采用BSLB(Brine Shrimp Letality Bioassay)法对元宝枫 (Acer truncatum Bunge .)及同属植物活性部位的筛选(英文)

本文通过BSLB法对元宝枫及同属植物不同部位的水提物进行了活性试验 ,得知 :元宝枫的幼技、种皮的水提物具一定的生物致死活性 ;同属植物青榨槭叶和种子 ,秦岭槭、杈叶槭果翅的水提物也具一定的生物致死活性。

Toxicity of Fusarium solani Strains on Brine Shrimp (Artemia salina)

Discovery of anticancer drugs that must kill or disable tumor cells in the presence of normal cells without undue toxicity is an extraordinary challenge. Cytotoxicity of plant or fungal materials is considered as the presence of antitumor compounds. Brine shrimp lethality for larvae （nauplii） is used as prescreening test for the antitumor compounds. In this study, culture filtrates of eight strains of Fusarium solani isolated from seeds of various crops were tested for the toxic effect on brine shrimp. Five of the strains （TS, S-29, B-17, C-10, W-5） showed highest toxic effect and three of the strains （SR, T-9, L-25） showed low toxic activity on brine shrimp. Toxic activity reduced when culture filtrates were diluted. However, F. solani strains TS, B-17, SR, T-9 and L-25 caused more than 30% mortality at 1：10 dilution. Toxic activity was slightly reduced when the filtrates were neutralized with sodium hydroxide indicating possible role of pH of culture filtrate on toxicity. Lyophilized filtrates of these strains showed less activity as compared to un-lyophilized filtrates, n-Hexane soluble fraction was obtained only in three strains which showed mild toxicity whereas chloroform soluble fraction was obtained in negligible quantity and could not further be proceeded. Toxic effect of these strains showed variation from strain to strain. Compounds from F. solani could be exploited for the development of toxic compounds.

Anti-bacterial activity and brine shrimp lethality bioassay of methanolic extracts of fourteen different edible vegetables from Bangladesh

Objective:To investigate the antibacterial and cytotoxic activity of fourteen different edible vegetables methanolic extract from Bangladesh.Methods:The antibacterial activity was evaluated using disc diffusion assay method against 12 bacteria(both gram positive and gram negative).The plant extracts were also screened for cytotoxic activity using the brine shrimp lethality bioassay method and the lethal concentrations(LC_(50))were determined at confidence intervals by analyzing the data on a computer loaded with"Finney Programme??Results:All the vegetable extracts showed low to elevated levels of antibacterial activity against most of the tested strains(zone of inhibition=5-28 mm).The most active extract against all bacterial strains was from Xanthium indicum which showed remarkable antibacterial activity having the diameter of growth inhibition zone ranging from 12 to 28 mm followed by Alternanthera sessilis(zone of inhibition=6-21 mm).All extracts exhibited considerable general toxicity towards brine shrimps.The LC_(50)value of the tested extracts was within the range of 8.447 to 60.323μg/mL with respect to the positive control(vincristine sulphate)which was 0.91μg/mL.Among all studied extracts,Xanthium indicum displayed the highest cytotoxic effect with LC_(50)value of 8.447μg/mL.Conclusions:The results of the present investigation suggest that most of the studied plants are potentially good source of antibacterial and anticancer agents.

Diversity and connectedness of brine shrimp viruses in global hypersaline ecosystems

Brine shrimp(Artemia)has existed on Earth for 400 million years and has major ecological importance in hypersaline ecosystems.As a crucial live food in aquaculture,brine shrimp cysts have become one of the most important aquatic products traded worldwide.However,our understanding of the biodiversity,prevalence and global connectedness of viruses in brine shrimp is still very limited.A total of 143 batches of brine shrimp(belonging to seven species)cysts were collected from six continents including 21 countries and more than 100 geographic locations worldwide during 1977–2019.In total,55 novel RNA viruses were identified,which could be assigned to 18 different viral families and related clades.Eleven viruses were dsRNA viruses,16 were+ssRNA viruses,and 28 were−ssRNA viruses.Phylogenetic analyses of the RNA-directed RNA polymerase(RdRp)showed that brine shrimp viruses were often grouped with viruses isolated from other invertebrates and fungi.Remarkably,most brine shrimp viruses were related to those from different hosts that might feed on brine shrimp or share the same ecological niche.A notable case was the novel brine shrimp noda-like virus 3,which shared 79.25%(RdRp)and 63.88%(capsid proteins)amino acid identity with covert mortality nodavirus(CMNV)that may cause losses in aquaculture.In addition,both virome composition and phylogenetic analyses revealed global connectedness in certain brine shrimp viruses,particularly among Asia and Northern America.This highlights the incredible species diversity of viruses in these ancient species and provides essential data for the prevalence of RNA viruses in the global aquaculture industry.More broadly,these findings provide novel insights into the previously unrecognized RNA virosphere in hypersaline ecosystems worldwide and demonstrate that human activity might have driven the global connectedness of brine shrimp viruses.

