Three-dimensional graphene aerogel mitigated the toxic impact of chloramphenicol wastewater on microorganisms in an EGSB reactor.

Anaerobic treatment of chloramphenicol wastewater holds significant promise due to its potential for bioenergy generation. However, the high concentration of organic matter and residual toxic substances in the wastewater severely inhibit the activity of microorganisms. In this study, a three-dimensional graphene aerogel (GA), as a conductive material with high specific surface area (114.942 m2 g-1) and pore volume (0.352 cm3 g-1), was synthesized and its role in the efficiency and related mechanism for EGSB reactor to treat chloramphenicol wastewater was verified. The results indicated that synergy effects of GA for Chemical Oxygen Demand (COD) removal (increased by 8.17 %), chloramphenicol (CAP) removal (increased by 4.43 %) and methane production (increased by 70.29 %). Furthermore, GA increased the average particle size of anaerobic granular sludge (AGS) and promoted AGS to secrete more redox active substances. Microbial community analysis revealed that GA increased the relative abundance of functional bacteria and archaea, specifically Syntrophomonas, Geobacter, Methanothrix, and Methanolinea. These microbial species can participate in direct interspecific electron transfer (DIET). This research serves as a theoretical foundation for the application of GA in mitigating the toxic impact of refractory organic substances, such as antibiotics, on microorganisms during anaerobic treatment processes.

Simultaneous removal of norfloxacin and chloramphenicol using cold atmospheric plasma jet (CAPJ): Enhanced performance, synergistic effect, plasma-activated water (PAW) contribution, mechanism and toxicity evaluation.

The extensive abuse and inadvertent discharge of various antibiotics into the environment has become a serious problem for posing a big threat to human health. In order to deal with this problem, we utilized cold atmospheric plasma jet (CAPJ) to treat two different antibiotics, namely, norfloxacin and chloramphenicol, and investigated the efficiencies and corresponding mechanisms for removing the mixed antibiotics. In the application of the CAPJ technique, we made use of not only the direct plasma processing, but also the indirect plasma-activated water (PAW) treatment. The efficiency for mixed antibiotics treatment was considerably enhanced as compared to the efficiency for treatment of single antibiotics. The contributions from the CAPJ-induced reactive oxygen/nitrogen species (RONS) were examined, showing that ·OH and 1O2 played a major role in the degradation of norfloxacin and chloramphenicol in the direct plasma treatment, while 1O2 played a major role in the PAW treatment. The bio-toxicity evaluation was also provided to verify the ecological safety of the CAPJ treatment. As such, this work has not only showed the effectiveness of CAPJ treatment of mixed antibiotics, but also elucidated the mechanisms for the enhanced treatment efficiency, which may provide a new solution for treatment of antibiotics in the environment.

Higher toxicity induced by co-exposure of polystyrene microplastics and chloramphenicol to Microcystis aeruginosa: Experimental study and molecular dynamics simulation.

Antibiotics and microplastics (MPs) inevitably coexist in natural waters, but their combined effect on aquatic organisms is still ambiguous. This study investigated the individual and combined toxicity of chloramphenicol (CAP) and micro-polystyrene (mPS) particles to Microcystis aeruginosa by physiological biomarkers, related gene expression, and molecular dynamics simulation. The results indicated that both individual and joint treatments threatened algal growth, while combined toxicity was higher than the former. Photosynthetic pigments and gene expression were inhibited by single CAP and mPS exposure, but CAP dominated and aggravated photosynthetic toxicity in combined exposure. Additionally, mPS damaged cell membranes and induced oxidative stress, which might further facilitate the entry of CAP into cells during co-exposure. The synergistic effect of CAP and mPS might be explained by the common photosynthetic toxicity target of CAP and mPS as well as oxidative stress. Furthermore, the molecular dynamics simulation revealed that CAP altered conformations of photosynthetic assembly protein YCF48 and SOD enzyme, and competed for functional sites of SOD, thus disturbing photosynthesis and antioxidant systems. These findings provide useful insights into the combined toxicity mechanism of antibiotics and MPs as well as highlight the importance of co-pollutant toxicity in the aquatic environment.

