokinetic studies. Ever since it was introduced as an effective antifungal agent, many methods have been developed and validated for its assay in pharmaceuticals and biological materials. This article reviews the various methods reported during the last 25 years.

Estimation of the Biological Methods of Assessing Soil N-Supplying Capacity in Calcareous Soil

Although many biological methods are used to determine soil nitrogen supplying capacity, there are certain differences in the results for different types of soils and various ways of measurement due to the complexity of soil N conformation, the high variance of soil and microorganism, and the difference of environment. Therefore, it is not clear about which biologic incubation method is better for calcareous soil. In this study, pot experiments were performed by using 25 different calcareous surface soil samples on the Loess Plateau and taking the N uptake of wheat and corn with leaching soil initial nitrate and without leaching in pot experiments as the control to investigate the difference of eight biological incubation methods for reflecting soil nitrogen supply capacity. The eight biological methods are waterlogged incubation, aerobic incubation for 2 weeks and for 4 weeks, dry-wet alternation aerobic incubation for 2 weeks, long-term alternate leaching aerobic incubation （and N mineralization potential, No）, short-term leaching aerobic incubation, microbial biomass carbon （Bc）, and microbial biomass nitrogen （BN） method, respectively. Among these methods, the dry-wet alternation aerobic incubation and aerobic incubation for 4 weeks were the modification of the method of aerobic incubation for 2 weeks according to the actual farmland moisture. The results showed that the correlation coefficients between these methods and crop uptake N with leaching soil initial nitrate were 0.530, 0.700, 0.777, 0.768, 0.764 （and 0.790, No）, 0.650, 0.555, and 0.465, respectively （r0.05 = 0.369, r0.0l = 0.505）. While without leaching soil initial nitrate, their coefficients were 0.351, 0.963, 0.962, 0.959, 0.825 （and0.812, No）, 0.963, 0.289, and 0.095, respectively （r0.05 = 0.369, r0.01 = 0.505）. In conclusion, excluding the soil initial nitrate, the correlation coefficients between the eight methods and crop uptake N were, from high to low, N0, aerobic incubation for 4 weeks, dry-wet alternation aerobic incubation for 2 weeks, and long-term alternate leaching aerobic incubation, while including the soil initial nitrate the correlation coefficients between them increased significantly and the values were all beyond 0.950 for these four methods, including aerobic incubation for 2 weeks and for 4 weeks, dry-wet alternation aerobic incubation for 2 weeks and short-term leaching aerobic incubation. The waterlogged incubation method, Bc and BN in the calcareous soil, had lower correlation coefficient with crop uptake nitrogen compared with other methods. Thus, dry-wet alternation aerobic incubation for 2 weeks was a better index for evaluating calcareous soil N supply capacity due to some other methods having disadvantages and not suitable for the actual farmland characteristics.

Assessment of Domestic Water Sources Safety: Application of the Micro Biological Survey Method and Microbiological Profile of the Contaminating Bacteria

Water is essential for human life and it’s expected to be clean and safe. In Cameroon, government efforts to improve access to drinking water are mitigated. Many households are suffering from chronic water shortages leading to the majority of people using doubtful water sources. This study was carried out in accordance with guidelines produced by the STARBIOS2-EU funded project on Responsible Research and Innovation and aimed to assess the microbiological quality of water samples collected from domestic water sources in 3 divisions of the West region of Cameroon. 22 water samples from well water, stream, water pump and river were aseptically collected. At each sampling point, 50 mL of sample was taken aseptically and immediately transported to the laboratory for analysis. The detection of Coliforms bacteria was done using Micro Biological Survey (MBS) method. 1 mL of each sample was inoculated in the Coliforms MBS (Coli MBS) vial initially rehydrated with 10 mL of sterile distilled water. The Coli MBS vials were closed, shaken for homogenization, and then incubated at 37°C. From the initial red color of the Coli MBS vials, changes in color to yellow of the reaction vials were monitored at three different time intervals (12 h, 19 h and 24 h), corresponding to three levels of bacterial concentration. All positives samples on MBS method were selected to determine the presence of E. coli Loop full broths of Coli MBS vials were taken;streaked into the eosin methylene blue (EMB) agar plate and incubated at 37°C for 24 hours. E. coli was confirmed by observing green metallic sheen on EMB agar plate. The biochemical indole, methyl red, Voges-Proskauer and citrate (IMViC) test was performed to determine the complete microbiological profile of the water samples. Almost all the water samples were contaminated with Total Coliforms (TC). A high concentration of TC (>103 CFU/mL) was found in 8 samples (36.4%), a medium concentration (10 < x < 103 CFU/mL) was observed in 10 samples (45.5%) and 2 samples (9.1%) were low concentrated (1 < x < 10 CFU/mL). 8 samples were positive for the presence of E. coli by observing the green metallic sheen on EMB agar plates. The IMViC test confirmed the presences of 5 bacteria species: Enterobacter spp., Klebsiella spp., E. coli spp., Salmonella spp. and Shigella spp. We recommend integrating sanitary assessment in the design and implementation of water supply projects. Regular water purification using proper methods and maintenances of the water point. Regular microbiological quality assessment of water intended for human consumption should be planned and carried out.

