Direct comparison of assessment methods using benthic macroinvertebrates: a contribution to the EU Water Framework Directive intercalibration exercise

The aim of the intercalibration exercise presently performed by the EU is to identify and resolve significant inconsistencies between the ecological quality classifications of EU Member States and the normative definitions of the EU Water Framework Directive. Based on benthic macroinvertebrate data of two European stream types (small siliceous mountain streams and medium-sized lowland streams in Central and Western Europe) we correlated the indices of 10 river quality assessment methods (ASPT, BMWP, DSFI, German Multimetric Index, Saprobic Indices) applied in Austria, Czech Republic, Denmark, Germany, Poland, Slovak Republic, Sweden and United Kingdom. National class boundaries were compared via regression analysis. Assessment methods of the same type (Saprobic Indices, BMWP/ASPT scores) showed best correlation results (R²>0.7). The good quality status boundaries of the national methods deviated up to 25%; thus indicating the necessity to harmonize the national classification schemes. Prerequisites of the presented intercalibration approach are (1) a sufficiently large and consistent dataset representative of the respective common intercalibration types and (2) agreement on common type specific reference conditions.     

Addressing a gap in the Water Framework Directive implementation: Rocky shores assessment based on benthic macroinvertebrates

A gap in the European Water Framework Directive (WFD) is addressed, aiming for the development of an ecological quality status assessment tool based solely on the Biological Quality Element benthic macroinvertebrates from intertidal rocky shores. The proposed Rocky shore Macroinvertebrates Assessment Tool (RMAT) was tested and validated along disturbance gradients (organic enrichment). During the whole process, the response of widely used metrics (e.g. Hurlbert index, Shannon-Wiener index, AZTI’s Marine Biotic Index; Bentix biotic index) and models (i.e., metrics combined) was compared to results provided by the Marine Macroalgae Assessment Tool to the same sampling sites.The RMAT is a multimetric index compliant with the WFD based on the benthic macroinvertebrates community, combining ‘abundance’ (Hurlbert index) and ‘taxonomic composition’ (Bentix index using density and biomass data) metrics. It performed well along anthropogenic disturbance gradients, showing ecological quality increasing from close to far away from the disturbance.The RMAT is a promising tool for rocky shore ecological assessment in the scope of the WFD or other monitoring activities worldwide.     

A comparison of the European Water Framework Directive physical typology and RIVPACS-type models as alternative methods of establishing reference conditions for benthic macroinvertebrates

The EU Water Framework Directive requires European Union Member States to establish ‘type-specific biological reference conditions’ for streams and rivers. Types can be defined by using either a fixed typology (System-A), defined by ecoregions and categories of altitude, catchment area and geology, or by means of an alternative characterisation (System-B) that can use a variety of physical and chemical factors. Several European countries also have existing RIVPACS-type models that give site (rather than stream type) specific predictions of benthic macroinvertebrate communities. In this paper we compare the Water Framework Directive (WFD) System-A physical typology and three existing European multivariate RIVPACS-type models as alternative methods of establishing reference conditions. This work is carried out in Great Britain – using RIVPACS, Sweden – using SWEPAC_SRI and the Czech Republic – using PERLA. We found that in all three countries, all seasons and season combinations, and for all biotic indices tested, RIVPACS-type models were more effective (lower standard deviations of O/E ratios) than models based solely on the WFD System-A variables or null models (based on a single expectation for all sites). We also investigated the explanatory power of whole groups of WFD System-A variables and RIVPACS-type model variables, and the explanatory power of individual variables. We found that variables used in the RIVPACS-type models were often better correlates of macroinvertebrate community variation than the WFD System-A variables. We conclude that this is primarily because while the latter use very broad categories of map-derived variables, the former are based on continuous variables selected for their ecological significance.     

