Generating polyketide diversity in Dictyostelium: a Steely hybrid polyketide synthase produces alternate products at different developmental stages.

The soil is a rich ecosystem where many ecological interactions are mediated by small molecules, and in which amoebae are low-level predators and also prey. The social amoeba Dictyostelium discoideum has a high genomic potential for producing polyketides to mediate its ecological interactions, including the unique 'Steely' enzymes, consisting of a fusion between a fatty acid synthase and a chalcone synthase. We report here that D. discoideum further increases its polyketide potential by using the StlB Steely enzyme, and a downstream chlorinating enzyme, to make both a chlorinated signal molecule, DIF-1, during its multi-cellular development, and a set of abundant polyketides in terminally differentiated stalk cells. We identify one of these as a chlorinated dibenzofuran with potent anti-bacterial activity. To do this, StlB switches expression from prespore to stalk cells in late development and is cleaved to release the chalcone synthase domain. Expression of this domain alone in StlB null cells allows synthesis of the stalk-associated, chlorinated polyketides. Thus, by altered expression and processing of StlB, cells make first a signal molecule, and then abundant secondary metabolites, which we speculate help to protect the mature spores from bacterial infection.

Assessing Benthic Bioaccumulation of Polychlorinated Dioxins/Furans and Polychlorinated Biphenyls in the Lower Passaic River (NJ, USA) Based on In Situ Passive Sampling.

Passive sampling has emerged as a promising tool to assess the presence of hydrophobic organic contaminants (HOC) in water, sediment, and biota, such as polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs) or polychlorinated biphenyls (PCBs). Previous work evaluated the ability of passive samplers to predict the bioavailability of sedimentary HOCs mostly in the laboratory, often for marine organisms. The present study assessed the use of low-density polyethylene (LDPE) to derive freely dissolved concentrations of PCDD/Fs and PCBs in porewater in situ versus ex situ and in river water. An LDPE-based multisampler system was deployed at 4 locations along the lower Passaic River (NJ, USA) in sediment and the water column, where sediment and benthic species samples were also collected. Good agreement was generally observed for PCDD/F and PCB concentrations comparing in situ and ex situ approaches (within 0.30-39%). Significant linear relationships were derived between log LDPE-based and log lipid-based concentrations of PCDD/Fs and PCBs. The in situ multisampler system showed promise to derive HOC concentrations in porewater and river water and to predict the bioaccumulation potential of HOCs in benthic biota. Environ Toxicol Chem 2020;39:1174-1185. © 2020 SETAC.

Perinatal polychlorinated biphenyls and polychlorinated dibenzofurans exposure are associated with DNA methylation changes lasting to early adulthood: Findings from Yucheng second generation.

Epigenome-wide DNA methylation has not been studied in men perinatally exposed to PCBs and dioxins. Therefore, we examined whether perinatal exposure to polychlorinated biphenyls (PCBs) and polychlorinated dibenzofurans (PCDFs) induces sustained methylation changes lasting to early adulthood. We used the Illumina HumanMethylation450 BeadChip to assess DNA methylation in whole blood among Yucheng second generation (people perinatal exposed to high PCBs and PCDFs) compared with referents. Thirty male offspring from the Yucheng cohort were randomly selected and matched with 30 male offspring from the Yucheng' neighborhood referents with similar backgrounds. Methylation differences between the Yucheng second generation and non-exposed referents were identified using a P value < 1.06 × 10-7. Differential DNA methylation with epigenome-wide statistical significance was observed for 20 CpGs mapped to 11 genes, and 19 CpGs were correlated with gestational levels of PCBs or PCDF toxic equivalency (PCDF-TEQ) with the same direction of effect. Among the 11 genes, AHRR and CYP1A1 are involved in the aryl hydrocarbon receptor signaling pathway known to mediate dioxin toxicity. MYO1G, FRMD4A, ARL4C, OLFM1, and WWC3 were previously reported to be related to carcinogenesis. This is the first study examining genome-wide DNA methylation among people perinatally exposed to high concentrations of PCBs and PCDFs. We observed novel differential methylation of several genes, indicating that modifications of DNA methylation associated with perinatal PCB and PCDF exposure may persist in exposed offspring for more than 20 years. Furthermore, involvement of several carcinogesis-related genes suggested a potential in utero epigenetic mechanisms.

