A PRACTICAL APPROACH TO THE DESCRIPTION AND IDENTIFICATION

OF SEBASTES LARVAE

A Thesis Presented to the Graduate Faculty of California State University, Hayward

In Partial Fulfillment of the Requirements for the Degree Master of Science in Marine Science

By

Lucy Wold October 1991 ABSTRACT

The eastern North Pacific rockfishes (Sebastes, subfamily Sebastinae, family Scorpaenidae) are a speciose group of economically, recreationally, and ecologically important fishes. Approximately 100 species of rockfishes occur worldwide, with the majority of species concentrating in temperate waters between 34° and 38°N latitude (Barsukov, 1982; Seeb, 1986) . Rockfish larvae <7 mm standard length numerically dominate the ichthyoplankton collected within the California Cooperative Oceanic Fisheries Investigations california Current sampling area (Ahlstrom et al., 1978; Kendall, 1991).

However, recent evolutionary divergence of the genus, large numbers of sister species (Barsukov, 1982; Seeb, 1986; Seeb and Kendall, 1991) and similarities in morphology and pigment patterns have made larval species identifications problematic. Prior to this study, there were no published descriptions of the larval stages of the black and yellow rockfish, S. chrysomelas, and only intraovarian descriptions of blue rockfish, S. mystinus larvae (Wales, 1952; Efremenko and Lisovenko, 1970). During the 1987-88 reproductive season, full-term larvae from field-collected females of these common, nearshore species were cultured at the Monterey

11 Ill

Bay Aquarium in Monterey, California, as source specimens for descriptive analyses. Subsamples of the cultured larvae were used to develop criteria for S. mystinus and S. ch~somelas larvae based on pigmentation patterns. The pigment patterns of cultured larvae were compared to those of Monterey Bay Sebastes larvae from ichthyoplankton collections (Wallace, 1988) and to previously described larvae of species that occur in the Monterey Bay area (Moser et al., 1977; Stahl-Johnson, 1985; Kendall and Lenarz, 1986; Moser and Butler, 1987; Kendall, 1989; Matarese et al., 1989; Moreno, 1990) . Cluster analysis dendograms were generated from the comparisons and are discussed in terms of allelic relationships shown in adult, juvenile and larval Sebastes

(Seeb, 1986; Seeb and Kendall, 1991). ACKNOWLEDGEMENTS

Special thanks to the Monterey Bay Aquarium for providing culture space, husbandry assistance, gravid fishes and water chemistry analyses; Allen Grover of the California Department of Fish and Game for his original computer programs; Dr. Robert Lea and David Ventresca of the California Department of Fish and Game for unpublished data on Sebastes reproductive timing in Monterey Bay; Dr. Ron Hedriclc of the University of California Davis for histology sections and post-mortem analyses; Dr. Mickey Eldridge of the National Marine Fisheries Service Bodega Bay and Diana Watters of California Department of Fish and Game for a memorable trip to the Cordell Banks; Refik Ohrun of Hubbs Sea World for technical advice on rotifer culture; Lynn McMasters of Moss Landing Marine Laboratories for larval illustrations and graphics assistance; my thesis committee and the students, staff and faculty of Moss Landing Marine Laboratories. As I tried to put together a list of personal acknowledgements, I realized how fortunate I was to have the technical support and friendship of such a talented and giving group of people during this project. Thank you all, especially Allie B. and Little Ray.

v VI

This work is a result of research sponsored in part by NOAA, National Sea Grant College Program, Department of Commerce, under grant number NA85AA-D-SG140, project number R/F-115, through the California Sea Grant College Program. The U.S. Government is authorized to reproduce and distribute for governmental purposes. TABLE OF CONTENTS

Page

ABSTRACT ...... i i ACKNOWLEDGMENTS ...... v LIST OF FIGURES ...... x LIST OF TABLES ...... xiii INTRODUCTION ...... 1

MATERIALS AND METHODS ...... 6 Larval Specimens ...... 6 Culture Techniques ...... 7

Species Descriptions ...... 11 Comparisons of Cultured and Wild Larvae ...... 14 Comparisons of Cultured and Previously Described

Sebastes Larvae ...... 16 Morphometric Analyses ...... 18 RESULTS ...... 2 0 Pigment Patterns of Cultured Larvae ...... 20 Comparisons of Cultured and Wild Larvae ...... 23 Comparisons of Cultured and Previously Described

Sebastes Larvae ...... 25 Morphometries and Observations of Cultured Larvae ...... 28

Vll Vlll

Page

DISCUSSION ...... 3 0 Application of Morphometries in Identification of Larval Sebas tes...... 3 0 Application of Pigment Patterns in Identification of Larval Sebastes ...... 31 Evolutionary Significance of Larval Pigmentation ...... 33 Conclusions ...... 34 BIBLIOGRAPHY ...... 3 6 FIGURES ...... 4 5 TABLES ...... 6 0

APPENDIX A: An original Basic program to compare larval pigment patterns within a species ...... 77

APPEl~IX B: An original Basic program to compare larval pigment patterns between two species ...... 79

APPENDIX C: References for mystinus-type Sebastes compared in Figures 6-8 and Tables 6 & 7 ...... 81

APPENDIX D: References for chrysomelas-type Sebastes compared in Figures 9-11 and Tables 8 & 9 ...... 82

APPENDIX E: PSI comparisons of pigment patterns described for 1-2 day old cultured larvae of S. mvstinus and mystinus-type Sebastes Jmown to occur in Monterey Bay .... 83 IX

Page

APPENDIX F: PSI comparisons of pigment patterns described for cultured larvae of S. mystinus and mystinus-type Sebastes L4 days of age known to occur in the area of Monterey Bay ...... 84

APPENDIX G: PSI comparisons of pigment patterns described for cultured larvae 1-2 and L4 days of age of S. mystinus and mystinus-type Sebastes known to occur in the area of Monterey Bay ...... 8 5

APPENDIX H: PSI comparisons of pigment patterns described for 1-2 day old cultured larvae of S. chrysomelas and chrysomelas-type Sebastes known to occur in the area of Monterey Bay ...... 8 6

APPENDIX I: PSI comparisons of pigment patterns described for cultured larvae of S. chrysomelas and chrysomelas-type Sebastes L4 days of age known to occur in the area of Monterey Bay ...... 87

APPENDIX J: PSI comparisons of pigment patterns described for cultured larvae 1-2 and >4 days of age of S. chrysomelas and chrysomelas-type Sebastes known to occur in Monterey Bay ...... 8 8 LIST OF FIGURES

Figure Page

l. Collection site and study area ...... 45

2. Illustrations of cultured Sebastes mystinus larvae ..... 46

3. Illustrations of cultured Sebastes chrysomelas larvae ...... 4 7

4. Melanophore map of 27 pigment loci on a generalized Sebastes larva ...... 48

5. Melanophore map of 33 pigment loci on a generalized Sebastes larva ...... 49

6. Cluster diagram based on the similarities between pigment patterns of 1-2 day old cultured S. mystinus and previously described mystinus-type Sebastes larvae that occur in the area of Monterey Bay ...... 50

7. Cluster diagram based on the similarities between pigment patterns of preflexion cultured S. mystinus and previously described mystinus-type Sebastes larvae 24 days of age that occur in the area of Monterey Bay ...... 51

X XI

Figure Page

B. Cluster diagram based on the similarities between pigment patterns of 1-2 and 24 day old cultured S. mystinus and previously described mystinus-type Sebastes larvae that occur in the area of Monterey Bay ...... 52

9. Cluster diagram based on the similarities between pigment patterns of 1-2 day old cultured S. chrysomelas and previously described chrysomelas-type Sebastes larvae that occur in the area of Monterey Bay ...... 53

10. Cluster diagram based on the similarities between pigment patterns of preflexion cultured s. chrysomelas and previously described chrysomelas-type Sebastes larvae 24 days of age that occur in the area of Monterey Bay ...... 54

11. Cluster diagram based on the similarities between pigment patterns of 1-2 and 24 day old cultured S. chrysomelas and previously described chrysomelas­ type Sebastes larvae that occur in the area of Monterey Bay ...... 55 XII

Figure Page

12. Growth curves of notochord length and body depth at anus of cultured S. mystinus and S. chrysomelas larvae...... 56

13. Growth curves of snout to anus length and body depth at pectoral fin of cultured S. mystinus and S. chrysomelas larvae ...... 57

14. Growth curves of head length and pectoral fin length of cultured S. mystinus and S. chrysomelas larvae ...... 58

15. Morphometric proportions of 1-2 day old S. mystinus and S. chrysomelas larvae ...... 59 LIST OF TABLES

Table Page

1. Frequency of occurrence of melanophores at 33 pigment loci for S. mystinus larvae from four cultured broods and wild mystinus-type Sebastes larvae collected in 10 plankton tows ...... 6 0

2. Frequency of occurrence of melanophores at 33 pigment loci for S. chrysomelas larvae from three cultured broods and wild chrysomelas-type Sebastes larvae collected in 14 plankton tows ...... 62

3. Comparisons of larval pigment patterns within three cultured broods of S. mystinus and S. chrysomelas, wild mystinus-types collected in 10 plankton tows and chrysomelas-types collected in 14 plankton tows ..... 64

4. Comparisons of larval pigment patterns between different ages of cultured S. mystinus or S. chrysomelas and wild mystinus or chrysomelas-type larvae from plankton tows ...... 65

5. Comparisons of larval pigment patterns between same age cultured S. mystinus or S. chrysomelas and wild mystinus or chrysomelas-type larvae from plankton

tows ...... 66

Xlll XIV

Table Page

6 0 Pigment patterns of cultured So mystinus and previously described rnystinus-type Sebastes larvae 1-2 days of age that occur in the area of Monterey

Bay 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 o o o o o o o o o o o o o o o o o o o o 0 o o o o o o o o o o o o o o o o o 67

7 0 Pigment patterns of preflexion cultured So mystinus and previously described rnystinus-type Sebastes larvae L4 days of age that occur in the area of Monterey

Bay 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 o o o o o o o o o o o o o o o o 0 0 0 o o o 0 o 0 0 0 0 0 69

So Pigment patterns of cultured So chrysomelas and previously described chrysornelas-type Sebastes larvae 1-2 days of age that occur in the area of Monterey

Bay 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 o o o o o o o o o o o o o o o 0 0 o 0 o o o o o o o o 0 0 0 o o 0 0 71

9o Pigment patterns of preflexion cultured So chrysomelas and previously described chrysornelas-type Sebastes larvae L4 days of age that occur in the area of

Monterey Bay 0 0 0 0 0 0 0 0 0 0 0 0 0 0 o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o 7 3

10° Morphometries of cultured S. mystinus and

S. chrysomelas larvae 0 0 0 0 0 0 0 0 o o o o o o o o o 0 o o o o o o o o o o o o o • o o o 7 5

11° Morphological proportions of 1-2 day old cultured

So mystinus and S. chrysomelasoooooooooooooo••ooo.oooooo76 INTRODUCTION

The eastern north Pacific rockfishes (Sebastes, subfamily Sebastinae, family Scorpaenidae) are a speciose group of economically, recreationally and ecologically important fishes. Approximately 100 rockfish species occur worldwide, with the majority of species concentrating in temperate waters between 34° and 38° N latitude (Barsukov, 1982; Seeb, 1986). The North Pacific is the apparent center of origin and radiation for the genus Sebastes (Eschmeyer, 1971) with 72 species occurring in the eastern North Pacific (Seeb and Kendall, 1991). Within the genus, the adult fishes exhibit morphological similarities and wide ranges of intra­ and interspecific color variations (Chen, 1971). The Sebastes are viviparous fishes (Boehlert and Yruclavich, 1984). Most species reproduce once annually, but a few are known to have two reproductive seasons a year (Kendall and Lenarz, 1986). Fecundity increases proportionally with female size, and brood size ranges from 105 -106 larvae per extrusion period (Kendall and Lenarz, 1986; Love et al., 1989). At extrusion, the larvae are essentially transparent with patterns of dark melanophores. They have functional jaws and digestive tracts, residual yolksacs and are able to feed (Boehlert and Yoklavich, 1984) . During the next 2-3 months, the larvae undergo a series of

1 2

metamorphic changes as they develop through preflexion, flexion, postflexion, transformation and juvenile phases. Rockfish larvae <7 mm standard length numerically dominate the ichthyoplankton collected within the California Cooperative Oceanic Fisheries Investigations (CalCOFI) California Current sampling area (Ahlstrom et al., 1978). Larval rockfishes are the dominant component of ichthyoplankton collected in Monterey Bay during November­ June (Wallace, 1988) . Descriptions of Sebastes larvae are necessary to address questions concerning larval transport and recruitment, vital to ecological and fisheries applications. However, recent evolutionary divergence of the genus, large numbers of sister species (Barsukov, 1982; Seeb, 1986), and similarities in morphology and pigmentation have made larval species identifications problematic (Kendall, 1991) . Complete developmental series are available for 8 of the 72 species of eastern North Pacific Sebastes, with partial series available for another 53 species (Kendall and Lenarz, 1986; Kendall, 1989; Moreno, 1990). The majority of "extrusion" Sebastes larvae have been opportunistically obtained and described from intraovarian or prematurely-extruded larvae (Wales, 1952; Morris, 1955; Waldron, 1968; Efremenko and Lisovenko, 1970; Westrheim, 1975; Matarese et al., 1989; Kendall, 1991). Older specimens of field-collected Sebastes larvae have been used to 3

establish transformational series of postflexion through juvenile stages (Moser, 1971; Moser et al., 1977; Moser and Ahlstrom, 1978; Laroche and Richardson, 1979, 1981; Matarese et al., 1989) Less often, full-term larvae from known parental stock have been cultured to provide material for descriptions (Siokawa and Tsukahara, 1961; Moser and Butler, 1981, 1987; Stahl-Johnson, 1985; Canino and Francis, 1989; Kendall, 1989; Moreno, 1990) . To date, only a few individuals have survived in culture beyond preflexion. Prior to this study, there were no published descriptions of the larval stages of the black and yellow rockfish, S. chrysomelas, and only intraovarian larval descriptions of blue rockfish, S. mystinus (Wales, 1952; Efrernenko and Lisovenko, 1970). During the 1987-88 reproductive season, full-term larvae from field-collected females of these common, nearshore species were cultured at the Monterey Bay Aquarium in Monterey, California, as source specimens for descriptive analyses. Subsamples of the cultured la~-vae were used to develop descriptive criteria for e~trusion and preflexion S. mystinus and S. chrysomelas based on morphometries and pigmentation.