The validation of common European class boundaries for river benthic macroinvertebrates to facilitate the intercalibration process of the Water Framework Directive

The Water Framework Directive (WFD) requires that all the water bodies in Europe be protected and enhanced to achieve Good Ecological Status by 2015. The intercalibration of the biological monitoring results of Member States has to be carried out in relation to classification tasks to guarantee a common understanding of ‘Good Ecological Status’ at a European level. An intercalibration exercise was carried out within the framework of different Geographical Intercalibration Groups (GIGs), each composed of a group of countries that share similar river types and have a discrete geographical range. By means of the intercalibration exercise, common European boundaries were proposed for river invertebrate assessment methods within each GIG. The purpose of this study was to validate the boundaries proposed for the formal intercalibration exercise. A benchmark data set was used, which comprised data, collected in different European countries that satisfied WFD requirements. The data set included a set of reference sites and provided evidence of a high degree of comparability among countries. The STAR Intercalibration Common Metric index (STAR_ICMi) was calculated for benchmark samples and was selected as the index in which national assessment boundaries were expressed. It was applied for the intercalibration exercise in two GIGs. For the STAR_ICMi, the coefficient of variation was also calculated, demonstrating a comparable variability with indices that are based on species level identification. A fixed percentile of reference samples STAR_ICMi values was selected as the boundary between High and Good status. The range from this fixed percentile to the lowest possible value was divided into four equal parts to obtain the remaining class boundaries. The resulting High/Good and Good/Moderate boundaries were compared to the boundaries proposed by GIGs and proved to be in line with those defined in the intercalibration exercise. Even if, for the intercalibration exercise, some procedures to check the Member States (MS) data sets where put into practice, it is the responsibility of each state to guarantee the WFD compliancy of their reference conditions and methods. Accordingly, the process of validation explained in this article, or similar ones, can be an important step forward and demonstrate the comparability of the actual boundaries.     

Testing the European stream typology of the Water Framework Directive for macroinvertebrates

In the EU Water Framework Directive (WFD) a typological framework is defined for assessing the ecological quality of water bodies in the future. The conditions in the Directive impose a strong demand for `new' assessment systems. During the AQEM project an assessment system was developed for European streams using macroinvertebrates. The aim of this study was to test if the typology suggested in the WFD is useful for developing an assessment system for macroinvertebrates in streams. In total 889 streams of 29 stream types were sampled in eight countries all over the major geographical gradients in Europe. These stream types fit the WFD typological demands and fit to the major European geographic regions (ecoregions). The sites included gradients from reference conditions (for the definition see Nijboer et al., 2004) to sites with bad ecological quality. Despite standardisation there were large differences between the participating countries concerning the number of taxa, the number of specimens and the taxonomic resolution. The data, including macroinvertebrates and environmental variables were analysed by using Canonical Correspondence Analysis (CCA). The observed macroinvertebrate distribution largely supported the WFD typological criteria. This means that the major macroinvertebrate distribution patterns in European streams follow climatological and geomorphological conditions and are well distinguished in terms of stream types. Furthermore, it was shown that large scale factors affected the macroinvertebrate distribution even on a very fine scale. Most explanatory variables seemed to be scale independent. Even at a fine scale major factors concerning geology, geomorphology and hydrology added to the species occurrences. Within stream types morphology together with physico-chemistry best explained the macroinvertebrates distribution. In conclusion, the WFD typology is useful for an assessment system for streams using macroinvertebrates. The large scale factors were indeed the variables that explained most of the variation in species composition. But as these factors even strongly act at the scale of stream types, a further refinement is most probably necessary to disentangle typological actors from water quality ones.     

Effects of dredge material placement on benthic macroinvertebrates of the Illinois River

