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Editorial Manager(tm) for Journal of Fish Biology Manuscript Draft

Manuscript Number: MS 05-338R2

Title: Gape morphology of cod Gadus morhua L., haddock Melanogrammus aeglefinus (L.) and whiting

Merlangius merlangus (L.) through metamorphosis fom larvae to juveniles in the western Irish Sea.

Article Type: Regular paper Section/Category:

Keywords: gadoid; larvae; gape; jaw; development

Corresponding Author: Dr. William Llewelyn Rowlands, BSc, MSc, PhD Corresponding Author's Institution: University of Liverpool

First Author: William Llewelyn Rowlands, BSc, MSc, PhD

Order of Authors: William Llewelyn Rowlands, BSc, MSc, PhD; Mark Dickey-Collas, PhD; Audrey Geffen, PhD; Richard D.M. Nash , PhD

Manuscript Region of Origin:

Abstract: Variations in standard length, gape size and jaw length were studied in larval and juvenile gadoids from 4-70mm. The increase in gape size and jaw length was not linear with respect to standard length. The relationship was best described by segmented regression lines in all three species, with an inflection point at ca. 10.5mm. Gape size and jaw length increased more rapidly in relation to larval length for individuals smaller than this inflection point size. The rate of increase slowed significantly post-inflection, an effect more noticeable in gape size data compared to jaw length data. In each case the inflection point fell in the

intermediate period of development between the larval and juvenile stages, which could be considered as metamorphosis. Published equations that have been used to predict gape size from jaw length lead to the overestimation of gape. New relationships are presented which may be used to predict gape size from measurements of either standard length or upper jaw length in cod, haddock and whiting.

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Dear Dr Craig,

I have re-submitted my manuscript (MS 05-338R1) with the following alterations:

1. Line numbers have been added

2. Authorities have been corrected to Gadus morhua L., Melanogrammus aeglefinus (L.) and Merlangius merlangus (L.)

3. Presentation of statistics in text changed from (GLM, F(2,536) = 8.6, p<<0.001) to (GLM, d.f. = 2, n = 536, P<0.001) in all cases

4. Variables changed as shown below - standard length: LS to LS

upper jaw length: LUJ to LUJ

lower jaw length: LLJ to LLJ

gape size: SG to SG

5. Indents corrected on paragraphs 6. References corrected in the text

7. Tables all converted from excel files to “word tables” so are now all doubled spaced (however they do not look as good)

8. Figures all converted to EPS files

9. Reference list corrected – there was something wrong with the formatting I hope this covers all aspects that needed revising

Yours,

William Rowlands

* Response to referees

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Gape morphology of cod Gadus morhua L., haddock Melanogrammus

1

aeglefinus (L.) and whiting Merlangius merlangus (L.) through

2

metamorphosis from larvae to juveniles in the western Irish Sea.

3 4

W. LL. ROWLANDS*∞, M. DICKEY-COLLAS, A. J. GEFFEN*AND R. D. M. NASH* 5

6

* University of Liverpool, Port Erin Marine Laboratory, Port Erin, Isle of Man, IM9 6JA, British Isles.

7

Netherlands Institute of Fisheries Research, P.O. Box 68, 1970 AB IJmuiden, The Netherlands.

8 9

Variations in standard length, gape size and jaw length were studied in larval and juvenile gadoids from 10

4-70mm. The increase in gape size and jaw length was not linear with respect to standard length. The 11

relationship was best described by segmented regression lines in all three species, with an inflection 12

point at ca. 10.5mm. Gape size and jaw length increased more rapidly in relation to larval length for 13

individuals smaller than this inflection point size. The rate of increase slowed significantly post- 14

inflection, an effect more noticeable in gape size data compared to jaw length data. In each case the 15

inflection point fell in the intermediate period of development between the larval and juvenile stages, 16

which could be considered as metamorphosis. Published equations that have been used to predict gape 17

size from jaw length lead to the overestimation of gape. New relationships are presented which may be 18

used to predict gape size from measurements of either standard length or upper jaw length in cod, 19

haddock and whiting.

20 21 22

Key words: gadoid; larvae; gape; jaw; development.

23

Present address: Department of Biology, University of Bergen, P.O. Box 7800, 5020 Bergen, Norway.