Metal-containing landfills as a source of antibiotic tolerance.

To unveil the potential effect of metal presence to antibiotic tolerance proliferation, four sites of surface landfills containing tailings from metal processing in Slovakia (Hnústa, Hodrusa, Kosice) and Poland (Tarnowskie Góry) were investigated. Tolerance and multitolerance to selected metals (Cu, Ni, Pb, Fe, Zn, Cd) and antibiotics (ampicillin, tetracycline, chloramphenicol, and kanamycin) and interrelationships between them were evaluated. A low bacterial diversity (Shannon-Wiener index from 0.83 to 2.263) was detected in all sampling sites. Gram-positive bacteria, mostly belonging to the phylum Actinobacteria, dominated in three of the four sampling sites. The recorded percentages of tolerant bacterial isolates varied considerably for antibiotics and metals from 0 to 57% and 0.8 to 47%, respectively, among the sampling sites. Tolerances to chloramphenicol (45-57%) and kanamycin (32-45%) were found in three sites. Multitolerance to several metals and antibiotics in the range of 24 to 48% was recorded for three sites. A significant positive correlation (p < 0.05) for the co-occurrence of tolerance to each studied metal and at least one of the antibiotics was observed. Exposure time to the metal (landfill duration) was an important factor for the development of metal- as well as antibiotic-tolerant isolates. The results show that metal-contaminated sites represent a significant threat for human health not only for their toxic effects but also for their pressure to antibiotic tolerance spread in the environment.

Effects of natural nanoparticles on the acute toxicity, chronic effect, and oxidative stress response of phenicol antibiotics in Daphnia magna.

Natural nanoparticles (NNP) are ubiquitous in natural water and can interact with other contaminants, causing ecotoxic effects on aquatic nontarget organisms. However, the impact of NNPs on the ecotoxicity of antibiotics remains largely unknown. This work investigated the acute toxicity, chronic effect, and oxidative response and damage in Daphnia magna co-exposed to phenicol antibiotics (chloramphenicol, thiamphenicol) and different concentrations of NNPs (10 mg/L: environmentally relevant concentration; 100 mg/L: a high concentration that caused no apparent immobilization in D. magna). The results showed that the acute toxicity of chloramphenicol was increased by 10 mg/L NNPs but decreased by 100 mg/L NNPs; both concentrations of NNPs increased and decreased acute toxicities of thiamphenicol and chloramphenicol + thiamphenicol treatments, respectively. After long-term exposure, phenicol antibiotics (1 µg/L) and NNP (10 mg/L) mixtures in environmentally relevant concentrations significantly affected the reproduction of D. magna but did not influence their growth. The catalase activity, reduced glutathione level, and malonaldehyde content in D. magna also varied with the NNPs concentrations. Notably, the lowest concentration of thiamphenicol and chloramphenicol + thiamphenicol combined with NNPs significantly increased the malondialdehyde content in D. magna compared with the control, indicating membrane lipid peroxidation occurred in daphnids. This study suggests that the toxic effects of contaminants and NNPs on aquatic organisms should be considered thoroughly to avoid underestimating the hazard of these pollutants in the actual aquatic environment.

Removal of chloramphenicol antibiotics in natural and engineered water systems: Review of reaction mechanisms and product toxicity.

