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  1. Hadibarata T, Kristanti RA
    Bioresour Technol, 2012 Mar;107:314-8.
    PMID: 22209445 DOI: 10.1016/j.biortech.2011.12.046
    Armillaria sp. F022, a white-rot fungus isolated from a tropical rain forest in Samarinda, Indonesia, was used to biodegrade benzo[a]pyrene (BaP). Transformation of BaP, a 5-ring polycyclic aromatic hydrocarbon (PAH), by Armillaria sp. F022, which uses BaP as a source of carbon and energy, was investigated. However, biodegradation of BaP has been limited because of its bioavailability and toxicity. Five cosubstrates were selected as cometabolic carbon and energy sources. The results showed that Armillaria sp. F022 used BaP with and without cosubstrates. A 2.5-fold increase in degradation efficiency was achieved after addition of glucose. Meanwhile, the use of glucose as a cosubstrate could significantly stimulate laccase production compared with other cosubstrates and not using any cosubstrate. The metabolic pathway was elucidated by identifying metabolites, conducting biotransformation studies, and monitoring enzyme activities in cell-free extracts. The degradation mechanism was determined through the identification of several metabolites: benzo[a]pyrene-1,6-quinone, 1-hydroxy-2-benzoic acid, and benzoic acid.
  2. Aziz A, Agamuthu P, Alaribe FO, Fauziah SH
    Environ Technol, 2018 Feb;39(4):527-535.
    PMID: 28281885 DOI: 10.1080/09593330.2017.1305455
    Benzo[a]pyrene is a high-molecular-weight polycyclic aromatic hydrocarbon highly recalcitrant in nature and thus harms the ecosystem and/or human health. Therefore, its removal from the marine environment is crucial. This research focuses on benzo[a]pyrene degradation by using enriched bacterial isolates in consortium under saline conditions. Bacterial isolates capable of using benzo[a]pyrene as sole source of carbon and energy were isolated from enriched mangrove sediment. These isolates were identified as Ochrobactrum anthropi, Stenotrophomonas acidaminiphila, and Aeromonas salmonicida ss salmonicida. Isolated O. anthropi and S. acidaminiphila degraded 26% and 20%, respectively, of an initial benzo[a]pyrene concentration of 20 mg/L after 8 days of incubation in seawater (28 ppm of NaCl). Meanwhile, the bacterial consortium decomposed 41% of an initial 50 mg/L benzo[a]pyrene concentration after 8 days of incubation in seawater (28 ppm of NaCl). The degradation efficiency of benzo[a]pyrene increased to 54%, when phenanthrene was supplemented as a co-metabolic substrate. The order of biodegradation rate by temperature was 30°C > 25°C > 35°C. Our results suggest that co-metabolism by the consortium could be a promising biodegradation strategy for benzo[a]pyrene in seawater.
  3. Karami A, Christianus A, Ishak Z, Shamsuddin ZH, Masoumian M, Courtenay SC
    J Hazard Mater, 2012 May 15;215-216:108-14.
    PMID: 22417397 DOI: 10.1016/j.jhazmat.2012.02.038
    This study examined the potential of Pseudomonas aeruginosa abundance in the intestines of fish as an indicator of exposure to benzo[a]pyrene (BaP). P. aeruginosa populations were enumerated in juvenile African catfish (Clarias gariepinus) injected intramuscularly three days previous with 0, 10, 30, 40, 50 or 70mg/kg of BaP. Hepatic EROD and GST activities and biliary fluorescent aromatic compounds (FACs) 1-OH BaP, 3-OH BaP, 7,8-D BaP and BaP were quantified to investigate agreements between the new indicator and established fish biomarkers. The shape of bacterial population (logarithm of colony-forming unit) dose-response curve generally matched those of biliary FACs concentrations. Conversely, the EROD and GST dose-response curves were generally the mirror images of the bacterial population curve. Changes in intestinal P. aeruginosa population appear to be an indirect effect of BaP exposure because exposure to 0-100μg/ml BaP had no effect on P. aeruginosa populations grown on agar plates containing BaP. Using intestinal P. aeruginosa population of fish as a universal indicator of BaP pollution in aquatic environments is discussed.Conversely, the EROD and GST dose-response curves were generally the mirror images of the bacterial population curve.
