Effect of freezing on the length of the penis in Nucella lapillus (L.)

When quantifying imposex in Nucella lapillus, two indices are used: the Vas Deferens Sequence Index (VDSI) and the Relative Penis Size Index (RPSI). Freezing and thawing increase the length of the penis in both male and female Nucella lapillus. In the population studied, this had no significant effect on the RPSI, but was potentially an important source of additional variance in the estimation of the mean penis length.     

Imposex and butyltin concentrations in Bolinus brandaris (Gastropoda: Muricidae) from the northern Tunisian coast

The imposex incidence and butyltin concentration i.e. tributyltin with its di- and mono-substituted metabolites were investigated in the muricid Bolinus brandaris sampled from two sites on the northern Tunisian coast (the Lagoon of Bizerta and the small Gulf of Tunis). Both populations had imposex, with stages of imposex development varying between VDS 1 and VDS 4.3. All imposex indices (imposex frequency (I %), female penis length, female vas deferens length, vas deferens sequence index, relative penis length index, and vas deferens length index) were significantly higher in snails from the Bizerta lagoon. Butyltins were detected in the whole tissues of both sexes from the two sites. TBT levels were higher in gastropods collected from the lagoon of Bizerta (12.65 ±1.48 ng Sn g^???1 dw in female and 15.21 ±1.13 ng Sn g^???1 dw in male) than in individuals from the Gulf of Tunis (10.71 ±1.26 ng Sn g^???1 dw in female and 11.65 ±1.63 ng Sn g^???1 dw in male), corroborating the data of imposex analysis. These results confirmed that B. brandaris could be used as a bioindicator species of butyltin pollution in the studied areas. In addition, this study provided baseline data that could serve for long-term monitoring of TBT pollution in Tunisia, since legislation to reduce the use of TBT-based antifouling paints has not been introduced yet.     

Factors affecting RPSI in imposex monitoring studies using Nucella lapillus (L.) as bioindicator

Nucella lapillus (L.) is a marine gastropod mollusc widely used as a bioindicator of TBT pollution in the North Atlantic coastlines. The species reproductive cycle and the male penis length seasonal/spatial variation were studied at a single site at Aveiro seashore (NW Portugal) between December 2005 and June 2007. The main objective of this work is to assess if the "Relative Penis Size Index" (RPSI)--an important imposex assessment index--varies seasonally and spatially in the same sampling site and how this can affect results obtained in imposex monitoring programmes. Animals able to reproduce were found every month but a seasonal pattern in N. lapillus reproductive cycle was evident. Female gametogenic maturation varied seasonally and a decrease in capsule gland volume and condition factor occurred in late summer/early autumn. The gametogenesis in males did not show a significant seasonal variation as in females but the condition factor, penis length, amount of sperm and prostate volume also diminished in late summer/early autumn. On the other hand, males that were close to egg capsules clusters had larger penises than those far away from clusters. The temporal and spatial male penis length variation introduces a bias on imposex assessment results when using RPSI and the magnitude of the error involved is evaluated for different TBT pollution levels scenarios. We consider that RPSI provides interesting and complementary information that should not be excluded from monitoring programmes, but temporal or spatial comparisons of imposex should be based on other more reliable imposex indices like the VDSI.     

Description and evaluation of imposex in Strombus canarium Linnaeus, 1758 (Gastropoda, Strombidae): a potential bio-indicator of tributyltin pollution

Strombus canarium Linnaeus, 1758 is an important gastropod species within the study area and was traditionally collected for food by the locals. The objective of the present study is to assess the incidence of imposex and its severity in this species. Adult conchs were sampled during their main reproductive period, from October 2005 to January 2006, at Sungai Pulai estuary, Johor Straits, Malaysia. A total of 32.81% of adult females showed imposex characteristics, with varying degrees of severity though. The relative penis size (RPS) index ranged from 1.74 to 33.29 (mean = 13.40 ± 2.27, n?=?21), while the relative penis length (RPL) index ranged from 6.28 to 55.19 (mean = 25.83 ± 3.33, n?=?21). The use of vas deferens sequence (VDS) index was however cannot be applied as the presence of egg groove obscured any vas deferens development in affected females. Sequence of imposex (male penis) development in female conch, from merely a small stump to an advance male penis homologous was therefore carefully analyzed and described, and an alternative imposex classification scheme was proposed. S. canarium can be a good indicator for monitoring of organotin pollution within the study area. However, more studies are needed in order to further develop and test its validity and application, such as its correlation with levels of pollutant within the tissues and the environment, as well as its application on other Strombus species.     

