高效液相色谱法检测海水养殖环境中喹诺酮类药物残留

建立了一种养殖海水中诺氟沙星、环丙沙星和恩诺沙星3种喹诺酮类抗生素残留量的高效液相色谱-荧光测定方法。水样经稀盐酸调pH后经HLB固相萃取柱富集、净化,用外标法定量。结果表明,诺氟沙星、环丙沙星和恩诺沙星检出限分别是2、1和1μg/L,定量限分别是6.6、3.3和3.3μg/L,添加浓度为10、20和50μg/L时,3种物质的回收率为71.9%～85.3%,批内变异系数≤10%,批间变异系数≤7%。利用该方法对黄海沿岸部分养殖场及近岸海水(北起36°40.303′N,121°8.939′E;南至35°39.36′N,119°52.433′E)进行分析,诺氟沙星、环丙沙星、恩诺沙星的检出质量浓度分别为6.20～982、55.2和11.6～55.4μg/L。实验结果表明,该方法灵敏度高,重现性好,适用于养殖海水中诺氟沙星、环丙沙星和恩诺沙星的检测。     

广州市番禺区淡水养殖水产品中氟喹诺酮类药物残留现状分析及对策

为了解广州市番禺区淡水养殖水产品中氟喹诺酮类抗生素的残留量，并对存在的风险点进行预警。利用高效液相色谱-串联质谱法对鱼肉中的7种氟喹诺酮类(诺氟沙星、培氟沙星、洛美沙星、氟罗沙星、氧氟沙星、环丙沙星、恩诺沙星)残留量进行测定。结果表明，2022年共抽检191个养殖户的315批次水产品，共检出61个养殖户共75批次水产品氟喹诺酮类药物残留，未检出超过限量标准的样品。养殖户的检出占比为31.94%,水产品总体检出率为23.81%;主要检出恩诺沙星75批次、环丙沙星16批次，其他氟喹诺酮类未检出。结论:广州市番禺区淡水养殖水产品氟喹诺酮类药物残留主要为恩诺沙星和环丙沙星，残留情况较普遍，应加强对这两类药物的监测。     

高邮养殖区罗氏沼虾中多种抗生素的残留与膳食暴露评估

对高邮市9个养殖区的45个罗氏沼虾(Macrobrachium rosenbergii)样品中的4类(磺胺类、喹诺酮类、四环素类及大环内脂类)39种抗生素残留进行快速筛查验证,分析残留现状与健康风险。样品罗氏沼虾经酸化乙腈和乙二胺四乙酸二钠提取浓缩后,再经正己烷去脂,应用液相色谱-三重四级杆复合线性离子阱质谱(LC-Q-TRAP/MS)技术,采用内标法定量和分段时间多反应监测-信息关联采集-增强离子扫描的方式(sMRM-IDA-EPI),进行定性和定量分析。结果表明,9个养殖区抗生素残留中磺胺类、四环素类和大环内酯类均未检出,喹诺酮类抗生素洛美沙星、培氟沙星、氧氟沙星和诺氟沙星4种禁用兽药均未检出,仅有环丙沙星、恩诺沙星及达氟沙星被检出,检出率分别为44.4%、82.2%和51.1%,总体合格率为93.3%。达氟沙星含量较小,平均含量范围在ND(未检出)～9.00μg/kg。恩诺沙星和环丙沙星总量范围在N.D.～157.00μg/kg,个别养殖区组(SQZ组和SDZ组)内检出率100%,其中3个样品超过限量值100μg/kg。使用每日允许摄入量(acceptable daily intake,ADI)和估计日摄入量(estimated daily intake,EDI)评价膳食安全,喹诺酮类(恩诺沙星和环丙沙星)、达氟沙星暴露风险(EDI/ADI)值在1.66×10~(-5)～8.06×10~(-3)范围内,表明9个养殖区罗氏沼虾虾体膳食暴露评估风险水平在可接受范围内,对人体造成健康风险的可能性较小。研究认为,高邮养殖区罗氏沼虾中抗生素污染程度较轻,罗氏沼虾中喹诺酮类药物残留量较高,经膳食暴露评估尚在安全范围内,建议进一步加强高邮地区罗氏沼虾等养殖水产品中喹诺酮类等抗生素的监控管理。[中国渔业质量与标准,2019,9(6):23-30]     

