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Genetically Engineered Erwinia carotovora: Survival, Intraspecific Competition, and Effects upon Selected Bacterial Genera.

Environmental use of genetically engineered microorganisms has raised concerns about potential ecological impact. This research evaluated the survival, competitiveness, and effects upon selected bacterial genera of wild-type and genetically engineered Erwinia carotovora subsp. carotovora to ascertain if differences between the wild-type and genetically engineered strains exist in soil microcosms. The engineered strain contained a chromosomally inserted gene for kanamycin resistance. No significant differences in survival in nonsterile soil over 2 months or in the competitiveness of either strain were observed when the strains were added concurrently to microcosms. For reasons that remain unclear, the engineered strain did survive longer in sterilized soil. The effects of both strains on total bacteria, Pseudomonas and Staphylococcus strains, and actinomycetes were observed. While some apparent differences were observed, they were not statistically significant. A better understanding of the microbial ecology of engineered bacteria, especially pathogens genetically altered for use as biological control agents, is essential before commercial applications can be accomplished.

Journal Article↗

Conflict monitoring versus selection-for-action in anterior cingulate cortex.

The anterior cingulate cortex (ACC), on the medial surface of the frontal lobes of the brain, is widely believed to be involved in the regulation of attention. Beyond this, however, its specific contribution to cognition remains uncertain. One influential theory has interpreted activation within the ACC as reflecting 'selection-for-action', a set of processes that guide the selection of environmental objects as triggers of or targets for action. We have proposed an alternative hypothesis, in which the ACC serves not to exert top-down attentional control but instead to detect and signal the occurrence of conflicts in information processing. Here, to test this theory against the selection-for-action theory, we used functional magnetic resonance imaging to measure brain activation during performance of a task where, for a particular subset of trials, the strength of selection-for-action is inversely related to the degree of response conflict. Activity within the ACC was greater during trials featuring high levels of conflict (and weak selection-for-action) than during trials with low levels of conflict (and strong selection-for-action), providing evidence in favour of the conflict-monitoring account of ACC function.

Adult↗

Sperm-limited fecundity in nematodes: how many sperm are enough?

The Bateman principle, which holds that oocytes are the limiting gamete in reproduction, is violated in a variety of species. Self-fertilizing hermaphrodites of the nematode Caenorhabditis elegans provide an example of a system in which sperm number limits lifetime reproductive output, in this species due to the protandrous nature of sperm production that in turn delays the onset of fertilization. This reproductive delay forms the basis of a trade-off between generation time and total fecundity, in which sperm number plays a pivotal role. I use an age-structured population model to describe the number of sperm that maximize fitness, given larval development time and rates of gamete production. The model predicts the evolution of sperm numbers that are consistent with empirical data for C. elegans provided that precocious larval sperm production is taken into account. Several testable hypotheses follow from the model regarding how natural selection and environmental variation may influence patterns of sperm production among populations or species with a similar mode of reproduction.

Age Factors↗

Molecular and immunochemical characteristics of monoclonal and recombinant antibodies selective for the triazine herbicide simetryn and application to environmental analysis.

A monoclonal antibody (mab) selective for the thiomethyl-s-triazine herbicide simetryn was obtained and characterized in enzyme-linked immunosorbent assay (ELISA). An IC(50) value for simetryn was 8.5 ng/mL, and the detection range extended from 1.1 to 70 ng/mL in ELISA. The cDNAs encoding variable heavy chain (VH) and variable light chain (VL) of the mab were cloned to produce various recombinant antibodies. Single-chain variable fragment (scFv) antibodies derived from the mab were characterized in ELISA and showed similar reactivities and specificities to the parent mab. A urea denaturation test revealed that the scFv antibodies bound to simetryn were more stable than those in the absence of antigen. A sandwich ELISA based on VH and VL fragments of the mab was successfully developed and showed similar sensitivity to those based on the mab and scFv antibodies in ELISA. In the recovery experiments using spiked environmental samples, the results obtained in ELISA based on the mab were favorably correlated with those by HPLC.

Amino Acid Sequence↗

Technical report: precautions regarding the use of aerosolized antibiotics. Committee on Infectious Diseases and Committee on Drugs.