Since the 1930s, dredge material has been removed from the Illinois River and placed along the main channel border in shallow depths to maintain a 2.7 m deep main channel for commercial navigation. Placement of this material changes the sediment composition from primarily silt/clay to primarily sand, and it buries pre-existing benthic invertebrates. During 1997 and 1998, the benthos of an 125 km reach of the middle Illinois River (La Grange Reach) was studied by extracting 1065 Ponar samples from randomly-selected sites which had never received dredge material, received dredge material one year previous, or received dredge material during the current year. Although total numbers of macroinvertebrates collected was lower in 1998 than in 1997, relative abundances of eight targeted taxa were highly similar between years. Chironimidae were most abundant and comprised >66% of all macroinvertebrates collected both years. Differences in densities of Chironomidae, Ephemeroptera, Sphaeriidae, Corbicula fluminea (Muller, 1774), Dreissena polymorpha (Pallas, 1771), Odonata, and Gastropoda among the three classes of dredge material placement were all significant (P< 0.05). For all taxa, densities were highest at sites that had never received dredge material; and, for all taxa except Chironimidae, densities were lowest at sites that received dredge material during the current year. No significant recovery by macroinvertebrates was noticed on dredge areas of this reach after one year (P>0.05). Future operations to maintain a channel for navigation should consider preexisting densities of macroinvertebrate taxa. Because benthic macroinvertebrates are an important component of the food web and shifting sand does not support diverse macroinvertebrate communities, strategic placement of dredge material by avoiding islands or other areas of high macroinvertebrate diversity could improve overall system productivity and biotic integrity of large river-floodplains.     

EU Drinking Water Directive reference methods for enumeration of total coliforms and Escherichia coli compared with alternative methods

AIMS: The reference methods for enumeration of total coliforms and Escherichia coli as stated in the European Drinking Water Directive were compared with alternative methods. METHODS AND RESULTS: Laboratories used the reference method on Lactose TTC agar (LTTC), the Colilert/18 system, Laurysulphate Agar (LSA), Chromocult Coliform Agar and the E. coli Direct Plating (DP) method. They enumerated more total coliforms on LTTC than on LSA. CONCLUSIONS: LTTC is suitable for analysis of very clean water samples only, due to heavy background growth. Colilert/18 is a good alternative but it enumerates a broader group of total coliforms, resulting in higher counts. The DP method appeared to be the best choice for enumeration of E. coli because Colilert/18 produces lower counts and false-negative results. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows the limitations of the EU reference method on LTTC due to lack of selectivity and suggests alternative methods for the enumeration of total coliforms and E. coli.     

The future of fish-based ecological assessment of European rivers: from traditional EU Water Framework Directive compliant methods to eDNA metabarcoding-based approaches

Most of the present EU Water Framework Directive (WFD) compliant fish-based assessment methods of European rivers are multi-metric indices computed from traditional electrofishing (TEF) samples, but this method has known shortcomings, especially in large rivers. The probability of detecting rare species remains limited, which can alter the sensitivity of the indices. In recent years, environmental (e)DNA metabarcoding techniques have progressed sufficiently to allow applications in various ecological domains as well as eDNA-based ecological assessment methods. A review of the 25 current WFD-compliant methods for river fish shows that 81% of the metrics used in these methods are expressed in richness or relative abundance and thus compatible with eDNA samples. However, more than half of the member states' methods include at least one metric related to age or size structure and would have to adapt their current fish index if reliant solely on eDNA-derived information. Most trait-based metrics expressed in richness are higher when computed from eDNA than when computed from TEF samples. Comparable values are obtained only when the TEF sampling effort increases. Depending on the species trait considered, most trait-based metrics expressed in relative abundance are significantly higher for eDNA than for TEF samples or vice versa due to over-estimation of sub-surface species or under-estimation of benthic and rare species by TEF sampling, respectively. An existing predictive fish index, adapted to make it compatible with eDNA data, delivers an ecological assessment comparable with the current approved method for 22 of the 25 sites tested. Its associated uncertainty is lower than that of current fish indices. Recommendations for the development of future fish eDNA-based indices and the associated eDNA water sampling strategy are discussed.     

Effects of benthic macroinvertebrates on the exchange of solutes between sediments and freshwater

Infaunal macrobenthos (tubificid oligochaetes, chironomid larvae, and unionid bivalves) were studied in laboratory microcosms to determine their effects on freshwater sediment diagnesis and the exchange of solutes between sediments and water. Tubificids enhanced the flux of ammonium, bicarbonate, and silica from sediments. After the onset of anoxia, they decreased the flux of iron and phosphate. Chironomids increased the flux of nitrate, bicarbonate, and silica, but did not affect the flux of phosphate. Pore water concentrations were low within the irrigated burrowed zone for chemical species normally high in reduced sediments. Concentration gradients were less steep in the actively irrigated burrow zone, but radial diffusion to and from burrows, increased rates of organic decomposition, and enhanced diatom frustule dissolution rates result in enhanced mass transport from sediments. Data from an experiment with unionid clams demonstrated the presence of radial diffusion gradients. These clams enhanced the chloride and nitrate flux from sediments, decreased the bicarbonate flux, but did not affect the flux of either phosphate or silicate. Although the clams did not actively irrigate their burrows, their effect on sediments was similar to that of chironomids. Comparison of direct and indirect flux estimates showed that both types of estimates could be highly variable. In general, indirect flux estimates were higher than direct flux estimates.     

