Bioassay responses of micro-organisms to sediment elutriates from the St. Lawrence River (Lake St. Louis)

A sediment study, involving both chemical and biological analyses, was carried out in the St. Lawrence River near Montreal (Lake St. Louis). About 60 stations were sampled during 1984–85, and the sediments were analyzed for support variables, heavy metals, and organochlorinated compounds. Subsamples were elutriated using a 1 to 4 sediment/water ratio. The resulting elutriates were analyzed for several chemical variables, while toxicity was measured using the Microtox test, algal ¹⁴C assimilation, and lethality/developmental inhibition in cladocerans, rotifers, and nematodes. The results showed a great variety of responses and sensitivity, and correlations between the tests were non-significant. In terms of toxic responses, the algal and Microtox tests were the most sensitive. Toxic responses could not be explained in simple terms of contaminant concentrations. Therefore, the chemistry of elutriates is not predictive of the toxic potential of contaminated sediments. Biotests can give an insight into the hazard assessment of sediments, but no single test will be sufficient; the use of a battery of standardized biotests, representing different levels of organization/food chain, including representative natural species, is highly recommended.     

化学品植物毒性的评价和鉴别

简述了进行化学品植物毒性评价和鉴别的种类、方法和意义,强调有毒化学品对维管束植物的毒性和严重后果,呼吁加强我国在这方面的研究并尽快建立相应的评价方法和标准,以完善我国化学品生物毒性鉴别和评价方法及标准体系,为保护环境质量,提高人们生活水平,维护我国的生态安全服务。虽然过去由于我国工业化程度较低而没有出现什么问题,但现在必须及时给予重视,防患未然,以免重走西方发达国家的弯路。     

Ecosystem health as measured from the molecular to the community level of organization,with reference to sediment bioassessment

The current recognition that chemical measurements are uncertain indicators of biological consequences of pollution has shifted the emphasis away from assessing environmental chemistry alone toward the inclusion of measurements of the health of organisms. Effects of pollutants begin with the individual, have subsequent repercussions on population level processes, and ramifications for community structure and functions. Pollutants act at a molecular level and the biochemical lesions is the first step in the manifestation of effects. Technologies that operate at the cellular level assist in elucidating toxicity. Higher levels of integration include an organism's capacity for growth. Laboratory bioassays andin situ research can monitor physiological incapacities and assist in predicting population level effects. A yet higher level of organization is that of the ecological community.     

An improved elutriation technique for the bioassessment of sediment contaminants

An improved method is proposed for the preparation of sediment elutriates which permits relatively realistic determination of bioavailable contaminants. It suggests the use of rotary tumbling in a cycle of 3–4 rpm to achieve sediment-water mixing. Experiments were undertaken to evaluate the mixing efficiency of the rotary tumbler as compared to that of the compressed air, wrist-action shaker, and reciprocal shaker methods. Sediment to water ratios of 0 : 1, 1 : 20, 1 : 10, and 1 : 4 were tested over 0.5, 1.0, 24, and 48-h elution periods. Elutriate evaluations were based on chemical, physico-chemical and gravimetric determinations; and also on ¹⁴C-phytoplankton bioassays using Chlorella vulgaris (Beyerinck). Results indicated that rotary tumbling produced the most consistent bioassay-supportable data. It was also the most efficient procedure when used for 1 h with 1 : 4 sediment-water mixtures.     

Biomarker-enhanced assessment of reproductive disorders in Monoporeia affinis exposed to contaminated sediment in the Baltic Sea

