A note on sampling chironomids for RNA-based studies of natural populations that retains critical morphological vouchers

The rapid uptake of transcriptomic approaches in freshwater ecology has seen a wealth of data produced concerning the ways in which organisms interact with their environment on a molecular level. Typically, such studies focus either at the community level and so don’t require species identifications, or on laboratory strains of known species identity or natural populations of large, easily identifiable taxa. For chironomids, impediments still exist for applying these technologies to natural populations because they are small-bodied and often require time-consuming secondary sorting of stream material and morphological voucher preparation to confirm species diagnosis. These procedures limit the ability to maintain RNA quantity and quality in such organisms because RNA degrades rapidly and gene expression can be altered rapidly in organisms; thereby limiting the inclusion of such taxa in transcriptomic studies. Here, we demonstrate that these limitations can be overcome and outline an optimised protocol for collecting, sorting and preserving chironomid larvae that enables retention of both morphological vouchers and RNA for subsequent transcriptomics purposes. By ensuring that sorting and voucher preparation are completed within <4 hours after collection and that samples are kept cold at all times, we successfully retained both RNA and morphological vouchers from all specimens. Although not prescriptive in specific methodology, we anticipate that this paper will assist in promoting transcriptomic investigations of the sublethal impact on chironomid gene expression of changes to aquatic environments.

Some statistical aspects of the long-term gill net monitoring programme for pike Esox lucius in Windermere (English Lake District)

For more than 55 years, data have been collected on the population of pike Esox lucius in Windermere, first by the Freshwater Biological Association (FBA) and, since 1989, by the Institute of Freshwater Ecology (IFE) of the NERC Centre for Ecology and Hydrology. The aim of this article is to explore some methodological and statistical issues associated with the precision of pike gill net catches and catch-per-unit-effort (CPUE) data, further to those examined by Bagenal (1972) and especially in the light of the current deployment within the Windermere long-term sampling programme. Specifically, consideration is given to the precision of catch estimates from gill netting, including the effects of sampling different locations, the effectiveness of sampling for distinguishing between years, and the effects of changing fishing effort.

Information science and the scientist. A look at information requirements and the technologies available to scientists in the pursuit of information

A review article looking at the type of information requirements commonly shared by scientists and their use of traditional information services. Areas covered include primary requirements of IFE (Institute of Freshwater Ecology) staff, pure versus applied research, informal and personal sources of information, and traditional library and information services. It goes on to describe how research into information systems and technology may improve the wider accessibility and use of information to the scientific community. Technologies covered include online databases, telecommunications, gateways, expert systems, optical technology and applications of CDROM.

Use of macroinvertebrate communities to detect environmental stress in running waters

This paper presents an account of some current uses of RIVPACS (River Invertebrate Prediction and Classification System), a software package developed by the Institute of Freshwater Ecology (UK). Background information is also given on the unique data-set on which the system is based. Before discussing RIVPACS, we consider the range of environmental stresses encountered in flowing-water systems and some of the ways in which stresses may affect macroinvertebrate communities. The wide application and relevance of the RIVPACS approach was recognised when it was chosen as the biological method for use throughout the UK in the 1990 River Quality Survey (RQS). In the concluding section we list some lessons learnt both from the 1990 survey and from our own testing exercise, and we outline current developments which will lead to a new version of RIVPACS for use in the 1995 RQS.

An introduction to RIVPACS

RIVPACS (River InVertebrate Prediction And Classification System) is a software package developed by the Institute of Freshwater Ecology (IFE). The primary application is to assess the biological quality of rivers within the UK. RIVPACS offers site-specific predictions of the macroinvertebrate fauna to be expected in the absence of major environmental stress. The expected fauna is derived by RIVPACS using a small suite of environmental characteristics. The biological evaluation is then obtained by comparing the fauna observed at the site with the expected fauna. RIVPACS also includes a site classification based on the macroinvertebrate fauna of the component reference sites. New sites, judged by their fauna to be of high biological quality, may be allocated to classification groups within the fixed RIVPACS classification. This has potential for evaluating sites for conservation. In this chapter, the origins and history of the RIVPACS approach are described, including major scientific and operational developments over the life of the project. RIVPACS III is described in detail and predictions at different taxonomic levels are demonstrated. The value of the reference dataset for river management and conservation is examined, and the chapter concludes with a brief consideration of some future challenges.