In 1998, the Food and Drug Administration (FDA) approved the licensure of tobramycin solution for inhalation (TOBI). Although a number of additional antibiotics, including other aminoglycosides, beta-lactams, antibiotics in the polymyxin class, and vancomycin, have been administered as aerosols for many years, none are approved by the FDA for administration by inhalation. TOBI was approved by the FDA for the maintenance therapy of patients 6 years or older with cystic fibrosis (CF) who have between 25% and 75% of predicted forced expiratory volume in 1 second (FEV(1)), are colonized with Pseudomonas aeruginosa, and are able to comply with the prescribed medical regimen. TOBI was not approved for the therapy of acute pulmonary exacerbations in patients with CF nor was it approved for use in patients without CF. Currently, no other antibiotics are approved for administration by inhalation to patients with or without CF. The purpose of this statement is to briefly summarize the data that supported approval for licensure of TOBI and to provide recommendations for its safe use. The pharmacokinetics of inhaled aminoglycosides and problems associated with aerosolized antibiotic treatment, including environmental contamination, selection of resistant microbes, and airway exposure to excipients in intravenous formulations, will be discussed.

Administration, Inhalation↗

An overview of the BEEP Stavanger workshop.

Within the three-year European Research Project BEEP (Biological Effects of Environmental Pollution in marine ecosystems), a workshop was carried out at the facility of Akvamiljø a.s. with the scientific support of IRIS (International Research Institute of Stavanger) researchers. Validation of newly developed biomarkers, especially at the molecular level, was the core part of the activity. Two large scale mesocosm exposures were done and samples shared between the participants (about 30 Institute), allowing an integrated approach for studying the effects of several pollutants selected as environmentally important issues (i.e. PAHs, alkyl phenols, bisphenol A, diallyl phthalate and polybrominated diphenyl ether). In addition, a field study (Visne, Norway) has been performed for the validations of biomarkers under environmental conditions. The Stavanger workshop demonstrates the importance of collaboration between researchers to establish common ways to proceed in biomarker analysis. The common exposures encourage a joint strategy for planning biomonitoring activity. A clarification in this direction is needed to coordinate results from numerous studies about the environmental impact of pollutants which are currently carried out all over Europe and around the world. Expertise in various disciplines is required to plan and perform a successful monitoring activity and to study/predict the effect of environmental pollutants. Biologists, chemists, statisticians and environmental researchers should always be included. The Stavanger workshop also indicated the importance of developing multivariate statistical methods to enable interpretation of complex data sets. It is of interest to develop statistical tools which can integrate the results from a battery of biomarkers in order to obtain a more complete picture of environmental impact.

Animals↗

Long-term mortality study of steelworkers. IX. Mortality patterns among sheet and tin mill workers.

As a result of findings of an earlier report in this series, this study examines the updated cause-specific mortality of men employed in the sheet and tin mill areas of the steel industry. In order to investigate possible relationships between occupational responsibilities or exposures and mortality from specific causes, the sheet and tin mills have been subdivided into 13 mutually exclusive work areas. Detailed analysis is limited primarily to white workers due to the small number of nonwhites in these areas. The most important observations are: 1. Increased overall mortality appears for men employed in 1953 in the sheet finishing and shipping area, confirming the findings of Lloyd, et al. The earlier observation of a significant excess in deaths from vascular lesions of the central nervous system does not hold over time. The previously noted excess for this cause may be related to selective factors or an extreme chance observation. The excess in mortality from all causes of death, which occurs over several disease categories, may not be a result of occupational exposures, but rather some selectivity. 2. Significant excesses in mortality from arteriosclerotic heart disease are noted among men employed in batch pickling and sheet dryer operations, which is in agreement with the earlier findings. Increased risks of dying from hypertensive heart disease are seen in the coating area. 3. Cancer of the lymphatic and hematopoietic tissues is found to be a significant source of excess mortality for workers in the heat treating and forging and tin finishing and shipping work areas. 4. Steelworkers employed in the annealing-normalizing work area show an excess in deaths from nonmalignant respiratory diseases, primarily pneumonia. Further study in these areas should attempt to investigate whether factors in the work environment may be responsible for the observed excess mortalities. More specifically, work should be done to find out whether men employed in heat treating and forging and tin finishing and shipping work in close proximity to chemicals or radiation exposure and whether workers employed in the annealing-normalizing area are exposed to any kind of oil, vapor, or chemical which might be irritating or infectious to the respiratory system. A similar analysis for men working in the batch pickling and sheet dryers and coating areas would also be worthwhile. The main emphasis of any future study should lie upon investigating whether the observed excess mortalities are due to any environmental factor, selection for health, or random fluctuation.

