14C and delta13C characteristics of organic matter and carbonate in saltmarsh sediments from south west Scotland

The distribution of contaminant radionuclides from the Sellafield nuclear fuel reprocessing plant was used to establish chronologies for three saltmarsh sediment cores from south west Scotland. delta(13)C and (14)C analyses indicated that the cores provided a useful archive record of variations in input of organic matter and carbonate. The results imply that prior to major releases of contaminant (14)C from Sellafield, the (14)C specific activity of organic matter in Irish Sea offshore sediments was about 24 Bq kg(-1) C, while that of the carbonate component was below the limit of detection. These results provide baseline data for modelling the uptake of contaminant (14)C by the Irish Sea sediment system. The study confirmed that small(13)C analyses provide a sensitive means of apportioning the origin of saltmarsh organic matter between C(3) terrigenous plants, C(4) terrigenous plants and suspended particulate marine organic matter. For the <2 mm fraction of sediment, a clear pattern of decreasing marine organic input was observed in response to increasing elevation of the marsh surface as a result of sediment accumulation. Bulk sediment, including detrital vegetation, had a dominant input from terrigenous plants. The combined use of delta(13)C and (14)C data revealed that organic matter in the marine organic component of the <2 mm fraction of contemporary surface sediments of the saltmarshes is dominated by recycled old organic material.     

Effect of light on DIMBOA synthesis in maize leaves as revealed by modified cost-effective extraction method

2,4-Dihydroxy-7-methoxy-1,4-benzoxazine-3-one (DIMBOA) known for its antibiotic properties against many pests and diseases in maize plant was extracted from different varieties. In one of the extraction methods of DIMBOA from maize leaves, Amberlite XAD7, an adsorbent, was used for extracting this and it gave a pure fraction containing DIMBOA. The adsorbent Amberlite XAD, being an expensive material, was uneconomical for processing a large number of samples. Therefore, one of the reported solvent extraction method was modified to make the extraction procedure cost effective without losing the target compounds. Using the modified method, DIMBOA was extracted for different varieties under dark and light condition. Results showed that high concentrations of DIMBOA were extracted from plants grown in dark condition as compared to plants grown in light indicating that light played significant role in inhibition of DIMBOA.     

An examination of the toxic properties of water extracts in the vicinity of an oil sand extraction site

The industrial extraction of oil sands (OS) in northern Alberta, Canada, has raised concerns about the quality of the Athabasca River. The purpose of this study was to examine the toxic properties of various water extracts on Oncorhynchus mykiss trout hepatocytes. The water samples were fractionated on a reverse-phase C(18) cartridge and the levels of light-, medium- and heavy-weight polycyclic aromatic hydrocarbons (PAHs) were determined by fluorescence spectroscopy. Primary cultures of trout hepatocytes were exposed for 48 h at 15 °C to increasing concentrations of the C(18) extract corresponding to 0.02, 0.1, 0.5 and 2.5X concentrations from upstream/downstream sites in the Athabasca River, lake and groundwater samples, OS tailings and interceptor well-water samples. Changes in cell viability, phase I and phase II biotransformation enzymes (cytochrome P4501A and glutathione S-transferase activities), oxidative damage (lipid peroxidation LPO) and genotoxicity (single and double DNA strand breaks) were monitored in post-exposure cells. The water samples decreased cell viability and increased all the above endpoints at thresholds of between 0.02 and 0.1X the water concentration. The most responsive biomarker was DNA damage but it also offered the least discrimination among sites. LPO was higher at sites downstream of the industrial operations compared to upstream sites. A decision tree analysis was performed to formulate a set of rules by which to identify the distinctive properties of each type of water samples. The analysis revealed that OS tailings and interceptor waters were characterized by an increased concentration in light PAHs (>42 μg L(-1)) and this fraction represented more than 85% of the total PAHs. These samples also inhibited GST activity, which could compromise the elimination of genotoxic PAHs present in the system. An analysis of groundwater samples revealed a contamination pattern similar to that for OS tailings. There is a need for more research into specific biomarkers of toxicity from OS tailings compounds such as naphthenic acids, light PAHs among others, which are a characteristic fingerprint of OS extraction activities.     

