Heavy metal levels and human health risk implications associated with fish consumption from the lower Omo river (Lotic) and Omo delta lake (Lentic), Ethiopia.

This study is the first to determine the levels of heavy metals in commercially important fish species, namely Lates niloticus and Oreochromis niloticus and the potential human health risks associated with their consumption. A total of 120 fish samples were collected from the lower Omo river and Omo delta, with 60 samples from each water source. The fish tissue samples (liver and muscle) were analyzed using a flame atomic absorption spectrometer for nine heavy metals (Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn). The human health risk assessment tools used were the target hazard quotient (THQ), the hazard index (HI), and the target cancer risk (TCR). The mean levels of heavy metals detected in the liver and muscle of L. niloticus from the lower Omo river generally occurred in the order Fe > Zn > Pb> Cu > Mn> Cr > Co > Ni and Pb > Cu > Mn > Co > Ni, respectively. The mean levels of metals in the muscle and liver tissues of O. niloticus were in the order Fe > Pb > Zn > Mn > Cu > Cr > Co > Ni and Pb > Zn > Mn > Fe > Cu > Co > Ni, respectively. Similarly, the mean levels of heavy metals detected in the liver and muscle of L. niloticus from Omo delta occurred in the order Fe > Zn > Pb > Cu > Mn > Cr > Co > Ni and Fe > Pb > Zn > Mn > Cu > Co > Cr > Ni, respectively. The mean levels in the muscle and liver tissues of O. niloticus from the Omo delta were in the order Fe > Pb > Zn > Mn > Cu > Cr > Co > Ni and Pb > Fe > Zn > Mn > Co > Cu > Ni, respectively. The study revealed that the THQ values were below 1, indicating that consumption of L. niloticus and O. niloticus from the studied sites does not pose a potential non-carcinogenic health risk. Although the TCR values for Pb in this study were within the tolerable range, it's mean concentration in the muscle and liver tissues of both fish species from the two water bodies exceeded the permissible limit established by FAO/WHO. This is a warning sign for early intervention, and it emphasizes the need for regular monitoring of freshwater fish. Therefore, it is imperative to investigate the pollution levels and human health risks of heavy metals in fish tissues from lower Omo river and Omo delta for environmental and public health concerns.

Ocimum gratissimum essential oil in the transport water of Brycon hilarii: implications at water quality, blood parameters and residues in tissue and plasma.

Transporting live fish is a common practice in fish farming, and is certainly one of the main problems that affect fish homeostasis. In this scenario, the use of natural additives has shown promise in improving fish resistance to adverse situations. This study aimed to assess the impact of Ocimum gratissimum L. essential oil (OGEO) on water quality, hematological parameters, and residue levels in the plasma, fillet, and liver of juvenile piraputanga (Brycon hilarii) during a two-hour transportation period. The fish were divided into plastic bags (4 L) and exposed to three different OGEO concentrations (10, 20, and 30 mg L-1), while a control group received no OGEO (three repetitions each). After the two-hour transportation, blood samples were collected, as well as portions of the fillet and liver for quantifying essential oil compounds, which were also measured in the plasma. Oxygen levels remained high throughout the transportation period, in all groups, while the pH decreased. Hemoglobin, MCHC, and MCH increased in fish exposed to OGEO concentrations of 20 and 30 mg L-1, compared to the control group. However, lymphocyte counts and the concentrations of essential oil compounds in plasma, fillet, and liver increased with higher OGEO concentrations. The use of 10 mg L-1 OGEO in the two-hour transport water is promising to ensure the survival and well-being of Brycon hilarii juveniles (weighing 16 g), showing to be safe and effective. The residual concentration of eugenol the major compound of OGEO in the fillet remains below the maximum limit of the recommended daily intake.

The red listed eagle owl (Bubo bubo) population in Norway is exposed to POP levels exceeding threshold values for adverse health effects.