Towards a multimetric index for the assessment of Dutch streams using benthic macroinvertebrates

This study describes the development of a macroinvertebrate based multimetric index for two stream types, fast and slow running streams, in the Netherlands within the AQEM project. Existing macroinvertebrate data (949 samples) were collected from these stream types from all over the Netherlands. All sites received a ecological quality (post-)classification ranging from 1 (bad status) to 4 (good status) based on biotic and abiotic variables, using a combination of multivariate analysis and expert-judgement. A number of bioassessment metrics was tested for both stream types (fast and slow running streams) to examine their power to discriminate between streams of different ecological quality within each stream type. A metric was selected for inclusion in the final multimetric index when there was no overlap of the 25th and 75th percentile between one (or more) ecological quality class(es). Out of all metrics tested, none could distinguish between all four ecological quality classes without overlap of the 25th and 75th percentile between one or more of the classes. Instead, metrics were selected that could distinguish between one (or more) ecological quality class(es) and all others. Finally, 10 metrics were selected for the assessment of slow running streams and 11 metrics for the assessment of fast running streams. Class boundaries were established, to make the assignment of scores to the individual metrics possible. The class boundaries were set at the 25th and/or 75th percentile of the individual metric values. The individual metrics were combined into a multimetric index. Calibration showed that 67% of the samples from slow running streams and 65% of the samples from fast running streams were classified in accordance to their post-classification. In total, only 8% of the samples differed more than one quality class from the post-classification. The multimetric index was validated with data collected in the Netherlands from 82 sites for the purpose of the AQEM project. Validation showed that 54% of the streams were classified correctly.     

Methodological considerations on the coastal and transitional benthic indicators proposed for the Water Framework Directive

A large data set from the coastal environment of the Mediterranean across benthic enrichment gradients was used to test the performance of various indices as indicators of the seabed environmental conditions. Comparisons involved samples sieved through different mesh sizes, taken in different seasons and taken with different samplers. Most indices were found not to be affected by sieve mesh size, they showed some variations with sampling season and they varied considerably with sampler (sampling size) used, but also they were found to be significantly intercorrelated. Among the indicators tested, the Benthic Quality Index at the family level (BQI-family) was found to be the least sensitive in changes in the sampling configuration; it is highly correlated with all the other indicators and needs less time and taxonomic expertise. Our results indicate that there is a need for standardization of the methodology used during sampling for the Water Framework Directive, since the sampling configuration (and the indicator used) may to a large extend determine the results of the Ecological Status (ES) analysis.     

松花江流域大型底栖动物群落结构与水质评价

【目的】松花江流域是中国最早的工业基地之一,其水生态环境遭到严重破坏,环境保护工作面临巨大挑战。开展松花江流域水质评价及典型生物类群多样性状况调查,可为松花江流域生态系统的保护和修复提供依据。【方法】于2016年7月调查整个松花江流域近岸的大型底栖动物群落组成和测定水质理化指标,开展其水质理化特征评价和生物指数评价,并探讨底栖动物群落分布与水环境因子间的关系。【结果】理化指标评价结果显示,南源松花江水质状况最差,处于中度污染;北源松花江处于轻度污染;梧桐河水质最好,处于良好状态。松花江流域3个河段的底栖动物群落结构存在空间差异性。另外,梧桐河的物种多样性最高,北源松花江次之,南源松花江最低。溶解氧和营养元素K的浓度是驱动底栖动物群落组成发生显著性差异的主要环境因子。生物指数评价结果显示,3个河段水质均处于轻度污染状态。【结论】松花江流域水质处于轻度到中度污染状态。有机污染是松花江流域面临的主要水质环境问题,对松花江流域底栖动物群落结构产生了显著影响。因此,控制有机质的输入是维持松花江流域水生态系统平衡的重要举措之一。     

