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In recent decades, the frequency and intensity of harmful algal blooms (HABs), as well as a profusion of toxic phytoplankton species, have significantly increased in coastal regions of China. Researchers attribute this to environmental changes such as rising atmospheric CO2 levels. Such addition of carbon into the ocean ecosystem can lead to increased growth, enhanced metabolism, and altered toxicity of toxic phytoplankton communities resulting in serious human health concerns. In this study, the effects of elevated partial pressure of CO2 (pCO2) on the growth and toxicity of a strain of Alexandrium tamarense (ATDH) widespread in the East and South China Seas were investigated. Results of these studies showed a higher specific growth rate (0.31±0.05day-1) when exposed to 1000μatm CO2, (experimental), with a corresponding density of (2.02±0.19)×107cellsL-1, that was significantly larger than cells under 395μatm CO2(control). These data also revealed that elevated pCO2 primarily affected the photosynthetic properties of cells in the exponential growth phase. Interestingly, measurement of the total toxin content per cell was reduced by half under elevated CO2 conditions. The following individual toxins were measured in this study: C1, C2, GTX1, GTX2, GTX3, GTX4, GTX5, STX, dcGTX2, dcGTX3, and dcSTX. Cells grown in 1000μatm CO2 showed an overall decrease in the cellular concentrations of C1, C2, GTX2, GTX3, GTX5, STX, dcGTX2, dcGTX3, and dcSTX, but an increase in GTX1 and GTX4. Total cellular toxicity per cell was measured revealing an increase of nearly 60% toxicity in the presence of elevated CO2 compared to controls. This unusual result was attributed to a significant increase in the cellular concentrations of the more toxic derivatives, GTX1 and GTX4.Taken together; these findings indicate that the A. tamarense strain ATDH isolated from the East China Sea significantly increased in growth and cellular toxicity under elevated pCO2 levels. These data may provide vital information regarding future HABs and the corresponding harmful effects as a result of increasing atmospheric CO2.

Vibrio parahaemolyticus is a kind of food-borne pathogenic bacterium, which can seriously infect food, especially seafood causing gastroenteritis and other disease. We studied the global proteome responses of V. parahaemolyticus under cold stress by nano-liquid chromatography-tandem mass spectrometry to improve the present understanding of V. parahaemolyticus proteomics events under cold stress. A total of 1151 proteins were identified and 101 proteins were differentially expressed, of which 69 were significantly up-regulated and 32 were downregulated. Functional categorization of these proteins revealed distinct differences between cold-stressed and control cells. These proteins were grouped into 21 functional categories by the clusters of orthologous groups (COG) analysis. The most of up-regulated proteins were functionally categorized as nucleotide transport and metabolism, transcription, function unknown, and defense mechanisms. These up-regulated proteins play an important role under cold stress.

A multi-residue procedure with liquid chromatography-tandem mass spectrometry analysis was developed to simultaneously detect the presence of 4 diarrheic shellfish poisoning toxins, 14 sulfonamides, chloramphenicol and thiamphenicol in shellfish tissues. Samples were first purified by a modified rapid procedure and separated on a ZOBRAX Eclipse Plus C18 column and quantified using electron spray ionization-mass spectrometry-mass spectrometry. The results showed that the limits of quantification (LOQ) of SPX1, OA-C, GYM, and PTX-2 were 1.0, 2, 0.5 and 2 μg/kg, respectively. The LOQ for the antibiotics was below acceptable limits established by most countries. Compared to the external standard method, this matrix matched the calibration method effectively, overcame the matrix effects and gave better quantitative results. Recoveries of spiked compounds ranged between 67.6% and 109.8%, with relative standard deviations below 15% for most target analytes. Only sulfathiazole had a %RSD of 18.6% at the lowest spiked concentration. The proposed method was accurate, rapid and reliable.https://doi.org/10.21423/jrs-v05n01p009 (DOI assigned 4/17/2019)

Cascaded H-bridge (CHB) inverters used in high-voltage variable frequency drives are usually composed of several power cells. Each power cell consists of a three-phase uncontrolled rectifier as its input stage and a single-phase inverter as its output stage. It is difficult to obtain the analytical expression of the dc-link capacitor current because of both the nonlinear characteristics of the uncontrolled rectifier and the low-frequency harmonics caused by the single-phase inverter. On the other hand, exact analysis of the capacitor current is important for dc-link capacitor selection. This paper proposes an analytical method for calculating the capacitor current. Considering both the input current of the inverter and the output current of the rectifier, an approximately linear equivalent circuit model is established. According to the proposed equivalent circuit model, the analytical expressions of the harmonic spectrum and the root mean square value of the capacitor current are derived. The capacitor ripple voltage is further estimated. Applicable conditions of the proposed method are also presented. This method may be useful when designing dc-link capacitors of power cells in CHB inverters. Simulation and experimental results show that the proposed method can achieve good accuracy in the continuous conduction mode.

