地锦草对水产病原菌体外抗活性的研究

用95%的乙醇来提取地锦草(Euphorbia humifusa Wild)中的化合物,再分别用石油醚、乙酸乙酯、正丁醇和水对地锦草乙醇提取物进行分极性段萃取,研究地锦草各极性段二甲基亚砜萃取溶液对嗜水气单胞菌(Aeromonas hydrophila)、温和气单胞菌(Aeromonas sobria)、柱状屈挠杆菌(Flexibacter calumnaris)、鳗弧菌(Vibrisis angnillarumz)4种常见水产动物病原菌的抑菌效果。结果表明,地锦草各极性段的抑菌作用强度有明显差别(P<0.05),其中乙酸乙酯、石油醚极性段的抑菌作用最为强烈,对温和气单胞菌的MIC均≥12.5mg/mL;乙酸乙酯极性段对嗜水气单胞菌、鳗弧菌的MIC均≥50mg/mL;石油醚极性段对嗜水气单胞菌、鳗弧菌的MIC≥25mg/mL;正丁醇和水极性段对4种菌的MIC均≥50mg/mL。     

18种中草药煎液对温和气单胞菌的抑、杀菌效果

采用琼脂扩散法(打孔法)测定了五倍子等18种中草药对2株病原温和气单胞菌(Aerom onas sobria)的抑制作用,并采用试管2倍稀释法测定抑菌效果较强的中草药对这2株温和气单胞菌的抑菌和杀菌浓度。结果显示:五倍子、苏木、地锦草、石榴皮、黄芩、虎杖和山楂对病原温和气单胞菌的抑菌效果较强;苏木、五倍子、地锦草和石榴皮4种中药对2株温和气单胞菌的最小抑菌浓度(M IC)为1.56～6.25 mg/mL之间,最小杀菌浓度(MBC)为3.12～12.5 mg/mL之间,其中五倍子的抑菌和杀菌浓度最低,分别为1.56 mg/mL和3.12 mg/mL;而金银花等11种中草药抑菌作用较小。结果表明,所选取的18种中草药中五倍子对防治由温和气单胞菌感染引起的水产动物疾病最为有效。     

9种常见草药有效成份的提取及抑菌试验

用酒精回流法提取板兰根、大青叶、地锦草、斑地锦、乌蔹莓、蒲公英、马齿苋、车前草、铁苋菜等9种常见中草药的有效成份，并分别用强毒株嗜水气单胞菌、温和气单胞菌进行体外抑菌，抑菌效果由大到小分别为地锦草＞马齿苋＞斑地锦＞板兰根＞铁苋菜＞蒲公英＞大青叶＞车前草＞乌蔹莓；临床使用地锦草、马齿苋、板兰根合剂60g/m^3及痢特灵1.5g/m^3治疗由鲁氏不动杆菌与温和气单胞菌引起的欧洲鳗鲡红头病取得明显效果。     

16种中草药提取物对嗜水气单胞菌的体外抑菌实验

测定了16种中草药对嗜水气单胞菌(Aeromonas hydrophila)的体外抑菌作用,筛选出对嗜水气单胞菌有较好体外抑制作用的中草药.用煎煮法和乙醇蒸馏法2种方法提取中草药有效成分,采用琼脂平板扩散法研究了黄连、五倍子、大黄、诃子等16种中草药对嗜水气单胞菌的体外抑菌作用,测定了抑菌圈直径.实验结果表明,16种中草药提取物对嗜水气单胞菌有不同程度的抑制作用,五倍子、黄连、诃子的水提物和醇提物的抑菌效果较好,抑菌圈平均直径均在15 mm以上,但中草药的水提物和醇提物的抑菌效果存在不同程度的差异.     