An expanded version of melanophore maps developed for larval Sebastes (Kendall and Lenarz, 1986; Moreno, 1990; Kendall, 1991) was used to describe pigment patterns. The 4

pigment patterns of cultured larvae were then compared to those of Monterey Bay Sebastes larvae from ichthyoplankton collections (Wallace, 1988) and to previously described larvae of species that occur in the Monterey Bay area (Moser et al., 1977; Stahl-Johnson, 1985; Kendall and Lenarz, 1986; Moser and Butler, 1987; Kendall, 1989; Matarese et al., 1989; Moreno, 1990) . Cluster analysis dendograms were generated from these comparisons and discussed in terms of allelic relationships shown in adult, juvenile and larval Sebastes (Seeb, 1986; Seeb and Kendall, 1991). The questions addressed were:

1. What is the intraspecific variability of pigment patterns and morphometric proportions

in these species of cultured Sebastes larvae?

2. Are the pigment patterns and morphometric

proportions demonstrated in cultured larvae similar to wild Sebastes larvae?

3. Are the pigment patterns or morphometric proportions of the majority of Sebastes larvae

useful for identification of field-collected specimens? 5

4. Can practical standards be established for description and identification of Sebastes larvae? MATERIALS AND METHODS

Larval Specimens

From mid-December 1987 to early January 1988, visibly gravid S. mystinus were collected at Stillwater Cove, Carmel Bay, California (Fig. 1). Field-caught females were taken by SCUBA divers using baited rockcod jigs suspended from weighted meter sticks. Gravid S. chrysornelas were collected later in the season (late February-early April 1988) from Stillwater Cove and display tanks at the Monterey Bay Aquarium (MBA) . Females captured from display tanks were taken by SCUBA divers using handnets. All females were transported to culture tanks at MBA, and held until larval extrusion occurred. In all cases, extrusions occurred during the night or early morning hours. For six of the eight fish that spawned, extrusions occurred as two or more pulses separated by 48-72 hours. The other two fish were preserved immediately following their first extrusions. Although batch extrusions have been observed by MBA husbandry personnel, their occurrence has not been noted in Sebastes literature.

Following extrusion, the parental females were removed from the culture tanks, sacrificed, preserved in 6 7

buffered 10% formalin solution and deposited as voucher specimens in the Moss Landing Marine Laboratories Ichthyology Collection. Newly-extruded larvae were stocked at densities of several thousand per tank, treated with a broad spectrum antibacterial, antifungal, antiparasitic medication (10% nifurpirinol "Prefuran") for three days and on a periodic basis thereafter. Viable S. mystinus larvae were periodically subsampled from the broods of three different females. The broods survived 9, 12 and 17 days. A fourth brood of

S. mystinus larvae were subsampled only at birth. An atypical subsample (n = 16) of 9 day old S. mystinus lacked pigments on the lower jaw, head, and nape. This was interpreted as a preservation problem and these larvae were not used in pigmentation analyses. The S. chrysomelas larvae were periodically subsampled from three broods. Their survivorship was 14, 21 and 23 days.

Culture Techniaues

Algal culture. Unicellular algae were cultured as food for marine rotifers (Brachionus plicatilis) . The seawater for algal cultures was sterilized in 20 liter nalgene carboys heated to boiling point with a 1000 watt 8

immersed titanium heating element. Seawater cooled to room temperature was enriched with a solution of major nutrients (formula "F-1": 1 ml/1), trace metals (1 ml/1) and vitamins (0.5 ml/1) (Guillard, 1975). Monoculture stock solutions of unicellular green algae (Tetraselmis, Dunaliella) and brown algae (Isochrysis, Monochrysis, Phaeodactylum) were obtained from MBA to inoculate the cultures. Heavily-aerated, room temperature cultures were lighted 24 hours a day with full­ spectrum bulbs. Rotifer cuJture. Marine rotifers (Brachionus plicatilis) were grown in static cultures as food for the larval rockfishes. Culture containers were 114 liter, plastic trashcans. Cultures were maintained in heavily­ aerated 27-32oc sterile seawater. The rotifers were fed a herring oil and egg yolk mixture (1:1) at the rate of 1.25 mg/1/day and Brewer's yeast at 1.5 mg/rotifer/day (Ohrun, MS in prep.). This diet was supplemented with unicellular algae according to availability. Once a day the air supply to the rotifer cultures was shut off, the container walls scrubbed with a stiff brush, and detritus siphoned from the bottoms. Three quarters of the culture volume was sieved through a 60 ~ screen retaining rotifers >60 ~ as food for the larval fishes. Fifty percent of the sieved seawater and smaller rotifers were returned to culture. The other quarter volume 9

of sieved seawater was discarded and replaced with an equal volume of sterile seawater or seawater containing unicellular algae. One quarter volume of each rotifer culture was left unharvested so that a stock of egg-bearing females was retained. Larval culture. Culture tanks were 400 liter cylinders with conical bottoms made of molded black plastic. The tank bottoms were painted white with an epoxy-resin paint so the black eyes of transparent larvae were visible.

Lighting was provided by overhead fluorescent bulbs and east­ facing windows. Water in the culture tanks was circulated from bottom to top to increase suspension of live food in the water column (M. Eldridge, NMFS Bodega pers. comm.). Incurrent water was piped through a valve in the bottom of the tank. Excurrent water was exchanged through portals cut in a center standpipe and screened with 330 !1 "Nitex" mesh. Ambient temperature seawater, supplied at low flow-through rates, was filtered to 7 !1 by MBA's sand filter system.

Final filtration to 1 !1 was provided by a polyester "Filtrex" cartridge.

Seawater from the culture tanks was tested for temperature, ammonia and pH on a weekly basis by MBA's research department. Temperatures ranged from 11.2-13.9°C, ammonia 0.29-2.19 pp/1000, and pH 8.03-8.12. 10

Larvae were fed cultured rotifers and wild zooplankton (Moser and Butler, 1981) Zooplankton were collected with a 300 ~ mesh plankton net towed at the seaward end of Elkhorn Slough, Moss Landing. Plankton were sieved through a 140 ~ screen to remove any larval competitors or predators. The 140 ~ fraction (e.g., polychaete larvae, barnacle nauplii, crab zoea, rotifers) was rinsed in sterile seawater and aerated with 1 g/100 1 of 10% nifurpirinol one hour before feeding. Live food was introduced to the tank twice a day. Flow-through seawater to the larval culture tanks was shut off for 4 hour periods during this time to maintain food concentrations. Fine particles of yeast from the rotifer cultures passed through the 60 ~ sieve and settled out in the larval culture tanks. Yeast that accumulated on the tank bottom became food for ciliate contaminants. Histology sections, prepared by the University of California Davis Veterinary Science School of Medicine, indicated that larval mortalities from the bottom of culture tanks were being penetrated and consumed by ciliates and bacteria. Histology sections of viable larvae collected from the water column indicated that those individuals were not subject to ciliate predation or bacterial contamination. Detritus and yeast was cleaned off the tank bottoms on a daily basis with disinfected sponges and a 5 mm diameter 1 1

siphon. The cleaning procedure reduced growth of a pink bacteria colonizing the bottom of the tanks (probably a Ps3udomonas bacteria, based on observations of plated colonies). Large numbers of the youngest larvae (<10 days old) were unable to avoid the siphon and were lost during the cleaning process. Larval escape response increased with age.

Species Descriptions

Larval fish from each brood were subsampled at birth and approximately every 4 days for the duration of the culture period. Since survivorship of the broods differed, the ages of subsamples varied. The subsampled larvae were refrigerated or anesthetized with tricaine methanesulfonate prior to preservation to prevent contortion of the specimens (B. MacCall, NMFS Tiburon, pers. comm.). Following Lavenberg et al. (1983), larvae were fixed in a 5% formalin and seawater solution buffered with sodium borate. Within a month of fixation the larvae were transferred to 25% isopropyl alcohol. A week later the isopropyl solution was increased to 50%. After another week, the larvae were transferred to 7 0% ethanol for final curation. During all stages of preservation and curation, specimens were stored in the dark to prevent the melanophores from fading. 1 2

Anesthetized or recently-fixed larvae were photographed through an Olympus dissecting microscope using 30X-64X magnification with 200 ASA Ektachrome film and transmitted light. Photographic slides of the larvae were projected through an enlarger to outline illustrations. Details of melanophores and pectoral fins were hand-drawn from magnified preserved specimens. Final drawings were done in pen and ink and watercolors (Figs. 2-3). Moser and Butler (1987) described three characteristic pigment patterns of larval Sebastes. The majority of species have one of two patterns: (1) a dorsal and ventral midline series from birth, or (2) only a ventral midline series. A third less common type has a ventral midline series at birth, with a rapidly developing dorsal midline series. Moser and Butler's pigmentation categories and patterns observed in the cultured larvae are used here to define mystinus-type and chrysomelas-type Sebastes larvae for comparisons between: (1) cultured larvae, (2) multi-species mixes of wild larvae and (3) larvae of species previously described in the literature. By this definition, mystinus­ type larvae have a short series of ventral midline melanophores, no melanophores on the dorsal midline, the Pectoral fins may or may not be pigmented (Table 1; Fig. 2). Chrysomelas-type larvae have a long series of ventral midline melanophores, a short series of melanophores on the dorsal 1 3

midline and unpigmented pectoral fins (Table 2; Fig. 3). Larvae of the third type are not considered here. One to two day old larvae of cultured S. mystinus and S. chrysomelas had notably less pigment than older preflexion larvae (Figs. 2-3). By 4 days of age, the pigment patterns of both species were essentially established, although the number of melanophores at a given locus increased throughout the preflexion period. Based on these observed differences in melanophore development, 1-2 day old larvae and larvae L4 days old were treated separately in pigmentation comparisons. Kendall and Lenarz (1986) developed a 26 loci map for describing the pigment patterns of larval Sebastes. As part of our Sea Grant research, Moreno and Wold expanded Kendall and Lenarz's map by the addition of 5 loci (Moreno, 1990). Since that time, Kendall (1991) has expanded his map to include a 27th locus in the dorsal midline series (Fig. 4). Here Kendall's 27 loci map was used to compare the pigment patterns of cultured and previously described Sebastes larvae, and a 33 locus map that combines Kendall's and Moreno and Wold's maps was used to compare the pigment patterns of cultured and wild larvae collected in plankton tows (Fig. 5). 1 4

Comparisons of Cultured and Wild Larvae

Wild preflexion larvae conforming to both pigment types were used in comparisons with cultured larvae. These were obtained from collections made in Monterey and Carmel

Bays, December 1985-March 1986, with a 248 ~ meter net or a

333 ~ bongo net (Wallace, 1988) . Larvae were preserved with ten percent buffered formalin solution. Mystinus-type larvae were selected from three tows made during March-April 1984 (n = 4) and seven tows late

November 1984 (n = 44) . Chrysomelas-type larvae were selected from six tows made during late March-early April

1984 (n = 23), three tows early June 1984 (n = 5), four tows late November 1984 (n = 12) and one tow early January 1985

(n = 8) . Selection of the first 48 larvae of either type was made without prior knowledge of tow dates. The presence or absence of melanophores at each pigment locus was compiled for individual cultured larvae 1-2 days of age and L4 days of age, and for wild preflexion mystinus and chrysomelas-type larvae of mixed species and unknown ages (Tables 1-2). Size was not a factor in selection of wild larvae. Percent Similarity Index (PSI) values of individual pigment patterns were derived from iterative comparisons of the pigment loci when: 1 5

PSI = B/A x 100, where,

no melanophores at locus n = 0;

melanophores at locus n = 1;

A= the total number of loci;

B = the number of matching values in the locus pairings; and

n = the number of iterations.

