Richard Connon - Academia.edu (original) (raw)

Papers by Richard Connon

Research paper thumbnail of An assessment of the toxicity of some pesticides and their metabolites affecting a natural aquatic environment using the Microtox™ system

The conservation and preservation of aquatic ecosystems is of utmost importance due to the high d... more The conservation and preservation of aquatic ecosystems is of utmost importance due to the high diversity and density of species and their complex food network. The evaluation of the potential adverse environmental impact caused by pesticides entering water bodies is an important parameter in aquatic toxicity. The toxicity of the insecticide, Fenitrothion and two of its metabolites, 3-methyl-4 nitrophenol and

Research paper thumbnail of Gene expression responses of threespine stickleback to salinity: implications for salt-sensitive hypertension

Frontiers in Genetics

Despite recent success with genome-wide association studies (GWAS), identifying hypertension (HTN... more Despite recent success with genome-wide association studies (GWAS), identifying hypertension (HTN)-susceptibility loci in the general population remains difficult. Here, we present a novel strategy to address this challenge by studying salinity adaptation in the threespine stickleback, a fish species with diverse salt-handling ecotypes. We acclimated native freshwater (FW) and anadromous saltwater (SW) threespine sticklebacks to fresh, brackish, and sea water for 30 days, and applied RNA sequencing to determine the gene expression in fish kidneys. We identified 1844 salt-responsive genes that were differentially expressed between FW sticklebacks acclimated to different salinities and/or between SW and FW sticklebacks acclimated to full-strength sea water. Significant overlap between stickleback salt-responsive genes and human genes implicated in HTN was detected (P < 10−7, hypergeometric test), suggesting a possible similarity in genetic mechanisms of salt handling between threes...

Research paper thumbnail of Acute and Chronic Invertebrate and Fish Toxicity Testing in the Sacramento-San Joaquin Delta 2008-2009, Final Report, Interagency Ecological Program, Sacramento, CA

Research paper thumbnail of 10-Day survival of Hyalella azteca as a function of water quality parameters

Ecotoxicology and environmental safety, 2015

Estuarine systems are among the most impacted ecosystems due to anthropogenic contaminants; howev... more Estuarine systems are among the most impacted ecosystems due to anthropogenic contaminants; however, they present unique challenges to toxicity testing with regard to varying water quality parameters. The euryhaline amphipod species, Hyalella azteca, is widely used in toxicity testing and well suited for testing estuarine water samples. Nevertheless, the influence of relevant water quality parameters on test endpoints must be quantified in order to efficiently use this species for routine monitoring. Here, we studied the influence of five water quality parameters: electrical conductivity, pH, un-ionized ammonia, dissolved oxygen and temperature, on H. azteca survival in a water column toxicity test. A model was developed to quantify and predict the independent and interacting effects of water quality variables on 10-day survival. The model allows simultaneous assessment of multiple potential predictors recorded during the tests. Data used for modeling came from 1089 tests performed ...

Research paper thumbnail of Effect-based tools for monitoring and predicting the ecotoxicological effects of chemicals in the aquatic environment

Sensors (Basel, Switzerland), 2012

Ecotoxicology faces the challenge of assessing and predicting the effects of an increasing number... more Ecotoxicology faces the challenge of assessing and predicting the effects of an increasing number of chemical stressors on aquatic species and ecosystems. Herein we review currently applied tools in ecological risk assessment, combining information on exposure with expected biological effects or environmental water quality standards; currently applied effect-based tools are presented based on whether exposure occurs in a controlled laboratory environment or in the field. With increasing ecological relevance the reproducibility, specificity and thus suitability for standardisation of methods tends to diminish. We discuss the use of biomarkers in ecotoxicology including ecotoxicogenomics-based endpoints, which are becoming increasingly important for the detection of sublethal effects. Carefully selected sets of biomarkers allow an assessment of exposure to and effects of toxic chemicals, as well as the health status of organisms and, when combined with chemical analysis, identificatio...

Research paper thumbnail of Expression of target and reference genes in Daphnia magna exposed to ibuprofen

BMC genomics, 2006

Transcriptomic techniques are now being applied in ecotoxicology and toxicology to measure the im... more Transcriptomic techniques are now being applied in ecotoxicology and toxicology to measure the impact of stressors and develop understanding of mechanisms of toxicity. Microarray technology in particular offers the potential to measure thousands of gene responses simultaneously. However, it is important that microarrays responses should be validated, at least initially, using real-time quantitative polymerase chain reaction (QPCR). The accurate measurement of target gene expression requires normalisation to an invariant internal control e.g., total RNA or reference genes. Reference genes are preferable, as they control for variation inherent in the cDNA synthesis and PCR. However, reference gene expression can vary between tissues and experimental conditions, which makes it crucial to validate them prior to application. We evaluated 10 candidate reference genes for QPCR in Daphnia magna following a 24 h exposure to the non-steroidal anti-inflammatory drug (NSAID) ibuprofen (IB) at 0...