BrineShrimp致死率生物鉴定法对八种民间药活性成分的研究

采用ＴｈｅＢｒｉｎｅＳｈｒｉｍｐＬｃｔａｌｉｔｙＢｉｏａｓｓａｙ法对民间药翼蓼ＰｌｅｒｏｘｙｇｏｎｕｍｇｉｒａｌｄｉｉＤｕｍｍｅｒｅｔＤｉｅｌｓ），金荞麦（Ｆａｇｏｐｙｒｕｍｄｉｂｏｔｒｙｓ（Ｄ．Ｄｏｎ）Ｈａｒａ），齿果酸模（ＲｕｍｅｘｄｅｎｔａｔｕｓＬ．）、巴天酸模（ＲｕｍｅｘｐａｔｉｅｎｔｉａＬ．）、网果酸模（ＲｕｍｅｘｄｉｃｔｙｏｃａｒｐｕｓＢｏｉｓｓｅｔＢｕｈｓｅ）、掌叶大黄（ＲｈｅｕｍｐａｌｍａｔｕｍＬｉｎｎ）、异叶败酱（ＰａｔｒｉｎｉａｈｅｔｅｒｏｐｈｙｌｌａＢｇｅ的甲醇提取物的石油醚、乙醚、乙酸乙酯、正丁醇等的萃取部分和水溶液部分及大自米（Ｎｏｔｈｏｌｉｒｉｏｎｈｙａｃｉｎｔｈｉｎｕｍ（Ｗｉｌｓ．）Ｓｉａｐｆ）生物碱总甙共３６种混合物进行了研究，测试出１４种活性成分，从而为抗癌药物的深入研究提供了科学依据。

Brine shrimp cytotoxic activities of Hippophae rhamnoides Linn leaves extracts

Objective:To evaluate brine shrimp lethality assay of solvent extracts(aqueous,methanol,ethanol,acetone,ethyl acetate,chloroform and n-hexane)of Hippophae rhamnoides(H.rhamnoides)leaves.Methods:Brine shrimp cytotoxicity assay was used to assess the cytotoxic potential of H.rhamnoides leaves extracts.Three vials for concentration of each extract were made and 10 shrimps per vial(30 shrimps per dilution)were transferred to specific concentration of each extract.Results:The mortality of aqueous extract was 46.7%,methanol extract was 46.7%,ethanolic extract was 50.0%,ethyl acetate was 26.7%,acetone extract was 33.3%,chloroform extract was 40.0%and n-hexane extract was 33.3%.The lowest LD50 was found in methanol extracts(1199.97μg/mL).Brine shrimp cytotoxicity of tested extracts of H.rhamnoides showed that mortality rate was concentration dependent.Conclusions:It is concluded that bioactive components are present in all leaves extracts of H.rhamnoides,which could be accounted for its pharmacological effects.Thus,the results support the uses of this plant species in traditional medicine.

Antimicrobial,antityrosinase and brine shrimp lethality test of Bauhinia rufescens Lam(Fabaceae)

Objective:To evaluate leaves and stem bark extracts of Bauhinia rufescens for antimicrobial activities,tyrosinase enzyme inhibition potentials and lethality to brine shrimp larvae(Artemiasalina).Methods:The plant materials were extracted based on polarity gradient of petroleum ether,ethyl acetate and methanol.The extracts were investigated for their antimicrobial,antityrosinase and cytotoxicity potentials.Results:The antimicrobial results indicated that the extracts inhibited the microbial growth in the range of 6.00-12.57 mm(450-1800μg/mL)for bacterial strains and 7.33-14.03 mm(225-1800μg/mL)for fungal strains.While the tyrosinase enzyme inhibition assay of the extracts showed that both the petroleum ether leaves and stem bark extracts exhibited a moderate inhibition(39.60%and 39.89%)towards the action of mushroom tyrosinase enzyme on L-DOPA.In the brine shrimp lethality assay,ethyl acetate and methanol extracts of the leaves of the plant were found toxic to the Artemia salina with IC_(50)values of 0.059 mg/mL and 0.389 mg/mL.However,both the petroleum ether and ethyl acetate extracts of the stem bark were nontoxic to the larvae.Conclusions:The result of the present study suggest the presence of phytochemicals possessing antimicrobial and antityrosinase activity as well as cytotoxic agents.