细菌性"颤抖病"病原的体外中药药敏试验

嗜水气单胞菌2菌株ES-1、ES-2对地锦草、黄莲、五倍子和增效剂具有高度敏感性，这4种药物可作为防治由该菌引起疾病的首选药物，同时发现这2株菌对黄芪、穿心莲、连翘等具有不同程度的耐药性。通过3种高度抑菌药物间及增效剂之间的相互作用，得出增效剂对五倍子、黄莲有明显的增效作用，地锦草和五倍子具有协同作用，尤其增效剂对五倍子的增效作用效果最为明显，该复方药物可作为由该菌引起疾病的高效、低成本、低毒的防治药物。     

中草药复方B总黄酮的提取及其抑菌效果

对中草药复方B总黄酮的提取及其对败血病痛原菌-嗜水气单胞菌的抑菌效果进行了研究。提取试验的结果：用30倍量的85％的乙醇76℃条件下回流提取3次，每次2小时为最佳方案；抑菌试验的结果：复方B的提取物总黄酮抑菌效果较好。     

水产动物气单胞菌鉴定方法研究进展

<正>气单胞菌属于γ-变形菌纲、气单胞菌目、气单胞菌科、气单胞菌属(Aeromonas),该属包括嗜水气单胞菌(A.hydrophila)、温和气单胞菌(A.sobria)、豚鼠气单胞菌(A.caviae)、维氏气单胞菌(A.veronii)和杀鲑气单胞菌(A.salmonicida)等20个种,其中嗜水气单胞菌和杀鲑气单胞菌又分别包括5个亚种[1]。气单胞菌广泛分布于自然生态系统     

被动转移免疫在鲫感染温和气单胞菌中的作用

实验用鲫平均体重100g，采用免疫物质被动转移试验，探讨体液免疫在鲫感染温和气单胞菌（Aeromonas sobria)中的保护作用。结果表明，无论免疫物质是来自异源还是自源，是由相关的嗜水气单胞菌（A.hydrophia)诱导产生还是由攻毒的温和气单胞直接刺激，均具有良好的免疫保护作用。实验还发现，再次攻毒时，在所有抗体组的鲫存活率为100%时，全血被动转移组的死亡率高达55.6%；免疫前后免疫复合物发生明显变化。说明抗体在保护鲫免疫受温和气单胞菌的致死性感染中发挥决定性的作用。     

中华鳖气单胞菌菌细胞苗和菌化学成份疫苗研制

利用分离自中华鳖的嗜水气单胞菌Ａｈ９６１００４、Ａｈ９６０９１６和Ａｈ９７０５１６与温和气单胞菌Ａｓ９７０５１０菌体及其上清制备灭活苗细胞苗和化学成份疫苗。肌肉注射中华鳖进行免疫，加强免疫１５ｄ后，用菌株Ａｈ９６１００４腹腔注射进行攻击，观察２０ｄ。结果表明，温和气单胞菌Ａｓ９７０５１０株的菌苗及其胞外分泌物（ＥＣＰ）苗组和嗜水气单胞菌Ａｈ９６１００４株的外膜成份组对嗜水气单胞菌的攻击的保护率最高     

鳖三种气单胞菌的分离与药物敏感试验

在1997～2001年对广东、湖南等地70个鳖养殖场进行常见病的调查研究、分离纯化病原后进行生化试验，鉴定出11株嗜水气单胞菌、11株豚鼠气单胞菌、6株寡源气单胞菌等，经过毒力试验后．选出较强毒力的11株细菌，用15种药物进行敏感性试验筛选出敏感药物，为药物的临床应用提供依据。     

微波辅助提取石榴皮中抗氧化成份研究

本试旨在研究提取石榴中抗氧化成份。以石榴皮作为原料,95%的乙醇作为溶剂,采用微波辅助提取石榴皮中的抗氧化成分,以产率为评价指标,考察了微波功率(160W、320W、480W、640W)、微波处理时间(40s、60s、80s、100s)和料液比(1:5、1:7、1:9、1:11)对产率的影响;在单因素考察基础上,进行正交实验设计:微波功率(320W、480W、640W)、微波处理时间(40s、60s、80s)和料液比(1:7、1:9、1:11),结果表明:乙醇浓度95%、微波功率640W、微波处理时间60s、料液比1:11为最优工艺条件。结果提示,用微波辅助提取石榴皮中抗氧化活性物质是较好的方法。     

Acute LD50 and kidney histopathology following injection of erythromycin (Erythro-200) and its carrier in spring chinook salmon, Oncorhynchus tshawytscha (Walbaum)