Chloramphenicol antibiotics are widely applied in human and veterinary medicine. They experience natural attenuation and/or chemical degradation during oxidative water treatment. However, the environmental risks posed by the transformation products of such organic contaminants remain largely unknown from the literature. As such, this review aims to summarize and analyze the elimination efficiency, reaction mechanisms, and resulting product risks of three typical chloramphenicol antibiotics (chloramphenicol, thiamphenicol, and florfenicol) from these transformation processes. The obtained results suggest that limited attenuation of these micropollutants is observed during hydrolysis, biodegradation, and photolysis. Comparatively, prominent abatement of these compounds is accomplished using advanced oxidation processes; however, efficient mineralization is still difficult given the formation of recalcitrant products. The in silico prediction on the multi-endpoint toxicity and biodegradability of different products is systematically performed. Most of the transformation products are estimated with acute and chronic aquatic toxicity, genotoxicity, and developmental toxicity. Furthermore, the overall reaction mechanisms of these contaminants induced by multiple oxidizing species are revealed. Overall, this review unveils the non-overlooked and serious secondary risks and biodegradability recalcitrance of the degradation products of chloramphenicol antibiotics using a combined experimental and theoretical method. Strategic improvements of current treatment technologies are strongly recommended for complete water decontamination.

Effect of chloramphenicol on the life table demography of Brachionus calyciflorus (Rotifera): A multigenerational study.

During the last two decades, there has been increasing concerns about the presence of antibiotics in aquatic environments. Phenicol antibiotics, such as chloramphenicol (CMP), commonly used in the veterinary and aquaculture fields to treat infections, have been often detected in aquatic environments, but scarce ecotoxicity information regarding the effects of CMP on non-target aquatic organisms is available, and multigenerational studies are seldom studied. Here we quantified the demographic responses of Brachionus calyciflorus exposed to sublethal concentrations (0, 5, 10, 30, 50, 70 and 90 mg L-1) of CMP for three successive generations (P0, F1, and F2). Our results showed that compared to the control, higher concentrations of CMP significantly decreased the life expectancy at hatching, generation time, net reproductive rate and intrinsic rate of population increase in all three generations, and the proportion of mictic offspring in the F1 generation of B. calyciflorus. With increasing generations, higher concentrations of CMP showed increased toxic effects on life expectancy at hatching and net reproductive rate, but irregular negative effects on generation time, intrinsic rate of population increase, and proportion of mictic offspring of the rotifers. These results indicate that multigenerational studies are necessary to prevent insufficient assessments of the impact of antibiotics in aquatic ecosystems.

Probabilistic Risk Assessment of Dietary Exposure to Chloramphenicol in Guangzhou, China.

Chloramphenicol has been used in veterinary medicine, where its residues can remain in food of animal origin, thus potentially causing adverse health effects. This facilitated the ban for its use in food-producing animals globally, but its residues have remained ubiquitous. In this study, food commodities possibly contaminated with chloramphenicol, including livestock meat, poultry, edible viscera, fish, shrimp and crab, molluscs, milk, and eggs, were collected from domestic retail shops in all the 11 districts of Guangzhou and tested for its residue. Probabilistic risk assessment model calculations for its dietary exposure, and the margin of exposure (displayed as mean values and 5th percentile to 95th percentile ranges) were performed by using @RISK software based on a Monte Carlo simulation with 10,000 iterations. The results indicated the detection of chloramphenicol in 248 out of 1454 samples (17.1%), which averaged to a level of 29.1 µg/kg. The highest average value was observed in molluscs (148.2 µg/kg, with the top value as 8196 µg/kg); meanwhile, based on the dietary structure of a typical Cantonese, pond fish, pork, and poultry meat contributed most (about 80%) to the residents' dietary exposure to chloramphenicol. The margin of exposure for dietary chloramphenicol exposure in Guangzhou residents was 2489, which was apparently below 5000 (the borderline for judging a health risk), particularly low in preschool children (2094, suggesting an increased risk). In conclusion, the study suggests that chloramphenicol exposure in Guangzhou residents is considerable, and its relevant health hazard, especially for preschool children, is worthy of further investigation.

The Influence of Metabolic Inhibitors, Antibiotics, and Microgravity on Intact Cell MALDI-TOF Mass Spectra of the Cyanobacterium Synechococcus Sp. UPOC S4.