Prevention and Control Effects of New Biological Formulations Sprayed by Different Methods on Proceras venosatus (Walker) and Tryporyza intacta Snellen in Middle and Late Stages

To screen new biological formulations, accurate and efficient application technology, field test was conducted with 72% Bacillus thuringiensis(Bt)·monosultap WP, 8% lambda-cyhalothrin·emamectin benzoate SA and 3.6% lambda-cyhalothrin·Bt SA through manual spraying and unmanned aerial vehicle spraying. Test results and comprehensive evaluation analysis demonstrated that 72% Bt·monosultap WP and 8%lambda-cyhalothrin·emamectin benzoate SA by manual spraying and unmanned aerial vehicle spraying had good prevention and control effect on strains and internodes damaged by Tryporyza intacta Snellen in middle and late stage, which were ideal new biological formulations with high effi-ciency and low risk for prevention and control of T. intacta in middle and late stage, and could be popularized in sugarcane area. 72% Bt·monosultap WP at the dose of 3 000 g/hectare and 8% lambda-cyhalothrin·emamectin benzoate SA at the dose of 750 mL/hectare could be sprayed in mid September at the peak occurrence period of the forth and fifth generations of T. intacta and Proceras venosatus. Agents were diluted with 900 kg water per hectare, and manually sprayed with electric knapsack sprayer; or agents were diluted with 15 kg of Haoyang aerial control special addi-tives and water per hectare, and sprayed with unmanned aerial vehicle. The control effect against borer-damaged strain rate was above 81.3% and that against borer-damaged internode rate was above 88.6%.

采用Biolog法分析制药废水处理工艺中微生物多样性

采用水解酸化池、活性污泥池、曝气生物滤池组合工艺处理东北制药总厂的制药废水,在污泥驯化成功并运行稳定后,对COD和BOD5的去除率均在90%以上,出水水质达到了《污水综合排放标准》(GB 8978—1996)的二级标准。采用Biolog方法分析了接种污泥及各构筑物中微生物群落的多样性,结果表明,接种污泥和水解酸化池污泥的停滞期比活性污泥池污泥和曝气生物滤池生物膜的长,但各微生物群落在稳定期的平均活性相差不大。多样性指数分析结果表明,各构筑物中微生物群落的丰富度和均一性相近,但最常见的物种不同;主成分分析结果显示,3种污泥及生物膜对ECO板上碳源的利用情况存在差异。

添加菌剂对堆肥化过程中微生物群落代谢影响的Biolog解析

为了促进堆肥中木质纤维素降解并了解其中微生物群落代谢功能多样性,该文主要采用Biolog微平板法,考察了添加菌剂对牛粪堆肥中木质纤维素降解及微生物群落代谢能力的影响。结果表明:堆肥结束时,菌剂处理的堆肥中纤维素、半纤维素和木质素的降解率分别比对照提高了31.31%、19.57%和14.33%;对平均色度变化分析表明接种菌剂增加了堆肥高温期微生物的细胞代谢能力;Shannon指数、Simpson指数和McIntosh指数分析均显示添加菌剂可以显著提高堆肥前期的微生物多样性;主成分分析结果显示,添加菌剂提高了堆体中微生物对双亲化合物、聚合物、氨基酸和氨基化合物等碳源的代谢能力,从而可促使有机质被更高效降解。