This index ranges from 0 with no similarity to 100 with no difference. Comparisons were made: (1) intraspecifically within the two age categories of cultured S. mystinus or S. chrysomelas; (2) intraspecifically between the two age categories of cultured S. mystinus or S. chrysomelas; (3) interspecifically between the two age categories of cultured S. mystinus and S. chrysomelas; (4) within field­ collected mystinus-type or chrysomelas-type larvae; chrysomelas-type larvae; and (6) between both species of cultured larvae and field-collected mystinus-type or chrysomelas-type larvae.

Comparjsons of Cultured and Previously Described Larvae

Kendall and Lenarz (1986) compiled the pigment patterns of 48 species of previously described, preflexion Sebastes larvae. The descriptions came from a variety of 1 6

sources and included intraovarian, prematurely-extruded, full-term and yolk-exhausted specimens. Among the species described, 38 have distributions off central California (Miller and Lea, 1972; Eschmeyer et al., 1983). Twenty of these have mystinus-type or chrysomelas-type pigment patterns and are likely to occur in the Monterey Bay area (Kukowski, 1972). Pigment patterns of these species plus 5 other species whose larvae were recently described [S. flavidus (Kendall, 1989), S. melanops (Kendall, 1989; Moreno, 1990), S. rastrelliger, S. atrovirens, S. carnatus (Moreno, 1990)] were compared to those of S. mystinus and S. chrysomelas larvae using Kendall's 27 loci map. The pigment patterns and ages of these species were verified by referring to the original or published illustrations referenced in Appendices C-D. When illustrations were not available, the pigment patterns compiled by Kendall and Lenarz (1986) were used for comparison. In either case, pigment patterns were assumed to be representative of the dominant condition of a species at a specified developmental stage. Pigments that occurred in L50% of S. mystinus or S. chrysomelas larvae were considered representative for each species. Comparisons of pigment patterns were made using the IBM program Statpro. The program could not compare more than 24 loci, so 3 loci that did not demonstrate intraspecific 1 7

variability were omitted from computations. The program used an iterative process to compare the pigment patterns of S. mystinus, S. chlcvsomelas, mystinus-type and chrysomelas­ type larvae. The output is displayed as clusters indicating the relative distance, or percent similarity, between pigment patterns (Figs. 6-ll; Appendices E-J). A decreasing relative distance between clusters is indicative of increasing similarity between species or developmental stage (e.g.,

0 = no difference between pigment patterns) when:

A + B/A + B + C = mean distance between clusters, where,

A = pigment present matches;

B = pigment absent matches;

C = the non-matches at each locus; and

l - mean distance = percent similarity

Relative distance values and species clusters were used to generate comparative dendograms of: (l) l-2 day old S. mystinus, intraovarian and full-term mystinus-type larvae of previously described species, (2) S. mystinus and Preflexion larvae L4 days of age of previously described mystinus-type species, (3) a combination of S. mystinus and mystinus-type larvae in groups l and 3, (4) l day old 1 8

S. chrysomelas larvae and larvae of previously described chrysomelas-type species, (5) S. chrysomelas larvae and preflexion larvae L4 days of age of previously described chrysomelas-type species and (6) a combination of S. chrysomelas and chrysornelas- type larvae in groups 4 and 5, all from the Monterey Bay area. Pigmentation similarities are discussed in terms of allelic relationships demonstrated in adult, juvenile and larval Sebastes (Seeb, 1986; Seeb and Kendall, 1991).

Morphometric Analyses

Morphometries, as delineated by Richardson and Laroche (1978) and Fahay (1983), were taken from recently preserved larvae using an Olympus CUE-II Image Analyzer. In this system, a video camera photographed larval fish through a dissecting microscope and projected the images on a television screen. When the images were analyzed, the computer program adjusted for changes in magnification and allowed user selection of units of measure. Larval dimensions were measured to the nearest 0.1 rnrn and include notochord length (NL) , total length (TL) , head length (HL) , eye diameter (ED) , snout length (SNL) , snout to anus length (SAL), body depth at anus (DA), body depth at pectoral fin 1 9

(DPF) , and pectoral fin length (PFL) . Growth curves of NL, HL, SAL, DA, DPF, and PFL were plotted.

Culture success affected the scope of morphological comparisons. Subsamples of progressively older fish decreased to as few as two individuals from a single brood. In addition, yolk-exhausted S. mystinus larvae exhibited morphological indicators of starvation such as shrinking, soft tissue collapse, anteriorly protruding angle of the pectoral girdles, "duck-billed" jaws and disproportionately large heads (Theilacker, 1978; Blaxter, 1988). Because of these malformations, the morphometric proportions of older S. mystinus larvae from the first and second broods were considered abnormal and were not used in comparisons. The development of older S. mystinus larvae from the third brood were considered normal and used for morphometric comparison with S. chrysamelas. Mean measurements of SAL, HL, DPF, DA and PFL were plotted as proportions of notochord length for each species. Proportions were arcsine transformed and significant differences calculated using Student's t-test. RESULTS

Pigment Patterns of Cultured Larvae

Sebastes mystinus and S. chrysomelas exhibited distinctively different pigment patterns. Both age categories of S. mystinus were less pigmented than s. chrysomelas larvae. Most notable were the dorsal midline series and longer ventral midline melanophore series of

S. chrysomelas and the pigmented fins of older S. mystinus

(Tables 1-2). In both species, larvae 1-2 days old had less pigment than older larvae, but melanistic pigments increased rapidly with age (Figs. 2-3). By 4 days of age, the pigmentation patterns were essentially established for the duration of the culture period.

Sebastes mystinus: 1-2 day old larvae. Pigments of young S. mystinus consisted of a short row of small, single stellate melanophores on the ventral midline (loci 23-24) and large, dark stellate melanophores formed a melanistic saddle over the stomach and digestive tract (loci 11, 17) (Figs. 2,

5; Table 1). These early larvae lacked pigments on the mandibular symphysis (locus 1), cranium (loci 4-5) and nape (locus 6) .

20 21

Sebastes mystinus: 4-17 daY old larvae. In addition to the pigments listed above, the majority of older larvae (>60%) had melanophores on the mandibular symphysis (locus 1), posterior margin of the pectoral fin (locus 9), and an internal cleithral pigment spot visible through the transparent body wall (locus 28) (Fig. 2, 5; Table 1). Less than 50% of larvae in this age class had pigments at other locations (Table 1) . Sebastes chrysomelas: 1 dav old lat-vae. At birth S. chrysomelas larvae had a complete ventral midline series

(loci 22-24) that was contiguous with the digestive tract pigment (loci 11, 17), a short dorsal midline series of large dendritic melanophores (loci 19-20), a faint smudge of pigment on the ventral surface of the stomach (locus 10), and melanophores on the anus (locus 16) (Figs. 3, 5; Table 2). A gap of one or more myomeres often separated melanophores at loci 19-20 (Fig. 3). Melanophores on the dorsum of the digestive tracts and stomachs (loci 11, 17) were small, indistinct and appeared as a smudged line. Sebastes chrysomelas: 4-23 day old larvae. The majority of older S. chrysomelas larvae (80-100%) had melanophores on the hindgut (locus 16), nape (locus 6), cranium (locus 4), cleithrum (locus 29), mandibular symphysis (locus 1) and behind the opercular openings (locus 30) 22

(Figs. 3, 5; Table 2). Melanophores at other loci occurred in less than 50% of the larvae and were not addressed here (Table 2).

Comparisons of Cultured and Wild Larvae

Comparisons within age categories and wild types. The pigment patterns of 1-2 day old S. mystinus from three different brood stocks had a PSI value of 78 (Table 3). Older S. mystinus larvae demonstrated increasing variability in pigmentation patterns and a corresponding decrease of PSI value (57) as melanophores developed on the cranium, nape, pectoral fins, and cleithrurn or accrued on the stomach and digestive tract (Table 3) . The PSI value of 1 day old S. chrysomelas larvae from three different brood stocks was 71 (Table 3). The pigment patterns of these larvae showed decreasing variability with age and a corresponding increase in PSI value (86) (Table 3) . The PSI values of field-collected mystinus-types and chrysomelas-types were 82 and 86, respectively (Table 3). Comparisons between age categories and wild types. PSI values from intraspecific comparisons between the two age categories of cultured s. mystinus and S. chrysomelas larvae were 65 (Table 4) . Both age categories of S. mystinus larvae 23

and wild mystinus-types showed similar variability (PSI 68 vs. 73). One day old S. chrysomelas are were more similar to wild chrysomelas-types than were S. chrysomelas 24 days old (PSI 89 vs. 70). Comparisons between cultured species or wild types. The PSI value from interspecific comparisons between pigment patterns of 1-2 day old cultured S. mystinus and S. chrysomelas (80) were slightly higher than those from intraspecific comparisons of this age class (78 and 71, respectively) (Tables 3, 5). As their melanophores developed, the pigment patterns of cultured S. mystinus and S. chrysomelas became increasingly dissimilar (64).

PSI values from comparisons between: (1) wild mystinus-types and chrysomelas-types, (2) 1-2 day old cultured S. mystinus and wild chrysomelas-types and (3) 1 day old cultured S. chrysomelas and wild mystinus-types were similar (PSI 75-76) (Table 5). Lower PSI values for comparisons of S. mystinus 24 days of age and wild chrysomelas-types (62) and S. chrysomelas 24 days of age and wild mystinus-types (68) reflected age-related melanophore development. 24

Comparisons of Cultured and Previously Described Larvae

Based on literature descriptions there were 16 Sebastes species, excluding S. mystinus, with mystinus-type and 11 species with chrysomelas-type larvae which could co-occur with S. mystinus and S. chrysomelas in the Monterey Bay area (Appendices C-D) . Iterative comparisons of pigment patterns produced clusters of PSI values for cultured S. mystinus and previously described mystinus-types and cultured S. chrysomelas and previously described chrysomelas­ types (Figs. 6-11; Appendices E-J). Within both mystinus­ types and chrysomelas-types similarities between pigment patterns appeared to be more reflective of developmental stage than species. Mystinus-type species complex. The mystinus-type complex included species S. serriceps, S. nigrocinctus, S. aleutianus, S. proriger, S. umbrosus and S. brevispinis that were either uncommon or rare in the Monterey Bay area. These species were not known to have periods of larval release coincident with S. mystinus in central California (Kendall and Lenarz, 1986; Echeverria, 1987). Two common species, S. constellatus and S. ruberrimus, (Miller and Lea, 1972; Hart, 1973) had much later periods of larval release (May-June versus November-March; Kendall and Lenarz, 1986; Echeverria, 1987) than s. mystinus. 25

Therefore, there were 8 common mystinus-type species - S. levis, S. paucispinis, S. miniatus, S. flavidus, S. diploproa, S. melanops, S. goodei and S. entomelas, (Miller and Lea, 1972; Eschmeyer et al., 1983) - that had extrusion periods coincident with S. mystinus in central California (Echeverria, 1987; Lea et al., in prep.). Of these, S. levis and S. paucispinis were readily distinguished from other mystinus-type species by their exceptionally large pectoral fins (Moser et al., 1977). Sebastes flavidus and S. melanops differed from S. mystinus 24 days of age because they laclced pigmentation on the pectoral fins (Kendall, 1989) . Serial descriptions were not available for S. diploproa and S. miniatus, but based on illustrations of later staged larvae (Moser et al., 1977; Matarese et al., 1989) it could be assumed the pectoral fins of preflexion larvae would be unpigmented. Larvae of the mystinus­ entomelas-goodei complex could not be distinguished using pigmentation (Fig. 8-9; Tables 6-7), reproductive timing or geographic range criteria.

Chrysornelas-tYQe species COIDQlex. Three of the 11 chrysomelas-type species - S. maliger, S. babcocki and S. semicinctus - were at the extremes of their ranges, and were rare or uncommon in the area (Miller and Lea, 1972; Eschmeyer et al., 1983; Hart, 1973). A fourth species, 26

S. dallii, was expected but unrecorded in the area (Kukowski, 1972) . None of these species were known to have periods of larval release coincident with S. chrysomelas in central California (Hart, 1973; Echeverria, 1987). Six common chrysomelas-type species - S. car.natus, S. atrovirens, S. auriculatus, S. caurinus, s. rastrelliger and S. saxicola - and one uncommon species - S. nebulosus - (Miller and Lea, 1972; Eschmeyer et al., 1983) had November­ May parturition periods coincident with S. chrysomelas (Echeverria, 1987; Moreno, 1990; Lea et al., in prep.; pers. obs.). Cultured larvae of S. caurinus, S. auriculatus (Stahl-Johnson, 1984) and S. nebulosus (Kendall, 1989) obtained from Washington stocks were more heavily pigmented in all early stages than other described species (Figs. 9-11; Tables 8-9). In contrast, young S. saxicola larvae had less pigmentation than other chrysomelas-type species and an exceptionally short dorsal midline series (Moser et al., 1977) (Fig. 8; Table 10). One to two day old larvae of cultured S. car.natus, S. atrovirens and S. rastrelliger (Moreno, 1990) were very similar and not distinguishable from S. chrysomelas (Fig. 9; Table 8; Appendix G). Pigment patterns developing in later stages or differences in extrusion periods may aid in distinguishing larvae of the chrysomelas-carnatus­ rastrelliger-atrovirens complex. For example, older 27

S. carnatus developed melanophores on the pectoral fin, S. rastrelliger was the only described chrysomelas-type with melanophores on the nares (Moreno, 1990) and S. atrovirens had a later season of parturition in central California (April-June) than the rastrelliger-carnatus-chrysomelas complex (late February-early April; Moreno, 1990; Lea et al., in prep.).

