Phlorizin alleviates deltamethrin-induced oxidative stress in brine shrimp Artemia

Deltamethrin(DEL),a commonly used pyrethroid pesticide,results in higher reactive oxygen species(ROS)levels in aquatic animals,which consequently unbalance the redox state.Phlorizin(PHL)is a flavonoid and a natural product promising to prevent or reduce pesticide-induced oxidative stress.Artemia is a micro-crustacean widely used in marine hatcheries and an experimental aquatic organism for environmental toxicology research.This research aimed to evaluate the toxicity of DEL on Artemia and the antioxidative effect of PHL against the toxicity.Results show that 0.08-mg/mL PHL exerted its antioxidative effects on hatching percentage of the cysts in 24-h incubation and on body length and survival rate of Artemia in 12-d culture.After 12-d culture,12-,24-,and 36-h DEL exposure showed significant drops in SOD,CAT,and GSH-Px enzyme activities,and significant increases in ROS and malondialdehyde(MDA)levels in Artemia(P<0.05).On the contrary,0.08-mg/mL PHL application improved the enzyme activities and decreased the ROS and MDA levels(P<0.05).Moreover,0.08-mg/mL PHL significantly increased mRNA expression levels of Cu/Zn SOD,CAT,GST,HO-1,NQO1,and Nrf2,and decreased mRNA expression level of Keap1 in the DEL-exposed Artemia(P<0.05).Therefore,DEL is toxic to Artemia,while PHL alleviates DEL-induced oxidative damage by possibly regulating the Nrf2signaling pathway.This study provided a theoretical basis for PHL to reduce pesticide-induced toxicity in aquatic animals.

Viability and hatchability of brine shrimp Artemia franciscana cysts after passing through the digestive system of eared grebes Podiceps nigricollis

Brine shrimp Artemia franciscana provide food for many migrating and staging birds that spend summer and fall on Great Salt Lake,Utah,USA.Artemia produce live young and cysts(hard-walled eggs);these cysts are commercially harvested on Great Salt Lake and support a large industry in Utah.It is unclear the impact that millions of hungry birds have on the Artemia population in the lake.To help assess that,this study evaluated cyst viability(percentage of cysts that contain an embryo)and hatchability(percent of cysts that hatch)from cysts that had passed through the digestive tract of eared grebes Podiceps nigricollis and cysts obtained directly from Great Salt Lake at the same site where each grebe was collected.Hatchability was significantly higher for cysts collected from the water column(19%)than from the stomach(0.3%)or intestines(3%)of eared grebes.Viability also was significantly different for cysts collected from the water column(29%),stomach(0.7%),and intestines(5%).These results indicate that eared grebes nutritionally benefit from eating cysts and that they may be an important food source for grebes in late fall after the adult population of Artemia dies off due to the water becoming too cold.Also,enough cysts survive their passage through the digestive system that grebes can vector hatchable cysts to other waterbodies.

Halomonas-PHB protects gnotobiotic Artemia against Vibrio and m odifies Artemia gut microbiota in xenic culture conditions

The prokaryotic cell storage compound ploy-β-hydroxybutyrate(PHB)has been considered as prebiotics that can be applied in aquaculture.In this paper,the dietary effect of a PHB-accumulating Halomonas strain(HM·PHB)identified from our previous work were studied in Artemia under gnotobiotic and xenic culture conditions,in comparison of Halomonas without PHB accumulation(HM)and microalgae Isochrysis(ISO)feeding.Under gnotobiotic condition,both HM·PHB and HM served as sole food supporting Artemia survival.Although both HM·PHB and HM feeding had no significant difference on Artemia survival percentage(P>0.05),HM·PHB significantly improved their resistance against Vibrio anguillarum challenge(P<0.05).Mass Artemia culture were further performed in xenic condition.Compared to ISO,HM·PHB feeding protected Artemia against V.anguillarum challenge(P<0.05),and HM·PHB and HM feeding resulted in increased T-AOC,pepsin,T-SOD and CAT activities(P<0.05).High throughput sequencing analysis showed that HM·PHB and HM feeding resulted in a lower Artemia gut microbial diversity(P<0.05),and modified the gut microbial community by remarkably reducing the Vibrio proportion.The outcome of the paper confirmed the beneficial effect of Halomonas-PHB in Artemia culture,which supports the use of Halomonas-PHB in the production of bio-secured live feed Artemia.