Detecting avian influenza virus (AIV) and Newcastle disease virus (NDV) at low concentrations from tracheal and cloacal swabs of avian influenza- and Newcastle disease-infected poultry was carried out using a highly sensitive immunological-polymerase chain reaction (immuno-PCR) method. Magnetic gold particles were pre-coated with a capture antibody, either a monoclonal anti-AIV/H5 or monoclonal anti-NDV/F and viruses serially diluted ten-fold from 102 to 10−5 EID50/ml. A biotinylated detection antibody bound to the viral antigen was then linked via a streptavidin bridge to biotinylated reporter DNA. After extensive washing, reporter DNA was released by denaturation, transferred to PCR tubes, amplified, electrophoresed and visualized. An optimized immuno-PCR method was able to detect as little as 10−4 EID50/ml AIV and NDV. To further evaluate the specificity and the clinical application of this IPCR assay for AIV H5N1 and NDV, the tracheal swab specimens, taken from chickens which were infected with H5N1/AIV, H9N2/AIV, H7N2/AIV, NDV, IBDV, IBV/H120, were detected by IPCR. Our data demonstrated that this monoclonal antibody-based immuno-PCR method provides a platform capable of rapid screening of clinical samples for trace levels of AIV H5 and NDV in one step.

Constructed wetland (CW) is a low cost and easy operation process for wastewater treatment, while filler clogging is one of the disadvantages for this technology. Using activated carbon as the filler, a regular CW and an integration of constructed wetland with microbial fuel cell (CW-MFC) were constructed. After continuous operation of four months, specific surface area and pore size distribution of the activated carbon were analyzed. The specific surface area of the fresh activated carbon was 133.8 m2/g, while the filler in the CW and CW-MFC systems had specific surface area of 38.1 m2/g and 58.2 m2/g. The surface decrease of the CW-MFC filler was 21% lower than that of CW filler. Comparing with the regular CW, the filler in the CW-MFC was more clogging resistant. The alleviation of the filler clogging in CW-MFC may be caused by the micro-electric field as the function of MFC. CW-MFC is a promising microbial electrochemical technology for wastewater treatment and filler clogging resistant, there are some detailed issues deserve to be further researched.

Zooplankton is an important link between phytoplankton and higher consumers in the marine food chain. To investigate the harmful effects of the toxic dinoflagellate Alexandrium species on zooplankton, 4 strains of Alexandrium spp., isolated from the Chinese coast, were used to test the species' effects on the survival and feeding rates of the brine shrimp, Artemia salina. The experiment was designed to assess the response of A. salina in each stage of its life cycle: nauplii, metanauplii, and adult. Each experiment was conducted in a 500 ml treatment that was added. The toxic treatments consisted of single strains of A. minutum, A. catanella, and A. tamarense (Nanhai and Donghai strain), while non-toxic species (dinoflagellate Prorocentrum donghaiense and diatom Chaetoceros minutissimus) were used as control treatments. An additional phytoplankton treatment consisted of, a mixture of A. tamarense (Nanhai strain) and P. donghaiense. Alexandrium spp. species were found to have lethal effects on the brine shrimp at a density of 2000 cells/ml. All the brine shrimps died within 24-168 hours of inoculation with the 4 treatments each containing single toxic Alexandrium species. During the feeding experiment, toxic Alexandrium spp. caused a reduction in the feeding rates in all the three stages of the life cycle of A. salina, whereas this response was not obvious in the treatment involving the nontoxic species P. donghaiense. The body surface of the brine shrimp that were fed on Alexandrium species was consistently covered by a sticky floc. Mortality of A. salina was observed to increase with the occurrence of the floc. The toxicity of the paralytic shellfish poisons (PSP) produced by the Alexandrium species was not significantly correlated with the survival or the feeding rate of the brine shrimp. When A. tamarense was mixed with P. donghaiense, the lethal effect of A. tamarense decreased, as shown by an increase in the survival and the feeding rates of the brine shrimp. A. salina metanauplii were found at the life stage most sensitive to the toxic algae and hunger. In summary, toxic Alexandrium spp. were found to have lethal effects on A. salina and to restrain the brine shrimp's feeding rate. Nontoxic Prorocentrum mitigated the toxicity of Alexandrium to a certain extent. The results also imply that the sticky material on the surface of the body of the brine shrimp may have been an important lethal factor rather than the PSP toxins.

