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近年来,环境DNA(Environmental DNA, eDNA)技术作为一种新的水生生物调查方法发展迅速,在水生生态系统的研究领域被广泛应用到物种检测、生物多样性评价、生物量评估等方面。然而,很少有研究专门评价eDNA技术操作流程中不同的eDNA富集方法与提取方法对研究结果的影响,从而针对具体研究对象建立一套最佳的eDNA技术操作流程。此外,由于物种间生活习性的差异,不同物种释放到环境中的DNA量及DNA片段大小不同,因而,针对不同研究对象需采用不同的eDNA富集与提取方法。本研究以中国对虾(Fenneropenaeus chinensis)为研究对象,采用滤膜法富集eDNA,结合血液与组织DNA提取试剂盒提取eDNA。选取直径为47 mm的玻璃纤维膜、硝酸纤维膜、聚碳酸酯膜、尼龙膜共4种材质的滤膜,每种滤膜根据其孔径大小设置0.45、0.8、1.2、5 μm共4个梯度,取样水量设置500 ml、1 L、2 L共3个梯度。结果显示,滤膜材质、滤膜孔径大小及取样水体体积均对中国对虾的定性与定量分析具有一定的影响,其中,0.45 μm的玻璃纤维滤膜过滤2 L水样能够检测到的DNA拷贝数最多,并依据此建立了一套中国对虾eDNA技术的操作流程,提高了中国对虾的检出率,为后续中国对虾的分布监测及生物量评估提供了基础。 相似文献
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近年来,环境DNA(Environmental DNA, eDNA)技术作为一种新的水生生物调查方法发展迅速,在水生生态系统的研究领域被广泛应用到物种检测、生物多样性评价、生物量评估等方面。然而,很少有研究专门评价e DNA技术操作流程中不同的eDNA富集方法与提取方法对研究结果的影响,从而针对具体研究对象建立一套最佳的eDNA技术操作流程。此外,由于物种间生活习性的差异,不同物种释放到环境中的DNA量及DNA片段大小不同,因而,针对不同研究对象需采用不同的eDNA富集与提取方法。本研究以中国对虾(Fenneropenaeus chinensis)为研究对象,采用滤膜法富集eDNA,结合血液与组织DNA提取试剂盒提取e DNA。选取直径为47 mm的玻璃纤维膜、硝酸纤维膜、聚碳酸酯膜、尼龙膜共4种材质的滤膜,每种滤膜根据其孔径大小设置0.45、0.8、1.2、5μm共4个梯度,取样水量设置500 ml、1 L、2 L共3个梯度。结果显示,滤膜材质、滤膜孔径大小及取样水体体积均对中国对虾的定性与定量分析具有一定的影响,其中,0.45μm的玻璃纤维滤膜过滤2 L水样能够检测到的DNA拷贝数最多,并依据此建立了一套中国对虾e DNA技术的操作流程,提高了中国对虾的检出率,为后续中国对虾的分布监测及生物量评估提供了基础。 相似文献
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随着人类活动的影响,鱼类资源正在急剧下降,开展鱼类监测对于鱼类多样性的保护具有非常重要的意义。传统的鱼类监测方法因具有破坏性、调查者也要专业形态学知识、调查工作量大等弊端,故亟需一种新的技术方法进行辅助补充。环境DNA （eDNA）技术的发展使得鱼类调查更加的省时省力、节约成本、无损伤,目前在鱼类的监测中广泛使用,然而eDNA技术也存在着一定的缺陷所以不能完全取代传统方法。eDNA技术包含了样品的采集及处理、eDNA提取、eDNA扩增、测序和生物信息学分析几个主要步骤,在整个流程中每一个步骤都非常必要,对单个步骤又有不同的实验方案,选择不同对鱼类eDNA的检测效率也会产生重要的影响。目前国外对于eDNA技术应用于鱼类监测所研究的问题较为丰富而全面,如生物多样性监测、入侵物种检测、濒危物种检测和生物量的估算等,虽然国内的起步不晚,但国内对此研究方向较为单一,需要进一步重视。 相似文献
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长江中下游干流环境DNA样本鱼类物种检测的初步研究 总被引：1，自引：0，他引：1
为开发适用于长江鱼类的环境DNA检测体系,在长江中下游干流24个采样点同步采集水样,抽滤后提取环境DNA,利用线粒体细胞色素B简并引物进行PCR扩增,扩增产物克隆测序得到419条序列,通过在Gen Bank核酸序列数据库进行BLAST序列比对确定物种信息,从来源于17个采样点的115条匹配成功的序列中,共检测到10种鱼类序列,代表15种鱼类。 相似文献
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大型河流中鱼类组成的eDNA监测效率:以长江武汉江段为例 总被引：2，自引：0，他引：2