水产常用抗菌药物对锯缘青蟹酚氧化酶的影响

研究了喹诺酮类等17种常见抗菌药物对青蟹酚氧化酶的影响。试验结果表明,药物质量浓度为500μg/ml时,所有抗菌药物对青蟹酚氧化酶(PO)均有明显抑制作用;药物质量浓度为5~10μg/ml时(常规使用剂量),庆大霉素、卡那霉素和丁胺卡那霉素等5种药物的PO抑制率为10%~25%;诺氟沙星、环丙沙星和氧氟沙星等6种药物的PO抑制率为5%~10%,抑制作用较弱;恩诺沙星和红霉素等4种药物的PO抑制率均小于5%,对青蟹PO无明显影响;低剂量阿维菌素和氟苯尼考对青蟹有免疫促进作用。     

上海市售鳜渔药残留现状的调查研究

为了解上海市场销售鳜(Siniperca chuatsi)渔药残留现状,于2018年4月-2019年3月,对上海市批发市场销售的72份鳜肌肉样,采用高效液相色谱法,分别测定孔雀石绿(MG)、硝基呋喃(NFs)、喹诺酮(4-Qs)以及亚甲基蓝(MB)四大类共10种药物。结果显示:孔雀石绿、隐色孔雀石绿以及呋喃唑酮、呋喃它酮无检出,呋喃西林、呋喃妥因检出样占样本量的50%,喹诺酮类占76.39%,亚甲基蓝占15.3%;呋喃西林的检出浓度为(0.49±0.03)μg/kg、呋喃妥因(3.51±2.27)μg/kg,诺氟沙星(25.52±10.36)μg/kg、环丙沙星(20.98±6.85)μg/kg、恩诺沙星(27.95±4.17)μg/kg、亚甲基蓝(120±14)μg/kg;呋喃唑酮类主要集中在10月份到次年4月份之间,喹诺酮类与亚甲基蓝则主要集中在4月份到10月份之间,环丙沙星全年可检出。结果表明:市售鳜肌肉中仍有部分禁用药物和限用渔药的残留,且不同药物残留表现出一定的季节性差异。     

模拟生态系统中恩诺沙星在沉水植物体内蓄积、代谢和消除规律

为评价沉水植物对去除养殖尾水中残留抗菌药的作用,改进了QuEchERS前处理方法,建立了沉水植物中恩诺沙星和环丙沙星含量HPLC检测方法,采用模拟生态研究了恩诺沙星残留在沉水植物体内蓄积、代谢和消除规律。优化了不同提取液、萃取盐和提取时间对回收率的影响,考察了不同净化剂的净化效果。以外标法进行定量,沉水植物中恩诺沙星和环丙沙星回收率分别为79.60%～110.15%和57.26%～102.00%,检测限分别为0.003μg·g^-1和0.005μg·g^-1。在模拟生态系统中,以终浓度200μg·L^-1的恩诺沙星单次泼洒用药后,苦草(Vallisneria natans)、伊乐藻(Elodea nuttallii)和轮叶黑藻(Hydrilla verticillata)均在24 h吸收恩诺沙星至最大值,分别为0.82μg·g^-1、0.86μg·g^-1和2.34μg·g^-1;采用梯形法计算曲线下面积,分别为192.06(μg·g^-1)·h、...     