Research paper thumbnail of Chronic toxicity of ibuprofen to Daphnia magna: Effects on life history traits and population dynamics

Toxicology letters, 2007

The non-steroidal anti-inflammatory drug (NSAID) ibuprofen (IB) is a widely used pharmaceutical t... more The non-steroidal anti-inflammatory drug (NSAID) ibuprofen (IB) is a widely used pharmaceutical that can be found in several freshwater ecosystems. Acute toxicity studies with Daphnia magna suggest that the 48h EC(50) (immobilisation) is 10-100 mgIBl(-1). However, there are currently no chronic IB toxicity data on arthropod populations, and the aquatic life impacts of such analgesic drugs are still undefined. We performed a 14-day exposure of D. magna to IB as a model compound (concentration range: 0, 20, 40 and 80 mgIBl(-1)) measuring chronic effects on life history traits and population performance. Population growth rate was significantly reduced at all IB concentrations, although survival was only affected at 80 mgIBl(-1). Reproduction, however, was affected at lower concentrations of IB (14-day EC(50) of 13.4 mgIBl(-1)), and was completely inhibited at the highest test concentration. The results from this study indicate that the long-term crustacean population consequences of a...

Research paper thumbnail of Is there a relationship between soil and groundwater toxicity?

Environmental geochemistry and health, 2003

Part IIA of the Environmental Protection Act 1990 requires environmental regulators to assess the... more Part IIA of the Environmental Protection Act 1990 requires environmental regulators to assess the risk of contaminants leaching from soils into groundwater (DETR, 1999). This newly introduced legislation assumes a link between soil and groundwater chemistry, in which rainwater leaches contaminants from soil into the saturated zone. As the toxicity of both groundwater and overlying soils is dependent upon the chemicals present, their partitioning and their bioavailability, similar patterns of soil, leachates and groundwater toxicity should be observed at contaminated sites. Soil and groundwater samples were collected from different contaminated land sites in an urban area, and used to determine relationships between soil chemistry and toxicity, mobility of contaminants, and groundwater chemistry and toxicity. Soils were leached using water to mimic rainfall, and both the soils and leachates tested using bioassays. Soil bioassays were carried out using Eisenia fetida, whilst groundwat...

Research paper thumbnail of Durieux et al CYP1A 2012 v def

The striped bass (Morone saxatilis) supports a valuable recreational fishery and is among the mos... more The striped bass (Morone saxatilis) supports a valuable recreational fishery and is among the most important piscivorous fish of the San Francisco Estuary. This species has suffered a significant decline in numbers over the past decades, and there is indication that contaminants are important contributors. Polycyclic aromatic hydrocarbons (PAHs) and polyhalogenated aromatic hydrocarbons (PHAHs) including PCBs and dioxins are widespread in the estuary, they typically bioaccumulate through trophic levels, reaching highest levels in top predators and are known to affect the fish health and development. The aim of this study was to

Research paper thumbnail of Haem peroxidase activity in Daphnia magna: a biomarker for sub-lethal toxicity assessments of kerosene-contaminated groundwater

Ecotoxicology (London, England), 2003

A novel biomarker was developed in Daphnia magna to detect organic pollution in groundwater. The ... more A novel biomarker was developed in Daphnia magna to detect organic pollution in groundwater. The haem peroxidase assay, which is an indirect means of measuring oxidase activity, was particularly sensitive to kerosene contamination. Exposure to sub-lethal concentrations of kerosene-contaminated groundwater resulted in a haem peroxidase activity increase by dose with a two-fold activity peak at 25%. Reproduction in D. magna remained unimpaired when exposed to concentrations below 25% for 21 days, and a decline in fecundity was only observed at concentrations above the peak in enzyme activity. The measurement of haem peroxidase activity in D. magna detected sublethal effects of kerosene in just 24 h, whilst offering information on the health status of the organisms. The biomarker may be useful in determining concentrations above which detrimental effects would occur from long-term exposure for fuel hydrocarbons. Moreover, this novel assay detects exposure to chemicals in samples that w...

Research paper thumbnail of Expression and function of ryanodine receptor related pathways in PCB tolerant Atlantic killifish (Fundulus heteroclitus) from New Bedford Harbor, MA, USA

Aquatic Toxicology, 2015

Atlantic killifish (Fundulus heteroclitus) thrive in New Bedford Harbor (NBH), MA, highly contami... more Atlantic killifish (Fundulus heteroclitus) thrive in New Bedford Harbor (NBH), MA, highly contaminated with polychlorinated biphenyls (PCBs). Resident killifish have evolved tolerance to dioxin-like (DL) PCBs, whose toxic effects through the aryl hydrocarbon receptor (AhR) are well studied. In NBH, non-dioxin like PCBs (NDL PCBs), which lack activity toward the AhR, vastly exceed levels of DL congeners yet how killifish counter NDL toxic effects has not been explored. In mammals and fish, NDL PCBs are potent activators of ryanodine receptors (RyR), Ca 2+ release channels necessary for a vast array of physiological processes. In the current study we compared the expression and function of RyR related pathways in NBH killifish with killifish from the reference site at Scorton Creek (SC, MA). Relative to the SC fish, adults from NBH displayed increased levels of skeletal muscle RyR1 protein, and increased levels of FK506-binding protein 12 kDa (FKBP12) an accessory protein essential for NDL PCB-triggered changes in RyR channel function. In accordance with increased RyR1 levels, NBH killifish displayed increased maximal ligand binding, increased maximal response to Ca 2+ activation and increased maximal response to activation by the NDL PCB congener PCB 95. Compared to SC, NBH embryos and larvae had increased levels of mtor and ryr2 transcripts at multiple stages of development, and generations, while levels of serca2 were decreased at 9 days post-fertilization in the F1 and F2 generations. These findings suggest that there are compensatory and heritable changes in RyR mediated Ca 2+ signaling proteins or potential signaling partners in NBH killifish.