A temporal and spatial monitoring of organotin pollution in a harborside region of Brazil by imposex and ecological quality ratio using <Emphasis Type="Italic">Leucozonia nassa</Emphasis>

This study represents the first use of Leucozonia nassa (Mollusca-Gastropoda) in a program of long-term monitoring. Specimens of L. nassa were collected between 2006 and 2014 and analyzed for the occurrence of imposex in two areas in Southeastern Brazil, one control area and another characterized by the presence of many ports and marinas. Imposex was evaluated in nine sites based on the percentage of females exhibiting imposex (I%), relative penis length index and vas deferens sequence index, classical indices to evaluate tributyltin (TBT) impacts. Moreover, this study uses for the first time in a tropical environment the ecological quality ratio (EQR), an approach to verify the imposex levels as an indicator of the impact by organotin compounds according to the EU Water Framework Directive. The results confirm the occurrence of high levels of imposex in L. nassa with all the indices evaluated. Furthermore, the results also indicate that, despite the international ban, antifouling paints based on TBT are still possibly used in Brazil. In addition, this study shows that L. nassa has the characteristics that make it a suitable bioindicator to monitor TBT pollution, and that the use of the EQR may create important data regarding this problem.     

An innovative statistical approach for analysing non-continuous variables in environmental monitoring: assessing temporal trends of TBT pollution

The current work presents an innovative statistical approach to model ordinal variables in environmental monitoring studies. An ordinal variable has values that can only be compared as "less", "equal" or "greater" and it is not possible to have information about the size of the difference between two particular values. The example of ordinal variable under this study is the vas deferens sequence (VDS) used in imposex (superimposition of male sexual characters onto prosobranch females) field assessment programmes for monitoring tributyltin (TBT) pollution. The statistical methodology presented here is the ordered logit regression model. It assumes that the VDS is an ordinal variable whose values match up a process of imposex development that can be considered continuous in both biological and statistical senses and can be described by a latent non-observable continuous variable. This model was applied to the case study of Nucella lapillus imposex monitoring surveys conducted in the Portuguese coast between 2003 and 2008 to evaluate the temporal evolution of TBT pollution in this country. In order to produce more reliable conclusions, the proposed model includes covariates that may influence the imposex response besides TBT (e.g. the shell size). The model also provides an analysis of the environmental risk associated to TBT pollution by estimating the probability of the occurrence of females with VDS ≥ 2 in each year, according to OSPAR criteria. We consider that the proposed application of this statistical methodology has a great potential in environmental monitoring whenever there is the need to model variables that can only be assessed through an ordinal scale of values.     

The effects of organotin on female gastropods

Female masculinisation (FM) in gastropods (imposex, intersex and ovo-testis) has been documented in > 260 species globally. Biomonitoring studies use FM to indirectly assess the presence of tributyltin (TBT), a once popular biocide in antifouling paint, which has since been regulated. Laboratory studies confirm that FM in gastropods can be induced not only by TBT, but other tin (Sn) compounds, Sn "cocktails", steroids, and other contaminants such as Aroclor 1260. Even though FM in gastropods in the field has been attributed to TBT from boating activity, there is evidence that imposex occurred prior to the advent use of TBT in the 1960s, and for some species imposex occurs naturally. There are roughly 42 species that do not elicit a FM response even if they are exposed to TBT under laboratory conditions, collected sympatrically with other species displaying FM, and/or have TBT in their tissues. A geographic analysis of field studies in four regions indicates that the bulk of studies were conducted in Europe > Asia and Oceania > North America > Latin America. More data are needed regarding FM in Africa, the Middle East, and Latin America since data regarding the masculinisation of female gastropods in these areas are lacking. The most studied gastropod species used in TBT biomonitoring studies is the dogwhelk, Nucella lapillus. This review summarizes the progression of the literature from 1962 to 2009 outlining how the terminology, science, and theories have evolved over time.     