  4. Karami A, Christianus A, Bahraminejad B, Gagné F, Courtenay SC
    Ecotoxicol Environ Saf, 2012 Mar;77:28-34.
    PMID: 22101109 DOI: 10.1016/j.ecoenv.2011.10.026
    This study examined the potential of artificial neural network (ANN) modeling to infer timing, route and dose of contaminant exposure from biomarkers in a freshwater fish. Hepatic glutathione S-transferase (GST) activity and biliary concentrations of BaP, 1-OH BaP, 3-OH BaP and 7,8D BaP were quantified in juvenile Clarias gariepinus injected intramuscularly or intraperitoneally with 10-50 mg/kg benzo[a]pyrene (BaP) 1-3 d earlier. A feedforward multilayer perceptron (MLP) ANN resulted in more accurate prediction of timing, route and exposure dose than a linear neural network or a radial basis function (RBF) ANN. MLP sensitivity analyses revealed contribution of all five biomarkers to predicting route of exposure but no contribution of hepatic GST activity or one of the two hydroxylated BaP metabolites to predicting time of exposure and dose of exposure. We conclude that information content of biomarkers collected from fish can be extended by judicious use of ANNs.
  5. Agus BAP, Hussain N, Selamat J
    Food Chem, 2020 Jan 15;303:125398.
    PMID: 31470272 DOI: 10.1016/j.foodchem.2019.125398
    Roasting is an important process in cocoa production which may lead to formation of non-desirable compounds such as polycyclic aromatic hydrocarbons (PAHs). Therefore, PAH4 (sum of four different polycyclic aromatic hydrocarbons; benz[a]anthracene, chrysene, benzo[b]fluoranthene, and benzo[a]pyrene) in roasted cocoa beans was determined using a modified method (combination of QuEChERS and DLLME), and quantified by HPLC-FLD. The modified method was validated and met the performance criteria required by the EU Regulation (No. 836/2011). Results show a significant (p 
  6. Singh A, Banerjee T, Latif MT, Ramanathan S, Suradi H, Othman M, et al.
    Chemosphere, 2023 Nov;340:139943.
    PMID: 37625487 DOI: 10.1016/j.chemosphere.2023.139943
    Abundance of fine particulate-bound 16 priority polycyclic aromatic hydrocarbons (PAHs) was investigated to ascertain its sources and potential carcinogenic health risks in Varanasi, India. The city represents a typical urban settlement of South Asia having particulate exposure manyfold higher than standard with reports of pollution induced mortalities and morbidities. Fine particulates (PM2.5) were monitored from October 2019 to May 2020, with 32% of monitoring days accounting ≥100 μgm-3 of PM2.5 concentration, frequently from November to January (99% of monitoring days). The concentration of 16 priority PAHs varied from 24.1 to 44.6 ngm-3 (mean: 33.1 ± 3.2 ngm-3) without much seasonal deviations. Both low (LMW, 56%) and high molecular weight (HMW, 44%) PAHs were abundant, with Fluoranthene (3.9 ± 0.4ngm-3) and Fluorene (3.5 ± 0.3ngm-3) emerged as most dominating PAHs. Concentration of Benzo(a)pyrene (B(a)P, 0.5 ± 0.1ngm-3) was lower than the national standard as it contributed 13% of total PAHs mass. Diagnostic ratios of PAH isomers indicate predominance of pyrogenic sources including emissions from biomass burning, and both from diesel and petrol-driven vehicles. Source apportionment using receptor model revealed similar observation of major PAHs contribution from biomass burning and fuel combustion (54% of source contribution) followed by coal combustion for residential heating and cooking purposes (44%). Potential toxicity of B[a]P equivalence ranged from 0.003 to 1.365 with cumulative toxicity of 2.13ngm-3. Among the PAH species, dibenzo[h]anthracene contributed maximum toxicity followed by B[a]P, together accounting 86% of PAH induced carcinogenicity. Incremental risk of developing cancer through lifetime exposure (ILCR) of PAHs was higher in children (3.3 × 10-4) with 56% contribution from LMW PAHs, primarily through ingestion and dermal contact. Adults in contrast, were more exposed to inhale airborne PAHs with cumulative ILCR of 2.2 × 10-4. However, ILCR to PM2.5 exposure is probably underestimated considering unaccounted metal abundance thus, require source-specific control measures.
