Fate of sulfamethoxazole and potential formation of haloacetic acids during chlorine disinfection process in aquaculture water.

There exist two common processes in fishery culture, i.e. antibiotic addition to reduce disease in fishery, and chlorination disinfection to inhibit infectious pathogenic microorganisms. However, antibiotic residues might play important reverse side roles for both aquaculture water pollution and potential formation of chlorination side products. Herein, the transformation behaviour, intermediates analyses and conversion pathway of antibiotic sulfamethoxazole (SMX), and potential generation of halogenated acetic acids (HAAs) in the process of chlorination in fishery water were examined, and the results revealed that the decomposing of SMX satisfied a pseudo first-order kinetic equation. Both the addition of available chlorine and high temperature had affirmative influences on the decontamination of SMX and production of HAAs, and the near-neutral pHs promoted the removal of SMX and generation of HAAs. Br- was favorable for the removal of SMX and yields of brominated acetic acids (Br-AAs). Based on the identified intermediate products, the transformation path of SMX in chlorination process was propounded, to wit, the C-S and S-N bonds in the SMX molecules were firstly cracked, and the primeval intermediate groups are then transformed to form chloroanilines, chlorophenols, etc., and subsequently, chlorophenols were chlorinated and ring-opened to generate toxic HAAs. This study might be meaningful to evaluate the effective removal of sulfonamide antibiotic residues and the potential generation of halogenated DBPs (H-DBPs) when chlorinated in aquaculture water.

Determination of Methylene Blue and Its Metabolite Residues in Aquatic Products by High-Performance Liquid Chromatography-Tandem Mass Spectrometry.

A sensitive and reliable method was developed to determine methylene blue (MB) and its metabolite residues, including azure A (AZA), azure B (AZB), and azure C (AZC) in aquatic products by HPLC-MS/MS. The samples were extracted by acetonitrile and cleaned up by alumina-neutral (ALN) cartridges. The analytes were separated on a Sunfire C18 column (150 mm × 2.1 mm, 5 µm). The method was validated according to the European criteria of Commission Decision 2002/657/CE. Good linearity between 1-500 µg/L was obtained with correlation coefficients (R2) greater than 0.99. The limit of quantification (LOQ) was 1.0 µg/kg. The average recoveries at three levels of each compound (1, 5, and 10 µg/kg) were demonstrated to be in the range of 71.8-97.5%, with relative standard deviations (RSDs) from 1.05% to 8.63%. This method was suitable for the detection of methylene blue and its metabolite residues in aquatic products.

"Switch-Off-On" Detection of Fe3+ and F- Ions Based on Fluorescence Silicon Nanoparticles and Their Application to Food Samples.

An approach to the detection of F- ions in food samples was developed based on a "switch-off-on" fluorescence probe of silicon nanoparticles (SiNPs). The fluorescence of the synthetic SiNPs was gradually quenched in the presence of Fe3+ ion and slightly recovered with the addition of F- ion owing to the formation of a stable and colorless ferric fluoride. The fluorescence recovery exhibited a good linear relationship (R2 = 0.9992) as the concentration of F- ion increased from 0 to 100 µmol·L-1. The detection limit of the established method of F- ion was 0.05 µmol·L-1. The recovery experiments confirmed the accuracy and reliability of the proposed method. The ultraviolet-visible spectra, fluorescence decays, and zeta potentials evidenced the fluorescence quenching mechanism involving the electron transfer between the SiNPs and Fe3+ ion, while the fluorescence recovery resulted from the formation of ferric fluoride. Finally, SiNPs were successfully applied to detect F- ions in tap water, Antarctic krill, and Antarctic krill powder.

Simultaneous Determination of Tetrodotoxin in the Fresh and Heat-Processed Aquatic Products by High-Performance Liquid Chromatography-Tandem Mass Spectrometry.

Tetrodotoxin (TTX) was simultaneously detected in the fresh and heat-processed aquatic products by high-performance liquid chromatography-tandem mass spectrometry method. The detection conditions were investigated, including the chromatography column and mobile phase. Based on the optimized parameters, a sensitive determination method of TTX was established. The proposed method featured the merits of a good linear relationship between signal and TTX concentration (R2 = 0.9998), a wide detection matrix-based range of 0.2-100 ng/g, and a low detection limit of 0.2 ng/g, etc. The spiked assays evidenced its accuracy and reliability with recoveries of 90.5-107.2%. Finally, the developed method was simultaneously successfully applied in the determination of TTX in various fresh and heat-processed aquatic products.