Design and implementation of rapid assessment approaches for water resource monitoring using benthic macroinvertebrates

Abstract Benthic macroinvertebrates are the group of organisms most widely used for assessment of water resources. Rapid assessment approaches are intended to be efficient and cost effective; savings are found in reduced sampling and more efficient data analysis. Rapid bioassessment programmes have been quickly accepted and now cover most of the United States (US) and equivalent programmes cover all of the United Kingdom (UK). Rapid bioassessment programmes are designed to screen large regions, pinpointing trouble spots worthy of more detailed attention. Fundamental to all rapid bioassessment methods is the classification of streams so that comparisons can be made between reference areas and areas of concern, or test sites with similar characteristics. Both the UK and US approaches assess habitat characteristics. These characteristics are used to predict the fauna expected at a test site in the UK approach; in the US they are used as an aid to classification and interpretation of aquatic faunal data. Habitat assessments in the US are also used to determine whether poor water quality or degraded habitat are stressing the invertebrate communities. This is a major development in approaches to water resource assessment. In the UK, a model developed using multivariate statistics uses a few environmental variables thought to be unaffected by human activities to predict the fauna expected at a test site. The US approaches analyse data using several indices (or metrics) presumed to represent ecological features of interest. These indices have a range of sensitivities to different kinds of stress and must be calibrated for the area of interest. The two approaches have been developed in isolation but may have much to offer each other. Developing programmes are advised to consider both. Future needs include: development of procedures that can be applied to large rivers and to lakes; further refinement of ecological principles underlying metric choice; the inclusion of chemical criteria and toxicity tests to establish thresholds that indicate impairment; and development of criteria indicating the necessity for implementation of quantitative assessment studies.     

Development of a multimetric index based on benthic macroinvertebrates for the assessment of natural wetlands in Southwest Ethiopia

Biotic indices are widely applied for conservation and management of aquatic resources since they allow water resources monitoring agencies to get insight in complex biological data and yield policy relevant information. Despite the worldwide popularity of biotic indices, little information on their use and applicability in Eastern Africa is available. Here, we develop a multimetric index based on macroinvertebrates to assess the ecological condition of natural wetlands in Southwest Ethiopia. Index development was based on a dataset of 222 samples collected during two consecutive years from 63 sites located at eight different wetlands. We used physico-chemical and hydro-morphological variables (land use pattern, habitat alteration, hydrological modification and chemical water quality) to classify sites as reference or degraded. We tested a total of 58 potential metrics representing various aspects of macroinvertebrate assemblages including family richness, composition, tolerance measures and presence and abundance of functional feeding groups. Metrics were selected for the development of a final index based on their sensitivity in discriminating reference from impaired sites, strength of correlation with the anthropogenic disturbance gradient, chemical measurements, and the degree of redundancy. Metrics retained for the final index were:overall family richness, family richness of Ephemeroptera, Odonata and Trichoptera (EOT), and percentage of filterer–collectors. The final index, derived from the sum of three metric scores, was divided into five water quality classes (very bad, bad, moderate, good and very good). Our final multimetric macroinvertebrate index (MMI) distinguished well between reference and impaired wetland sites and showed a significant negative response to a gradient of disturbances (R² = 0.86, p < 0.05). Moreover, it classified a validation dataset accurately with a correctly classified instances of 80% and a Cohen's Kappa value of 0.6. This MMI can be considered as a robust and sensitive tool that can be applied to evaluate the ecological condition of natural wetlands in Ethiopia, where wetland resources are under high pressure as a result of agricultural activities such as grazing and urbanization.     

