超声波法制备高黏度的壳聚糖

以甲壳素为原料,用超声波法制备高黏度的壳聚糖。研究超声波的温度、超声的时间以及制备壳聚糖时浓碱的浓度3个不同条件对壳聚糖黏度的影响。结果表明,用超声波制备高黏度壳聚糖的最佳反应条件是:反应温度为50℃,反应时间为3 h,NaOH溶液的浓度为50%,可以得到黏度为2886.7mpa.s、脱乙酰度达97.17%的壳聚糖。将得到的高黏度壳聚糖制成膜,用于染料废水的处理,15 h后,壳聚糖膜对活性橙的脱色率就可达到99.5%。     

不同强度超声波辅助浸浴法导入草鱼嗜水气单胞菌疫苗免疫效果评价

采用不同强度的超声波处理,研究鱼类免疫效应和疫苗导入效果,从而优化最适超声波处理强度,以便应用于渔业实践中。本研究中,在56kHz的超声频率下,采用不同超声强度(0、83、166、250、333mW/cm2),用以辅助浸浴法导入草鱼嗜水气单胞菌疫苗。结果显示,用166、250mW/cm2的超声波强度,免疫14d后,超声处理组溶菌酶活性和白细胞吞噬活性最高,显著高于对照组(P0.05);免疫28d后,血清凝集效价达到最高,显著高于对照组(P0.05)。超声波强度为166、250mW/cm2时,超声波处理组的相对免疫保护率高于对照组和其他处理组,分别达到66.5%和83.5%。本研究表明,用166mW/cm2和250mW/cm2的超声波强度处理后,鱼体免疫效价显著提高,表明该强度可作为适宜的超声波处理强度。实际生产中可采用166~250mW/cm2超声波强度处理,以辅助浸浴法有效导入草鱼细菌性疫苗,研究结果对于渔业生产应用和示范推广将提供重要指导。     

近江牡蛎的超声波辅助自溶及酶解条件研究

超声波和蛋白酶可以破坏蛋白质结构,提高短肽的得率。本试验以近江牡蛎为原料,在适度水解、控制挥发性盐基氮在合理范围的前提下,以短肽含量为主要目标,首先通过研究超声波功率、时间对牡蛎自溶酶解的影响,优化超声波辅助酶解工艺,然后采用单因素试验研究碱性蛋白酶对超声波—自溶酶解液酶解效果的影响。结果显示,440 W超声波处理牡蛎10min后自溶酶解,短肽含量为(88.75±1.16)mg/mL,是对照组的2.12倍;水解度为(30.38±0.05)%;挥发性盐基氮为(79±2)μg/g。采用碱性蛋白酶对超声波-自溶酶解液进行进一步酶解,优化的酶解条件是:加酶量5kU/g,酶解温度55℃,酶解时间4h,此时短肽含量达到100mg/mL以上,水解度为(35.95±0.05)%;挥发性盐基氮为(178±6)μg/g,处在可接受范围。试验结果表明,超声波预处理后自溶,再进行外源蛋白酶酶解可显著提高酶解效率。     

超声波优化辅助盐渍海蜇脱铝工艺

为探究超声波辅助盐渍海蜇脱铝工艺的条件,实验以盐渍海蜇为原料,采用电感耦合等离子体质谱法（ICP-MS）进行检测,其铝残留量为考察指标,通过单因素实验研究复合保鲜液浓度、超声功率、超声频率以及超声时间对盐渍海蜇脱铝效果的影响,根据Box-Behnken中心组合实验设计,采用响应面分析法分别优化复合保鲜液脱铝工艺与超声波辅助脱铝工艺。结果显示,未超声波辅助处理的优化条件：料液比1∶7（g/mL）、浸泡时间4 h、浸泡次数3次,铝残留量最低为126.4 mg/kg,浸泡时间影响最为显著;超声波辅助处理影响脱铝率大小的因素依次为超声时间 > 超声功率 > 超声频率 > 复合保鲜液浓度;其最佳工艺：超声频率28 kHz、超声功率272.6 W、复合保鲜液[V（醋酸）∶m（丙酸钙）=1∶2]浓度0.68%、脱除时间35 min,铝残留量为124.46 mg/kg,脱铝率达到85.3%;验证实验得到实际脱铝率为82.1%,与理论预测值相比,其相对误差约为3.2%。研究表明,与未超声波辅助相比,铝脱除率稳定,脱除时间由12 h缩短至1.75 h,工艺效率提高6.9倍。因此,超声波技术应用到海蜇产业将为高效开发低铝盐渍海蜇制品提供理论依据。     