The eagle owl (Bubo bubo) population in Norway is today classified as critically endangered on the red list of endangered species. Because previous studies have detected high concentrations of Persistent Organic Pollutants (POPs) in birds of prey, concerns have been raised whether POPs exposure are a significant factor to the substantial decline of the eagle owl population. The aims of this study were to measure the levels of POPs in eagle owls and to assess whether POPs may represent a potential health risk. POPs were analysed in liver samples from 100 eagle owls collected between 1994 and 2014. The concentrations of POPs were generally very high and individual birds had levels among the highest measured worldwide. The contaminant groups analysed were highly correlated (p < 0.0001). The concentrations of sum of Polychlorinated Biphenyls (∑PCB) exceeded the threshold value from moderate to severe health risk in 90% of the birds. The birds with cachectic or lean body condition had significantly higher levels of contaminants than those with higher body condition scores. No significant temporal or spatial trends were noted. The lack of temporal trends, suggest that the downward trend of POPs, appear to be levelling off. The lack of differences between inland and coastal regions suggest that the risk of exposure may be comparable between predatory birds feeding in marine or terrestrial food webs. The significantly higher POPs levels detected in individuals with poor body condition may be due to reduced fat stores and thereby higher concentration in the remaining fat and/or the weight loss could be induced by toxic effects. The high proportion of birds exceeding the threshold values for severe and high risk of adverse effects, suggest that the high contamination load may reduce the eagle owl's fitness and survival and, thus, contribute to decline of the eagle owl population.

Changes in the amount of nucleotide sugars in aged mouse tissues.

Aging affects tissue glycan profiles, which may alter cellular functions and increase the risk of age-related diseases. Glycans are biosynthesized by glycosyltransferases using the corresponding nucleotide sugar, and the availability of nucleotide sugars affects glycosylation efficiency. However, the effects of aging on nucleotide sugar profiles and contents are yet to be elucidated. Therefore, this study aimed to investigate the effects of aging on nucleotide sugars using a new LC-MS/MS method. Specifically, the new method was used to determine the nucleotide sugar contents of various tissues (brain, liver, heart, skeletal muscle, kidney, lung, and colon) of male C57BL/6NCr mice (7- or 26-month-old). Characteristic age-associated nucleotide sugar changes were observed in each tissue sample. Particularly, there was a significant decrease in UDP-glucuronic acid content in the kidney of aged mice and a decrease in the contents of several nucleotide sugars, including UDP-N-acetylgalactosamine, in the brain of aged mice. Additionally, there were variations in nucleotide sugar profiles among the tissues examined regardless of the age. The kidneys had the highest concentration of UDP-glucuronic acid among the seven tissues. In contrast, the skeletal muscle had the lowest concentration of total nucleotide sugars among the tissues; however, CMP-N-acetylneuraminic acid and CDP-ribitol were relatively enriched. Conclusively, these findings may contribute to the understanding of the roles of glycans in tissue aging.

A high-throughput UPLC-MS-MS Bio-analytical method for the analysis of veterinary pharmaceutical residues in Chicken Tissues, Application of efficient-valid-green (EVG) Framework as a Competence Tool.

Antibacterial medications are receiving the most attention due to hypersensitivity reactions and the emergence of bacterial mutants resistant to antibiotics. Treating Animals with uncontrolled amounts of antibiotics will extend beyond their lives and affect humans. This study aims to determine the concentration of the residues of sulfadimidine, sulfaquinoxaline, diaveridine, and vitamin K3 in the tissues of poultry (muscles and liver) after treatment with the combined veterinary formulation. A UPLC-MS-MS method was developed using Poroshell 120 ECC18 and a mobile phase composed of acetonitrile and distilled water, containing 0.1 % formic acid, in the ratio of (85:15 v/v) at a flow rate of 0.6 mL/min. Sample extraction solvent was optimized using response surface methodology (RSM) to be acetonitrile: methanol in the ratio (49.8: 50.2 v/v), and the method was validated according to the FDA bioanalytical method validation protocol over the range (50-1000 µg/Kg) for sulfaquinoxaline and (50-750 µg/Kg) for the other 3 drugs. The greenness of the sample preparation and analytical method was assessed by applying Analytical Eco-scale (AES) and AGREE coupled with AGREEprep. The Competence of the study was evaluated via the EVG framework known as Efficiency, validation, and greenness, to achieve a balance point represented by a radar chart. The method was applied to decide the time required for poultry products to be safe for human use after administration of the studied drugs. It was found that, after the administration of the last dose, minimally 7 days are required till the levels of the drugs drop to the maximum residue limit determined by the FDA/WHO in animal tissues.