Levels of regulated POPs in fish samples from the Sava River Basin. Comparison to legislated quality standard values.

Fish samples of different species (i.e. rainbow trout (Onchorhynchus mykiss), barbel (Barbus barbus) and European chub (Squalius cephalus)) were collected from the Sava River Basin for a preliminary investigation of the levels of PCDD/Fs, PCBs, PBDEs and PFAS as a whole. Concentrations of PCDD/Fs, in terms of pg WHO-TEQ/g ww, were below the maximum limit established at the Commission Regulation (EU) No 1259/2011. On the contrary, when DL-PCBs were also included, levels increase up to 11.7 pg WHO-TEQPCDD/Fs+DL-PCBs/g ww in a particular case, with two samples out of a total of ten exceeding the maximum set at this EU Regulation and the EQS established at the European Directive regarding priority substances in the field of water policy (0.0065 ng WHO-TEQPCDD/Fs+DL-PCBs/g ww). A similar trend was also observed for NDL-PCBs, whit the same two samples, from the lower stretch of the river basin, exceeding the maximum limit allowed at the EU Regulation (125 ng/g ww). For PBDEs, levels found in all the samples exceeded the EQS (0.0085 ng/g ww) up to more than a thousand times and 40% of the samples presented PFOS values above the EQS. Data from this study were compared to values reported at the literature for fish from other geographical areas.

Comparative study of persistent organic pollutant (POP) (chlorinated pesticides, PCBs, and dioxins/furans) concentrations in cancer-affected human organs with those of healthy organs.

The concentrations of POPs (persistent organic pollutants) including 16 compounds of OCPs, 12 dioxin-like PCBs congeners, and 17 PCDDs/Fs congeners were determined in 46 human adipose tissue samples gathered from Jordanian citizens. Thirteen adipose tissue samples of healthy people were collected from Jordan University Hospital and 33 adipose tissue samples of cancer-affected patients were collected from King Hussein Cancer Center. All samples were extracted, cleaned-up, and analyzed using GC/MS. In the healthy person's samples, among the OCP compounds, the highest concentration was found for heptachlor-oxo-epoxide (5696.71 µg/kg), while among the PCB congeners, the non-ortho PCB 126 shows the highest TEQ concentrations (5554.5 µg TEQ/kg) and among the PCDDs/Fs congeners, the highest TEQ value was found for the congener 2,3,4,7,8-PeCDFs (5.93 µg TEQ/kg). For the cancer-affected patient's samples, the highest concentration among the OCP compounds was found for o,p-DDT (638.7 µg/kg), while among the PCBs congeners, the highest TEQ value was found for the non-ortho-PCB 126 (3366.24 µg TEQ/kg) and among the PCDDs/Fs congeners, the highest TEQ value was found for the congener 1,2,3,7,8-PeCDDs (20.64 µg TEQ/kg). OCP concentration level in adipose tissue samples for healthy people was 32 times higher than for cancer patient persons, while the TEQ values for dioxin-like PCB concentrations in adipose tissue samples of healthy people was 2.2 times higher than in the samples of cancer-affected patient and the TEQ values for PCDDs/Fs in adipose tissue samples of cancer-affected patient was 3 times higher than in the samples of healthy people.

Fatty acids, polychlorinated dibenzo-p-dioxins and dibenzofurans, and dioxin-like polychlorinated biphenyls in paired muscle and skin from fish from the Bohai coast, China: Benefits and risks associated with fish consumption.