为了探讨大型河流中的 eDNA 监测效率, 以长江为大型河流的代表, 以鱼类为水生生物的代表, 分析传统捕捞监测和 eDNA 监测结果的差异, 研究 eDNA 监测技术对长江武汉江段鱼类组成的监测能力、监测效率、平行样设置等问题。结果显示: (1) 用 1 对 eDNA 宏条形码引物(mlCOIintF/jgHCO2198R)共监测到 89 种鱼类, 其中 30 种可与历史捕捞调查记录互相确认, 另外 59 种需要更完善的条形码数据库来解决序列比对注释问题; (2) 9 月在武汉监测断面, 可用单引物(mlCOIintF/jgHCO2198R) eDNA 监测到的物种最优估计约 99 种, 单样品 eDNA 监测的鱼类物种检出能力约为 26 种, 检出效率约为 25.8%; (3) 在 80%的检出度目标下, 需要约 10 个平行样, 在 95%的检出度目标下, 需要约 17 个平行样。本研究在长江武汉江段鱼类 eDNA 监测效率和平行样设置的相关量化结果可为长江其他断面及其他大型河流的 eDNA 监测提供量化参考。同时, 本研究表明, 更完善的 DNA 宏条形码数据库和合适的平行样设置是未来 eDNA 监测作为常规监测手段进行运用的前提。 相似文献
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浮游植物作为水生生态系统的重要组成部分，其群落和分布特征可以反映水体的环境变化和营养状态。环境DNA宏条形码技术将传统的基于领域的生态学与深入的分子方法和先进的计算工具结合在一起，这在生物多样性的研究中具有巨大的优势。本研究旨在采用环境DNA宏条形码技术对鄱阳湖浮游植物多样性的组成及空间分布进行研究。于2019年的4月采集鄱阳湖环境水样，每个样点重复3次采集。使用0.45 μm的混合纤维素滤膜对水样进行真空抽滤。用试剂盒DNeasy? Blood & Tissue Kit进行滤膜DNA的提取，使用通用引物1380F：5’-TCCCTGCCHTTTGTACACAC-3’和1510R：5’-CCTTCYGCAGGTTCACCTAC-3’针对真核浮游植物 18S rDNA 基因的 V9 区域进行PCR扩增，高通量测序并结合生物信息学技术分析鄱阳湖浮游植物的群落组成。基于环境DNA宏条形码技术鉴定到浮游植物10门24纲54目101科166属，其中绿藻门和硅藻门种类较为丰富，检测到的属明显多于传统方法检测到的。中部区域的浮游植物群落多样性和均匀度较高，且鄱阳湖浮游植物整体丰富度较高。NMDS分析阐明，中部和北部区域( p = 0.004)、南部和北部区域之间群落结构差异显著( p = 0.011)。冗余分析表明，叶绿素a (Chl-a)、pH、流速(V)对各区域的浮游植物群落影响较显著。本研究使用环境DNA宏条形码技术评估了鄱阳湖浮游植物生物多样性，证明了环境DNA宏条形码技术检测浮游植物群落结构的可行性。环境DNA宏条形码作为一种新兴的生物多样性监测手段，其可用于快速检测鄱阳湖浮游植物生物多样性及其空间分布，为鄱阳湖生物多样性监测以及生态系统健康评估提供新的技术手段。 相似文献
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应用环境DNA技术对邵伯湖浮游动物物种检测的初步研究 总被引：1，自引：0，他引：1
为掌握邵伯湖浮游动物物种组成,利用环境DNA技术于2018年11月对其浮游动物群落进行了监测。在邵伯湖15个采样点同步进行了水样采集,抽滤后进行了DNA提取,利用线粒体DNA 12S r RNA引物进行PCR扩增,并进行了高通量测序。结果显示,从二代测序的1114482条DNA序列中,比对得到浮游动物OTUs数68个。利用宏条形码共检出浮游动物22种(隶属于14科18属),邵伯湖浮游动物功能群以轮虫滤食者RF为主,各样点浮游动物α和β多样性的各项指数较为均匀,表明邵伯湖各区域的浮游动物多样性水平较接近。环境DNA技术适用于浅水湖泊浮游动物群落监测,在现阶段的淡水浮游动物资源监测中,该技术宜与传统的监测方法结合使用。 相似文献
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准确掌握物种分布和种群动态是渔业资源评估的基础。然而,对某些种群规模小或生活史复杂的物种进行监测的难度很大。近年来,环境DNA技术快速兴起,已被广泛应用于各类物种监测、生物多样性评估和生物量评价等领域。为了解冬季中国对虾(Fenneropenaeus chinensis)越冬洄游过程中的分布情况,2019年12月在黄海中南部采集表、中、底3个水层水样,检测其中中国对虾的eDNA,并对表层沉积物进行室内实验。结果显示,中国对虾eDNA在自然水体中呈现特殊的垂直分布规律:底层浓度高,表层浓度低,这一规律与中国对虾生活习性相关;表层沉积物会在外力作用下再悬浮,并向周围释放eDNA,对水体造成较大程度影响,本研究将采集到的表层沉积物分为3个实验组,3个实验组最大释放量分别为1624.06、3453.34和1143.24 copies/L,沉积物对水体的影响持续1周左右。 相似文献
