Transfection is a powerful analytical tool enabling studies of gene products and functions in eukaryotic cells. Successful delivery of genetic material into cells depends on DNA quantity and quality, incubation time and ratio of transfection reagent to DNA, the origin, type and the passage of transfected cells, and the presence or absence of serum in the cell culture. So far a number of transfection methods that use viruses, non-viral particles or physical factors as the nucleic acids carriers have been developed. Among non-viral carriers, the cationic polymers are proposed as the most attractive ones due to the possibility of their chemical structure modification, low toxicity and immunogenicity. In this review the delivery systems as well as physical, biological and chemical methods used for eukaryotic cells transfection are described and discussed.
Determination of environmental flows at the regional scale has been complicated by the fine-scale variability of the needs of aquatic organisms. Therefore, most regional methods are based on observation of hydrological patterns and lack evidence of connection to biological responses. In contrast, biologically sound methods are too detailed and resource-consuming for applications on larger scales. The purpose of this pilot project was to develop an approach that would breach this gap and provide biologically sound rules for environmental flow (eflow) estimation for the region of Poland. The concept was developed using seven river sites, which represent the four of six fish-ecological freshwater body types common in Poland. Each of these types was distinguished based on a specific fish community structure, composed of habitat-use guilds. The environmental significance of the flows for these communities was established with help of the habitat simulation model MesoHABSIM computed for each of the seven sites. The established seasonal environmental flow thresholds were standardized to the watershed area and assigned to the corresponding water body type. With these obtained environmental flow coefficients, a standard-setting formula was created, which is compatible with existing standard-setting approaches while maintaining biological significance. The proposed approach is a first attempt to use habitat suitability models to justify a desktop formula for the regional scale eflow criteria.
Background. Fish-based indices for evaluation of river ecosystem quality have been used since the 1980s, when the Index of Biotic Integrity (IBI) was first introduced. Assessment of the ecological status of rivers, based on fish assemblages is required by the Water Framework Directive. During last 15 years a number of national assessment methods based on fish fauna were developed. The recently designed tool for fish-based assessment of ecological status (EFI+IBI_PL) applied in river monitoring in Poland is presented in this paper. Material and methods. The new European Fish Index EFI+ is a multimetric tool consisting of two specific indices, each with two metrics developed separately for salmonid-and cyprinid-river zones. Those metrics were used in the European intercalibration process to validate national methods. However, the original EFI+ method is not adequate to some lowland river types (physical-factor classification), so it was complemented by a typespecific modification of the Index of Biotic Integrity (IBI_PL). The method was tested on fish data from 493 sites located in 431 surface water bodies sampled in 2011-2012 according to the CEN standard 14011. Results. The EFI+ index was adapted to the specificity of Polish rivers by eliminating some inconsistences of the ecoregion division and problems related to the lack of the Dniester River in the EFI+ software and presented in this paper as EFI+PL. The index of diadromous fish occurrence (D) was also adapted from an original EFI+ method and used as a supplementary assessment tool. Specific IBI metrics were developed for large lowland rivers (with sandy or gravel bottom substrate), organic rivers (flowing through peat areas), and rivers connecting lakes (with the presence or lack of salmonid fish species). A software tool for indices calculation was also developed. The method combination (EFI+IBI_PL) was than tested on a set of 493 monitoring sites sampled in 2011-2012. Both indices classified the highest percentage of sites into moderate ecological state/potential class, but for IBI_ PL this percentage was much higher than for EFI+. Percentage of sites classified to good ecological status or high ecological potential by IBI_PL index were lower than for EFI+. The analysis indicates the consistence of classification for 77% of sites to high/good and below good ecological status by the EFI+PL/IBI_PL method and pressure index. Conclusion. The results of a two-years monitoring program show that the combination of modified EFI+ and IBI methods can be applied as a tool for river ecological status assessment in Poland, however some further method modifications are needed.
Application of instream habitat models such as the Mesohabitat Simulation Model (MesoHABSIM) is becoming increasingly popular. Such models can predict alteration to a river physical habitat caused by hydropower operation or river training. They are a tool for water management planning, especially in terms of requirements of the Water Framework Directive. Therefore, model verification studies, which investigate the accuracy and reliability of the results generated, are essential. An electrofishing survey was conducted in September 2014 on the Stura di Demonte River located in north-western Italy. One hundred and sixteen bullhead—Cottus gobio L.—were captured in 80 pre-exposed area electrofishing (PAE) grids. Observations of bullhead distribution in various habitats were used to validate MesoHABSIM model predictions created with inductive and deductive habitat suitability indices. The inductive statistical models used electrofishing data obtained from multiple mountainous streams, analyzed with logistic regression. The deductive approach was based on conditional habitat suitability criteria (CHSC) derived from expert knowledge and information gathered from the literature about species behaviour and habitat use. The results of model comparison and validation show that although the inductive models are more precise and reflect site- and species-specific characteristics, the CHSC model provides quite similar results. We propose to use inductive models for detailed planning of measures that could potentially impair riverine ecosystems at a local scale, since the CHSC model provides general information about habitat suitability and use of such models is advised in pre-development or generic scale studies. However, the CHSC model can be further calibrated with localized electrofishing data at a lower cost than development of an inductive model.
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