超声波辅助提取泥鳅多糖的工艺优化

以泥鳅为原料,采用超声波辅助技术提取泥鳅多糖,运用硫酸-苯酚法检测泥鳅多糖的含量,利用单因素试验结果设计正交试验,探讨料液比、超声时间和超声功率对泥鳅多糖提取率的影响。经工艺优化,获得超声波提取泥鳅多糖的最佳工艺为料液比1∶20、超声时间20 min、超声功率100 W,在最佳工艺条件下提取泥鳅多糖提取率最高,达到2.79%。     

脱酸与脱腥条件对烤鳗油品质的影响

用正交试验法设计碱 (NaOH)加入量以优化烤鳗油的脱酸条件 ,并对脱酸后的酸价、过氧化值和收得率进行测定以评价其质量。运用减压水蒸气蒸馏法对烤鳗油进行脱腥处理 ,以气相色谱 -质谱 (GC -MS)联用技术测定经脱腥后烤鳗油的脂肪酸组成。结果显示 ,烤鳗油最佳脱酸条件为 0 .0 5 %过量碱 ,原碱质量分数 13.0 4 % ,温度6 0℃ ,处理时间 35min。经活性陶土脱色 ,可得到符合食用油标准的精炼烤鳗油 ,收得率为 80 % ;脱腥条件为温度180℃、压力 - 0 .0 9MPa ,水蒸气蒸馏处理 ,得到基本保持高不饱和脂肪酸组分的脱腥烤鳗油     

超声波酶解法提取鼠尾藻多糖

采用超声波及纤维素酶解法提取鼠尾藻多糖,探讨了功率、酶量、温度、时间对鼠尾藻多糖制备的影响,运用单因素设计正交试验,得出4种因素对鼠尾藻多糖提取的影响程度由大到小分别是时间、温度、超声波功率、酶量,优化得到最佳提取条件为超声波功率300 W、酶量3％、温度50℃、时间15 min；并通过对比分析,得出添加纤维素酶比对照组提取率提高37.03％.     

牡蛎清洗试验研究与清洗设备设计

分别采用毛刷、高压水、超声波对牡蛎进行清洗,以去除杂质比重为主要指标,探讨几种方式对牡蛎清洗效果的影响,初步确定最佳清洗参数。结果表明:毛刷、高压水、超声波都能够对牡蛎进行有效清洗。模拟人工冲刷式实验最佳工艺条件为:滚刷转速20 r/min,高压水压力8 MPa,流量15 L/min,时间2~5 min;超声波清洗实验最佳工艺条件为:低频(26 KHz),处理时间5 min。此系列工艺条件下牡蛎无死亡,生态冰温保活9 d后,存活率达到95%以上。以试验参数为基础,设计一种集三种清洗方式为一体的组合式牡蛎清洗设备。     

采用化学氧化法降解贝类养殖水中扑草净残留的研究

为去除贝类养殖过程中残留的农药扑草净,研究了不同处理(正常、超声波震荡、紫外线照射和不同pH)条件下高锰酸钾、过氧化氢和次氯酸钠对扑草净的降解效果。结果显示,次氯酸钠对扑草净的降解率最高,其降解效果不受紫外线照射、超声波震荡以及水体pH变化的影响。高锰酸钾和过氧化氢对扑草净的降解率较低,超声波震荡和紫外线照射辅助处理对降解效果没有影响,但当水体pH为10时,高锰酸钾和过氧化氢对扑草净的降解率显著提高。研究还对次氯酸钠降解扑草净的反应动力学进行了探讨,结果表明,扑草净的氧化降解符合二级反应动力学模型。     