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This study was conducted to gain insight into the characteristics of algal blooms in relation to the marine environment of
Gamak Bay. From the first known occurrence of algal blooms in 1984 until 2006, 23 causal species have been identified, the
most common ones being Prorocentrum sp., Cochlodinium polykrikoides, Chaetoceros sp., Skeletonema costatum, and Heterosigma akashiwo. A principal component analysis indicated that blooms of these species develop under different conditions in terms of water
temperature, salinity, precipitation, and insolation. Field data showed Cochlodinium polykrikoides to be favored by high temperatures and to be euryhaline, whereas Skeletonema costatum appeared to be eurythermal and euryhaline. Prorocentrum sp. and Chaetoceros sp. appeared to be stenothermal and stenohaline. Finally, Heterosigma akashiwo appeared at the lowest temperature and highest salinity of the five species and was classified here as stenothermal and stenohaline. 相似文献
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根据2015年3-5月在青岛斋堂岛附近海域使用张网采集的1250尾薛氏海龙(Syngnathus schlegeli)样本,分析了该水域薛氏海龙的性比、副性征、性腺指数、性腺成熟度与卵径、育儿袋中胚胎等繁殖生物学特征。结果表明,青岛斋堂岛附近海域薛氏海龙春季繁殖种群雌雄比例分别为63.28%和36.24%,未成年个体占0.48%,雌雄比为1.75:1,明显偏离1:1的性别比例;繁殖群体中雄性个体的性腺指数始终保持在较低水平,为0.01%~4.85%,雌性个体相对较高,为0.05%~128.99%,二者差异极显著(P0.01);组织学观察发现,性腺在3—5月份迅速发育成熟,雄性排精后精巢腔中仍剩余较多精细胞,表明雄性个体为多次排精;雌性Ⅲ~Ⅳ期卵巢中均分布有不同时相的卵子,且卵径分布呈现两个峰值,表明雌性个体为分批产卵;根据雌性个体V期卵巢中成熟卵子的卵粒数与雄性个体育儿袋中的平均怀卵数基本相同,育儿袋中胚胎的发育阶段也基本一致,推测其交配模式可能为一雄一雌制,抑或是雄性在同一天内与多个雌性交配的一雄多雌制。斋堂岛附近海域薛氏海龙的交配模式还有待室内行为观察并结合微卫星标记等手段进一步确认。 相似文献
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T. Citarasu K. Venkatramalingam M. Micheal Babu R. Raja Jeya Sekar M. Petermarian 《Aquaculture International》2003,11(6):581-595
The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could
lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment
in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 103 to 3.76 × 105 cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load
in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 105 cfu/g).