Introducing biomarkers into monitoring programs requires understanding of their responses in relation to higher-level biological effects as well as modulating effects of confounding environmental factors. We evaluated relationships between the general toxicity biomarkers (acetylcholinesterase [AChE], lysosomal membrane stability [LMS], oxygen radical absorbance capacity [ORAC]) and reproductive performance (fecundity and embryo aberrations) in the amphipod Monoporeia affinis in the Baltic Sea. To further link biomarker response to contaminant (PCBs, PAHs and metals) levels in the surrounding sediments as well as environmental factors (salinity, bottom depth and total organic carbon in sediments [TOC]), correlation and partial least square regression (PLSR) analyses were applied. The observed contaminants levels were frequently elevated for heavy metals and PAHs, but not PCBs. In the amphipod populations, female ORAC values were positively related to the occurrence of females carrying malformed or membrane-damaged embryos and to the percentage of such embryos in their broods, but also to the fecundity. Female AChE activity was negatively related to the frequency of the membrane-damaged embryos, and positively to the frequency of embryos with arrested development in the broods. Moreover, higher AChE activity and ORAC values in the females occurred at elevated concentrations of metals and PAHs, while there was a negative correlation between embryo ORAC and some PCB congeners. The PLSR models explained over 80% of the variation in the female ORAC and AChE values by variation in contaminant concentrations in combination with environmental variables. Specifically, CB180 and PAM4,9 were identified as negative predictors for ORAC, whereas many PAHs and some metals were positive predictors. The AChE activity was positively related to some metals and negatively to PCBs. In the PLSR models, environmental factors had significant modulating effects, with positive effect of salinity on female ORAC and AChE, and negative effect of TOC on the AChE. The LMS data were less informative, with no apparent relation to any of the contaminants. Linking subcellular responses to the reproduction effects facilitates environmental stress assessment and understanding of the response mechanisms, but also calls for more experimental and field data providing a mechanistic understanding to these linkages.     

Photosynthetic and population growth response of the test alga Selenastrum capricornutum Printz to zinc, cadmium and suspended sediment elutriates

Short-term ¹⁴C-fixation (4 h) Selenastrum capricornutum algal toxicity tests were conducted with Cd (n=8), Zn (n=9) and suspended sediment aqueous elutriates (n=28) and the results were compared to those obtained in a 48 h population growth test. In order to provide more realistic experimental conditions, toxicity tests were carried out in prefiltered nutrient-spiked Lake Geneva water. The population growth inhibition test was significantly more sensitive than the¹⁴ C-fixation test for Cd (median EC50-4h and EC50-48h values of 600 and 118 μg L^-1, respectively) whereas no significant difference was measured for Zn toxicity (median EC50-4h and EC50-48h values of 97 and 96 μg L^-1, respectively). With suspended sediment aqueous elutriates, the relative sensitivity of the two different end points is sample dependent, with ratios of the EC25 for the¹⁴ C-fixation: population growth test ranging from <0.26 to >53.3. Elutriate toxicity shows no apparent relationship between the acute and chronic test, indicating that population growth inhibition cannot be derived directly or predicted from¹⁴ C-fixation. Both tests with their specific advantages and limitations provide valuable complementary information to measure the impact of single toxicants or complex mixtures on aquatic plants. This revised version was published online in June 2006 with corrections to the Cover Date.     

Sediment quality assessment in the delta of rivers Rhine and Meuse based on field observations,bioassays and food chain implications

The quality of sediment was assessed in 46 sites in the delta of the rivers Rhine and Meuse (The Netherlands) by means of physical-chemical analysis, field observations on the macrobenthic community structure, accumulation studies and bioassays using Chironomus riparius, Daphnia magna and Photobacterium phosphoreum. The results of chemical analyses were classified using national criteria for sediment quality. Results of field studies and bioassays were classified using criteria derived from research in reference areas or based on data from literature. Risk assessment was carried out according to the sediment quality triad and by means of multi criteria analysis (MCA). The Triad approach was used to demonstrate causal relations between effects on the macrozoobenthos community structure, effects demonstrated in bioassays and sediment pollution. This was done by making a comparison of sediment contamination levels with toxicity data from literature for the test organisms of the bioassays. Using the MCA method, for each site a numerical value was derived for total environmental risk in the present situation, based on observed effects. In this way, a relative risk ranking of all sites was realized. The MCA values for the present situation were also compared with MCA scores based on estimated risks after remediation in 1995, in order to set priorities for sites where remediation is expected to cause a significant reduction in environmental risk. In most of the 46 sites studied so far, the macrofauna community was poorly developed, judged by a low number of benthic species, low abundances and a high dominance of species regarded as relatively tolerant to chemical pollution. In bioassays high sediment toxicity was demonstrated for 24 sites. Using the sediment quality triad approach, 25 sites were identified as areas where pollution can be held responsible for the effects observed in the field. From a comparison of contaminant concentrations in different types of food with maximum tolerable risk levels, and the application of a bioaccumulation model it was concluded that the sediment pollution also implies high risks for plant-, benthos- or fish-eating birds (secondary poisoning of top predators). In the Nieuwe Merwede highest MCA risk scores were found for shallow parts where highly polluted sediments are found. It is concluded that the sediment quality triad and the MCA provide additional information which can be used to establish priorities for remedial action. Based on an ecotoxicological evaluation of the improved quality of new sediments that will be deposited after removal of the polluted sediments in the Nieuwe Merwede, it is concluded that in this upstream part of the Rhine delta remedial action will be effective.     