Research paper thumbnail of From'omics to otoliths: Using responses to endocrine disrupting compounds at multiple biological scales to predict population dynamics

ABSTRACT Background/Question/Methods Endocrine disrupting chemicals (EDCs) interfere with endogen... more ABSTRACT Background/Question/Methods Endocrine disrupting chemicals (EDCs) interfere with endogenous hormones and cause physiological abnormalities in fishes. Both theory and empirical data confirm that EDCs can also cause declines in fish populations. However, few studies have attempted to link complex environmental EDC mixtures with responses at multiple tiers of the biological hierarchy, including population-level effects. Additionally, the use of environmentally relevant fishes as model species from regions with demonstrated fish population decline is lacking. To this end, we undertook a four-tiered investigation into estrogenic and androgenic EDC effects in Menidia audens (Mississippi silverside), an atherinid distributed throughout the impacted San Francisco Bay (SFB) estuary. Our main objective was to integrate observations at each biological scale with the goal of determining whether the reproductive health of Menidia populations is being negatively impacted by sites receiving either urban run-off and treated wastewater effluent or ranch run-off, and if so what the mechanisms of endocrine disruption may be. Results/Conclusions Clear links were observed between the four biological scales. At the ranch site, where primarily estrogenic compounds were present, males had significantly higher and females trended towards higher expression of estrogen-responsive genes. At the urban/wastewater outfall site, which is more polluted and contaminated by both estrogens and comparatively higher concentrations of androgens, both males and females had relatively low expression of estrogen-responsive genes, and males had lower gonadal somatic index and higher incidence of severe testicular necrosis. Notably, the proportion of females caught from the urban/wastewater outfall site throughout the spawning season over a two year period was significantly lower than the ranch site and in comparison to historical observations of Menidia sex ratios in other populations. Menidia appear to be highly sensitive to EDCs and our results suggest that the population at the urban/wastewater site is being negatively impacted by these contaminants at several biological scales. Results from this study were also used to parameterize a population model that predicts the conditions under which EDC exposure could contribute to population decline.

Research paper thumbnail of Systems biology meets stress ecology: linking molecular and organismal stress responses in Daphnia magna

Genome Biology, 2008

Background: Ibuprofen and other nonsteroidal anti-inflammatory drugs have been designed to interr... more Background: Ibuprofen and other nonsteroidal anti-inflammatory drugs have been designed to interrupt eicosanoid metabolism in mammals, but little is known of how they affect nontarget organisms. Here we report a systems biology study that simultaneously describes the transcriptomic and phenotypic stress responses of the model crustacean Daphnia magna after exposure to ibuprofen.

Research paper thumbnail of Gene expression responses of threespine stickleback to salinity: implications for salt-sensitive hypertension

Frontiers in Genetics, 2014

Despite recent success with genome-wide association studies (GWAS), identifying hypertension (HTN... more Despite recent success with genome-wide association studies (GWAS), identifying hypertension (HTN)-susceptibility loci in the general population remains difficult. Here, we present a novel strategy to address this challenge by studying salinity adaptation in the threespine stickleback, a fish species with diverse salt-handling ecotypes. We acclimated native freshwater (FW) and anadromous saltwater (SW) threespine sticklebacks to fresh, brackish, and sea water for 30 days, and applied RNA sequencing to determine the gene expression in fish kidneys. We identified 1844 salt-responsive genes that were differentially expressed between FW sticklebacks acclimated to different salinities and/or between SW and FW sticklebacks acclimated to full-strength sea water. Significant overlap between stickleback salt-responsive genes and human genes implicated in HTN was detected (P < 10 −7 , hypergeometric test), suggesting a possible similarity in genetic mechanisms of salt handling between threespine sticklebacks and humans. The overlapping genes included a newly discovered HTN gene-MAP3K15, whose expression in FW stickleback kidneys decreases with salinity. These also included genes located in the GWAS loci such as AGTRAP-PLOD1 and CYP1A1-ULK3, which contain multiple potentially causative genes contributing to HTN susceptibility that need to be prioritized for study. Taken together, we show that stickleback salt-responsive genes provide valuable information facilitating the identification of human HTN genes. Thus, threespine sticklebacks may be used as a model, complementary to existing animal models, in human HTN research.

Research paper thumbnail of TOXICITY TESTING OF GROUNDWATER QUALITY

Water and Environment Journal, 2005

Groundwater is an important resource in the UK, with 45% of public water suppiies in the Thames W... more Groundwater is an important resource in the UK, with 45% of public water suppiies in the Thames Water region derived from subterranean sources. In urban areas. groundwater has been affected by anihropogenic activlties over a long period of time and from a multitude of sources. At present, groundwater quality is assessed using a range of chemical species to determine the extent of contamination. However, analysing a complex mixture of chemicals is timeconsuming and expensive. whereas the use of an ecotoxicity test provides information on (a) the degree of pollution present in the groundwater and (b) the potential effect of that pollution. MicrotoxrM, €cioxrM and Daphnia magna microtests were used in conjunction with standard chemical protocols to assess the contamination of groundwaters from sites throughout the London Borough of Hounsiow and nearby Heathrow Airport. Because of their precision, range of responses and ease of use, Daphnia magna and MicrotoxrM tests are the bioassays that appear to be most effective for assessing groundwater toxicik However, neither test is ideal because It Is also essential to monitor water hardness. EcloxrM does not appear to be suitable for use in groundwaterquality assessment in this area, because it is adversely affected by high total dissolved solids and electrical conductivity.

