Induction, adaptation and recovery of lysosomal integrity in green-lipped mussel Perna viridis

Biomarkers are generally applied to detect pollution in environmental monitoring. Such biological responses should accurately reflect the stress over time in a quantitative manner. As such, the initial and maximum responses induced by stress, as well as adaptation and recovery of these biomarkers, need to be fully understood or else erroneous false-negative or false-positive may be arrived. However, most of the biomarker studies only provided information on initially induced responses under different concentrations of toxicants, while biological adaptation and recovery were poorly known. In this study, the time required for induction, adaptation and recovery of lysosomal integrity in green-lipped mussel Perna viridis upon exposure to benzo[a]pyrene was investigated over a period of 62 days. Maximum induction occurred on day 6 when lysosomal integrity was significantly reduced by 51%, and no further change or adaptation was detected thereafter. When mussels were depurated in clean seawater after 18 days of exposure to benzo[a]pyrene, a gradual recovery was observed, with lysosomal integrity returning to its background level and showing a complete recovery after 20 days of depuration. Lysosomal integrity was significantly correlated with the body burden concentrations of benzo[a]pyrene and condition index of the mussels. The relatively fast induction (6 days) and recovery (20 days) without apparent adaptation suggested that lysosomal integrity in P. viridis can serve as a good biomarker in biomonitoring, as its response is not likely to generate both false-negative and false-positive results.     

Induction, adaptation and recovery of lysosomal integrity in green-lipped mussel Perna viridis

Biomarkers are generally applied to detect pollution in environmental monitoring. Such biological responses should accurately reflect the stress over time in a quantitative manner. As such, the initial and maximum responses induced by stress, as well as adaptation and recovery of these biomarkers, need to be fully understood or else erroneous false-negative or false-positive may be arrived. However, most of the biomarker studies only provided information on initially induced responses under different concentrations of toxicants, while biological adaptation and recovery were poorly known. In this study, the time required for induction, adaptation and recovery of lysosomal integrity in green-lipped mussel Perna viridis upon exposure to benzo[a]pyrene was investigated over a period of 62 days. Maximum induction occurred on day 6 when lysosomal integrity was significantly reduced by 51%, and no further change or adaptation was detected thereafter. When mussels were depurated in clean seawater after 18 days of exposure to benzo[a]pyrene, a gradual recovery was observed, with lysosomal integrity returning to its background level and showing a complete recovery after 20 days of depuration. Lysosomal integrity was significantly correlated with the body burden concentrations of benzo[a]pyrene and condition index of the mussels. The relatively fast induction (6 days) and recovery (20 days) without apparent adaptation suggested that lysosomal integrity in P. viridis can serve as a good biomarker in biomonitoring, as its response is not likely to generate both false-negative and false-positive results.     

不同加载速率下煤岩单轴压缩电荷感应规律研究

利用自主研制的电荷感应仪,建立单轴压缩条件下煤岩电荷感应试验系统.研究了煤、花岗岩、砂岩在不同加载速率下的电荷感应规律.试验结果表明:煤岩电荷感应最大值在应力达到极限强度前出现,且随加载速率增加,电荷最大值比应力极限强度提前出现时间有减短趋势.不同性质煤岩体,电荷感应最大值有较大区别,花岗岩电荷最大值大于煤电荷最大值,煤电荷最大值大于砂岩电荷最大值.当应力较小时,煤岩只产生微量的电荷信号,当应力达到煤岩极限应力的90%左右时产生大量的电荷信号,说明煤岩电荷感应存在应力阈值.因此,电荷感应方法作为预测预报动力灾害是可行的,值得深入研究.     

滇西北地区高山湖泊沃迪错近两百年来环境变化及枝角类群落响应

区域增温和大气氮沉降作用已成为高山湖泊面临的重要环境胁迫,已有高山湖泊生物群落响应的长期模式研究主要集中于藻类而缺乏更高营养级生物(如浮游动物)的系统调查。本研究选择滇西北地区深水型的高山湖泊沃迪错开展沉积物调查,通过多指标分析(总氮、总磷、叶绿素a、氮稳定同位素等)并结合区域气候重建记录,识别近两百年来该湖泊及流域环境的变化历史,进一步利用枝角类群落指标(物种组成、生物量等)定量评价了湖泊生物群落的响应模式与驱动因子。结果表明,湖泊营养水平(如总氮浓度)和初级生产力(叶绿素a浓度等)在过去近两百年总体呈上升趋势。相关分析显示,大气氮沉降和流域外源输入是影响总氮上升的主要因素,同时区域增温和营养盐富集促进了湖泊初级生产力的不断上升。自1960s以来区域升温明显,湖泊营养水平和叶绿素a浓度呈现加速上升的趋势。钻孔中枝角类群落以浮游属种(Daphnia longispina等)为优势种,在1900AD以前D.longispina相对丰度较为稳定(40.83%±8.02%),之后出现下降趋势且在1948—1965年间明显下降,之后再次明显上升并成为主要优势种。排序分析显示,气温、叶绿素a和总...     