The aim and novelty of this paper are found in assessing the influence of inhibitors and antibiotics on intact cell MALDI-TOF mass spectra of the cyanobacterium Synechococcus sp. UPOC S4 and to check the impact on reliability of identification. Defining the limits of this method is important for its use in biology and applied science. The compounds included inhibitors of respiration, glycolysis, citrate cycle, and proteosynthesis. They were used at 1-10 µM concentrations and different periods of up to 3 weeks. Cells were also grown without inhibitors in a microgravity because of expected strong effects. Mass spectra were evaluated using controls and interpreted in terms of differential peaks and their assignment to protein sequences by mass. Antibiotics, azide, and bromopyruvate had the greatest impact. The spectral patterns were markedly altered after a prolonged incubation at higher concentrations, which precluded identification in the database of reference spectra. The incubation in microgravity showed a similar effect. These differences were evident in dendrograms constructed from the spectral data. Enzyme inhibitors affected the spectra to a smaller extent. This study shows that only a long-term presence of antibiotics and strong metabolic inhibitors in the medium at 10-5 M concentrations hinders the correct identification of cyanobacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF).

How to disinfect anuran eggs? Sensitivity of anuran embryos to chemicals widely used for the disinfection of larval and post-metamorphic amphibians.

Emerging infectious diseases are major drivers of global and local amphibian biodiversity loss. Therefore, developing effective disinfection methods to manage the impact of diseases in wild and captive "ark" populations are an important goal in amphibian conservation. While chemical disinfectants have been used safely and effectively in larval and adult amphibians infected with pathogenic microbes, their applicability to amphibian egg masses has remained untested. To bridge this gap, we exposed embryos of the common toad (Bufo bufo) and agile frog (Rana dalmatina) experimentally to three widely used disinfectants: voriconazole, chloramphenicol and chlorogen-sesquihydrate. For 3 days we exposed portions of egg masses to these disinfectants at 1×, 2×, 5× and 10× the concentration recommended for the disinfection of tadpoles and adults. Subsequently, we recorded embryonic and larval survival, as well as larval body mass and the incidence of abnormalities 12 days after hatching. Application of voriconazole had species- and concentration-dependent negative impacts on survival and body mass, and caused marked malformations in the viscerocranial structure of B. bufo tadpoles. Exposure to chlorogen-sesquihydrate also resulted in significant mortality in B. bufo embryos and negatively affected body mass of R. dalmatina larvae. Chloramphenicol had little negative effects on embryos or larvae in either species. Based on these results, the application of voriconazole and chlorogen-sesquihydrate cannot be recommended for the disinfection of amphibian eggs, whereas treatment with chloramphenicol appears to be a safe method for eliminating potential pathogens from anuran egg masses and their immediate aquatic environment.

Mixture toxicity effects of chloramphenicol, thiamphenicol, florfenicol in Daphnia magna under different temperatures.

Acute toxicities of chloramphenicol (CAP), thiamphenicol (TAP), and florfenicol (FLO) and their mixtures on Daphnia magna under two representative temperatures of the aquatic environment (20 and 25 °C) have been examined. Their toxicities depicted with an order of 72-h EC50 values were as follows: CAP > FLO > TAP and CAP ≈ FLO > TAP under 20 and 25 °C, separately. Furthermore, the acute toxicity significantly increased with the rise of temperature from 20 to 25 °C in nearly all separate and mixture phenicol antibiotics. Meanwhile, the most toxic combination under two different temperatures was diverse. The nature of toxicological interactions of phenicol antibiotic mixtures was analyzed by Combination Index (CI) equation. In general, a dual synergism-antagonism effect was dominant in nearly all mixtures at both temperatures. The prediction suitability of Concentration Addition (CA), Independent Action (IA) models, and CI method was compared, suggesting that the CI equation seems to be more appropriate for predicting the toxicity values of phenicol drugs than CA and IA models. In brief, phenicol antibiotic mixtures with temperature variation may pose more significant hazards and risks to aquatic organisms; hence, the environment.

Growth and Cellular Responses of Toxigenic Microcystis to Chloramphenicol-Stress at Various Environmentally-Relevant Nitrogen Levels.