无效基因fruitless参与卤虫Artemia franciscana的生殖调控研究

fruitless(fru)是在昆虫与甲壳动物求偶、交配行为以及生殖发育过程中发挥关键作用的基因。卤虫不仅是水产育苗的重要开口饲料,而且是生物学研究的重要实验生物。本实验通过对卤虫Artemia franciscana转录组的筛选,获得了fru基因的开放阅读框(ORF)并进行生物信息学分析,利用qPCR技术研究了它在卤虫生殖腺发育的不同阶段的表达情况,最后利用RNAi显微注射技术,深入探索了它的功能。经过生物信息学分析,fru基因的ORF长度为1215 bp,其中包含了404个氨基酸,其分子量和等电点分别为45.19 kDa和5.28,属于酸性亲水性蛋白,且不含信号肽和跨膜结构;结构域预测显示fru存在BTB_POZ和HTH两个结构域,二级结构主要由α-螺旋和无规则卷组成,三级结构与其对应。qPCR结果显示,fru基因的表达量在卵囊的晚期胚胎中表现出明显的增加趋势,其表达量显著高于其他发育阶段(P<0.01);而在精巢的未成熟期,fru基因的表达量也有所增加,其表达量显著高于成熟早期、中期和晚期(P<0.01)。经过RNA干扰,我们发现fru基因的表达量有了显著的下降(P<0.01),并且经过受到干扰的雌虫后代都为休眠卵。这表明fru基因对于A.franciscana的生殖和发育有着重要的影响,甚至可能是影响卤虫繁殖方式的关键因素。通过本次研究,我们获得了关于fru基因在卤虫生殖发育中的作用及其相关分子机制的重要信息,可以帮助我们更好地理解这一重要的生物学过程。

光照强度与pH对卤虫卵(Artemia cysts)孵化率影响的响应面法分析

为进一步优化卤虫孵化条件、提高卤虫卵的潜在经济价值,试验采用中心组合旋转设计与响应面法分析光照强度与pH对卤虫卵孵化率的联合效应,利用Design-Expert 12.0分析得到最优试验组合并对预测结果进行验证。结果表明:光照强度与pH的一次效应对孵化率影响达到极显著水平(P<0.01),两者交互效应对孵化率影响显著(P<0.05),二次效应对孵化率影响极显著(P<0.01);随着pH与光照强度的升高,孵化率先升高后降低,越接近中心点孵化率变化量越小;光照强度为2 179 lx、pH为7.9为最适组合,最优孵化率为67.28%。对该试验组合进行验证,预测值与验证结果相符。综上,试验得到最适组合光照强度(2 179 lx)与p H(7.9),经试验验证在该条件下卤虫卵孵化率表现最优,经济效益最高。

pH对卤虫(Artemia saline)幼体急性毒性初步研究

采用半静水毒性试验法研究pH对卤虫(Artemia saline)幼体急性毒性及半致死毒性时间。通过概率单位法分析pH对卤虫的半致浓度及半致死毒性时间,并估算回归方程与95%置信区间。结果表明:pH对卤虫死亡数量有极显著影响(P<0.01),24、48 h的半致死pH及95%置信区间分别为:5.031(4.791~5.261)、5.345(5.227~5.467),安全pH为6.50;对应H+摩尔浓度分别为:9.311×10^(-6)、4.519×10^(-6)、3.192×10^(-7)mol/L。pH4.8、5.2、5.6、6.0的半致死时间及置信区间分别为22.275(14.286~28.928)、44.986(33.079~55.288)、89.813(79.244~100.231)、131.971(119.987~145.860)h。该研究为水质酸化对卤虫养殖、卤虫的低pH的耐受性研究及海水养殖中的水质管理提供相关参考依据。

山东盐田及新疆盐湖卤虫(Artemia parthenogenetica)生物学测定值的比较

从山东的盐场和新疆的盐湖采集了卤虫卵,进行了干燥、吸水和去壳的卵径、壳厚、无节幼体体长、染色体数目的生物学测定。按照文献[12]的分类,上述研究的卤虫,均属于第二类的孤雌生殖品系。盐湖的卤虫卵吸水和去壳后比盐场的卤虫卵大。而达板城东盐湖的无节幼体是最短的。各盐场的卤虫有2倍体、4倍体及5倍体,艾比湖的卤虫有2倍体和4倍体,达板城东盐湖只有4倍体。

