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.

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.

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.

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.

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.

水乳型磷酸铝除锈防锈性能及成膜机理研究

研究了水乳型磷酸铝的防锈作用和防锈性能 .根据系统研究结果 ,认为磷酸铝含量大于 4 0 %时 ,防锈膜与基底钢铁制件结合牢固、平整、光洁 。

Studies on Conversion of K in Potash Feldspar into Water-soluble Form

The studies for raising potash feldspar utilization ratio by physical, biological and chemical methods are carried out in laboratory and the results are presented. It is shown that calcinatory and biological methods have positive effects on conversion of K in potash feldspar into water-soluble form, but the conversion is low; chemical method can change most of insoluble potassium into available form, and might be significant in industry to some extent.

Study on Piezoelectric Immunosensor for the Detection of H9-subtype Avian Influenza Virus

[Objective] The aim is to develop the piezoelectric immunosensor to detect H9-subtype avian influenza virus（AIV）.[Method] The immunosensor chip was constructed by self-assembling mercaptopmpionic acid（MPA） to be monolayer on the silver-coated electrode of quartz crystal and coupling the monoclonal antibody to H9 subtype AIV with N-ethy-N＇-（3-dimethyl aminopropyl）carbodiimide hydrochloride（EDC） and N-hydroxysuccinimide（NHS）.The immunosensor to detect H9 subtype AIV was established.[Result] The results showed that the immunosensor displayed better specificity to H9 AIV and had no response to H5AIV and NDV when it was used for detection.The sensitivity test indicated the detection sensitivity for the H9 subtype AIV could reach 20-100 EID50.[Conclusion] The research provided a foundation for further research on the immunosensor for detecting AIV and it could be a new approach to detect other related viruses.

Biodegradation of Lubricating Oil in Wastewater with Zoogloea sp.

The purpose of this study was to identify microbial strains that have a strong ability to biodegrade lubricating oil. No. 20 lubricating oil was used as the sole carbon source in an isolation medium to screen bacteria from sludge that had been contaminated with crude oil. Through both morphological and biochemical methods, the bacterial strain that had the highest biodegrading capacity was identified as Zoogloea sp. On the basis of these preliminary results, a biological contact oxidation method was employed to further assess the ability of the Zoogloea sp. strain to treat wastewater contaminated by No. 20 lubricating oil using three oxidation cabins and two hydraulic retaining times （HRT）. Results showed that the concentration of No. 20 lubricating oil with a 12 h hydraulic retaining time and 16.5 L h^-1 inflow rate was reduced by 99.3% in 15 d, and with a 6 h hydraulic retaining time and 33 L h%^-1 inflow rate it decreased by 98.6070 in 12 d. The results from this study indicated that Zoogloea sp. had a strong potential to be utilized in biodegradation of lubricating oil.

Quality-control method for the determination of biological activity of engineered calcineurin subunit B

The aim of this study was to establish a quality-control method for calcineurin subunit B(CNB) biological activity determinations. CNB enhances the p-nitrophenylphosphate(p NPP) dephosphorylating activity of calcineurin subunit A Δ316 mutant(CNAΔ316). A series of CNB concentrations were fitted to a four-parameter equation to calculate the corresponding p NPP maximum dephosphorylation rates. Values were calculated based on biological activity references using a parallel line method. The method was then validated for accuracy, precision, linearity, linear range, sensitivity, specificity, and robustness. The recovery results were greater than 98%. Intra-plate precision was 6.7%, with inter-plate precision of 10.8%. The coefficient of determination was greater than 0.98. The linear range was 0.05–50 μg m L?1, with sensitivity of 50 μg m L?1. Tested cytokines did not induce CNAΔ316 dephosphorylation of p NPP. The chosen CNAΔ316 concentration range did not affect activity determinations.

Development and application of biological technologies in fish genetic breeding

Fish genetic breeding is a process that remolds heritable traits to obtain neotype and improved varieties.For the purpose of genetic improvement,researchers can select for desirable genetic traits,integrate a suite of traits from different donors,or alter the innate genetic traits of a species.These improved varieties have,in many cases,facilitated the development of the aquaculture industry by lowering costs and increasing both quality and yield.In this review,we present the pertinent literatures and summarize the biological bases and application of selection breeding technologies(containing traditional selective breeding,molecular marker-assisted breeding,genome-wide selective breeding and breeding by controlling single-sex groups),integration breeding technologies(containing cross breeding,nuclear transplantation,germline stem cells and germ cells transplantation,artificial gynogenesis,artificial androgenesis and polyploid breeding)and modification breeding technologies(represented by transgenic breeding)in fish genetic breeding.Additionally,we discuss the progress our laboratory has made in the field of chromosomal ploidy breeding of fish,including distant hybridization,gynogenesis,and androgenesis.Finally,we systematically summarize the research status and known problems associated with each technology.

Ecological restoration of small water courses,experiences from Germany and from projects in Beijing

From 2009 until 2012 the project“Watershed Management of Forest Land in Beijing,Restoration of Small Water Bodies(SWBR)”was implemented,combining Close to Nature Forest Management and Restoration of Small Water Bodies.The targets were to improve flood control,to enhance the ecological conditions by copying nature and to support the recreational value of small water bodies,all in cooperation with people living there.The efficiency of each project was proofed by comparison of biological and hydro-morphological assessment before the projects started and 2-3 years after they were finished.The results confirmed the ecological improvements of the restored river sections and showed the achievements.Guidelines to assess the biological and hydro-morphological status of rivers were developed and there are plans to introduce them as Beijing Standards.Planning and implementation of measures,based on experiences in Central Europe,will be documented in a handbook.&2015 International Research and Training Center on Erosion and Sedimentation and China Water and Power Press.Production and Hosting by Elsevier B.V.This is an open access article under the CC BY-NC-ND license(http://creativecommons.org/licenses/by-nc-nd/4.0/).