Quality assurance of imposex and intersex measurements in marine snails

Collaboration between the QUASIMEME and BEQUALM projects has proved to be an effective mechanism to further the development of quality assurance procedures for the measurement of the biological effects of tributyltin (imposex and intersex) to meet the requirements of international and national marine monitoring programmes. The data arising from the Laboratory Performance Studies are demonstrating the level of agreement between laboratories, suggesting factors leading to differences between laboratories, and guiding the development of QA procedures.     

Combined use of Nassarius reticulatus imposex and statolith age determination for tracking temporal evolution of TBT pollution in the NW Portuguese continental shelf

Spatial and temporal trends of tributyltin (TBT) pollution on the northern continental shelf of Portugal (5-34 m depth) were assessed using Nassarius reticulatus as a bioindicator. Imposex levels were determined in 2006 and 2010, and integrated with age readings from statoliths for a better chronological analysis of the data. Females affected with imposex were detected in 84% of the sites surveyed in 2006, with levels higher than the Ecological Quality Objective (EcoQO) set by the OSPAR Commission for N. reticulatus (VDSI < 0.3) in approximately half of those sites. In 2010, imposex was only detected in 20% of the surveyed sites and, when detected, the VDSI were lower than the EcoQO, except for one location. The comparison between the current results and the levels previously reported for 2004 and 2005 in the same offshore area evidences a clear decline of imposex in 2010 that confirms the efficacy of Regulation (EC) No. 782/2003 and the IMO AFS-Convention (entered into force in 2003 and 2008, respectively). Moreover, the age of the whelks used in the monitoring is estimated in 3-5 years, indicating that actual TBT water contamination in 2010 may in fact be lower than that extrapolated from the imposex levels. We conclude that the legislation was indeed very effective in reducing TBT pollution in the NW Portuguese continental shelf, not only in terms of the magnitude but also in terms of fast field repercussion.     

Nucella lapillus L. imposex levels after legislation prohibiting TBT antifoulants: temporal trends from 2003 to 2008 along the Portuguese coast

Nucella lapillus (dog whelk) imposex levels were assessed along the mainland Portuguese coast in 2006 and 2008 and were compared with available data from 2003 for the same area. Given that specimen size has been described as a factor inducing variation in some of the imposex assessment indices, and thus resulting in less reliable results, new monitoring and data analysis procedures are described and applied to study change of imposex levels from 2003 to 2008. A significant decline in imposex intensity was observed in the study area during the study period, and the Portuguese coast ecological status (under the terms defined by the OSPAR Commission) notably improved after 2003, confirming the effectiveness of the Regulation (EC) No. 782/2003 in reducing TBT pollution. Nevertheless, N. lapillus populations are still extensively affected by imposex, and fresh TBT inputs were detected in seawater throughout the coast in 2006. These recent inputs are attributed to vessels still carrying TBT antifoulants applied before 2003, as their presence in vessels was only forbidden in 2008. Considering that Regulation (EC) No. 782/2003 is an anticipation of the IMO global ban entered into force in September 2008, a worldwide-scale decrease in TBT pollution can be expected in the near future.     

Imposex measurement in the dogwhelk Nucella lapillus (L.)--temporal aspects of specimen preparation

The measurement of the Relative Penis Size Index (RPSI) in the dogwhelk Nucella lapillus (L.) depends upon reliable observations of the length of the penis (PL). Following removal of soft tissue from the shell, PL increases with time leading to a progressively greater over-estimation of the RPSI. The error is proportionally greater at low RPSI values. To retain comparability with previous data, it is suggested that dogwhelks should be examined as soon as possible after shell crushing.     

A field evaluation of international monitoring guidelines for the biological effects of tributyltin

The Oslo and Paris Commission (OSPAR) Guidelines for monitoring the biological effects of tributyltin compounds (TBT) were evaluated using data collected for preparation of a Celtic Seas Quality Status Report. Two types of survey were undertaken: broad scale, to determine wide impacts in coastal waters; and localised, around representative harbours to establish ranges of effect from recognised input points. This evaluation indicates that results from the broad scale surveys can be used to compare different areas of coastline. Nucella is widespread away from point sources although many individuals show some degree of imposex. Populations are generally not at risk. The localised surveys indicate that, in certain situations, the monitoring objectives can be met in the OSPAR Guidelines (subject to minor amendment). Criteria are identified for the selection of point sources suitable for monitoring under OSPAR Guidelines.     

