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Craig A. Harms

Publications and source records attributed to Craig A. Harms.

3 recordsLinked to original sources

Anesthesia and liver biopsy techniques for pigeon guillemots ( Cepphus columba ) suspected of exposure to crude oil in marine environments

This paper reports on the anesthesia and liver biopsy techniques used in adult and nestling pigeon guillemots ( Cepphus columba ) to test for continued exposure to residual crude oil in the marine environment. Populations of pigeon guillemots have declined significantly in Prince William Sound, Alaska, USA, possibly because of residual effects of crude oil in the environment after the Exxon Valdez oil spill in March 1989. Measurement of hepatic cytochrome P450 1A (CYP1A) is currently the best way to assess crude oil exposure from food sources; however, lethal sampling to obtain adequate liver tissue was not desirable in this declining population of birds. As part of a larger study to identify factors limiting the recovery of pigeon guillemots and other seabird populations, we surgically collected liver samples from adult and nestling guillemots to provide samples for measurement of hepatic CYP1A concentrations. Results from the larger study were reported elsewhere. Liver samples were taken from 26 nestling (1998) and 24 adult (1999) guillemots from a previously oiled site (Naked Island; 12 chicks, 13 adults) and from a nonoiled site (Jackpot Island/Icy Bay; 14 chicks, 11 adults). The birds were anesthetized with isoflurane. No surgical complications occurred with any of the birds and all adult and nestling birds survived after surgery to the point of release or return to the nest. Thirteen out of 14 chicks from the Jackpot Island/Icy Bay and 8 out of 12 chicks from Naked Island fledged. Four chicks at Naked Island were depredated before fledging. All adults abandoned their nests after surgery, so the study sites were revisited the following summer (2000) in an attempt to assess overwinter survival of the adults. All but 1 adult biopsied bird at the nonoiled site (Icy Bay) was found renesting, whereas only 2 birds at the previously oiled site (Naked Island) were similarly observed. The percent of 1999 breeders at Naked Island that returned to their nest sites to breed again in 2000 was low at nests of biopsied birds and nonbiopsied birds alike, suggesting that factors other than the surgical procedure were responsible for the low return rate among this group. These survival results provide strong support for using experienced veterinarians for nonlethal invasive sample collection from birds to document exposure to crude oil in the marine environment.

Alaska

Quantitative polymerase chain reaction for transforming growth factor-β applied to a field study of fish health in Chesapeake Bay tributaries

Fish morbidity and mortality events in Chesapeake Bay tributaries have aroused concern over the health of this important aquatic ecosystem. We applied a recently described method for quantifying mRNA of an immunosuppressive cytokine, transforming growth factor-β (TGF-β), by reverse transcription quantitative-competitive polymerase chain reaction to a field study of fish health in the Chesapeake Basin, and compared the results to those of a traditional cellular immunoassay macrophage bactericidal activity. We selected the white perch ( Morone americana ) as the sentinel fish species because of its abundance at all of the collection sites. White perch were sampled from Chesapeake Bay tributaries in June, August, and October 1998. Splenic mononuclear cell TGF-β mRNA levels increased and anterior kidney macrophage bactericidal activity decreased, particularly in eastern shore tributaries, from June to August and October. The results of the two assays correlated inversely (Kendall's τ b = -0.600; p = 0.0102). The results indicated both temporal and spatial modulation of white perch immune systems in the Chesapeake Basin, and demonstrated the utility of quantitative PCR for TGF-β as a molecular biomarker for field assessment of teleost fish immune status.

Delaware, Maryland, Pennsylvania, Virginia

Correlation of transforming growth factor-β messenger RNA (TGF-β mRNA) expression with cellular immunoassays in Triamcinolone-treated captive hybrid striped bass

Assessing fish immune status with molecular markers has been hampered by a lack of specific reagents. A quantitative polymerase chain reaction (PCR) method (reverse transcription quantitative–competitive PCR, RT-qcPCR) for measuring transforming growth factor-β (TGF-β) transcription from a broad range of teleost fish has recently been developed. The quantitative PCR now permits monitoring production of this important immunosuppressive cytokine in response to immunomodulating agents and conditions. We examined anterior kidney and spleen mononuclear cells from hybrid striped bass (female striped bass Morone saxatilis × male white bass M. chrysops ) for production of TGF-β messenger RNA (mRNA) in response to administration of the synthetic glucocorticoid triamcinolone. We also compared TGF-β transcription with anterior kidney macrophage bactericidal activity and splenic lymphocyte blastogenesis. Anterior kidney mononuclear cell TGF-β mRNA levels decreased, whereas bactericidal activity increased. Spleen TGF-β mRNA levels did not change significantly, and splenic lymphocyte pokeweed mitogen stimulation index increased in triamcinolone-treated fish. Since triamcinolone is used therapeutically as a suppressive immunomodulator, the enhanced immune functions indicated by the cellular immunoassays were unexpected; however, the inverse response of TGF-β production and macrophage bactericidal activity was consistent with the known relationship between TGF-β and macrophage activation in mammals. Induced immunomodulation in hybrid striped bass was detectable by both traditional cellular immunoassays and the new RT-qcPCR for TGF-β.

Journal of Aquatic Animal Health