We evaluated the toxicity of commercial injectable erythromycin (Erythro-200) and the injectable drug carrier alone (PEG-400, ethyl alcohol and ethyl acetate) in sub-adult spring chinook salmon, Oncorhynchus tshawytscha (Walbaum). Both compounds were toxic to salmon when administered in high dosages. The 50% lethal dose (LD₅₀) of the erythromycin within its carrier (Erythro-200) at 12 °C was estimated at 429 mg erythromycin kg^–1 or 2.145 mL kg^–1 when measured by volume. The LD₅₀ of the drug carrier alone was 2.95 mL kg^–1 by volume or equivalent to 3.21 g kg^–1 of the carrier. We evaluated and scored the histopathology in kidney sections removed from fish 96 h after injection with Erythro-200, the drug carrier alone, or sterile phosphate buffered saline (PBS) that served as a reference control. Kidneys from fish injected with saline had lowest average pathology scores; those injected with the carrier had moderate scores; and the pathology score was highest in tissues from erythromycin-injected fish. In tests at 12 °C, vacuoles were more prominent in proximal and distal tubules of fish injected with higher dosages of 403 and 545 mg erythromycin kg^–1 than in tissues from fish injected with 299 mg kg^–1.     

中华绒螯蟹颤抖病药物治疗的初步研究

用水产常用清塘剂、杀菌驱虫药、消毒剂、中草药及自配药物克螺威1号(SI-1)对中华绒螯蟹颤抖病病原螺原体进行体外药敏试验。结果表明,螺原体对生石灰、茶籽饼、敌杀死、硫酸铜、硫酸锌、次氯酸钙、三氯异氰脲酸、二氧化氯、溴氯海因、二溴海因、高锰酸钾、五倍子、黄连、黄柏及自配药物克螺威1号具有不同程度的敏感性。通过最小杀菌浓度(MBC)比较,螺原体对清塘剂茶籽饼、杀菌驱虫药硫酸铜、消毒剂次氯酸钙、中草药黄连、自配药物克螺威1号更为敏感,其中克螺威1号为最敏感药物,最小抑菌质量浓度(M IC)为0.16 mg/L,MBC为2.5 mg/L。对克螺威1号进行急性毒性试验和颤抖病治疗试验表明,克螺威1号的96 h LD50为1106.62 mg/kg,安全使用量为110.66 mg/kg。用其治疗人工诱发颤抖病的中华绒螯蟹,克螺威1号具有显著效果(P<0.05),随后在10个被确诊为螺原体引发中华绒螯蟹颤抖病的养殖场进行药物治疗试验,在6个养殖场治疗有效,而在其他4个养殖场治疗无效,治疗试验效果不一,用克螺威1号治疗中华绒螯蟹颤抖病具有较好的前景,但仍需进一步做大面积长期临床试验。     

Supercritical CO<Subscript>2</Subscript> extraction and concentration of n−3 polyunsaturated fatty acid ethyl esters from tuna cooking juice

Cooking is an important step in the tuna canning process; it usually produces functional solubilized proteins and lipids. The objective of this study was to recover and increase the concentration of functional n−3 fatty acids from the lipids in cooking juice. The lipids contained 12.98% eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) after ethyl esterification and increased to 37.4% with subsequent urea adduct fractionation. Supercritical carbon dioxide extraction was further used to increase the concentration of EPA and DHA ethyl esters. The ratio of EPA+DHA ethyl esters (high-molecular-weight components) to that of C₁₆ and C_18∶1 fatty acid ethyl esters (low-molecular-weight components) was used as a separation index for evaluating the process variables. Experimental results indicated that a high CO₂ density caused a low separation factor. At 1500 psig and 328.2 K, the extraction collected over 600 L CO₂, displaying an accepted concentration factor of EPA+DHA ethyl esters herein. About 80% yield of EPA and DHA was obtained and the ethyl esters increased from 37.4% to 54.3%.     

n_3多价不饱和脂肪酸营养强化轮虫技术研究

通过甲酯型，乙酯型，脂肪酸型和甘油酯型化学型式ｎ－３多价不饱和脂肪酸的营养强化饵料对轮虫的营养强化试验，观察了营养强化过程中轮虫生理生态变化，测定了轮虫体内ＥＰＡ，ＤＨＡ的含量。结果表明：不论是哪种化学型式的营养强化饵料，营养强强效果均好于微绿球藻强经对照组，轮虫中ＥＰＡ，ＤＨＡ含量明显提高，并在强化１２ｈ达到最高峰。     