高效液相色谱法同时测定水产品中四种氟喹诺酮类药物残留前处理条件的优化

建立了同时测定水产品中氧氟沙星、诺氟沙星、环丙沙星、恩诺沙星4种氟喹诺酮类药物残留量的高效液相色谱方法,并对提取和净化等前处理条件进行了优化。目标化合物采用高效液相色谱-荧光检测器检测,外标法定量。对草鱼、鳙鱼、大黄鱼、虾进行检测分析,添加4、10、100、1000μg/kg浓度水平时,回收率为68.5%~106.6%,相对标准偏差0.80%~9.16%,方法的检出限为1.5μg/kg,定量限为4μg/kg。     

基于高效液相色谱-四级杆-飞行时间质谱技术筛查渔业水域环境中抗生素残留方法的建立与应用

应用高效液相色谱—四级杆—飞行时间质谱(HPLC-Q-TOF-MS)技术建立水中常用的42种抗生素残留快速筛查方法。采用固相萃取富集净化的方法进行样品预处理,目标药物经液相色谱仪C18反相柱分离,1%甲酸乙酸铵水溶液—甲醇溶液为流动相梯度洗脱,利用正离子源全扫描模式进行检测,并将建立的方法应用于浙江省内主要渔业环境水质样品抗生素残留的筛查工作。渔业水域环境中42种抗生素定量限(LOQ,S/N=10)为0.010~0.120μg/L。3个水平加标的平均回收率为60.7%~97.1%,相对标准偏差(RSD)为0.58%~8.01%。该方法快速简便,灵敏度高,对抽取的60批次样品进行检测,共检出磺胺甲鰁唑、磺胺间甲氧基嘧啶、磺胺吡啶、磺胺氯哒嗪、恩诺沙星5种药物,含量为25.6~106.7 ng/L。本实验建立的渔业水质中抗生素药物的高效液相色谱—四级杆串联飞行质谱分析方法,灵敏、准确、定性能力强,可以成功应用于渔业环境水质样品抗生素残留的筛查工作。     

液相色谱法检测鸡蛋中四种氟喹诺酮类药物残留前处理方法探讨

建立了高效液相色谱法测定鸡蛋中环丙沙星、达氟沙星、恩诺沙星、沙拉沙星4种氟喹诺酮类药物残留的方法。样品经过磷酸盐缓冲液提取,水饱和正己烷去除蛋白质和脂类物质。目标化合物采用高效液相色谱-荧光检测器检测,外标法定量,结果表明,4种氟喹诺酮类残留药物能在15 min内得到有效分离,峰形对称。方法检出环丙沙星、恩诺沙星、沙拉沙星为10μg/kg,达氟沙星为2μg/kg,回收率在83.2%~97.3%,相对标准偏差在3.6%~8.7%。     

高效液相色谱-串联质谱法检测3种渔业环境基质中的孔雀石绿及隐性孔雀石绿残留

为了准确评估渔业生态环境中的孔雀石绿(MG)及其代谢产物隐性孔雀石绿(LMG)的残留状况,建立了3种渔业环境基质(水体、底泥和底泥-水体混合物)中MG和LMG的高效液相色谱-串联质谱(HPLC-MS/MS)检测方法。通过考察不同前处理方法对不同基质中MG和LMG回收率的影响,优化仪器性能,确定了色谱和质谱分析条件。具体方法是:水体过滤后采用PRS固相萃取柱进行净化、富集;底泥采用乙腈-二氯甲烷(1∶1,v/v)提取,再旋蒸、富集;底泥-水体混合物用二氯甲烷提取后,采用PRS固相萃取柱净化、富集。采用Thermo C18色谱柱对待测物进行分离,以乙腈-0. 2%乙酸铵(1∶1,v/v)为流动相洗脱,电喷雾-多反应正离子监测模式监测,内标法定量。结果表明,3种不同基质中的MG和LMG在1～8 ng/m L范围内线性显著,其相关系数r~2值大于0. 999;加标回收率分别为90. 0%～104. 2%、79. 9%～90. 3%和74. 1%～86. 3%,相对标准偏差为3. 3%～5. 8%、4. 9%～8. 6%和3. 2%～8. 3%; MG的检出限(LOD,S/N=3)分别为0. 24 ng/L、0. 02μg/kg和0. 06μg/kg,定量限(LOQ,S/N=10)分别为0. 79 ng/L、0. 07μg/kg和0. 21μg/kg; LMG的检出限(LOD,S/N=3)分别为1. 14 ng/L、0. 17μg/kg和0. 12μg/kg,定量限(LOQ,S/N=10)分别为4. 72 ng/L、0. 56μg/kg和0. 39μg/kg。该法可应用于渔业环境中MG及LMG的定性定量检测,具有较好的实用性。     