超声波辅助酶法脱除石莼水溶性膳食纤维中蛋白质的工艺研究

本试验以蛋白质脱除率和水溶性膳食纤维(SDF)保留率作为指标,探讨各主要因素对石莼SDF脱蛋白效果的影响,并采用L_9(3~4)正交试验确定超声波辅助酶法脱蛋白的优化工艺参数。结果显示:在控制石莼SDF浓度0.6%和超声功率200 W的前提下,石莼SDF的超声波辅助酶法脱除蛋白优化工艺参数组合为:碱性蛋白酶使用量1 400 U/g、石莼SDF样液pH 11.0、超声温度50℃、超声时间1.5 h,以上述条件进行验证性试验,石莼SDF的蛋白质脱除率为(83.91±0.18)%,SDF保留率为(86.23±0.25)%,蛋白质脱除效果优于正交试验9个处理组。     

赤潮研究中圆海链藻实时荧光定量PCR检测方法的建立

为建立快速、准确的鉴定和定量检测赤潮生物的方法,以圆海链藻为例，以其18S rDNA序列为寻找种特异性引物的靶区域，通过分析18S rDNA序列，设计出适合用于RFQ-PCR的引物与探针，并通过引物PCR验证确定其特异性，进而以圆海链藻荧光定量PCR的引物和探针（Primer Thalassiosira rotula和Taqman Thalassiosira rotula），建立了定量检测圆海链藻的实时荧光定量PCR检测方法（RFQ-PCR）。与传统的显微镜计数方法比较，两者所获结果无显著性差异，证明了本方法的可行性，从而为我国沿海水域赤潮问题的研究提供了良好的技术检测途径。     

A reliable method for the induction of experimental furunculosis

Abstract. A series of experiments were carried out to assess the reproducibility of a bath challenge model to induce experimental furunculosis. The challenge method was shown to reliably induce a control mortality of 78.9 ± 3.8%. Although the method was shown to be robust, some tanks failed to achieve an acceptable control mortality. The reasons for this are unclear; however, the failure of a tank to induce a satisfactory control mortality is related to a rapid disappearance of the initial inoculum. The reason for the disappearance of the inoculum appears to be related to the presence of hydrophobic regions in the challenge tank.     

Cellulose acetate electrophoresis, radial immunodiffusion and nephelometry--comparative methods for the determination of albumin]

Different methods for the determination of albumin in dogs and cats are compared. Nephelometry was found to be the method of choice, due to its excellent precision, the immediate test results and the wide measuring range. The advantage of the radial immunodiffusion method is that expensive equipment need not be acquired, which makes the test profitable for small amounts of samples. At the expense of accuracy, electrophoresis enables the simultaneous evaluation of all protein fractions.     

鲁氏耶尔森菌qPCR快速检测方法的建立和应用

鲁氏耶尔森菌(Yersinia ruckeri)是导致虹鳟(Oncorhynchus mykiss)肠炎红嘴病的病原。本研究以鲁氏耶尔森菌毒力因子rup A基因为靶标设计1对特异性引物,以含有rup A基因部分序列的重组质粒为标准品构建标准曲线,优化建立检测鲁氏耶尔森菌的SYBR Green I实时荧光定量PCR检测方法,并对腹腔注射鲁氏耶尔森菌菌悬液的虹鳟肝、肾、脾、血液样品进行检测。结果显示,设计的引物具有良好的种间特异性,标准曲线线性方程为y=–3.3766x+40.012(R~2=0.9958);最低检测限为57 copy/μL,较常规PCR的灵敏度高出约100倍。应用检测结果表明,本方法可准确检测被鲁氏耶尔森菌感染的虹鳟样品。研究表明,所建立的qPCR方法具有特异、灵敏、快速、定量的优点,可用于快速诊断虹鳟肠炎红嘴病早期病症及定量检测鲁氏耶尔森菌。     

Validation of enzyme-linked immunosorbent assay for measurement of faecal cortisol in fish

Quantification of glucocorticoid (GC) levels in faeces has become an established method for the non-invasive assessment of adrenocortical activity. These hormones are frequently determined in plasma samples as parameters of adrenal activity and response to stress. Because GCs are metabolized and excreted with both intact hormone and their metabolites present in faeces, the concentration of GCs can be measured in excreta. Faecal samples present the advantages of easy collection, no stress to the animal and elimination of the issue of potentially misleading acute GC spikes. The aim of this study was to determine if an enzyme-linked immunosorbent assay (ELISA) for cortisol was appropriate for monitoring adrenocortical activity in faecal casts of fishes. Performance of the cortisol ELISA was validated by comparison to high-performance liquid chromatography, which is an established method for measuring free GCs and GC metabolites in faeces. Parallelism and sample extraction efficiency were compared for the two methods. Pearson's correlation across samples for these methods was 0.996. Results demonstrated that the ELISA was an efficient, sensitive and reliable method for cortisol measurement in faecal extracts, which should permit integration of non-invasive stress monitoring into studies of fish behaviour and physiology.     