Some contributions to the analysis of multivariate data

In this paper, we provide an overview of recently developed methods for the analysis of multivariate data that do not necessarily emanate from a normal universe. Multivariate data occur naturally in the life sciences and in other research fields. When drawing inference, it is generally recommended to take the multivariate nature of the data into account, and not merely analyze each variable separately. Furthermore, it is often of major interest to select an appropriate set of important variables. We present contributions in three different, but closely related, research areas: first, a general approach to the comparison of mean vectors, which allows for profile analysis and tests of dimensionality; second, non‐parametric and parametric methods for the comparison of independent samples of multivariate observations; and third, methods for the situation where the experimental units are observed repeatedly, for example, over time, and the main focus is on analyzing different time profiles when the number p of repeated observations per subject is larger than the number n of subjects.     

Guidance for performing multivariate data analysis of bioprocessing data: Pitfalls and recommendations

Biotech unit operations are often characterized by a large number of inputs (operational parameters) and outputs (performance parameters) along with complex correlations among them. A typical biotech process starts with the vial of the cell bank, ends with the final product, and has anywhere from 15 to 30 such unit operations in series. Besides the above‐mentioned operational parameters, raw material attributes can also impact process performance and product quality as well as interact among each other. Multivariate data analysis (MVDA) offers an effective approach to gather process understanding from such complex datasets. Review of literature suggests that the use of MVDA is rapidly increasing, fuelled by the gradual acceptance of quality by design (QbD) and process analytical technology (PAT) among the regulators and the biotech industry. Implementation of QbD and PAT requires enhanced process and product understanding. In this article, we first discuss the most critical issues that a practitioner needs to be aware of while performing MVDA of bioprocessing data. Next, we present a step by step procedure for performing such analysis. Industrial case studies are used to elucidate the various underlying concepts. With the increasing usage of MVDA, we hope that this article would be a useful resource for present and future practitioners of MVDA. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:967–973, 2014     

Quantification of erosion rates for undisturbed contaminated cohesive sediment cores by image analysis

In order to model the transport of cohesive sediments and associated contaminants accurately, knowledge of erosion rates is indispensable; at present, these are mostly obtained from the literature and usually extended by field data. In contrast to these theoretical approaches we have developed a method to determine erosion rates directly from experiments conducted with undisturbed sediment cores in a laboratory flume. Regularly spaced laser lines are projected onto the sediment surface during the erosion process. To compute the volume loss over a certain period of time, snapshots of the projected lines are taken and the relation between the deformed projected lines and the sediment surface is derived. With this relation and bilinear interpolation, the sediment surface is reconstructed. The volume difference between the reconstructed sediment surfaces corresponding to the snapshots is then calculated. Consequently, volume loss can be determined in mm³ per time with a maximum error of 7.2%. The average error is less than 1%. Combination with the bulk density results in erosion rates in kg m^–1 s^–1. The high sensitivity of the laser lines to sediment surface changes allows the determination of even small erosion rates. Thus, it is possible to determine erosion rates directly as a function of shear stress. Since it is possible to vary the area of calculated sediment-volume loss, scaling effects can be investigated as well.     