Assessing the impacts of water abstractions on river ecosystem services: an eco-hydraulic modelling approach

The provision of important river ecosystem services (ES) is dependent on the flow regime. This requires methods to assess the impacts on ES caused by interventions on rivers that affect flow regime, such as water abstractions. This study proposes a method to i) quantify the provision of a set of river ES, ii) simulate the effects of water abstraction alternatives that differ in location and abstracted flow, and iii) assess the impact of water abstraction alternatives on the selected ES. The method is based on river modelling science, and integrates spatially distributed hydrological, hydraulic and habitat models at different spatial and temporal scales. The method is applied to the hydropeaked upper Noce River (Northern Italy), which is regulated by hydropower operations. We selected locally relevant river ES: habitat suitability for the adult marble trout, white-water rafting suitability, hydroelectricity production from run-of-river (RoR) plants. Our results quantify the seasonality of river ES response variables and their intrinsic non-linearity, which explains why the same abstracted flow can produce different effects on trout habitat and rafting suitability depending on the morphology of the abstracted reach. An economic valuation of the examined river ES suggests that incomes from RoR hydropower plants are of comparable magnitude to touristic revenue losses related to the decrease in rafting suitability.     

典型污水处理厂微塑料赋存特征研究

于2021年对镇江市3家典型污水厂水样开展微塑料调查,采用电子显微镜和拉曼光谱仪对水样进行检测分析.结果表明:样品中检出14种微塑料,其中聚乙烯(PE)占比在40％左右,聚丙烯(PP)、聚苯乙烯(PS)、聚对苯二甲酸乙二醇酯(PET)等的占比在5％～40％,尼龙(PA)、聚碳酸酯(PC)、聚甲醛(POM)、聚甲基丙烯酸...     

Simultaneous determination of selected endocrine disrupter compounds in wastewater samples in ultra trace levels using HPLC-ES-MS/MS

An analytical procedure for the simultaneous determination of six selected endocrine disrupter compounds (EDCs: diltiazem, progesterone, benzyl butyl phthalate (BBP), estrone, carbamazepine (Cbz), acetaminophen) was developed by liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-ES-MS/MS). All of the parameters for HPLC and ES-MS/MS systems including mobile phase composition, flow rate, and sample injection volume were optimized to obtain not only the best separation of species interested but also low detection limits. Reverse phase chromatography coupled to ES-MS/MS was used for the separation and detection of EDCs. Formic acid (0.10% ) and 5.0 mM ammonium formate were selected as mobile phase composition in gradient elution. Detection limits for diltiazem, progesterone, BBP, estrone, Cbz, and acetaminophen were found to be 0.13, 0.12, 0.04, 0.13, 0.12, and 0.05 ng/mL, respectively. Influent and effluents from three different wastewater treatment plants located in Ankara, i.e., rotating flat-sheet membrane unit, pilot type flat-sheet membrane unit located at METU Campus and samples from Ankara central wastewater treatment plant were analyzed for their EDCs contents under the optimum conditions.     

Artifact evidence in carbonyl compound sampling using the enclosure technique with cuvette system

Biogenic hydrocarbon emission rates from individual plant species have been estimated experimentally placing small plants or branches in enclosures and measuring the emission rates of the compounds. All-Teflon chambers (cuvettes) where air samples are drawn through tubes packed with adsorbents, have been commonly used for plant enclosure. Sampling of carbonyl compounds emitted from plants has been done using 2,4-dinitrophenylhydrazine coated particles and the derivative carbonyl compounds have been analyzed by HPLC-UV. In this work, the enclosure technique using a cuvette for measurements of carbonyl compound emissions from plants was evaluated. Blank measurements in an empty cuvette have revealed quite different results for experiments performed under laboratory and field conditions. Rigorous univariate statistical analysis of the data obtained indicate that the analytical procedure to determine carbonyl compounds at trace levels in emission samples using the cuvette may lead to positive artifacts during field sampling. This analysis applied to laboratory measurements showed no difference in the results for the cuvette and external lines. Multivariate statistical calculations point out that solar light intensity accounts for the high carbonyl compound levels, especially for acetaldehyde, in the empty cuvette.     

