双齿围沙蚕化学成分及其浸膏抗肿瘤活性的研究

采用硅胶柱色谱及薄层制备色谱等手段从双齿围沙蚕(Perinereis aibuhitensis)中分离纯化出2种化合物。对所获得的化合物通过理化常数测定、波谱数据分析同文献对照的方法进行了结构鉴定。同时,采用CCK-8方法对双齿围沙蚕浸膏及乙酸乙酯萃取部分分别进行抑制肿瘤细胞活性测试。结果表明,从双齿围沙蚕中分离得到2种化合物分别为:胆甾-4烯3α,6β醇(1)和(E)-17-(4,7-dimethyloct-5-en-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(2),且化合物1是首次从沙蚕中分离得到。抑制肿瘤细胞活性测试表明,70%乙醇浸提获得的双齿围沙蚕浸膏及乙酸乙酯萃取部分对人胃癌细胞AGS、人肝癌细胞HepG2以及人卵巢癌细胞SKOV3均具有抑制细胞增殖作用,且具有剂量和时间依赖性,可作为潜在的抗肿瘤药物进行进一步研究。     

海胆壳中具有抑菌活性物质的分离鉴定

观察海胆壳中化学成分对金黄色葡萄球菌的抑制活性,对其进行分离纯化,并根据理化性质、波谱分析和文献对照的方法对所分离的化合物进行结构鉴定。用75%EtOH对粉碎的海胆壳进行热浸提,减压浓缩后依次用石油醚、乙酸乙酯和正丁醇进行萃取,得到各层提取物,对其进行抑制金黄色葡萄球菌生物活性测定。进而,粗提物利用薄层层析、反复硅胶柱层析、半制备薄层色谱层析以及重结晶等手段进行分离纯化得到化合物。通过抑制金黄色葡萄球菌生物活性测定可知石油醚提取物、正丁醇提取物均有较显著的抑制金黄色葡萄球菌的活性。石油醚提取物经过反复硅胶柱层析和重结晶手段得到化合物HDB,结构鉴定为棕榈酸。正丁醇提取物经过反复硅胶柱层析、半制备薄层色谱以及重结晶等手段分离得到化合物HDA,结构鉴定其为(3β)-胆甾-5-烯-3-醇。     

海绵共附生放线菌Streptomyces parvulus MA1076的次级代谢产物研究

海绵共附生放线菌是活性次级代谢产物的重要来源,对海绵共附生放线菌Streptomyces parvulus MA1076进行菌株鉴定和化学成分分析。通过16S rDNA序列分析、构建系统发育树进行菌株鉴定;利用硅胶柱色谱、Sephadex LH-20凝胶柱色谱以及半制备高效液相色谱等技术对其发酵产物进行分离和纯化,通过核磁共振、高分辨质谱等方法,并结合文献数据对化合物进行结构鉴定。从放线菌Streptomyces parvulus MA1076中分离鉴定出6个化合物,分别为放线菌素D(化合物1)、胡萝卜苷(化合物2)、环-((S)-脯氨酸-(R)-亮氨酸)(化合物3)、胸腺嘧啶核苷(化合物4)、3′-氧-乙酰胸腺嘧啶核苷(化合物5)和3-羧基吲哚(化合物6)。化合物2～6为首次从该种属放线菌中分离得到。活性筛选结果显示,化合物1对两株胰腺癌细胞系有明显的增殖抑制活性,研究结果丰富了海绵共附生放线菌次级代谢产物的结构多样性,可为开发利用海洋微生物资源提供参考。     

香樟和银杏落叶海水浸提液对东海原甲藻的抑藻效应及其有效成分分析

为寻找防治赤潮的有效手段,以香樟(Cinnamomum camphora)和银杏(Ginkgo biloba)落叶作为抑藻制剂材料,探究其常温海水浸提液对东海赤潮高发区原因种东海原甲藻(Prorocentrum donghaiense)的化感抑制效果,并分别采用顶空-固相微萃取-气相色谱/质谱联用法和超高效液相色谱串联四极杆-飞行时间质谱法分析了香樟和银杏海水浸提液中的有效成分。结果表明：香樟和银杏叶浸提液对东海原甲藻生长的96 h半效应浓度(96 h-EC₅₀)分别为1.23 g·L^-1和1.22 g·L^-1。高浓度香樟叶浸提液(≥3.0 g·L^-1)处理72 h时即可使大量东海原甲藻细胞破裂,96 h时对东海原甲藻生长的抑制率高于70%,且在第8天仍可抑制东海原甲藻的生长。银杏叶浸提液对东海原甲藻的抑制作用起效快,处理24 h时,高浓度试验组(≥3.0 g·L^-1)中东海原甲藻基本失去游动能力,处理96 h时,浓度≥2.0 g·L^-1的银杏叶浸提液对东...     