  7. Hadibarata T, Kristanti RA
    J Environ Manage, 2012 Nov 30;111:115-9.
    PMID: 22835655 DOI: 10.1016/j.jenvman.2012.06.044
    The biodegradation of benzo[a]pyrene (BaP) by using Polyporus sp. S133, a white-rot fungus isolated from oil-contaminated soil was investigated. Approximately 73% of the initial concentration of BaP was degraded within 30 d of incubation. The isolation and characterization of 3 metabolites by thin layer chromatography, column chromatography, and UV-vis spectrophotometry in combination with gas chromatography-mass spectrometry, indicated that Polyporus sp. S133 transformed BaP to BaP-1,6-quinone. This quinone was further degraded in 2 ways. First, BaP-1,6-quinone was decarboxylated and oxidized to form coumarin, which was then hydroxylated to hydroxycoumarin, and finally to hydroxyphenyl acetic acid by addition of an epoxide group. Second, Polyporus sp. S133 converted BaP-1,6-quinone into a major product, 1-hydroxy-2-naphthoic acid. During degradation, free extracellular laccase was detected with reduced activity of lignin peroxidase, manganese-dependent peroxidase and 2,3-dioxygenase, suggesting that laccase and 1,2-dioxygenase might play an important role in the transformation of PAHs compounds.
  8. Latif IK, Karim AJ, Zuki AB, Zamri-Saad M, Niu JP, Noordin MM
    Histol Histopathol, 2011 06;26(6):699-710.
    PMID: 21472685 DOI: 10.14670/HH-26.699
    The immune response of broiler chickens exposed to intra-tracheal (i.t.) administration of benzo[a]pyrene (BaP) with and without Nigella sativa (Ns) supplementation was investigated. A total of 120 day-old chicks were divided into four groups comprising 30 birds each, into a control, Ns, BaP, and BaP+Ns group. Immune responses to Newcastle disease (ND) were evaluated by haemagglutination inhibition (HI), phytohaemagglutinin (PHA) skin test and carbon clearance assay (CCA). In most instances, there was a significant increase (p<0.05) in the ND-HI antibody titers, PHA skin-swelling response and phagocytic activity in the BaP + Ns group compared to that of the BaP group. Likewise, organ weight and indices of the spleen, bursa of Fabricius and thymus of birds from the BaP + Ns group were also higher (p<0.05) than that of the BaP group from day 1 until day 21. It is concluded that exposure to BaP may exert adverse effects on the immune system of broilers which may increase their susceptibility to disease, and Ns supplementation significantly reduces these alterations.
  9. Keshavarzifard M, Zakaria MP, Sharifi R
    Arch Environ Contam Toxicol, 2017 Oct;73(3):474-487.
    PMID: 28497299 DOI: 10.1007/s00244-017-0410-0
    The distribution, sources, and human health risk assessment of polycyclic aromatic hydrocarbons (PAHs) in surface sediment and the edible tissue of short-neck clam (Paphia undulata) from mudflat ecosystem in the west coast of Malaysia were investigated. The concentrations of ∑16 PAHs varied from 347.05 to 6207.5 and 179.32 to 1657.5 ng g-1 in sediment and short-neck clam samples, respectively. The calculations of mean PEL quotients (mean-PELQs) showed that the ecological risk of PAHs in the sediment samples was low to moderate-high level, whereas the total health risk through ingestion and dermal contact was considerably high. The PAHs biota sediment accumulation factors data for short-neck clam were obtained in this study, indicating a preferential accumulation of lower molecular weight PAHs. The source apportionment of PAHs in sediment using positive matrix factorization model indicated that the highest contribution to the PAHs was from diesel emissions (30.38%) followed by oil and oil derivate and incomplete coal combustion (23.06%), vehicular emissions (16.43%), wood combustion (15.93%), and natural gas combustion (14.2%). A preliminary evaluation of human health risk using chronic daily intake, hazard index, benzo[a]pyrene-equivalent (BaPeq) concentration, and the incremental lifetime cancer risk indicated that PAHs in short-neck clam would induce potential carcinogenic effects in the consumers.