Genetic differentiation between Atlantic salmon populations in the Windermere catchment

Genetic analysis, using single locus probes for genomic DNA, revealed that the juvenile Atlantic salmon populations in the Rivers Leven, Rothay and Troutbeck were related but genetically distinct. This genetic differentiation is greater than might be expected (by comparison with other salmon populations in the UK) and it is recommended that no action is taken which might promote genetic exchange between the three rivers. Thus, future fisheries management practices should treat the salmon from each site as separate genetic stocks. It is unlikely that any attempts to encourage fish currently spawning in the River Leven (downstream of Windermere) to utilize the upper catchment will be successful. The faster growth rate of juvenile salmon in the River Leven, compared with the River Rothay, probably results from a difference in temperature between the inflowing streams and the main outflow of Windermere. Precocious sexual maturation of some male parr was found in all three populations but the incidence (13-33%) is well within the range reported for other waters. Because of their enhanced growth rate, it is likely that some of the precocious males in the River Leven were 0+ fish. A very high incidence of hybridization (>18%) between Atlantic salmon and brown/sea trout was found in Troutbeck but not in the other rivers. Mitochondrial DNA analysis of these hybrids revealed them to be the product of several, independent cross-fertilizations involving both sexes of both species. The implications of this finding are discussed in relation to the availability of suitable spawning sites in Troutbeck.

Effects of water quality in Bassenthwaite Lake on anglers' catches of salmon and sea-trout in the River Derwent

This is the report on the Effects of Water Quality in the Bassenthwaite Lake on Anglers Catches of Salmon and Sea-trout in the River Derwent April 1993 by the Institute of Freshwater Ecology. An analysis of the catch statistics for salmon and sea-trout in the Rivers Derwent and Cocker was undertaken in relation to available information on the algal water quality in Bassenthwaite Lake to test the hypothesis that poor catch returns were associated with a deterioration of water quality within the lake. Analysis of the catch statistics failed to reveal any correlation between water quality and catch returns for either species of fish and it is concluded that any water deterioration in Bassenthwaite Lake has not caused any major damage to the salmon and sea trout fisheries of the Derwent/Cocker system. This conclusion is supported by the analysis of the Windermere/Leven and Crake system, where no correlation could be found between lake water quality and downstream catches of migratory salmonid fish. However, the possibility still exists and such an effect might be detected by further field work on the macroinvertebrates and on the composition of potential salmonid spawning in the area.

The Cheshire Meres an analysis of data and a prioritisation of sites

This is a report on the Analysis of Data and a Prioritisation of Sites at the Cheshire Meres by the Institute of Freshwater Ecology. The report addresses data collected by the Agency for 24 basin sites in Cheshire. At least two samples were collected from each site, though not simultaneously. Sites were visited in May/June and in November. The determinands are standard and they included: water, temperature, conductivity, pH, DO, fractional white light penetration, TSS, chlorophyll, TP, ortho-phosphate, nitrate-, nitrite-, ammonium and silicate. Though concentrations were often higher than for other lakes in the region, rather exceeding criteria for classification as eutrophic lakes, the results confirmed that the series of lakes is, naturally, highly eutrophic and nothing in the present data differs so far from expectation that is persuasive that the ecosystems are reacting adversely to environmental stress. The data set is review and summarised, site-by-site, in an appendix. The grounds for prioritisation are discussed. Whether or not this preferred prioritised option is adopted, the Agency is recommended to review the way it carries out monitoring. The determinands and the sampling frequency need to be geared to the information that is required.

The Cheshire Meres: a water quality enigma

This is the report on the Cheshire Meres: a Water Quality Enigma by the Institute of Freshwater Ecology from June 1993. The aim of the proposal is to use a well-tested condition-check on key Cheshire Meres, looking at the present nutrient-state and the seasonal structure and relative species-abundances of the phytoplankton. This information will enable the NRA to make some overall appraisal of the advance or otherwise, in trophic condition and its impact upon the perception, use and management of these nationally important sites. The report contains section on the objectives, strategy, timescale, staff and financial of the proposal, plus an appendix with information about the Windermere Profiler and the CV of the Project Leader.