Optimising vegetation monitoring. A case study in A French lowland forest

Biodiversity monitoring surveys are rarely optimised statistically before being initiated. Here, we optimised the monitoring of plants in a temperate forest. The total inventory cost, the number and size of quadrats were optimised to detect a 10% change in species richness over 5 years with α = β = 0.05, using data from ongoing long-term floristic monitoring programs. The procedure showed that the inventory cost would be ca 15% lower using 100-, 200-m² quadrats instead of 300- or 400-m² quadrats. Despite the cost associated with the optimisation (e.g. gathering preliminary data) and the imprecise estimates (due to the typically small sample size of the pilot studies), optimisation would often be a better option than expert opinion when designing a monitoring survey.     

Utilizing pyrosequencing and quantitative PCR to characterize fungal populations among house dust samples

Molecular techniques are an alternative to culturing and counting methods in quantifying indoor fungal contamination. Pyrosequencing offers the possibility of identifying unexpected indoor fungi. In this study, 50 house dust samples were collected from homes in the Yakima Valley, WA. Each sample was analyzed by quantitative PCR (QPCR) for 36 common fungi and by fungal tag-encoded flexible (FLX) amplicon pyrosequencing (fTEFAP) for these and additional fungi. Only 24 of the samples yielded amplified results using fTEFAP but QPCR successfully amplified all 50 samples. Over 450 fungal species were detected by fTEFAP but most were rare. Twenty-two fungi were found by fTEFAP to occur with at least an average of ≥0.5% relative occurrence. Many of these fungi seem to be associated with plants, soil or human skin. Combining fTEFAP and QPCR can enhance studies of fungal contamination in homes.     

Operational problems related to the preparation of the seawater soluble fraction of crude oil

Owing to the importance of dissolution and weathering processes following oil spills, this work focused on the operational (quantitative) aspects related to the dissolution of petroleum-derived products, as well as the influence of solar light on both dissolution and the photoproduction of hydrogen peroxide. Four Brazilian crude oil samples were used to study the transfer process of organic compounds from the crude oil film to the aqueous phase (natural seawater) over a period of up to 45 days. Dissolved organic carbon (DOC), measured by non-dispersive infrared spectroscopy followed by high temperature catalytic combustion, was used to follow the partitioning between the two phases. Aqueous DOC values increased as a function of time (up to 15 days) until equilibrium was reached at concentrations ranging from 5 to 45 mg C L(-1). The final DOC concentration as well as the rate of dissolution depends on the nature of the crude oil. When exposed to sunlight, the dissolution was enhanced by up to 67.3%, and inorganic peroxides were generated in the concentration range from 4.5 up to 8.0 micromol L(-1) after 7.3 h irradiation. These results indicate that there is a need for a standard procedure for the production of the WSF in order to generate a more reliable tool to assess the impact of oil spills on the marine environment.     

The effect of light and iron(II)/iron(III) on the distribution of Tl(I)/Tl(III) in fresh water systems

The distribution of aqueous Tl(I)/Tl(III) as a function of light exposure and solution properties was studied by quantifying the oxidation states after separation with ion chromatography and on-line detection with ICP-MS. Ultraviolet irradiation of aqueous solutions containing 1 microg l(-1) Tl(III) and in equilibrium with the atmosphere increases the reduction rate. In systems with photoreduction of Fe(III)(aq) a quantitative oxidation of Tl(I)(aq) was observed, notably at low pH. The process is reversible, as indicated by formation of Tl(I) when the irradiated systems were kept in the dark. In systems with colloidal silica-stabilised ferrihydrite, UV irradiation also leads to oxidation of Tl(I)(aq), but not quantitatively. It is suggested that adsorption of thallium to the ferrihydrite determines the rate of oxidation. Detectable, but not quantitative, oxidation of Tl(I)(aq) took place when natural water samples with 1 microg l(-1) Tl(I)(aq) were exposed to either sunlight or UV-light. For these samples, the reduction was not quantitative when they were kept in the dark for 24 h. The results suggest that the light dependent iron cycle in fresh water systems strongly influences the redox state of thallium.     