2株海鞘共附生真菌的次生代谢产物研究

分别对采集自防城港白龙的皱瘤海鞘(Styela plicata)和北海南汅的冠瘤海鞘(Styela canopus)样品的共附生真菌Purpureocillium sp. FBZ-1和Penicillium sp. BNG-1进行研究。通过显微镜观察鉴定其形态,并通过分析生物学特征鉴定其种属。通过对两种真菌的次级代谢产物进行分离和结构鉴定,共分离鉴定28个小分子化合物,从中未发现新的化合物,主要的化合物类型是芳香胺类、生物碱类和二酮哌嗪类,分别为:lumichrome、 1,2,3,4-四氢-β-咔啉-3-羧酸、β-咔啉、环(异亮氨酸-脯氨酸)、环(亮氨酸-羟基脯氨酸)、环(色氨酸-缬氨酸)、环(亮氨酸-脯氨酸)、环(苯丙氨酸-脯氨酸)、环(异亮氨酸-丙氨酸)、环(色氨酸-亮氨酸)、苯乙胺、N-乙酰苯乙胺、吲哚乙胺、N-甲基色胺、N-(4-羟基)丁酰苯乙胺、对羟基苯乙胺、 5,7,4-三羟基异黄酮、 7,4-二羟基异黄酮、 5,7,3,4-三羟基异黄酮、邻苯二甲酸异辛酯、 2,4-N,N-dimethyl-lumichrome、 3,4-二氢-3-甲基-β-咔啉-1-酮、色氨酸、 2-羟基-3-吲哚丙酸、环(缬氨酸-脯氨酸)、环(亮氨酸-丙氨酸)、环(苯丙氨酸-5-甲基-3,4-脱氢脯氨酸)、环(色氨酸-天冬酰胺)。研究结果对利用我国丰富的海鞘及其共附生微生物资源并研究其次级代谢产物结构多样性,发现其中可能含有的结构新颖的化学物质以开发抗菌、抗病毒及抗肿瘤新药,具有十分重要的理论意义和潜在的应用价值。     

星座短腹海鞘脂溶性成分的提取工艺

以星座短腹海鞘(Aplidium constellatum)为研究对象,采用正交试验法对其脂溶性成分的提取工艺进行优化。以提取率和类胡萝卜素含量为指标评定工艺的优劣,并通过气相色谱对提取物中二十碳五烯酸(EPA)与二十二碳六烯酸(DHA)进行相对含量分析。结果表明,采用料液比1∶1.5(W/V)、提取时间8 min、溶剂(正丁醇/乙酸乙酯)配比1∶2(V/V)工艺条件最佳。正交试验提取物的总不饱和脂肪酸的相对含量达63.40%~65.00%,其中EPA和DHA总量在15.20%~16.30%。综合2个指标,正交试验法优化星座短腹海鞘脂溶性成分的提取工艺是可行的;利用该提取方法获得的星座短腹海鞘脂溶性提取物中,EPA和DHA两种ω-3多不饱和脂肪酸的相对含量较高,具有很好的开发前景。     

韩国的真海鞘养殖

真海鞘　(HalocynthiaroretziDrasche)属于脊索动物门、尾索亚门、海鞘纲[1] 。多分布于日本太平洋侧的三陆沿海和日本海男鹿半岛以北海域。在韩国多分布于东海岸和南海岸。真海鞘营养丰富、味道鲜美,尤其夏季味道最鲜美,深受日韩国民的青睐,以生食为主,同时远销日本、意大利、法国等国家。2 0世纪6 0年代,韩国真海鞘的自然资源量很多,70年代中期,由于不明原因的大量死亡[2 ] ,资源量急剧下降,濒临灭绝。为了恢复真海鞘资源,韩国从日本引进苗种开始了垂下延绳式养殖生产。进入80年代,养殖产量趋向稳定,现已形成产业化规模,自然资源也有一定…     