Development and validation of a GC-MS/MS method for the determination of iodoacetic acid in biological samples.

Iodoacetic acid (IAA) is a halogenated disinfection by-product of growing concern due to its high cytotoxicity, genotoxicity, endocrine disruptor effects, and potential carcinogenicity. However, the data on distribution and excretion of IAA after ingestion by mammals are still scarce. Here, we developed a reliable and validated method for detecting IAA in biological specimens (plasma, urine, feces, liver, kidney, and tissues) based on modified QuEChERS sample preparation combined with gas chromatography-tandem triple quadrupole mass spectrometry (GC-MS/MS). The detection method for IAA exhibited satisfactory recovery rates (62.6-108.0%) with low relative standard deviations (RSD < 12.3%) and a low detection limit for all biological matrices ranging from 0.007 to 0.032 ng/g. The study showed that the proposed method was reliable and reproducible for analyzing IAA in biological specimens. It was successfully used to detect IAA levels in biological samples from rats given gavage administration. The results indicated that IAA was found in various tissues and organs, including plasma, thyroid, the liver, the kidney, the spleen, gastrointestinal tract, and others, 6 h after exposure. This study provides the first data on the in vivo distribution in and excretion of IAA by mammals following oral exposure.

Visualization of PFOA accumulation and its effects on phospholipid in zebrafish liver by MALDI Imaging.

Exposure to poly- and perfluoroalkyl substances (PFASs) can result in bioaccumulation. Initial findings suggested that PFASs could accumulate in tissues rich in both phospholipids and proteins. However, our current understanding is limited to the average concentration of PFASs or phospholipid content across entire tissue matrices, leaving unresolved the spatial variations of lipid metabolism associated with PFOA in zebrafish tissue. To address gap, we developed a novel methodology for concurrent spatial profiling of perfluorooctanoic acid (PFOA) and individual phospholipids within zebrafish hepatic tissue sections, utilizing matrix-assisted laser desorption/ionization time of flight imaging mass spectrometry (MALDI-TOF-MSI). 5-diaminonapthalene (DAN) matrix and laser sensitivity of 50.0 were optimized for PFOA detection in MALDI-TOF-MSI analysis with high spatial resolution (25 µm). PFOA was observed to accumulate within zebrafish liver tissue. H&E staining results corroborating the damage inflicted by PFOA accumulation, consistent with MALDI MSI results. Significant up-regulation of 15 phospholipid species was observed in zebrafish groups exposed to PFOA, with these phospholipid demonstrating varied spatial distribution within the same tissue. Furthermore, co-localized imaging of distinct phospholipids and PFOA within identical tissue sections suggested there could be two distinct potential interactions between PFOA and phospholipids, which required further investigation. The MALDI-TOF-IMS provides a new tool to explore in situ spatial distributions and variations of the endogenous metabolites for the health risk assessment and ecotoxicology of emerging environmental pollutants.

Optimization of an HPLC-MS/MS method to analyze chlordecone in bovine serum and correlations with levels in liver, muscle and fat.