  10. Zhu F, Storey S, Ashaari MM, Clipson N, Doyle E
    Environ Sci Pollut Res Int, 2017 Feb;24(6):5404-5414.
    PMID: 28025788 DOI: 10.1007/s11356-016-8251-3
    Benzo(a)pyrene degradation was compared in soil that was either composted, incubated at a constant temperature of 22 °C, or incubated under a temperature regime typical of a composting process. After 84 days, significantly more (61%) benzo(a)pyrene was removed from composted soil compared to soils incubated at a constant temperature (29%) or at composting temperatures (46%). Molecular fingerprinting approaches indicated that in composted soils, bacterial community changes were driven by both temperature and organic amendment, while fungal community changes were primarily driven by temperature. Next-generation sequencing data revealed that the bacterial community in composted soil was dominated by Actinobacteria (order Actinomycetales), Firmicutes (class Bacilli), and Proteobacteria (classes Gammaproteobacteria and Alphaproteobacteria), regardless of whether benzo(a)pyrene was present or not. The relative abundance of unclassified Actinomycetales (Actinobacteria) was significantly higher in composted soil when degradation was occurring, indicating a potential role for these organisms in benzo(a)pyrene metabolism. This study provides baseline data for employing straw-based composting strategies for the removal of high molecular weight PAHs from soil and contributes to the knowledge of how microbial communities respond to incubation conditions and pollutant degradation.
  11. Zhang H, Ramamoorthy A, Rengarajan T, Iyappan P, Alahmadi TA, Wainwright M, et al.
    J Biochem Mol Toxicol, 2024 Jan;38(1):e23578.
    PMID: 37927152 DOI: 10.1002/jbt.23578
    Lung cancer is one of the most common cancers in men. Although many diagnostic and treatment regimens have been followed in the treatment for lung cancer, increasing mortality rate due to lung cancer is depressing and hence requires alternative plant based therapeutics with with less side-effects. Myrtenol exhibits anti-inflammatory and antioxidant properties. Hence we intended to study the effect of Myrtenol on B(a)P-induced lung cancer. Our study showed that B(a)P lowered hematological count, decreased phagocyte and avidity indices, nitroblue tetrazolium (NBT) reduction, levels of immunoglubulins, antioxidant levels, whereas Myrtenol treatment restored them back to normal levels. On the other hand, xenobiotic and liver dysfunction marker enzymes and pro-inflammatory cytokines were elevated on B(a)P exposure, which retuned back to normal by Myrtenol. This study thus describes the immunomodulatory and antioxidant effects of Myrtenol on B[a]P-induced immune destruction.
  12. Karami A, Syed MA, Christianus A, Willett KL, Mazzeo JR, Courtenay SC
    J Hazard Mater, 2012 Jul 15;223-224:84-93.
    PMID: 22608400 DOI: 10.1016/j.jhazmat.2012.04.051
    In this study we sought to optimize recovery of fluorescent aromatic compounds (FACs) from the bile of African catfish (Clarias gariepinus) injected with 10mg/kg benzo[a]pyrene (BaP). Fractions of pooled bile were hydrolyzed, combined with ten volumes of methanol, ethanol, acetonitrile, or acetone, centrifuged and supernatants were analyzed by high-performance liquid chromatography with fluorescent detection (HPLC/FL). As well, to test whether FACs were being lost in solids from the centrifugation, pellets were resuspended, hydrolyzed and mixed with six volumes of the organic solvent that produced best FAC recovery from the supernatant, and subjected to HPLC/FL. Highest FAC concentrations were obtained with 2000μl and 1250μl acetone for supernatants and resuspended pellets respectively. FACs concentrations were negatively correlated with biliary protein content but were unaffected by addition of bovine serum albumin (BSA) followed by no incubation indicating that the presence of proteins in the biliary mixture does not simply interfere with detection of FACs. In another experiment, efficiency of acetone addition was compared to two different liquid-liquid extractions (L-LEs). Acetone additions provided significantly higher biliary FACs than the L-LE methods. The new two-stage bile preparation with acetone is an efficient, inexpensive and easily performed method.