Alloys↗

Multivariate Effects of SNPs on Environmental Streptococcal Mastitis Evaluated With an NGS-Based Association Study Using Targeted Resequencing in the Bovine MHC Region.

Mastitis is an inflammatory reaction caused by bacterial infection of the teat, and a relationship between its onset and cattle major histocompatibility complex (BoLA) region has been reported. However, no comprehensive genetic analysis of mastitis caused by environmental streptococci has been reported. Here, we resequenced the BoLA region using a hybridisation capture target next-generation sequencing (NGS) method to identify disease susceptibility markers mapped to the BoLA region in environmental streptococcal mastitis. This study examined 75 cows with mastitis caused by environmental streptococci selected from 1641 cows with mastitis and 222 healthy cows without mastitis in Japan. Targeted sequences obtained from MiSeq NGS were aligned to the bovine reference genome (ARS-UCD1.2/bosTau9), and 2,920,355 variants were detected within the BoLA region of the 297 Holstein cattle. In an association study using 2264 variants after quality control, the top 20 variants with the lowest P values were selected and assigned to the 18 surrounding candidate genes, and a gene network analysis of these genes resulted in the narrowing down of five candidate genes POU5F1, IER3, GNL1, ABCF1, and PRR3. Multivariate effect analysis of all 6 SNPs associated with these 5 genes revealed that they were significantly correlated with mastitis, indicating that they were useful for classification of mastitis-resistant and mastitis-susceptible cattle. This is the first report to identify SNPs associated with environmental streptococcal mastitis with an NGS-based association study using targeted resequencing in the BoLA region, and understanding host factors may provide important clues for mastitis control.

Animals↗

Spouse concordance of plasma cholesterol and triglyceride.

Spouse concordance for plasma cholesterol and triglyceride was studied in 1971 spouse pairs including 545 spouse pairs married 40 or more years. Both lipids showed modest but statistically significant age corrected Kendall Tau-B correlations; 0.046 (P = 0.005) for cholesterol and 0.070 (P = 0.0001) for triglycerides. Concordance was not explained by season of sampling or by selective mortality. Relative weight was positively correlated in spouse pairs and appeared to explain much of the observed correlations in cholesterol and triglycerides. Spouse pairs also showed positive correlations for cigarette smoking and egg and milk consumption, though these behavioral variables explained little of the concordance for cholesterol and triglycerides. Lipid concordance was highest in marriages of the shortest (1-9 yr) and longest (40 + yr) durations, as was relative weight concordance, suggesting an interaction between duration of marriage and environmental and/or selective factors affecting plasma lipids.

Animals↗

A dual dominant selection, dual marker gene amplification model for environmental surveillance of recombinant viral vaccines.

A simple dual dominant selection marker gene model is proposed for recombinant DNA (rDNA) viral vaccines. Marker genes are also used as targets for in vitro DNA amplification (polymerase chain reaction). Environmentally stable and destabilized rDNA viruses are easily recovered and identified. Use of the model would facilitate evaluation of pressing environmental and safety issues surrounding release of rDNA herpesviruses and poxviruses into the biosphere.

DNA, Recombinant↗

Hemolytic-Uremic Syndrome and Escherichia coli O121 at a Lake in Connecticut, 1999.