Chemical and biological recovery from acid deposition within the Honnedaga Lake watershed,New York,USA

Honnedaga Lake in the Adirondack region of New York has sustained a heritage brook trout population despite decades of atmospheric acid deposition. Detrimental impacts from acid deposition were observed from 1920 to 1960 with the sequential loss of acid-sensitive fishes, leaving only brook trout extant in the lake. Open-lake trap net catches of brook trout declined for two decades into the late 1970s, when brook trout were considered extirpated from the lake but persisted in tributary refuges. Amendments to the Clean Air Act in 1990 mandated reductions in sulfate and nitrogen oxide emissions. By 2000, brook trout had re-colonized the lake coincident with reductions in surface-water sulfate, nitrate, and inorganic monomeric aluminum. No changes have been observed in surface-water acid-neutralizing capacity (ANC) or calcium concentration. Observed increases in chlorophyll a and decreases in water clarity reflect an increase in phytoplankton abundance. The zooplankton community exhibits low species richness, with a scarcity of acid-sensitive Daphnia and dominance by acid-tolerant copepods. Trap net surveys indicate that relative abundance of adult brook trout population has significantly increased since the 1970s. Brook trout are absent in 65 % of tributaries that are chronically acidified with ANC of <0 μeq/L and toxic aluminum levels (>2 μmol/L). Given the current conditions, a slow recovery of chemistry and biota is expected in Honnedaga Lake and its tributaries. We are exploring the potential to accelerate the recovery of brook trout abundance in Honnedaga Lake through lime applications to chronically and episodically acidified tributaries.     

Research on the relationship between the fractional coverage of the submerged plant Vallisneria spiralis and observed spectral parameters

The present paper discusses the relationship between the coverage fraction of submerged plants and the observed spectral characteristics. The purpose of this paper is to validate a remote sensing technology to monitor the change in the plant composition of a water body. In the current study, the reflectance spectra of the submerged plant Vallisneria spiralis at different fraction coverages of the wetland in Hangzhou Bay were measured. The relationships between the fraction coverage of V. spiralis and simulated Quickbird normalized difference vegetation index (NDVI), red edge, and other spectral characteristic parameters were established. The results showed that the spectral reflectance characteristics of submerged plant V. spiralis were mainly in the visible light (490–650 nm) and near infrared (700–900 nm). The rate of change of the blue band curve and simulated Quickbird NDVI showed a higher correlation with the V. spiralis coverage, so estimation models of the fraction coverage were constructed using these parameters. The estimated fraction coverage of V. spiralis with different models were validated with ground data, and the accuracy of estimation models was assessed. The most suitable estimated fraction coverage of V. spiralis was obtained using the rate of change of the blue band curve and simulated Quickbird NDVI. The present work demonstrated a method to monitor the distribution and dynamical variation of submerged plants at the large scale.     

Measurements of biogenic hydrocarbons and carbonyl compounds emitted by trees from temperate warm Atlantic rainforest, Brazil

This study is part of a three-year project on biogenic volatile organic compound (VOC) emissions from trees of the temperate warm Atlantic rainforest found in the metropolitan area of Sao Paulo City (MASP). No study of VOC emission rates from plant species has been carried out in the temperate warm Atlantic rainforest of Brazil prior to this work. Eleven species were selected (Alchornea sidifolia, Cupania oblongifolia, Cecropia pachystachia, Syagrus romanzoffiana, Casearia sylvestris, Machaerium villosum, Trema micrantha, Croton floribundus, Myrcia rostrata, Solanum erianthum and Ficus insipida) and some of them were studied in urban, sub-urban and forest areas inside the MASP in order to evaluate biogenic VOC composition at sites characterized by different emission sources. Biogenic VOC emissions were determined by placing branches of plants in a dynamic enclosure system, an all-Teflon cuvette, and by sampling the compounds in the air leaving the cuvette. Pre-concentration using adsorbents to retain the VOC, followed by GC-MS after thermal desorption of the sample, was employed to determine the amount of biogenic hydrocarbons. The collection of carbonyl compounds on a 2,4-dinitrophenylhydrazine coated silica followed by HPLC-UV was used to analyze low molecular weight carbonyl compounds. Emission rates of isoprene, alpha-pinene, camphene and limonene ranged from 0.01 to 2.16 microg C h(-1) g(-1) and emission rates of aldehydes (C(2)-C(6)), acrolein, methacrolein and 2-butanone ranged from 1.5 x 10(-2) to 2.3 micro g C h(-1) g (-1). Ambient and leaf temperatures, relative humidity, light intensity, O(3) and NO(x) levels in the local atmosphere were monitored during experiments. It was possible to identify different biogenic VOCs emitted from typical plants of temperate warm Atlantic rainforest. The emission rates were reported as a function of the type of site investigated and were only provided for compounds for which quantification was feasible. Other biogenic compounds were only identified.     