温度对中国卤虫（Artemia sinica）种群的影响

本文采用生命表和数值模型分析等方法，对中国卤虫（Ａｒｔｅｍｉａｓｉｎｉｃａ）的温度特性做了研究。１．孵化发育起始温度９．０７℃，幼虫发育起始温度９．０７℃。有效积温（℃·ｄ）前者为０．９２１±０．９７９，后者为２０５．２６±２４．１，平均世代有效积温４５８．４±３９．６。２．各项生命表参数均与温度呈显著的函数关系，其适温范围基本介于１８─３４℃，最适点介于２６．５─２８．５℃，种群增长倍数＞２的温度范围在１８─３４℃。３．在山西盐池水域，该卤虫的年世代数为６．２±０．６２个世代、第２─５个世代是种群生产力高峰期。在此期间种群的加倍时间＜１５ｄ．内禀增长率＞０．０４ｄ￣（－１）。

THE BRINE SHRIMP (ARTEMIA PARTHENOGENETICA) AS ENCAPSULATION ORGANISM FOR PROPHYLACTIC CHEMOTHERAPY OF FISH AND PRAWN

Brine shrimp (Artrmia parthenogenetica) which had ingested three water-insoluble antibactrial drugs ie.sulfadiazine(SD), oxytetracycline (OTO) and erythromycin estolate (ERY-Es) were fed to TiIapia andMysis Ⅲ of Penaeus orientalis K..The drug contentS in the predators were then determined. Aferadministration of drugs to TiIapia and Mysis Ⅲ, through the bio-encapsulation of the brine shrimp,efficacious therapeuical concenration of OTC and ERY-Es(but not SD) in the pndators could bereached and meintained for mere than 8 hours.

Effect of carbon and nitrogen ratio control on Artemia growth, water quality, biofloc microbial diversity under high salinity and zero-water exchange culture condition

Biofloc technology has been applied successfully in the intensive aquaculture of several fish and shrimp species. The growth of heterotrophic microorganisms can be stimulated through adding extra carbon, which reduces the nitrogen level in the water and provides microbial protein to the animals. However, most of the studies and practical applications have been conducted in freshwater and marine environment. This paper focused on brine shrimp Artemia that lives in high salinity environment together with other halophilic or halotolerant microorganisms. The effect of carbon supplementation on Artemia growth, water quality, and microbial diversity of biofl ocs was studied in the closed culture condition without any water exchange. The salinity of the culture medium was 100. A 24-d culture trial was conducted through supplementing sucrose at carbon/nitrogen (C/N) ratio of 5, 15, and 30 (Su5, Su15, and Su30), respectively. The culture without adding sucrose was used as a control. Artemia was fed formulated feed at a feeding ration of 60% recommended feeding level. The results showed that sucrose supplementation at higher C/N ratio (15 and 30) signifi cantly improved the Artemia survival, growth and water quality ( P <0.05). Addition of sucrose at C/N ratio of 15 and 30 significantly increased biofloc volume (BFV)( P <0.05). The Illumina MiSeq sequencing analysis showed that supplementing carbon at C/N ratio of 15 had a better total bacterial diversity and richness, and shaped the microbial composition at genera level. This study should provide information for studying the mechanism of biofloc technology and its application in high salinity culture conditions.

Piwi基因参与两性卤虫(Artemia franciscana)的生殖调控研究

Piwi(P-element induced wimpy testis)基因编码Piwi蛋白,在生殖干细胞的自我更新、减数分裂过程、RNA沉默和转录调控中起重要作用。为探究Piwi基因参与两性卤虫生殖发育的作用,从两性卤虫Artemia franciscana转录组中筛选获得Piwi基因开放阅读框,进行序列分析和结构域预测等生物信息学分析,采用qPCR技术研究该基因在A.franciscana生殖腺发育不同时期表达特征,并利用RNAi显微注射技术探究其功能。生物信息学分析表明,A.franciscana Piwi基因的开放阅读框长2619 bp,编码872个氨基酸;Piwi基因编码的蛋白分子量为98.11 kDa,等电点为9.50,为碱性亲水性蛋白,无信号肽和跨膜结构;存在Piwi和PAZ结构域及ArgoL1结构域,二级结构以α-螺旋为主,三级结构与之对应;系统进化树显示A.franciscana与蚤状溞和大型溞的Piwi序列相似性最高。qPCR结果表明,Piwi基因在卵巢的卵黄发生晚期和晚期胚胎表达量最高,显著高于卵黄发生早期和早期胚胎(P<0.01);Piwi基因在精巢的未成熟期表达量最高,显著高于成熟早期、成熟中期和成熟晚期(P<0.01)。RNAi结果显示,该方法能显著降低Piwi基因的表达水平(P<0.01),并导致A.franciscana所产后代均为休眠卵,说明Piwi基因不但在A.franciscana生殖发育调控起重要作用,而且可能在其繁殖方式决定过程中起关键作用。研究结果为两性卤虫Piwi/piRNA功能和分子机制调控的研究提供了基础信息,将有助于揭示Piwi基因参与调控两性卤虫生殖发育机制。