Cytological and Physiological Biomarker Responses from Green Mussels, Perna viridis (L.) Transplanted to Contaminated Sites in Hong Kong Coastal Waters

Biological monitoring of marine contaminants often involves the analysis of metals accumulated in mussel tissues. Critiques of this type of monitoring suggest that although relatively good indications of ambient metal concentrations are determined, actual biological harm is not elucidated. Biomarkers, techniques originally developed in medicine, have recently been employed in an attempt to ascertain the health of marine mussels. Biomarkers have been used with good effect in temperate marine pollution monitoring programmes. The use of biomarkers in the toxicological evaluation of tropical and subtropical waters is, however, generally lacking, which is somewhat surprising, as considerable industrialisation and environmental degradation is taking place in the Pacific-rim countries of Southeast Asia. In the present study, a suite of cytological and physiological biomarkers have been measured from mussels (Perna viridis: Mytilidae) transplanted within the coastal waters of Hong Kong. Mussels from contaminated sites, had labile lysosomal membranes, whereas animals from a cleaner reference site had more stable membranes. These cytological responses have been related to metal-induced stress and are simple, cost-effective markers of pollution. Cardiac activity from mussels transplanted to polluted sites was generally similar to that of individuals from the reference site, although a mild tachycardia was associated with one site. The cause of the tachycardia and the use of heart rate in mussels as a biomarker is discussed. Integrated physiological responses to stress, condition indices, revealed mussels from a clean reference site had higher indices than animals from contaminated sites. The relative sensitivities of the cytological and physiological biomarkers deployed in this study and their application to pollution monitoring is discussed.     

Molecular and biochemical biomarkers responses in the mussel Mytilus edulis collected from Southern Brazil coast

Marine ecosystems are typically subjected to a variety of stressors containing complex xenobiotics mixtures. This study aims to evaluate the responses of molecular and biochemical biomarkers in the mussel Mytilus edulis providing data for environmental monitoring programs development in Southern Brazil. Mussels were collected at a polluted site, near Patos Lagoon outfall, and at a control site. Gills, muscle and mantle samples were used for biomarker determinations. Mussels collected at the polluted site significantly increased superoxide dismutase (SOD) and glutathione S-transferase (GST) activities and decreased catalase (CAT) activity. Moreover, an increase in sod1, gstπ and hsp70 mRNA expression was observed. Overall, biochemical and molecular biomarkers responses were observed, but these responses varied depending on the analyzed tissue. These results indicate possible contaminants effects on organisms and the need for effective environmental monitoring programs in this ecosystem.     

神农架大九湖湿地摇蚊群落组成及其环境指示意义

泥炭地是一种水-陆过渡型的湿地生态系统。水体沼泽化过程中,植被等生物群落也随之改变,进而影响泥炭地碳埋藏和发育过程。作为泥炭地生态系统重要的次级生产者,摇蚊群落结构变迁可以为追溯泥炭地环境演变和发育历史提供关键线索。泥炭地在发育过程中将经历各种不同阶段,然而,现有的泥炭地摇蚊群落调查仅限于泥炭生境或泥炭地中开阔水域,较短的生境梯度不足以提供更为全面的摇蚊群落变化信息。神农架大九湖湿地包含泥炭地、湖泊、临时性水体等多种生境,本研究选择该湿地公园作为研究区域,通过采集不同生境的表层沉积物及水体样品,提取底泥亚化石摇蚊头壳,分析大九湖湿地中不同生境下的摇蚊群落结构差异,并对其与环境因子间的关系进行探讨。结果表明:1)大九湖湿地不同生境中摇蚊优势种迥异,泥炭藓藓丘生境中Limnophyes、Psilometriocnemus、Pseudosmittia等半陆生种类为主要优势种,湖泊生境中则以典型静水种Polypedilum nubeculosum-type为主要优势种,而过渡性水域(泥炭地洼地、沟渠及水洼生境)中,静水种与半陆生种共存;2)烧失量、水位埋深和pH是塑造不同生境类型下摇蚊群落结构的显著环境因子,水文条件主要通过改变碳积累过程等其他环境条件间接影响生物群落组成;3)沿水生-半陆生生境梯度,有机质含量和类型均发生显著变化,而摇蚊群落也由静水种、静水/半陆生共存转变为以半陆生种为主,摇蚊群落对生境变化表现出良好的响应过程。本研究揭示了不同生境条件下摇蚊群落的结构差异及影响摇蚊群落结构的潜在因子,为未来基于摇蚊的长时间尺度上泥炭地发育过程分析提供参考依据。     