Chlordecone is an organochlorine pesticide used from 1972 to 1993 in the French West Indies. Its extensive use and high persistence in soils induced massive contamination of the environment and of the food chain, especially in cattle through contaminated soil ingestion. To ensure suitability for consumption of bovine meat, monitoring plans are set up based on perirenal fat concentrations after slaughtering. In the present study, we have investigated an in-vivo monitoring approach by measuring chlordecone levels in serum samples. For this purpose, a sensitive high-performance liquid-chromatography-tandem mass spectrometry (HPLC-MS/MS) method following a QuEChERS extraction method was successfully optimized and validated, reaching a limit of quantification of 0.05 ng g-1 fresh weight. This method was applied to 121 serum samples collected from bovines originating from contaminated areas of Martinique and Guadeloupe. Chlordecone was detected in 88% of the samples, and quantified in 77% of the samples, with concentrations ranging from 0.05 to 22 ng g-1. Perirenal fat, liver, and muscle were also sampled on the same animals and the measured concentrations of chlordecone were statistically correlated to the levels determined in serum. Mean concentration ratios of 6.5 for fat/serum, 27.5 for liver/serum, and 3.3 for muscle/serum were calculated, meaning that chlordecone was not only distribute in fat (as expected), muscle and liver, but also in serum. Good correlations were found to allow prediction of chlordecone concentrations in muscle based on concentrations measured in serum. This study opens the door to possible pre-control of bovines before slaughter. In cases of probable non-compliance with maximum residue levels (MRLs), farm management could proceed to allow for depuration under controlled conditions. This would have a strong impact on both economic and food safety management measures.

Lipid Wizard: Analysis Software for Comprehensive Two-Dimensional Liquid Chromatography-Mass Spectrometry-Based Lipid Profiling.

Lipids play a significant role in life activities and participate in the biological system through different pathways. Although comprehensive two-dimensional liquid chromatography-mass spectrometry (2DLC-MS) has been developed to profile lipid abundance changes, lipid identification and quantification from 2DLC-MS data remain a challenge. We created Lipid Wizard, open-source software for lipid assignment and isotopic peak stripping of the 2DLC-MS data. Lipid Wizard takes the peak list deconvoluted from the 2DLC-MS data as input and assigns each isotopic peak to the lipids recorded in the LIPID MAPS database by precursor ion m/z matching. The matched lipids are then filtered by the first-dimension retention time (1D RT), followed by the second-dimension retention time (2D RT), where the 2D RT of each lipid is predicted using an equivalent carbon number (ECN) model. The remaining assigned lipids are used for isotopic peak stripping via an iterative linear regression. The performance of Lipid Wizard was tested using a set of lipid standards and then applied to study the lipid changes in the livers of mice (fat-1) fed with alcohol.

Optimized 13C-TrEnDi Enhances the Sensitivity of Plasmenyl Ether Glycerophospholipids and Demonstrates Compatibility with Other Derivatization Strategies.

The identification and quantitation of plasmalogen glycerophospholipids is challenging due to their isobaric overlap with plasmanyl ether-linked glycerophospholipids, susceptibility to acid degradation, and their typically low abundance in biological samples. Trimethylation enhancement using diazomethane (TrEnDi) can be used to significantly enhance the signal of glycerophospholipids through the creation of quaternary ammonium groups producing fixed positive charges using 13C-diazomethane in complex lipid extracts. Although TrEnDi requires a strong acid for complete methylation, we report an optimized protocol using 10 mM HBF4 with the subsequent addition of a buffer solution that prevents acidic hydrolysis of plasmalogen species and enables the benefits of TrEnDi to be realized for this class of lipids. These optimized conditions were applied to aliquots of bovine liver extract (BLE) to achieve permethylation of plasmalogen lipids within a complex mixture. Treating aliquots of unmodified and TrEnDi-derivatized BLE samples with 80% formic acid and comparing their liquid chromatography mass spectrometry (LCMS) results to analogous samples not treated with formic acid, enabled the identification of 29 plasmalogen species. On average, methylated plasmalogen species from BLE demonstrated 2.81-fold and 28.1-fold sensitivity gains over unmodified counterparts for phosphatidylcholine and phosphatidylethanolamine plasmalogen species, respectively. Furthermore, the compatibility of employing 13C-TrEnDi and a previously reported iodoacetalization strategy was demonstrated to effectively identify plasmenyl-ether lipids in complex biological extracts at greater levels of sensitivity. Overall, we detail an optimized 13C-TrEnDi derivatization strategy that enables the analysis of plasmalogen glycerophospholipids with no undesired cleavage of radyl groups, boosting their sensitivity in LCMS and LCMS/MS analyses.

