Table 1. — Aniisera iiscrl in this stiidij 



parts 1 percent saline in a Waring Blender. - 

 After centrifugation, the addition of 11 parts 

 of distilled water precipitated the vitellins 

 from the supernatant fluid. The precipitate 

 was dissolved again in saline, reprecipitated 

 and redissolved, and used for injections. Whole 

 serum from a mature female Pacific cod was 

 u.sed to produce the anticod vitellin reagent. 

 The resulting antiserum was absorbed at a 1 :1 

 ratio with male cod serum before testing. 

 Usually the vitellin-bearing materials were 

 suspended in a bayol-arlacel mixture and in- 

 jected into the rabbits intraperitioneally. Con- 

 sistently uniform results were obtained when 

 other injection procedures were used, but a 

 greater number of injections was usually re- 

 quired. Single bleedings were used for testing 

 with the exception of the reagent prepared 

 against starry flounder vitellin which was a 

 pool of numerous bleedings from five rabbits. 

 The antisera produced in different rabbits in- 

 jected with the same vitellin material were 

 qualitatively very similar. This uniformity of 

 reagent indicates that the differences reported 

 later are not due to variations in the immune 

 response of individual rabbits. 



COLLECTION OF SERUM SAMPLES 



Samples of fish serum were taken from 

 whole blood that had been processed within 48 

 hours after collection ; the samples were then 

 stored at -35° C, a temperature at which the 

 vitellin fraction appeared to be stable. Some 

 sera had been stored as long as 8 years when 

 tested. 



REACTIVE PROPERTIES OF THE 

 ANTISERA AND VITELLINS TESTED 

 Table 2 summarizes the data obtained 



l: Trade names referred lo in lliis piibli<-atioii do not imply 

 en«Iorsement »»f coMilriercia! iinidui-ts. 



through testing of sera from mature females of 

 various fish species. All antisera were tested 

 with the same fish sera ; this testing included 

 males as well as females from most species. 

 The only reaction with male serum occurred 

 between the antirockfish reagent and male rock- 

 fish serum. This reaction was very weak and 

 was most likely the result of nonvitellin anti- 

 gens present in the injected material. The re- 

 action with male rockfish sera could not be 

 confused with the reaction with female rockfish 

 sera. 



Table 2. — Cross-reacliriti/ of rabbit antilelcnal vitellin sera tcilh 

 vitellins of fish representing various laxonomic groups ' 



(1) P.'jcific lamprey. Lamprira iTidnlnln: (2) spiny dogfish, Squalus acanthias: 

 (31 White sturgeon. Aripni:^fr trnit^innntmius: (4) sh;id. Alona .yapidissima: 

 Pacific herring, Cliipca hareiigiis pallasi: (.l) sockeye salmon, Oncorhynchus 

 nerkii: (6) carp. Cyprinus carpin: northern sqiiawfish I'lychochrilus oregon- 

 etisis: largescalc sucker, Catostomns macrochfitus; (7) Pacific hake, \ferlucciux 

 prnduclux: Pacific cod. Gadus macrocephaliis: (8) bigeye t(nia, Thuniius 

 oUfnuH; Pacific mackerel. Scomber jopajticux; (9) copper rockfish, Sehaslodes 

 canrinus: (101 cabczon, !icorparniclilhy!i marmnrnlnx: staghorn sculpln, 

 Lcplocotlus armalus; (11) sand sole, I'srilichthys mdnnoxlirliix: English sole, 

 Parophrya veluhis: starry flounder, Plnlkhlhys xirllninx: (12) northern mid- 

 sliipman, Porkhibys notatus. 



All antisera reacted strongly with sera from 

 mature females within the families that pro- 

 vided the vitellin for antibody stimulation. 

 Because of this high degree of cros.s-reactivity 

 within families, the reactions of the antisera 

 may be considered mainly with regard to the 

 family rather than the species from which the 

 vitellin used for antibody stimulation origin- 

 ated. The arbitrary designations given in 

 tables 1 and 2 refer to vitellin of any species of 

 that family. 



Four of the reagents also reacted strongly 

 beyond the immediate family group; two of 



204 



U.S. FISH AND WILDLIFE SERVICE 