  13. Karami A, Keiter S, Hollert H, Courtenay SC
    Environ Sci Pollut Res Int, 2013 Mar;20(3):1586-95.
    PMID: 22752811 DOI: 10.1007/s11356-012-1027-5
    This study represents a first attempt at applying a fuzzy inference system (FIS) and an adaptive neuro-fuzzy inference system (ANFIS) to the field of aquatic biomonitoring for classification of the dosage and time of benzo[a]pyrene (BaP) injection through selected biomarkers in African catfish (Clarias gariepinus). Fish were injected either intramuscularly (i.m.) or intraperitoneally (i.p.) with BaP. Hepatic glutathione S-transferase (GST) activities, relative visceral fat weights (LSI), and four biliary fluorescent aromatic compounds (FACs) concentrations were used as the inputs in the modeling study. Contradictory rules in FIS and ANFIS models appeared after conversion of bioassay results into human language (rule-based system). A "data trimming" approach was proposed to eliminate the conflicts prior to fuzzification. However, the model produced was relevant only to relatively low exposures to BaP, especially through the i.m. route of exposure. Furthermore, sensitivity analysis was unable to raise the classification rate to an acceptable level. In conclusion, FIS and ANFIS models have limited applications in the field of fish biomarker studies.
  14. LIANG SUN TAN, SAW HONG LOH
    MyJurnal
    Polycyclic aromatic hydrocarbons (PAHs) are hazardous and persistent organic pollutants that usually exist at low concentrations in the environment. In this study, dispersive liquid-liquid microextraction (DLLME) technique coupled with high performance liquid chromatography-fluorescence detection (HPLC-FD) was optimized for the analysis of selected PAHs, namely phenanthrene (PHE), fluoranthene (FLA) and benzo[a]pyrene (BaP) in apple juice. Under the optimal extraction conditions (the mixture of 200 µL of acetone and 50 µL of 1-octanol was applied to extract the selected PAHs for 1 min), the DLLME-HPLC-FD showed excellent linearity over the concentration range of 5 to 200 µg/L for both PHE and FLA, and 0.01 to 5 µg/L for BaP with correlation coefficients, r ≥ 0.9956. The method offered ultra-trace detection of selected PAHs in the range of 0.002 to 0.5 µg/L, and negligible matrix effects in determining selected PAHs with relative recovery average within the range of 92.6 to 109.6% in apple juice. The advantages of applying this method for the extraction of PAHs include rapidity, simple operation, as well as small consumption of organic extraction solvent, which is beneficial for routine analysis.
  15. Keshavarzifard, Mehrzad, Mohamad Pauzi Zakaria, Keshavarzifard, Shahin, Sharifi, Reza
    MyJurnal
    This study investigated distributions, composition patterns, sources and potential toxicity of polycyclic aromatic hydrocarbon (PAHs) pollution in surface sediments from the Kim Kim River and Segget River, Peninsular Malaysia. The samples were extracted using Soxhlet extraction, purified using two-step silica gel column chromatography and then analysed by gas chromatography mass spectrometry (GC-MS). The total PAH concentrations ranged from 95.17 to 361.24 ng g-1 dry weight (dw) and 330.09 to 552.76 ng g-1 dw in surface sediments from the Kim Kim and Segget Rivers, respectively. Source type identification using PAH molecular indices and hierarchical cluster analysis (HCA) indicated that PAHs were mostly of pyrogenic origin, while in some stations petrogenic sources had a significant portion. A PAH toxicity assessment using sediment quality guidelines (SQGs), mean effect range-median quotient (M-ERM-Q), benzo[a]pyrene (BaP) equivalent concentration and BaP toxicity equivalent quotient (TEQcarc) indicated low probability of toxicity for both the Kim Kim and Segget Rivers. Moreover, the human health risk assessment applying Cancer Riskingestion and Cancer Riskdermal indicated that probabilistic health risk to humans via ingestion and dermal pathways from sediments of the Kim Kim and Segget Rivers can be categorised as low-to-moderate risk.