盐度和漂白粉对东方新糠虾的急性胁迫影响

在水温(26.5±0.5)℃的条件下,探究了盐度和漂白粉对体长为(0.66±0.07)cm的东方新糠虾(Neomysis orientalis)的急性毒性作用。实验结果表明:东方新糠虾适宜的盐度范围较广,对低盐的耐受能力更强,其最适盐度范围为15～25。漂白粉对东方新糠虾在24、48、72 h的半致死浓度分别为2.639、1.149、0.812 mg/L,安全浓度为0.065 mg/L。     

水温对额尔齐斯河银鲫幼鱼耗氧率及窒息点的影响

为鱼类养殖生产和苗种运输过程中溶解氧的科学调控提供理论依据,该试验采用密闭呼吸室法测定了20、22、24、26、28、30℃水温下额尔齐斯河银鲫幼鱼的耗氧率和窒息点。结果表明,水温在20～30℃时,额尔齐斯河银鲫幼鱼的耗氧率随水温的升高而显著上升(P<0.05)。水温为20℃时耗氧率最低,为0.263 mg/(g·h);水温为30℃时耗氧率最高,为0.571 mg/(g·h)。耗氧率与水温的拟合方程为:y=0.0093x^2-0.0027x+0.2604(R^2=0.9896)。额尔齐斯河银鲫幼鱼的窒息点在20～30℃水温下,随水温的升高而上升,其中水温为20℃时窒息点最低,为0.947 mg/L;水温为30℃时窒息点最高,为1.713 mg/L。水温和窒息点的回归关系方程式为:y=0.0271x^2-0.0629x+1.0452(R^2=0.9116)。     

甲氰菊酯对尼罗罗非鱼组织乙酰胆碱酯酶、谷丙转氨酶和谷胱甘肽活性的影响

用急性染毒法研究6种浓度(0、0.4、0.6、0.8、1.0、1.5 μg/L)的甲氰菊酯对(34.89±9.99)g的尼罗罗非鱼(Tilapia nilotica)分别处理6、12、24、48、96 h后,其组织内乙酰胆碱酯酶(AChE)、谷丙转氨酶(GPT)和谷胱甘肽( GSH)的动态变化.结果显示:AChE活性在...     

Fatty acid composition of New Zealand green-lipped mussels, Perna canaliculus: Implications for harvesting for n-3 extracts