Biolog法测定土壤微生物群落功能多样性预处理方法的筛选

作为测定微生物群落功能多样性主要方法之一的biolog法,由于操作简单、灵敏度高、分辨力强、获得数据量丰富,受到众多研究者的青睐,但文献中报道的有关实验预处理方法却不尽相同。为了筛选最佳的预处理方法,通过比较文献中3种较为常用的方法,研究不同预处理对biolog法测定微生物群落功能多样性的影响。结果表明:AWCD值总体变化趋势B方法优于A、C;微生物对六类碳源的利用率中B方法对大部分碳源利用较高,且结果较稳定;两种处理的Simpson指数、Shannon丰富度指数及碳源利用丰富度指数均B>C>A,而Mc Intosh指数、Shannon均匀度结果不太稳定,但经方差分析,其差异不显著,且B方法的方差最小;主成分分析的结果 B方法优于A与C。总之,在250 r/min的转速下震荡30 min,并于28℃条件下培养接种平板的处理(B法)效果较好,其测定结果重现性好,且操作简便快捷,是比较理想的预处理方法。

Biolog ECO分析活性污泥微生物功能多样性特征

利用Biolog ECO技术对余杭污水处理厂污泥原位减量示范工程的活性污泥微生物多样性进行分析。结果表明:实验组利用全部(6类)碳源能力得到增强,群落的丰富度、均匀度和多样性具有显著提高;实验组对酚酸类、胺类碳源的利用度远高于对照组;其中对α-D-乳糖、L-苯基丙氨酸、α-环式糊精、4-羟基苯甲酸、苯乙胺等5种碳源的利用能力显著提高。综合分析表明,菌剂强化后可使活性污泥微生物群落得到优化,使得生化系统对碳源利用的深度和广度均有较大提高。

采用Biolog法分析餐厨垃圾厌氧消化微生物群落多样性

该研究在中温条件下进行实验室规模的连续式餐厨垃圾厌氧消化试验,用Biolog方法分析了反应器各个运行阶段污泥中微生物群落的多样性。多样性指数分析结果表明:反应器中各个阶段微生物物种丰富度和常见的物种接近,但均一性各阶段有较大的差异。ECO板上碳源的利用情况不同,表明反应器各个阶段微生物群落呈现出不同的微生物多样性。主成分分析结果显示,反应器在启动与稳定运行阶段微生物种群多样性较为接近,而与反应器抑制阶段和恢复阶段微生物种群多样性差异性明显。

养殖池塘底泥预处理方法对Biolog Eco板分析结果的影响

为探讨不同预处理方法对养殖池塘底泥Biolog Eco板分析结果的影响,以南美白对虾养殖池塘周边底泥1#和中部底泥2#为研究对象,考察了文献中常用的5种预处理方法底泥微生物群落平均颜色变化率( Average well color development,AWCD)、碳源利用模式和多样性指数的差异。结果表明,培养3 d后周边底泥1#样品异养细菌总数(7.73×10^5 CFU·mL^-1)显著低于中部底泥2#样品(3.48×10^7 CFU·mL^-1)(P<0.05)。不同预处理方式底泥微生物群落的AWCD值、六类碳源利用情况、多样性指数及主成分分析差异较大。240 h时,1#底泥方法4和方法5除对胺类碳源的利用(方法5显著高于方法4)外,微生物群落的上述其他指标差异均不显著(P>0.05),但有多项指标显著优于方法1、2、3(P<0.05);2#底泥方法5处理微生物群落的多项指标显著优于方法1~4;且无论在1#底泥还是2#底泥,方法5主成分分析的平行点均较为集中,说明其平行性较好。因此建议采用方法5进行养殖池塘底泥的预处理,即10%底泥稀释液在250 r·min^-1转速下振荡30 min,经600 r·min^-1离心10 min,离心上清液直接接入Biolog Eco板。