EFFECT OF ARTIFICIAL REGULATIONS OF ARTEMIA n-3 HUFA CONTENT ON GROWTH AND SURVIVAL OF BLACK SEABREAM (SPARUS MACROCEPHLUS) LARVAE

The requirement for dietary n-3 HUFA (highly unsaturated fatty acid ) for growth and survival of black seabream (Sparus macrocephalus) larvae was tested using Artemia with various levels of n-3 HUFA. Four tueatments with Artemia differing in theirn-3 HUFA were prepared by feeding them various oil emulsions .The results indicated that dietary n-3 HUFA significantly influences fish n-3 HUFA levels and are necessary for good growth and survival of black seabream larvae . The proper value of n-3 HUFA level and DHA, EPA level in Artemia should be 4.273% and 0.873% (wet weight) for good growth and survival in black seabream larvae respectively.

The critical stage for inducing oviparity and embryonic diapause in parthenogenetic Artemia(Crustacea: Anostraca): an experimental study

The brine shrimp Artemia exhibits two reproductive modes: 1) oviparity, producing diapause embryos;and 2) ovoviviparity, producing free-swimming nauplii. Previous studies have suggested the existence of a critical stage that determines the reproductive mode. Physicochemical factors, such as photoperiod, temperature, and salinity, have been suggested to irreversibly aff ect the reproductive mode of oocytes during this critical stage. In this study, experiments were carried out using a photoperiod and temperature-sensitive parthenogenetic Artemia clone where maternal Artemia were shifted bidirectionally between ovoviviparity (18 h L:6 h D, 27℃) and oviparity (6 h L:18 h D, 19℃) culture conditions. In the main experiment ( Artemia shifted at six diff erent stages including the post-larva Ⅱ to adult Ⅱ), the reproductive mode of fi rst brood was converted when shifting was performed on post-larva Ⅱ and Ⅲ but was not converted when females were shifted after post-larva Ⅲ. A supplementary experiment further revealed that the reproductive mode of fi rst brood could be altered when shifting females at an "early phase of postlarva Ⅳ", characterized by a developing ovisac reaching the middle of the third abdominal segment, ventral spines, and some oocytes growing larger than the others. In both experiments, reproductive modes of the second brood were signifi cantly aff ected when the shifting was performed on post-larva Ⅳ. These results suggest that the critical stage for inducing oviparity and embryonic diapause is at the previtellogenic stage of oocytes, or at maternal "early phase of post-larva Ⅳ" for the first-brood off spring. During this stage, diff erential gene expression patterns of the two destined oocytes may be triggered by the token stimuli signals received by the oocytes.

Eff ect of polystyrene microplastics and temperature on growth,intestinal histology and immune responses of brine shrimp Artemia franciscana

Microplastics pollution and seawater temperature rise have been the major environmental issues,threatening the survival and biodiversity of marine organisms.This study evaluated the combined effect of temperature and polystyrene microplastics(MP)on Artemia,a filter-feeding crustacean that is widely used for environmental toxicology studies.Brine shrimp Artemia franciscana were exposed to three MP concentrations(0,0.2,and 2.0 mg/L)and three temperatures(22,26,and 30℃)for 14 d.In general,higher MP concentration and temperature led to a decreased survival rate and growth.Two-way ANOVA analysis indicated that the survival rate of Artemia was significantly impacted by both MP concentration and temperature(P<0.05),but there was no significant interaction between two factors(P>0.05).Growth of Artemia was significantly impacted by temperature(P<0.05),and with a significant interaction between two factors(P<0.05).Furthermore,the enzymatic activity,intestinal histological analyses,and immune gene expression were determined for Artemia reared at 30℃ with three MP concentrations(0,0.2,and 2.0 mg/L).The results showed that 2.0-mg/L MP resulted in reduced Artemia intestinal microvilli and exfoliated epithelia cells,significantly increased acid phosphatase(ACP)activity(P<0.05)and immunerelated gene ADRA1 B and CREB3 expression,revealing that higher MP concentration could induce oxidative and immunological stress on Artemia at 30℃.Overall,our study suggests that MP and temperature have combined adverse effect on Artemia,especially at relatively high temperature and polystyrene MP concentration.These findings are important to understand the potential ecological risks posed by these two factors on the organisms in marine environment.