Simulation experiment on OH-PCB being ingested through daily diet: Accumulation, transformation and distribution of hydroxylated-2, 2', 4, 5, 5'-pentachlorobiphenyl (OH-PCB101) in mice.

Animals exposure to polychlorinated biphenyls (PCBs) may result in retention of hydroxylated PCBs (OH-PCBs). OH-PCBs can be accumulated in animals, including humans, through the transmission of food chain. However, there are few studies on the accumulation and metabolism of OH-PCBs exposed to the body through daily diet. Therefore, this study was conducted to investigate the fate of OH-PCBs after being ingested through dietary intake. By adding 3-OH-PCB101 and 4-OH-PCB101 to the edible tissue of crucian carp, which were used as raw materials to prepare mouse feed, with an exposure concentration of 2.5 µg/kg ww. The exposure experiment lasted for a total of 80 days. The blood, feces and 11 tissues of mice at different times were analyzed qualitatively and quantitatively. It was found that major OH-PCB101 were accumulated in intestine or excreted with feces. A small part was accumulated in heart, lung and spleen. For the first time that the conversion from OH-PCB101 to PCB101 in mice was discovered, which shows from another perspective that persistent organic pollutants are difficult to be completely degraded in the environment. 4-MeO-PCB101, 3-MeSO2-PCB101, and 4-MeSO2-PCB101 were also found in various tissues. The results of this study show that after OH-PCBs accumulated in animals re-enter the organism through the food chain, they can be metabolized again and may be reversely transformed into the parent compounds. The present research shed new light on simulating the metabolic transformation process of OH-PCBs exposed to mammals through ingestion of fish. Available data show that second-generation persistent organic pollutants in the environment still need to be continuously concerned.

Identification of 2,2',4,5,5'-Pentachlorobiphenyl (PCB101) metabolites and their transmission characteristics in silver crucian carp (Carassius auratus gibelio).

Polychlorinated biphenyls (PCBs) have been attracting global concern due to their persistence and toxicity. However, the study on the metabolites of PCBs in freshwater fish is limited. In this study, the metabolites of 2,2',4,5,5'-Pentachlorobiphenyl (PCB101) in silver crucian carp (Carassius auratus gibelio) were identified for the first time. After intraperitoneal injection of PCB101 (2 mg/kg), the results showed that it could be metabolized to at least three types of metabolites, including hydroxylated (OH-), methoxylated (MeO-) and methyl sulfonated (MeSO2-) PCB101. The OH- metabolites identified in most tissues were 3-OH-PCB101and 4-OH-PCB101, such as liver, gallbladder, blood and muscle. MeSO2- metabolites identified in gallbladder, blood and brain were 3-MeSO2-PCB101 and 4-MeSO2-PCB101. Meanwhile, the MeO- metabolite identified in liver, gallbladder, blood and spleen of silver crucian carp was 4-MeO-PCB101. The investigation of the types and structures of PCB101 and its metabolites, as well as the tissue distribution and accumulation characteristics in silver crucian carp are beneficial to understand the transformation and metabolic mechanisms of PCBs in aquatic organisms. It is of great significance to identify potential pollution hazards of precursor compounds and their metabolites on aquatic products and ensure the quality and safety of aquatic products.

Dehydrogenation of methylcyclohexane over Pt-based catalysts supported on functional granular activated carbon.

Herein, we developed the dehydrogenation of methylcyclohexane over Pt-based catalysts supported on functional granular activated carbon. Sulphuric acid, hydrogen peroxide, nitric acid and aminopropyl triethoxy silane were adopted to modify the granular activated carbon. The structural characterizations suggested that the carbon materials had a large surface area, abundant pore structure, and a high number of oxygen-containing functional groups, which influenced the Pt-based catalysts on the particle size, dispersion and dehydrogenation activity. The hydrogen temperature-programmed reduction technique was utilized to investigate the interaction between the active component Pt and the various functionalized granular activated carbon materials. The CO pulse technique revealed the particle sizes and dispersion of the as-prepared Pt-based catalysts. Finally, the Pt-based catalysts were successfully applied to study their catalytic activity in the dehydrogenation reaction of methylcyclohexane. The results showed that the Pt-based catalyst over granular activated carbon functionalized with sulphuric acid groups had a higher conversion of methylcyclohexane (63%) and a larger hydrogen evolution rate (741.1 mmol gPt -1 min-1) than the other resulting Pt-based catalysts at 300 °C.

Determination of 6 biogenic amines in food using high-performance liquid chromatography-tandem mass spectrometry without derivatization.

