EFFECTS OF BENZO(A)PYRENE ON THE EARLY DEVELOPMENT 

 OF CALIFORNIA GRUNION, LEURESTHES TENUIS 

 (PISCES, ATHERINIDAE) 



Delaine L. Winkler, 1 Keith L. Duncan, 2 Jo Ellen Hose,' and Harold W. Puffer 4 



ABSTRACT 



Benzo(a)pyrene (BaP), which is carcinogenic and mutagenic in mammals, exists worldwide in the marine 

 environment. Sources of this polycyclic aromatic hydrocarbon include oil spills, industrial effluents, and 

 atmospheric fallout. This study is the first to examine the effects of BaP on the embryonic development of a 

 teleost, the California grunion. Gametes were stripped from spawning adults, and eggs were artificially fer- 

 tilized. The fertilized eggs were then incubated for up to 14 days with initial concentrations of BaP ranging 

 from to 869 ppb. Steady-state tissue levels of BaP ranged from 0.46 to 19.92 ppm, which represented 

 bioaccumulation factors of 146-437 times the steady-state BaP concentrations in seawater. When compared 

 with controls, embryos exposed to initial BaP levels of 24 ppb or greater showed decreased hatchings, 

 reduced notochord lengths, and increased morphological abnormalities. These results suggest that exposure 

 of grunion embryos to BaP in contaminated areas may lead to their decreased survival. 



Benzo(a)pyrene (BaP) is one oil constituent com- 

 monly found in marine sediments and organisms 

 (ZoBell 1971; Neff 1979). This polycyclic aromatic 

 hydrocarbon is introduced into the ocean via oil 

 spillage, offshore drilling leaks, industrial effluents, 

 runoff of asphalt roads, creosoted pilings, and at- 

 mospheric fallout (Andelman and Suess 1970; Dunn 

 1976; Puffer et al. 1979). Because BaP exhibits toxic, 

 mutagenic, and carcinogenic properties in mammals, 

 one might infer that it could also exert detrimental ef- 

 fects on fish populations (Heidelberger 1975; Miller 

 1978). Such effects could lead directly to a decrease 

 in a valuable food source and pose a public health 

 problem in the consumption of contaminated 

 seafoods (Dunn and Fee 1979). 



Awareness of significant BaP contamination in the 

 marine biota has led to research on adult stages (Lee 

 etal. 1972; Puffer etal. 1979) and, more recently, the 

 sensitive embryonic-larval stages of fish (Hose et al. 

 1981, 1982). Grunion are particularly suitable for 

 such a study because their embryonic development is 

 well documented and they are easily reared in cap- 



'Department of Biological Sciences, University of Southern 

 California, University Park, Los Angeles. Calif.; present address: 

 Environmental Management Division, Port of Los Angeles, 425 S. 

 Palos Verdes Street. San Pedro, CA 90733. 



2 Department of Pathology, School of Medicine, University of 

 Southern California, Los Angeles, Calif; present address: Depart- 

 ment of Pathology, School of Medicine, University of California at 

 San Francisco, San Francisco, CA 94143. 



'Department of Pathology, School of Medicine, University of 

 Southern California, Los Angeles, CA 90033, and VANTUNA 

 Research Group, Occidental College, Los Angeles, CA 90041. 



■"Department of Pathology, School of Medicine, University of 

 Southern California, Los Angeles, CA 90033. 



Manuscript accepted November 1982. 

 FISHERY BULLETIN: VOL. 81, NO. 3, 1983. 



tivity (David 1939; Ehrlich and Farris 1971). 

 Furthermore, grunion spawn on sandy beaches 

 where developing eggs remain in the sand until the 

 tide uncovers, agitates, and stimulates the eggs to 

 hatch (Walker 1952). During this time, the develop- 

 ing eggs may be exposed to BaP. Therefore, we have 

 undertaken this study to examine the effects of BaP 

 on the early life history of California grunion, 

 Leuresthes tenuis. 



MATERIALS AND METHODS 



Decontaminated seawater (sterilized, free of detect- 

 able BaP and particulate matter) was obtained by ex- 

 posure of Los Angeles Harbor water to direct 

 sunlight for 1 wk. Photooxidation by sunlight 

 resulted in the breakdown of contaminating BaP to 

 noncarcinogenic byproducts such as phenols and 

 quinones (National Academy of Sciences 1972). 

 Seawater exposed to sunlight was filtered through 

 Whatman No. 5 filters to remove large, particulate 

 matter and then sterilized by ultraviolet light. Water 

 was maintained at a salinity of 31-32 %o, a pH of 7.7- 

 7.9, and a temperature of 20.0°-21.5°C. 



BaP was dissolved in acetone, mixed with decon- 

 taminated seawater, and stirred for 24 h. The added 

 concentrations of BaP in seawater were 5, 10, 100, 

 500, 1,000, and 5,000 ppb. The final concentration of 

 acetone in the control and BaP-treated groups did 

 not exceed 0.04 9< . Spawning California grunion were 

 collected at Redondo Beach, Calif. Gametes were 

 stripped and artificially fertilized in BaP-free, decon- 



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