大洋纵列海鞘共附生链霉菌Streptomyces sp.FDZ35-2的化学成分

海鞘共附生微生物是尚待关注的海洋微生物资源重要来源之一。本文研究了大洋纵列海鞘(Symplegma oceania)的一株共附生链霉菌Streptomyces sp. FDZ35-2的代谢产物,以探讨其主要代谢产物的结构类型和菌株的潜在药用价值。运用多种柱层析和HPLC等分离技术,从菌株甲醇提取物中分离鉴定2个主要的单体化合物;经LC-MS、1H-和13C-NMR等波谱学方法和文献数据对照,鉴定化合物结构分别为:9-甲基-β-咔啉-3-羧酸甲酯(1)和放线菌素D(2)。考虑放线菌素D生物合成可能利用培养基氮源中的氨基酸,本文还利用磷虾蛋白水解产物代替商品蛋白胨作为培养基氮源,对Streptomyces sp. FDZ35-2的发酵条件进行了初步研究,结果显示能产出更多种放线菌素D的结构类似物的发酵培养基为:磷虾蛋白水解产物2.0 g、葡萄糖2.0 g、硫酸铵1.0 g、碳酸钙0.5 g、氯化钠0.05 g、磷酸二氢钾0.05 g、海水1.0 L,p H 7.8~8.0。对比实验结果表明:与商品蛋白胨组相比,南极磷虾蛋白水解产物作为发酵培养基氮源所发酵产生的放线菌素D的结构类似物多样性更好,二者产生总放线菌素含量相当。这为开发南极磷虾蛋白在微生物发酵工业中的新用途提供了依据。     

南极曲霉Aspergillus niger NJ49次级代谢产物的分离与鉴定

为挖掘南极真菌的资源应用潜力，对一株南极真菌进行了分子鉴定、次级代谢产物分离及结构鉴定。经ITS rDNA序列分析并构建系统发育进化树，鉴定该菌株为曲霉属真菌，命名为Aspergillus niger NJ49。综合运用Sephadex LH-20和G25凝胶柱层析、半制备高效液相色谱等方法对南极磷虾共附生微生物曲霉属真菌A.niger NJ49的发酵物进行分离和纯化，并利用核磁、质谱等现代波谱学方法进行结构鉴定。从南极曲霉A.niger NJ49中分离鉴定出12个化合物，分别鉴定为Trp-Val-Val(1)、Phe-Val-Val(2)、cyclo-(His, Leu)(3)、cyclo-(Pro, Arg)(4)、(3S,12aS)-3-(Isopropyl)-2,3,6,7,12,12a-hexahydro-pyrazino[1′,2′:1,6]-pyrido[3,4-b]-indole-1,4-dione(5)、cyclo-(Pro, Phe)(6)、cyclo-(Val, Phe)(7)、cyclo-(Pro, Tyr)(8)、cyclo-(Pro, Val)(9)、Phe...     

青海湖裸鲤特征挥发性成分分析

为分析青海湖裸鲤(Gymnocypris przewalskii)特征挥发性成分,以体重为20 g的青海湖裸鲤作为研究对象,剖取肌肉后采用固相微萃取(SPME)结合气相色谱-质谱联用(GC-MS)对裸鲤肌肉挥发性成分进行分离鉴定。结果表明,在青海湖裸鲤肌肉中检测出60种化合物,包括8种醇类化合物、9种醛类化合物、5种酮类化合物、2种酸类化合物、8种酯类化合物、12种烃类化合物、12种芳香族化合物及4种其他类物质,其中芳香族化合物含量最高。结合气味活度值(OVA)分析,青海湖裸鲤肌肉挥发性气味的主要贡献化合物为1-辛烯-3-醇、己醛、辛醛、壬醛、2-癸烯醛及2,4-癸二烯醛。     

Morphological characterization of the tunic in the edible ascidian, Halocynthia roretzi (Drasche), with remarks on 'soft tunic syndrome' in aquaculture

'Soft tunic syndrome' is a serious problem in the aquaculture of the edible ascidian, Halocynthia roretzi (Drasche), and often leads to mass mortality. Here, we describe the tunic morphology of intact and diseased ascidians to reveal structural differences between them. Morphologically, diseased tunics are not very different from intact tunics, although the former are thinner and softer than the latter. While several types of cells are distributed in the tunic, the cell types and their cytomorphologies were almost identical in both groups. As bacterial/protozoan cells were not found in either intact or diseased tunics, they are not the direct cause of soft tunic syndrome. The most remarkable difference was in the bundles of tunic fibres that compose the tunic matrix; in intact tunics, the thick bundles interlace to form a firm matrix, whereas in soft tunics, the tunic fibres do not form thick bundles. Furthermore, areas of low fibre density were found in diseased tunics. Therefore, soft tunic syndrome probably causes inhibition of bundle formation and degradation of tunic bundles, creating areas of low fibre density, although the causes remain unknown.     