GenX caused liver injury and potential hepatocellular carcinoma of mice via drinking water even at environmental concentration.

Hexafluoropropylene oxide dimer acid (GenX) is an alternative to perfluorooctanoic acid (PFOA), whose environmental concentration is close to its maximum allowable value established by the US Environmental Protection Agency, so its effects on human health are of great concern. The liver is one of the most crucial target organ for GenX, but whether GenX exposure induces liver cancer still unclear. In this research project, male C57 mice were disposed to GenX in drinking water at environmental concentrations (0.1 and 10 µg/L) and higher concentrations (1 and 100 mg/L) for 14 weeks to explore its effects on liver injury and potential carcinogenicity in mice. GenX was found to cause a dose-dependent increase in the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), and triglyceride (TG). As the content of GenX in drinking water increased, so did the concentrations of Glypican-3 (GPC-3) and detachment gamma-carboxyprothrombin (DCP), indicators of early hepatocellular cancer. GenX destroyed the boundaries and arrangements of hepatocytes, in which monocyte infiltration, balloon-like transformation, and obvious lipid vacuoles were observed between cells. Following exposure to GenX, Masson sections revealed a significant quantity of collagen deposition in the liver. Alpha-feto protein (AFP), vascular endothelial growth factor (VEGF), Ki67, matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9) gene expression increased in a dose-dependent manner in the treatment group relative to the control group. In general, drinking water GenX exposure induced liver function impairment, elevated blood lipid level, caused liver pathological structure damage and liver fibrosis lesions, changed the liver inflammatory microenvironment, and increased the concentration of liver-related tumor indicator even in the environmental concentration, suggesting GenX is a potential carcinogen.

Persistent organic pollutants and polycyclic aromatic hydrocarbons in livers of stranded Arctocephalus australis in southern Brazilian beaches.

23 livers of South American fur seal (Arctocephalus australis) found stranded in southern Brazilian beaches were evaluated for Persistent Organic Pollutants (POPs) and Polycyclic Aromatic Hydrocarbons (PAHs). POPs (DDTs, mirex, eldrin, dieldrin, aldrin, isodrin, HCHs, chlordanes and PCBs) and PAHs in livers were Soxhlet extracted, analyzed and quantified using Gas Chromatography Tandem Mass Spectrometry (GC-TQMS). The main POPs found were PCBs and DDTs, totaling 81 %. Among pesticides, mirex followed DDTs, possibly due to usage in Uruguay, followed by Σdrins, ΣCHLs and ΣHCHs. Naphthalene was the major PAH found, while heavier compounds did not significantly bioaccumulate. Concentrations of POPs resembled previous findings for A. australis. Considering only juveniles, no POPs showed significant differences between sexes. Lipidic content, weight and length did not show any correlation with POP concentration. This was the first record of PAHs and PBDEs in South American fur seals, and the levels of these pollutants were relatively low.

Diazobutanone-assisted isobaric labelling of phospholipids and sulfated glycolipids enables multiplexed quantitative lipidomics using tandem mass spectrometry.

Mass spectrometry-based quantitative lipidomics is an emerging field aiming to uncover the intricate relationships between lipidomes and disease development. However, quantifying lipidomes comprehensively in a high-throughput manner remains challenging owing to the diverse lipid structures. Here we propose a diazobutanone-assisted isobaric labelling strategy as a rapid and robust platform for multiplexed quantitative lipidomics across a broad range of lipid classes, including various phospholipids and glycolipids. The diazobutanone reagent is designed to conjugate with phosphodiester or sulfate groups, while accommodating various functional groups on different lipid classes, enabling subsequent isobaric labelling for high-throughput multiplexed quantitation. Our method demonstrates excellent performance in terms of labelling efficiency, detection sensitivity, quantitative accuracy and broad applicability to various biological samples. Finally, we performed a six-plex quantification analysis of lipid extracts from lean and obese mouse livers. In total, we identified and quantified 246 phospholipids in a high-throughput manner, revealing lipidomic changes that may be associated with obesity in mice.