Characterisation of trace elements and methylmercury in an estuarine sediment reference material, IAEA-405

An estuarine sediment sample, IAEA-405, was recently characterised for trace elements and methylmercury (MeHg) for ultimate use as a marine reference material. The reference values were calculated using data sets from 120 laboratories that participated in a world-wide intercomparison exercise. The data evaluation is highlighted, and includes comparisons of different analytical methods, as well as the distribution of data. Overall laboratory performance for this exercise was encouraging: 86 laboratories (72%) reported data with <25% outliers. There was sufficient acceptable data to establish recommended values for 17 elements and information values for a further 15 elements. In addition, a recommended value was established for methylmercury. The estuarine sediment sample, IAEA-405, can be used as a reference material for quality control in the determination of trace elements and methylmercury in moderately polluted sediments.     

Bias from two analytical laboratories involved in a long-term air monitoring program measuring organic pollutants in the Arctic: a quality assurance/quality control assessment

Initiated in 1992, air monitoring of organic pollutants in the Canadian Arctic provided spatial and temporal trends in support of Canada's participation in the Stockholm Convention of Persistent Organic Pollutants. The specific analytical laboratory charged with this task was changed in 2002 while field sampling protocols remained unchanged. Three rounds of intensive comparison studies were conducted in 2004, 2005, and 2008 to assess data comparability between the two laboratories. Analysis was compared for organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) in standards, blind samples of mixed standards and extracts of real air samples. Good measurement accuracy was achieved for both laboratories when standards were analyzed. Variation of measurement accuracy over time was found for some OCPs and PCBs in standards on a random and non-systematic manner. Relatively low accuracy in analyzing blind samples was likely related to the process of sample purification. Inter-laboratory measurement differences for standards (<30%) and samples (<70%) were generally less than or comparable to those reported in a previous inter-laboratory study with 21 participating laboratories. Regression analysis showed inconsistent data comparability between the two laboratories during the initial stages of the study. These inter-laboratory differences can complicate abilities to discern long-term trends of pollutants in a given sampling site. It is advisable to maintain long-term measurements with minimal changes in sample analysis.     

藻密度手工监测实验室间比对研究

为研究不同实验室藻密度监测数据的可靠性,现场采集了藻密度样品,处理为现场平行样和实验室前处理后平行样2种,分别由5家实验室的6名技术人员开展检测分析。参考相关技术规范要求,结合其他领域相似工作的成功应用案例,采用Z比分数法对分析结果开展藻密度实验室间比对。结果表明,数据的相对偏差均符合技术规范要求;对藻密度进行数据直接比对和对数转换后比对,各实验室对现场平行样和实验室前处理后平行样的检测结果均为合格;分析方法原理导致藻密度手工监测的绝对数值偏差较大,对当前的太湖水华预警工作适用性不高。     

Mercury selection of allozymes in marine gastropods: Prediction and verification in nature revisited

Previous laboratory studies with mercury pollution revealed that the sensitive alleles are (S) for the phosphoglucose isomerase (PGI) locus in both Littorina punctata and L. neritoides and the allele (M+) for the amino peptidase (AP) locus in L. neritoides (Lavie and Nevo, 1986b). The present study tested the frequencies of these alleles at 8 sites along the Israeli Mediterranean shore. We found the frequencies of the sensitive alleles at the mercury polluted site at Akko to be lower than at all other seven sites. Therefore, we concluded that allozyme frequency distributions found in nature are consistent with survivorship patterns found in the laboratory. These results reinforce the suggestion (Nevo et al., 1980) that a monitoring system based on changes in population genetic structure is not only theoretically commendable but practically feasible in principle.     