This study explored nitrogen (N)-dependent interaction between Microcystis and chloramphenicol (CAP) along 20 day-test. Results showed that 5 mg/L N largely alleviated inhibitory effects of CAP on Microcystis growth, while 50 and 0.5 mg/L N exacerbated growth-inhibition by CAP especially in early (before day 8) and mid-late stage, respectively. At each N level, CAP-induced antioxidant defense and cell damage extents were negatively correlated to growth state in each stage, and CAP-biodegradation coincided with Microcystis growth and glutathione synthesis dynamics, implying that antioxidant defense, cell damage and CAP-removal closely linked to N-dependent Microcystis growth under CAP-stress. Microcystin (MC)-production and -release under CAP-stress were also N-dependent. Although Microcystis growth was greatly-inhibited by prolonged CAP-stress at 0.5 mg/L N, delayed CAP-loss and high MC-release at 0.5 mg/L N should be emphasized during Microcystis-dominated cyanobacterial blooms (MCBs) and CAP co-occurrence. This study had great implication in risk assessment for MCBs-CAP co-occurrence in different waters.

Comparison of the Anti-Inflammatory and Cytotoxic Potential of Different Corticosteroid Eye Drop Preparations.

PURPOSE: To compare the immunosuppressive and cytotoxic effects of three anti-inflammatory eye drops formulations containing betamethasone plus chloramphenicol (B+C), dexamethasone plus netilmicin (D+N) or dexamethasone plus tobramycin (D+T).Methods: The eye drops formulations have been tested at different dilutions on cytokine synthesis by mouse or human cultured macrophages, as well as proliferation and viability of cultured human corneal cells (HCE).Results: B+C reduced IL6 and TNFα production by cultured mouse or human macrophages more potently than D+N and D+T, with the tree formulations having the same impact on IL-10 expression. We also found that the eye drops preparations reduced proliferation of HCE cells, with D+T showing the higher anti-proliferative potency and B+C showing the lower cytotoxic potential.Conclusion: Our study points out that it may be erroneous to consider routinely-used anti-inflammatory eye drops preparations with analogous formulations as readily interchangeable and of similar potency and tolerability.

Phosphorus Influences the Interaction Between Toxigenic Microcystis and Chloramphenicol.

Microcystis growth and physiological responses to chloramphenicol (CAP)-stress were explored at different phosphorus (P) concentrations during 20-day exposure. Under CAP-stress, Microcystis exhibited (i) stronger total protein synthesis and antioxidant defenses at 5 mg/L P than 0.05-0.5 mg/L P in early test period (before day 8), and (ii) greater CAP-removal via biodegradation at 5 mg/L P in mid-late period. Due to above mechanisms, 5 mg/L P largely alleviated the inhibitory effect of CAP on Microcystis growth until test end, thus minimizing CAP toxicity to Microcystis, compared with 0.05-0.5 mg/L P. Moreover, microcystin-production and -release by Microcystis under CAP-stress were also P-dependent. These results suggested that under CAP-stress, although Microcystis growth was more inhibited at 0.05-0.5 mg/L P, higher microcystin-release and CAP residual at 0.05-0.5 mg/L P than at 5 mg/L P still caused eco-risks, which had important implication for risk assessment during Microcystis-dominated blooms and CAP pollution co-occurrence in different waters.

Evaluation of the subtle effects and oxidative stress response of chloramphenicol, thiamphenicol, and florfenicol in Daphnia magna.

Phenicol antibiotics, such as chloramphenicol, thiamphenicol, and florfenicol, are commonly used in the veterinary and aquaculture fields to treat diseases and have frequently been detected in aquatic environments. Nevertheless, there is limited information regarding the effects of phenicol antibiotics on aquatic nontarget species. Thus, the present study aims to investigate the long-term (21-d) influence on the reproduction and growth of and the acute (24-h) oxidative response and tissue damage in the crustacean Daphnia magna after exposure to phenicol drugs, including their environmental concentrations. The results indicate that D. magna exposed to florfenicol are likely to cause more adverse effects than those exposed to chloramphenicol and thiamphenicol over long-term (21-d) exposures. Furthermore, changes in biochemical biomarkers such as malondialdehyde (MDA), catalase (CAT), and reduced glutathione (GSH) induced by individual and mixtures of phenicol antibiotics were also observed. Low concentrations of chloramphenicol, thiamphenicol + florfenicol, and chloramphenicol + thiamphenicol significantly increased the MDA levels of D. magna after 24-h exposures, causing cellular oxidative damage in the animals. In addition, discrepancies between CAT activities and GSH levels were observed, underscoring the need to evaluate multiple indicators of oxidative stress in toxicological studies using D. magna as a model. Environ Toxicol Chem 2019;38:575-584. © 2018 SETAC.

