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The occurrence, distribution and removal efficiencies of organophosphorus flame retardants (OPFRs) and metals were examined in a municipal landfill leachate treatment system in Guangzhou, China. Five OPFRs and thirty-five metals were detected in wastewater samples collected at different treatment stages. ∑OPFRs was reduced from 4807.02 ng L−1 to 103.91 ng L−1 through the treatment system, with close to 98% removed from the dissolved phase. Tris(clorisopropyl) phosphates (TCPPs) dominated through the treatment process and accounted for over 80% and 50% of ∑OPFRs at the influent and the effluent, respectively. TCPPs were most efficiently removed (98.6%) followed by tris(2-chloroethyl) phosphate (TCEP) (96.6%) and triphenyl phosphate (TPP) (88.5%). For metals, Fe, Cr, and Rb were dominant in the raw leachate, detected at 7.55, 2.82, and 4.50 mg L−1, respectively. Thirteen regulated heavy metals – including eight major pollutants (i.e., As. Cd, Cr, Cu, Hg, Ni, Pb, and Zn) – have been detected in all wastewater samples at sub-mg L−1 levels. Over 99.5% removal was achieved for Cr, Ni, and Fe, and close to 95% removal efficiency was observed for Rb. For the eight major heavy metals, over 99% removal was observed; the only exception was Cu, which was removed at 89%. It was found that microfiltration/reverse osmosis was critical for the removal of OPFRs and heavy metals while the core biological treatment played a minor role towards their removal. Remobilization of Co, Cu, Fe, Hg, Mn, Ni, Sb, and Sr from the returned sludge occurred during the second denitrification, indicating the need for additional post-biological process for effective removal of both contaminants. This study highlights the critical need to develop cheap, effective treatment technologies for contaminants-laden leachate generated from open dumps and under-designed landfills.

The YG-13A mission, launched in late 2015, is the first Chinese synthetic aperture radar (SAR) satellite to achieve sub-meter resolution. In this paper, we briefly introduce this milestone satellit...

Yaogan-5(YG-5), launched in December 2008, is a Chinese high-resolution spaceborne synthetic aperture radar (SAR) satellite, with a ground resolution of 3 m. However, the direct geometric positioni...

YaoGan-18 (YG-18) is a Chinese high-resolution synthetic aperture radar (SAR) satellite, which was launched on 29th October 2013. So far, however, there have been no published reports evaluating the geometric accuracy of its imagery; this is rectified in this paper. A refined rational function model (RFM) with an affine transformation is utilised for the spaceborne SAR processing. By setting corner reflectors as ground control points (GCPs) and independent check points, the orthorectification accuracy of YG-18 satellite images was better than 2·5 m. Its stereo-SAR measurement accuracy was subsequently evaluated using a high-precision digital surface model (DSM), achieving an elevation accuracy of 11·36 m. The feasibility of the orthorectification method based on GCPs selected from corresponding image points of YG-18 SAR images and ZiYuan-3 (ZY-3) optical images was also verified, offering single-image accuracy better than 4·5 m.

Geometric calibration is an important means of improving the absolute positioning accuracy of space-borne synthetic aperture radar imagery. The conventional calibration method is based on a calibration field, which is simple and convenient, but requires a great deal of manpower and material resources to obtain ground control points. Although newer cross-calibration methods do not require ground control points, calibration accuracy still depends on a periodically updated reference image. Accordingly, this study proposes a geometric self-calibration method based on the positioning consistency constraint of conjugate image points to provide rapid and accurate calibration of the YaoGan-13 satellite. The proposed method can accurately calibrate geometric parameters without requiring ground control points or high-precision reference images. To verify the absolute positioning accuracy obtained using the proposed self-calibration method, YaoGan-13 Stripmap images of multiple regions were collected and evaluated. The results indicate that high-accuracy absolute positioning can be achieved with a plane accuracy of 3.83 m or better for Stripmap data, without regarding elevation error. Compared to the conventional calibration method using high-accuracy control data, the difference between the two methods is only about 2.53 m, less than the 3-m resolution of the image, verifying the effectiveness of the proposed self-calibration method.