  16. Karami A, Christianus A, Ishak Z, Syed MA, Courtenay SC
    Ecotoxicol Environ Saf, 2011 Sep;74(6):1558-66.
    PMID: 21636131 DOI: 10.1016/j.ecoenv.2011.05.012
    This study investigated the dose-dependent and time-course effects of intramuscular (i.m.) and intraperitoneal (i.p.) injection of benzo[a]pyrene (BaP) on the biomarkers EROD activity, GST activity, concentrations of BaP metabolites in bile, and visceral fat deposits (Lipid Somatic Index, LSI) in African catfish (Clarias gariepinus). Intraperitoneal injection resulted in 4.5 times higher accumulation of total selected biliary FACs than i.m. injection. Hepatic GST activities were inhibited by BaP via both injection methods. Dose-response relationships between BaP injection and both biliary FAC concentrations and hepatic GST activities were linear in the i.p. injected group but nonlinear in the i.m. injected fish. Hepatic EROD activity and LSI were not significantly affected by BaP exposure by either injection route. We conclude that i.p. is a more effective route of exposure than i.m. for future ecotoxicological studies of PAH exposure in C. gariepinus.
  17. Abdull Razis AF, Konsue N, Ioannides C
    Asian Pac J Cancer Prev, 2015;16(7):2679-83.
    PMID: 25854346
    BACKGROUND: Phenethyl isothiocyanate (PEITC), the most comprehensively studied aromatic isothiocyanate, has been shown to act as an anti-cancer agent mainly through modulation of biotransformation enzymes responsible for metabolizing carcinogens in the human body. Humans are often exposed to carcinogenic factors, some of which through the diet, such as polycyclic aromatic hydrocarbon benzo[a]pyrene via the consumption of over-cooked meats. Inhibition of the enzymes responsible for the bioactivation of this carcinogen, for example CYP1A1, the major enzyme required for polycyclic aromatic hydrocarbons (PAHs) bioactivation, is recognized as a chemoprevention strategy.

    OBJECTIVE: To evaluate the inhibitory effects of PEITC against benzo[a]pyrene-induced rise in rat liver CYP1A1 mRNA and apoprotein levels.

    MATERIALS AND METHODS: Precision cut rat liver slices were treated with benzo[a]pyrene at 1 and 5 μM in the presence of PEITC (1-25 μM) for 24 hours, followed by determination of CYP1A1 mRNA and apoprotein levels using quantitative polymerase chain reaction and immunoblotting.

    RESULTS: Findings revealed that PEITC inhibited benzo[a]pyrene-induced rise in rat liver CYP1A1 mRNA in a dose-dependent manner as well as the apoprotein levels of CYP1A.

    CONCLUSIONS: It was demonstrated that PEITC can directly inhibit the bioactivation of benzo[a]pyrene, indicating chemopreventive potential.

  18. Latif IK, Karim AJ, Zuki AB, Zamri-Saad M, Niu JP, Noordin MM
    Poult Sci, 2010 Jul;89(7):1379-88.
    PMID: 20548065 DOI: 10.3382/ps.2009-00622
    Aftermath in several air pollution episodes with high concentrations of polycyclic aromatic hydrocarbons did not significantly affect health and performance of broilers despite its renowned sensitivity to polycyclic aromatic hydrocarbons. The aim of the study was to elucidate the previous lack of response in birds exposed to such severe episodes of air pollution. Benzo[a]pyrene (BaP) was used to simulate the influence of air pollution on hematology, selected organ function, and oxidative stress in broilers. One-day-old chicks were assigned to 5 equal groups composed of a control group, tricaprylin group, and 3 groups treated with BaP (at 1.5 microg, 150 microg, or 15 mg/kg of BW). The BaP was intratracheally administered to 1-d-old chicks for 5 consecutive days. The hematology, liver and kidney function, P450 activity, and malondialdehyde level especially in the group receiving 15 mg of BaP/kg of BW demonstrated evidence of hemato- and hepatoxicity via BaP-induced oxidative stress. The deleterious effect of exposure to high concentration of BaP in broiler chickens was probably due to the anatomy of this species and the half-life of BaP. Although the effect of BaP may be transient or irreversible, pathogen challenges faced during the period of suppression may prove fatal.
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