Morphometries and Observations of Cultured Larvae

Culture of S. mystinus and S. chrysomelas broods met with varying success. Larval die-offs of both species were heaviest during the first week. Subsequent survivorship and subsample sizes decreased over time (Figs. 12-14). At one week of age, larvae were feeding aggressively, both benthically and in the water column. They curled up laterally, in an anterior to posterior fashion, when perturbed and exhibited vigorous escape responses. Sebastes mystinus. The feeding regimes of different broods of larvae were dependent on availability of live food (i.e., cultured Brachionus plicatilis and wild zooplanlcton) and varied during the culture periods. Sebastes ~stinus larvae were fed a commercial lipid-protein formula (Kordon Corporation Artemia booster) with very low density rotifers 28

(<1/ml) because of a ciliate contamination of the Brachionus cultures at the time of extrusion. Bimodality was evident in the morphometries of 17 day old S. mystinus larvae. Some individuals were long and slender (e.g., 4.10 mm NL x 0.38 mm DA) and others were short and stout (e.g., 2.24 mm I~ x 0.63 mm DA). This is indicated by large standard deviations in the growth curves (Fig. 12). In addition, the yolk-exhausted S. mystinus larvae exhibited morphological indicators of starvation such as shrinking, soft tissue collapse, an anteriorly protruding angle to the pectoral girdle, "duck-billed" jaws and disproportionately large heads (Theilacker, 1978; Blaxter, 19 88) .

Sebastes chrysomelas. The first brood of S. chrysomelas larvae survived 14 days on the lipid­ protein/low rotifer diet. The mortalities were preserved for histology sections done at the University of California Davis Veterinary Science School of Medicine. Starvation was the probable cause of death since the alimentary tracts were devoid of digested material and 12 day old larvae from this brood demonstrated depressed growth (Figs. 12-14). The latter two broods of S. chrysomelas had improved survivorship and presumably normal growth on a combined diet of cultured rotifers and wild zooplankton (4-7 zooplankters/ml; Figs. 12-14). The larvae from the second 29

brood were terminated on the 15th day. The third brood was terminated on the 23rd day. ~~1 larvae were preflexion except for a single individual that underwent flexion prior to the 21st day. DISCUSSION

Application of Morphometries in Identification of Larval

Sebastes

With a few exceptions, preflexion Sebastes larvae lack distinguishing morphometric characters and are so similar that morphometric proportions are of limited value in species identification (Westrheim, 1975; Stahl-Johnson, 1985; Kendall and Lenarz, 1986; Moser and Butler, 1987; Kendall, 1989, 1991; Moreno, 1990; Seeb and Kendall, 1991; this study) . In addition, starvation was shown to affect the morphology of cultured larvae and can be a factor in the growth of wild larvae that are dependent on patchy distributions of zooplankton (Theilacker, 1978; Blaxter, 1988) . Clinal trends may also affect the morphometric comparisons of larval Sebastes. For example, full-term S. mystinus larvae collected in California at 36•N, 122•w averaged 3.8 mm TL (Wold, 1991) and eyed S. mystinus larvae collected in utero from Alaska at 59•N, 140•w averaged 5.2 mm TL (Efremenko and Lisovenko, 1970).

30 3 1

Application of Pigment Patterns in Identification of Larval Sebastes

As in the color-types of adult Sebastes, larval pigment patterns of closely related species are very similar. The majority of species have one of two larval patterns: (1) a dorsal and ventral midline series from birth or (2) only a ventral midline series (Moser and Butler, 1987). A third less common type has a ventral midline series at birth with a rapidly developing dorsal midline series. Sebastes mystinus, S. chrysomelas, 16 species of mystinus­ type and 11 species of chrysomelas-type larvae represent the two common patterns in the area of Monterey Bay. The culture of eastern North Pacific Sebastes larvae has met with limited success and few cultured larvae have been available for descriptive work (Moser and Butler, 1981, 1987; Stahl-Johnson, 1985; Canino and Francis, 1989; Kendall, 1989; Moreno, 1990). In this study and others (Kendall, 1989; Moreno, 1990) high intraspecific variability was demonstrated in the pigment patterns of cultured Sebastes larvae. The possibility that pigment patterns of cultured larvae were artifacts of culture and not representative of patterns found in nature had not been tested prior to this study. Pre-determined selection criteria based on observations of cultured S. mystinus and S. chrysomelas 32

larvae were used to select mystinus-type and chrysomelas-type larvae. As a result, the multi-species complexes of field­ collected larvae had higher similarity values than occurred in the monotypic cultures. However, the pigment patterns of cultured larvae and field-collected Sebastes larvae were similar. In most cases, the pigment patterns of same-age larvae of different species were more similar than those of the same species at different ages. Because pigments accrue rapidly after birth and Sebastes larvae metamorphose slowly it seemed probable that the pigment patterns of larvae L4 days of age would be represented in plankton collections.

However, similarities between 1-2 day old S. mystinus, S. chrysomelas and the wild larvae indicated the field collections were dominated by dense swarms of recently­ released, slow-moving larvae. Lack of a standard format to describe Sebastes larvae has undoubtedly lead to confusion when species identifications are made on the basis of literary descriptions. For example, illustrations of intraovarian (Wales, 1952) and prematurely extruded (Efremenko and Lisovenlco, 1970) S. mystinus represent a premature pigment pattern and do not include melanophores on the stomach and hindgut. In another example, the growth curves of 33

S. mystinus, S. rastrelliger, S. carnatus and S. atrovirens in Moreno (1990) demonstrate a decrease in standard length as a response to starvation or small sample size. Larval pigment patterns are apparently unaffected by condition factor although it should be noted that patterns presented as the smallest size range in Moreno's tables do not necessarily represent the youngest larvae. Dissimilarities in descriptions of the same species can also result because of clinal variation and limited parental stoclcs (Chen, 1986; Marliave, 1988; Matarese et al., 1989; Kendall, 1991). Sebastes mystinus larvae described here were very similar to Monterey Bay stocks of S. mystinus raised by Moreno (1990) but apparent differences in pigment patterns have been reported for California and Washington stocks of S. caurinus larvae (Moser et al., 1977; Stahl­ Johnson, 1985) and genetic stocks of S. caurinus (Stahl­ Johnson, 1985; Kendall, 1989).

Evolutionary Significance of Larval Pigmentation

Diagnostic allozyme loci have uniquely identified all but the most closely related species of adult, juvenile and larval Sebastes from known stocks (Seeb, 1986; Seeb and Kendall, 1991). However, genetic distances between some sister species such as S. chrysomelas and S. carnatus (0.040) 34

are similar to distances within a single species and need further discrimination (Seeb and Kendall, 1991). Seeb and Kendall (1991) collected larvae and selected adult controls based on coincident geographic ranges and periods of larval release to test the application of allozyme electrophoresis for identification of unknown species of co-occurring larvae. Not surprisingly, larvae that were sorted by identical pigment patterns represented closely-related, multi-species complexes. Not all the species represented in this study were analyzed by Seeb (1986), however similar relationship patterns were apparent when complexes of mystinus-type and chrysomelas-type larvae were compared to Seeb's genetic analyses of adult Sebastes. Relative to mystinus-types, the chrysomelas-types formed particularly tight genetic and color complexes. In both cases, the color complexes of mystinus­ entomelas-goodei and chrysomelas-carnatus were composed of closely related or sister species (Seeb, 1986) . Therefore, larval pigment patterns may represent evolutionary divergence and speciation of the Sebastes genus.

Conclusions

To make accurate interspecific comparisons it is essential that a standardized format be developed for the 35

description of Sebastes larvae. I suggest a format that: (1) follows a loci mapping process (Kendall and Lenarz, 1986; Moreno, 1990; Kendall, 1991) and (2) cites geographic area of collection, sample source, larval age and stage of development when available, frequency of melanophore occurrence, number of larvae examined and number of broods sampled. Sorting field-collected larvae on the basis of characteristic pigment patterns does not guarantee a monotypic cluster, but may be used to isolate small complexes of similar species with overlapping geographic ranges and coincident periods of larval release for further genetic analysis. If it can be assumed that environmental influences on larval survivorship such as upwelling, offshore advection and predation have a non-selective effect on Sebastes species represented in the plankton, cluster analysis will provide valuable information on larval dispersal and recruitment for ecological and fisheries management applications. BIBLIOGRAPHY

Anderson, K. L. 1979. A karyological investigation of the systematics of Sebastes (subgenus Pteropodus) . San Jose State University, unpublished Master of Arts thesis. 32 p.

Anderson, T. W. 1983. Identification and development of nearshore juvenile rockfishes (genus Sebastes) in central California kelp forests. unpublished Master of Arts thesis, California State University Fresno. 216 p.

Andrew, N. L. and B. D. Mapstone. 1987. Sampling and the description of spatial pattern in marine ecology. pp. 39-90. In M. Barnes (ed.), Oceanogr. Mar. Biol. Annu. Rev. 25. Aberdeen University Press.

Appelbaum, S. 1985. Rearing of the dover sole, Solea solea (L) , through its larval stages using artificial diets. Aquaculture 49: 209-221.

Arthur, D. K. 1976. Food and feeding of larvae of three fishes occurring in the California Current, Sardinops sagax, Engraulis mordax and Trachurus symmetricus. Fish. Bull. 74(3): 517-530.

Barrett, I., J. Joseph and G. Moser. 1966. Electrophoretic analysis of hemoglobins of California rockfish (genus Sebastodes). Copeia, No. 3: 489-494.

Barsukov, V. V. 1981. A brief review of the subfamily Sebastinae. J. Ichthyol. 21: 3-27.

Blaxter, J. H. S. 1988. Pattern and variety in development. pp. 1-58. In W. S. Hoar and D. J.Randall (eds.), Fish Physiology. Vol. XI, Part A: eggs and larvae. Academic Press, Inc.

Boehlert, G. W. 1977. Timing of the surface-to-benthic migration in juvenile rockfish, Sebastes diplopora, off Southern California. Fish. Bull. 75: 887-890.

Boehlert, G. W. 1981. The effects of photoperiod and temperature on laboratory growth of juvenile Sebastes diplopora and a comparison with growth in the field.

36 37

Fish. Bull. 79 (4): 789-794.

Boehlert, G. w. and M. M. Yoklavich. 1983. Effects of temperature, ration, and fish size on growth of juvenile black rockfish, Sebastes melanops. Env. Biol. Fish. 8 (1): 17-28.

Boehlert, G. W. and M. M. Yoklavich. 1984. Reproduction, embryonic energetics and the maternal-fetal relationship in the viviparous genus Sebastes (Pisces: Scorpaenidae). Biol. Bull. 167: 354-370.

Brownell, C. L. and D. A. Hortsman. 1987. Two low­ maintenance culture systems for small pelagic animals. Aquaculture 65: 375-383.

Buckley, L. J., T. A. Halavik, A. S. Smigielski and G. C. Laurence. 1987. Growth and survival of the larvae of three species of temperate marine fishes reared at discrete prey densities. pp. 82-92. In R. D. Hoyt (ed.), lOth Annual larval fish conference. Am. Fish. Soc. Symp. No. 2.

Cailliet, G. M., M. S. Love and A. W. Ebeling. 1986. Fishes: a field and laboratory manual on their structure, identification and natural history.pp. 127-128. Wadsworth Publishing Company, Belmont, CA.

Canino, M. and R. C. Francis. 1989. Rearing Sebastes larvae (Scorpaenidae) in static culture. Univ. WA FRI Tech. Rep. No. 8917.

Carlson, H. R. and R. R. Straty. 1981. Habitat and nursery grounds of Pacific rockfish, Sebastes spp., in rocky coastal areas of Southeastern Alaska. Mar. Fish. Rev. 43 (7): 13-19.

Charles, P. M., S. M. Sebastian, M. C. V. Raj and M. P. Marian. 1984. Effect of feeding frequency on growth and food conversion of cyprinus carpio fry. Aquaculture 40: 293-300.

Chen, Lo-chai. 1975. The rockfishes, genus Sebastes (Scorpaenidae), of the Gulf of California, including three new species, with a discussion of their origin. Proceedings of the calif. Acad. Sci., Fourth Series, 3 8

Vol. XL, No. 6. pp. 109-141.