Atlantic Island freshwater ecosystems: challenges and considerations following the EU Water Framework Directive

The Water Framework Directive or WFD constitutes a major step forward in the protection of the aquatic environment and associated habitats, since it legislates for the characterization of surface water bodies across defined ecoregions and the development of ecological monitoring systems based upon elements of the aquatic biota. The Macaronesian archipelagos include the Azores, Madeira and the Canary Islands. The peripheral situation of the Macaronesian islands has set them apart from many European initiatives concerning the implementation of the WFD, which is biased towards better known continental systems. As a result, they have been included in the same ecoregion as the Iberian Peninsula and the Balearic Islands for management purposes. However, because of their oceanic situation and volcanic origin, the freshwater systems of the Macaronesian islands differ strongly from continental systems in watershed morphology and the composition of the biotic assemblages, which merits separate identification rather than inclusion within the larger Iberic-Macaronesian ecoregion and special attention concerning regional implementation of programmes of measures under the Directive.This paper has not been submitted elsewhere in identical or similar form, nor will it be during the first three months after its submission to Hydrobiologia.     

Monitoring and assessment of water quality in the Haraz River of Iran,using benthic macroinvertebrates indices

The objective of this study was to assess the relationship between river water quality and the distribution of benthic macroinvertebrate communities in the Haraz River in Iran. Using a surber net sampler, benthic macroinvertebrate communities along the stream was sampled in wet and dry seasons of 2015 at nine stations with three replications. The physicochemical water quality parameters were measured in the field by water checker. Hilsenhoff biotic Indices, Shannon Wiener Diversity Indices, Average Score per Taxon (ASPT) Index and Pielou Evenness Index were applied to carry out a biological assessment of water quality. A total of 3781 (spring 769, summer 1092, autumn 1095 and winter 825) benthic macroinvertebrate specimens belonging to 4 orders, 11 classes and 16 families were identified. The lowest number of taxa was recorded in spring while the highest was recorded in autumn. Station 9 had the lowest number of taxa while the highest number of taxa was recorded at station 3. The average values (±SD) of the water quality parameters were temperature 14.75?±?4.38 °C, pH 7.93?±?0.62, water flow 14.11?±?9.04 m³ s^?1, electric conductivity 532.75?±?161.35 μmohs cm^?1, total dissolved solids 296.61?±?76.21 mg L^?1, salinity 0.28?±?0.07 mg L^?1, turbidity 580.77?±?149.92 NTU and dissolved oxygen 8.08?±?0.75 mg L^?1. The assessment of stations 1 to 6 indicated that water quality conditions were suitable. In addition, substantial level of organic pollution was observed in stations 7 and 8. In station 9 water quality was fairly poor, requiring a more favourable management based on the capacity of the self-purification of the Haraz River.     

Effects of land cover type on community structure and functional traits of alpine stream benthic macroinvertebrates

1. In the European Alps, due to the current changes in land use driven by different social and economic factors, grasslands and pastures are being increasingly replaced by forests. Whereas many studies have focused on the impacts of urbanisation and intensive agricultural activities, no study has evaluated the effects of changes among types of natural to semi-natural land cover in the Alps with a focus on lotic environments.
2. Here, combining taxonomic and functional approaches, we show the effects of four different alpine land cover types (rocks, grasslands, coniferous forests, valley bottom pastures) on stream benthic macroinvertebrate communities. Irrespective of elevation, grasslands and pastures exhibited unpredicted similarity in terms of the composition of the macroinvertebrate assemblages and hosted the highest diversity of benthic organisms, whereas in each of the other land cover types, the density and diversity of the faunal assemblages showed distinct and characteristic values.
3. When analysing the functional diversity decomposed to richness, evenness, and divergence components, grasslands and pastures again showed a similar trend, being characterised by high levels of resource exploitation and niche differentiation, with the potential to host additional organisms. However, as expected, differences driven by land cover type and elevation emerged when examining the single functional traits, since elevation played a major role in terms of the distribution of traits conferring resistance and resilience.
4. Our results demonstrate that land cover type is a prominent factor influencing the taxonomic and functional variety of stream benthic macroinvertebrate communities. Moreover, alpine grasslands contained an unexpected diversity of aquatic insects, as previously assessed for other organisms (e.g. plants and snails). Overall, our study highlights the importance of the preservation of the diversity of habitats in the alpine region, with a special focus needed for valuable semi-natural landscapes, such as grasslands and pastures, particularly in a time of increasing intensification and abandonment of lands in the alpine context.