Studies on antimicrobial activity and brine shrimp lethality of crude samples of six different species of puffer ifshes

Objective:To evaluate the antimicrobial activity and brine shrimp lethality activity of six different species of puffer fishes,includingCyclichthys orbicularis,Diodon holocanthus,Canthigaster solandri,Arthron hispidus,A.inermis andLagocephalua inermis(L.inermis).Methodology:The puffer fishes were collected from Annangkovil Fish Landing Centre(Lattitude 11°30.47'N;Longitude 79°47.02'E),Parangipettai,Southeast Coast of India during summer season because of availability.Fresh tissue samples were collected from the clearly washed specimens,extracted with methanol at 37°C for 3 days and filtered through Whatman No.1 filter paper.The solvents such as methanol and ethanol were concentrated by using rotary evaporator under reduced pressure.The dark brown gummy mass was stored at 4°C for further analysis.Prepared crude samples were analysed with human pathogens to assess the antibacterial activity and this was carried out by using standard disc diffusion method.The brine shrimp lethality was calculated as the percentage of mortality which was firstly calculated by dividing the number of dead larvae by the total number and then multiplied to 100%.Results:The antibacterial activity of crude extract of puffer fishes were exhibited against 10 different human bacterial pathogens.Among the ten human pathogens,Arthron hispidus showed maximum zone of inhibition(8 mm)againstStaphylococcus aureus whileL.inermis showed minimum activity(1 mm)againstProteus mirabilis and no zone of inhibition was observed againstStaphylococcus aureus.Brine shrimp lethality was examined with six puffer fish extracts.Cyclichthys orbicularis showed maximum mortalities as 100%andL.inermis showed minimum mortalities as 70%at a concentration of 500μg/mL.Conclusion:In conclusion,the study showed the preliminary investigation of crude extracts of puffer fishes about the prominent activity against human bacterial pathogens.The extracts had a good cytotoxic potential against brine shrimpArtemina salina.

Phytochemical and toxicity evaluation of Phaleria macrocarpa(Scheff.)Boerl by MCF-7 cell line and brine shrimp lethality bioassay

Objective:To evaluate the cytotoxicity of Phaleria macrocarpa fruits extracts.Methods:The cytotoxicity test was carried out by in vitro MCF-7 cell line and in vivo brine shrimp lethality bioassay.Results:The preliminary phytochemical test showed the presence of alkaloids,carbohydrate,glycosides,saponin,terpene,steroids,phenols and flavonoids.The MTT-assay results showed that the highest percentage of cell viability was 106.23%at concentration of 1.25μL and the lowest percentage was 13.04%at concentration of 10μL.Conclusions:The MTT-assay and brine shrimp lethality bioassay results showed that the extract was non-toxic and it would be consumable as a herbal remedy.

THE DISCOVERY AND ITS GEOLOGICAL SIGNIFICANCE OF FOSSIL FAECAL PELLETS OF BRINE SHRIMPS IN MIRABILITE DEPOSIT

The brine shrimp, Artemia salina, is affiliated with a lineage of Arthropoda-class Crustacea-subclass Branchiopodaorder Anostraca-family Arlemiidae. The individuals, 10 to 11 mm long for male but slightly longer for female, show a striking adaptation to high-salinity waters and salt pans, for example, in Great Salt Lake of the U. S. A., a num-

废弃卤虫冻干工艺研究

本文以废弃卤虫为试验材料,采用电阻法测定废弃卤虫冻干过程的共晶点和共熔点温度,研究物料厚度、预冻温度、真空压力、冻干时间对废弃卤虫冻干品DHA含量和复水比的影响,并进行各工艺优化集成验证试验。结果表明,废弃卤虫冻干过程的共晶点和共熔点温度分别为-38.0、-26.9~-26.0℃,废弃卤虫最佳冻干工艺为物料厚度6 mm、-40℃预冻、真空压力70 Pa、冻干时间10 h,且加热板温度走势设置为0~40 min,由30℃上升至60℃,60~240 min稳定在60℃。在上述最佳工艺条件下,测得废弃卤虫冻干品DHA含量和复水比分别为3.1 g/kg、12.6。