4龄中华鲟垂体的EST分析(英文)

表达序列标签(Expressed sequencetag,EST)是鉴定基因表达规律和发现新基因的一种有效的分子生物学手段。为了能在中华鲟(Acipenser sinensis Gray)中发现与生长和生殖内分泌调控相关的基因,我们构建了中华鲟垂体的SMARTcDNA质粒文库。垂体是调节生长和生殖内分泌的重要器官。在本研究中,通过测序筛选得到了944个EST克隆,将所得EST与GenBank数据库中的序列进行比对,结果表明,802(84.96%)个克隆可以找到同源序列,共代表461个基因,其中含132个已

草鱼呼肠孤病毒上调稀有鮈鲫鳃中TLR3和Mx基因的表达(英文)

鳃暴露在水环境中,增加了对疾病的易感性。为了研究稀有鮈鲫人工感染草鱼呼肠孤病毒过程中鳃部先天性免疫反应机制,我们克隆了抗病毒效应分子Mx基因的部分序列,用适时荧光定量PCR检测双链RNA的模式识别受体(Toll-like receptor3,TLR3)及I型干扰素指示基因Mx的表达。TLR3和Mx基因的表达在注射病毒后12h显著升高(p<0.05),TLR3的表达水平在注射后48h恢复到正常水平(p>0.05),而Mx的高水平表达一直持续到实验结束(p<0.05)。结果表明在GCRV感染中,鳃能发生局部免

七株眼虫基于微卫星DNA指纹图谱的区分和关系分析

尽管以前对眼虫进行过大量的形态发育研究和基于核糖体RNA基因的系统发育分析,但对于株系之间的关系仍然知之甚少．因其形态特征有限并且易变,很难鉴定眼虫的相似种和同种内不同的株．作者利用微卫星DNA指纹图谱,在七株眼虫中扩增了七个微卫星DNA位点,成功扩增的六个微卫星引物都得到了四到八个条带．从微卫星DNA指纹图谱计算得到的相似性系数范围从0．000到0．957．根据相似性系数得到的树状结构,七株眼虫在距离为0．9346处分为三支:E．mutabilis,E．intermedia和E．gracilis．其中,

Influence of dew on biomass and photosystem II activity of cyanobacterial crusts in the Hopq Desert, northwest China

Dew is an important water source for desert organisms in semiarid and arid regions. Both field and laboratory experiments were conducted to investigate the possible roles of dew in growth of biomass and photosynthetic activity within cyanobacterial crust. The cyanobacteria, Microcoleus vaginatus Gom. and Scytonema javanicum (Kutz.) Born et Flah., were begun with stock cultures and sequential mass cultivations, and then the field experiment was performed by inoculating the inocula onto shifting sand for forming cyanobacterial crust during late summer and autumn of 2007 in Hopq Desert, northwest China. Measurements of dew amount and Chlorophyll a content were carried out in order to evaluate the changes in crust biomass following dew. Also, we determined the activity of photosystem II(PSII) within the crust in the laboratory by simulating the desiccation/rehydration process due to dew. Results showed that the average daily dew amount as measured by the cloth-plate method (CPM) was 0.154 mm during fifty-three days and that the crust biomass fluctuated from initial inoculation of 4.3 mu g Chlorophyll a cm(-2) sand to 5.8-7.3 mu g Chlorophyll a cm(-2) crust when dew acted as the sole water source, and reached a peak value of approximately 8.2 mu g Chlorophyll a cm(-2) crust owing to rainfalls. It indicated that there was a highly significant correlation between dew amounts and crust moistures (r = 0.897 or r = 0.882, all P < 0.0001), but not a significant correlation between dew and the biomass (r = 0.246 or r = 0.257, all P > 0.05), and thus concluded that dew might only play a relatively limited role in regulating the crust biomass. Correspondingly, we found that rains significantly facilitated biomass increase of the cyanobacterial crust. Results from the simulative experiment upon rehydration showed that approximately 80% of PSII activity could be achieved within about 50 min after rehydration in the dark and at 5 degrees C, and only about 20% of the activity was light-temperature dependent. This might mean that dew was crucial for cyanobacterial crust to rapidly activate photosynthetic activity during desiccation and rehydration despite low temperatures and weak light before dawn. It also showed in this study that the cyanobacterial crusts could receive and retain more dew than sand, which depended on microclimatic characteristics and soil properties of the crusts. It may be necessary for us to fully understanding the influence of dew on regulating the growth and activity of cyanobacterial crust, and to soundly evaluate the crust's potential application in fighting desertification because of the available water due to dew. (C) 2009 Published by Elsevier Ltd.