A proficiency testing scheme for aromatic hydrocarbons in air by the manual thermal desorption-GC method: a comparison of laboratory performance with the uncertainty requirements of the European Union Ambient Air Directive

The Workplace Analysis Scheme for Proficiency (WASP) is a proficiency testing scheme for the analysis of occupational hygiene and environmental air samples and is operated in the UK by the Health and Safety Laboratory. Since 1997, WASP has offered samples of benzene, toluene and m-xylene, at environmental levels on Tenax, and has about 35 laboratories participating, mostly from industry, local government and consultancy organisations in the UK. The results reported cover the first 10 rounds of the environmental analytes (1997-1999) and demonstrate the important role of proficiency testing in assessing the quality of laboratory performance. Estimates are obtained for within-laboratory precision and the total variability at each analyte level. The estimates of within-laboratory precision suggest that laboratories have more difficulty analysing toluene and m-xylene than benzene. Linear relationships for the reproducibility relative standard deviation (RSDT) with loading level are evident for the analytes at occupational levels. At environmental levels, the relationship between loading level and reproducibility is much less well defined. The standard deviation for the proficiency testing assessment for all three analytes at the environmental level is 14%, as derived from the benzene data. Expanded uncertainty estimates (k = 1.96), for the analysis of samples since the scheme started, are obtained from the average total variance, and are 27% for benzene, 39% for toluene and 36% for m-xylene. Although the linear trend of performance against round number was not significant at the 95% level of confidence (p = 0.23 for benzene, p = 0.3 for toluene and p = 0.32 for m-xylene), there was a general improvement in RSDT from 26-34% to about 8-13% 10 rounds later. Currently, for a laboratory to meet one of the data quality objectives in the Ambient Air Directive (indicative measurement of benzene, expanded uncertainty +/- 30% or less), it would have to achieve a level of analytical performance to satisfy the category 1 (best performance) limit of better than +/- 8.8%. In the last proficiency testing round, discussed in this paper, only 58% of laboratories obtained performance scores that indicated that they were able to consistently achieve this level of performance.     

Influence of a collapsed coastal landfill on metal levels in sediments and biota--a portent for the future?

In May 2008 a coastal landslide deposited landfill debris onto the shore near Lyme Regis, UK. Six months later, intertidal sediments and biota from the area were sampled to determine whether the landslip had affected distribution and bioavailability of metals in the area and if there were any biological effects. Highest sediment concentrations for the majority of metals occurred near the landslip zone and in several cases exceeded Threshold Effects or Probable Effects Levels (As, Cu, Hg, Ni, Pb, Zn). The 1 M HCl-extractable fraction of Cd, Pb and Zn in sediments also increased near the landslip. Metal bioaccumulation by intertidal biota showed variability between different species and metals, but there were several instances of increased accumulation near the landslip through increased availability from seawater, sediment and dietary sources. In most cases, metal concentrations in molluscs exceeded Oslo and Paris Commission (OSPAR) background concentrations (BCs) together with background assessment concentrations (BACs) at some sites. Kidney tissues in winkles (Littorina littorea) were measured for evidence of oxidative stress using the Total Oxyradical Scavenging Capacity (TOSC) assay. Responses to peroxynitrite, peroxyl and hydroxyl radicals suggested raised levels of TOSC in animals from the sites close to or east of the landfill waste. There have been very few studies of direct impact of landfills on the marine environment and this study could serve as a practical model for similar events driven by sea level rise.     

用稳健统计法处理方法标准验证实验中方法检出限数据

方法检出限是分析方法基本性能参数之一。根据《环境监测分析方法标准制修订技术导则》(HJ 168—2010)的要求,此参数需要6家以上实验室基于特定样品测试数据的标准偏差得到。目前,标准文本大多采用各家实验室方法检出限中最大值。研究提出用稳健统计法处理方法验证数据,无需识别和删去离群值,可将离群值的影响降到最小。基于4组公开发表数据和2组实测数据,比较了不同方法计算结果。结果表明:参与统计的3组数据29个检测项目中,常规方法确认的方法检出限被判为离群值占27.6%。取最大值存在数值偏高的风险。提出了用曼德尔k检验法识别1组方法检出限数据中的离群值,该方法同样可以用于实验室数据与标准文本中方法检出限符合性判断。     

4-氨基安替比林萃取分光光度法测定水中挥发酚的质量控制指标研究

依据多种浓度、多种样品、多家实验室参与测定而得到的大量监测数据,研究分析了4-氨基安替比林萃取分光光度法测定水中挥发酚的质量控制指标.建议控制值为:当标准样品质量浓度范围为0.04 mg/L～0.2 mg/L时,室内相对标准偏差(RSD)≤5.0％;当标准样品质量浓度范围为0.04 mg/L～1.2 mg/L时,室间相...