Integrated use of biomarkers in the mussel Mytilus galloprovincialis for assessing off-shore gas platforms in the Adriatic Sea: results of a two-year biomonitoring program

Despite a large number of gas platforms existing in the Adriatic Sea, which is a semi-enclosed basin characterized by a slow turnover rate and increasing industrial as well as other anthropogenic activities, the effects of these structures on the aquatic ecosystem require further investigation. Since 1998, multidisciplinary studies have been performed by CNR-ISMAR to comply with legislation and to support the development of protocols for the monitoring of offshore activities in the Adriatic Sea. The present study was developed to implement a biomonitoring plan to assess the ecotoxicological effects of the extraction activities of an off-shore gas platform. Biomarkers were evaluated in mussels collected from the platform in relation to physiological stress, DNA damage, cellular damage, oxidative stress and exposure effects. Organic contaminants and trace element bioaccumulation were also assessed in the soft body of the mussels to correlate bioaccumulation of pollutants with biomarker responses. The results indicate an absence of platform-related environmental stress.     

Resolving mechanisms of toxicity while pursuing ecotoxicological relevance?

In this age of modern biology, aquatic toxicological research has pursued mechanisms of action of toxicants. This has provided potential tools for ecotoxicologic investigations. However, problems of biocomplexity and issues at higher levels of biological organization remain a challenge. In the 1980s and 1990s and continuing to a lesser extent today, organisms residing in highly contaminated field sites or exposed in the laboratory to calibrated concentrations of individual compounds were carefully analyzed for their responses to priority pollutants. Correlation of biochemical and structural analyses in cultured cells and tissues, as well as the in vivo exposures led to the production and application of biomarkers of exposure and effect and to our awareness of genotoxicity and its chronic manifestations, such as neoplasms, in wild fishes. To gain acceptance of these findings in the greater environmental toxicology community, “validation of the model” versus other, better-established often rodent models, was necessary and became a major focus. Resultant biomarkers were applied to heavily contaminated and reference field sites as part of effects assessment and with investigations following large-scale disasters such as oil spills or industrial accidents.

Over the past 15 years, in the laboratory, small aquarium fish models such as medaka (Oryzias latipes), zebrafish (Danio rerio), platyfish (Xiphophorus species), fathead minnow (Pimephales promelas), and sheepshead minnow (Cyprinodon variegatus) were increasingly used establishing mechanisms of toxicants. Today, the same organisms provide reliable information at higher levels of biological organization relevant to ecotoxicology. We review studies resolving mechanisms of toxicity and discuss ways to address biocomplexity, mixtures of contaminants, and the need to relate individual level responses to populations and communities.     

Impacts of maintenance dredged material disposal on macrobenthic structure and secondary productivity

The results of a monitoring programme to assess the spatial impacts associated with ongoing dredged material disposal activity at a dispersive, coastal disposal site (southwest UK) are described. Benthic impacts were assessed using benthic community structure and secondary productivity estimates.Analyses of univariate indices (including secondary production) and multivariate community structure revealed differences between stations inside and those outside the disposal site were minimal. Generally, stations within and outside the disposal site were characterised by the same species. Regression models indicated that the variability in biological structure and secondary production was predominantly accounted for by natural variables (e.g., depth, sediment granulometry) with only a small amount of residual variability being due to contaminant variables. Thus, the elevated levels of certain contaminants in the vicinity of the disposal area were not sufficient to result in significant ecological or ecotoxicological changes. We ascribe such findings partly to the dispersive nature of the disposal site.     