Every year, there are a large number of people died in traffic accidents in our country. If people can get help in time after a traffic accident, the survival rate of the people who are treated can be greatly increased. Therefore, it is very necessary to have a system that can detect traffic accidents and call the police or notify the rescuers in time. According to the survey and analysis, the functions of some similar systems in China are not perfect and most of the cost are high, which cannot be widely promoted in China with a high private car base. Based on the acceleration sequence value obtained by the smartphone accelerometer, this project designs the vehicle collision detection algorithm and realizes the collision detection during the driving process of the vehicle in the android environment. When a collision event is detected, the system can promptly send the collision information to the preset phone number and drive the camera function of the mobile phone, perform instant photo forensics on the scene, and finally send the help information and the accident photo to the preset phone number. The project system can realize the detection and alarm function of automobile collision accidents in time, and provide timely rescue after the collision accident.

It is a problem that most traffic scholars have been studying to enable motor vehicles to pass through signalized intersections without parking or with little parking, so as to improve the traffic efficiency of urban road network and reduce congestion. In the vehicle-road collaboration environment, speed guidance strategy has become an effective solution and trend, aiming at the characteristics and shortcomings of the current trunk signal coordination control as a passive response control based on vehicle arrival drive. In this paper, the principle of speed guidance strategy at signalized intersections is systematically proposed, and the corresponding speed guidance strategy model is proposed according to the different traffic conditions when the guided vehicles arrive at the intersections. This strategy can make the guided vehicles pass through the intersections without stopping or stopping time at least, reduce the delay of vehicles and improve the traffic efficiency at the intersections.

As an effective means of traffic organization, one-way traffic plays an important role in easing traffic congestion. However, the reality is often a one-way traffic organization based on experience, and new traffic problems have emerged during the implementation process. Based on the discussion of one-way traffic setting conditions, this paper models and evaluates the one-way traffic organization scheme that meets the conditions. The fuzzy distribution-analytic hierarchy process is used to evaluate the traffic patency, safety, economy, comfort and environmental impact of the organization scheme. Then, the setting condition-evaluation method is applied to the actual case in combination with VISSIM simulation for inspection. Finally, the experimental results show that the setting conditions-evaluation method can provide a scientific basis for the design and comparison process of one-way traffic organization.

Numerous countries have established their own Earth observing systems (EOSs) for global change research. Data acquisition efforts are generally only concerned with the completion of the mission regardless of the potential to expand into other areas, which reduces the application effectiveness of Earth observation data. This paper explores the cartographic possibility of images being not initially intended for surveying and mapping, and a novel method is proposed to improve the geometric performance. First, the rigorous sensor model (RSM) is recovered from the rational function model (RFM), and then the system errors of the non-cartographic satellite&rsquo, s imagery are compensated by using the conventional geometric calibration method based on RSM, finally, a new and improved RFM is generated. The advantage of the method over traditional ones is that it divides the errors into static errors and non-static errors for each image during the improvement process. Experiments using images collected with the Gaofen-1 (GF-1) wide-field view (WFV) camera demonstrate that the orientation accuracy of the proposed method is within 1 pixel for both calibration and validation images, and the obvious high-order system errors are eliminated. Moreover, a block adjustment test shows that the vertical accuracy is improved from 21 m to 11 m with four ground control points (GCPs) after compensation, which can fulfill requirements for 1:100,000 stereo mapping in mountainous areas. Generally, the proposed method can effectively improve the geometric potential for images captured by non-cartographic satellite.

To better reduce image speckle noise while also maintaining edge information in synthetic aperture radar (SAR) images, we propose a novel anisotropic diffusion algorithm using weighted Euclidean distance (WEDAD). Presented here is a modified speckle reducing anisotropic diffusion (SRAD) method, which constructs a new edge detection operator using weighted Euclidean distances. The new edge detection operator can adaptively distinguish between homogenous and heterogeneous image regions, effectively generate anisotropic diffusion coefficients for each image pixel, and filter each pixel at different scales. Additionally, the effects of two different weighting methods (Gaussian weighting and non-linear weighting) of de-noising were analyzed. The effect of different adjustment coefficient settings on speckle suppression was also explored. A series of experiments were conducted using an added noise image, GF-3 SAR image, and YG-29 SAR image. The experimental results demonstrate that the proposed method can not only significantly suppress speckle, thus improving the visual effects, but also better preserve the edge information of images.