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
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东太平洋公海茎柔鱼种群遗传结构初步研究 总被引：8，自引：1，他引：7
根据2009年7月到10月我国大型鱿钓渔船在东太平洋(81.9°～94.9°W,8.9°N～11.3°S)秘鲁和哥斯达黎加外海作业期间采集的样本,利用线粒体DNA细胞色素b基因的724 bp部分序列,分析了采自秘鲁7个采样点和哥斯达黎加3个采样点的155个个体的序列多样性与种群遗传结构.结果显示,724 bp片段中发现了16个变异位点,155个个体出现43个单倍型.序列多样性分析结果揭示,155个个体的平均单倍型多样性指数为0.873,核苷酸多样性指数为0.003 69.群体间共享6个单倍型,秘鲁外海茎柔鱼群体享有最多特有单倍型(20个).分子方差分析揭示,82.70％的遗传变异性出现在种群内;群体间的FST分析揭示部分秘鲁群体与哥斯达黎加群体,部分秘鲁群体间存在着显著的遗传分化;而哥斯达黎加群体内部不存在显著的遗传分化. 相似文献
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Garth L. Fletcher Madonna J. King Ming H. Kao Margaret A. Shears 《Fish physiology and biochemistry》1989,6(2):121-127
Several species of marine teleosts have evolved blood plasma antifreeze polypeptides which enable them to survive in ice-laden seawater. Four distinct antifreeze protein classes differing in carbohydrate content, amino acid composition, protein sequence and secondary structure are currently known. Although all of these antifreezes are relatively small (2.6–33 kd) it was generally thought that they were excluded from the urine by a variety of glomerular mechanisms. In the present study antifreeze polypeptides were found in the bladder urine of winter flounder (Pseudopleuronectes americanus), sea raven (Hemitripterus americanus), ocean pout (Macrozoarces americanus) and Atlantic cod (Gadus morhua). Since the plasma of each of these fish contains a different antifreeze class it would appear that all four classes of antifreeze can enter the urine. The major antifreeze components in the urine of winter flounder were found to be identical to the major plasma components in terms of high performance liquid chromatography retention times and amino acid composition. It is concluded that plasma antifreeze peptides need not be chemically modified before they can enter the urine. 相似文献
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Use of ongrownArtemia in nursery culturing of the tiger shrimp 总被引：1，自引：0，他引：1
Juvenile and adultArtemia produced in a semi flow-through culture system were used as food for postlarval shrimp. The growth performance of shrimp reared on such ongrownArtemia live prey is identical to the growth obtained when feeding newly hatchedArtemia. However, a significantly better stress resistance is obtained when the postlarvae are exposed to a low salinity in a stress test. Besides nutritional and energetic advantages, the use ofArtemia biomass for feeding postlarval shrimp also results in improved economics as expenses for cysts and weaning diets can be reduced. 相似文献
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为抑制萱藻丝状体保存和扩增过程中出现的小伪菱形藻与碎片菱形藻的生长,本实验应用实验生态学方法,分别建立了丝状体与小伪菱形藻、丝状体与碎片菱形藻、丝状体与小伪菱形藻和碎片菱形藻的共培养体系,研究了1.00～4.00μg/mL二氧化锗(GeO_2)对共培养条件下丝状体生长发育及附生硅藻生长的影响。结果显示:(1)处理萱藻丝状体和硅藻共培养体系的适宜GeO_2浓度为1.00～2.50μg/mL,各实验组14 d的硅藻抑制率均高于67.33%±5.18%,且丝状体生长发育良好,2.00μg/mL为最适浓度,此浓度下丝状体日均增长率最高,在各培养体系中均大于11.00%,且诱导后孢子囊枝比例和孢子囊直径分别为57.47%±5.31%和(24.55±1.01)μm,与对照组差异不显著;(2) 3.50和4.00μg/mL GeO_2虽对硅藻抑制效果更佳,但同时也会抑制丝状体生长和后期孢子囊的形成与发育,其中4.00μg/mL GeO_2可导致丝状体死亡;(3)碎片菱形藻较小伪菱形藻对GeO_2更敏感。实验14 d,各浓度GeO_2对碎片菱形藻的抑制率为(82.10%±2.40%)～(96.35%±0.79%),均高于同浓度GeO_2对小伪菱形藻的抑制率;同时在丝状体与小伪菱形藻和碎片菱形藻的共培养体系中,碎片菱形藻占硅藻比例随GeO_2浓度升高而相应下降。 相似文献
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Araceli Hipólito-Morales A. M. Maeda-Martínez S. F. Martínez-Díaz 《Aquaculture International》2009,17(1):85-90
The effect of Microbacterium sp. strain 8L and Exiguobacterium mexicanum strain 8N was evaluated in the diet of Artemia under xenic conditions. Viable cultures of bacteria were provided to xenic cultures of Artemia in combination with Sacharomyces cerevisiae, cornflour or Spirulina, and the effect on the survival and growth was recorded. The use of these bacterial strains improves significantly the survival