A rapid and simple method for the determination of 6 biogenic amines (BAs) in food was established on HPLC-MS /MS without derivatization. Samples were extracted with 5% perchloric acid and cleaned with n-hexane for lipid removal. The analytes were separated on Waters XBridge® HILIC (150 mm × 2.1 mm, 3.5 µm) and analyzed with multiple-reaction monitoring (MRM) mode after positive electrospray ionization on HPLC-MS/MS. Good linearity with high correlation coefficient was obtained between 10-1000 µg/L for cadaverine (CAD), putrescine (PUT), tyramine (TYR) and 2-phenylethylamine (2-PHE) and between 1-100 µg/L for histamine (HIS) and tryptamine (TRY), with the detection limits of the method ranging from 0.1 mg/kg for HIS and TRY, and 1.0 mg/kg for CAD, PUT, TYR and 2-PHE, which are under the residue limit of Chinese regulation. Spiking experiments demonstrated good recoveries between 70.2-114.6%, with relative standard deviations (RSDs) between 0.44-13.01%. This method was validated for BAs determination in liquor, fermented meat products, vegetable products, soybean products, dairy products, seafood and its derived products. These results promise high feasibility for BAs monitoring in various food with easy-to-operate and fast sample preparation process, stable analysis on HPLC-MS/MS without derivatization.

Determination of 8 biogenic amines in aquatic products and their derived products by high-performance liquid chromatography-tandem mass spectrometry without derivatization.

A method for the determination of 8 biogenic amines in aquatic products and their derived products was established by HPLC-MS/MS without derivatization. The samples were extracted by 5% perchloric acid solution. N-hexane was used to clean the extract. The analytes were separated by a column of ACQUITY UPLC HSS T3 (100 mm × 2.1 mm, 1.8 µm), and gradient eluted with a mixed solution of (0.5% formic acid) and acetonitrile. Good linearity was obtained with correlation coefficients (R2) >0.99. This method achieved higher sensitivity (from 0.1 mg/kg for tyramine, 2-phenylethylamine and tryptamine to 1.0 mg/kg for spermidine, spermine, cadaverin, histamine and putrescine). The average recoveries were demonstrated in the range of 70.9%-113.1%, with relative standard deviations (RSDs) from 0.33% to 10.81%. This method was suitable for the detection of BAs in aquatic products and their products.

An Effective and Efficient Sample Preparation Method for 2-Methyl-Isoborneol and Geosmin in Fish and Their Analysis by Gas Chromatography-Mass Spectrometry.

The intensive aquaculture strategy and recirculating aquaculture system often lead to the production of off-flavor compounds such as 2-methyl-isoborneol (2-MIB) and Geosmin (GSM). The regular purge and trap extraction followed by analysis with gas chromatography-mass spectrometry (GC-MS) usually involve a complicated assembly of facilities, more working space, long sample preparation time, and headspace solid-phase microextraction (SPME). In this work, a method with easier sample preparation, fewer and simplified facilities, and without SPME on GC-MS analysis is developed for the determination of 2-MIB and GSM in fish samples. Unlike previous methods, solvent extract from samples, QuEChERS-based cleanup, and solid-phase extraction for concentration are applied. The LOD (S/N > 3) and LOQ (S/N > 10) of this method were validated at 0.6 µg/kg and 1.0 µg/kg for both 2-MIB and GSM, which are under the sensory limit (1 µg/kg). Application of this method for incurred fish samples demonstrated acceptable analytical performance. This method is suitable for large-scale determination of 2-MIB and GSM in fish samples, owing to the use of simple facility and easy-to-operate procedure, rapid sample preparation, and shorter time for GC-MS analysis without SPME.

Characterization of the complete mitochondrial DNA sequence of the Lagocephalus guentheri (Tetraodontidae, Tetraodontiformes).

The complete mitochondrial genome of Lagocephalus guentheri was reported in the present study, which was 16,461 bp in length. It consists of 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes and a non-coding control region. The overall base composition of the genome is 27.54% for A, 24.80% for T, 31.23% for C and 16.43% for G. The phylogenetic tree, which is based on 12 protein-coding gene sequences, suggested that L. guentheri was closest to L. spadiceus. This study could give impetus to studies focused on population structure and molecular evolution of L. guentheri.

Characterization of the complete mitochondrial DNA sequence of the Lagocephalus gloveri (Tetraodontidae, Tetraodontiformes).