Fish consumption benefits early cognitive development and cardiovascular health because of the n-3 long-chain polyunsaturated fatty acids in the fish, but toxic pollutants in fish, like dioxin-like polychlorinated biphenyls (dl-PCBs) and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), may decrease or counteract these benefits. In this study, the fatty acids, dl-PCBs, and PCDD/Fs were analyzed in paired muscle and skin from 13 fish and one squid species from the Bohai coast, which have been influenced by serious dioxin pollution from the Bohai Rim Region. The total fatty acid concentrations in the muscle and skin were 2.6-87 and 3.6-156 mg/g wet weight (ww), respectively. The total polyunsaturated fatty acid concentrations were higher in skin than muscle for almost half of the species. The total PCDD/F and dl-PCB toxic equivalents in the muscle and skin were 0.055-0.68 and 0.0099-0.43 pg/g ww, respectively, and were up to five times higher in the muscle than in the skin for eight species. Few benefit-risk studies for fish consumption with and without skin have been performed, so benefit-risk quotients (BRQs) for eating only muscle, only skin, and eating both were calculated. To achieve the recommended 250 mg of EPA + DHA intake, eating only muscle, only skin, and eating both from seven species with BRQs < 1 would cause no significant risk. Removing skin before eating is not necessary for these species. Three fish species had BRQs < 1 for muscle only but BRQs > 1 for skin only. Removing skin before eating would be better for these species. The other four fish species had BRQs > 1 for eating only muscle, only skin, and eating both, indicating the potential risk caused by PCDD/F and dl-PCB. Amounts of the four fish species consumed should be decreased, and other sources of EPA and DHA should be selected.

Association of prenatal exposure to PCDD/Fs and PCBs with maternal and infant thyroid hormones: The Hokkaido Study on Environment and Children's Health.

Polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and biphenyls (PCBs) are persistent organic pollutants that are universally detected. Some congeners of PCDDs, PCDFs or PCBs have dioxin-like toxicity, whereas non-dioxin-like PCBs are considered to have different toxicity. Reports of the relationships between prenatal exposure to PCDDs, PCDFs or PCBs and thyroid homeostasis in pregnant women and infants have been inconsistent. The aim of this study was to investigate the effect of maternal serum PCDD/F or PCB levels on maternal and neonatal thyroid hormone (TH) levels in a prospective cohort. Of the 514 subjects in the prospective cohort, 386 mothers and 410 infants were included for analysis. Fifteen dioxins and seventy PCBs in maternal blood collected between 23 and 41weeks of gestation were measured using high-resolution gas chromatography and high-resolution mass spectrometry. Blood samples to measure thyroid stimulating hormone (TSH) and free thyroxine (FT4) levels were obtained from mothers at an early gestational stage (median ten weeks), and from infants between four and seven days of age, respectively. Multiple linear regression analysis was conducted. Median concentration of total PCBs, PCB 153 were 104,700, and 20,500pg/g lipid, respectively. Median total dioxin-TEQ was 13.8pg/g lipid. Total dioxin-TEQ, coplanar PCBs were positively associated with neonatal FT4 (beta=0.224, 0.206, respectively). The association was stronger in boys (beta=0.299, 0.282, respectively). Several PCDD/F and PCB isomers were also positively associated with neonatal FT4. Total PCBs or non-dioxin-like PCBs were not associated with any maternal or neonatal THs. No DLC grouping or congeners were associated with neonatal TSH. Non-ortho PCBs were positively associated with maternal FT4. Three PCB congeners had significant positive association(s) with maternal THs. In conclusion, the results of our study suggest that perinatal exposure to background-level DLCs increases neonatal FT4, especially in boys.

Site-specific bioaccumulation of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/PCDFs) in mothers and their infants living in vicinity of Bien Hoa airbase, Southern Vietnam.

