Colorimetric determination of boric acid in prawns, shrimp, and salted jelly fish by chelate extraction with 2-ethyl-1,3-hexanediol.

Borate was directly chelate-extracted from foods with 5% 2-ethyl-1,3-hexanediol (EHD) in n-hexane-n-butyl acetate mixture (8 + 2), from which borate was selectively transferred into 1% NaOH, since EHD-chelated boron did not react with curcumin to develop color. Finally, an aliquot of the alkaline solution was acidified with HCl and reacted with curcumin in a rotary evaporator. Color development was increased by heating for 8 min at 80 degrees C under reduced pressure of 16 mm Hg. Frozen shrimp and prawns (peeled and with shells) and salted jelly fish were analyzed by the proposed method. Results were compared with the contemporary official method of Japan based on curcumin reaction on an incinerated sample. Over 90% of the boric acid was recovered by the proposed method when samples were fortified with 20 ppm boric acid. Recoveries were superior to those of the official method especially for shrimp and prawns with shells and salted jelly fish. Detection limit of boric acid is 1 ppm. Moreover, the method requires only about 1 hr for analysis of one sample, making it suitable for routine analysis.     

Determination of Organic Mercury in Biota, Plants and Contaminated Sediments Using a Thermal Atomic Absorption Spectrometry Technique

A simple, rapid procedure for the determination of organic mercury in sediments, plants and fish tissues has been developed and validated. Extraction and separation of organic mercury compounds from the sample matrix was achieved by an established procedure based on an acid leaching of the sample (H₂SO₄/KBr/CuSO₄), followed by extraction of the organic mercury halide with toluene and back-extraction with an aqueous solution of thiosulphate. Detection and quantification of mercury, in the liquid extracts, was made by atomic absorption spectrometry (AAS), following thermal decomposition of the sample. The method was evaluated using Certified Reference Material (CRM) BCR 463 (tuna fish), BCR 580 (estuarine sediment), IAEA-140TM (sea plant homogenate) and NRCC TORT-2 (lobster hepathopancreas). The recovery factors for organic mercury in all tested CRM were between 81–107%. The precision of the method has relative standard deviations of less than 10% for sediments and fish tissues and of less than 16% for plant material. The method was successfully applied to natural samples of sediments, plants, macroalgae and fish tissues collected from an estuarine ecosystem and could, therefore, be used for routine analyses.     

Determinations of residual furazolidone and its metabolite, 3-amino-2-oxazolidinone (AOZ), in fish feeds by HPLC-UV and LC-MS/MS, respectively

The antibacterial drug furazolidone belonging to the group of nitrofuran antibacterial agents has been widely used as an antibacterial and antiprotozoal feed additive for poultry, cattle, and farmed fish in China. During application a large proportion of the administered drug may reach the environment directly or via feces. Although the use of furazolidone is prohibited in numerous countries, there are indications of its illegal use. It is known that furazolidone can be rapidly metabolized to 3-amino-2-oxazolidinone (AOZ) in the body of the target organism. In this study, a total of 21 fish feed samples, including 17 commercial fish feeds from local markets in China (representing 15 different formulations) and 4 fish feeds obtained from Germany and Turkey, respectively, are analyzed to determine whether the drug is still illegally used or commercially available feeds are contaminated by this drug. High-performance liquid chromatography (HPLC) and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) methods have been implemented to determine furazolidone and its metabolite AOZ in fish feeds containing animal protein, respectively. An efficient and convenient cleanup method for the determination of furazolidone in fish feeds is developed, and a simple cleanup method for the determination of AOZ is used. Method recoveries for samples used were determined as 87.7-98.3% for furazolidone at two spike levels of 2.0 and 5.0 ng g-1 and as 95.6-102.8% for AOZ at spike levels of 0.4 and 0.8 ng g-1. Limits of detections were 0.4 ng g-1 for furazolidone and 0.05 ng g-1 for AOZ. The established methods are therefore suitable for the determination of furazolidone and its metabolite AOZ in fish feeds at trace contamination levels. Using the established methods, all fish feed samples have been proved to be furazolidone negative; however, AOZ is tested in 16 of 17 fish feeds obtained from local markets in the Hubei province of China, with a positive rate as high as 94.1%.     