The complete mitochondrial genome of Lagocephalus gloveri is reported in the present study, which is 16,446 bp in length. It consists of 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes and a non-coding control region. The overall base composition of the genome is 27.58% for A, 25.07% for T, 30.83% for C and 16.52% for G. The phylogenetic tree, which is based on 12 protein coding gene sequences, suggested that L. gloveri was closest to L. lagocephalus. This study could give impetus to studies focused on population structure and molecular evolution of L. gloveri.

Detection of Total and Pathogenic Vibrio parahaemolyticus in Shellfish Growing along the South Yellow Sea and the East China Sea.

This study was conducted to monitor the densities of total and pathogenic Vibrio parahaemolyticus in 300 samples of nine shellfish species harvested from the coasts of the South Yellow Sea and the East China Sea (N 23° to 34°, E 116° to 124°), People's Republic of China, between May and October 2015. Total V. parahaemolyticus densities were measured, and V. parahaemolyticus isolates were biochemically identified with probes for the thermostable direct hemolysin gene (tdh) and the thermostable direct hemolysin-related hemolysin gene (trh). We found that 202 of the 300 samples were positive for V. parahaemolyticus from all the sites: 58 of the 100 samples from the Fujian province, 71 of the 100 samples from the Zhejiang province, and 73 of the 100 samples from the Jiangsu province. In most (170) of the 300 samples, V. parahaemolyticus densities were 0.3 to 10 most probable number (MPN)/g; five lots exceeded 110 MPN/g, and two lots were estimated at 110 MPN/g. Among the 202 V. parahaemolyticus strains, only one was trh positive. Densities of V. parahaemolyticus in these shellfish were temperature dependent, with highest densities in June and July. Among the nine mollusk species, V. parahaemolyticus was most abundant in the agemaki clam (Sinonovacula constricta). The highest and lowest V. parahaemolyticus prevalences were found in oriental cyclina (Cyclina sinensis, 93.8%) and mussels (Mytilus edulis, 28.1%), respectively. Overall, although V. parahaemolyticus is widely distributed in marine environments, the density of V. parahaemolyticus was low and the prevalence of the main virulence factor was very low in shellfish along the coasts of the South Yellow Sea and East China Sea, which is important from a public health perspective. Data presented here will be useful for correlational research and can be utilized for developing risk management plans that establish food safety guidelines for V. parahaemolyticus in Chinese shellfish.

[Identification of Vibrio campbellii isolated from cultured pacific oyster].

Four bacterial strains were isolated from cultured pacific oyster (Crassostrea gigia) in September 2003 at coast of Fujian province. Their morphological, physiological and biochemical characteristics and 16S rRNA sequence were analyzed. And the relationship between reproduction of the bacterium and NaCl concentrations, pH and temperature were also determined. The results showed that four strains were gram-negative rods with a single polar flagellum, glucose fermented without gas production, oxidase positive, required sodium ions for growth, no pigment, non-luminescence; They grew well on TCBS-plate as green colonies and were sensitive to the vibriostratic agent O/129. Therefore, it was confirmed that the strains belonged to the genus of Vibrio. The sequence analysis of 16S rRNA gene of SXS1 isolation and comparison with that of other related vibrios- showed that SXS1 was very close to Vibrio campbellii. The similarities was 99%. The distribution of V. campbellii in environment and its relationship to the diseases of aquaculture animals were discussed.

Chitosan-based adsorption and freeze deproteinization: Improved extraction and purification of synthetic colorants from protein-rich food samples.

A freeze method for deproteinization coupling with the chitosan purification process was developed for the determination of 8 synthetic food colorants in protein-rich samples. The solvents for extraction and different methods for deproteinization were examined and selected. Chitosan was employed for the purification after deproteinization, and further compared with the traditional polyamine purification method. Determination of the purified extract was conducted through the separation using high performance liquid chromatography and detection by multi-wavelength mode. Under the optimum conditions, the method showed good linearity between 0.6 and 10mg/kg, for the 8 synthetic colorants, and the limit of detection was between 0.1 and 0.4 mg/kg as was defined when the ratio of signal to noise was three. The recoveries of the spiked samples were found to be between 83% and 91%. The intra-day precision and inter-day precision was estimated to be 3-10% and 6-12%, respectively. The developed method could be applied to deproteinization and clean-up for pretreatment of protein-rich samples.

[Determination of histamine in canned fish by high performance liquid chromatography with pre-column derivatization].