The Chinese Gaofen-3 (GF-3) mission was launched in August 2016, equipped with a full polarimetric synthetic aperture radar (SAR) sensor in the C-band, with a resolution of up to 1 m. The absolute positioning accuracy of GF-3 is of great importance, and in-orbit geometric calibration is a key technology for improving absolute positioning accuracy. Conventional geometric calibration is used to accurately calibrate the geometric calibration parameters of the image (internal delay and azimuth shifts) using high-precision ground control data, which are highly dependent on the control data of the calibration field, but it remains costly and labor-intensive to monitor changes in GF-3’s geometric calibration parameters. Based on the positioning consistency constraint of the conjugate points, this study presents a geometric cross-calibration method for the rapid and accurate calibration of GF-3. The proposed method can accurately calibrate geometric calibration parameters without using corner reflectors and high-precision digital elevation models, thus improving absolute positioning accuracy of the GF-3 image. GF-3 images from multiple regions were collected to verify the absolute positioning accuracy after cross-calibration. The results show that this method can achieve a calibration accuracy as high as that achieved by the conventional field calibration method.

The GF-3 satellite is the first multi-polarization synthetic aperture radar (SAR) imaging satellite in China, which operates in the C band with a resolution of 1 m. Although the SAR satellite system was geometrically calibrated during the in-orbit commissioning phase, there are still some system errors that affect its geometric positioning accuracy. In this study, these errors are classified into three categories: fixed system error, time-varying system error, and random error. Using a multimode hybrid geometric calibration of spaceborne SAR, and considering the atmospheric propagation delay, all system errors can be effectively corrected through high-precision ground control points and global atmospheric reference data. The geometric calibration experiments and accuracy evaluation for the GF-3 satellite are performed using ground control data from several regions. The experimental results show that the residual system errors of the GF-3 SAR satellite have been effectively eliminated, and the geometric positioning accuracy can be better than 3 m.

The loading and removal efficiency of 16 US EPA polycyclic aromatic hydrocarbons (PAHs) were examined in an inverted A2/O wastewater treatment plant (WWTP) located in an urban area in China. The total PAH concentrations were 554.3 to 723.2 ng/L in the influent and 189.6 to 262.7 ng/L in the effluent. The removal efficiencies of ∑PAHs in the dissolved phase ranged from 63 to 69%, with the highest observed in naphthalene (80% removal). Concentration and distribution of PAHs revealed that the higher molecular weight PAHs became more concentrated with treatment in both the dissolved phase and the dewatered sludge. The sharpest reduction was observed during the pretreatment and the biological phase. Noncarcinogenic risk, carcinogenic risk, and total health risk of PAHs found in the effluent and sewage sludge were also assessed. The effluent BaP toxic equivalent quantities (TEQBaP) were above, or far above, standards in countries. The potential toxicities of PAHs in sewage effluent were approximately 10 to 15 times higher than the acceptable risk level in China. The health risk associated with the sewage sludge also exceeded international recommended levels and was mainly contributed from seven carcinogenic PAHs. Given that WWTP effluent is a major PAH contributor to surface water bodies in China and better reduction efficiencies are achievable, the present study highlights the possibility of utilizing WWTPs for restoring water quality in riverine and coastal regions heavily impacted by PAHs contamination.