Bioavailability and toxicity of sediment-bound lead to a filter-feeder bivalve Crassostrea gigas (Thunberg)

Two different approaches were used to study the bioavailability of sediment-bound lead. In vitro techniques simulating the potential metal desorption under conditions prevailing in the digestive tract were assayed on a contaminated sediment. An experimental model of a food chain was designed to determine the retention of lead in the soft tissues of oysters according to the environmental source of the metal (water or sediment). Neither enzymatic action nor leaching at low pH (both aspects of digestion) induce the release of important lead amounts from particles. Therefore, after 3 weeks of exposure, the retention of lead from the trophic source is lower (1%) compared with direct contamination (5%). Lysosomes are the major intracellular structures responsible for lead storage in the gills, digestive tract and digestive gland. The abundance of lysosomes and their lead amount vary according to the gross concentrations in the soft tissues. The cytopathological data are in agreement with the results about lead accumulation: in oysters exposed to sediment-bound lead, impairments are not so marked as in individuals contaminated directly from water but the same organelles are concerned. Mitochondrial impairments may be related to the effect of lead on cellular respiration processes and changes involving the granular endoplasmic reticulum may have an effect on the level of protein synthesis. Cellular extrusions carrying away numerous lysosomes loaded with lead could account for the balancing of lead incorporation between 2 and 3 weeks of exposure.     

An overall strategy for the testing of chemicals for human hazard and risk assessment under the EU REACH system

In its White Paper, "Strategy for a Future Chemicals Policy," published in 2001, the European Commission (EC) proposed the REACH (Registration, Evaluation and Authorisation of CHemicals) system to deal with both existing and new chemical substances. This system is based on a top-down approach to toxicity testing, in which the degree of toxicity information required is dictated primarily by production volume (tonnage). If testing is to be based on traditional methods, very large numbers of laboratory animals could be needed in response to the REACH system, causing ethical, scientific and logistical problems that would be incompatible with the time-schedule envisaged for testing. The EC has emphasised the need to minimise animal use, but has failed to produce a comprehensive strategy for doing so. The present document provides an overall scheme for predictive toxicity testing, whereby the non-animal methods identified and discussed in a recent and comprehensive ECVAM document, could be used in a tiered approach to provide a rapid and scientifically justified basis for the risk assessment of chemicals for their toxic effects in humans. The scheme starts with a preliminary risk assessment process (involving available information on hazard and exposure), followed by testing, based on physicochemical properties and (Q)SAR approaches. (Q)SAR analyses are used in conjunction with expert system and biokinetic modelling, and information on metabolism and identification of the principal metabolites in humans. The resulting information is then combined with production levels and patterns of use to assess potential human exposure. The nature and extent of any further testing should be based strictly on the need to fill essential information gaps in order to generate adequate risk assessments, and should rely on non-animal methods, as far as possible. The scheme also includes a feedback loop, so that new information is used to improve the predictivity of computational expert systems. Several recommendations are made, the most important of which is that the European Union (EU) should actively promote the improvement and validation of (Q)SAR models and expert systems, and computer-based methods for biokinetic modelling, since these offer the most realistic and most economical solution to the need to test large numbers of chemicals.     

An overall strategy for the testing of chemicals for human hazard and risk assessment under the EU REACH system