Evaluating legacy contaminants and emerging chemicals in marine environments using adverse outcome pathways and biological effects-directed analysis

Natural and synthetic chemicals are essential to our daily lives, food supplies, health care, industries and safe sanitation. At the same time protecting marine ecosystems and seafood resources from the adverse effects of chemical contaminants remains an important issue. Since the 1970s, monitoring of persistent, bioaccumulative and toxic (PBT) chemicals using analytical chemistry has provided important spatial and temporal trend data in three important contexts; relating to human health protection from seafood contamination, addressing threats to marine top predators and finally providing essential evidence to better protect the biodiversity of commercial and non-commercial marine species. A number of regional conventions have led to controls on certain PBT chemicals over several years (termed ‘legacy contaminants’; e.g. cadmium, lindane, polycyclic aromatic hydrocarbons [PAHs] and polychlorinated biphenyls [PCBs]). Analytical chemistry plays a key role in evaluating to what extent such regulatory steps have been effective in leading to reduced emissions of these legacy contaminants into marine environments. In parallel, the application of biomarkers (e.g. DNA adducts, CYP1A-EROD, vitellogenin) and bioassays integrated with analytical chemistry has strengthened the evidence base to support an ecosystem approach to manage marine pollution problems. In recent years, however, the increased sensitivity of analytical chemistry, toxicity alerts and wider environmental awareness has led to a focus on emerging chemical contaminants (defined as chemicals that have been detected in the environment, but which are currently not included in regulatory monitoring programmes and whose fate and biological impacts are poorly understood). It is also known that natural chemicals (e.g. algal biotoxins) may also pose a threat to marine species and seafood quality. Hence complex mixtures of legacy contaminants, emerging chemicals and natural biotoxins in marine ecosystems represent important scientific, economic and health challenges. In order to meet these challenges and pursue cost-effective scientific approaches that can provide evidence necessary to support policy needs (e.g. the European Marine Strategy Framework Directive), it is widely recognised that there is a need to (i) provide marine exposure assessments for priority contaminants using a range of validated models, passive samplers and biomarkers; (ii) integrate chemical monitoring data with biological effects data across spatial and temporal scales (including quality controls); and (iii) strengthen the evidence base to understand the relationship between exposure to complex chemical mixtures, biological and ecological impacts through integrated approaches and molecular data (e.g. genomics, proteomics and metabolomics). Additionally, we support the widely held view that (iv) that rather than increasing the analytical chemistry monitoring of large number of emerging contaminants, it will be important to target analytical chemistry towards key groups of chemicals of concern using effects-directed analysis. It is also important to evaluate to what extent existing biomarkers and bioassays can address various classes of emerging chemicals using the adverse outcome pathway (AOP) approach now being developed by the Organization for Economic Cooperation and Development (OECD) with respect to human toxicology and ecotoxicology.     

A multibiomarker approach in barramundi (Lates calcarifer) to measure exposure to contaminants in estuaries of tropical North Queensland

A suite of biomarkers were measured in barramundi (Lates calcarifer) from five North Queensland estuaries along a perceived pollution gradient. The biomarkers selected were 7-ethoxyresorufin-O-deethylase (EROD), cytochrome P450, fluorescent aromatic compounds (FACs), DNA integrity, RNA:DNA ratio, cholinesterase activity (ChE), condition factor and hepatosomatic index. The resulting database was subjected to uni- and multi-variate analyses in order to assess the most suitable biomarkers to assess pollution in North Queensland estuaries and to classify the environmental quality of the sites. Principal components analysis (PCA) on the biochemical markers revealed that EROD, EROD/P450, DNA damage and to a lesser extent ChE and FACs were found to be responsive to contaminants in the environment while cytochrome P450, condition factor and the hepatosomatic index were found to be less responsive biomarkers. This study has demonstrated the utility of applying a multibiomarker approach in conjunction with traditional analysis of contaminants in providing valuable information in environmental risk assessment.     