Exposure assessment of anticoagulant rodenticides in the liver of red foxes (Vulpes vulpes) in Slovenia.

The study deals with the environmental residues of anticoagulant rodenticides (ARs) in Slovenia to evaluate the toxicological risk of secondary poisoning of red foxes (Vulpes vulpes) as representatives of non-target wildlife, and in relation to the investigated use patterns of ARs and specific local parameters in Slovenia. From 2019 to 2022, 148 liver tissue samples of adult red foxes were collected from almost all state geographical regions. The samples were extracted with methanol/water (2:1, v/v), cleaned-up using a solid supported liquid-liquid extraction, and measured by liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS) with reporting limits of 0.5 to 5.0 ng/g. Residues of at least one rodenticide were detected in 77.7 % of the samples. The second generation ARs of bromadiolone, brodifacoum and difenacoum were the most frequently found, appearing in 75.0, 51.4, and 18.9 % of the samples, respectively. Concentrations of pooled ARs ranged from 1.5 to 2866.5 ng/g with mean and median values of 601.4 and 350.2 ng/g, respectively. We determined bromadiolone and brodifacoum at concentrations of ≥800 ng/g in 10.8 and 10.1 % of the samples, and 1.4 and 0.7 % of the samples contained residues >2000 ng/g, respectively. These concentrations are much higher than those found in comparable studies in Europe and elsewhere in the world. Residues of ARs were detected in all monitored statistical regions of Slovenia, with higher concentrations in the eastern parts of the country. First generation ARs were found in only 9.5 % of samples, and residues were below 10 ng/g with one exception (coumatetralyl with 55 ng/g). The results of the study indicate a serious toxicological risk for red foxes in Slovenia as part of the Western Balkans, and will contribute to the growing body of knowledge about the protection of European ecosystems, as wildlife is not limited by national borders.

Screening DNA Damage in the Rat Kidney and Liver by Untargeted DNA Adductomics.

Air pollution, tobacco smoke, and red meat are associated with renal cell cancer (RCC) risk in the United States and Western Europe; however, the chemicals that form DNA adducts and initiate RCC are mainly unknown. Aristolochia herbaceous plants are used for medicinal purposes in Asia and worldwide. They are a significant risk factor for upper tract urothelial carcinoma (UTUC) and RCC to a lesser extent. The aristolochic acid (AA) 8-methoxy-6-nitrophenanthro-[3,4-d]-1,3-dioxolo-5-carboxylic acid (AA-I), a component of Aristolochia herbs, contributes to UTUC in Asian cohorts and in Croatia, where AA-I exposure occurs from ingesting contaminated wheat flour. The DNA adduct of AA-I, 7-(2'-deoxyadenosin-N6-yl)-aristolactam I, is often detected in patients with UTUC, and its characteristic A:T-to-T:A mutational signature occurs in oncogenes and tumor suppressor genes in AA-associated UTUC. Identifying DNA adducts in the renal parenchyma and pelvis caused by other chemicals is crucial to gaining insights into unknown RCC and UTUC etiologies. We employed untargeted screening with wide-selected ion monitoring tandem mass spectrometry (wide-SIM/MS2) with nanoflow liquid chromatography/Orbitrap mass spectrometry to detect DNA adducts formed in rat kidneys and liver from a mixture of 13 environmental, tobacco, and dietary carcinogens that may contribute to RCC. Twenty DNA adducts were detected. DNA adducts of 3-nitrobenzanthrone (3-NBA), an atmospheric pollutant, and AA-I were the most abundant. The nitrophenanthrene moieties of 3-NBA and AA-I undergo reduction to their N-hydroxy intermediates to form 2'-deoxyguanosine (dG) and 2'-deoxyadenosine (dA) adducts. We also discovered a 2'-deoxycytidine AA-I adduct and dA and dG adducts of 10-methoxy-6-nitro-phenanthro-[3,4-d]-1,3-dioxolo-5-carboxylic acid (AA-III), an AA-I isomer and minor component of the herbal extract assayed, signifying AA-III is a potent kidney DNA-damaging agent. The roles of AA-III, other nitrophenanthrenes, and nitroarenes in renal DNA damage and human RCC warrant further study. Wide-SIM/MS2 is a powerful scanning technology in DNA adduct discovery and cancer etiology characterization.