of Artemia independently of the used food (P < 0.05), and variable results were observed in the growth. 相似文献
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G. Immanuel T. Citarasu V. Sivaram M. Michael Babu A. Palavesam 《Aquaculture International》2007,15(2):137-152
One of the major problems in the shrimp culture industry is the difficulty in producing high-quality shrimp larvae. In larviculture,
quality feeds containing a high content of highly unsaturated fatty acids (HUFA) and ingredients that stimulate stress and
disease resistance are essential to produce healthy shrimp larvae. In the present study, Penaeus monodon postlarvae (PL15) were fed for 25 days on an unenriched Artemia diet (control; A) or on a diet of Artemia enriched with either HUFA-rich liver oil of the trash fish Odonus niger (B), probionts [Lactobacillus acidophilus (C1) or yeast-Saccharomyces cerevisiae (C2)] or biomedicinal herbal products (D) that have anti-stress, growth-promoting and anti-microbial characteristics. P. monodon postlarvae fed unenriched Artemia exhibited the lowest weight gain (227.9 ± 8.30 mg) and specific growth rate (9.95 ± 0.05%), while those fed the HUFA-enriched
Artemia (B) exhibited the highest weight gain and specific growth rate (362.34 ± 12.56 mg and 11.77 ± 0.08%, respectively). At the
end of the 25-day rearing experiment, the shrimp postlarvae (PL40) were subjected to a salinity stress study. At both low
and high (0 and 50‰) salinities, the group fed the control diet (A) experienced the highest cumulative mortality indices (CMI)
935.7 ± 2.1 and 1270.7 ± 3.1, respectively. Those fed diet D showed the lowest stress-induced mortality, and CMI were reduced
by 31.1 and 32.3% under conditions of low and high salinity stress, respectively. A 10-day disease challenge test was conducted
with the P. monodon postlarvae (PL40–PL50) by inoculating the shrimp with the pathogen Vibrio harveyi at the rate of 105–107 CFU/ml in all rearing tanks. P. monodon postlarvae fed probiont-encapsulated Artemia diets (C1 and C2) exhibited the highest survival (94.3 and 82.3%, respectively) and lowest pathogen load (V. harveyi) in hepatopancreas (5.2 × 102 ± 9.0 × 10 and 4.6 × 102 ± 9.0 × 10 CFU g−1, respectively) and muscle (2.0 × 102 ± 6 × 10 and 1.7 × 102 ± 8.6 × 10 CFU g−1, respectively) tissues. The shrimp that were fed the unenriched Artemia (Control; A) showed the lowest survival (26.33%) and highest bacterial load in the hepatopancreas (1.0 × 105 ± 5 × 103 CFU g−1) and muscle (3.6 × 104 ± 6 × 102 CFU g−1). The shrimp fed the herbal product (D)-enriched Artemia also exhibited enhanced survival and reduced V. harveyi load in the tissues tested compared to the control diet (A) group. The results are discussed in terms of developing a quality
larval feed to produce healthy shrimp larvae. 相似文献
20.
为分析感染水霉菌对巨须裂腹鱼脾脏转录组的影响，探索水霉菌感染巨须裂腹鱼的分子机制，实验随机选取生活在同一水域中的5尾健康和5尾感染水霉菌的巨须裂腹鱼，采用PDA琼脂培养基和分子生物学方法对巨须裂腹鱼水霉菌进行分离鉴定，并采用Illumina HiSeqTM 2000 高通量测序平台，对健康和感染水霉菌的巨须裂腹鱼脾脏组织进行转录组测序，并对测序数据进行拼接、注释和差异表达基因分析。结果显示，从巨须裂腹鱼皮肤上分离鉴定得到3株水霉菌；转录组数据显示，健康组和感染水霉病组分别获得46 619 504 条和43 912 876 条数据，与健康组相比，感染水霉菌组共有1 889 个基因发生差异表达，其中1 414 个基因上调，475 个基因下调，随机选取6个差异表达基因进行实时荧光定量PCR (RT-qPCR)验证，验证结果与转录组测序一致。对健康组和感染水霉菌组的差异表达基因进行GO功能富集发现，上调基因主要富集于241个功能中，下调基因主要富集于60个功能中，上述基因主要涉及分子功能类、细胞组分类和生物过程类等生理功能；KEGG通路富集分析发现，差异表达基因主要富集在免疫疾病、内分泌和代谢疾病、病毒感染性疾病、消化系统及排泄系统等。研究表明，感染水霉菌会影响巨须裂腹鱼脾脏组织多种基因的表达量，实验结果为进一步探索巨须裂腹鱼水霉菌的感染机制奠定了基础。 相似文献