Benthic marine pollution around McMurdo Station, Antarctica: A summary of findings

McMurdo Station is the United States Antarctic Research Program's (USARP) centre for scientific and logistical support for most of the continent and is the largest settlement in Antarctica. Over 30 years of human presence with shipping, air-transport, scientific, and municipal activities have resulted in severe but localized nearshore marine benthic pollution. This review will summarize the results of 3 years of research which has documented concentrations of chemical contaminants, changes in community patterns, and the toxicity of sediments to invertebrate species and infaunal communities. The primary contaminants are petroleum hydrocarbons in the sediments of Winter Quarters Bay, the site of shipping activities and a former dumpsite. Total hydrocarbon levels in sediments in Winter Quarters Bay were comparable to the most polluted harbours in temperate latitudes. However, a steep gradient of pollution existed: stations less than 1 km away were nearly pristine. Benthic invertebrate communities changed dramatically along this contamination gradient. The results of both field and laboratory bioassay experiments with contaminated sediments found that biological changes observed in benthic communities around McMurdo Station were most likely caused by hydrocarbons, PCBs, and PCTs. A primary goal of polar pollution research is to determine the response and tolerance of Antarctic biota to physical and chemical anthropogenic disturbance, and the mitigation of negative effects. To this end, future work should include continued monitoring of changes in chemical and community patterns, studies of the physiological responses of Antarctic invertebrates to chemical contamination, and further comparisons of anthropogenic and natural disturbances to benthic communities.     

Geophysical mapping of hyporheic processes controlled by sedimentary architecture within compound bar deposits

Hyporheic exchange influences water quality and controls numerous physical, chemical, and biological processes. Despite its importance, hyporheic exchange and the associated dynamics of solute mixing are often difficult to characterize due to spatial (e.g., sedimentary heterogeneity) and temporal (e.g., river stage fluctuation) variabilities. This study coupled geophysical techniques with physical and chemical sediment analyses to map sedimentary architecture and quantify its influence on hyporheic exchange dynamics within a compound bar deposit in a gravel-dominated river system in southwestern Ohio. Electromagnetic induction (EMI) was used to quantify variability in electrical conductivity within the compound bar. EMI informed locations of electrode placement for time-lapse electrical resistivity imaging (ERI) surveys, which were used to examine changes in electrical resistivity driven by hyporheic exchange. Both geophysical methods revealed a zone of high electrical conductivity in the center of the bar, identified as a fine-grained cross-bar channel fill. The zone acts as a baffle to flow, evidenced by stable electrical conditions measured by time-lapse ERI over the study period. Large changes in electrical resistivity throughout the survey period indicate preferential flowpaths through higher permeability sands and gravels. Grain size analyses confirmed sedimentological interpretations of geophysical data. Loss on ignition and x-ray fluorescence identified zones with higher organic matter content that are locations for potentially enhanced geochemical activity within the cross-bar channel fill. Differences in the physical and geochemical characteristics of cross-bar channel fills play an important role in hyporheic flow dynamics and nutrient processing within riverbed sediments. These findings enhance our understanding of the applications of geophysical methods in mapping riverbed heterogeneity and highlight the importance of accurately representing geomorphologic features and heterogeneity when studying hyporheic exchange processes.     

Spatial processes determine planktonic diatom metacommunity structure of headwater streams

Headwater streams are interesting ecological systems for studies of metacommunities given their high degrees of isolation, unidirectionality of water flow and hierarchical organization in the drainage network. In addition, for passively dispersing aquatic organisms (e.g. planktonic diatoms), connectivity between sites is unidirectional, which implies a higher isolation of headwaters, leading to a greater dispersal limitation between streams. The aim of this study was to evaluate the relative importance of local environmental (i.e., physicochemical variables) and spatial (i.e., distance between sites) components to the planktonic diatom community in 28 headwater streams of the Cerrado, central region of Brazil. Due the fact that each sampling point was located in a different headwater stream, we expected that spatial components (a proxy for diatom dispersal) were more important than local environmental conditions to the structure of the diatom metacommunity. We found 51 morphospecies of diatoms (Bacillariophyceae). The community composition of diatoms showed strong spatial variation, which was also indicated by high beta diversity (mainly contributed by the turnover component). The variation in diatom community composition, species richness, total density, and Local Contribution to Beta Diversity (LCBD) of headwaters streams was explained solely by spatial factors. Based on these results, we inferred that due to dispersal limitation, the diatom communities of these headwaters tend to show distinct species composition, with high species turnover among the local communities. We highlight the importance of headwater streams as reservoirs of biodiversity for a riverine diatom metacommunity.     