Bien Hoa airbase is located in Dong Nai Province (Southern Vietnam). Several sites within the airbase are highly contaminated by Agent Orange/Dioxin, and thus, they are also commonly named as an Agent Orange/Dioxin hotpot. In the present study, 36 maternal milk samples were collected from primiparas who have lived at least 5 years in four wards, Buu Long, Quang Vinh, Trung Dung and Tan Phong, which are closed to Bien Hoa airbase in order to investigate the level and distribution of Agent Orange/Dioxin exposure in different local communities. The mean concentrations of PCDD/PCDFs in four investigated wards range from 6.4 to 13.6 pgTEQ/g lipid. The highest mean TEQ of PCDD/PCDFs was observed in Buu Long ward (13.6 pgTEQ/g lipid), followed by Tan Phong ward (12.3 pgTEQ/g lipid), and the lowest value was observed in Trung Dung ward (6.4 pgTEQ/g lipid). The mean concentration of 2,3,7,8-TCDD in Buu Long (7.6 pg/g lipid) was approximately 2-6 times higher than those in Tan Phong (3.9 pg/g lipid), Quang Vinh (2.3 pg/g lipid), or Trung Dung (1.2 pg/g lipid). These results imply site-specific exposure to PCDD/PCDFs in different local communities living around Bien Hoa airbase. The mean values of daily intake of dioxin estimated for the breast fed infants living in Buu Long, Quang Vinh, Trung Dung and Tan Phong were about 80, 37.5, 31.7 and 58 pg TEQ/kg bw/day, respectively.

Seasonal variations of PCDD/Fs in fishes: inferring a hidden exposure route from Na-PCP application for schistosomiasis control.

Japanese schistosomiasis was endemic in Dongting Lake, the second largest freshwater lake in China. The technical sodium pentachlorophenate (Na-PCP) was widely used during the period between the early 1960s and early 1990s to kill oncomelania, the intermediate host of Schistosomes. However, polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) also contaminated the environment of Dongting Lake due to trace impurities of Na-PCP. In this study, 13 species of freshwater fish were collected from Dongting Lake and analyzed in wet and dry seasons. Temporal (wet and dry season) variations in PCDD/Fs in fishes from Dongting Lake were observed. The concentration of PCDD/Fs was considerably higher in the wet season than in the dry season and in areas in China where Na-PCP was not applied. The higher level of PCDD/Fs in the wet season may be attributed to the use of Na-PCP in the past and to the increase in the migration of PCDD/Fs from sediments to water in the wet season. The results indicate that the risk of PCDD/Fs contamination is connected with prior consumption of fishes that live in the Na-PCP application area. However, there was no difference in the concentrations of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) between the wet and dry seasons.

Association of dioxin exposure and reproductive hormone levels in men living near the Bien Hoa airbase, Vietnam.

Dioxins are endocrine-disrupting chemicals, and their effects on reproductive functions are well-documented. The aim of the present study was to measure the levels of reproductive hormones in 42 men residing near a dioxin-contaminated area in Vietnam. We measured levels of 17 2,3,7,8-substituted congeners of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) and four non-ortho polychlorinated biphenyls (PCBs) in blood. Levels of follicle-stimulating hormone, luteinizing hormone, progesterone, prolactin, estradiol, and total testosterone were measured in serum. Blood dioxin levels were elevated; the levels of 2,3,7,8-tetrachlorodibenzo-p-dioxin was 7.3pg/g fat. Seven of the men had testosterone levels below 250ng/dL, and nine men had prolactin levels above 9.7ng/mL. Four PCDD congeners, two PCDF congeners, one PCB congener, and the sum TEQ of PCDDs, PCDDs/Fs, and PCDDs/Fs/PCBs were positively and significantly correlated with prolactin levels. Two PCDD congeners, six PCDF congeners, two PCB congeners, and the TEQs of PCDFs and PCBs were negatively and significantly correlated with testosterone levels. There were no significant correlations between dioxin congeners and follicle-stimulating hormone, luteinizing hormone, or progesterone levels.

Persistent organic pollutants in krill from the Bellingshausen, South Scotia, and Weddell Seas.