Research paper thumbnail of Sublethal toxicity of commercial insecticide formulations and their active ingredients to larval fathead minnow (Pimephales promelas)

Science of The Total Environment, 2010

Research paper thumbnail of From ‘Omics to Otoliths: Responses of an Estuarine Fish to Endocrine Disrupting Compounds across Biological Scales

PLoS ONE, 2013

Endocrine disrupting chemicals (EDCs) cause physiological abnormalities and population decline in... more Endocrine disrupting chemicals (EDCs) cause physiological abnormalities and population decline in fishes. However, few studies have linked environmental EDC exposures with responses at multiple tiers of the biological hierarchy, including population-level effects. To this end, we undertook a four-tiered investigation in the impacted San Francisco Bay estuary with the Mississippi silverside (Menidia audens), a small pelagic fish. This approach demonstrated links between different EDC sources and fish responses at different levels of biological organization. First we determined that water from a study site primarily impacted by ranch run-off had only estrogenic activity in vitro, while water sampled from a site receiving a combination of urban, limited ranch run-off, and treated wastewater effluent had both estrogenic and androgenic activity. Secondly, at the molecular level we found that fish had higher mRNA levels for estrogen-responsive genes at the site where only estrogenic activity was detected but relatively lower expression levels where both estrogenic and androgenic EDCs were detected. Thirdly, at the organism level, males at the site exposed to both estrogens and androgens had significantly lower mean gonadal somatic indices, significantly higher incidence of severe testicular necrosis and altered somatic growth relative to the site where only estrogens were detected. Finally, at the population level, the sex ratio was significantly skewed towards males at the site with measured androgenic and estrogenic activity. Our results suggest that mixtures of androgenic and estrogenic EDCs have antagonistic and potentially additive effects depending on the biological scale being assessed, and that mixtures containing androgens and estrogens may produce unexpected effects. In summary, evaluating EDC response at multiple tiers is necessary to determine the source of disruption (lowest scale, i.e. cell line) and what the ecological impact will be (largest scale, i.e. sex ratio).

Research paper thumbnail of The use of image analysis to estimate population growth rate in Daphnia magna

Journal of Applied Ecology, 2006

Population growth rate (PGR) is central to the theory of population ecology and is crucial for pr... more Population growth rate (PGR) is central to the theory of population ecology and is crucial for projecting population trends in conservation biology, pest management and wildlife harvesting. Furthermore, PGR is increasingly used to assess the effects of stressors. Image analysis that can automatically count and measure photographed individuals offers a potential methodology for estimating PGR. 2. This study evaluated two ways in which the PGR of Daphnia magna , exposed to different stressors, can be estimated using an image analysis system. The first method estimated PGR as the ratio of counts of individuals obtained at two different times, while the second method estimated PGR as the ratio of population sizes at two different times, where size is measured by the sum of the individuals' surface areas, i.e. total population surface area. This method is attractive if surface area is correlated with reproductive value (RV), as it is for D. magna , because of the theoretical result that PGR is the rate at which the population RV increases. 3. The image analysis system proved reliable and reproducible in counting populations of up to 440 individuals in 5 L of water. Image counts correlated well with manual counts but with a systematic underestimate of about 30%. This does not affect accuracy when estimating PGR as the ratio of two counts. Area estimates of PGR correlated well with count estimates, but were systematically higher, possibly reflecting their greater accuracy in the study situation. 4. Analysis of relevant scenarios suggested the correlation between RV and body size will generally be good for organisms in which fecundity correlates with body size. In these circumstances, area estimation of PGR is theoretically better than count estimation. 5. Synthesis and applications . There are both theoretical and practical advantages to area estimation of population growth rate when individuals' reproductive values are consistently well correlated with their surface areas. Because stressors may affect both the number and quality of individuals, area estimation of population growth rate should improve the accuracy of predicting stress impacts at the population level.

Research paper thumbnail of Cytochrome P4501A mRNA and protein induction in striped bass (Morone saxatilis)

Fish Physiology and Biochemistry, 2012

The striped bass (Morone saxatilis) supports a valuable recreational fishery and is among the mos... more The striped bass (Morone saxatilis) supports a valuable recreational fishery and is among the most important piscivorous fish of the San Francisco Estuary. This species has suffered a significant decline in numbers over the past decades, and there is indication that contaminants are important contributors. Polycyclic aromatic hydrocarbons (PAHs) and polyhalogenated aromatic hydrocarbons (PHAHs) including PCBs and dioxins are widespread in the estuary, they typically bioaccumulate through trophic levels, reaching highest levels in top predators and are known to affect the fish health and development. The aim of this study was to

Research paper thumbnail of MULTIVARIATE RELATIONSHIPS BETWEEN GROUNDWATER CHEMISTRY AND TOXICITY IN AN URBAN AQUIFER

