METHODS 



Eggs from ripe fish collected in Bellingham Bay, 

 Wash., were artificially fertilized on 12 March 

 1974 and were reared at the Mukilteo field station 

 of the Northwest and Alaska Fisheries Center 

 (NWAFC), National Marine Fisheries Service 

 (NMFS), NOAA, Seattle, Wash. The eggs were 

 incubated in 0.6 or 1.0 1 glass beakers containing 

 local Puget Sound seawater maintained at 9°-10° 

 C. Subsamples of eggs were preserved in Forma- 

 lin^ in eight time periods, 4, 9, 20, 24, 48, 72, 96, 

 and 120 h, after fertilization. Subsamples of larvae 

 were preserved for each of 13 time periods: at 

 hatching (144 h or 6 d after fertilization) and at 

 2, 3, 4, 7, 9, 11, 14, 16, 18, 21, 23, and 25 d 

 after hatching. Although rotifers [Brachionus 

 plicatilus) were added to the containers at various 

 levels (4, 8, and 16 rotifers/ml seawater), the lar- 

 vae did not feed actively and all were dead by 22 

 April 1974. 



Approximately 300 larvae were obtained from 

 NWAFC collections made off coastal Washington 

 in 1972. An additional 107 larvae were obtained 

 from Oregon State University (OSU) ichthyo- 

 plankton collections taken off the Oregon coast 

 from 1971 to 1972 (Richardson and Pearcy 1977; 

 Richardson see footnote 4). Benthic juveniles were 

 collected in beam trawls off the mouth of the 

 Columbia River in June and September 1975 

 (Richardson et al.^). 



Counts of meristic structures were made on 209 

 larvae (3.2-23.6 mm) and 7 juveniles (44-160 mm) 

 taken from the OSU and NWAFC collections. 

 Larvae were stained with Alizarin Red S using 

 Taylor's (1967) enzyme method to determine 

 sequence of ossification. Some (45) were sub- 

 sequently restained with Alcian Blue and Alizarin 

 Red using techniques described by Dingerkus and 

 Uhler ( 1977). Counts were made of dorsal fin rays, 

 anal fin rays, caudal fin rays, left and right pec- 

 toral fin rays, branchiostegal rays, gill rakers, ver- 

 tebral centra, neural spines, and haemal spines. 

 Fin rays and vertebrae were counted even if they 

 were tinted only slightly with alizarin stain. Up- 

 take and retention of alizarin in ossified structures 

 may vary depending upon the length of time the 



''lieference to trade names does not imply endorsement by the 

 National Marine Fisheries Service, NOAA. 



sRichardson, M. D., A. G. Carey, Jr., W. A. Col- 

 gate. 1977. The effects of dredged material disposal on 

 benthic assemblages off the mouth of the Columbia River. Fi- 

 nal Rep., Dep. Army Corps Eng. Contracts DACW 57-75-C-0137 

 and DACW 57-56-C-0092, Vicksburg, Miss., 411 p. 



FISHERY BULLETIN: VOL. 78, NO. 2 



specimens have been in preservative. Differential 

 loss of stain may account for some of the variation 

 observed in the onset of ossification of certain 

 structures such as teeth. 



Measurements were made using an ocular mi- 

 crometer in a stereomicroscope. The greatest out- 

 side diameter and greatest yolk diameter were 

 recorded for 80 eggs from the reared series, 10 for 

 each of the eight time periods that eggs were pre- 

 served from 4 to 120 h after fertilization. Mea- 

 surements were made on 63 reared larvae, 5 

 specimens (when available) from each of the 13 

 time periods after hatching (6 d after fertilization) 

 that larvae were preserved to 25 d after hatching. 

 Size range of reared specimens was 2.7-5.3 mm SL 

 (standard length). Measurements also were made 

 on 107 larvae from plankton collections including 

 5 specimens (when available) for each 1.0 mm size 

 class interval from 2 to 23 mm SL (range 2.9-23.6 

 mm). Body measurements were made on larvae as 

 follows: 



Standard length - snout tip to notochord tip 

 until notochord is fully flexed and the poste- 

 rior margin of the forming hypural elements 

 is vertical, then to posterior margin of hypur- 

 als. 



Head length - snout tip to cleithrum. 



Body depth at pectoral fin base - vertical dis- 

 tance from dorsal body margin to ventral body 

 margin, excluding finfold or fins, at the pec- 

 toral fin base. 



Body depth at anus - vertical distance from 

 dorsal body margin, excluding finfold or fin, to 

 anus. 



Body depth behind anus - vertical distance from 

 dorsal body margin to ventral body margin, 

 excluding finfolds or fins, at point im- 

 mediately behind anus where body depth de- 

 creases greatly compared to depth at anus. 



Body depth at caudal peduncle - before forma- 

 tion of caudal fin, vertical distance from dor- 

 sal body margin to ventral body margin, 

 excluding finfolds or fins, at the posteriormost 

 myomere; after caudal fin formation, least 

 depth of caudal peduncle. 



Snout length - snout tip to anterior margin of 

 right eye. 



Eye diameter - horizontal distance across right 

 eyeball. 



Snout to anus length - distance along body mid- 

 line from snout tip to vertical through posteri- 

 or margin of anus. 



402 



