Birds are potentially vulnerable to the toxicity of certain environmental pollutants due to limited detoxification capabilities of their liver microsomal cytochrome P450 (CYP) enzymes. In wild birds, ethoxyresorufin O-deethylation (EROD) activity, a marker of CYP1A activity in mammals and domestic chickens, has been used as a biomarker of exposure to polychlorinated biphenyls (PCBs), dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs). The aim of the present study was to investigate hepatic CYP activity as an indication of detoxification capacity in New Zealand birds. In addition to the use of conventional in vitro CYP activity assays, the applicability of a noninvasive CYP activity assay was tested using caffeine as the in vivo substrate.
The ontogeny of liver microsomal 3-hydroxylation of quinine, a marker of human CYP3A activity, was investigated in Adelie penguins (Pygoscelis adeliae) from Ross Island, Antarctica. The results indicate that chicks (2-4 weeks old) possess a CYP3A-like isoform(s) as active as but not identical to the CYP3A-like isoform(s) in adults. Total CYP content was low at 2 weeks of age and increased rapidly and linearly approaching adult levels by 4 weeks of age implying a rapid development of CYPs other than the CYP3A-like isoform(s).
The main study was conducted on adult (and some post-fledging immature) birds of two native species, the herbivorous paradise shelduck (Tadorna variegata) and the omnivorous southern black-backed gull (Larus dominicanus). Birds were shot for liver collection at three sites in the South Island of New Zealand; West Coast, Lake Waipori and Dunedin landfill, in 2001-2002. The results indicate that shelducks posssess multiple CYP isoforms that independently catalyse EROD, p-nitrophenol hydroxylation (p-NP) and erythromycin demethylation (EMD), markers of mammalian CYP1A, CYP2E and CYP3A activity, respectively. In contrast, gulls appear to possess a single isoform catalysing both EROD and p-NP but possess no isoform capable of catalysing EMD.
EROD activity was high in shelducks and gulls from the landfill site, although it was not significantly associated with liver concentrations of PCBs (0.079-6.2 and 8.2-310 ng/g in shelducks and gulls, respectively), PCDD/PCDFs, toxic equivalents (TEQs) and dichlorodiphenyldichloroethylene (DDE) (0.85-317 and 44-4800 ng/g in shelducks and gulls, respectively) in either species. In shelduck livers from the landfill site, EROD was positively associated with Pb concentration but negatively associated with Hg concentration. Assessment of PCB congener patterns based on concentration ratios of individual congeners to the reference congener, 2,2�,4,4�,5,5�-hexachlorobiphenyl (IUPAC #153), indicate that the metabolism of 2,4,4�-trichlorobiphenyl (PCB#28) and 2,4,4�,5-tetrachlorobiphenyl (PCB#74) is inducible in shelducks but not in gulls. Hepatic reduced glutathione (GSH) content was higher in gulls than in shelducks suggesting greater resistance to oxidative stress in gulls.
The in vivo caffeine metabolism test as a noninvasive method to determine CYP1A activity in shelducks and gulls gave a positive outcome. The test was performed by administration of a single intraperitoneal dose of caffeine (1 mg/kg body weight) followed by blood collection at 2 and 4 h after caffeine administration for determination of the serum concentration ratio of the metabolite, paraxanthine, to caffeine (PX/CA) by HPLC. In both species, the PX/CA ratio was markedly increased by pretreatment with the model CYP1A inducer, β-naphthoflavone (BNF). BNF treatment also increased EROD activity determined after death (80-fold and 20-fold compared to controls in shelducks and gulls, respectively). However, sensitivity of the PX/CA ratio approach was lower in gulls than in shelducks due presumably to the formation of unidentified caffeine metabolites in gulls. Immunoblot analysis failed to reveal increased CYP protein levels caused by BNF treatment in shelducks and gulls due to poor cross-reactivity of avian proteins with polyclonal antibodies raised against mammalian CYPs.
EROD activity was also determined in livers of the piscivorous yellow-eyed penguin (Megadyptes antipodes) (1 chick, 3 post-fledging immature, 1 adult) from Otago, South Island of New Zealand, and found to be below the limit of quantitation. The adult liver contained 18.5 ng/g of total PCBs suggesting that EROD in this species is insensitive to induction. Comparison of the PCB congener pattern based on [PCBx]/[PCB#153] between the penguin and its putative source of PCB exposure, New Zealand marine fish, indicates that CYPs in yellow-eyed penguins metabolise 2,2�,5,5�-tetrachlorobiphenyl (PCB#52) and 2,2�,4,5,5�-pentachlorobiphenyl (PCB#101) as in many other avian species.
The findings of this study highlight substantial species differences in CYP activity in wild birds. Whether CYP expression in New Zealand birds is genetically distinct from birds in other parts of the world may warrant further investigation.
| Identifer | oai:union.ndltd.org:ADTP/217583 |
| Date | January 2006 |
| Creators | Numata, Mihoko, n/a |
| Publisher | University of Otago. School of Pharmacy |
| Source Sets | Australiasian Digital Theses Program |
| Language | English |
| Detected Language | English |
| Rights | http://policy01.otago.ac.nz/policies/FMPro?-db=policies.fm&-format=viewpolicy.html&-lay=viewpolicy&-sortfield=Title&Type=Academic&-recid=33025&-find), Copyright Mihoko Numata |
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