Extraction and determination by gas chromatography of S,S,S-tri-n-butyl phosphorotrithioate (DEF) in fish and water

A simple, low-cost, fast method for the extraction and cleanup of DEF (S,S,S-tri-n-butyl phosphorotrithioate) from fish tissues and water samples was developed. The method combines extraction and cleanup in one step. The basis of the method is passing water samples or aqueous tissue homogenates containing DEF through a C-18 disposable cartridge. DEF is eluted from the cartridge by acetone or ethyl acetate. The eluates are analyzed by gas chromatography using a thermionic-specific detector. The method detects levels as low as 100 parts per trillion (ppt) in water samples; recovery efficiency from spiked fish tissues was greater than 95%. In addition, detectable levels of DEF were recovered from liver, brain, and muscle tissue of fish exposed to this compound. The method has a potential for use with other pesticides.     

基于光流法与特征统计的鱼群异常行为检测

鱼类群体行为的异常检测能够为鱼类健康监控与预警提供重要的方法和手段,对研究鱼类行为的机理,提升水产养殖过程中的信息化水平具有非常重要的意义。该文通过计算机视觉和图像处理技术,基于鱼群运动特征统计方法,对鱼群异常行为检测进行研究。利用Lucas-Kanade光流法得到目标鱼群的运动矢量,并对目标运动的行为特征进行统计,得到速度与转角这2个行为特征的联合直方图与联合概率分布。最后,在联合概率分布的基础上,基于标准互信息(normalized mutual information-NMI)和局部距离异常因子(local distance-based outlier factor-LDOF)2种方法对鱼群行为进行异常检测。试验结果表明,2种异常检测方法均达到99.5%以上的准确率。     

基于水下机器视觉的大西洋鲑摄食行为分类

根据鱼群摄食行为状态进行水产养殖精准投喂控制,是有效提高饵料利用率降低水体污染的关键技术。目前,大多数基于机器视觉的鱼类摄食行为研究都是在实验室对真实养殖环境进行模拟并采用水上摄像机获取数据,由于光照条件和养殖环境的影响,该数据无法反映大西洋鲑在实际生产状况下的摄食行为,因此应用范围有限。为解决此问题,该研究提出一种基于真实工厂化养殖环境的鱼类摄食行为分类算法。该算法使用水下观测方式并采用视频序列作为样本,首先利用变分自动编码器对视频序列样本进行逐帧编码以产生所有帧对应的高斯均值和方差向量,分别联立所有均值和方差向量得到均值特征矩阵和方差特征矩阵。然后将特征矩阵输入到卷积神经网络中,实现对鱼群的摄食行为分类。试验结果表明,在真实的工厂化养殖环境下,该研究所提出的方法综合准确率达到了89%,与已有的基于单张图像的鱼类摄食行为分类方法相比,综合准确率提高了14个百分点,召回率提高了15个百分点。研究结果可为基于鱼类摄食行为的鱼饵精准投喂控制提供参考。     

基于立体视觉的动态鱼体尺寸测量

获取渔业养殖鱼类生长态势的人工测量方法费时费力,且影响鱼的正常生长。为了实现水下鱼体信息动态感知和快速无损检测,该研究提出立体视觉下动态鱼体尺寸测量方法。通过双目立体视觉技术获取三维信息,再通过Mask-RCNN（Mask Region Convolution Neural Network）网络进行鱼体检测与精细分割,最后生成鱼表面的三维点云数据,计算得到自由活动下多条鱼的外形尺寸。试验结果表明,长度和宽度的平均相对误差分别在5%和9%左右。该研究满足了水产养殖环境下进行可视化管理、无接触测量鱼体尺寸的需要,可以为养殖过程中分级饲养和合理投饵提供参考依据。     