Environmental Toxicology and Chemistry, 2003

Research paper thumbnail of An assessment of the toxicity of some pesticides and their metabolites affecting a natural aquatic environment using the Microtox™ system

The conservation and preservation of aquatic ecosystems is of utmost importance due to the high d... more The conservation and preservation of aquatic ecosystems is of utmost importance due to the high diversity and density of species and their complex food network. The evaluation of the potential adverse environmental impact caused by pesticides entering water bodies is an important parameter in aquatic toxicity. The toxicity of the insecticide, Fenitrothion and two of its metabolites, 3-methyl-4 nitrophenol and

Research paper thumbnail of Gene expression responses of threespine stickleback to salinity: implications for salt-sensitive hypertension

Frontiers in Genetics

Despite recent success with genome-wide association studies (GWAS), identifying hypertension (HTN... more Despite recent success with genome-wide association studies (GWAS), identifying hypertension (HTN)-susceptibility loci in the general population remains difficult. Here, we present a novel strategy to address this challenge by studying salinity adaptation in the threespine stickleback, a fish species with diverse salt-handling ecotypes. We acclimated native freshwater (FW) and anadromous saltwater (SW) threespine sticklebacks to fresh, brackish, and sea water for 30 days, and applied RNA sequencing to determine the gene expression in fish kidneys. We identified 1844 salt-responsive genes that were differentially expressed between FW sticklebacks acclimated to different salinities and/or between SW and FW sticklebacks acclimated to full-strength sea water. Significant overlap between stickleback salt-responsive genes and human genes implicated in HTN was detected (P < 10−7, hypergeometric test), suggesting a possible similarity in genetic mechanisms of salt handling between threes...

Research paper thumbnail of Acute and Chronic Invertebrate and Fish Toxicity Testing in the Sacramento-San Joaquin Delta 2008-2009, Final Report, Interagency Ecological Program, Sacramento, CA

Research paper thumbnail of 10-Day survival of Hyalella azteca as a function of water quality parameters

Ecotoxicology and environmental safety, 2015

Estuarine systems are among the most impacted ecosystems due to anthropogenic contaminants; howev... more Estuarine systems are among the most impacted ecosystems due to anthropogenic contaminants; however, they present unique challenges to toxicity testing with regard to varying water quality parameters. The euryhaline amphipod species, Hyalella azteca, is widely used in toxicity testing and well suited for testing estuarine water samples. Nevertheless, the influence of relevant water quality parameters on test endpoints must be quantified in order to efficiently use this species for routine monitoring. Here, we studied the influence of five water quality parameters: electrical conductivity, pH, un-ionized ammonia, dissolved oxygen and temperature, on H. azteca survival in a water column toxicity test. A model was developed to quantify and predict the independent and interacting effects of water quality variables on 10-day survival. The model allows simultaneous assessment of multiple potential predictors recorded during the tests. Data used for modeling came from 1089 tests performed ...

Research paper thumbnail of Effect-based tools for monitoring and predicting the ecotoxicological effects of chemicals in the aquatic environment

Sensors (Basel, Switzerland), 2012

Ecotoxicology faces the challenge of assessing and predicting the effects of an increasing number... more Ecotoxicology faces the challenge of assessing and predicting the effects of an increasing number of chemical stressors on aquatic species and ecosystems. Herein we review currently applied tools in ecological risk assessment, combining information on exposure with expected biological effects or environmental water quality standards; currently applied effect-based tools are presented based on whether exposure occurs in a controlled laboratory environment or in the field. With increasing ecological relevance the reproducibility, specificity and thus suitability for standardisation of methods tends to diminish. We discuss the use of biomarkers in ecotoxicology including ecotoxicogenomics-based endpoints, which are becoming increasingly important for the detection of sublethal effects. Carefully selected sets of biomarkers allow an assessment of exposure to and effects of toxic chemicals, as well as the health status of organisms and, when combined with chemical analysis, identificatio...

Research paper thumbnail of Expression of target and reference genes in Daphnia magna exposed to ibuprofen

BMC genomics, 2006

Transcriptomic techniques are now being applied in ecotoxicology and toxicology to measure the im... more Transcriptomic techniques are now being applied in ecotoxicology and toxicology to measure the impact of stressors and develop understanding of mechanisms of toxicity. Microarray technology in particular offers the potential to measure thousands of gene responses simultaneously. However, it is important that microarrays responses should be validated, at least initially, using real-time quantitative polymerase chain reaction (QPCR). The accurate measurement of target gene expression requires normalisation to an invariant internal control e.g., total RNA or reference genes. Reference genes are preferable, as they control for variation inherent in the cDNA synthesis and PCR. However, reference gene expression can vary between tissues and experimental conditions, which makes it crucial to validate them prior to application. We evaluated 10 candidate reference genes for QPCR in Daphnia magna following a 24 h exposure to the non-steroidal anti-inflammatory drug (NSAID) ibuprofen (IB) at 0...