In its White Paper, Strategy for a Future Chemicals Policy, published in 2001, the European Commission (EC) proposed the REACH (Registration, Evaluation and Authorisation of CHemicals) system to deal with both existing and new chemical substances. This system is based on a top-down approach to toxicity testing, in which the degree of toxicity information required is dictated primarily by production volume (tonnage). If testing is to be based on traditional methods, very large numbers of laboratory animals could be needed in response to the REACH system, causing ethical, scientific and logistical problems that would be incompatible with the time-schedule envisaged for testing. The EC has emphasised the need to minimise animal use, but has failed to produce a comprehensive strategy for doing so. The present document provides an overall scheme for predictive toxicity testing, whereby the non-animal methods identified and discussed in a recent and comprehensive ECVAM document, could be used in a tiered approach to provide a rapid and scientifically justified basis for the risk assessment of chemicals for their toxic effects in humans. The scheme starts with a preliminary risk assessment process (involving available information on hazard and exposure), followed by testing, based on physicochemical properties and (Q)SAR approaches. (Q)SAR analyses are used in conjunction with expert system and biokinetic modelling, and information on metabolism and identification of the principal metabolites in humans. The resulting information is then combined with production levels and patterns of use to assess potential human exposure. The nature and extent of any further testing should be based strictly on the need to fill essential information gaps in order to generate adequate risk assessments, and should rely on non-animal methods, as far as possible. The scheme also includes a feedback loop, so that new information is used to improve the predictivity of computational expert systems. Several recommendations are made, the most important of which is that the European Union (EU) should actively promote the improvement and validation of (Q)SAR models and expert systems, and computer-based methods for biokinetic modelling, since these offer the most realistic and most economical solution to the need to test large numbers of chemicals.     

Pattern extraction of structural responses of gut microbiota to rotavirus infection via multivariate statistical analysis of clone library data

This study developed a new statistical strategy for analyzing clone library data to observe whether there is a defined pattern in structural responses of gut microbiota to environmental perturbations. A large clone library of genus Bacteroides was constructed with fecal samples for each subject in rotavirus-infected (Group R) and healthy children (Group H). In all, 665 clones of the 12 Group H subjects and 284 clones of the nine Group R subjects were sequenced and classified into 34 operational taxonomic units (OTUs) with a similarity cutoff at 98%. Partial least squares-discriminant analysis was used to observe the change of the Bacteroides spp. composition caused by rotavirus infection and to identify the most relevant species contributing to this shift. It was revealed that H subjects and R subjects were well separated. Bacteroides vulgatus, Bacteroides stercoris and Bacteroides fragilis were identified as the most important discriminating OTUs between two groups. The increased abundance of B. fragilis and the decreased populations of B. vulgatus and B. stercoris in infected guts observed in this study were in agreement with previous culture-based studies. The strategy developed in this work can be used to reveal patterns in structural responses of gut microbiota to environmental perturbations from large-scale 16S rRNA gene-based sequencing data.     

10种杀菌剂对中华蜜蜂的急性毒性测定及风险评估

明确80%代森锰锌可湿性粉剂、50%啶酰菌胺水分散型粒剂、50%腐霉利可湿性粉剂、50%烯酰吗啉可湿性粉剂、100 g/L氰霜唑悬浮剂、20%叶枯唑可湿性粉剂、2%春雷毒素水剂、12.50%腈菌唑乳油、25%丙环唑乳油和15%咪鲜胺微乳剂等10种杀菌剂对中华蜜蜂Apis cerana cerana工蜂的急性毒性,并评价其对中华蜜蜂工蜂的风险性。在室内采用摄入法和点滴法测定10种杀菌剂对中华蜜蜂工蜂的急性经口和接触毒性。结果表明代森锰锌,啶酰菌胺,腐霉利等7种杀菌剂对中华蜜蜂的急性毒性均为低毒;腈菌唑和丙环唑对中华蜜蜂的急性经口毒性和急性接触毒性均为中毒,咪鲜胺对中华蜜蜂的急性经口毒性为高毒,急性接触毒性为中毒;这3种杀菌剂且均属于中等风险性。建议禁止咪鲜胺在蜜源作物开花授粉期施用,慎重选择中等毒性杀菌剂腈菌唑和丙环唑,尽量选择对蜜蜂低毒的杀菌剂,以保护中华蜜蜂的安全。     

DNA extraction for 16S rRNA gene analysis to determine genetic diversity in deep sediment communities

A protocol was devised which permitted the extraction of DNA from deep marine sediments up to 503 m below the sea floor. These sediments have been laid down over the last 3 million years. 16S rRNA gene sequences were amplified from the DNA by the polymerase chain reaction. The details of the successful extraction and polymerase chain reaction methodology varied between samples from different depths. This emphasizes the attention to detail required to allow the diversity of bacteria in these deep sediments to be studied.