4株细菌代谢产物增强鲫鱼免疫功能有效活性部位的研究

分别以蕈状芽孢杆菌(Bacillus mycoides)、皮杆菌(Dermabactersp)、噬菌蛭弧菌(Bdell ovibro bacteriovorus)和蜂房芽孢杆菌(Bacillus alvei)4株细菌代谢产物不同溶剂(石油醚、三氯甲烷、乙酸乙酯、正丁醇)的提取物腹腔注射鲫鱼(Carassius auratus),通过测定注射提取物后鲫鱼的血清凝集抗体效价、离体白细胞吞噬活性、吞噬细胞杀菌活性,研究不同代谢产物、不同提取物对鲫鱼免疫功能的影响,以确定代谢产物中活性物质的有效活性部位。结果表明:蕈状芽孢杆菌代谢产物中的活性物质主要集中在石油谜和乙酸乙酯提取物中;噬菌蛭弧菌代谢产物中的活性物质主要集中在石油醚和三氯甲烷提取物中;蜂房芽孢杆菌代谢产物中的活性物质主要集中在石油醚提取物中;而皮杆菌代谢产物中的活性物质则集中在乙酸乙酯和石油醚提取物中。试验结果为进一步追踪分离活性单体提供了基础资料。     

水产品中氯霉素、甲砜霉素和氟甲砜霉素残留量高效液相色谱-串联质谱内标测定方法的研究

建立了水产品中3种氯霉素类抗生素残留的高效液相色谱-串联质谱(HPLC-MS/MS )测定法.以氘代氯霉素(d5-CAP)为内标,样品在碱性条件下用乙酸乙酯提取,提取液氮气吹干,正己烷液液分配脱脂后,采用配有ESI源的LC-MS/MS选择反应监测(SRM)负离子模式测定,可同时对水产品中的氯霉素、甲砜霉素和氟甲砜霉素进行定性和定量测定.方法简化了样品前处理过程,省去固相萃取步骤,具有操作简便、有机试剂消耗量少、测定周期短和定量更准确等优点.方法的检出限:氯霉素为0.01 μg/kg,甲砜霉素和氟甲砜霉素为0.03 μg/kg.     

Retention efficiency and release of nutrients in the digestive tract of larval shrimp (Penaeus japonicus Bate) using different microencapsulated diets

Retention efficiency and release of the nutrients in the digestive tract of larval shrimp (Penaeus japonicus) using the diets microencapsulated with gelatin and ethyl cellulose respectively were evaluated. The microencapsulated diets were produced using the fluidized bed coating process. 7.8 % gelatin and 4.2 % ethyl cellulose to the whole microencapsulated diet were respectively adopted as coating material. After immersion in 35 ‰ NaCl solution for 1 h, the nitrogen retention efficiency was significantly lower in the diet microencapsulated with gelatin compared with the diet microencapsulated with ethyl cellulose. Free amino acid retention efficiency of the diet microencapsulated with gelatin and ethyl cellulose was 12.9, 17.2 %, respectively. The mysis II larval shrimp (P. japonicus) 10 days after hatch were fed two different microencapsulated diets for 20 days and grew significantly more than the control larval shrimp fed with Artemia and shrimp flake. The nutrient components in intermediate intestine of larval shrimp were increased gradually in the order of the control (50 % shrimp flake + 50 % Artemia), Group I (50 % diet microencapsulated with gelatin + 25 % shrimp flake +25 % Artemia), Group II (100 % diet microencapsulated with gelatin), and Group III (100 % diet microencapsulated with ethyl cellulose). This study confirmed that the microencapsulated diets with slow and controlled release characteristic in the digestive tract of the larval shrimp (P. japonicus).     

Evaluation of ovum storage techniques for reproductive studies of fishes

Abstract– We compared dry masses of mature oocytes and ripe eggs from Etheostoma lynceum and Etheostoma caeruleum that were stored in varying concentrations of either formalin or isopropyl alcohol. In comparison to 10% formalin, alcohol significantly reduced the weights of both mature oocytes and ripe eggs, with 25% isopropyl alcohol resulting in much smaller reductions than 50% isopropanol. Three percent formalin resulted in smaller decreases in the masses of oocytes and eggs than the alcohol treatments, and not all comparisons with 10% formalin were significantly different. We recommend fixing specimens and storing them and gametic cells removed from them in 10% formalin. In cases where specimens or eggs in alcohol must be used in addition to material in formalin, correction factors (to formalin standard) should be developed, but the correction should not be made uncritically due to the varying effects alcohol may have among samples.