Chen, Lo-chai. 1986. Meristic variation in Sebastes (Scorpaenidae), with an analysis of character association and bilateral pattern and their significance in species separation. U.S. Dep. Commer., NOAA Tech. Rep. NMFS No. 45: 1-17.

Cowgill, U. M., D. M. Williams and J. B. Esquivel. 1984. Effects of maternal nutrition on fat content and longevity of neonates of Dapbnia magna. J. Crust. Biol. 4 (2): 173-190.

Csengeri, I. and M. Petitjean. 1987. Fresh liver powder: a new starter diet for the larvae of a cyprinid fish. Aquaculture 65: 189-192.

Dabrowski, K. and R. Bardega. 1984. Mouth size and predicted food size preferences of larvae of three cyprinid fish species. Aquaculture 40: 41-46.

Dunn, J. R. 1983. Developmental osteology. pp. 48-50. In Ontogeny and systematics of fishes. Spec. Publ. No. 1, Am. Soc. Ichthyol. Herpetol.

Duray, M. and T. Bagarinao. 1984. Weaning of hatchery-bred milkfish larvae from live food to artificial diets. Aquaculture 41: 325-332.

Echeverria, T. W. 1987. Thirty-four species of California rockfishes: maturity and seasonality of reproduction. Fish. Bull. 85(2): 229-250.

Efremenko, V. N. and L. A. Lisovenko. 1970. Morphological features of intraovarian and pelagic larvae of some Sebastodes species inhabiting the Gulf of Alaska. pp. 267-286. In P. A. Moiseev (ed.), Soviet fisheries investigations in the northeast Pacific. Part V.

Eschmeyer, W. N., E. s. Herald and H. Hammann. 1983. A field guide to Pacific coast fishes of North America. pp. 132- 153. Houghton Mifflin Company Boston.

Fahay, M. P. 1983. Guide to the early stages of marine fishes occurring in the Western North Atlantic Ocean, Cape Hatteras to the Southern Scotian Shelf. J. N. W. 39

Atlan. Fish. Sci. 4. 423 p.

Fraser, A. J., J. R. Sargent, J. C. Gamble and P. MacLachlan. 1987. Lipid class and fatty acid composition as indicators of the nutritional condition of larval Atlantic herring. Am. Fish. Soc. Symp. 2: 129-143.

Gadomski, D. 1988. Reports from the Resources Enhancement and Hatchery Program (OREHAP), Redondo Beach, CA. In F. P. Binkowski (ed.), Early Life History Section Am. Fish. Soc. Newsletter 9(3): 7-8.

Green, R. H. 1971. Sampling design and statistical methods for environmental biologists. John Wiley & Sons, New York. 257 p.

Guillard, R. R. L., 1975. Culture of phytoplankton for feeding marine invertebrates. pp. 29-60. In W. L. Smith and M. H. Chanley (eds.), Culture of marine invertebrate animals. Plenum Press, New York and London.

Guillemot, P. J., R. J. Larson and W. H. Lenarz. 1984. Seasonal cycles of fat and gonad volume in five species of northern California rockfish (Scorpaenidae: Sebastes) . Fish. Bull. 83(3): 299-310.

Hallacher, L. E. 1974. The comparative morphology of extrinsic gasbladder musculature in the scorpionfish genus Sebastes (Pisces: Scorpaenidae). Proc. Calif. Acad. Sci., Fourth Series, Vol. XL(3): 59-86.

Hart, J. L. 1973. Pacific fishes of Canada. Fish. Res. Board Can. Bull. 180: 388-454.

Hurlbert, S. H. 1984. Pseudoreplication and the design of ecological field experiments. Ecol. Monogr. 54(2): 187- 211.

Kendall, A.W., Jr. 1991. Systematics and identification of larvae and juveniles of the genus Sebastes. Env. Biol. Fish. 30: 191-201.

Kendall, A.W., Jr. 1989. Additions to knowledge of Sebastes larvae through recent rearings. Northwest and Alaska Fisheries Center Processed Report 89-21. 46 p. 40

Kendall, A. w., Jr. and w. H. Lenarz. 1986. Status of early life history studies of northeast Pacific rockfishes. pp. 99-128. In Proceedings of the International Roclcfish Symposium, AK Sea Grant Rep. No. 87-2.

Kendall, A. w., Jr., E. H. Ahlstrom and H. G. Moser. 1983. Early life history stages of fishes and their characters. pp. 11-22. In Ontogeny and systematics of fishes. Spec. Publ. No. 1, Am. Soc. Ichthyol. Herpetol.

Kent, D. B. and R. F. Ford. 1985-86. Development of intensive culture technology, evaluation of juvenile population characteristics and habitat requirements, and assessment approaches to stocking for white seabass and California halibut. unpublished interim progress reports numbers 1-5.

Lam, T. J., J. V. Juario and J. Banno. 1985. Effect of thyroxine on growth and development in post-yolk-sac larvae of milkfish, Chanos ebanos. Aquaculture 46: 179- 184.

Laroche, W. A. and S. L. Richardson. 1980. Development of larvae and juveniles of the rockfishes Sebastes entomelas and S. zacentrus (family Scorpaenidae) and occurrence off Oregon, with notes on head spines of S. mystinus, S. flavidus, and S. melanops. Fish. Bull. 79(2): 231- 256.

Lavenberg, R. J., G. E. McGowen and R. E. Woodsum. 1983. Preservation and curation. pp. 57-59. In Ontogeny and systematics of fishes. Spec. Publ. No. 1, Am. Soc. Ichthyol. Herpetol.

Lenarz, w. and S. Moreland (eds.). 1985. Progress report on rockfish recruitment studies at the Tiburon Laboratory. Nat. Mar. Fish. Center Admin. Rep. 7-85-02. 19 p.

Love, M. S. and R. J. Larson. 1978. Geographic variation in the occurrence of tympanic spines and possible genetic differentiation in the kelp rockfish (Sebastes atrovirens). Copeia No. 1: 53-59.

Marliave, J. B. 1988. Variation in pigment and nape morphology of larval tidepool sculpin. pp. 80-88. In R. D. Hoyt (ed.), 11th Annual Larval Fish Conference. 4 1

Am. Fish. Soc. Symp. 5.

MacGregor, J. S. 1970. Fecundity, multiple spawning, and description of the gonads in Sebastodes. U.S. Dept. Int. u.s. Fish Wildlife Ser. Spec. Sci. Rep. Fish. No. 596. 12 p.

Matarese, A. C., A. W. Kendall, Jr., D. M. Blood, and B. M. Vinter. 1989. Laboratory Guide to Early Life History Stages of Northeast Pacific Fishes. U.S. Dep. Cammer., NOAA Tech. Rep. NMFS Circ. 80. 652 p.

Miller, D. J. and R. N. Lea. 1972. Guide to the coastal marine fishes of California. Calif. Fish Bull. No. 157: 90-113.

Moreno, G. 1990. Description of the larval stages of five northern California species of rockfishes (family Scorpaenidae) from rearing studies. unpublished Master of Science thesis, California State University Stanislaus. 68 p.

Morris, R. W. 1956. Early larvae of four species of rockfish, Sebastodes. Calif. Fish Game 42(2): 149-153.

Moser, H. G. 1967. Reproduction and development of Sebastes paucispinis and comparison with other rockfishes off southern California. Copeia No. 4: 773-797.

Moser, H. G. 1972. Development and geographic distribution of the rockfish, Sebastes macdonaldi (Eigenmann and Beeson, 1893), family Scorpaenidae, off Southern California and Baja California. Fish. Bull. 70(3): 941- 958.

Moser, H. G., E. H. Ahlstrom and E. M. Sandknop. 1977. Guide to the identification of scorpionfish larvae (family Scorpaenidae) in the eastern Pacific with comparative notes on species of Sebastes and Helicolenus from other oceans. NOAA Tech. Rep. NMFS Cir. No. 402. 71 p.

Moser, H. G. and E. H. Ahlstrom. 1978. Larvae and pelagic juveniles of blackgill rockfish, Sebastes melanostomas, taken in midwater trawls off Southern California and Baja California. J. Fish. Res. Board Can. 35: 981-996. 42

Moser, H. G. and J. L. Butler. 1981. Descriptions of reared larvae and early juveniles of the calico rockfish, Sebastes dallii. Calif. Coop. Oceanic Fish. Invest. Rep. 22: 88-95.

Moser, H. G. and J. L. Butler. 1987. Descriptions of reared larvae of six species of Sebastes. pp. 19-29. In W. H. Lenarz and D. R. Gunderson (eds.), Proceedings of the widow rockfish workshops. NOAA Tech. Rep., NMFS, U.S. Dept. Comm. No. 48.

Norton, J. G. 1986. Ocean climate influences on rockfish recruitment in the California Current. In Proceedings of the International Rockfish Symposium, AK Sea Grant Rep. No. 87-2.

Parrish, R. H., C. S. Nelson and A. Bakun. 1981. Transport mechanisms and reproductive success of fishes in the California Current. Biol. Oceanogr., 1(2}: 175-203.

Phillips, J. B. 1957. A review of the rockfishes of California. Fish Bull. No. 104: 1-158.

Rach, J. J. 1982. Water-stable diet for aquatic invertebrates. Prog. Fish-Cult. 44(2): 111-112.

Reguera, B. 1984. The effect of ciliate contamination in mass cultures of the rotifer, Brachionus plicatilis 0. F. Muller. Aquaculture 40: 103-108.

Richardson, S. L. and Laroche, W. A. 1979. Development and occurrence of larvae and juveniles of the rockfishes Sebastes crameri and Sebastes pinniger, and Sebastes helvomaculatus (family Scorpaenidae) off Oregon. Fish. Bull. 77(1): 1-41.

Seeb, L. W. and A. W. Kendall, Jr. 1991. Allozyme polymorphisms permit the identification of larval and juvenile rockfishes of the genus Sebastes. Env. Bio. Fish. 30: 191-201.

Seeb, L. W. and D. R. Gunderson. 1988. Genetic variation and population structure of the Pacific ocean perch (Sebastes alutus). Can. J. Fish. Aquat. Sci. 45: 78-88.

Seeb, L. W. 1986. Biochemical systematics and evolution of 43

the scorpaenid genus Sebastes. University of Washington, unpublished Ph.D. dissertation. 176 p.

Siokawa, T. and H. Tsukahara. 1961. Studies on habits of coastal fishes in the Amakusa Islands. Part I: Early life history of the purple rockfish, Sebastes pachycephalus pachycephalus Temmick et Schlegel. Records of Oceanographic Works in Japan, Spec. No. 5. pp. 123- 127.

Simpson, K. L., P. S. Schauer, C. R. Seidel and L. M. Richardson. Project Officer A. D. Beck. 1979. Nutritional requirements of marine larval and juvenile fish. Nat. Tech. Info. Ser. 41 p.

Singer, M.M. 1984. Food habits of juvenile rockfishes (Sebastes) in a central California kelp forest. Fish. Bull. 83 (4): 531-541.

Stahl-Johnson, K. L. 1985. Description characteristics of reared Sebastes caurinus and Sebastes auriculatus larvae. pp. 65-76. In A. W. Kendall and J. B. Marliave (eds.), Descriptions of early life history stages of selected fishes. Can. Tech. Rep. Fish. Aquat. Sci. 1359.

Szlaminska, M. and A. Przybyl. 1986. Feeding of carp (cyprinus carpio L.) larvae with an artificial dry food, living zooplankton and mixed food. Aquaculture 54: 77- 82.

Theilacker, G. H. and M. F. McMaster. 1971. Mass culture of the rotifer Brachionus plicatilis and its evaluation as a food for larval anchovies. Mar. Biol. 10: 183-188.

Theilacker, G. H. 1978. Effect of starvation on the histological and morphological characteristics of jack mackerel, Trachurus symmetricus, larvae. Fish. Bull. 78: 403-414.

Tsuyuki, H., E. Roberts, R. H. Lowes, W. Hadaway and J. Westrheim. 1968. Contribution of protein electrophoresis to rockfish (Scorpaenidae) systematics. J. Fish. Res. Board Can. 25(11): 2477-2501.

Underwood, A. J. 1981. Techniques of analysis of variance in experimental marine biology and ecology. pp. 513-605. 44

In M. Barnes (ed.), Oceanogr. Mar. Biol. Annu. Rev. 19. Aberdeen University Press.

Van Ballaer, E., F. Amat, F. Hontoria, P. Leger and P. Sorgeloos. 1985. Preliminary results on the nutritional evaluation of w3-highly unsaturated fatty acid-enriched Artemia nauplii for larvae of the seabass, Dicentrarchus labrax. Aquaculture 49: 223-229.

Verreth, J., V. Storch and H. Segner. 1987. A comparative study on the nutritional quality of decapsulated Artemia cysts, micro-encapsulated egg diet and enriched dry feeds for Clarias gariepinus Burchell larvae. Aquaculture 63: 269-282.

Waldron, K. D. 1968. Early larvae of the canary rockfish, Sebastodes pinniger. J. Fish. Res. Board Can. 25(4): 801-803.