The atmospheric propagation delay of radar signals is a systematic error that occurs in the atmospheric environment, and is a key issue in the high-precision geometric calibration of spaceborne SAR. A multimode hybrid geometric calibration method for spaceborne SAR that considers the atmospheric propagation delay is proposed in this paper. Error sources that affect the accuracy of the geometric calibration were systematically analyzed. Based on correction of the atmospheric propagation delay, a geometric calibration model for spaceborne SAR was established. The high precision geometric calibration scheme for spaceborne SAR was explored by considering the pulse-width and bandwidth of the signal. A series of experiments were carried out based on high-resolution Yaogan 13 (YG-13) SAR satellite data and ground control data. The experimental results demonstrated that the proposed method is effective. The plane positioning accuracy of YG-13 in stripmap mode without control points is better than 3 m, and the accuracy of the sliding spotlight mode is better than 1.5 m.

This study investigated the prevalence and abundance of halogenated flame retardants (HFRs) in sludge samples from 5 sewage treatment plants in Guangzhou, China. Detection of 18 polybrominated diphenyl ethers (PBDEs), 9 alternative HFRs including Dechlorane Plus (DP), brominated alkylbenzenes, and polybrominated biphenyls, and 2 related degradation products was conducted. Decabromodiphenyl ether (BDE 209) and decabromodiphenyl ethane (DBDPE) were the dominant HFRs, with concentrations ranging from 200 to 2150 ng/g and 680–27,400 ng/g, respectively. The DBDPE detected was the highest level reported so far, exceeding those previously reported by 10–100 times. PBDEs were surpassed as the dominant HFRs in sewage sludge, with mean DBDPE/BDE 209 ratio exceeding 2 in all samples. The review of earlier surveys reveals that DBDPE level was surging while BDE 209 was declining. Annual emissions of BDE 209, DP, and DBDPE were estimated to be 227.9, 10.5, and 979.3 kg/yr, respectively. Although ecological risks assessment suggested low risks for the examined sludge, the key environmental properties and transformation pathways of alternative HFRs remain largely unknown. These findings prompt for further investigations on alternative HFR and sustainable management practices for HFR-laden biosolids. The HFR emission pattern revealed in this study is likely representative of other similarly industrialized regions in the post-PBDE era.

While inspecting animal feed for Salmonella contamination, we routinely observed bacterial colonies on selective agars that were similar in appearance to those formed by Salmonella. These were identified as Citrobacter freundii, Proteus mirabilis, and Serratia fonticola using biochemical and serological techniques. Because the presence of these bacterial species confounds identification of Salmonella, we refer to them as “interference bacteria.” Polyvalent antisera against these interference bacteria were prepared by immunizing rabbits with a mixture of all three organisms. To minimize or eliminate interference by these bacteria, the polyvalent antisera were introduced between the steps of selective enrichment and Salmonella-selective plating. The antisera raised against the interference bacteria, when combined with neonatal rabbit complement, exhibited specific bactericidal activity against C. freundii, P. mirabilis, and S. fonticola. The respective serum bactericidal assay titers were 29, 28, and 210. In selective broth, polyvalent antisera could also kill the target bacterial cells effectively. We tested 526 samples (186 white fishmeal, 97 red fishmeal, and 243 cattle bone powder) using the polyvalent antisera and found that the rates of contamination of each species of the three respective foods decreased by 58.8, 100, and 83%. Our data indicates that polyvalent sera against C. freundii, P. mirabilis, and S. fonticola can be used as inhibitors to increase the accuracy of Salmonella detection.

Nitrogen-footprint is a new method developed in recent years to measure the impact of reactive nitrogen produced by human activities on the environment. Livestock-product nitrogen-footprint, which is based on life cycle assessment theory (LCA) and nitrogen footprint, is used to define and measure the nitrogen emissions during various stages of livestock production process, and it aims to offer guidance for livestock producers to adopt rationale livestock breeding methods to reduce nitrogen footprint and further to promote the sustainable development of livestock industry. With different research scopes, livestock-product nitrogen- footprint can be used to measure the sustainable development of livestock industry from the following four dimensions: 1) to measure the ecological features of the livestock products; 2) to measure the ecological features of livestock breeding enterprises; 3) to measure the nitrogen footprint of the livestock breeding zone, 4) to exert the function of environmental protection education for consumers. It will shed some light on the further study of livestock nitrogen footprint.

