Explaining the impact of mutations on quantification of SARS-CoV-2 in wastewater.

Wastewater surveillance is an effective tool for monitoring community spread of COVID-19 and other diseases. Quantitative PCR (qPCR) analysis for wastewater surveillance is more susceptible to mutations in target genome regions than binary PCR analysis for clinical surveillance. The SARS-CoV-2 concentrations in wastewater estimated by N1 and N2 qPCR assays started to diverge around July 2022 in data from different sampling sites, analytical methods, and analytical laboratories in Japan. On the basis of clinical genomic surveillance data and experimental data, we demonstrate that the divergence is due to two mutations in the N1 probe region, which can cause underestimation of viral concentrations. We further show that this inaccuracy can be alleviated if the qPCR data are analyzed with the second derivative method or the Cy0 method instead of the crossing point method.

Liquid chromatography-electrospray ionization-tandem mass spectrometry for the determination of short-chain chlorinated paraffins in mixed plastic wastes.

Wastes containing short-chain chlorinated paraffins (SCCPs) at concentrations above the Basel Convention low persistent organic pollutant content (LPC) values must be destroyed or irreversibly transformed in an environmentally-sound manner. Here, we developed a novel liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MSMS) method for determining the concentrations of SCCPs in mixed plastic wastes. Major SCCP homologues were identified with good separation and peak width by using a low-hydrophobicity cyano-propyl column and a mobile phase consisting of water and methanol containing ammonium acetate. Precursor ion peaks corresponding to the formation of stable SCCP homologue-adducts were observed, followed by two intense product ion peaks corresponding to separation of the adduct into acetate and the homologue. The formulation of a novel calibration standard with known SCCP homologue percentage composition supported the development of our novel mass spectrometric technique. The results obtained with the LC-ESI-MSMS system were quantitatively and qualitatively comparable with those obtained with a high-resolution mass spectrometry (HRMS) coupled to gas chromatography (GC) system. Homologue concentrations determined by LC-ESI-MSMS were significantly correlated with those determined by GC-HRMS in samples of commercial chlorinated paraffin mixture and mixed plastic waste, respectively. As a complementary technique to the highly accurate, but less versatile GC-HRMS approaches, the SCCP analysis by LC-ESI-MSMS is a practical way to identify mixed plastic wastes containing SCCPs at concentrations higher than the Basel Convention's LPC value.

PCDD/PCDF formation in the chlor-alkali process-laboratory study and comparison with patterns from contaminated sites.

Studies on the formation of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) during the electrolysis of sodium chloride solution (brine) using graphite or titanium electrodes were carried out at a laboratory scale. High concentrations of PCDFs but no PCDDs were formed in tests using graphite electrodes. With titanium electrodes, PCDFs were only formed when tar pitch was added and mainly originated from the dibenzofuran present in the tar. For the first time, a detailed assessment of the formation of mono- to octachlorinated PCDD/PCDF from tar pitch was investigated. The assessment included of the chlorination steps proved that PCDFs were formed by successive lateral chlorinated from dibenzofuran to MonoCDFs, DiCDFs, and TriCDFs to form the typical known "chlorine pattern" of TetraCDF to OctaCDF with a dominance of 1,2,7,8- and 2,3,7,8-TetraCDFs, 1,2,3,7,8-PentaCDF, and 1,2,3,4,7,8-HexaCDF as marker congeners. The final homologue distributions depended on reaction time and reaction temperature. In addition, electrolysis with non-chlorinated dibenzo-p-dioxins, dibenzofuran, and biphenyl was carried out. As a result, PCDDs, PCDFs, and PCB were formed at comparable yields. Congener patterns in soil samples from a PCDD/F-contaminated site where chlor-alkali electrolysis had been operated for decades in Japan had identical isomer distribution demonstrating the source and contamination potential and risk of these processes. Therefore, sites where in the past 120 years chlor-alkali electrolysis has been operated or where residues from chlor-alkali production or other chlorine using industries have been disposed should be assessed for their pollution level and exposure relevance. The assessment of total organohalogen content revealed that PCDF is only a small fraction of organohalogens in the contaminated soils. For an appropriate risk assessment, also other chlorinated aromatic compounds such as PCBs or PCNs need to be considered.

Associations between levels of hydroxylated PCBs and PCBs in serum of pregnant women and blood thyroid hormone levels and body size of neonates.