Sex differences in 4-tert-octylphenol toxicokinetics: Exploration of sex as an effective covariate through an in vivo modeling approach.

4-tert-octylphenol (4-tert-OP) is a potentially harmful substance, which is found widely in the environment. Nevertheless, information on the in vivo toxicokinetics of 4-tert-OP is lacking, and quantitative risk assessment studies are urgently needed. Therefore, we aimed to quantitatively identify differences in the toxicokinetics of 4-tert-OP and its distribution among tissues between sexes. To this end, following exposure of male and female rats to 10 or 50 mg/kg 4-tert-OP orally and 4 or 8 mg/kg 4-tert-OP intravenously, we conducted a quantitative analysis of samples using ultra-high performance liquid chromatography-tandem mass spectrometry. The results revealed that the 4-tert-OP plasma concentration profiles differed between sexes; however, systemic absorption of 4-tert-OP through the gastrointestinal tract occurred within 0.5 h of exposure in both sexes. Although small, the excretion percentage of 4-tert-OP in urine and feces was lower in males than females (0.06-0.08% vs. 0.82-1.11% of exposure). Significant sex differences were also confirmed in the tissue distribution patterns of 4-tert-OP, and overall, the average tissue distribution in males was lower than that in females. The distribution of 4-tert-OP to liver, adipose, spleen, kidney, brain, and lung in both sexes was predominant. A covariate exploration modeling approach revealed that sex explained the differences in 4-tert-OP toxicokinetics between sexes. These significant differences in the toxicokinetics and tissue distribution of 4-tert-OP between sexes will be important for the scientific precision human risk assessment of 4-tert-OP.

Trace elements in the muscle and liver tissues of Garra shamal from the freshwater ecosystem of Oman: an exposure risk assessment.

Anthropogenic activities lead to environmental contamination with foreign substances such as heavy metals. This work was aimed to monitor trace elements (total arsenic (As), cadmium (Cd), chrome (Cr), cobalt (Co), copper (Cu), lead (Pb), manganese (Mn), mercury (Hg), nickel (Ni), and zinc (Zn)) contamination levels (dry weight base) in three natural freshwater reservoirs of Oman including Al Khawd and Al Amarat (Muscat Governorate) and Surur area (Ad Dakhiliyah Governorate as control area) using a native benthic inland fish (Garra shamal; Cyprinidae) for the first time. The muscle and liver of a hundred and twenty G. shamal were collected to assess the degree of metal contamination. Atomic absorption spectrometry was used as an analytical technique. From the spectrum of analyzed elements, we found Zn as a major element in monitored areas. The statistically significant (P < 0.05) highest concentrations of Zn liver (0.275 ± 0.065 µg/g) were in Al Amarat compared to the other areas. The concentrations of monitored elements in the fish muscle were lower than the liver samples. Furthermore, the fish length was significantly correlated with the accumulation of Hg and Co in both muscle and liver samples. In all analyzed fish from Oman inland water, the concentrations of elements were below the permissible limits; however, additional research is needed.

Full-length transcriptomics study of Ustiloxins-induced hepatocyte injury.