Determination and confirmation of melamine residues in catfish, trout, tilapia, salmon, and shrimp by liquid chromatography with tandem mass spectrometry

Pet and food animal (hogs, chicken, and fish) feeds were recently found to be contaminated with melamine (MEL). A quantitative and confirmatory method is presented to determine MEL residues in edible tissues from fish fed this contaminant. Edible tissues were extracted with acidic acetonitrile, defatted with dichloromethane, and cleaned up using mixed-mode cation exchange solid-phase extraction cartridges. Extracts were analyzed by liquid chromatography with tandem mass spectrometry with hydrophilic interaction chromatography and electrospray ionization in positive ion mode. Fish and shrimp tissues were fortified with 10-500 microg/kg (ppb) of MEL with an average recovery of 63.8% (21.5% relative standard deviation, n = 121). Incurred fish tissues were generated by feeding fish up to 400 mg/kg of MEL or a combination of MEL and the related triazine cyanuric acid (CYA). MEL and CYA are known to form an insoluble complex in the kidneys, which may lead to renal failure. Fifty-five treated catfish, trout, tilapia, and salmon were analyzed after withdrawal times of 1-14 days. MEL residues were found in edible tissues from all of the fish with concentrations ranging from 0.011 to 210 mg/kg (ppm). Incurred shrimp and a survey of market seafood products were also analyzed as part of this study.     

One-step extraction and cleanup procedure for determination of p,p'-DDT, p,p'-DDD, and p,p'-DDE in fish

A simplified method that combines extraction, partitioning, and cleanup in a single step for measuring p,p'-DDT and its metabolites in fish is described. Minced fish samples are emulsified with disodium hydrogen orthophosphate and trisodium citrate, ground with sodium sulfate, and eluted from a chromatographic column prepacked with alumina and silicic acid. The fats and fatty acids are solubilized and easily extracted from the tissues and retained by the column, while p,p'-DDT and its metabolites are quantitatively eluted with 40 mL n-hexane. The eluate is directly applied to a gas chromatographic column. Average recoveries of p,p'-DDT and its metabolites added to fish in vitro are 81%. The average coefficient of variation for recoveries of p,p'-DDT and its metabolites is less than 6.5% and the detection limit is 0.001 micrograms/g for p,p'-DDE, thus making this method very suitable for residue analysis.     

Determination of 49 organophosphorus pesticide residues and their metabolites in fish, egg, and milk by dual gas chromatography-dual pulse flame photometric detection with gel permeation chromatography cleanup

A new method for the quantitative determination of 49 kinds of organophosphorus pesticide residues and their metabolites in fish, egg, and milk by dual gas chromatography-dual pulse flame photometric detection was developed. Homogenized samples were extracted with acetone and methylene chloride (1 + 1, v/v), and then the extracts were cleaned up by gel permeation chromatography (GPC). The response of each organophosphorus pesticide showed a good linearity with its concentration; the linearity correlation was not less than 0.99. The detection limits (S/N = 3) of pesticides were in the range of 0.001-0.025 mg kg?1. The recovery experiments were performed by blank sample spiked at low, medium, and high fortification levels. The recoveries for fish, egg, and milk were 50.9-142.2, 53.3-137.2, and 50.3-139.4% with relative standard deviations (RSD, n = 6) of 2.3-24.9, 4.3-26.7, and 2.8-32.2%, respectively. The method was applied to detect organophosphorus pesticides in samples collected from the market, and satisfactory results were obtained. This quantitative method was highly sensitive and exact and could be applied to the accurate determination of organophosphorus contaminants in fish, egg, and milk.     