Bacterial communities are sensitive indicators of contaminant stress

With many environments worldwide experiencing at least some degree of anthropogenic modification, there is great urgency to identify sensitive indicators of ecosystem stress. Estuarine organisms are particularly vulnerable to anthropogenic contaminants. This study presents bacterial communities as sensitive indicators of contaminant stress. Sediments were collected from multiple sites within inner and outer zones of three heavily modified and three relatively unmodified estuaries. Bacterial communities were censused using Automated Ribosomal Intergenic Spacer Analysis and analysed for a suite of metal and PAH contaminants. Shifts in both bacterial community composition and diversity showed strong associations with sediment contaminant concentrations, particularly with metals. Importantly, these changes are discernable from environmental variation inherent to highly complex estuarine environments. Moreover, variation in bacterial communities within sites was limited. This allowed for differences between sites, zones and estuaries to be explained by variables of interest such as contaminants that vary between, but not within individual sites.     

Long-term divergent tidal flat benthic community recovery following hypoxia-induced mortality

Macrobenthos recovery after hypoxia-induced mass mortality was assessed in an estuarine tidal mudflat during 3 years. During the first 2 years, a Pearson-Rosenberg type of community recovery took place along with the improving bottom water oxygen conditions. After 3 months, spionid polychaetes became superabundant (i.e. opportunistic peak), followed rapidly by a steep decline (i.e. ecotone point). Subsequently, a moderate increase in species richness and a steep increase in biomass, related to the growth of long-lived species occurred (i.e. transition region). Afterwards, however, the recovering community diverged again from the ambient, undisturbed, sediments due to enhanced recruitment success of long-lived species presumably resulting from the lowered interference from bioturbation during early recovery stages in the disturbed plots. Hence, despite early community recovery may be more or less deterministic, lagged divergent community reassembling may occur at the longer-term, thereby contributing to benthos patchiness in areas which are frequently subjected to disturbances.     

The "bioeffect assessment index" (BAI). A concept for the quantification of effects of marine pollution by an integrated biomarker approach

The "bioeffect assessment index" (BAI) is based on the integration of several pathological endpoints measured in the liver of European flounder (Platichthys flesus (L.)) during a long term study of biological effects of pollution in the German Bight. The BAI represents a modification of the "health assessment index" since it includes solely validated biomarkers reflecting toxically induced alterations at different levels of biological organisation in order to quantify the effects of environmental pollution. The concept of the BAI is based on the observation of progressive deleterious effects from early responses to late effects. Specific "key events" were detected, representing progressive stages of functional deterioration. The biomarkers selected from a whole battery of cellular markers for the BAI calculation reflect deleterious effects of various classes of contaminants such as heavy metals, organochlorines, pesticides, PAHs, and therefore reflect general toxicity in an integrative manner. Selected biomarkers were: lysosomal perturbations (reduced membrane stability), storage disorders (lipid accumulation) as early markers for toxic effects of liver cells, and the size of macrophage aggregates and their acid phosphatase activity. The latter two markers are indicative for the modulation of non-specific immune response which represents longer time scale responses after chronic exposure.     

Advancing fracture fragility assessment of pre-Northridge welded column splices

A refined probabilistic assessment of seismic demands and fracture capacity of welded column splice (WCS) connections in welded steel moment resisting frames (WSMRFs) is presented. Seismic demand assessment is performed through cloud-based nonlinear time history analysis (NLTHA) for two case-study structures, i.e., a 4- and a 20- story WSMRFs. Results from NLTHA are used to derive fracture fragility of WCS connections. To this aim, the study investigates (1) optimal ground-motion intensity measures for conditioning probabilistic seismic demand models in terms of global (i.e., maximum inter-story drift ratio) and local (i.e., peak tensile stress in the flange of WCSs) engineering demand parameters of WSMRFs; (2) the effect of ground-motion vertical components on the longitudinal flange stress of WCS connections and their resulting fracture fragility; and (3) the effect of WCS capacity uncertainties on the fracture fragility estimates of those connections. For the latter case, an advanced finite element fracture mechanics-based approach proposed by the authors is employed to capture aleatory and epistemic uncertainties affecting fracture capacities. The focus is on pre-Northridge WCS connections featuring partial joint penetration and brittle materials, making them highly vulnerable to seismic fracture. Fracture fragility results for the case-study structures are compared and discussed, highlighting the importance of the considered issues on fragility estimates, particularly in the case of high-rise structures. Findings from the study contribute shedding some light on the influence of seismic demand and capacity uncertainties on the assessment of fracture fragility of WCS connections. These findings can guide similar performance-based assessment exercises for WSMRFs to inform, for instance, the planning and design of retrofitting strategies for those vulnerable connections.