There are many short-term road travel time forecasting studies based on time series, but indeed, road travel time not only relies on the historical travel time series, but also depends on the road and its adjacent sections history flow. However, few studies have considered that. This paper is based on the correlation of flow spatial distribution and the road travel time series, applying nearest neighbor and nonparametric regression method to build a forecasting model. In aspect of spatial nearest neighbor search, three different space distances are defined. In addition, two forecasting functions are introduced: one combines the forecasting value by mean weight and the other uses the reciprocal of nearest neighbors distance as combined weight. Three different distances are applied in nearest neighbor search, which apply to the two forecasting functions. For travel time series, the nearest neighbor and nonparametric regression are applied too. Then minimizing forecast error variance is utilized as an objective to establish the combination model. The empirical results show that the combination model can improve the forecast performance obviously. Besides, the experimental results of the evaluation for the computational complexity show that the proposed method can satisfy the real-time requirement.

The Gaofen-3 (GF-3) satellite is the first C-band multi-polarization synthetic aperture radar (SAR) with the ability of high-accuracy mapping in China. However, the Ground Control Points (GCPs) are essential to ensure the accuracy of mapping for GF-3 SAR imagery at present. In this paper, we analyze the error sources that affect the geometric processing and propose a new block adjustment method without GCPs for GF-3 SAR imagery. Firstly, the geometric calibration of GF-3 image is carried out. Secondly, the rational polynomial coefficient (RPC) model is directly generated after the geometric calibration parameters compensation of each image. Finally, we solve the orientation parameters of the GF-3 images through DEM assisted planar block adjustment and conduct ortho-rectification. With two different imaging modes of GF-3 satellite, which include the QPSI and FS2, we carry out the block adjustment without GCPs. Experimental results of testing areas including Wuhan city and Hubei province in China show that the geometric mosaic accuracy and the absolute positioning accuracy of the orthophoto are better than one pixel, which has laid a good foundation for the application of GF-3 image in global high-accuracy mapping.

Green agriculture can improve biodiversity, increase farmers’ income, reduce agricultural non-point source pollution, solve food safety issues, and will be an important way to promote sustainable development in China. At present, the green transformation of China’s agriculture has encountered a bottleneck in the development of a green agricultural product market. How to develop a green agricultural product market has become an issue worthy of in-depth study in the academia. Previous studies have already given persuasive explanations for the inability to form a green agricultural product market, but few have explored its development path from the angle of cooperation. By employing the method of a Transferable Utilities (TU) cooperative game, and based on theoretical analyses and hypothetical data, this thesis aims to prove the effectiveness of the multilateral coalition to develop the green agricultural product market in China. The results show the effectiveness of the developed model of the green agricultural product market in which producers, consumers, food safety inspection departments, and e-commerce platforms cooperate with each other. This model meets the objective needs of the times and that of the market economy. According to the marginal contribution value of participants in different coalition orders, this thesis finds 6 kinds of coalition orders. When producers and consumers of green agricultural products enter the coalition in the last place, the marginal contribution value is maximized, which reflects the importance of the supply side and demand side of green agricultural products. In other words, the development of the green agricultural product market is a dynamic process—determined by consumers and promoted by producers—in which both sides promote and restrict each other. Finally, this article presents two policy recommendations: at the national level, to clearly proposes a strategy to build a green agricultural product e-commerce platform in China and to launch a pilot application for the specialized e-commerce platform for green agricultural products in the Guizhou province.

Transcutaneous immunization is a novel strategy for genetic vaccine immunization to induce detectable antigen-special antibody in humor and mucosal. In this study, plasmid expressing hepatitis B surface antigen (pGFP-HBsAg) was encapsulated in liposome, then DNA- liposome complexes were glued on bare skin of mice ear in different dosage (50μg, 10μg and 1μg). As control, DNA- liposome complexes of pGFP-HBsAg and pGFP vector were inoculated intraperitoneally. The anti-HBsAg antibodies of serum were detected weekly by ELISA. It was found that the detectable antibodies of transcutaneous immunized mouse were elicited after four weeks, and reached a maximum at the sixth week. Even 1μg plasmid DNA in liposomes through immune skin can elicit the highest ELISA antibody titer (> 1:512) in test group, and corresponding percentage of positive response is up to 71% at sixth week, but higher amounts of plasmid DNA (50μg DNA per mice) on immune skin cannot induce higher antibody levels. The result showed that DNA- liposome complexes glued on bare skin appear to be a novel method for the administration of DNA vaccines.

