FISHERY BULLETIN: VOL. 73, NO. 2 



A series of measurements of 100 spines was 

 made with an ocular micrometer at a magnifica- 

 tion of 30 X. At tiiis magnification, one ocular 

 micrometer unit equals 0.033 mm. The radius was 

 measured at midspine on a line perpendicular to 

 and beginning at the indentation axis. Distances 

 from the center of the spine to each annulus were 

 recorded for back calculation of length, along the 

 radius line. Finally, the distance from the spine 

 margin to the outermost annulus was measured 

 for determination of the time of annulus forma- 

 tion. 



Methods of Reproductive Biology 



In the laboratory, each gonad was blotted dry, 

 weighed, and a segment of one lobe was 

 dehydrated in alcohol and embedded in paraffin. 

 Slides were prepared of cross-sections of the lobe, 

 cut at thicknesses of 5, 10, and 25 ^m; thicker sec- 

 tions have less tendency to collapse and were made 

 to ensure that the overall configuration of the 

 cross-section could be observed. Sections were 

 stained with ehrlich's hematoxylin and eosin. 



Each gonad was classified according to sex and 

 state of development. Assignment of a develop- 

 mental class depended on the predominate stage 

 of gametogenesis seen in the gonad. The division 

 of gametogenic stages is as follows: 



Oogenesis was divided into five stages, follow- 

 ing criteria detailed for a variety of species by 

 Kraft and Peters (1963). Smith (1965), and Moe 

 (1969). 



Stage 1. Very small (15-30 /xm in diameter) 

 oocytes with a large nucleus, single nucleolus, and 

 a relatively small amount of basophilic cytoplasm. 



Stage 2. (30-50 ju,m) Previtellogenic oocytes with 

 a strongly basophilic cytoplasm and multiple 

 nucleoli around the nucleus margin. 



Stage 3. (150-300 [xm) Vitellogenesis begins 

 with the deposition of yolk vesicles in the less 

 darkly staining cytoplasm. A thin zona radiata can 

 be seen in late stage 3. 



Stage 4. (280-450 /^m) Cytoplasm filled with yolk 

 vesicles and globules; the zona radiata well 

 developed and strongly acidophilic. 



Stage 5. (450-1,050 ixm) Mature or nearly ma- 

 ture oocytes, uniform in appearance due to the 

 coalescence of the yolk globules. The nucleus is 

 eccentric and the zona radiata is thin and non- 

 striated. These oocytes are often extremely 

 irregular in outline and Roede (1972), who noted 



the same irregularity in mature eggs of other 

 labrids, probably correctly attributed this to dis- 

 tortion during fixation and staining. This stage 

 was seldom seen, but several specimens were seen 

 with eggs in the ovarian lumen and stage 5 oocytes 

 still within the follicle. 



Spermatogenesis occurs in small crypts, in 

 which all the cells are at the same stage. The 

 development and appearance of the sperma- 

 togonia, primary spermatocytes, secondary sper- 

 matocytes, spermatids, and mature sperm follows 

 very closely the descriptions given by Hyder (1969) 

 for Tilapia and by Moe (1969) for Epinephelus 

 morio, and will not be repeated here. 



The gonadal development classes, intended to 

 parallel those of Moe (1969) and Smith (1965), were 

 designated as follows: 



Class 1. Immature female. Stages 1 and 2 

 oocytes present, atretic or brown bodies (Chan et 

 al. 1967) absent. The ovarian lamellae are pressed 

 closely together and the lumen is small. 



Class 2. Resting mature female. Oocyte stages 

 1, 2, and 3 present, with stage 2 predominating. 

 Atretic bodies are usually present. 



Class 3. Active mature female. Oocyte stages 3 

 and 4 predominate in the lamellae. In late class 3, 

 stage 5 oocytes are also present. 



Class 4. Postspawning female. Ovary is 

 disrupted, with many empty follicles in the 

 lamellae. Some degenerating stages 4 and 5 

 oocytes are usually found in the lamellae and lumen, 

 respectively. 



Class 5. Transitional. Seminiferous crypts 

 begin to proliferate in the lamellae, but some stage 

 2 oocytes can be seen. These oocytes degenerate 

 and decrease in number as spermatogenic activity 

 begins to dominate the gonad. 



Class 6. Inactive male. Crypts containing 

 primary and secondary spermatocytes pre- 

 dominate; few spermatids and mature sperm are 

 seen. 



Class 7. Active male. Spermatids and tailed 

 sperm increase in abundance until, in the ripe 

 phase, sperm are densely packed in the collecting 

 ducts and many crypts have coalesced. 



Class 8. Postspawning male. Ducts are still ex- 

 panded, but few sperm can be seen in them. Many 

 new crypts containing spermatogonia are present. 

 This apparently is a short-lived stage that rapidly 

 gives way to the resting (class 6) testis. 



Fecundity determinations were made by count- 



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