Persistent organic pollutants (POPs) reach Antarctica through atmospheric transport, oceanic currents, and to minor extent, by migratory animals. The Southern Ocean is a net sink for many POPs, with a key contribution of the settling fluxes of POPs bound to organic matter (biological pump). However, little is known about POP transfer through the food web in the Southern Ocean and Antarctic waters, where krill is an important ecological node. In this study, we assessed the occurrence of polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs), polybrominated diphenyl ethers (PBDEs), and polychlorinated biphenyls (PCBs) in Antarctic krill (Euphausia superba) from the Bellingshausen, South Scotia and Weddell Seas around the Antarctic Peninsula. The concentrations of PCDD/Fs, PBDEs and PCBs in krill showed a large variability and the average were higher (generally within a factor 3) than those previously reported for eastern Antarctica. This result highlights regional differences related to atmospheric transport and deposition, and also probable regional sources due to human activities. Bioaccumulation and biomagnification factors for PCBs in krill were estimated using previously reported phytoplankton and seawater concentrations for this region. These suggested a near water-krill equilibrium for PCBs, which was not observed for water-phytoplankton partitioning. The estimated removal settling fluxes of PCBs due to the biological pump were several orders of magnitude higher than the estimated fluxes of PCBs transferred from phytoplankton to krill.

WHO/UNEP global surveys of PCDDs, PCDFs, PCBs and DDTs in human milk and benefit-risk evaluation of breastfeeding.

Since 1987, the World Health Organization (WHO) carried out global surveys on polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) in human milk. This study presents a review of the three most recent surveys from 2000 to 2010, including DDT. The objective was to identify global quantitative differences and provide baseline information for 52 countries or provide time-trends for countries with previous data. Individual human milk samples were collected following a WHO-designed procedure and combined to form a national pooled sample. Here, we report global levels for PCDDs, PCDFs, PCBs and the sum of o,p'-DDT, p,p'-DDT, o,p'-DDE, p,p'-DDE, o,p'-DDD and p,p'-DDD (ΣDDTs). A concise risk-benefit evaluation related to human milk contamination with these persistent organic pollutants (POPs) was also done. Large global and regional differences were observed. Levels of PCDDs and PCDFs were highest in India and some European and African countries. PCB levels were highest in East and West Europe. The highest levels of ΣDDTs were found in less industrialized countries. A temporal downward trend for PCDDs, PCDFs and PCBs is indicated. A risk-benefit assessment indicates that human milk levels of PCDDs, PCDFs and PCBs are still significantly above those considered toxicologically safe, while ΣDDTs are below or around those considered safe. With respect to potential adverse health effects, a more dominant role of in utero exposure versus lactational exposure is indicated. If potential adverse effects are balanced against positive health aspects for (breastfed) infants, the advantages of breastfeeding far outweigh the possible disadvantages. Our observations provide a strong argument to plea for further global source-directed measures to reduce human exposure further to dioxin-like compounds.

The influence of the Cucurbitaceae on mitigating the phytotoxicity and PCDD/PCDF content of soil amended with sewage sludge.

The study evaluates the impact of sewage sludge on OECD - Organization for Economic Cooperation and Development and vegetable soil phytotoxicity, measured using three test species: Lepidium sativum, Sinapis alba and Sorghum saccharatum, and total and TEQ PCDD/PCDF (toxic equivalency polychlorinated dibenzo-p-dioxins/polychlorinated dibenzofurans) soil concentration, measured using HRGC/HRMS - High Resolution Gas Chromatography/High Resolution Mass Spectrometry. It also evaluates the effect of zucchini and cucumber cultivation during 5-weeks period on mitigating these parameters. The application of 3, 9 and 18 t/ha of sewage sludge gradually increases the phytotoxicity of both OECD and vegetable soil. In the case of OECD soil, the highest roots growth inhibitions were observed for S. alba (73%, 86% and 87%, respectively) and the lowest for S. saccharatum (7%, 59% and 70%), while in vegetable soil inhibitions were averagely 25% lower. Sludge application also led to a 38% (3 t/ha), 169% (9 t/ha) and 506% (18 t/ha) increase in PCDD/PCDF concentration, and the TEQs were augmented by 15%, 159% and 251%. Both soil phytotoxicity and total and TEQ PCDD/PCDF concentrations were diminished as a result of zucchini and cucumber cultivation. The maximum reduction of soil phytotoxicity (83%) was observed as an effect of cucumber cultivation, while zucchini was 11% less effective. Zucchini, in turn, was more efficient in PCDD/PCDF removal (37% reduction), followed by cucumber (24%). Such differences were not observed in the case of TEQ reductions (68% and 66% for zucchini and cucumber cultivation, respectively).