OBJECTIVE: Non-O157 Shiga toxin-producing Escherichia coli (STEC) have emerged as an important public health problem. Outbreaks attributed to non-O157 STEC rarely are reported. In 1999, follow-up of routine surveillance reports of children with hemolytic- uremic syndrome (HUS) identified a small cluster of 3 cases of HUS, all of whom had spent overlapping time in a Connecticut lake community in the week before onset of symptoms. We conducted an investigation to determine the magnitude and source of the outbreak and to determine risk factors associated with the transmission of illness. METHODS: We conducted a cohort study and an environmental investigation. The study population included all people who were at the lake in a defined geographic area during July 16-25, 1999. This time and area were chosen on the basis of interviews with the 3 HUS case-patients. A case was defined as diarrhea (>/=3 loose stools/d for >/=3 days) in a person who was at the lake during July 16-25, 1999. Stool samples were requested from any lake resident with diarrheal illness. Stools were cultured for Salmonella, Shigella, Campylobacter, and E coli O157. Broth cultures of stools were tested for Shiga toxin. Case-patients were asked to submit a serum specimen for antibody testing to lipopolysaccharides of selected STEC. Environmental samples from sediment, drinking water, lake water, and ice were obtained and cultured for E coli and tested for Shiga toxin. An environmental evaluation of the lake was conducted to identify any septic, water supply system, or other environmental condition that could be related to the outbreak. RESULTS: Information was obtained for 436 people from 165 (78%) households. Eleven (2.5%) people had illnesses that met the case definition, including the 3 children with HUS. The attack rate was highest among those who were younger than 10 years and who swam in the lake on July 17 or 18 (12%; relative risk [RR]: 7.3). Illness was associated with swimming (RR = 8.3) and with swallowing water while swimming (RR = 7.0) on these days. No person who swam only after July 18 developed illness. Clinical characteristics of case-patients included fever (27%), bloody diarrhea (27%), and severe abdominal cramping (73%). Only the 3 children with HUS required hospitalization. No bacterial pathogen was isolated from the stool of any case-patient. Among lake residents outside the study area, E coli O121:H19 was obtained from a Shiga toxin-producing isolate from a toddler who swam in the lake. Serum was obtained from 7 of 11 case-patients. Six of 7 case-patients had E coli O121 antibody titers that ranged from 1:320 to >1:20 480. E coli indicative of fecal contamination was identified from sediment and water samples taken from a storm drain that emptied into the beach area and from a stream bed located between 2 houses, but no Shiga toxin-producing strain was identified. CONCLUSIONS: Our findings are consistent with a transient local beach contamination in mid-July, probably with E coli O121:H19, which seems to be able to cause severe illness. Without HUS surveillance, this outbreak may have gone undetected by public health officials. This outbreak might have been detected sooner if Shiga toxin screening had been conducted routinely in HUS cases. Laboratory testing that relies solely on the inability of an isolate to ferment sorbitol will miss non-O157 STEC, such as E coli O121. Serologic testing can be used as an adjunct in the diagnosis of STEC infections. Lake-specific recommendations included education, frequent water sampling, and alternative means for toddlers to use lake facilities.

Adolescent↗

Intron length and accelerated 3' gene evolution.

Genetic evolution depends in part upon a balance between negative selection and environmentally driven mutation. To explore whether this balance is affected by gene structure, we have used phylogenetic data mining to compare gene compositions across a range of species. Here we show that genomes of higher species exhibit a greater frequency of 5' CpG islands and of CpG-->TpG/CpA transitions. This latter mutational pattern exhibits a 5'-to-3' trend in higher species, consistent with a length-dependent effect on methylation-dependent CpG suppression. Associated strand asymmetry (TpG>CpA) declines with gene length, implying attenuation of transcription-coupled repair 3' to introns. A sharp 3' rise in coding region single-nucleotide polymorphism frequency further supports a mechanistic role for intron length in promoting genetic variation by reducing repair and/or weakening negative selection. Consistent with this, the Ka/Ks ratio of 3' exons exceeds that of centrally located exons in intron-containing, but not in intronless, genes (p<0.0003). We conclude that the efficiency of transcription-coupled repair decreases with gene length, suggesting in turn that 3' gene evolution is accelerated both by introns and by gene methylation.

3' Untranslated Regions↗

Lipid remodeling in wild and selectively bred hard clams at low temperatures in relation to genetic and physiological parameters.