The GaoFen-3 (GF-3) satellite is the first C-band multi-polarization synthetic aperture radar (SAR) imaging satellite with a resolution up to 1 m in China. It is also the only SAR satellite of the High-Resolution Earth Observation System designed for civilian use. There are 12 different imaging models to meet the needs of different industry users. However, to use SAR satellite images for related applications, they must possess high geometric accuracy. In order to verify the geometric accuracy achieved by the different modes of GF-3 images, we analyze the SAR geometric error source and perform geometric correction tests based on the RPC model with and without ground control points (GCPs) for five imaging modes. These include the spotlight (SL), ultra-fine strip (UFS), Fine Strip I (FSI), Full polarized Strip I (QPSI), and standard strip (SS) modes. Experimental results show that the check point residuals are large and consistent without GCPs, but the root mean square error of the independent checkpoints for the case of four corner control points is better than 1.5 pixels, achieving a similar level of geometric positioning accuracy to that of international satellites. We conclude that the GF-3 satellite can be used for high-accuracy geometric processing and related industry applications.

Twenty-one bacterial strains were isolated from imported cattle hide and rabbit wool using two types of media, nutrient broth, and nutrient broth with serum. The bacteria identified were Brevibacillus laterosporus, Leclercia adecarboxylata, Peptococcus niger, Bacillus circulans, Raoultella ornithinolytica, Bacillus subtilis, Bacillus cereus, Bacillus thermobacillus, Bacillus choshinensis, Bacillus sphaericus, Acinetobacter haemolyticus, Sphingomonas paucimobilis, Bacillus thuringiensis, Staphylococcus intermedius, Mycobacteria, Moraxella, Klebsiella pneumoniae, Ralstonia pickettii, Staphylococcus chromogenes, Comamonas testosteroni, and Cupriavidus pauculus. The 16s rDNA gene of each bacterium was amplified using the universal primers 27f and 1492r. The amplicons were digested with AvaI, BamHI, BgII, DraI, EcoRI, EcoRV, HindIII, HinfI, HpaI, PstI, SmaI, TaqI, XbaI, XmaI, AluI, XhoI, and PvuI individually. A specific fingerprint from the PCR-restriction fragment length polymorphism method based on 16s rDNA was obtained for each bacterium. The results showed that the method developed was useful not only for bacterial identification but also for the etiological investigation of pathogens in imported animal hair and wool.

The most important part of frame rate up-conversion (FRUC) is block matching. The geometric properties of the image were not taken into consideration in traditional block matching algorithm, so the matching result of motion estimation cannot reach the optimal. A novel FRUC algorithm based on Bandelet was proposed in this paper. The algorithm includes: Firstly, a soft threshold Bandelet transform of matching block was performed. The optimal matching block was determined through detection of direction similarity and Bandelet coefficient similarity; secondly, vector median filtering (VMF) and overlapped block motion compensation (OBMC) were carried out by adopting motion vector to realize interpolated frame algorithm. Experimental results show that the FRUC algorithm based on Bandelet can further promote the quality of FRUC.