Research paper thumbnail of Chronic toxicity of ibuprofen to Daphnia magna: Effects on life history traits and population dynamics

Toxicology letters, 2007

The non-steroidal anti-inflammatory drug (NSAID) ibuprofen (IB) is a widely used pharmaceutical t... more The non-steroidal anti-inflammatory drug (NSAID) ibuprofen (IB) is a widely used pharmaceutical that can be found in several freshwater ecosystems. Acute toxicity studies with Daphnia magna suggest that the 48h EC(50) (immobilisation) is 10-100 mgIBl(-1). However, there are currently no chronic IB toxicity data on arthropod populations, and the aquatic life impacts of such analgesic drugs are still undefined. We performed a 14-day exposure of D. magna to IB as a model compound (concentration range: 0, 20, 40 and 80 mgIBl(-1)) measuring chronic effects on life history traits and population performance. Population growth rate was significantly reduced at all IB concentrations, although survival was only affected at 80 mgIBl(-1). Reproduction, however, was affected at lower concentrations of IB (14-day EC(50) of 13.4 mgIBl(-1)), and was completely inhibited at the highest test concentration. The results from this study indicate that the long-term crustacean population consequences of a...

Research paper thumbnail of Is there a relationship between soil and groundwater toxicity?

Environmental geochemistry and health, 2003

Part IIA of the Environmental Protection Act 1990 requires environmental regulators to assess the... more Part IIA of the Environmental Protection Act 1990 requires environmental regulators to assess the risk of contaminants leaching from soils into groundwater (DETR, 1999). This newly introduced legislation assumes a link between soil and groundwater chemistry, in which rainwater leaches contaminants from soil into the saturated zone. As the toxicity of both groundwater and overlying soils is dependent upon the chemicals present, their partitioning and their bioavailability, similar patterns of soil, leachates and groundwater toxicity should be observed at contaminated sites. Soil and groundwater samples were collected from different contaminated land sites in an urban area, and used to determine relationships between soil chemistry and toxicity, mobility of contaminants, and groundwater chemistry and toxicity. Soils were leached using water to mimic rainfall, and both the soils and leachates tested using bioassays. Soil bioassays were carried out using Eisenia fetida, whilst groundwat...

Research paper thumbnail of Durieux et al CYP1A 2012 v def

The striped bass (Morone saxatilis) supports a valuable recreational fishery and is among the mos... more The striped bass (Morone saxatilis) supports a valuable recreational fishery and is among the most important piscivorous fish of the San Francisco Estuary. This species has suffered a significant decline in numbers over the past decades, and there is indication that contaminants are important contributors. Polycyclic aromatic hydrocarbons (PAHs) and polyhalogenated aromatic hydrocarbons (PHAHs) including PCBs and dioxins are widespread in the estuary, they typically bioaccumulate through trophic levels, reaching highest levels in top predators and are known to affect the fish health and development. The aim of this study was to

Research paper thumbnail of Haem peroxidase activity in Daphnia magna: a biomarker for sub-lethal toxicity assessments of kerosene-contaminated groundwater

Ecotoxicology (London, England), 2003

A novel biomarker was developed in Daphnia magna to detect organic pollution in groundwater. The ... more A novel biomarker was developed in Daphnia magna to detect organic pollution in groundwater. The haem peroxidase assay, which is an indirect means of measuring oxidase activity, was particularly sensitive to kerosene contamination. Exposure to sub-lethal concentrations of kerosene-contaminated groundwater resulted in a haem peroxidase activity increase by dose with a two-fold activity peak at 25%. Reproduction in D. magna remained unimpaired when exposed to concentrations below 25% for 21 days, and a decline in fecundity was only observed at concentrations above the peak in enzyme activity. The measurement of haem peroxidase activity in D. magna detected sublethal effects of kerosene in just 24 h, whilst offering information on the health status of the organisms. The biomarker may be useful in determining concentrations above which detrimental effects would occur from long-term exposure for fuel hydrocarbons. Moreover, this novel assay detects exposure to chemicals in samples that w...

Research paper thumbnail of Expression and function of ryanodine receptor related pathways in PCB tolerant Atlantic killifish (Fundulus heteroclitus) from New Bedford Harbor, MA, USA

Aquatic Toxicology, 2015

Atlantic killifish (Fundulus heteroclitus) thrive in New Bedford Harbor (NBH), MA, highly contami... more Atlantic killifish (Fundulus heteroclitus) thrive in New Bedford Harbor (NBH), MA, highly contaminated with polychlorinated biphenyls (PCBs). Resident killifish have evolved tolerance to dioxin-like (DL) PCBs, whose toxic effects through the aryl hydrocarbon receptor (AhR) are well studied. In NBH, non-dioxin like PCBs (NDL PCBs), which lack activity toward the AhR, vastly exceed levels of DL congeners yet how killifish counter NDL toxic effects has not been explored. In mammals and fish, NDL PCBs are potent activators of ryanodine receptors (RyR), Ca 2+ release channels necessary for a vast array of physiological processes. In the current study we compared the expression and function of RyR related pathways in NBH killifish with killifish from the reference site at Scorton Creek (SC, MA). Relative to the SC fish, adults from NBH displayed increased levels of skeletal muscle RyR1 protein, and increased levels of FK506-binding protein 12 kDa (FKBP12) an accessory protein essential for NDL PCB-triggered changes in RyR channel function. In accordance with increased RyR1 levels, NBH killifish displayed increased maximal ligand binding, increased maximal response to Ca 2+ activation and increased maximal response to activation by the NDL PCB congener PCB 95. Compared to SC, NBH embryos and larvae had increased levels of mtor and ryr2 transcripts at multiple stages of development, and generations, while levels of serca2 were decreased at 9 days post-fertilization in the F1 and F2 generations. These findings suggest that there are compensatory and heritable changes in RyR mediated Ca 2+ signaling proteins or potential signaling partners in NBH killifish.