A temperature decrease usually induces an ordering effect in membrane phospholipids, which can lead to membrane dysfunction. Poikilotherms inhabiting eurythermal environments typically counteract this temperature effect by remodeling membrane lipids as stipulated in the homeoviscous adaptation theory (HVA). Hard clams, Mercenaria mercenaria, can suffer high overwintering mortalities in the Gulf of St Lawrence, Canada. The selectively bred M. mercenaria var. notata can have higher overwintering mortalities than the wild species, thus suggesting that the two varieties have different degrees of adaptation to low temperatures. The objective of this study was to investigate the changes in lipid composition of soft tissues in wild and selected hard clams in relation to their metabolic and genetic characteristics. Clams were placed at the northern limit of their distribution from August 2003 to May 2004; they were exposed to a gradual temperature decrease and then maintained at <0 degrees C for 3.5 months. This study is the first to report a major remodeling of lipids in this species as predicted by HVA; this remodeling involved a sequential response of the phospholipid to sterol ratio as well as in levels of 22:6n-3 and non-methylene interrupted dienoic fatty acids. Hard clams showed an increase in 20:5n-3 as temperature decreased, but this was not maintained during overwintering, which suggests that 20:5n-3 may have been used for eicosanoid biosynthesis as a stress response to environmental conditions. Selectively bred hard clams were characterized by a higher metabolic demand and a deviation from Hardy-Weinberg equilibrium at several genetic loci due to a deficit in heterozygote frequency compared with wild clams, which is believed to impose additional stress and render these animals more vulnerable to overwintering mortality. Finally, an intriguing finding is that the lower metabolic requirements of wild animals coincide with a lower unsaturation index of their lipids, as predicted by Hulbert's theory of membranes as pacemakers of metabolism.

Animals↗

Assessment of anthracene toxicity toward environmental eukaryotic microorganisms: Tetrahymena pyriformis and selected micromycetes.

The toxicity of anthracene, a nonmutagenic, noncarcinogenic, low-molecular-weight polycyclic aromatic hydrocarbon present in the environment, was assessed using a ciliated protozoan, Tetrahymena pyriformis, and a selection of 10 micromycetes from different taxonomic groups living in two environmental compartments, namely aquatic and soil ecosystems. With T. pyriformis, a concentration-dependent inhibitory effect was shown on the cell proliferation rate, IC(50) = 33.40+/-4.84 mg/L (kinetic method). Phagocytosis of nonsoluble anthracene was confirmed by the presence of digestive vacuoles with photon microscopy. In fungi, anthracene did not exhibit a fungicide effect but showed a fungistatic action. Except for Absidia fusca and Cladosporium herbarum, the micromycetes showed a concentration-dependent decrease in growth. From IC(50) values determined by endpoint or kinetic methods, Rhodotorula glutinis and all of the Ascomycotina (except for Penicillium chrysogenum) were the most sensitive species, while Phanerochaete chrysosporium, P. chrysogenum, and the two Deuteromycotina were more resistant to anthracene. Our discussion focuses on the evaluation of toxicity by the two methods used for the calculation of the IC(50) values (endpoint and kinetic), the advantages of studying growth by a kinetic method (the dynamic aspect), and a comparison of toxicity to T. pyriformis and the different micromycetes.

Animals↗

Environmental changes in the North Atlantic Region: SCANNET as a collaborative approach for documenting, understanding and predicting changes.

The lands surrounding the North Atlantic Region (the SCANNET Region) cover a wide range of climate regimes, physical environments and availability of natural resources. Except in the extreme North, they have supported human populations and various cultures since at least the end of the last ice age. However, the region is also important at a wider geographical scale in that it influences the global climate and supports animals that migrate between the Arctic and all the other continents of the world. Climate, environment and land use in the region are changing rapidly and projections suggest that global warming will be amplified there while increasing land use might dramatically reduce the remaining wilderness areas. Because much of the region is sparsely populated--if populated at all--observational records of past environmental changes and their impacts are both few and of short duration. However, it is becoming very important to record the changes that are now in progress, to understand the drivers of these changes, and to predict future consequences of the changes. To facilitate research into understanding impacts of global change on the lands of the North Atlantic Regions, and also to monitor changes in real time, an EU-funded network of research sites and infrastructures was formed in 2000: this was called SCANNET--SCANdinavian/North European NETwork of Terrestrial Field Bases. SCANNET currently consists of 9 core sites and 5 sites within local networks that together cover the broad range of current climate and predicted change in the region. Climate observations are well replicated across the network, whereas each site has tended to select particular environmental and ecological subjects for intensive observation. This provides diversity of both subject coverage and expertise. In this paper, we summarize the findings of SCANNET to-date and outline its information bases in order to increase awareness of data on environmental change in the North Atlantic Region. We also identify important gaps in our understanding and identify where the roles of existing infrastructures and activities represented by SCANNET can facilitate future research, monitoring and ground-truthing activities.

Agriculture↗

Integrating remote sensing approach with pollution monitoring tools for aquatic ecosystem risk assessment and management: a case study of Lake Victoria (Uganda).