Wales, J. H. 1952. Life history of blue rockfish, Sebastodes mystinus. Calif. Fish Game 38: 485-498.

Walford, J. and T. J. Lam. 1987. Effect of feeding with microcapsules on the content of essential fatty acids in live foods for the larvae of marine fishes. Aquaculture 61: 219-229.

Wallace, F. R. 1988. Inshore-offshore and seasonal patterns: larval fishes in Monterey Bay, 1983-1985. California State University Hayward, unpublished Master of Science thesis. 83 p.

Westrheim, S. J. 1975. Reproduction, maturation, and identification of larvae of some Sebastes (Scorpaenidae) species in the Northeast Pacific Ocean. J. Fish. Res. Board Can., 32(12): 2399-2410.

Zar, J. H. 1984. Biostatistical analysis second edition. Prentice-Hall Inc., Engelwood Cliffs, N.J. 718 p. 45

MONTEREY BAY

...... ·:· ... ..

.· . .·.. .0 • .. BAY AQUARIUM . . . . .

. . ·. .. . .

CARMEL BAY ......

.. .

Figure 1. Collection site and study area. 46

Sebastes mystinus

·~ \ZJ ·-g~,-·······.. :... ~~.:.::.~~~: ::_12::=

x=3.Bmm Yolk sac

··-·····- ._

17 Days i=3.Bmm Preflexion

Figure 2. Illustrations of cultured Sebastes mvstinus larvae. 47

Sebastes chrysomelas - ~~'"1'~;;~\'-"':_.,_~~~~:~_::=:~:-~_::·: __ ~::c ~- .. · '' 1Day x=4.4mm Yolksac

8 Days Preflexion x=4.0mm

...... - -~~--...:.:;..·-

15 Days x=4.7mm Preflexion

Flexion x=5.0mm

Figure 3. Illustrations of cultured Sebastes chrysornelas larvae. 4 6 1117 18 19 20 21

i

7 10 1213 14 1.615 22 23 24 25 26 27 ~ \ '•g 8 Figure 4. Melanophore map of 27 pigment loci on a generalized Sebastes· larva (Kendall and Lenarz, 1986; Kendall, 1991). Pectoral fin shown separately.

-1>- 00 21

33 1 23 31

12 13 14 16 15 32 22 28 23 24 25 26 27

~\ ····g \a

Figure 5. Melanophore map of 33 pigment loci on a generalized Sebastes larva (expanded from Kendall and Lenarz, 1986; Moreno, 1990; Kendall, 1991). Pectoral fin shown separately.

-!>. 'Ci 50

s. aleutianus**

S. proriger*

S. ruberrimus*

S. diploproa*

S. mystinus*

s. nigrocinctus*

S. brevispinis**

s. flavidus*

S. melanops**

S. levis*

s. paucispinis*

*full-term larvae **premature larvae

Figure 6. Cluster diagram based on the similarities between pigment patterns of 1-2 day old cultured S. mystinus and previously described mystinus-type Sebastes larvae that occur in Monterey Bay. 5 I

S. constellatus

S. mystinus

s. goodei

S. ruberrimus

s. entomelas

S. tunbrosus

S. serriceps

S. miniatus

S. melanops

S. flavidus

S. paucispinis

Figure 7. Cluster diagram based on the similarities between pigment patterns of cultured s. mystinus and previously described mystinus-type Sebastes larvae L4 days of age that occur in Monterey Bay. 52

s. aleutianus**

s. proriger* s. ruberrimus*

s. serriceps+ s. diploproa* s. mystinus* s. levis*

s. miniatus+ s. nigrocinctus* s. cons tell a tus+

s. mystinus+ s. goodei+

s. ruberrimus+

s. entomelas+

s. umbrosus+ s. melanops+

s. brevispinis** s. flavidus* s. flavidus+

S. melanops**

S. paucispinis*

S. paucispinis+ *full-term larvae **premature larvae +larvae L4 days of age Figure 8. Cluster diagram based on the similarities between pigment patterns of 1-2 and L4 day old cultured S. mystinus and previously described mystinus-type Sebastes larvae that occur in Monterey Bay. 53

S. atrovirens

S. auriculatus

S. caurinus

s. babcocki

S. rastrelliger

S. maliger

S. carnatus

S. chrysomelas

s. saxicola

Figure 9. Cluster diagram based on the similarities between Pigment patterns of 1-2 day old cultured s. chrysomelas and Previously described chrysomelas-type Sebastes larvae that occur in Monterey Bay. 54

s. atrovirens

s. auriculatus

S. caurinuSl

S. nebulosus

S. caurinus2

S. rastrelliger

S. carnatus

S. chrysomelas

s. dallii

S. semicinctus

Figure 10. Cluster diagram based on the similarities between pigment patterns of cultured s. chrysomelas and previously described chrysornelas-type Sebastes larvae L4 days of age that occur in Monterey Bay. 55

s. atrovirens*

s. auriculatus*

s. auriculatus+

s. caurinus1+

s. nebulosus+

s. caurinus2+

s. caurinus*

s. a trovirens+

s. chrysomelaS+

s. carnatus+

s. rastrelliger+ s. babcocki*

s. maliger*

s. carnatus*

s. ras

s. dallii+

s. chrysomelas*

s. semicinctus+

s. saxicola*

*1-2 day old larvae +larvae 2_4 days of age

Figure 11. Cluster diagram based on the similarities between pigment patterns of 1-2 and 2.4 day old cultured S. chrysomelas and previously described chrysomelas-type Sebastes larvae that occur in Monterey Bay. 56

7

(2) (6) 6 (2) ~ (2) (9) J-- .<:: (26) .w t:n 5 I >:! (4 GJ '\ 'I ..:I \CB I 'd,.. I ' 4 tr 0 .<:: u 0 .w 0 (32) z 3

( I 0)

2 0 5 10 15 20 25 1.4 (2)

1.2 ~

{!) 1.0 \ ~ '\ .w \ (2) ro 0.8 .<:: ! .w 0. (9) GJ 1I ~ 0.6 I .a I B. cb.Iyaamelaa {n) 0 II S. mystinua (n) Pl 0.4 il! mean.± S.D. (I 0)

0.2 0 5 10 15 20 25 Age in Days

Figure 12. Notochord length and body depth at anus versus age of cultured Sebastes larvae. 57

1.2 (2)

1.0 (6)

(2) O.B r1 ! (9) (26 \ 1 1 (44~ ,I \csJ/ 0.6 \

(32) ( 10)

0.4+-~-r~~~~~~~~~~~-r~~~-r-r, 0 5 10 15 20 25

1.6

(2) (2) I .4 i 1.2 .r:: .w 8' 1.0 Gl ..:1

O.B I S. ch.tyaomelas (n) I S. myatinua (n) 0.6 2 mean .± s .D. (32) ( I 0)

0.4+-~~~--~~~~-r~,-~~-r~,-.-~~~ 0 5 10 15 20 25 Age in Days

Figure 13. Body depth at pectoral fin and head length versus age of cultured Sebastes larvae. 58

0.8

0.7

0.6

0.5

0."1

(32) ( 10) 0.3

0.2 0 5 10 15 20 25

2.5 (2) (2)

(6)

2.0 (9) i (26) .s::: .w IJ1 1.5 l:i OJ ...:1 UJ ;:l ~ (32) .w' (I 0) 11!11 ;:l s. chzyeomelae (n} 0 l:i I S. myetinue (n) 0.5 til ~ mean.±. S.D.

0 5 10 15 20 25 Age in Days

Figure 14. Pectoral fin length and snout to anus length versus age of cultured Sebastes larvae. 59

0 Sebastes mystinus n = 60 r:zl Sebastes chrysomelas n ~ 47 * significantly different t(O.Ol)

40

30 Ql -~ :=ffi 20 Ql CL 1P

-.!:. -.!:. .<::.­- .!:. ,so, -0> .!:.-C> - e>t: C>t: C>t: Q)-" Q) " Q) Q)-" Q) --o --oQ)- --o (/) ~ ::0 0 -o~ t;::_g" ~ tijti "'Q).!:. 0 Ciiu ::c (.) ~ 0 • 0 o­ so • B0 -o gz z ~z (f) a. • .

Figure 15. Means and standard errors of morphometric proportions of 1-2 day old S. mystinus and S. chrysomelas larvae as percentages of notochord length. 60

Table l. Frequency of occurrence of melanophores at 33 pigment loci for S. mystinus larvae from four cultured broods and wild mystinus-type Sebastes larvae collected in 10 plankton tows. 6 1

cultured cultured wild Locus S. mystinus S. mystinus mytinus-type 1-2 days old 4-17 days old larvae

(n) ( 46) (109) ( 48) 1 0 100 48 2 0 0 0 3 0 4 0 4 0 46 54 5 0 34 2 6 0 37 48 7 0 1 15 8 0 77 13 9 0 86 15 10 26 62 21 11 100 100 100 12 24 67 50 13 63 68 6 14 54 62 35 15 61 88 38 16 67 96 88 17 100 100 100 18 0 1 0 19 0 0 0 20 0 1 2 21 0 0 0 22 0 0 0 23 100 100 100 24 100 100 100 25 0 0 0 26 2 6 29 27 11 24 23 28 0 1 2 29 0 61 15 30 2 49 2 31 0 0 0 32 0 0 0 33 0 0 0 62

Table 2. Frequency of occurrence of melanophores at 33 pigment loci for S. chrysomelas larvae from three cultured broods and wild chrysomelas-type Sebastes larvae collected in 14 plankton tows. 63

cultured cultured wild Locus s. chrysomelas s. chrysomelas chrysomelas- 1 day old 4-23 days old type larvae

(n) (31) (56) (48) 1 0 80 17 2 0 7 0 3 0 4 4 4 0 86 46 5 0 4 2 6 45 98 31 7 0 16 2 8 0 0 0 9 0 0 0 10 100 100 60 11 100 100 100 12 0 5 2 13 0 2 0 14 10 40 19 15 0 14 0 16 97 100 71 17 100 100 100 18 10 29 33 19 100 100 100 20 100 100 100 21 0 0 0 22 100 100 100 23 100 100 100 24 100 100 100 25 3 13 19 26 0 0 0 27 0 0 2 28 48 48 15 29 0 88 0 30 0 89 2 31 0 7 0 32 0 13 0 33 0 0 0 64

Comparisons # loci Sx sx2 s2 PSI compared

S. mystinus 1-2 days old 1,035 811 637 0.62 78 (n = 46)

s. mystinus 4-17 days old 5,886 3,353 1,936 0.33 57 (n = 109)

s. chzysomelas 1 day old 465 330 235 0.51 71 (n = 31)

s. chzysomelas 4-23 days old 1 '711 1,472 1,284 0. 7 5 86 (n = 56)

mystinus- types 1,128 924 765 0.68 82 (n = 48)

chrysomelas- types 1,176 1,014 880 0. 75 86 (n = 48)

Table 3. Comparisons of larval pigment patterns within three cultured broods of S. mystinus and S. chzysomelas, wild mystinus-types collected in 10 plankton tows and chrysomelas-types collected in 14 plankton tows. 65

Comparisons # loci Ix Ix2 s2 PSI compared s. mystinus 1-2 days old vs. S. mystinus 2,576 1,663 1,089 0.42 65 4-17 days old

S. mystinus 1-2 days old vs. 2,208 1,806 1,492 0.77 68 my s tinus-types

S. mystinus 4-17 days old vs. 5,232 3,825 2,824 0.53 73 mystinus-types s. chrysomelas l day old vs. S. chrysomelas 1 '736 1,395 1 '13 3 0.96 65 4-23 days old

S. chrysomelas 1 day old vs. 1,488 1,322 1,178 0.79 89 chrysomelas-types

S. chrysomelas 4-23 days old vs. 2,688 2,128 1,705 0.90 79 chrysomelas-types

Table 4. Comparisons of larval pigment patterns between different ages of cultured S. mystinus and S. chrysomelas larvae and wild mystinus-type and chrysomelas-type larvae. 66

Comparisons # loci Sx sx2 s2 PSI compared

s. mys tinus vs . s. chrysomelas 1,426 1,136 908 0.59 80 1-2 days old

s. mystinus vs. s. chrysomelas 6,104 3,936 2,566 0.42 64 4-23 days old

mystinus-types vs. 2,352 1, 760 1,331 0.90 75 chrysomelas-types

s. mystinus 1-2 days old vs. 2,208 1,679 1,285 0.88 76 chrysomelas-types

s. mystinus 4-17 days old vs. 5,232 3,256 2,051 0.40 62 chrysomelas-types

s. chrysomelas 1 day old vs. 1,488 1,131 869 0.62 76 mystinus-types

s. chzysomelas 4-23 days old vs. 2,688 1,840 1,274 0. 72 68 mystinus-types

Table 5. Comparisons of larval pigment patterns between same age cultured S. mystinus and S. chrysomelas and wild rnystinus-type and chrysornelas-type larvae. 67