New insight into the toxic effects of chloramphenicol and roxithromycin to algae using FTIR spectroscopy.

Antibiotics have been frequently detected in the aquatic environment, and they may affect aquatic organisms such as algae. Here we investigated toxicity of chloramphenicol (CAP) and roxithromycin (ROX) on four species of green algae (Pseudokirchneriella subcapitata, Scenedesmus quadricauda, Scenedesmus obliquus, and Scenedesmus acuminatus) at biochemical level by Fourier transform infrared spectroscopy (FTIR). The results revealed that both CAP and ROX had negative effects on algal growth and caused alterations of biochemical components. The toxic effects varied among the four algal species and S. acuminatus was found to be less sensitive than the other three species to the antibiotics. Even with similar mechanism of action, ROX displayed more adverse effects to algae than CAP. Both antibiotics could affect algae by inhibiting fatty acid synthesis and promoting protein and DNA aggregation, thus leading to accumulation of lipid peroxidation products, increment of the loose ß-sheet structure protein and transformation of B-DNA to Z-DNA. The findings from this study revealed the toxic mechanism of antibiotics to algae at the biochemical level.

Evaluation of drug-induced hematotoxicity using novel in vitro monkey CFU-GM and BFU-E colony assays.

In order to evaluate drug-induced hematotoxicity in monkey cells in vitro, colony-forming unit-granulocyte, macrophage (CFU-GM), and burst-forming unit-erythroid (BFU-E) colony assays were established using mononuclear cells in the bone marrow collected from male cynomolgus monkeys. Furthermore, the effects of doxorubicin, chloramphenicol, and linezolid on CFU-GM and BFU-E colony formation were investigated using established monkey CFU-GM and BFU-E colony assays in comparison with those on human CFU-GM and BFU-E colonies acquired from human umbilical cord blood cells. Bone marrow mononuclear cells were collected from the ischial or iliac bone of male cynomolgus monkeys. The cells were subsequently processed by density gradient separation at 1.067, 1.070, or 1.077 g/mL for CFU-GM or 1.077 g/mL for BFU-E, and then cultured in methylcellulose medium for 9 or 13 days, respectively. A sufficient number of CFU-GM colonies were formed from mononuclear cells processed at a density of 1.070 g/mL. Moreover, the number of BFU-E colonies from the cells processed at a density of 1.077 g/mL was sufficient for the colony assay. The number of CFU-GM or BFU-E colonies decreased after treatment with the drugs of interest in a concentration-dependent manner. Compared with human CFU-GM, monkey CFU-GM were more sensitive to chloramphenicol and resistant to doxorubicin, whereas monkey BFU-E were more sensitive to all compounds in comparison to the sensitivity of human BFU-E. In conclusion, monkey CFU-GM and BFU-E colony assays were established and considered useful tools to evaluate the differences in drug-induced hematotoxicity between species.

Determination of Aflatoxin M1 and Chloramphenicol in Milk Based on Background Fluorescence Quenching Immunochromatographic Assay.