Gas-liquid chromatographics determination of Bayer 73 in fish, aquatic invertebrates, mud, and water

A gas-liquid chromatographic (GLC) method is described for determining residues of Bayer 73 (2-aminoethanol salt of 2',5-dichloro-4'-nitrosalicylanilide) in fish muscle, aquatic invertebrates, mud, and water by analyzing for 2-chloro-4-nitroaniline (CNA), a hydrolysis product of Bayer 73. Bayer 73 residues are extracted from fish muscle tissue, invertebrates, and mud with acetone-formic acid (98+2), and partitioned from water samples with chloroform. After sample cleanup by solvent and acid-base partitioning, the concentrated extract is hydrolyzed with 2N NaOH and H2O2 for 10 min at 95 degrees C. The CNA is then partitioned into hexane-ethyl ether (7+3) and determined by electron capture GLC. Average recoveries were 88% for fish, 82% for invertebrates, 82% for mud, and 98% for water at 3 or more fortification levels.     

Simple combustion-vapor trapping technique for determination of mercury in fish

A simple combustion train was constructed for analyzing mercury in fish samples. A nitric acid trap was used to capture the mercury vapors which were released later by adding a tin salt. The method is rapid, accurate, and reproducible and permits one person to analyze 40 samples daily. Sample matrix had no apparent effect on the recovery of mercury. Mean recoveries from fish samples spiked with 0.3 and 1.0 microgram mercuric chloride/g ranged from 97 to 105% with an average recovery of 101.5% and a standard deviation of 2.74%.     

自然水体中超声波标记鱼游动轨迹精密确定算法

针对鱼类关键生境位置确定的应用需求,该文提出了一套适用于自然水体的超声波标记鱼定位算法,解决了标记鱼定位以及存在粗差观测值,即水听器记录的超声波信号接收时间存在错误情况下算法的抗干扰性。宜昌黄柏河的实测结果表明,基于现有1 ms级精度的水听器,可在自然水体中获得2.15 m精度的信号标记鱼三维游动轨迹。如因气泡、遮挡等因素对水听器观测数据引入粗差,当粗差量级在10 m以上,该方法可接近100%探测出是否存在粗差。当粗差观测值在3个以内时,该方法的探测成功率可达84.3%以上,3个以上时粗差探测成功率明显下降,5个及以上,即粗差观测值个数占观测值总数的比例大于31.25%时,基本只能探测出观测数据中存在粗差而无法有效确定粗差。该研究可为渔业增殖、鱼类栖息地保护、鱼类洄游通道等研究提供参考。     

Usefulness of near-infrared reflectance (NIR) spectroscopy and chemometrics to discriminate fishmeal batches made with different fish species

Near-infrared reflectance (NIR) spectroscopy combined with chemometrics was used to identify and authenticate fishmeal batches made with different fish species. Samples from a commercial fishmeal factory (n = 60) were scanned in the NIR region (1100-2500 nm) in a monochromator instrument in reflectance. Principal component analysis (PCA), dummy partial least-squares regression (DPLS), and linear discriminant analysis (LDA) based on PCA scores were used to identify the origin of fishmeal produced using different fish species. Cross-validation was used as validation method when classification models were developed. DPLS correctly classified 80 and 82% of the fishmeal samples. LDA calibration models correctly classified >80% of fishmeal samples according to fish species The results demonstrated the usefulness of NIR spectra combined with chemometrics as an objective and rapid method for the authentication and identification of fish species used to manufacture the fishmeal.     

Spectrofluorometric determination of histamine in fish and meat products

A method has been developed for spectrofluorometric determination of histamine in fish and meat products. After a perchloric extract is obtained from samples, histamine is extracted with n-butanol and transferred to hydrochloric acid. Finally, histamine is subjected to a condensation reaction with o-phthalaldehyde (OPT). The method was tested for lack of interference from other amines. Precision of the method in fish products was 6.60% CV; recovery was 96.50%. In meat products, precision was 5.42% CV; recovery was 96.20%. By analysis of variance (P = 0.05), no significant statistical differences were found for recovery values vs histamine content in both foods.     