Table 6. Pigment patterns of cultured S. mystinus and previously described mystinus-type Sebastes larvae 1-2 days of age that occur in the area of Monterey Bay, California. Species codes and references are listed in Appendix C. 68

Spp. aleut brev diplo flav levis mel a myst nigr pauc 1 prori rube

Code I 2 3 4 5 6 7 8 9 I 0 I I

1 0 0 0 0 I 0 0 I 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 4 0 0 0 0 1 0 0 0 0 0 0 5 0 0 0 0 1 0 0 0 0 0 0 6 0 0 0 0 1 0 0 1 0 0 0 7 0 0 1 0 0 0 0 0 0 0 0 8 0 0 0 0 1 0 0 0 0 0 0 9 0 0 0 0 1 0 0 0 1 0 0 1 0 0 0 0 1 0 1 0 1 0 0 0 1 1 1 1 1 1 1 1 I 1 1 1 1 1 2 1 1 0 0 0 0 0 0 0 0 0 1 3 1 0 1 1 0 0 1 1 0 0 1 14 1 0 0 0 0 1 1 1 0 1 1 15 1 1 1 0 0 0 1 1 1 1 0 1 6 1 1 1 1 1 1 1 1 1 1 1 1 7 1 1 1 1 1 1 1 1 1 1 1 1 8 0 0 0 0 0 0 0 0 0 0 0 1 9 0 0 0 0 0 0 0 0 0 0 0 20 0 0 0 0 0 0 0 0 0 0 0 2 1 1 0 0 0 0 0 0 0 0 1 0 22 0 0 0 0 0 0 0 0 0 0 0 23 1 1 1 1 1 1 1 1 1 1 1 24 1 1 1 1 1 1 1 1 1 1 1 25 0 0 0 0 0 0 0 0 0 0 0 26 1 1 1 1 0 1 0 0 0 1 1 27 1 0 0 0 0 0 0 0 0 1 1

0 = no melanophores at locus n 1 = melanophores at locus n 69

Table 7. Pigment patterns of preflexion cultured S. mystinus and previously described mystinus-type Sebastes larvae L4 days of age that occur in the area of Monterey Bay, California. Species codes and references are listed in Appendix C. 70

Spp. cons en to flav good mel a ffillll myst pauc rube sern umb

Code 1 2 1 3 14 1 5 1 6 I 7 1 8 1 9 20 2 1 22

1 1 1 0 0 1 1 1 0 1 0 1 2 1 0 0 0 0 0 0 0 0 0 0 3 1 0 0 0 0 0 0 0 0 0 0 4 0 0 0 1 0 1 0 1 1 1 0 5 0 0 0 0 0 1 0 0 0 0 0 6 0 1 0 1 1 1 0 0 1 1 0 7 0 0 0 1 1 0 0 0 1 0 0 8 1 1 0 1 0 0 1 0 1 0 1 9 1 1 0 1 0 0 1 1 1 0 1 10 1 1 1 0 0 0 1 0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 12 1 0 0 1 0 1 1 0 1 0 0 1 3 1 1 1 1 0 0 1 0 1 0 1 14 1 1 0 1 0 0 1 0 1 1 1 15 1 0 0 1 1 1 1 0 1 1 0 16 1 1 1 1 1 1 1 1 1 1 1 17 1 1 1 1 1 1 1 1 1 1 1 1 8 0 0 0 0 0 0 0 0 0 0 0 1 9 0 0 0 0 0 0 0 0 0 0 0 20 0 0 0 0 0 0 0 0 0 0 0 21 0 0 0 0 0 0 0 0 0 0 0 22 1 0 0 0 0 0 0 0 0 0 0 23 1 1 1 1 1 1 1 1 1 1 1 24 1 1 1 1 1 1 1 1 1 1 1 25 0 0 0 0 0 0 0 0 0 0 0 26 0 0 1 0 0 1 0 0 0 1 0 27 0 0 0 0 0 0 0 0 0 0 0

0 = no melanophores at locus n 1 = melanophores at locus n 7 1

Table 8. Pigment patterns of cultured S. chrysomelas and previously described chrysomelas-type Sebastes larvae 1-2 days of age that occur in the area of Monterey Bay, California. Species codes and references are listed in Appendix D. 72

Spp. atro a uri bab earn caur chry mali rast sax I

Code 1 2 3 4 5 6 7 8 9

1 0 1 0 0 1 0 0 0 0 2 0 0 0 0 0 0 0 0 0 3 0 1 0 0 0 0 0 0 0 4 0 1 0 0 1 0 0 1 0 5 0 1 0 0 0 0 0 0 0 6 0 1 1 1 1 0 I 1 0 7 0 0 0 0 0 0 0 0 0 8 0 0 0 0 0 0 0 0 0 9 0 0 0 0 0 0 0 0 0 1 0 1 1 1 1 1 1 1 1 0 1 1 1 1 1 1 1 1 1 1 1 1 2 0 0 0 0 0 0 1 0 0 1 3 1 1 1 0 1 0 1 0 0 1 4 1 0 1 0 0 0 1 0 0 1 5 0 0 0 0 0 0 0 0 0 1 6 0 1 1 1 1 1 0 0 0 1 7 1 1 1 1 1 1 0 1 1 1 8 0 1 0 0 1 0 1 1 0 1 9 1 1 1 1 1 1 1 1 0 20 1 1 1 1 1 1 1 1 1 21 0 0 0 0 0 0 0 0 0 22 1 1 1 1 1 1 1 1 1 23 1 1 1 1 1 1 1 1 1 24 1 1 1 1 1 1 1 1 1 25 0 1 0 0 0 0 0 0 0 26 0 0 1 0 0 0 0 0 0 27 0 0 1 0 0 0 0 0 0

0 = no melanophores at locus n 1 = me1anophores at locus n 73

Table 9. Pigment patterns of preflexion cultured S. chrysomelas and previously described chrysomelas-type Sebastes larvae L4 days of age that occur in the area of Monterey Bay, California. Species codes and references are listed in Appendix D. 74

Spp. atro aun earn caur caur chry dall nebu rast sem1 1 2

Code 1 0 1 1 1 2 1 3 14 1 5 1 6 1 7 1 8 1 9

1 0 1 1 1 1 1 0 1 0 0 2 0 1 0 1 0 0 0 1 0 0 3 0 1 0 1 0 0 0 0 0 0 4 1 1 1 1 1 1 1 1 1 0 5 1 1 0 1 1 0 0 1 0 0 6 1 1 1 1 1 1 1 1 1 0 7 0 1 0 1 0 0 0 1 1 0 8 0 0 0 0 0 0 0 0 0 0 9 0 0 0 0 0 0 0 0 0 0 1 0 1 1 1 1 1 1 0 1 1 0 1 1 1 1 1 1 1 1 1 1 1 1 1 2 0 0 0 1 0 0 0 1 0 0 1 3 1 1 0 1 1 0 0 0 0 0 1 4 0 0 0 0 1 0 0 1 0 0 1 5 0 0 0 0 0 0 0 0 0 0 1 6 1 1 1 1 1 1 0 1 1 0 1 7 1 1 1 1 1 1 1 1 1 1 1 8 0 1 0 1 0 0 0 1 1 0 1 9 1 1 1 1 1 1 1 1 1 1 20 1 1 1 1 1 1 1 1 1 1 2 1 0 0 0 0 0 0 0 0 0 0 22 1 1 1 1 1 1 1 1 1 1 23 1 1 1 1 1 1 1 1 1 1 24 1 1 1 1 1 1 1 1 1 1 25 0 1 0 0 0 0 1 0 1 0 26 0 1 0 0 0 0 0 0 0 0 27 0 0 0 0 0 0 0 0 0 0

0 = no me1anophores at locus n 1 = melanophores at locus n 75

Measurement (mm) S. m ystinus (n) S. chrysomelas (n)

Notochord Length extrusion 3.8 ± 0.02 (3.5-4.1)* (62) 4.4 ± 0.03 (3.6-4.7)* (48) pre flexion 3.6 ± 0.05 (2.2-4. 7) (89) 4.6 ± 0.10 (3.6-4.7) (54) flexion - (0) 5.6 (1) Total Length extrusion 4.0 ± 0.02 (3.6-4.4)* (62) 4.6 ± 0.03 (3.7-4.9)* (48) pre flexion 3.8 ± 0.05 (2.4-5.0) (89) 4.8 ± 0.10 (2.6-6.6) (54) flexion - (0) 6.0 (1) Snout-Anus Length extrusion 1.2 ± 0.01 (1.0-1.3)* (62) 1.5 ± O.Dl (1.4-1.7)* (48) pre flexion 1.1 ± 0.01 (0.6-1.3) (89) 1.6 ± 0.04 (0.9-2.3) (54) tlexion - (0) 2.3 (1) Head Length extrusion 0.7 ± 0.01 (0.6-0.8)* (62) 0.8 ± 0.01 (0.6-0.8)* (48) pre flexion 0.7± 0.01 (0.4-0.9) (89) 0.9 ± 0.02 (0.6-1.4) (54) flexion - (0) 1.4 (1) Eye Diameter extrusion 0.3 ± 0.00 (0.2-0.4) (62) 0.3 ± 0.00 (0.2-0.3) (48) preflexion 0.3 ± 0.00 (0.2-0.5) (89) 0.3 ± 0.01 (0.2-0.5) (54) flexion - (0) 0.5 (1) Snout Length extrusion 0.2 ± 0.00 (0.2-0.3) (62) 0.2 ± 0.00 (0.2-0.3) (48) pretlexion 0.2 ± 0.00 (0.1-0.3) (89) 0.2± 0.01 (0.2-0.4) (54) flexion - (0) 0.3 (1) Depth at Pectoral Fin extrusion 0.5 ± 0.01 (0.4-0.6)* (62) 0.6 ± 0.00 (0.5-0.6)* (48) pre flexion 0.5 ± 0.01 (0.4-0.8) (89) 0.6 ± 0.02 (0.4-1.0) (54) flexion - (0) 1.1 (1) Body Depth at Anus extrusion 0.4 ± 0.01 (0.4-0.5)* (62) 0.5 ± 0.01 (0.4-0.6)* (48) pretlexion 0.4 ± O.Ql (0.3-0.6) (89) 0.6 ± 0.02 (0.4-0.9) (54) tlexion - (0) 1.2 (1) Pectoral Fin Length extrusion 0.4 ± 0.01 (0.3-0.5) (62) 0.4 ± 0.01 (0.4-0.5) (48) pre flexion 0.4 ± 0.01 (0.3-0.6) (89) 0.5 ± 0.01 (0.4-0.8) (54) flexion - (0) 0.6 (1)

*significantly different (t(0.05)l

Table 10. Morphometries of cultured Sebastes mystinus and S. chrysomelas larvae (mean± 1 S.E. and range). 76

Proportion s. mystinus (n) s. chrysomelas (n) p

*Snout-Anus 30.8 .± 0.18 60 35.1 .± 0.18 48 <0.000 Length (28. 0 - 34.0) (28.0 - 34. 0)

*Head 18.1 .± 0.13 60 17.5 .± 0.17 47 0. 003 Length (16. 0 - 20. 0) (15.0 - 21. 0)

*Body Depth at 14.0 .± 0.16 59 l3 .4 .± 0.12 47 0.010 Pectoral Fin (11. 0 -17.0) (12. 0 -15. 0)

Body Depth at 11.4 + 0.20 55 11.5 .± 0.19 46 0.530 Anus (09.0 - 14. 0) (09. 0 - 15. 0)

*Pectoral Fin 11.2 .± 0.19 55 10.1 + 0.15 45 <0.000 Length (09.0 - 15. 0) (08.0 - 14. 0)

*significantly different (t(0.01))

Table 11. Morphological proportions of 1-2 day old cultured S. mystinus and S. chrysomelas larvae as percentages of notochord length (mean.± 1 S.E. and range). Appendix A: An original Basic program to compare larval pigment patterns within a Sebastes species (Grover, unpub.).