利用BIOLOG系统对不同种类细菌鉴定的研究

为完善细菌鉴定工作,本试验通过利用B IOLOG细菌自动微生物分析系统(简称B I-OLOG系统)对13株大肠杆菌、15株巴氏杆菌和7株金黄色葡萄球菌进行了鉴定,并与传统方法[1]包括形态学观察、革兰氏染色、生化实验等的鉴定结果进行了比对。实验结果表明,与传统方法相比B IOLOG系统具有准确、快捷、简便的优点,同时也有一定的局限性,从而为B IOLOG系统的更好使用提供了实验依据。

Wet reclamation of sodium silicate used sand and biological treatment of its wastewater by Nitzschia palea

The massive amount of sodium silicate in the used sand was a pollution source, especially in the waste water from the wet reclamation of used sand. A new process of wet reclamation by biologically treating the waste water produced during the wet reclamation process of used sand was studied in the paper. In the work, the precultivation of N. palea was performed firstly, and three different scrubbing solutions： （1） tap water, （2） modified medium for N. palea, and （3） filtrate of the broth treated by N. palea for 15 days, were used. The results of the primary investigation show that a de-skinning ratio of 90% is obtained when using the scrubbing solution containing modified medium for N. palea at the ratio 1：2 of sand and scrubbing solution, and the maximal concentrations of Na^＋ and SiO3^2- are 1.49 g.L-1 and 0.51 g.L-1, respectively. The results of the optimal biomass, pH value decrease and Na^＋ and SiO32 consumption indicate the optimal incubation conditions are at the irradiance of 5,000 lux and 25 ℃. Using the filtrate of the broth treated by N. palea for 15 days as the scrubbing solution directly, a de-skinning ratio of 93% is the highest compared to the results of the tap water and the modified medium for N. palea. In the biological process using N. palea, less water is used and little wastewater is produced, which is advantageous to the purpose of green manufacturing and environmental protection.

Biolog-ECO方法探究饲喂益生菌对凡纳滨对虾肠道微生物代谢及有效作用时间的影响

以霍氏肠杆菌(E3)和乳酸菌(R3)2株益生菌对凡纳滨对虾进行为期4周的养殖饲喂实验,饲喂后期利用Biolog-ECO方法对实验组及空白组的凡纳滨对虾肠道微生物菌群多样性的差异进行比较分析,以评价益生菌对凡纳滨对虾肠道微生物菌群代谢功能的影响。结果显示,添加霍氏肠杆菌(E3)或乳酸菌(R3)的实验组,与空白组相比较,平均每孔颜色变化率显著上升,表明益生菌增强了肠道微生物活性;凡纳滨对虾肠道微生物利用各类碳源的整体能力显著增强,表明益生菌可以促进水产动物的代谢功能;肠道微生物多样性指数(包括Shannon、Simpson和McIntosh指数)有明显差异,表明饲喂2株益生菌可以提高凡纳滨对虾肠道菌群的丰富度。其中,停喂霍氏肠杆菌后第1天和第5天取样结果表明,Shannon指数显著降低,Simpson和McIntosh指数显著升高;停喂乳酸菌后的第1天和第5天取样结果表明,Shannon指数无显著差异,Simpson和McIntosh指数显著升高;二者在第10天取样的结果中均无显著差异,表明饲料中添加益生菌可以改变凡纳滨对虾肠道内原有菌群的数量和结构,促进对虾肠道内微生物群落间复杂的相互作用,进而在维持或者促进对虾健康方面发挥着重要的作用,同时也表明此两株益生菌在凡纳滨对虾肠道中停留时间最少为5 d。

Biologically Inspired Design of Context-Aware Smart Products

The rapid development of information and communication technologies(ICTs)and cyber-physical systems(CPSs)has paved the way for the increasing popularity of smart products.Context-awareness is an important facet of product smartness.Unlike artifacts,various bio-systems are naturally characterized by their extraordinary context-awareness.Biologically inspired design(BID)is one of the most commonly employed design strategies.However,few studies have examined the BID of context-aware smart products to date.This paper presents a structured design framework to support the BID of context-aware smart products.The meaning of context-awareness is defined from the perspective of product design.The framework is developed based on the theoretical foundations of the situated function-behavior-structure ontology.A structured design process is prescribed to leverage various biological inspirations in order to support different conceptual design activities,such as problem formulation,structure reformulation,behavior reformulation,and function reformulation.Some existing design methods and emerging design tools are incorporated into the framework.A case study is presented to showcase how this framework can be followed to redesign a robot vacuum cleaner and make it more context-aware.