Research paper thumbnail of From'omics to otoliths: Using responses to endocrine disrupting compounds at multiple biological scales to predict population dynamics

ABSTRACT Background/Question/Methods Endocrine disrupting chemicals (EDCs) interfere with endogen... more ABSTRACT Background/Question/Methods Endocrine disrupting chemicals (EDCs) interfere with endogenous hormones and cause physiological abnormalities in fishes. Both theory and empirical data confirm that EDCs can also cause declines in fish populations. However, few studies have attempted to link complex environmental EDC mixtures with responses at multiple tiers of the biological hierarchy, including population-level effects. Additionally, the use of environmentally relevant fishes as model species from regions with demonstrated fish population decline is lacking. To this end, we undertook a four-tiered investigation into estrogenic and androgenic EDC effects in Menidia audens (Mississippi silverside), an atherinid distributed throughout the impacted San Francisco Bay (SFB) estuary. Our main objective was to integrate observations at each biological scale with the goal of determining whether the reproductive health of Menidia populations is being negatively impacted by sites receiving either urban run-off and treated wastewater effluent or ranch run-off, and if so what the mechanisms of endocrine disruption may be. Results/Conclusions Clear links were observed between the four biological scales. At the ranch site, where primarily estrogenic compounds were present, males had significantly higher and females trended towards higher expression of estrogen-responsive genes. At the urban/wastewater outfall site, which is more polluted and contaminated by both estrogens and comparatively higher concentrations of androgens, both males and females had relatively low expression of estrogen-responsive genes, and males had lower gonadal somatic index and higher incidence of severe testicular necrosis. Notably, the proportion of females caught from the urban/wastewater outfall site throughout the spawning season over a two year period was significantly lower than the ranch site and in comparison to historical observations of Menidia sex ratios in other populations. Menidia appear to be highly sensitive to EDCs and our results suggest that the population at the urban/wastewater site is being negatively impacted by these contaminants at several biological scales. Results from this study were also used to parameterize a population model that predicts the conditions under which EDC exposure could contribute to population decline.

Research paper thumbnail of Systems biology meets stress ecology: linking molecular and organismal stress responses in Daphnia magna

Genome Biology, 2008

Background: Ibuprofen and other nonsteroidal anti-inflammatory drugs have been designed to interr... more Background: Ibuprofen and other nonsteroidal anti-inflammatory drugs have been designed to interrupt eicosanoid metabolism in mammals, but little is known of how they affect nontarget organisms. Here we report a systems biology study that simultaneously describes the transcriptomic and phenotypic stress responses of the model crustacean Daphnia magna after exposure to ibuprofen.

Research paper thumbnail of Gene expression responses of threespine stickleback to salinity: implications for salt-sensitive hypertension

Frontiers in Genetics, 2014

Despite recent success with genome-wide association studies (GWAS), identifying hypertension (HTN... more Despite recent success with genome-wide association studies (GWAS), identifying hypertension (HTN)-susceptibility loci in the general population remains difficult. Here, we present a novel strategy to address this challenge by studying salinity adaptation in the threespine stickleback, a fish species with diverse salt-handling ecotypes. We acclimated native freshwater (FW) and anadromous saltwater (SW) threespine sticklebacks to fresh, brackish, and sea water for 30 days, and applied RNA sequencing to determine the gene expression in fish kidneys. We identified 1844 salt-responsive genes that were differentially expressed between FW sticklebacks acclimated to different salinities and/or between SW and FW sticklebacks acclimated to full-strength sea water. Significant overlap between stickleback salt-responsive genes and human genes implicated in HTN was detected (P < 10 −7 , hypergeometric test), suggesting a possible similarity in genetic mechanisms of salt handling between threespine sticklebacks and humans. The overlapping genes included a newly discovered HTN gene-MAP3K15, whose expression in FW stickleback kidneys decreases with salinity. These also included genes located in the GWAS loci such as AGTRAP-PLOD1 and CYP1A1-ULK3, which contain multiple potentially causative genes contributing to HTN susceptibility that need to be prioritized for study. Taken together, we show that stickleback salt-responsive genes provide valuable information facilitating the identification of human HTN genes. Thus, threespine sticklebacks may be used as a model, complementary to existing animal models, in human HTN research.

Research paper thumbnail of TOXICITY TESTING OF GROUNDWATER QUALITY

Water and Environment Journal, 2005

Groundwater is an important resource in the UK, with 45% of public water suppiies in the Thames W... more Groundwater is an important resource in the UK, with 45% of public water suppiies in the Thames Water region derived from subterranean sources. In urban areas. groundwater has been affected by anihropogenic activlties over a long period of time and from a multitude of sources. At present, groundwater quality is assessed using a range of chemical species to determine the extent of contamination. However, analysing a complex mixture of chemicals is timeconsuming and expensive. whereas the use of an ecotoxicity test provides information on (a) the degree of pollution present in the groundwater and (b) the potential effect of that pollution. MicrotoxrM, €cioxrM and Daphnia magna microtests were used in conjunction with standard chemical protocols to assess the contamination of groundwaters from sites throughout the London Borough of Hounsiow and nearby Heathrow Airport. Because of their precision, range of responses and ease of use, Daphnia magna and MicrotoxrM tests are the bioassays that appear to be most effective for assessing groundwater toxicik However, neither test is ideal because It Is also essential to monitor water hardness. EcloxrM does not appear to be suitable for use in groundwaterquality assessment in this area, because it is adversely affected by high total dissolved solids and electrical conductivity.