Different molecular detection methods require diverse molecular platforms, but there is no uniform standard for people to reference in the detection of Trichinella. In this study, real-time PCR, loop-mediated isothermal amplification (LAMP), and conventional PCR were developed for the detection of Trichinella by targeting mitochondrial large subunit ribosomal DNA (mt-lsrDNA). We compared the performance of the three newly developed assays. The results revealed that the detection limits of the real-time PCR, LAMP, and conventional PCR assays were 10 and 100 fg/μL and 1 pg/μL of Trichinella spiralis genomic DNA, respectively. The assays were used in the detection of Trichinella in the field. A total of 192 samples were obtained from pigs: 75 samples from free range farming and 117 from intensive feeding factory. The infection rate was 8/192 (4.2 %), 7/192 (3.6 %), and 1/192 (1.0 %) through the real-time PCR, LAMP, and conventional PCR assays, respectively. These data indicate that Taqman real-time PCR was a rapid, specific, and sensitive tool as a preferred option for investigation of valuable samples, but that LAMP assay was closed tube, highly sensitive, cost-effective, rapid, easy-to-perform, and was the optimal choice for detection of Trichinella in the field. The results of a model of experimental infection in mice indicated that spleen can be used as sampling site for the detection of early T. spiralis infection. However, the diaphragm and myocardium were the most suitable sampling sites for the detection of T. spiralis.

To eliminate the interference caused by Pseudomonas aeruginosa in the isolation of Salmonella, a rabbit polyclonal antibody against P. aeruginosa was prepared by inoculating four New Zealand rabbits with the pathogen. The antiserum was purified using saturated ammonium sulfate and added into Rappaport-Vassiliadis medium with soya (RVS) broth and Muller-Kauffmann tetrathionate novobiocin broth (MKTTn broth) to evaluate whether it could inhibit the growth of P. aeruginosa. Observations by scanning electron microscopy demonstrated that P. aeruginosa was attacked and destroyed by the antibody when incubated for 10 min at 37°C. The activity of the antibody was also effective against 11 other strains of P. aeruginosa. Twenty-six strains of Salmonella were mixed with P. aeruginosa in RVS and MKTTn broth at 37°C for 12 h, respectively, and the cultures were plated on Salmonella chromogenic medium (SCM; Oxoid, Basingstoke, UK). Only Salmonella grew on SCM; five colonies were randomly selected for identification by VITEK 2 (bioMérieux, Lyon, France). Additionally, when mixed with two strains of Enterobacter cloacae (ATCC 700323 and YG001), the prepared antibody did not affect the growth of E. cloacae. The results demonstrated that the microbicidal activity of the antibody did not affect the tested Salmonella sp. or E. cloacae strains. Therefore, the antibody generated could be used to increase the accuracy of Salmonella isolation.

The aim of the present work was to develop a quantitative reverse-transcription polymerase chain reaction (qRT-PCR) assay using a TaqMan probe to detect and quantify hirame rhabdovirus (HRV). The results demonstrated that the assay had a detection limit of 100 copies of RNA per reaction and a log-linear range up to 10(8) copies of HRV RNA. Regression analysis demonstrated a significant correlation with an R(2) value of 0.9963 and a slope of -3.18 between the mean C(t) values and HRV cRNA. This assay was 100 times more sensitive than the conventional one-step RT-PCR assay. The qRT-PCR assay was found to be highly reproducible with intra- and inter-assay coefficients of variation of 0.37-1.72% and 1.37-3.79%, respectively. The primers and TaqMan probe were specific for HRV and did not react with either the spring viraemia of carp virus (SVCV), infectious pancreatic necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV), marine birnavirus (MABV), viral hemorrhagic septicemia virus (VHSV), or viral nervous necrosis virus (VNNV). This assay was evaluated using 40 fish samples, indicating that such method offers considerable advantages over the classical virus isolation method currently used for HRV surveillance. In conclusion, the developed qRT-PCR assay was a reliable, specific and sensitive tool for the quantitative diagnosis of HRV in fish samples.

Fish, as one of the most common causes of IgE-mediated food hypersensitivity, has recently received increasing attention from the food industry and legislative and regulatory agencies. A real-time polymerase chain reaction assay based on TaqMan-MGB probe technology was developed for the detection of parvalbumin, a major fish allergen gene. The assay had a sensitivity up to 5 pg purified fish DNA and had no cross-reaction with other species, such as cattle, sheep, swine, chicken, shrimp, lobster, crab, squid, clam, rice, soybean, maize, and potato. The coefficient of variation for both intra- and interexperimental variability demonstrated high reproducibility and accuracy. The assay proved to be a potential tool for the detection and label management of fish allergens in food.