Simple and low cost sensor for the determination of bisphenol A (BPA) based on graphene-1-butyl-3-methylimidazolium hexafluorophosphate (BmimPF6) modified glassy carbon electrode was developed. It was an irreversible oxidation process of BPA on the modified electrode. Experimental conditions such as modified volume, pH, scan rate and accumulation time have been optimized. The modified electrode provided a considerable enhancement of BPA oxidation. The electrochemical response of BPA on this modified electrode was better than those on the graphene modified electrode and bare electrode. The detection limit of BPA was 8nM (S/N=3), the linear range was from 20nM to 2µM. The modified electrode has been employed for determination of milk and soda spiked BPA successfully.

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 accuracy, repeatability, and reproducibility characteristics of a method using immunoaffinity column (IAC) cleanup with postcolumn derivatization and LC with a fluorescence detector (FLD) for determination of aflatoxins (AFs; sum of AFs B1, B2, G1, and G2) in olive oil, peanut oil, and sesame oil have been established in a collaborative study involving 15 laboratories from six countries. Blind duplicate samples of blank, spiked at levels ranging from 0.25 to 20.0 μg/kg for AF, were analyzed. A naturally contaminated peanut oil sample was also included. Test samples were extracted with methanol–water (55 + 45, v/v). After shaking and centrifuging, the lower layer was filtered, diluted with water, and filtered through glass microfiber filter paper. The filtrate was then passed through an IAC, and the toxins were eluted with methanol. The toxins were then subjected to RPLC-FLD analysis after postcolumn derivatization. Average recoveries of AFs from olive oil, peanut oil, and sesame oil ranged from 84 to 92% (at spiking levels ranging from 2.0 to 20.0 μg/kg); of AFB1 from 86 to 93% (at spiking levels ranging from 1.0 to 10.0 μg/kg); of AFB2 from 89 to 95% (at spiking levels ranging from 0.25 to 2.5 μg/kg); of AFG1 from 85 to 97% (at spiking levels ranging from 0.5 to 5.0 μg/kg); and of AFG2 from 76 to 85% (at spiking levels ranging from 0.25 to 2.5 μg/kg). RSDs for within-laboratory repeatability (RSDr) ranged from 3.4 to 10.2% for AF, from 3.5 to 10.9% for AFB1, from 3.2 to 9.5% for AFB2, from 6.5 to 14.9% for AFG1, and from 4.8 to 14.2% for AFG2. RSDs for between-laboratory reproducibility (RSDR) ranged from 6.1 to 14.5% for AF, from 7.5 to 15.4% for AFB1, from 7.1 to 14.6% for AFB2, from 10.8 to 18.1% for AFG1, and from 7.6 to 23.7% for AFG2. Horwitz ratio values were ≤2 for the analytes in the three matrixes.

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.

Textile electrode is flexible, folding, washable and biocompatible with skin. With these advantages, the textile electrodes should be an ideal alternative for electromyogram (EMG) recordings in clinical applications. In this study, a textile electrode system was used for EMG signal recordings and its usability and performance in classifying different arm and hand movements were evaluated through real-time pattern recognition control of virtual myoelectric arm. Our results showed that the offline average classification accuracy over seven able-bodied subjects for all ten movements was 91.81%, compared to 96.54% for six basic movements. The real-time performance in operating a virtual arm was quantified with three measures: selection time, completion time, and completion time. Averagely, the seven subjects achieved a motion selection time of approximately 0.36 s, and a completion time of less than 2 s, and a completion rate of around 82%. These pilot results suggest that the textile electrodes could work well in real-time control of multifunctional myoelectric prosthesis.