Tunic morphology and viral surveillance in diseased Korean ascidians: soft tunic syndrome in the edible ascidian,Halocynthia roretzi (Drasche), in aquaculture

‘Soft tunic syndrome’ causes mass mortality in the edible ascidian Halocynthia roretzi in Korean and Japanese aquaculture. In histopathological comparison, there were no specific differences between diseased specimens from Korea and Japan, indicating that soft tunic syndrome occurring in Korea and Japan is the same disease. No bacterial or protozoan cells were microscopically detected in either healthy or diseased tunics suggesting they are not the direct causes of soft tunic syndrome. Attempts were made to isolate virus from affected ascidians taking into account temperature conditions in which soft tunic syndrome is most prevalent in the field. However, no viruses were isolated from diseased or non‐diseased specimens using chinook salmon embryo (CHSE‐214), flounder fin (FFN) or epithelioma papillosum cyprini (EPC) cell lines.     

In vitro and in vivo efficacy of drugs against the protozoan parasite Azumiobodo hoyamushi that causes soft tunic syndrome in the edible ascidian Halocynthia roretzi (Drasche)

It was discovered recently that infection by a protozoan parasite, Azumiobodo hoyamushi, is the most probable cause for soft tunic syndrome in an edible ascidian, Halocynthia roretzi (Drasche). In an attempt to develop measures to eradicate the causative parasite, various drugs were tested for efficacy in vitro and in vivo. Of the 20 antiprotozoal drugs having different action mechanisms, five were found potent (24‐h EC₅₀ < 10 mg L^?1) in their parasite‐killing effects: formalin, H₂O₂, bithionol, ClO₂ and bronopol. Moderately potent drugs (10 < 24‐h EC₅₀ < 100 mg L^?1) were quinine, fumagillin, amphotericin B, ketoconazole, povidone‐iodine, chloramine‐T and benzalkonium chloride. Seven compounds, metronidazole, albendazole, paromomycin, nalidixic acid, sulfamonomethoxine, KMnO₄, potassium monopersulphate and citric acid, exhibited EC₅₀ > 100 mg L^?1. When ascidians were artificially infected with A. hoyamushi, treated using 40 mg L^?1 formalin, bronopol, ClO₂, or H₂O₂ for 1 h and then monitored for 24 h, very low mortality was observed. However, the number of surviving parasite cells in the ascidian tunic tissues was significantly reduced by treating with 40 mg L^?1 formalin or ClO₂ for 1 h. The data suggest that we might be able to develop a disinfection measure using a treatment regimen involving commonly available drugs.     

Growth performance and the soft body composition of juvenile abalone,Haliotis discus,Reeve 1846, fed the extruded pellets substituting fish meal and macroalgae with tunic meal of sea squirt,Halocynthia roretzi

Growth performance and the soft body composition of juvenile abalone fed the extruded pellets (EPs) substituting fish meal (FM) and macroalgae (MA) with tunic meal of sea squirt (SS) was investigated. A total of 1,260 abalone were distributed into 18 containers. Six experimental diets were prepared in triplicate. Five diets were pelletized by an extruder pelleter. The 140 g/kg FM and 250 g/kg mixture of MA were included into the control (Con) diet. Five hundred and 1,000 g/kg of each FM and MA were substituted with an equal amount of tunic meal of SS, referred to as the FM50, FM0, MA50 and MA0 diets, respectively. Finally, dry Undaria pinnatifida was prepared. Weight gain and specific growth rate of abalone fed all EPs were greater than those fed U. pinnatifida. Weight gain of abalone fed MA50 and FM50 diets was greater than Con and FM0 diets, but not different from MA0 diet. Higher crude protein and lipid contents were observed in soft body of abalone fed all EPs compared to U. pinnatifida. In conclusion, FM and MA up to 500 and 1,000 g/kg, respectively, could be replaced with tunic meal of SS in EPs without retardation in growth of abalone.     