Screening of tetracycline residues in fish muscles by CCD camera-based solid-surface fluorescence

A methodology for the screening of tetracyclines (TCs), including tetracycline (TC), oxytetracycline (OTC), and chlorotetracycline (CTC), in different fish muscle matrices has been proposed. This method was based on in situ fluorescent derivation of TCs, transferring weakly fluorescing TCs to highly fluorescent species, on alkaline-activated solid silica gel G plates (SGGPs). By coupling solid-surface fluorescence (SSF) with charge-coupled device (CCD) camera imaging, a CCD camera-based SSF (CCD-SSF) methodology has been developed. Calibration curve, repeatability, selectivity, limit of detection (LOD), and limit of quantification (LOQ) have been explored for evaluating the performance of the method itself. Linear calibration curves were obtained over a range of 0.20-1.0 ng/spot for all three TCs. The LODs, defined as 3sigma, for TC, OTC, and CTC were 0.14, 0.15, and 0.16 ng/spot, respectively. The trueness of method was validated by HPLC, and no significant difference between CCD-SSF and HPLC was found, on a basis of 95% confidence level. By spiked recovery studies, a linear calibration curve ranging from 20 to 300 microg/kg of TC in fish muscle samples with a correlation coefficient (R 2) equal to 0.994 was obtained. The total average recovery for TC in fish muscle samples from six different fish matrices, fortified with TC at 50, 100, and 200 microg/kg levels, was 75.7% with average relative standard deviations (RSDs) ranging from 2.0 to 7.7%. RSDs ranged from 2.5 to 5.8% and from 5.2 to 7.6% for in-day and interday repeatability, respectively. The detection and quantification limits in fish muscle matrices were 16 and 53 microg/kg of TCs, respectively. The newly developed CCD-SSF method has been applied to the screening of the TC residues in fish muscle samples. The method has been demonstrated to bear some advantages, such as its simplicity, high throughput, low cost, use of fewer pollutants, and reasonable sensitivity.     

饲料蛋白质水平对刺鲃幼鱼的生长、胴体营养组成及消化酶活性的影响

配制蛋白质水平20%~50% 的7种等能半精制实验饲料,饲养平均初始体重为1.26±0.02 g 的7组3重复的刺鲃(Barbudes caldwell)幼鱼8周,研究刺鲃对饲料蛋白的需求量。结果显示,饲料蛋白水平对实验鱼的成活率、脏体比和肝体比等均无显著影响(P >0.05),而对鱼体终重有显著影响(P <0.05);鱼体增重和生长比速均随饲料蛋白水平从20%升高到35%而不断提高,但饲料蛋白水平进一步提高,则鱼体增重和生长比速均无显著差异(P >0.05)。饲料蛋白水平30%～50%的5个实验组的饲料效率无显著差异(P>0.05), 但均显著高于饲料蛋白为20%和25%的2个实验组(P <0.05)。蛋白质效率(PER)与饲料蛋白水平(x)呈负相关(PER= 3.006－0.03251x, R =0.9366)。饲料蛋白水平对鱼体的胴体水分、粗蛋白和粗灰分的含量无显著影响(P > 0.05);胴体脂肪含量随饲料蛋白水平(x)的升高而不断降低(L=8.2169－0.0458x,R =0.8551)。实验鱼肝胰脏蛋白酶活性则在组间无显著差异(P > 0.05)。肠蛋白酶和肝胰脏淀粉酶活性均随着饲料蛋白水平的升高而呈现先逐渐升高,且升高到一定程度后又呈逐渐下降的趋势。肠淀粉酶活性(IAA)与饲料蛋白水平(x)呈负相关关系(IAA = 84.625－0.9147x, R =0.8463,)。以鱼体增重为指标(y),与饲料蛋白水平(x)拟合折线模型进行回归分析,估算出饲料蛋白质适宜水平为占干饲料的34%。     