A novel method of collecting fecal samples from spiny lobsters

A novel method is described for collecting feces from the spiny lobster Panulirus ornatus. Established methods for the collection of feces from crustacean species are not effective with P. ornatus. The feces produced by adults and juveniles of this species are not in a form that can be easily recovered by traditional methods. To overcome this problem, a plastic flange was glued onto the integument, around the anal pore. A small latex balloon was placed over the flange and provided a watertight collection vessel for voided feces. The flange was made from the modified cap from a 2 ml Eppendorf vial. Fixing the flange and changing the balloon after feeding was achieved without the use of anaesthetics and appears not to impose undue stress on the lobsters and they continued to feed. The method was successfully used to collect uncontaminated fecal samples from lobsters ranging from 150 to 800 g. This method may have wider application for other crustacean species.     

Improvement of the method for propagation,larval and postlarval rearing of the wels (Silurus glanis L.)

Using the old method of spawning it is impossible to produce wels larvae in quantities sufficient for a special fry farm economically. Because of this, a large-scale intensive method had to be developed, similar to those for other species which are propagated in hatcheries. A method for storing breeders was elaborated which included sewing their mouths shut and consequently eliminating injuries. Pituitary treatment — practised by others before — was applied, and disadvantages of the method (such as the development of furuncles) eliminated. A method for taking sperm from males with the help of a special syphon was developed.To avoid losses, larvae were reared in basins until they reached the size of 2–2.5 cm. Using this method considerable quantities of fry were produced in our hatchery.     

以栉孔扇贝闭壳肌为例的几种DNA提取方法的比较研究

以栉孔扇贝(Chlamys farreri)闭壳肌为试验材料，采用浓盐法、酚抽提法、LiCl法及改进的浓盐法和酚抽提法等提取其DNA，通过紫外吸收值测定及琼脂糖凝胶电泳法对提取的DNA质量进行分析比较。结果表明，这5种方法都能提到DNA，但改进的浓盐法和酚抽提法可将其中的蛋白质及RNA较为有效地除去，得到比较纯净的DNA。     

贝类派琴虫实时荧光定量PCR检测方法的建立和应用

选择派琴虫保守的核糖体DNAITS-2区域设计引物和TaqMan探针,通过对反应体系和反应条件进行优化,建立了实时荧光定量PCR检测派琴虫的方法。所构建方法检测质粒模板DNA的动态范围为2.6×10^1～2.6×10^7拷贝,敏感度可检测到26拷贝质粒DNA,而且与包拉米原虫、隐孢子虫等其他寄生性原虫无交叉反应,也不受贝类组织DNA的干扰。利用本研究所建立的方法对来自我国山东、福建等不同沿海海域的30份贝类样品进行检测,检出阳性样品3份。研究表明,本研究所构建的派琴虫实时荧光定量PCR检测方法具有快速、灵敏和特异等优点,可满足国内养殖场及进出口水生动物携带派琴虫的检测需要。     

Distribution of glutamine synthetase in the snapper (Pagrus auratus) and implications for the immune system

A sensitive radiotracer method was used to detect glutamine synthetase activity in fish. The distribution of glutamine synthetase activity using this method was shown to be widespread in organs and tissues of the marine finfish, snapper,Pagrus auratus. The optimum pH and temperature for glutamine synthetase activity and K_m of glutamate from the brain of the snapper are also reported. Interestingly no glutamine synthetase activity was detected in lymphoid organs, indicating, that as in the mammal, the teleostean immune system is likely to rely on glutamine in circulation for energy and nucleotide synthesis. Furthermore circulating glutamine levels fell as fish became stressed or became ill. Implications of these observations on the function of the immune system of fish are discussed.