STORE time() to begtime SET color to grt/b RUN dir d:*.dbf ACCEPT •enter dBase filename to compare within no extension' to file SELE A USE &file SET TALK OFF STORE 0 to C1,C2,C3,C4,CS,C6,C7,C8,C9,C10 STORE 0 to C11, C12,C13,C14,C15,C16,C17,C18,C19,C20 STORE 0 to C21,C22,C23,C24,C25,C26,C27,C28,C29,C30,C31 STORE 1 to var STORE 31 to mtotchar STORE 0 to sumpairs &&sum joint /31 STORE 0 to sumsquares &&sum squared joint/31 STORE 0 to tcompare &&total number of comparisons DO WHILE var <=31 STORE 'B'+ltrim(str(var,2)) to varB STORE 'C'+ltrim(str(var,2)) to varC STORE &varB to &varC STORE 1 + var to var END DO STORE 1 to var, recpos, recnoc STORE 0 to mjoint SKIP STORE recno() to recnob DO WHILE .not. eof() DO WHILE var <= 31 STORE 'B'+ ltrim(str(var,2)) to varB STORE 'C'+ ltrim(str(var,2)) to varC IF &varB + &varC <> 1 && 1,1 OR 0,0 at that locus STORE 1 + mjoint to mjoint END IF STORE 1 + var to var END DO STORE 1 + tcompare to tcompare STORE mjoint/mtotchar + sumpairs to sumpairs STORE (mjoint/mtotchar)*(mjoint/mtotchar) + sumsquares to sumsquares IF recnoc = recno() - 1 ?recnoc, recno(), 'mjoint chars', mjoint

77 78

END IF STORE 0 to mjoint STORE l to var SKIP STORE recno() to recnoB END DO RECPOS = l + recpos && increment outside search GOTO recpos DO WHILE var <= 31 STORE 'B'+ltrim(str(var,2)) to varB STORE 'C'+ltrim(str(var,2)) to varC STORE & varB to &varC STORE l + var to var END DO STORE l to var SKIP STORE recno() to recnoC END DO STORE time() to endtime SET PRINT ON ? chr ( 07) ? chr ( 07) ? chr ( 07) ? chr ( 07) ? chr ( 07) ? 'Sum joint loci , sumpairs ? 'Sum squared joint loci , sumsquares ? 'Total number of comparisons tcompare Appendix B. An original Basic program to compare larval pigment patterns between two species of Sebastes (Grover, unpub.).

STORE time( ) to begtime SET color to br+/b RUN dir * . dbf ACCEPT •enter first filename to compare without extension• to file1 ACCEPT 'enter second filename to compare without extension• to file2 SELE B USE &file2 SELE A USE &file1 SET TALK OFF STORE 0 to C1,C2,C3,C4,CS,C6,C7,C8,C9,C10 STORE 0 to C11,C12,C13,C14,C15,C16,C17,C18,C19,C20 STORE 0 to C21,C22,C23,C24,C25,C26,C27,C28,C29,C30,C31 STORE 1 to var STORE 0 to sumpairs &&sum joint/31 STORE 0 to sumsquares &&sum squared joint/31 STORE 0 to tcompare &&total number of comparisons DO WHILE var <= 31 STORE 'B'+ltrim(str(var,2)) to varB STORE 'C'+ltrim(str(var,2)) to varC STORE &varB to &varC STORE 1 + var to var END DO STORE 1 to var STORE recno() to recnoA STORE 0 to mjoint STORE 31 to mtotchar DO WHILE .not. eof( ) SELE B GO TOP DO WHILE .not. eof DO WHILE var <= 31 STORE 'B'+ltrim(str(var,2)) to varB STORE 'C'+ltrim(str(var,2)) to varC IF &varB + &varC <> 1 && 1,1 or 0,0 at that locus STORE 1 + mjoint to mjoint END IF STORE 1 + var to var END DO

79 80

STORE recno() to recnoB STORE 1 +tcompare to tcompare STORE mjoint/mtotchar + sumpairs to sumpairs STORE (mjoint/mtotchar)*{mjoint/mtotchar) + sumsquares to sumsquares IF recnoB = 1 ? recnoA, recnoB, 'mjoint chars', mjoint, 'sumsquares', sumsquares END IF SKIP STORE 0 to mjoint STORE 1 to var END DO SELE A SKIP STORE recno() to recnoA DO WHILE var <= 31 STORE 'B'+ltrim(str(var,2)) to varB STORE 'C'+ltrim(str(var,2)) to varC STORE &varB to &varC STORE 1 + var to var END DO STORE 1 to var END DO STORE time( l to endtime SET PRINT ON ? chr ( 07) ? chr(07) ? chr (07) ? chr(07) ? chr ( 07) ? 'sum joint loci , sumpairs ? 'sum squared joint loci , sumsquares ? 'total number of comparisons tcompare Appendix C. References for mystinus-type Sebastes compared in Figures 6-8 and Tables 6 & 7.

Mystinus-type Sebastes Reference

1. Sebastes aleutianus** Matarese et al. 1989 2. Sebastes brevispinis* * Matarese et al. 1989 3. Sebastes diploproa* Matarese et al. 1989 4. Sebastes flavidus* Matarese et al. 1989 5. Sebastes levis* Moser et al. 1977 6. Sebastes melanops** Moreno. 1990 7 . Sebastes mystinus* Wold. In Matarese et al. 1989 8. Sebastes nigrocinctus* Kendall. 1989 9 . Sebastes paucispinis* Moser et al. 1977 10. Sebastes proriger* Kendall and Lenarz. 1987 11. Sebastes ruberrimus* Kendall. 1989 12. Sebastes cons tel latus+ Moser and Butler. 1987 l3. Sebastes entomelas+ Moser and Butler. 1987 14. Sebastes flavidus+ Kendall. 1989 15. Sebastes goodei+ Moser et al. 1977 16. Sebastes melanops+ Kendall. 1989 17. Sebastes miniatus+ Moser et al. 1977 18. Sebastes mystinus+ Wold. In Matarese et al. 1989 19. Sebastes paucispinis+ Moser et al. 1977 20. Sebastes ruberrimus+ Kendall. 1989 21. Sebastes serriceps+ Moser et al. 1977 22. Sebastes umbrosus+ Moser et al. 1977

*full-term 1-2 day old larvae **premature or intraovarian larvae +larvae L4 days of age

8 1 Appendix D. References for chrysomelas-type Sebastes compared in Figures 9-11 and Tables 8 & 9.

Chrysomelas-type Sebastes Reference

1. Sebastes atrovirens* Moreno. 1990 2. Sebastes auriculatus* Stahl-Johnson. 1985 3. Sebastes babcocki* Kendall and Lenarz. 1987 4. Sebastes ca=atus* Moreno. 1990 5. Sebastes caurinus* Stahl-Johnson. 1985 6. Sebastes chrysomelas* Wold. 1991 7 . Sebastes maliger* Matarese et al. 1989 8. Sebastes rastrelliger** Moreno. 1990 9. Sebastes saxicola* Moser et al. 1977 10. Sebastes atrovirens+ Moreno. 1990 11. Sebastes auriculatus+ Stahl-Johnson. 1985 12. Sebastes ca=atus+ Moreno. 1990 13 . Sebastes caurinus1 + Stahl-Johnson. 1985 14. Sebastes caurinus2 + Moser et al. 1977 15. Sebastes chrysomelas+ Wold. 1991 16. Sebastes dallii+ Moser et al. 1977 17 . Sebastes nebulosus+ Kendall. 1989 18. Sebastes rastrellige+ Moreno. 1990 19. Sebastes semicinctus+ Moser et al. 1977

*full-term 1-2 day old larvae **premature larvae +larvae 2.4 days of age

82 Appendix E: PSI comparisons of pigment patterns described for 1-2 day old larvae of cultured Sebastes mystinus and mystinus-type Sebastes lcnown to occur in the area of Monterey Bay (Fig. 6).

Species Cluster Distance Similarity

s. aleutianus** - s. proriger* 1 0.67 0.33 s. ruberrimus* - Cluster 1 2 0.73 0.27 s. mystinus* - s. nigrocinctus* 3 0. 7 5 0.25 s. diploproa * - Cluster 2 4 0. 78 0.22 Cluster 3 - Cluster 4 5 0. 78 0.22 s. flavidus* - s. melanops** 6 0.79 0.21 s. brevispinis** - Cluster 5 7 0.81 0.19 Cluster 6 - Cluster 7 8 0.81 0.19 s. levis* - s. paucispinis* 9 0.83 0.17 Cluster 8 - Cluster 9 10 0.86 0.14

*full-term 1-2 day old larvae **premature or introvarian larvae

83 Appendix F: PSI comparisons of pigment patterns described for larvae of cultured S. mystinus and mystinus­ type Sebastes 2.4 days of age known to occur in the area of Monterey Bay (Fig. 7).

Species Cluste>r Distance Similarity

S. goodei - S. Lcillerrimus 1 0.50 0.50 S. constellatus - S. mystinus 2 0.54 0.46 Cluster 1 - Cluster 2 3 0.60 0.40 S. entomelas - S. umbrosus 4 0.63 0.37 Cluster 3 - Cluster 4 5 0.65 0.35 S. serriceps - S. miniatus 6 0.71 0.29 Cluster 5 - Cluster 6 7 0.76 0.24 S. melanops - Cluster 7 8 0. 78 0.22 S. flavidus - Cluster 8 9 0.82 0.18 S. paucispinis - Cluster 9 10 0.83 0.17

84 Appendix G: PSI comparisons of pigment patterns described for larvae of cultured S. ~stinus and mystinus-type Sebastes 1-2 and L4 days of age known to occur in the area of Monterey Bay (Fig. 8).

SQecies Cluster Distance Similarity

S. goodei - S. ruberrimus 1 0.50 0.50 S. constellatus - S. ~stinus 2 0.54 0.46 Cluster 1 - Cluster 2 3 0.60 0.40 S. entomelas - S. umbrosus 4 0.63 0.37 Cluster 3 - Cluster 4 5 0.65 0.35 S. aleutianus** - S. proriger 6 0.67 0.33 S. nigrocinctus* - Cluster 5 7 0.68 0.32 S. levis* - S. miniatus 8 0.71 0.29 S. ruberrimus* - Cluster 6 9 0.73 0.27 Cluster 7 - Cluster 8 10 0.74 0.26 s. flavidus* - S. flavidus 11 0.75 0.25 S. serriceps - Cluster 9 12 0. 76 0.24 S. melanops - Cluster 10 13 0.77 0.23 S. diploproa* - Cluster 12 14 0.78 0.22 S. mystinus* - Cluster 14 15 0.78 0.22 Cluster 13 - Cluster 15 16 0.79 0.21 S. melanops** - Cluster 11 17 0.79 0.21 S. brevispinis** - Cluster 16 18 0.81 0.19 Cluster 17 - Cluster 18 19 0.82 0.18 S. paucispinis* - Cluster 19 20 0.83 0.17 S. paucispinis - Cluster 20 21 0.85 0.15

*full-term 1-2 day old larvae **premature or introvarian larvae

85 Appendix H: PSI comparisons of pigment patterns described for 1-2 day old larvae of cultured S. chrysomelas and chrysomelas-type Sebastes known to occur in the area of Monterey Bay (Fig. 9).

Species Cluster Distance Similarity s. auriculatus - s. caurinus 1 0.50 0.50 s. babcocki - Cluster 1 2 0.63 0.37 s. rastrelliger - Cluster 2 3 0.65 0. 3 5 s. maliger - Cluster 3 4 0.68 0.32 s. ca=atus - Cluster 4 5 0.69 0.31 s. atrovirens - Cluster 5 6 0. 72 0.28 s. chrysomelas - Cluster 6 7 0.73 0.27 s. saxicola - Cluster 7 8 0.84 0.16

86 Appendix I: PSI comparisons of pigment patterns described for larvae of cultured S. chrysomelas and chrysomelas-type Sebastes L4 days of age known to occur in the area of Monterey Bay (Fig. 10).

Species Cluster Distance Similarity

s. auriculatus - s. caurinus1 1 0.33 0.67 s. nebulosus - Cluster 1 2 0.40 0.60 • 0 s. caur~nuS" - Cluster 2 3 0.50 0.50 s. atrovirens - Cluster 3 4 0.55 0.45 s. rastrelliger - Cluster 4 5 0.57 0.43 s. carnatus - s. chrysomelas 6 0.58 0.42 Cluster 5 - Cluster 6 7 0.60 0.40 s. dallii - Cluster 7 8 0. 70 0.30 s. semicinctus - Cluster 8 9 0.79 0.21

lStahl-Johnson. 1985 2Moser et al. 1977

87 Appendix J: PSI comparisons of pigment patterns described for larvae of cultured S. chrysomelas and chrysomelas-type Sebastes 1-2 and L4 days of age known to occur in the area of Monterey Bay (Fig. 11).

Species Cluster Distance Similarity

s. auriculatus - s. caurinud 1 0.38 0.62 s. auriculatus* - Cluster 1 2 0.44 0.56 s. nebulosus - Cluster 2 3 0.47 0.53 s. caur1.nus". " - Cluster 3 4 0.50 0.50 s. caurinus* - Cluster 4 5 0.55 0.45 s. atrovirens - Cluster 5 6 0.56 0.44 s. chrysomelas - s. ca=atus 7 0.58 0.42 Cluster 6 - Cluster 7 8 0.59 0.41 s. rastrelliger - Cluster 8 9 0.60 0.40 s. babcocki* - s. maliger* 10 0.63 0.37 Cluster 9 - Cluster 10 11 0.65 0. 35 s. ca=atus* - Cluster 11 12 0.67 0.33 s. rastrelliger* - Cluster 1 13 0.68 0.32 s. dallii - Cluster 13 14 0. 71 0.29 s. chrysomelas* - Cluster 14 15 0. 72 0.28 s. atrovirens* - Cluster 15 16 0.72 0.28 s. semicinctus - Cluster 16 17 0. 79 0.21 s. saxicola* - Cluster 17 18 0.83 0.17

lStahl-Johnson. 1985 2Moser et al. 1977 *full-term 1-2 day old larvae

88