Marine bivalves offer a potentially important source of long-chained polyunsaturated fatty acids (PUFA) for human health supplements. Lipid extracts from individual New Zealand green-lipped mussels (NZGLM) were analyzed as fatty acid methyl esters (FAME) by gas chromatography to assess geographical and seasonal differences between large (86 ± 1 mm) male and female and small (44 ± 1 mm) mussels. PUFAs dominated in spring and summer, comprising ∼ 50% of total fatty acids. Moreover, the commercially important n-3 fatty acids, 20:5n-3 (eicosapentaenoic acid, EPA) and 22:6n-3 (docosahexaenoic acid, DHA) together accounted for 70-79% of total PUFAs in spring and summer. During winter there was a marked decrease in condition and total n-3 PUFAs and a concomitant increase in saturated fatty acids in mussels, suggesting they had already spawned, had increased metabolic demands and limited PUFA-rich phytoplankton as food. While total n-3 content was not significantly different, there were geographical differences in individual n-3 fatty acids. Mussels collected from the cooler waters of Stewart Island had greater levels of 20:5n-3 (EPA), while those collected in Marlborough had greater concentrations of 22:6n-3 (DHA), which was attributed mainly to differences in phytoplankton composition. Total n-3 content and the condition index varied seasonally with greater concentrations of n-3 PUFAs, especially EPA, recorded in large mussels in spring, coincident with spring diatom blooms. Total PUFA levels and condition indices remained high in summer. There was no significant difference in condition indices, total n-3 content, DHA or EPA levels between large male and female mussels. Conversely, large mussels had significantly greater amounts of n-3 PUFAs than small mussels at Marlborough Sounds, while small mussels had marginally greater total n-3 concentrations than large mussels at Stewart Island. Taken together, these results suggest that the NZGLM offers a potentially important source of n-3 PUFAs for human health supplements. Our findings suggest that optimal harvesting conditions occur in spring when mussel condition and n-3 content peak for large mussels. Although DHA and EPA levels varied geographically, total n-3 content was not significantly different between sites, which implies that harvesting mussels for n-3 extracts would be driven more by logistical considerations.     

30日龄翘嘴鳜形态性状与体质量之间的关系

为确定适于翘嘴鳜(Siniperca chuatsi)人工选育的理想测度指标,随机选取200尾30日龄翘嘴鳜,测定其体质量(y)、全长(x1)、体长(x2)、吻长(x3)、眼径(x4)、体厚(x5)、体高(x6)、头长(x7)、尾柄长(x8)和尾柄高(x9)共10个指标,采用相关分析、通径分析和多元回归分析,探讨了30日龄翘嘴鳜形态性状与体质量之间的关系。结果表明,30日龄翘嘴鳜体质量变异系数最大(达26.73%),各形态性状与体质量的相关系数均达到极具统计学意义水平(P<0.01)。模型拟合结果显示,体高、体长、尾柄长和体厚4个形态性状与体质量的最优模型决定系数R2达0.935。这4个形态性状中,体高对体质量的直接作用(0.452)和决策系数(0.634)均为最大,说明体高为主要决策变量。利用逐步回归分析方法,建立的多元回归方程为:y=-0.672+0.053x6+0.019x2+0.015x8+0.019x5。该研究为翘嘴鳜选育工作提供了理论依据和技术参数。     

蟹塘生态调控技术对“淤泥-水界面”N,P交换通量的影响

调节pH值、加钙和增氧是常用的蟹塘生态调控技术,为明确其对水质和N,P养分交换的影响,通过实验室模拟并测定了水体N,P含量及"淤泥-水界面"交换通量。结果表明:3种措施对水体N,P均有显著影响,且增氧对水体N,P的调节效应显著强于加钙和调节pH。增氧96 h后水体TN和NO_3^--N含量分别是不增氧处理的1.68和7.43倍,同时增氧削减了水体NH4+-N含量至试验结束,增氧处理NH_4^+-N含量平均降低70%～75%。"淤泥-水界面"N,P交换通量表明,增氧后TN交换通量平均提高2.47～3.74 mg·m^2/h,而对TP交换通量均无显著影响,平均仅为-0.1 mg·m^2/h。     

温度对乌苏里拟鲿受精孵化及仔鱼生长的影响

为探究乌苏里拟鲿胚胎发育及仔鱼生长的最适温度,采用单因子试验设计研究了不同温度(24、26、28和32℃)对乌苏里拟鲿胚胎受精、孵化及仔鱼存活、生长的影响。结果表明:高温组(32℃)的乌苏里拟鲿受精率、存活率均较其他组显著降低(P<0.05);低(24℃)、高温组(32℃)的孵化率显著低于对照组(26℃)和中温组(28℃)(P<0.05);高温组(32℃)5—20日龄的乌苏里拟鲿生长缓慢,但20—30日龄生长加速,到30日龄时与对照组差异不具统计学意义。利用二次方程对乌苏里拟鲿仔鱼(0—30d)的生长率(AGR)进行拟合分析,得到拟合方程为:AGR=0.0225T-0.0004T2-0.2988通过求导推算极值的方法,得出乌苏里拟鲿的最适生长发育温度为28.13℃。     