Simple confirmatory method for the determination of erythromycin residues in trout: a fast liquid-liquid extraction followed by liquid chromatography-tandem mass spectrometry

In recent years, erythromycin has received considerable attention for its therapeutic efficacy against some bacterial kidney diseases in aquaculture and, therefore, suitable and sensitive analytical methods to monitor erythromycin residues in fish are required. A fast sample treatment followed by an LC-ESI-MS/MS method is described for the purification, identification, and quantification of erythromycin A residues in fish. After two extractions with acetonitrile, samples were defatted with n-hexane, filtered, and analyzed by tandem mass spectrometry. Three characteristic transition reactions (m/z 734 --> 716, 734 --> 576, and 734 --> 558) in multiple reaction monitoring were tested for the determination and confirmation of erythromycin A. The method was in-house validated through the determination of precision, accuracy, specificity, stability, calibration curve, decision limit (CCalpha), and detection capability (CCbeta), in accordance with European Commission Decision 657/2002. The coefficients of variation ranged from 1.8 to 9.4% and from 7.5 to 10.9% for intra- and interday repeatability, respectively. Recovery data were also satisfactory, with values varying from 85 to 97%. The method was specific, stable, and robust enough for the required purposes. The calibration curve showed a good linearity in the whole range of the tested concentrations (0-1000 microg kg(-1)) with a correlation coefficient (r2) equal to 0.9956. CCalpha and CCbeta were found to be 220 and 238 microg kg(-1), respectively.     

Uptake from water,depuration and tissue distribution of110mAg in a freshwater fish,Cyprinus carpio L.

Radionuclide accumulation was studied on a group of 20 fish maintained at 20±1 °C in spring water containing 30 Bq mL^?1 of^110mAg. Because of a very significant radionuclide adsorption onto available surfaces and in order to simulate chronic exposure, the water was completely renewed three times a week. Accumulation of^110mAg was slow, with an estimated time to maximum contamination level of 180 days. The concentration factor, calculated from the ratio of the integrals of the curves representing the radionuclide concentration in filtered water and in whole fish reached a maximum value of 106 (w.w.). After 41 days exposure, 10 fish were placed in non-labelled water, renewed daily, to follow^110mAg depuration. A two-compartment exponential model was fitted to the depuration data. The corresponding radionuclide half-lives were relatively short, 0.8 (Tb₁) and 30 days (Tb₂), and after 6 weeks the fish retained about 30% of their initial^110mAg content. The liver and digestive tract retained most^110mAg. Although they only represented respectively about 4 and 10% of the total body mass, these two organs accounted for 30–40% of the total radionuclide body burden.     

Identification of O,O-dialkyl-S-methylphosphorodithioate residues in fish

O,O-Dialkyl-S-methylphosphorodithioates were found in Mississippi River buffalo fish caught near several chemical plants and oil refineries in Hartford and Wood River, IL. These chemicals, which have not been previously recognized as environmental or food contaminants, were identified and quantitated by a procedure similar to the AOAC multiresidue method for organochlorine and organophosphorus pesticides, using gas chromatography (GC) with flame photometric detection (FPD). The key to their identification was a GC/FPD retention time pattern that was virtually the same as that for the diazomethane reaction products of a commercial zinc dialkyl dithiophosphate motor oil additive. GC/mass spectrometry (MS) showed that the compound producing the largest GC/FPD peak contained butoxy groups. The identification of this compound as O,O-di(2-methylpropyl)-S-methyl-phosphorodithioate (Compound C) was confirmed by GC/MS analysis by comparison with the authentic material. The buffalo fish contained 0.15 ppm Compound C and 0.5 ppm total O,O-dialkyl-S-methylphosphorodithioates. Subsequent analyses of fish from other areas showed that these contaminants were not limited to the Hartford-Wood River area. Lower residue levels of Compound C, ranging from 0.01 to 0.05 ppm, were found in fish from the Mississippi River at Sauget, IL, and from the Delaware River and Newark Bay in NJ.