催化剂BiFeO3的制备及其光催化性能研究

采用水热合成法制备了不同煅烧温度下的可见光催化剂BiFeO3粉体,并进行了结构和光催化性能的测试.利用X射线衍射、扫描电镜、傅里叶变换红外光谱和紫外-可见漫反射光谱方法对制备的粉体的晶相结构、表面形貌、光学特性等进行了表征.结果表明,500℃下制备的BiFeO3的晶相最纯、粒径最小、可见光吸收范围广、存在Fe-O键的弯...     

Fungal DNA in hotel rooms in Europe and Asia--associations with latitude, precipitation, building data, room characteristics and hotel ranking

There is little information on the indoor environment in hotels. Analysis of fungal DNA by quantitative PCR (qPCR) is a new method which can detect general and specific sequences. Dust was collected through swab sampling of door frames in 69 hotel rooms in 20 countries in Europe and Asia (2007-2009). Five sequences were detected by qPCR: total fungal DNA, Aspergillus and Penicillium DNA (Asp/Pen DNA), Aspergillus versicolor (A. versicolor DNA), Stachybotrys chartarum (S. chartarum DNA) and Streptomyces spp. (Streptomyces DNA). Associations were analysed by multiple linear regression. Total fungal DNA (GM = 1.08 × 10(8) cell equivalents m(-2); GSD = 6.36) and Asp/Pen DNA (GM = 1.79 × 10(7) cell equivalents m(-2); GSD = 10.12) were detected in all rooms. A. versicolor DNA, S. chartarum DNA and Streptomyces DNA were detected in 84%, 28% and 47% of the samples. In total, 20% of the rooms had observed dampness/mould, and 30% had odour. Low latitude (range 1.5-64.2 degrees) was a predictor of Asp/Pen DNA. Seaside location, lack of mechanical ventilation, and dampness or mould were other predictors of total fungal DNA and Asp/Pen DNA. Hotel ranking (Trip Advisor) or self-rated quality of the interior of the hotel room was a predictor of total fungal DNA, A. versicolor DNA and Streptomyces DNA. Odour was a predictor of S. chartarum DNA. In conclusion, fungal DNA in swab samples from hotel rooms was related to latitude, seaside location, ventilation, visible dampness and indoor mould growth. Hotels in tropical areas may have 10-100 times higher levels of common moulds such as Aspergillus and Penicillium species, as compared to a temperate climate zone.     

Monitoring DNA damage in wood small-reed (Calamagrostis epigejos) plants growing in a sediment reservoir with substrates from uranium mining

For most plant species growing in polluted areas genotoxicity assays are not available. We have studied the possibility of using the alkaline protocol of the Comet assay as a method for detecting induced DNA damage in a grass Calamagrostis epigejos, growing wild in highly polluted areas. To calibrate the Comet assay for C. epigejos, two model mutagens were applied: the monofunctional alkylating agent ethyl methanesulfonate (EMS) and gamma-rays. With increasing concentrations of EMS (0 to 10 mM, 18 h treatment at 26 degrees C) applied on excised leaves, the DNA damage, as expressed by the tail moment (TM) values, increased from 4.7 +/- 0.9 to 60.8 +/- 2.7 microns. After gamma irradiation (0 to 30 Gy) the TM value increased from 4.2 +/- 0.2 to 48.1 +/- 1.7 microns. A 24 to 72 h recovery of leaves after EMS treatment in an EMS-free medium did not result in a significant change in the induced EMS damage. By contrast, a 24 h recovery after gamma-irradiation led to a complete repair of DNA damage measurable by the Comet assay. We have measured the DNA damage in nuclei of leaves of C. epigejos plants growing in the area of a sediment reservoir with substrates from uranium mining, where the ore was exploited through leaching with sulfuric acid. The average specific activity of natural radionuclides measured in the substrate was for 226Ra = 11,818 Bq kg-1, for 232Th = 66 Bq kg-1 and for 40K = 75 Bq kg-1. No significant increase in the DNA damage in plants growing on the sediment substrate above the DNA damage in control plants was detected by the Comet assay.     