Diel pattern of haematocrit, serum metabolites, osmotic pressure, electrolytes and thyroid hormones in sea bass and sea bream

The diel pattern of haematocrit, serum metabolites, electrolytes and thyroid hormones are described in sea bass, Dicentrarchus labrax, and sea bream, Sparus auratus, held under 12L:12D light cycle (0600–1800 h light). No diel rhythmicity in haematocrit, total proteins, potassium and chloride was observed. Maximum values of glucose, osmotic pressure and sodium were recorded at dusk (1800 h) in both species, irrespective of differences in the size of fish and feeding regime used. Maximum and minimum triiodothyronine values were found during the scotophase in bass and during the photophase in bream. Maximum thyroxine levels occurred at 1400 h (bass) and 1800 h (bream) and minimum during the scotophase, in both species. The light/dark alternation seems to be an important synchronizer of the observed diel changes. Results are discussed in relation to the timing of feeding and the general activity of the fish.     

Genetic linkage map of the pearl oyster,Pinctada martensii (Dunker)

Genetic linkage maps were constructed with amplified fragment length polymorphism (AFLP) and microsatellite markers for the pearl oyster, Pinctada martensii (Dunker), the main bivalve used for marine pearl production in Asia. Twenty‐four AFLP and 84 microsatellite primer pairs were used for linkage analysis in a full‐sib family with two parents and 78 offspring. Of the 2357 AFLP fragments generated, 394 (16.7%) were polymorphic and segregating. Most (340 or 86.2%) of the markers segregated according to expected Mendelian ratios. Female and male linkage maps were constructed using 230 and 189 markers, including 15 and 10 microsatellites respectively. The female map consisted of 110 markers in 15 linkage groups, covering 1415.9 cM, with an average interval of 14.9 cM. The male map consisted of 98 markers in 16 linkage groups, with a total length of 1323.2 cM and an average interval of 16.1 cM. When unlinked doublets were considered, genome coverages were 78.5% for the female and 73.5% for the male map. Although preliminary, the genetic maps constructed here should be useful for future linkage and quantitative trait loci mapping efforts.     

Oral bovine serum albumin immune‐stimulating complexes improve the immune responses and resistance of large yellow croaker (Pseudosciaena crocea,Richardson, 1846) against Vibrio alginolyticus

This study aimed to investigate effects of bovine serum albumin immune‐stimulating complexes (BSA ISCOMs) on immune‐related genes expression, serum nonspecific immunity and disease resistance of large yellow croaker (Pseudosciaena crocea). Fish were fed diets containing 3.5 ml of BSA ISCOMs per kg feed (experimental group) or 3.5 ml of phosphate‐buffered saline per kg feed (control group) for 1 week. The liver, spleen, head‐kidney tissues were sampled for determining gene expression of myxovirus‐resistant protein (Mx), major histocompatibility complex class II alpha chain (MHC II α), tumour necrosis factor‐alpha (TNF‐α) and interleukin‐10 (IL‐10) 30 and 90 days after feeding. Also, blood samples were collected for determining activities of serum superoxide dismutase (SOD), interferon alpha (IFN‐α), TNF‐α and alkaline phosphatase (ALP). TNF‐α and MHCⅡα gene expression in the liver, spleen, head‐kidney, as well as IFN‐α, TNF‐α and ALP activities in the serum, of experimental fish were significantly higher 30 days after feeding; while only TNF‐α and MHC II gene expression in the head‐kidney remained upregulated 90 days after feeding. The cumulative mortality of the experimental fish was significantly lower than control. This study indicated that BSA ISCOMs improved the immune response and induced protective immunity in large yellow croaker.