Uptake and bioaccumulation of PCDD/Fs in earthworms after in situ and in vitro exposure to soil from a contaminated sawmill site.

Uptake of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) was studied in earthworms collected from a sawmill site in Sweden with severe PCDD/Fs contamination (the hot spot concentration was 690,000ng TEQWHO2005/kg d.w.) in order to investigate the transfer of PCDD/Fs from the site to the biota. PCDD/Fs concentrations in the collected earthworms were compared to PCDD/Fs concentrations in laboratory exposed earthworms (Eisenia fetida), which were exposed to contaminated soils from the sawmill site for 34days. All analyses were performed by high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS). PCDD/Fs concentrations in the earthworms ranged from 290 to 520,000pg/g (f.w.). The main congeners found in both soils and earthworms were OCDF, 1234678-HpCDF, OCDD and 1234678-HpCDD. The study showed that the PCDD/Fs in the soil were biovailable to the earthworms and the PCDD/Fs concentrations in the soils correlated with the concentrations in the earthworms. Earthworm samples from soil with lower concentration had higher bioaccumulation factors than samples from soils with high concentration of contamination. Thus, a less contaminated soil could yield higher concentrations in earthworms compared to a higher contaminated soil. Assuming that when assessing risks with PCDD/F contaminated soil, a combination of chemical analysis of soil PCDD/Fs concentrations and bioavailability should be employed for a more comprehensive risk assessment.

Influence of trophic ecology on the accumulation of dioxins and furans (PCDD/Fs), non-ortho polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) in Mediterranean gulls (Larus michahellis and L. audouinii): A three-isotope approach.

The impact of pollution caused by severe anthropogenic pressure in the Mediterranean Sea, an important biodiversity hotspot, requires continuous research efforts. Sources of highly toxic chemicals such as Persistent Organic Pollutants (POPs) are misunderstood in representative Mediterranean species, which limits our capability to establish proper conservation strategies. In the present study, eggs of Audouin's and yellow-legged gulls (Larus audouinii and L. michahellis) were used to investigate the trophic sources, as measured by δ(13)C, δ(15)N, and δ(34)S, of legacy POPs, in particular, polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs) and non-ortho polychlorinated biphenyls (no-PCBs), as well as recently-regulated POPs, e.g., polybrominated diphenyl ethers (PBDEs). Special attention was paid to the usefulness of rarely-explored δ(34)S ratios in explaining POP exposure in wildlife, and δ(34)S was the isotopic ratio that best explained POP variations among gulls in most cases, thus demonstrating its usefulness for understanding POP exposure in wildlife. Significant relationships between stable isotope signatures and POP concentrations revealed increasing levels of no-PCBs and low halogenated PCDD/Fs and PBDEs in Mediterranean gulls as the consumption of marine resources increases. In contrast, highly chlorinated and brominated congeners appeared to preferentially accumulate in gulls feeding primarily on refuse from dump sites and terrestrial food webs. The use of suitable dietary tracers in the study of POPs in yellow-legged gulls revealed the importance of dump sites as a source of POPs in Mediterranean seabirds, which has not previously been reported. In contrast, the preferential accumulation through marine food webs of low chlorinated PCCD/Fs and no-PCBs, which show the highest toxic equivalents factors (TEFs), led to a significantly greater toxicological concern in Audouin's as compared to yellow-legged gulls. Audouin's gull exposure to POPs appears primarily related to the pelagic food webs commonly exploited by fisheries, highlighting the need for further research given the potential impact on human consumption.