Results of an Intentional Introduction of Artemia sinica in the High-Altitude Tibetan Lake Dangxiong Co:On a Base of Surveys in 2011 and 2013

1 Introduction The human population is projected to reach 9200million by 2050.A fundamental question for science is whether it is possible to increase food production to meet the demands of a human population of that magnitude.It’s possible only to increase an aquaculture production.Cultivation of fish/shrimp larvae is a bottleneck in a cultivation of the different organisms.Live food

Microbiota of Salt Lakes as Food for the Brine Shrimp Artemia

The brine shrimp Artemia is often abundantly found in hypersaline environments.The larvae(‘nauplii’)hatching from its dormant embryos(‘cysts’)are a crucial live food item for the larval stages of fish and shrimp in aquaculture

The Biodiversity of Shrimp Genus Artemia from Russian Lakes:Morphometric,Cytogenetics and DNA-analysis

Shrimps of genus Artemia are the inhabitants of continental and marine waters with salinity of 70 to 350 g/l and above.Artemia is able to survive in the conditions in which other animals cannot exist.This is due to adaptations:effective osmoregulation system,the ability to synthesize of respiratory pigment(hemoglobin)and diapauses cysts(Litvinenko at.al.,2009).Cysts of this

中国卤虫(Artemia sinica)早期胚胎发育相关基因表达序列标签(ESTs)分析

以mRNA差异显示技术分析了中国两性生殖卤虫早期胚胎发育相关基因的表达序列标签(ESTs),共回收了2000条差异条带并进行了克隆、测序,在GenBank/DDBJ/EMBL数据库中用BLASTX和BLASTN软件分别与NCBI上的非冗余核酸序列数据库进行同源性比对,得到分值较高、长度在100bp—2kb以内的ESTs序列133条。所发现的133条ESTs的生物学功能分类为:能量代谢占24%、细胞生长、分裂相关基因占23%、RNA相关占17%、体节形成占12%、蛋白质合成占3%、转录因子占3%、离子转运占5%、细胞信号转导占4%、未知占9%。对所发现的ESTs进行了RT-PCR测定,排除假阳性能够提供足够数量的信息用于获取其全长cDNA,进而获得其全长基因序列以用于功能研究。另外,研究发现了包括小热休克蛋白基因家族、Hox基因家族的abdA和Dfd基因、A-trh基因(排盐器官及盐调节基因)、HIF(缺氧诱导因子)、CK-M2(肌肉型磷酸激酶基因)和GRP基因(富甘氨酸蛋白基因)等20个具有明显功能的相关基因的ESTs和完整基因序列和开放阅读框(ORF)。经初步推断这些ESTs和基因的功能大多为与卤虫发育的生理、生化相关的基因有关。所研究的133条ESTs基本代表了卤虫早期胚胎发育相关功能基因的表达谱。

肼与苯肼对卤虫Artemia Salina的毒性研究

采用卤虫对肼与苯肼的毒性作用进行了测定,结果表明苯肼的毒性明显大于肼.肼24hLC_(50)值为0.167mg/L(0.153～0.180mg/L),48hLC_(50)值为0.088mg/L(0.068～0.120mg/L),苯肼24hLC_(50)值和48hLC_(50)值分别为0.041mg/L(0.039～0.042mg/L),及0.015mg/L(0.0114～0.0202mg/L),结果表明卤虫生物测试技术在海水监测中是一个敏感、快速和廉价的方法.

Genetic Parameters of Body Length and Response to Selection for Growth Across Four Generations of Artemia sinica

To investigate the genetic components of growth in the brine shrimp Artemia sinica, we estimated the genetic parameters of body length and the response to selection using a fully pedigreed population of A. sinica. The base population was generated from four wild founder populations. We tested 4160 offspring in 360 families over four generations for growth and survival performance. Across four generations, we produced full-and half-sib families with nested mating, where two dams were mated to the same sire. Individual body length was measured for each nauplius at day 20 post-hatching. Heritability of body length was estimated across four generations with the restricted maximum likelihood method. The heritability of body length in A. sinica was low(0.14 ± 0.05), and the common environmental effect was 0.14 ± 0.02. We estimated the response to selection for body length by calculating the difference in the mean breeding values between different generations. The accumulated genetic gain in body length was 278.94 μm after three generations of selection. This low response to selection was probably caused by the low heritability of body length, small sample size, and the low selection intensity(50%). The results suggest that A. sinica selective breeding programs must be changed to generate any substantial, sustainable genetic increases in body length. We suggest that optimal genetic gains could be achieved by introducing wild strains into the nuclear breeding population to increase genetic variation, and by increasing the size of the breeding population to allow for increased selection intensity.