Evolutionary Conservation of Dazl Genomic Organization and its Continuous and Dynamic Distribution Throughout Germline Development in Gynogenetic Gibel Carp

To investigate germline development and germ cell specification, we identified a Dazl homolog (CagDazl) from gynogenetic gibel carp (Carassius auratus gibelio). Its cDNA sequence and BAC clone sequence analyses revealed the genomic organization conservation and conserved synteny of the Dazl family members and their neighborhood genes among vertebrates, especially in fish. Moreover, a polyclonal antibody specific to CagDazl was produced and used to examine its expression and distribution throughout germline development at protein level. Firstly, ovary-specific expression pattern of CagDazl was confirmed in adult tissues by RT-PCR and Western blot. In addition, in situ hybridization and immunofluorescence localization demonstrated its specific expression in germ cells, and both its transcript and protein were localized to germ plasm. Then, co-localization of CagDazl and mitochondrial cloud was found, confirming that CagDazl transcript and its protein are germ plasm component and move via METRO pathway during oogenesis. Furthermore, the CagDazl is abundant and continuous throughout germline development and germ cell specification including primordial germ cell (PGC) formation, oogonium differentiation, oocyte development, and embryogenesis, and the dynamic distribution occurs at different development stages. The data suggest that maternal CagDazl might play an important role in gibel carp PGC formation. Therefore, CagDazl is a useful and specific marker for tracing germ plasm and germ cell development in the gynogenetic gibel carp. In addition, in comparison with previous studies in sexual reproduction species, the continuous and dynamic distribution of CagDazl protein in the germ plasm throughout the life cycle seems to have significant implication in sex evolution of vertebrates. J. Exp. Zool. (Mol. Deu. Euol.) 312B:855-871, 2009. (C) 2009 Wiley-Liss, Inc.

Waterborne exposure to PFOS causes disruption of the hypothalamus-pituitary-thyroid axis in zebrafish larvae

Thyroid hormones (THs) play an important role in the normal development and physiological functions in fish. Environmental chemicals may adversely affect thyroid function by disturbing gene transcription. Perfluorooctane sulfonate (PFOS), a persistent compound, is widely distributed in the aquatic environment and wildlife. In the present study, we investigated whether PFOS could disrupt the hypothalamic-pituitary-thyroid (HPT) axis. Zebrafish embryos were exposed to various concentrations of PFOS (0, 100, 200 and 400 mu g L-1) and gene expression patterns were examined 15 d post-fertilization. The expression of several genes in the HIPT system, i.e., corticotropin-releasing factor (CRF), thyroid-stimulating hormone (TSH), sodium/iodide symporter (NIS), thyroglobulin (TG), thyroid peroxidase (TPO), transthyretin (TTR), ioclothyronine deiodinases (Dio1 and Dio2) and thyroid receptor (TR alpha and TR beta), was quantitatively measured using real-time PCR. The gene expression levels of CRF and TSH were significantly up-regulated and down-regulated, respectively, upon exposure to 200 and 400 mu g L-1 PFOS. A significant increase in NIS and Diol gene expression was observed at 200 mu g L-1 PFOS exposure, while TG gene expression was down-regulated at 200 and 400 mu g L-1 PFOS exposure. TTR gene expression was down-regulated in a concentration-dependent manner. Up-regulation and down-regulation of TR alpha and TR beta gene expression, respectively, was observed upon exposure to PFOS. The whole body thyroxine (T-4) content remained unchanged, whereas triiodothyronine (T-3) levels were significantly increased, which could directly reflect disrupted thyroid hormone status after PFOS exposure. The overall results indicated that PFOS exposure could alter gene expression in the HPT axis and that mechanisms of disruption of thyroid status by PFOS could occur at several steps in the synthesis, regulation, and action of thyroid hormones. (C) 2009 Elsevier Ltd. All rights reserved.