A pre-column derivatization-high performance liquid chromatographic (HPLC) method has been developed for the determination of histamine in canned fish. The homogenated samples were ultrasonically extracted with perchloric acid aqueous solution, derivatized with dansyl chloride and diluted with acetonitrile to a desired volume. The samples were determined by HPLC with ultraviolet detector and quantified by external standard method. Adopting a C18 column with 1.8 microm stationary phase particles, the analysis time for each sample was smaller than 5 min with the flow rate of 0.3 mL/min. It can decrease the consumption of the mobile phase and save the cost. The linear range was 0.08-8.00 mg/L for histamine. The correlation coefficient was 0.999 98. The average recoveries of histamine at different concentration levels in spiked samples were greater than 96% and the relative standard deviations (RSDs) were smaller than 2.5%. The quantitation limit was 5.00 mg/kg for histamine in canned fish by HPLC. The results indicated that this HPLC method is fast, sensitive, reproducible and practical for the routine analysis of histamine in canned fish.

Effect of temperature on uptake and survival of Vibrio parahaemolyticus in oysters (Crassostrea plicatula).

This study investigated accumulation of Vibrio parahaemolyticus in Zhe oyster (Crassostrea plicatula) from culture water and effectiveness of frozen and chilled storage on reducing V. parahaemolyticus in oysters. Freshly harvested oysters were placed in artificial seawater containing V. parahaemolyticus (10(4)CFU/mL) at 16, 20, 26, and 32 degrees C for 96 h. Contaminated oysters were stored at chilled temperatures (0, 5, and 15 degrees C) and frozen at -18 and -30 degrees C and changes of V. parahaemolyticus populations in oysters were determined using the most probable number (MPN) method. Accumulations of V. parahaemolyticus in C. plicatula reached the peaks at 6.66 (32 degrees C), 5.72 (26 degrees C), 5.04 (20 degrees C), 4.72 (16 degrees C) log MPN/g after 32 h in contaminated artificial seawater. Holding contaminated Zhe oysters at 5 and 0 degrees C reduced V. parahaemolyticus populations in both shell stock and shucked oysters. Populations of V. parahaemolyticus in shell stock and shucked oysters declined by 1.42 and 2.0 log MPN/g, respectively, after 96 h of storage at 5 degrees C and by 2.11 and 2.38 log MPN/g, respectively, after 96 h of storage at 0 degrees C. However, populations of V. parahaemolyticus increased by 2.44 log MPN/g in shell stock oysters and by 1.64 og MPN/g in shucked oysters when stored at 15 degrees C for 60 h. Frozen storage was effective in inactivating V. parahaemolyticus. Populations of V. parahaemolyticus in shell stock and shucked oysters decreased from 5.46log MPN/g to 1.66 and 0.38 log MPN/g, respectively, after 75 days of storage at -30 degrees C. No V. parahaemolyticus cells were detected (<3 log MPN/g) in the shucked oysters after 60 days of storage at -18 degrees C. These results demonstrated that accumulation of V. parahaemolyticus in cultured C. plicatula increases as water temperature increases. Harvested C. plicatula should be stored at 5 degrees C or lower to control the hazard of V. parahaemolyticus.

[Determination of praziquantel residue in aquatic products using high performance liquid chromatography].

A sensitive and simple method for the determination of praziquantel residue in aquatic products has been established. The sample was extracted with ethyl acetate for three times (20 mL, 20 mL, 10 mL) and cleaned up with an LC-Si column. The extract was separated on a ZORBAX SB-C18 column (250 mm x 4.6 mm, 5 microm). Ultraviolet detection was performed at 214 nm. Acetonitrile-water (50: 50, v/v) was used as the mobile phase at a flow rate of 0.9 mL/min. The calibration curve of praziquantel in aquatic products at the concentration range of 0.02 - 20 mg/L was linear ( r = 0.999 98). The recovery of praziquantel was higher than 85%. The limit of detection was 10 microg/kg (S/N > 3).

[Study on decomposed products of trichlorfon in process of gas chromatographic analysis by mass spectrometry].

The decomposed products of trichlorfon in gas chromatographic analysis were identified by mass spectrometry (MS). After MS interpretation, three decomposed products, trichloroacetaldehyde, dimethyl phosphite and dichlorvos were identified. The effects of gas chromatographic conditions on decomposed products of trichlorfon, e. g. injection temperature, injection mode and oven ramp, were studied. The experiments showed that all of the factors have effects on decomposed products of trichlorfon, however, the injection temperature is the key factor to cause trichlorfon being decomposed. The higher the injection temperature is, the bigger the amount of trichlorfon being decomposed. When the injection temperature was raised from 150 degrees C to 250 degrees C, the remaining trichlorfon fell from 86% to 20%. Therefore, on-cold column injection mode gas chromatography or high performance liquid chromatography was recommended for exact quantification of trace trichlorfon.