     

Biogeographical patterns of benthic macroinvertebrates in the Southeastern Pacific littoral

Aim To use new collections and taxonomic reviews of the Chilean coastal biota, and studies on biodiversity on a reappraisal of Southeastern Pacific littoral biogeography. Location The temperate coastline of the Southeastern Pacific, extending over 6000 km from northern Peru to the southern tip of Chile. Methods Records of coastal macroinvertebrates were assigned to ten geographic zones along the Chilean coast. The Spearman rank correlation coefficient, ρ, was used to establish the relationship between the total number of species and the number of single records for each zone. Regions and underlying faunal gradient were verified via an unweighted paired group method using arithmetic averages (UPGMA), and ordination non-metric multidimensional scaling (nMDS) analysis based on a disagreement distance matrix of presence/absence data. Abundance down-weighting was preferred to rare species down-weighting, owing to the nature of the records, where a larger number of references for a given taxon did not necessarily indicate a greater abundance or commonness. The dissimilarity matrices between the different zones were used to establish the degree of agreement between the different levels of analysis, using a weighted Spearman rank correlation coefficient ρ_w. Statistical significance was established using a randomization (permutation) test. Main conclusions Analysis of new macroinvertebrate data and taxonomic reviews of the Chilean coastal fauna indicates a Transitional-Temperate Region for the Southeastern Pacific littoral, located between 35°S and 48°S where a gradual mixing and replacement of species negates previous hypotheses on the existence of a marked distributional break at 42°S.     

Active entry of stream benthic macroinvertebrates into the water column

Field experiments investigated the possible active entry of stream benthos into the water column. Over a 1-year period, sediment baskets were suspended for 24 hours in a stream pool so that only swimming or floating organisms, essentially unaided by current, could colonize them. A variety of benthos, including taxa characteristic of riffles, colonized the baskets, with colonization highest in late summer and negligible in winter.A modified drift net towed through the pool was used to quantitatively sample benthic animals actually in the water column. Nighttime tows captured a diverse, abundant fauna and indicated densities substantially higher than invertebrate drift densities reported in the literature. Daytime tows yielded little. Estimated percentages of the benthos in the water column at a given time were generally < 1.0%.These findings suggest that not all invertebrate drift is the result of passive mechanical removal from the substrate by current.     

The efficacy of DNA barcoding in the classification,genetic differentiation,and biodiversity assessment of benthic macroinvertebrates

Macroinvertebrates have been recognized as key ecological indicators of aquatic environment and are the most commonly used approaches for water quality assessment. However, species identification of macroinvertebrates (especially of aquatic insects) proves to be very difficult due to the lack of taxonomic expertise in some regions and can become time‐consuming. In this study, we evaluated the feasibility of DNA barcoding for the classification of benthic macroinvertebrates and investigated the genetic differentiation in seven orders (Insecta: Ephemeroptera, Plecoptera, Trichoptera, Diptera, Hemiptera, Coleoptera, and Odonata) from four large transboundary rivers of northwest China and further explored its potential application to biodiversity assessment. A total of 1,144 COI sequences, belonging to 176 species, 112 genera, and 53 families were obtained and analyzed. The barcoding gap analysis showed that COI gene fragment yielded significant intra‐ and interspecific divergences and obvious barcoding gaps. NJ phylogenetic trees showed that all species group into monophyletic species clusters whether from the same population or not, except two species (Polypedilum. laetum and Polypedilum. bullum). The distance‐based (ABGD) and tree‐based (PTP and MPTP) methods were utilized for grouping specimens into Operational Taxonomic Units (OTUs) and delimiting species. The ABGD, PTP, and MPTP analysis were divided into 177 (p = .0599), 197, and 195 OTUs, respectively. The BIN analysis generated 186 different BINs. Overall, our study showed that DNA barcoding offers an effective framework for macroinvertebrate species identification and sheds new light on the biodiversity assessment of local macroinvertebrates. Also, the construction of DNA barcode reference library of benthic macroinvertebrates in Eurasian transboundary rivers provides a solid backup for bioassessment studies of freshwater habitats using modern high‐throughput technologies in the near future.