The purpose of this study was to investigate possible associations between concentrations of hydroxylated PCBs (OH-PCBs) and PCBs in the serum of women in the first trimester of pregnancy and thyroid hormone levels and body size of newborn infants in 79 mother-neonate pairs. We measured 16 OH-PCB isomers and 29 PCB isomers in the serum of Japanese women sampled at 11.1±1.9 weeks of gestation. The concentrations of free thyroxine (fT4) and thyroid stimulating hormone (TSH) were measured in whole blood spots on filter papers sampled from the neonates. Dietary and lifestyle information of the mothers were obtained by self-administered questionnaires. Geometric mean (GM) concentrations of the sum of 16 OH-PCB isomers and of 29 PCB isomers were 1.2×10(2)pg/g wet wt. and 69ng/g lipid wt., respectively, in maternal serum. The GM concentrations of neonatal fT4 and TSH were 2.21ng/dL and 1.37µIU/mL, respectively. Multiple regression analysis was performed using measures of neonatal thyroid hormones as dependent variable and serum levels of OH-PCBs/PCBs and other potential covariates (age, pre-pregnancy weight, smoking status, etc.) as independent variables. The results demonstrated a significant positive association between the concentrations of some OH-PCB isomers and that of neonatal TSH. There were no significant associations between levels of PCBs and neonatal fT4, or between OH-PCBs/PCBs and body size of neonates. We conclude that exposure to/body burden of OH-PCBs, but not PCBs, at environmental levels during the first trimester of pregnancy can affect neonatal thyroid hormone status.

Serum levels of hydroxylated PCBs, PCBs and thyroid hormone measures of Japanese pregnant women.

OBJECTIVES: The purpose of this study was to investigate the associations between serum concentrations of hydroxylated PCBs (OH-PCBs) and PCBs and measures of thyroid hormone status of Japanese pregnant women. METHODS: The concentrations of free thyroxine (fT4), thyroid stimulating hormone (TSH), and thyroxine binding globulin (TBG) as well as 16 OH-PCB isomers and 29 PCB isomers were analyzed in the serum of 129 women sampled in the first trimester of gestation. Dietary and lifestyle information of the subjects was obtained by self-administered questionnaire. Multiple regression analysis was performed using measures of thyroid hormones as the dependent variable and serum levels of OH-PCBs/PCBs, urinary iodine concentration, and other potential covariates (age, BMI, smoking, etc.) as independent variables. RESULTS: Geometric mean (GM) concentration of the sum of 16 isomers of OH-PCBs was 120 pg/g wet wt. and that of 29 isomers of PCBs was 68 ng/g lipid wt., respectively, in the serum of the subjects. Iodine nutrition was considered adequate to high from urinary iodine level (GM, 370 µg/g creatinine). The mean concentration of TSH, fT4 and TBG was 1.34 ± 1.37 µIU/mL, 1.22 ± 0.16 ng/dL and 33.0 ± 6.4 µg/mL, respectively, with a small number of subjects who were outside the reference range. Multiple regression analysis revealed that serum concentrations of OH-PCBs/PCBs were not significantly associated with any of the measures of thyroid hormone status. CONCLUSIONS: Exposure/body burden of OH-PCBs and PCBs at environmental levels does not have a measurable effect on thyroid hormones.

Analysis of UNEP priority POPs using HRGC-HRMS and their contamination profiles in livers and eggs of great cormorants (Phalacrocorax carbo) from Japan.

The present investigation demonstrates establishment of United Nations Environment Programme (UNEP) priority Persistent Organic Pollutants (POPs) using high-resolution gas chromatography-high-resolution mass spectrometry. Particularly, POPs analytical methods were established using native and (13)C-labeled internal standards of HCHs, HCB, cyclodienes, chlordanes, DDTs, mirex, dioxin-like PCBs, PCDDs, and PCDFs by isotope dilution technique. The relative response factor for 6-point calibration curve native standards (18 replicate analysis) were in the ranges of 0.93-1.43 with relative standard deviation ranges from 1.68 to 4.96%. Instrument detection limit and instrument quantification limit was established for various POPs at femtograms. Concentrations of UNEP-POPs were measured in liver (n = 10) and egg (n = 10) of great cormorants and their major diet, gizzard shad (n = 2), collected in and around Tokyo, Japan. DDTs (ranges in liver and egg, respectively) were predominant accumulants (9800-310,000 and 9600-73,000) followed by dioxin-like PCBs (4500-69,000 and 7900-150,000), chlordanes (2600-16,000 and 700-4,800), cyclodienes (650-4600 and <1-1000), HCB (680-2800 and 180-590), HCHs (230-1800 and 120-490), PCDD/DFs (3.2-27 and 1.7-5.7) on nanogram per gram lipid basis. Concentrations (ranges) of POPs in gizzard shad were in the following order: DDTs (3900-16,000), chlordanes (3400-14,000), cyclodienes (340-1300), HCB (110-480), and HCHs (140-360) on nanogram per gram lipid basis.