Substitution effect of sea tangle (ST) (Laminaria japonica) with tunic of sea squirt (SS) (Halocynthia roretzi) in diet on growth and carcass composition of juvenile abalone (Haliotis discus,Reeve 1846)

Substitution effect of sea tangle (ST) with tunic of sea squirt (SS) in diet on growth and carcass composition of juvenile abalone was determined. One thousand four hundred and seventy abalones were distributed into 21 containers. Six formulated diets in triplicate were prepared. A 200 g/kg ST was included into the ST0 diet. The 200, 400, 600, 800 and 1000 g/kg of ST were substituted with the same amount of tunic of SS, referred to as the ST200, ST400, ST600, ST800 and ST1000 diets, respectively. Finally, Undaria was prepared to compare effect of the formulated diets on performance of abalone. The experimental diets were fed to abalone for 16 weeks. Weight gain of abalone fed the ST400 diet was higher than that of abalone fed the ST0, ST600, ST800 and ST1000 diets and Undaria. Weight gain of abalone fed the formulated diets was higher than that of abalone fed the Undaria. The chemical composition of the carcass of abalone was affected by dietary substitution of ST with tunic of SS. In conclusion, ST could be completely substituted with tunic of SS without retardation in performance of abalone. Abalone fed the ST400 diet substituting 400 g/kg ST with tunic of SS achieved the best growth.     

水硬度对七彩神仙鱼幼鱼发育的影响

采用不同硬度的水对七彩神仙鱼幼鱼进行饲养。6周龄幼鱼在硬度为7.94°dH±0.30°dH时饲养84d后,比在硬度为14.71°dH±0.23°dH水中的幼鱼个体大,生长速度快。表明较高硬度的水有利于七彩神仙鱼幼鱼的生长发育。     

虎斑乌贼受精卵卵黄营养成分分析

本实验对虎斑乌贼受精卵卵黄的营养成分进行分析,旨在探讨其幼体的营养需求量,为其幼体配合饲料研制提供参考数据。随机选取大约800个虎斑乌贼受精卵的卵黄,采用国家标准方法测定其水分、灰分、粗蛋白质、粗脂肪、氨基酸、脂肪酸和矿物元素含量。结果表明:1)虎斑乌贼受精卵卵黄中粗蛋白质含量为76.33%(干重基础);总氨基酸(TAA)和必需氨基酸(EAA)含量分别为71.22%和32.38%(干重基础),EAA/TAA为45.46%,氨基酸中以谷氨酸(Glu)含量最高(9.97%),必需氨基酸中亮氨酸(Leu)含量最高(7.58%)。2)其粗脂肪含量12.71%(干重基础);共检出17种脂肪酸,包括8种饱和脂肪酸(SFA)、5种单不饱和脂肪酸(MUFA)和4种多不饱和脂肪酸(PUFA),SFA、MUFA和PUFA分别占脂肪酸总量的43.47%、7.54%和49.25%,其中以DHA含量最高,达32.80%,EPA含量为7.70%,DHA/EPA为4.26。3)检测出Na、K、Ca、Mg、Sr、Mn、Fe、Cu、Zn、Al和As 矿物元素,微量元素中富含Zn、Al和Fe,含量分别为 0.77、0.71和0.43 mg/kg(鲜重基础)。由此可见,卵黄具有高蛋白、低脂肪,富含n-3PUFA的特点;虎斑乌贼幼体饲料中蛋白质需求量参考值为76.33%;氨基酸需求量参考值,如赖氨酸(Lys)为5.49%,蛋氨酸(Met)为2.63%;脂肪的需求量参考值为12.71%,DHA为4.17%,EPA为0.98%;微量元素需求量参考值,如Zn为2.77 mg/kg,Cu为0.19 mg/kg(干重基础)。     

斑节对虾虾苗活力检测方法

该研究通过肉眼观察、镜检，进行干露、饥饿、盐度突降、福尔马林等抗性试验，并采用病毒检测等方法，以期建立评估斑节对虾（Penaeus monodon）虾苗活力和质量标准。结果表明，斑节对虾健康虾苗具有趋光性、集群性，体表光洁，肌肉透亮，肠胃食物充盈等特性。测试虾苗干露时间以15min为宜，健康虾苗干露后能立即恢复活力，而病弱虾苗多出现死亡、昏迷现象；虾苗的成活率随饥饿时间的延长而降低，随福尔马林浓度升高和时间延长而降低，随盐度突降幅度增加而降低。健康虾苗能忍受100～200μL·L^-1福尔马林溶液30min，成活率近100％；在盐度20～30下虾苗的成活情况较好，而其在淡水中仅能存活1h。对虾苗进行病毒检测，可以避免养殖中因虾苗携带病毒而可能导致的病毒性疾病的暴发。     

母猪胎衣不下的病因与防治要点

猪的胎盘属于弥散型胎盘,这种胎盘的结构特点和饲养管理的不当,常常导致母猪胎衣不下发生,给生猪的生产繁殖带来极大损失。本文针对母猪胎衣不下发生病因、综合防治进行详细阐述,旨在对预防和治疗胎衣不下能有所帮助。