咪唑类离子液体系列对卤虫的急性毒性研究

选取卤虫作为实验生物,研究了1-丁基,1-辛基,1-十二烷基-3甲基咪唑盐酸盐([Cnmim][Cl](n=4,8,12))3种离子液体的暴露对卤虫个体存活率的影响.同时,选取了实验室常用的有机试剂甲醇和乙腈,无机化学品重铬酸钾作为阳性对照.根据剂量效应曲线的拟合结果发现,受试离子液体系列随着咪唑环上烃基侧链碳原子数的增加,对卤虫的毒性效应也相应增强,3种离子液体对卤虫的LC50-24h依次为171.1,133.6,17.76μg/mL.离子液体对卤虫的LC50-24h比甲醇(LC50-24h=84.83mg/mL)和乙腈(LC50-24h=52.84mg/mL)低了2～3个数量级,重铬酸钾对卤虫的(LC50-24h=16.87μg/mL)与离子液体对卤虫的LC50-24h处于同一水平.

海洋微生物抗肿瘤活性菌株的分级组合筛选

为探索如何快速获取活性菌株的筛选模式 ,组合使用海虾生物致死法和tsFT2 10细胞的流式细胞术筛选模型 ,对从青岛前海和胶州湾潮间带海泥、动植物样品中分离的 162株海洋微生物 ,进行了抗肿瘤活性的分级组合筛选。经海虾生物致死法一级初筛 ,得到活性菌株 5 8株 (LC50 <5 0 0 μg/mL) ,阳性率为 3 5 .8%。对初筛有效的 5 8株菌利用流式细胞术筛选模型进行二级复筛 ,得到对tsFT2 10细胞有各种活性的活性菌株共 2 5株 ,阳性率为复筛菌株数的 43 .1%、初筛菌株总数的 15 .4% ,其中 ,诱发细胞坏死伴有凋亡诱导活性的菌株 7株 (占总菌株的 4.3 % ) ,具有细胞周期抑制活性的菌株 18株 (占总菌株的 11.1% )。该分级组合筛选模式与流式细胞术筛选模型的单独筛选模式相比 ,无漏筛 ,具有成本低、速度快、宜于大规模筛选等特点。

红树林来源内生真菌杂色曲霉Aspergillus versicolor MA-229次级代谢产物研究

从分离自红树林植物榄李新鲜叶片中的内生真菌杂色曲霉Aspergillus versicolor MA-229的发酵产物中分离得到新化合物1(22-epi-aflaquinolone B)和4个4-苯基-3,4-二氢喹啉酮类化合物(2~5)以及4个喹诺唑啉酮类化合物(6~9)。通过核磁共振和质谱等技术对化合物的结构进行了鉴定,并且通过X-射线单晶衍射实验确定了化合物1的绝对构型。这些化合物均表现出不同程度的卤虫致死活性和抗菌活性,其中新化合物1有明显的卤虫致死活性(LD50值为1.73μmol/L),与阳性对照活性相当。化合物1对农业病害菌小麦全蚀病菌(Gaeumannomyces graminis)也具有一定的抑制活性(MIC值为32μg/mL),化合物6对农业病害菌小麦赤霉病菌(Fusarium graminearum)有较好的抑制活性(MIC值为16μg/mL)。

海洋真菌杀虫活性的初步研究

对采集自浙江舟山海域底泥、福建省九龙江口红树林及福建省云霄县红树林共188株海洋真菌代谢产物的杀小杆线虫(Rhabditissp.)和卤虫(brine shrimp)活性进行初步研究.筛选出#164菌株代谢产物对小杆线虫显示了较好的杀害活性,其LC90小于0.5 mg/mL.#122菌株代谢产物对卤虫显示了较强的麻痹效应.#305菌株代谢产物对卤虫的LC50仅为75μg/mL.此外,对188株海洋真菌代谢产物的乙酰胆碱酯酶抑制活性进行研究,筛选出#229菌株代谢产物和#170菌株代谢产物具有较强的乙酰胆碱酯酶抑制活性,IC50分别为30μg/mL和25μg/mL,这些活性菌株具有产生杀线虫剂或乙酰胆碱酯酶抑制剂的特性.