Impact of FEBRA (fermented brown rice with Aspergillus oryzae) intake and concentrations of PCDDs, PCDFs and PCBs in blood of humans from Japan.

Impact of FEBRA (fermented brown rice with Aspergillus oryzae, a rich dietary fiber) intake on the concentrations of polychlorinated-biphenyls (PCBs), -dioxin-like polychlorinated biphenyls (DLPCBs), -dibenzo-p-dioxins (PCDDs), and -dibenzofurans (PCDFs) in blood of nine married Japanese were studied for 2 years. Concentrations of total PCBs (average+/-standard deviation) were greater 326,000+/-263,000 and 207,000+/-89,000 in FEBRA-intake and non-FEBRA-intake groups, respectively, on pg/g lipid (from now onwards data expressed on pg/g lipid). DLPCBs were second major contaminants 94,000+/-77,000 and 60,000+/-28,000, orderly in FEBRA-intake and non-FEBRA-intake groups. The concentrations of PCDD/DFs were several orders of magnitude lower than those of PCBs, whereas PCDDs were greater (433+/-233 and 512+/-511, in FEBRA-intake and non-FEBRA-intake groups, respectively) than PCDFs (56+/-50 and 43+/-35). Calculated toxic equivalency (TEQ) concentrations were 34+/-24 and 23+/-10 in FEBRA-intake and non-FEBRA-intake groups, respectively. In order to corroborate the impact of FEBRA, the changes of PCDD/DFs and PCB concentrations for 0.5-year, 1-year, 1.5-year, and 2-year samples were estimated from average concentration of 1-day before study commences and 1-week study samples. The results explicit FEBRA-intake group eliminated 7%, 33%, 45%, 36%, and 26% and non-FEBRA-intake group eliminated 8%, 28%, 36%, 31%, and 20% of DLPCBs, PCDDs, PCDFs, PCDD/DF TEQ and total TEQ, respectively. This study is the first of its kind showing the possible elimination of PCDDs, PCDFs and their TEQ in humans from Japan after the consumption of FEBRA.

Impact of fermented brown rice with Aspergillus oryzae (FEBRA) intake and concentrations of polybrominated diphenylethers (PBDEs) in blood of humans from Japan.

The isotope dilution technique was applied for the analysis of new polybrominated diphenylethers (PBDEs) calibration standard (both labeled and non-labeled) using high-resolution gas chromatography-high-resolution mass spectrometry (HRGC-HRMS). The relative response factor (RRF) and relative standard deviation (RSD) for new calibration standard in Finnigan Thermo Electron (MAT-95XL) and Micromass (Autospec Ultima) were more or less identical with mean RRF (0.9882), RSD (0.0865) and CV% of (8.75). The results also revealed for DeBDE-209 quantification; labeled DeBDE-209 is essential. Furthermore, we recommend on column injection technique with a thin film instead of splitless injection in order to reduce the thermal degradation of DeBDE-209 and formation of octabromodibenzofurans (OBDF). Besides, analysis of human blood (n = 156) of FEBRA-intake and non-FEBRA-intake individuals elucidated frequent detection of eighteen PBDE congeners. The average PBDE concentrations in non-FEBRA intake and FEBRA-intake humans were 6000-11,000 (mean: 8400) and 5400-15,000 (mean: 9900) respectively on pg/g fat basis. Although FEBRA-intake individuals showed slightly greater PBDEs, computer-normalized concentrations of TeBDE-47 corroborate FEBRA-intake individual from four family showed reduced concentrations. The contamination profiles of PBDEs varied in between family, gender as well as geography. International comparison with predominant PBDE congener (TeBDE-47) prevailed lower levels in Japan when compared to Korea, Germany and USA nevertheless, congener specific profiles were different which is in accordance with different technical PBDE usage in between countries.