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Rapid detection of Pseudomonas aeruginosa by cross priming amplification
XIANG Yong, YAN Ling, ZHENG Xiao-cui, LI Li-zhen, LIU Peng, CAO Wei-sheng
2020, 19 (10): 2523-2529.   DOI: 10.1016/S2095-3119(20)63187-2
Abstract109)      PDF in ScienceDirect      
Pseudomonas aeruginosa (PA) is an opportunistic pathogen of humans and animals and a common source of nosocomial infections especially of the respiratory tract.  Pseudomonas aeruginosa is also a major bacterial disease of poultry and in particular, eggs and newly hatched chicks.  In this study, we developed a simple, accurate and rapid molecular detection method using cross priming amplification (CPA) with a nucleic acid test strip to detect P. aeruginosa.  The assay efficiently amplified the target gene within 45 min at 62°C only using a simple water bath.  The detection limit of the method was 1.18×102 copies μL–1 for plasmid DNA and 4.4 CFU mL–1 for bacteria in pure culture, and was 100 times more sensitive than conventional PCR.  We screened 83 clinical samples from yellow-feather broiler breeder chickens and hospitalized/treated dogs and cats using CPA, PCR and traditional culture methods.  The positive-sample ratios were 15.3% (13/83) by CPA, 13.3% (11/83) by PCR and 12.1% (10/83) by the culture method.  The established CPA method has significant advantages for detecting P. aeruginosa.  The method is easy to use and possesses high specificity and sensitivity without the requirements of complicated experimental equipment.  The PA-CPA assay is especially fit for outdoor and primary medical units and is an ideal system for the rapid detection and monitoring of P. aeruginosa.
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Aggregate stability and associated C and N in a silty loam soil as affected by organic material inputs
LONG Pan, SUI Peng, GAO Wang-sheng, WANG Bin-bin, HUANG Jian-xiong, YAN Peng, ZOU Juan-xiu, YAN Ling-ling, CHEN Yuan-quan
2015, 14 (4): 774-787.   DOI: 10.1016/S2095-3119(14)60796-6
Abstract2039)      PDF in ScienceDirect      
To make recycling utilization of organic materials produced in various agricultural systems, five kinds of organic materials were applied in a field test, including crop straw (CS), biogas residue (BR), mushroom residue (MR), wine residue (WR), pig manure (PM), with a mineral fertilizer (CF) and a no-fertilizer (CK) treatment as a control. Our objectives were: i) to quantify the effects of organic materials on soil C and N accumulation; ii) to evaluate the effects of organic materials on soil aggregate stability, along with the total organic carbon (TOC), and N in different aggregate fractions; and iii) to assess the relationships among the organic material components, soil C and N, and C, N in aggregate fractions. The trial was conducted in Wuqiao County, Hebei Province, China. The organic materials were incorporated at an equal rate of C, and combined with a mineral fertilizer in amounts of 150 kg N ha-1, 26 kg P ha-1 and 124 kg K ha-1 respectively during each crop season of a wheat-maize rotation system. The inputted C quantity of each organic material treatment was equivalent to the total amount of C contained in the crop straw harvested in CS treatement in the previous season. TOC, N, water-stable aggregates, and aggregate-associated TOC and N were investigated. The results showed that organic material incorporation increased soil aggregation and stabilization. On average, the soil macroaggregate proportion increased by 14%, the microaggregate proportion increased by 3%, and mean-weight diameter (MWD) increased by 20%. TOC content followed the order of PM>WR>MR>BR>CS>CK>CF; N content followed the order WR>PM>MR>BR>CS>CF>CK. No significant correlation was found between TOC, N, and the quality of organic material. Soil silt and clay particles contained the largest part of TOC, whereas the small macroaggregate fraction was the most sensitive to organic materials. Our results indicate that PM and WR exerted better effects on soil C and N accumulation, followed by MR and BR, suggesting that organic materials from ex situ farmland could promote soil quality more as compared to straw returned in situ.
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