146 



FISHERY BULLETIN OF THE FISH AND WILDLIFE SERVICE 



serum protein content, and elect rophoretic pat- 

 tern wei-e made. Tlie sex and stage of gonad 

 development of each fish were recorded when the 

 blood sample was taken. 



DETERMINATIONS OF BLOOD PROPERTIES 



Hemoglobin Content 



Detennination of the hemoglobin content of in- 

 dividual citrated blood samples was made with the 

 cyanmethemoglobin method, using reagent and 

 standard supplied by Hycel Homione Chemistry 

 Laboratory, Houston, Tex. With a Salili pipette, 

 .02 ml. blood was added to 5 ml. cyanmethemoglo- 

 bin reagent and thoroughly mixed. Contents were 

 transferred after 15 minutes to a cuvette and read 

 colori metrically with a Photovolt Lumetron 

 colorimeter at a wavelength of 530 millimicrons. 

 The colorimeter reading was transferred to a 

 standard hemoglobin curve and hemoglobin con- 

 centration obtained in grams per 100 ml. 



Sedimentation Rate 



The rate of settling of erythrocj^es was de- 

 termined with a standard Westergren blood sedi- 

 mentation apparatus. Individual samples of 

 citrated blood was drawn into Westergren pipettes 

 to the 100-millimeter mark and placed vertically 

 in a Westergren rack. The number of millimeters 

 that the erythrocytes had dropped at the end of 1, 

 2, and 3 houre — the actual sedimentation — was re- 

 corded and multiplied by 2 to make results com- 

 parable with standard tests that use the entire 200- 

 mm. pipette length. Results are expressed in 

 terms of the standard 200-mm. length. 



Erythrocyte Fragility 



Sodium chloride solutions ranging from 0.3 to 

 1.5 percent in 0.1-percent increasing steps were 

 used to test erythrocyte fragility. For each 

 sample, .05 ml. of a 50-percent cell suspension was 

 added to 1 ml. of each saline dilution and readings 

 of "no hemolysis," "partial hemolysis;," or "com- 

 plete hemolysis" were recorded for each tube at 

 the end of 1 hour's incubation at 4° C. 



Total Serum Proteins 



The biuret method of Kingsley (1942) was 

 used to determine total serum proteins. This in- 

 volves precipitation with acetone-alcohol followed 

 by addition of biuret reagent. Readings wei-e 



made with a Photovolt Lumetron colorimeter at a 

 wave length of 530 millimicrons and were plotted 

 against a standard curve prepared with dilutions 

 of clinical chemistry control senmi, supplied by 

 Hyland Laboratories, Los Angeles, Calif., to ob- 

 tain total protein values. 



Serum Chlorides 



Determinations of serum chlorides were made 

 by the standard method outlined in the manual for 

 Photovolt Lumetron colorimeter. This involves 

 treatment of serum with tungstic acid, silver 

 iodate, phosphoric acid, and potassium iodide. 

 Readings were made with the Lumetron colorim- 

 eter at a wave length of 420 millimicrons and 

 plotted against a standard curve prepared from 

 known chloride concentrations to obtain serum 

 chloride values. 



Serum Electrophoresis 



Elect rophoretic examination of serum was made 

 with a Spinco paper electrophoresis system. 

 Samples were run for 6 hours at 15 milliam- 

 peres, with veronal buffer of pH 8.6 and ionic 

 strength of .05. Pooled human serum was used 

 as a standard with each series. Filter-paper strips 

 were dyed with bromphenol blue and analysed 

 with a Spinco Analytrol densitometer. 



COMPARISON OF BLOOD PROPERTIES 



Tests of serum and cellular components of the 

 blood of prespawning and postspawning alewives 

 pro\aded the data presented in table 1. Results 

 from the two Maine spawning nms haA'e been com- 

 bined, since no consistent differences between them 

 were noted. 



HEMOGLOBIN 



Variations in hemoglobin content of fish blood 

 have been examined in a number of marine and 

 fresh-water species by several investigators. 

 Black (1955) found that the hemoglobin level of 

 largemouth bass increased after forced exercise, 

 but that of five other fish species did not. Pavlov 

 and Krolik (1936) found that hemoglobin in- 

 creased with the ripening of the sex products, 

 while Naumov (1956) noted that it increased to 

 the time of spawning and then dropped to a very 

 low level. Gelineo (1957) found that hemoglobin 

 values for several species of marine fish were some- 