Aquatic ecosystems around the world, lake, estuaries and coastal areas are increasingly impacted by anthropogenic pollutants through different sources such as agricultural, industrial and urban discharges, atmospheric deposition and terrestrial drainage. Lake Victoria is the second largest lake in the world and the largest tropical lake. Bordered by Tanzania, Uganda, and Kenya, it provides a livelihood for millions of Africans in the region. However, the lake is under threat from eutrophication, a huge decline in the number of native fish species caused by several factors including loss of biodiversity, over fishing and pollution has been recently documented. Increasing usage of pesticides and insecticides in the adjacent agricultural areas as well as mercury contamination from processing of gold ore on the southern shores are currently considered among the most emergent phenomena of chemical contamination in the lake. By the application of globally consistent and comprehensive geospatial data-sets based on remote sensing integrated with information on heavy metals accumulation and insecticides exposure in native and alien fish populations, the present study aims at assessing the environmental risk associated to the contamination of the Lake Victoria water body on fish health, land cover distribution, biodiversity and the agricultural area surrounding the lake. By the elaboration of Landsat 7 TM data of November 2002 and Landsat 7 TM 1986 we have calculated the agriculture area which borders the Lake Victoria bay, which is an upland plain. The resulting enhanced nutrient loading to the soil is subsequently transported to the lake by rain or as dry fall. The data has been inserted in a Geographical information System (ARCGIS) to be upgraded and consulted. Heavy metals in fish fillets showed concentrations rather low except for mercury being higher than others as already described in previous investigations. In the same tissue, cholinesterases activity (ChE) as an indicator of insecticides exposure showed significant differences among fish species in both activity and sensitivity of selected inhibitor insecticides. This integrated approach aims at identifying and quantifying selected aquatic environmental issues which integrated with monitoring techniques such as contaminant concentrations and biological responses to insecticides exposure in fish populations will provide a scientific basis for aquatic zones management and assist in policy formulations at the national and international levels.

Animals↗

Enantioselective silyl protection of alcohols catalysed by an amino-acid-based small molecule.

Reliable, selective and environmentally friendly chemical transformations are crucial to the development of new therapeutics and the design of novel materials. Chiral catalysts that can be easily prepared and used to obtain organic molecules of high enantiomeric purity are critical to modern chemical synthesis. The development of protecting groups that shield reactive functionalities has also proved indispensable in the preparation of complex biologically active molecules. Here we present a chiral catalyst that promotes the enantioselective protection of a secondary alcohol as one of the most commonly used protected forms of an alcohol: a silyl ether. The catalyst is a small, simple molecule that can be prepared in three steps from commercial materials without the need for rigorously controlled conditions. Enantioselective silylations are performed with commercial silyl chlorides and produce yields of up to 96 per cent at an enantiomeric ratio of up to 98:2. Chiral catalysts for selective formation of commonly used protecting groups such as silyl ethers should significantly enhance the ability of chemical synthesis to deliver, in a more practical and efficient manner, important organic molecules.

Alcohols↗

The relative estrogenic activity of technical toxaphene mixture and two individual congeners.

Toxaphene is the most abundant persistent organic pollutant in the Arctic and in the Great Lakes. Toxaphene technical mixture (Tox) applied as a pesticide consists of over 800 congeners. Through processes of environmental degradation, selected metabolism, and bioaccumulation, two congeners are prominent in humans; 2-exo,3-endo,5-exo,6-endo,8,8,10,10-octachlorobornane (T2 or Parlar 26) and 2-exo,3-endo,5-exo,6-endo,8,8,9,10, 10-nonachlorocamphene (T12 or Parlar 50). The MCF7-E3 human breast cancer cell model was used to screen for the estrogenic activities of Tox, T2, and T12. A concentration of 10 microM was required by all three compounds to elicit an estrogenic response as indicated by a proliferative effect (PE) upon the cells. The congeners, however, showed significantly different PEs from Tox. Both T2 and T12 had a lower PE (16 and 30%) and than Tox, and T2 had a higher PE than T12 (19%). Results from binary combination studies showed that the effects of Tox, T2, and T12 were additive. Tox, T2, and T12 had no significant effects on estrogen receptor and progesterone receptor levels. Our results suggest that the two environmental prevalent congeners had lower estrogenic activities than Tox and there is no synergistic effect.

Camphanes↗