Ustiloxins is a mycotoxin produced by the metabolism of Rice false smut. Studies have shown that Ustiloxins may be toxic to animals, but there is still a lack of toxicological evidence. The liver, as the main organ for the biotransformation of foreign chemicals, may be the direct target organ of Ustiloxins toxicity. In this study, we found that cell viability decreased in a dose- and time-dependent manner when BNL CL.2 cells were treated with different concentrations of Ustiloxins (0, 5, 10, 20, 30, 40, 60, 80, 100, 150 and 200 µg/mL) for 24 and 48 h. In addition, scanning electron microscope observation showed that the cell membrane of the experimental group was damaged, with the appearance of apoptotic bodies. Moreover, the ROS and GSH levels were significantly increased in cells exposed to Ustiloxins. We analyzed the key action targets of Ustiloxins on hepatocyte injury using full-length transcriptomics. A total of 1099 differentially expressed genes were screened, of which 473 genes were up-regulated, and 626 genes were down-regulated. Besides, we also found that the expression of MCM7 and CDC45 in BNL CL.2 cells treated with Ustiloxins decreased, and the expression of CCl-2, CYP1b1, CYP4f13, and GSTM1 increased according to qRT-PCR. Ustiloxins might change CYP450 and GST-related genes, affect DNA replication and cell cycle, and lead to oxidative stress and liver cell injury.

Stereoselective bioaccumulation of chiral anticoagulant rodenticides in the liver of predatory and scavenging raptors.

Second-generation anticoagulant rodenticides (SGARs) are persistent chiral pesticides used to control rodent populations. Raptors are protected species and may be exposed through the ingestion of rodents contaminated with SGARs. Commercial formulations of SGARs are a mixture of four stereoisomers (E1, E2, E3, E4): the cis- and trans-diastereoisomers are each a racemic mixture of two enantiomers. In this study, the residue levels of all SGARs (bromadiolone, difenacoum, brodifacoum, difethialone, flocoumafen) were evaluated in the liver of 529 raptor carcasses. All species (n = 18) and 75 % of individuals (n = 396) were SGAR positive and 29 % (n = 154) had summed hepatic concentrations above 100 ng/g ww. Concentrations were higher for predators with facultative scavenging behaviors than for predators and obligate scavengers. Bromadiolone, brodifacoum and difenacoum had equivalent hepatic prevalence (between 48.9 and 49.9 %), and difethialone was detected less frequently (31.7 %). Concentrations and enantiomeric fractions of the four stereoisomers of all SGARs are described in to demonstrate the biological enantioselectivity of these chiral pesticides in the food chain. A difference was observed between the proportions of SGARs diastereoisomers and stereoisomers in the liver of all raptor species and in commercial baits. The enantioselective bioaccumulation of E1-trans-bromadiolone, E3-cis-brodifacoum, E1-cis-difenacoum and E3-cis-difethialone was characterized and represented 96.8 % of total SGARs hepatic residues. While hepatic concentrations were heterogeneous, the proportions of stereoisomers and diastereoisomers were homogeneous with no inter-individual or inter-species differences (only E1-trans-bromadiolone is present in hepatic residues). However, proportions of brodifacoum stereoisomers and diastereoisomers were more scattered, probably due to their slower elimination. This could provide an opportunity to date the exposure of individuals to brodifacoum. We highlight the need to consider each SGAR as four molecular entities (four stereoisomers) rather than one. These findings suggest new commercial formulations with the less persistent stereoisomers could reduce secondary exposure of non-target species.

Effect of urbanization on the trace element concentrations in the kidney, liver and spines of the European hedgehog (Erinaceus europaeus).

Urbanized environments may be suitable for some wild species, like the European hedgehog (Erinaceus europaeus). However, the (sub)urban areas are source of several pollutants, such as trace elements (TEs). The main objective of our study is to investigate the role of the European hedgehog as a potential bioindicator species for seven TEs (As, Cd, Cr, Cu, Ni, Pb, Zn) in a big urban agglomeration. We analyzed the kidney, liver and spines of 50 European hedgehogs. Moreover, we carried out geographic information system (GIS) with 31 individuals to examine the effects of human pressure (human density and urbanization) on TE concentrations in tissues. We detected the 7 TEs in all tissues. Trace elements, especially Pb and Zn, in the liver, kidney and spines seem to be influenced by human density and urbanization.