Harsh demanding has been exposed on the concentration of aflatoxin M1 (AFM1) and chloramphenicol (CAP) in milk. In this study, we developed a new method based on background fluorescence quenching immunochromatographic assay (bFQICA) to detect AFM1 and CAP in milk. The detection limit for AFM1 was 0.0009 ng/mL, while that for the CAP was 0.0008 ng/mL. The assay variability was determined with 3 AFM1 standards (i.e., 0.25 ng/mL, 0.5 ng/mL, and 1.0 ng/mL), and the actual detection value was 0.2497, 0.5329, and 1.0941, respectively. For the assay variability of 3 CAP standards (i.e., 0.10 ng/mL, 0.30 ng/mL, and 0.50 ng/mL), the actual detection value was 0.0996, 0.3096, and 0.4905, respectively. The recovery rate of AFM1 was 99.7%-101.7%, while that for CAP was 95.3%-97.6%. For the test stability, AFM1 and CAP showed satisfactory test stability even at month 5. Compared with the sensitivity of liquid chromatography-mass spectrometry (LC-MS) method, no statistical difference was noticed in results of the bFQICA. Our method is convenient for the detection of AFM1 and CAP in milk with a test duration of about 8 minutes. Additionally, an internal WiFi facility is provided in the system allowing for quick connection and storage in the intelligent cell phone.

Occurrence of chloramphenicol in cereal straw in north-western Europe.

Two surveys are presented of straw analysed for naturally occurring chloramphenicol (CAP), a drug banned for use in food-producing animals. In the first study, CAP was analysed by LC-MS/MS and detected in 37 out of 105 straw samples originating from the Netherlands, France, the UK, Germany and Denmark. The highest level found was 6.3 µg kg(-1), the average 0.6 µg kg(-1) and the median 0.2 µg kg(-1). The second study included a method comparison between ELISA and LC-MS/MS and a survey of CAP in cereal straw sampled at farms in all areas of Sweden. A total of 215 samples were screened by ELISA and a subset of 26 samples was also analysed by LC-MS/MS. Fifty-four of the samples contained more than 1 µg kg(-1) CAP and the highest level found was 32 µg kg(-1) (confirmed by LC-MS/MS). The highest contents of CAP in this study were allocated to the Baltic sea coast in the south-eastern part of Sweden (the county of Skåne and the Baltic Sea isle of Gotland). These results indicate a high incidence of CAP in straw in north-west Europe and have a severe impact on the enforcement of European Union legislation.

Classification of Cholestatic and Necrotic Hepatotoxicants Using Transcriptomics on Human Precision-Cut Liver Slices.

Human toxicity screening is an important stage in the development of safe drug candidates. Hepatotoxicity is one of the major reasons for the withdrawal of drugs from the market because the liver is the major organ involved in drug metabolism, and it can generate toxic metabolites. There is a need to screen molecules for drug-induced hepatotoxicity in humans at an earlier stage. Transcriptomics is a technique widely used to screen molecules for toxicity and to unravel toxicity mechanisms. To date, the majority of such studies were performed using animals or animal cells, with concomitant difficulty in interpretation due to species differences, or in human hepatoma cell lines or cultured hepatocytes, suffering from the lack of physiological expression of enzymes and transporters and lack of nonparenchymal cells. The aim of this study was to classify known hepatotoxicants on their phenotype of toxicity in humans using gene expression profiles ex vivo in human precision-cut liver slices (PCLS). Hepatotoxicants known to induce either necrosis (n = 5) or cholestasis (n = 5) were used at concentrations inducing low (<30%) and medium (30-50%) cytotoxicity, based on ATP content. Random forest and support vector machine algorithms were used to classify hepatotoxicants using a leave-one-compound-out cross-validation method. Optimized biomarker sets were compared to derive a consensus list of markers. Classification correctly predicted the toxicity phenotype with an accuracy of 70-80%. The classification is slightly better for the low than for the medium cytotoxicity. The consensus list of markers includes endoplasmic reticulum stress genes, such as C2ORF30, DNAJB9, DNAJC12, SRP72, TMED7, and UBA5, and a sodium/bile acid cotransporter (SLC10A7). This study shows that human PCLS are a useful model to predict the phenotype of drug-induced hepatotoxicity. Additional compounds should be included to confirm the consensus list of markers, which could then be used to develop a biomarker PCR-array for hepatotoxicity screening.