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Preliminary study on adverse effects of phenanthrene and its methyl and phenyl derivatives in larval zebrafish, Danio rerio

Wolińska L., Brzuzan P., Woźny M., Góra M., Łuczyński M. K., Podlasz P., Kolwicz S., Piasecka A.

Environmental Biotechnology

2011

Vol. 7, No. 1

26-33

Toxic effects of polycyclic aromatic hydrocarbons (PAHs) have been extensively studied in fish, although knowledge concerning biological activities of phenanthrene and its derivatives remains still incomplete. The aim of this work was to evaluate lethal and sublethal effects of benzo(a)pyrene, phenanthrene and phenanthrene derivatives (1-methylphenanthrene, 4-methylphenanthrene, 1-phenylphenanthrene and 4-phenylphenanthrene) on zebrafish (Danio rerio) larvae. We conducted acute toxicity test, using 96h static renewal exposure to a series of the PAH concentrations (0.05, 0.50, 5.00, 50.00µmol*l-1), to determine the No Effect Concentration (NEC) value for each compound examined. The mean NEC estimates obtained in the study were 5.16۪.45µmol*l-1 (B[a]P), 4.88۪.13µmol*l-1 (Ph), 40.24䔰.93µmol*l-1 (1P-Ph), 47.92ۭ.61µmol*l-1 (1M-Ph), 24.31۱.33µmol*l-1 (4P-Ph) and 3.11۫.01µmol*l-1 (4M-Ph) and suggested the following order of PAH toxicities on Danio rerio larvae: 4M-Ph>Ph˜B[a]P>4PPhᲙP-Ph>1M-Ph. To gain insight into possible molecular mechanisms of apparent toxicity of phenanthrene derivatives on zebrafish larvae, we examined mRNA expression of cyp1a, cyp1b1, and vtg genes in the larvae exposed for 48h to a PAH concentration of 0.50µmol*l-1. Whereas the larvae exposed to each tested PAH displayed many developmental abnormalities (i.e. pericardial and yolk sac edema, dorsal curvature, or tail malformations), no significant upregulation of cyp1a and cyp1b1 mRNA was observed, except for zebrafish exposed to B[a]P. However, significant reduction of vtg mRNA was observed in the larvae exposed to phenanthrene and its 4P- derivative. The results may contribute to the development of a new knowledge about effects of structurally diverse phenanthrene derivatives on vertebrate organisms.

Cyclopenta[c]phenanthrene induction of CYP1A in brain of rainbow trout (Oncorhynchus mykiss)

Brzuzan P., Woźny M., Łuczyński M. K., Góra M., Ciesielski S., Kuźmiński H.

Environmental Biotechnology

2007

Vol. 3, No. 1

10-14

We assessed the effects of cyclopenta[c]phenanthrene (CP[c]Ph) and benzo[a]pyrene (B[a]P; positive control) on CYP1A gene expression in brain of juvenile rainbow trout (Oncorhynchus mykiss) using the quantitative reverse transcription polymerase chain reaction (Q-RT-PCR). A group of hatchery raised rainbow trout, with an average body mass of 49.4 g and total length of 15.5 cm were given an intraperitoneal injection (10 mg*kg-1) of either CP[c]Ph or B[a]P in corn oil (2 mg*mi-1 corn oil) or corn oil alone (control). After 24 and 48 h, trout brains were collected for mRNA isolation and analysis. After 24 hours of the exposure, only B[a]P-treated rainbow trout had 10-fold higher number of CYP1A transcripts (mean = 3.63*106 transcripts*µg-1 total RNA) than control fish (3.24*105 transcripts*µg-1 total RNA; Tukey test, P<0.05). After 48 hrs, significantly higher levels of CYP1A expression (Tukey test, P<0.001) were found in either CP[c]Ph- or B[a]P- induced group (1.45*106 and 6.92*106 transcriptsźµg-1 total RNA, respectively) over a control group (mean=1.41*105 transcripts*µg-1 total RNA). The finding that CYP1A in brain tissue was inducible by CP[c]Ph, a polycyclic aromatic hydrocarbon (PAH) of different than B[a]P planar characteristics, may further validate the use of rainbow trout brain CYP1A mRNA levels as a biomarker of PAH exposure.