Research paper thumbnail of Sublethal toxicity of commercial insecticide formulations and their active ingredients to larval fathead minnow (Pimephales promelas)

Science of The Total Environment, 2010

Research paper thumbnail of From ‘Omics to Otoliths: Responses of an Estuarine Fish to Endocrine Disrupting Compounds across Biological Scales

PLoS ONE, 2013

Endocrine disrupting chemicals (EDCs) cause physiological abnormalities and population decline in... more Endocrine disrupting chemicals (EDCs) cause physiological abnormalities and population decline in fishes. However, few studies have linked environmental EDC exposures with responses at multiple tiers of the biological hierarchy, including population-level effects. To this end, we undertook a four-tiered investigation in the impacted San Francisco Bay estuary with the Mississippi silverside (Menidia audens), a small pelagic fish. This approach demonstrated links between different EDC sources and fish responses at different levels of biological organization. First we determined that water from a study site primarily impacted by ranch run-off had only estrogenic activity in vitro, while water sampled from a site receiving a combination of urban, limited ranch run-off, and treated wastewater effluent had both estrogenic and androgenic activity. Secondly, at the molecular level we found that fish had higher mRNA levels for estrogen-responsive genes at the site where only estrogenic activity was detected but relatively lower expression levels where both estrogenic and androgenic EDCs were detected. Thirdly, at the organism level, males at the site exposed to both estrogens and androgens had significantly lower mean gonadal somatic indices, significantly higher incidence of severe testicular necrosis and altered somatic growth relative to the site where only estrogens were detected. Finally, at the population level, the sex ratio was significantly skewed towards males at the site with measured androgenic and estrogenic activity. Our results suggest that mixtures of androgenic and estrogenic EDCs have antagonistic and potentially additive effects depending on the biological scale being assessed, and that mixtures containing androgens and estrogens may produce unexpected effects. In summary, evaluating EDC response at multiple tiers is necessary to determine the source of disruption (lowest scale, i.e. cell line) and what the ecological impact will be (largest scale, i.e. sex ratio).

Research paper thumbnail of The use of image analysis to estimate population growth rate in Daphnia magna

Journal of Applied Ecology, 2006

Population growth rate (PGR) is central to the theory of population ecology and is crucial for pr... more Population growth rate (PGR) is central to the theory of population ecology and is crucial for projecting population trends in conservation biology, pest management and wildlife harvesting. Furthermore, PGR is increasingly used to assess the effects of stressors. Image analysis that can automatically count and measure photographed individuals offers a potential methodology for estimating PGR. 2. This study evaluated two ways in which the PGR of Daphnia magna , exposed to different stressors, can be estimated using an image analysis system. The first method estimated PGR as the ratio of counts of individuals obtained at two different times, while the second method estimated PGR as the ratio of population sizes at two different times, where size is measured by the sum of the individuals' surface areas, i.e. total population surface area. This method is attractive if surface area is correlated with reproductive value (RV), as it is for D. magna , because of the theoretical result that PGR is the rate at which the population RV increases. 3. The image analysis system proved reliable and reproducible in counting populations of up to 440 individuals in 5 L of water. Image counts correlated well with manual counts but with a systematic underestimate of about 30%. This does not affect accuracy when estimating PGR as the ratio of two counts. Area estimates of PGR correlated well with count estimates, but were systematically higher, possibly reflecting their greater accuracy in the study situation. 4. Analysis of relevant scenarios suggested the correlation between RV and body size will generally be good for organisms in which fecundity correlates with body size. In these circumstances, area estimation of PGR is theoretically better than count estimation. 5. Synthesis and applications . There are both theoretical and practical advantages to area estimation of population growth rate when individuals' reproductive values are consistently well correlated with their surface areas. Because stressors may affect both the number and quality of individuals, area estimation of population growth rate should improve the accuracy of predicting stress impacts at the population level.

Research paper thumbnail of Cytochrome P4501A mRNA and protein induction in striped bass (Morone saxatilis)

Fish Physiology and Biochemistry, 2012

The striped bass (Morone saxatilis) supports a valuable recreational fishery and is among the mos... more The striped bass (Morone saxatilis) supports a valuable recreational fishery and is among the most important piscivorous fish of the San Francisco Estuary. This species has suffered a significant decline in numbers over the past decades, and there is indication that contaminants are important contributors. Polycyclic aromatic hydrocarbons (PAHs) and polyhalogenated aromatic hydrocarbons (PHAHs) including PCBs and dioxins are widespread in the estuary, they typically bioaccumulate through trophic levels, reaching highest levels in top predators and are known to affect the fish health and development. The aim of this study was to

Research paper thumbnail of MULTIVARIATE RELATIONSHIPS BETWEEN GROUNDWATER CHEMISTRY AND TOXICITY IN AN URBAN AQUIFER

Environmental Toxicology and Chemistry, 2003