中国卤虫卵孵化特性的研究

本文对山东埕口盐场、海南岛莺歌海盐场及青海柯柯盐湖天然采集的卤虫卵进行了孵化特性研究。埕口、莺歌海及柯柯的虫卵壳厚分别为9.7,11.4及13.2μm。在放入海水中进行孵化前上述三品系的卤虫卵分别在自来水中浸泡2,3及4小时能获得最佳孵化率。它们的最适孵化温度分别为25.6～28.5℃,27.0～30.0℃及27.0～33.0℃。最适孵化盐度分别为30,20及35％。孵化的同步性以“莺歌海”为最好,为6.4小时,“埕口”次之,为6.5小时,“柯柯”为7.4小时。 实验室内培养产卵后24小时内收集的虫卵用下列方法处理能终止虫卵的滞育状态:(1)埕口卤虫卵:浸泡于粗盐饱和溶液中2～3个月,孵化率可达91～94％。(2)莺歌海卤虫卵:浸泡于粗盐饱和溶液中2～5个月,孵化率可达80～91％。(3)柯柯卤虫卵:置于5℃2～3个月孵化率可达96～98％。用3％H_2O_2及3％甲醛溶液处理对终止滞育状态没有作用。天然采集的三个品系的卤虫卵用3％H_2O_2或3％甲醛溶液或用去壳溶液处理均可以提高孵化率。

利用盐田卤水池塘养殖卤虫的研究

1988年和1989年,作者在山东省盐场利用盐田卤水池进行了卤虫池塘养殖的研究。研究内容包括微藻培养试验、卤虫群体生长能力试验和间捕法养殖卤虫试验。通过试验在6个投饵池塘获得的鲜活卤虫产量为423.5—551.0公斤/亩(平均491.6公斤/亩),在4个不投饵池塘获得的产量为240.5—299.5公斤/亩(平均272.3公斤/亩)。本文报告了以上三个试验的内容和结果,并对卤虫养殖中出现的几个问题进行了讨论。

温度对青海尕海盐湖卤虫发育和种群增长的影响

1990～ 1999年对尕海卤虫的温度特性研究表明 ,无节幼虫的耐寒力比幼虫期高 - 1 7℃ ,比成虫期高- 0 9℃ ,即无节幼虫的耐寒力最强 ,幼虫对低温最敏感 ;孵化发育起始温度 9 94℃ ,幼虫发育起始温度10 33℃ ,有效积温 (℃·d)前者为 2 2 91± 2 0 8,后者为 2 6 1 2 6± 2 4 1,平均世代发育起点温度 10 2 8℃ ,有效积温 45 8 6 8± 5 7 6 0 ;依据生命表计算得出的各项生态学参数与温度存在显著的函数关系 ,其适温范围基本介于 10℃～ 39℃ ,最适温度介于 2 4 9℃～ 30 5℃ ,种群增长倍数 >2的温度范围在 15℃～ 34℃ ;依据 1993～ 1994和 1997年尕海水域温度测定资料 ,结合上述实验结果 ,得出该卤虫的年世代数为 2 6 7± 0 34个代 ,一年中有4 6 9± 0 43个生殖高峰期 ;最后一个完整世代的起始时间是 8月 10日 ;因积温不足 ,在 9月 1日以后出现的无节幼虫将不能完成到成虫期的发育 ;在 7月中旬至 9月中旬是种群生产力高峰期 ,在此期间种群的加倍时间 <30d ,内禀增长率 >0 0 2d-1,开发潜力最大。

6种地衣成分的海虾幼虫致死性生物测定

本研究采用海虾幼虫（ｂｒｉｎｅｓｈｒｉｍｐ）致死性生物测定法，对６种地衣的三氯甲烷提取物、单体化合物、多糖等１９个样品进行了抗癌活性的初步筛选。结果长松萝、太白花的三氯甲烷提取物，Ｒ－（＋）松萝酸、Ｓ－（－）松萝酸、红石耳Ⅱ具有较强的生物活性，Ｓ－（－）松萝酸钠活性最强ＬＣ＿（５０）为５．５μｇ／ｍｌ。６种地衣多糖均未显示生物活性。

南海深海链霉菌Streptomyces albiflaviniger SCSIO ZJ28中Elaiophylin的分离鉴定

从我国南海深海沉积物中分离得到一株放线菌SCSIO ZJ28,通过16S rRNA基因序列分析鉴定为Strep-tomyces albiflaviniger SCSIO ZJ28。对该菌株采用M-AM2ab培养基进行发酵,以溶剂萃取获得发酵提取物,通过卤虫致死活性、抑菌活性和HPLC-UV追踪,利用正相、反相硅胶柱层析和半制备高效液相色谱法分离,得到化合物1。经ESI-MS,1H及13C NMR,2D NMR和X-RD单晶衍射分析鉴定为大环内酯类化合物elaiophylin(1)。