Advances in Studies of Biological Nitrogen Saving Technology for Wheat

This paper first described the current status of wheat nitrogen fertilizers utilization rate. Then,from balanced fertilizer application,applying slow/controlled release fertilizer,deep application of nitrogen fertilizers and graded application,controlled application amount,selection of application time according to soil properties,diagnosed application of fertilizer with instrument,addition of synergist,and precise fertilizer application,it summarized biological nitrogen saving methods and introduced prospects of future biological nitrogen saving technology.

A Direct Droplet Digital PCR Method for <i>E. coli</i>Host Residual DNA Quantification

Injectable drugs manufactured in E. coli must be tested for host residual DNA (hr DNA) impurity in ensuring drug purity and safety. Because of low allowable hr DNA as impurity, highly sensitive methods are needed. Droplet digital PCR (ddPCR) is a new method where the reaction is partitioned into about 20,000 nanoliter-sized droplets and each droplet acts as individual PCR reaction. After completion of end-point PCR, droplets are analyzed for fluorescence and categorized as positive or negative and DNA quantified using Poisson statistics. Here we describe development of a direct E. coli hr DNA dd PCR method where the drug is directly added to the ddPCR reaction. We show that the ddPCR method has acceptable precision and high accuracy, works with different biologic drugs, and compared to qPCR shows higher tolerance of drug matrices. The method does not require DNA extraction or standard curves for quantification of hr DNA in unknown samples.

COMPUTATION OF EM FIELD DISTRIBUTION IN A BIOLOGICAL MEDIUM AND EXPERIMENTAL CONFIRMATION

Theory and numerical computation of EM field distribution rule in a biologicalmedium with irregular shape are described. Integral equations of induced EM field distributionin biological body are deduced from EM field scattering. Numerical computations are conductedby the moment method. In order to verify the effectiveness of the theory, a set of experimentalequipment is designed precisely. The results of numerical computation are compared with mea-sured values. It is shown that they are in good agreement. This theory can be used widely forits generality.

Syntheses of Fe-Ni Magnetic Nanoparticles and Their Applications on Biologic Labeling

In this study,we compared FeNi alloy magnetic nanoparticles(MNPs) prepared by either combustion or chemical precipitation methods.We found that the FeNi MNPs generated by combustion method have a rather high saturation magnetization Ms of ～180 emu/g and a coercivity field Hc of near zero.However,the alloy nanoparticles are easily aggregated and are not well dispersive such that size distribution of the nanoparticle clusters is wide and clusters are rather big(around 50～700 nm).To prepare a better quality and well dispersed Fe-Ni MNPs,we also developed a thermal reflux chemical precipitation method to synthesize FeNi3 alloy MNPs.The precursor chemicals of Fe(acac)3 and Ni(acac)2 in a molecular ratio of 1:3 reacted in octyl ether solvent at the boiling point of solvent(～300 ℃) by the thermal reflux process.The 1,2-hexadecandiol and tri-n-octylphosphine oxide(TOPO) were used as reducer and surfactant,respectively.The chemically precipitated FeNi3 MNPs are well dispersed and have well-controlled particle sizes around 10～20 nm with a very narrow size distribution(±1.2 nm).The highly monodispersive FeNi3 MNPs present good uniformity in particle shape and crystallinity on particle surfaces.The MNPs exhibit well soft magnetism with saturation magnetization of ～61 emu/g and Hc～0.The biomedically compatible FeNi MNPs which were coated with biocompatible polyethyleneimine(PEI) polymer were also synthesized.We demonstrated that the PEI coated FeNi MNPs can enter the mammalian cells in vitro and can be used as a magnetic resonance imagine(MRI) contrast agent.The results demonstrated that FeNi MNPs potentially could be applied in the biomedical field.The functionalized magnetic beads with biocompatible polymer coated on MNPs are also completed for biomedical applications.