Analysis of the changes of vegetation coverage of western Beijing mountainous areas using remote sensing and GIS

Mentougou District acts as a crucial component in the ecological buffer in western Beijing mountainous areas, Beijing, China. Using two Landsat MSS/TM images acquired on July 14, 1979 and July 23, 2005, the vegetation coverage of Mentougou District was calculated based on normalized difference vegetation index and spectral mixture analysis (NDVI-SMA) model. Its temporal and spatial changes were analyzed according to digital elevation model (DEM) image, social and economic data. The results showed that the vegetation coverage decreased from 76.4% in 1979 to 72.7% in 2005. Vegetation degradation was probably the result of human disturbance, such as outspreading of resident areas, and coal and stone mining activities, while vegetation restoration might be contributed by the combined effects of both natural processes and ecological construction effort. Vegetation changes were closely related to topographical characteristics. Plants at high altitude were more stable and less degraded than the plants at low altitude, while the plants on steep slope or northwest aspect were more vulnerable to degradation. During the period of 26 years, landscape appeared to become more fragmental, and ecological quality of the land seemed deteriorated sharply in that highly-covered vegetation area has been decreased by 24%.     

Mycorrhizal colonisation of mountain birch (Betula pubescens ssp. czerepanovii) along three environmental gradients: does life in harsh environments alter plant-fungal relationships?

Environmental stress affects ectomycorrhizal communities (ECM), but it is not known how general the detected ECM responses are. We investigated ECM fungi on roots of mountain birch, Betula pubescens subsp. czerepanovii (Orlova) H?met-Ahti, along three environmental gradients, two natural (altitude, seashore) and one human-induced (pollution), within the Kola Peninsula, NW Russia. Chlorophyll fluorescence of birch leaves indicated no environmental stress even in the conditions that were presumed most stressful in terms of abiotic environment, where the biomass and population density of birches were strongly reduced. Although neither overall ECM colonisation nor root fungal biomass showed stress-related patterns, colonisation by Cenococcum geophilum tended to decrease with abiotic stress. ECM morphotype diversity declined with abiotic stress, and along altitudinal gradient this decline was related to an increase in proportion of morphotypes with high fungal biomass. Polycormic birches had higher ECM colonisation than monocormic birches at high stress sites only. ECM morphotype diversity increased with foliar nitrogen concentration at low stress sites, but not at high stress sites. Birches with higher chlorophyll fluorescence had lower chitin concentration in their roots (indicating lower proportion of fungal structures) at high stress sites only. Our results suggest that at high stress sites (1) mechanical shelter created by polycormic trees may favour ECM fungi and (2) mountain birches maintain lower ECM diversity than at low stress sites.     

Investigations of mercury concentrations in walleye and other fish in the Athabasca River ecosystem with increasing oil sands developments

Recent studies have reported an increasing trend of mercury concentrations in walleye (Sander vitreus) from the Athabasca River, north eastern Alberta (Canada); these studies were based on three years of comparison and attributed the mercury increase to expanding oil sands developments in the region. In order to conduct a more comprehensive analysis of mercury trends in fish, we compiled an extensive database for walleye, lake whitefish (Coregonus clupeaformis), northern pike (Esox lucius) and lake trout (Salvelinus namaycush) using all available data obtained from provincial, federal, and industry-funded monitoring and other programs. Evidence for increasing trends in mercury concentrations were examined for each species by location and year also considering fish weight and length. In the immediate oil sands area of the Athabasca River, mercury concentrations decreased (p < 0.001) in walleye and lake whitefish over 1984-2011. In western Lake Athabasca and its delta, mercury concentrations decreased (p < 0.0001) in northern pike (1981-2009) although no trend was evident for walleye (1981-2005) and lake trout (1978-2009). Mercury concentrations in lake trout from Namur Lake, a small lake west of the oil sands area, were higher in 2007 than 2000 (p < 0.0001); it is difficult to ascribe this increase to an oil sands impact because similar increases in mercury concentrations have been observed in lake trout from similar sized lakes in the Northwest Territories. While mercury emissions rates have increased with oil sands development and the landscape become more disturbed, mercury concentrations remained low in water and sediments in the Athabasca River and its tributaries and similar to concentrations observed outside the development areas and in earlier decades. Our fish database was assembled from a series of studies that differed in study purpose, design, and analytical methods. Future monitoring programs investigating mercury trends in fish should be more rigorous in their design.