Present address: Institute of Marine Research, P.O. Box 1870, N-5817 Bergen, Norway.

Author to whom correspondence should be addressed. Tel: +44 (0) 1624 831037; Fax: +44 (0) 1624 831001; email: [email protected]

Manuscript

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24 25

INTRODUCTION 26

27

Cod Gadus morhua L., haddock Melanogrammus aeglefinus (L.) and whiting 28

Merlangius merlangus (L.) are commercially important white-fish which spawn 29

between March and May in the Irish Sea (Bal, 1941; Wheeler, 1969; Nichols et al., 30

1993; Brander, 1994). Due to similarities in spawning times and locations there is a 31

considerable overlap in the distribution of the pelagic larval stages of these three 32

species (Fox et al., 1997). This distributional overlap along with the utilization of 33

similar planktonic prey items through development (Last, 1978; Robb & Hislop, 34

1980; Economou, 1991) implies a high degree of inter- and intra-specific competition 35

for food; as previously suggested by Kane (1984) with >75% overlap between larval 36

gadoid diets.

37 38

The extent of competition for food is directly related to food availability (Last, 39

1978), and the size distribution of available prey (Bremigan & Stein, 1994). The array 40

of these available prey items is defined by prey detection, swimming ability, strike 41

ability and mouth size of the larvae (Sabatés & Saiz, 2000), all of which develop with 42

age (Hunter, 1980).

43 44

This study compared variations in gape size between species, as this is a major 45

factor limiting the maximum size of prey consumed by larvae (Arts & Evans, 1987;

46

Bremigan & Stein, 1994). Larval development proceeds through different growth 47

phases, leading to changes in allometric growth patterns (Osse & van den Boogaart, 48

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1995; van Snik et al., 1997). If gape size increases in this manner with body length 49

then the rate of gape increase could be partially responsible for specific differences in 50

food size selection and thus niche prey size (Hunter, 1980) between the larvae of 51

different species. This is so as at the start of exogenous feeding larvae with larger 52

gape size have been shown to have faster growth rates (Shirota, 1970, 1978). The 53

faster growth rate is due to the larvae’s ability to take a larger size of prey with leads 54

to increased growth efficiency (Kerr, 1971a, b). A larger gape size also leads to a 55

larger size selection of prey available to the larvae and this prey niche size has been 56

shown to increase with larval development, though is not constant between 57

developmental periods or between different species (Pepin & Penny, 1997). The 58

importance the increase in prey niche size is that intraspecific competition between 59

early and late spawned members of the same year class will be reduced due to the 60

increase prey available (Robb & Hislop, 1980; Pepin & Penny, 2000). Therefore, with 61

the ability to both ingest a larger size of prey and have a larger prey niche size, the 62

effects of intra- and interspecific competition for food between larvae would be 63

reduced, increasing chances of survival. However, Pepin & Penny (1997) observed 64

that the rate of increase in prey size was slower than the increase in the physical 65

capacity to ingest larger prey. This suggests that perception and vulnerability of prey 66

along with environmental factors are further elements required for comparative 67

studies of larval feeding ecology.

68 69

To address the physical and morphological limitations, this study examines 70

and compares the development of gape size between cod, haddock and whiting during 71

larval development. Although work has previously been carried out on the gape sizes 72

of marine larvae (Shirota, 1970; Schael et al., 1991; Munk, 1997; Sabatés & Saiz, 73

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2000), little has been done on comparisons in relation to metamorphosis in gadoid 74

larvae to juveniles. This is an important life history phase, as during this period larvae 75

undergo rapid internal and external changes (Kjørsvik et al., 1991; Osse & van den 76

Boogaart, 1995; Fuiman & Higgs, 1997). Morphological comparisons between the 77

species were undertaken through the gape measurement of these gadoids and thus 78

probable overlaps in available prey items evaluated.

79 80

MATERIALS AND METHODS 81

82

SAMPLE COLLECTION AND PRESERVATION 83

84

Samples of fish larvae were collected on two occasions (April and May 2001) 85

in the northwestern Irish Sea from the Department of Agriculture and Rural 86

Development – Northern Ireland (DARD) research vessel, the M.R.V. Lough Foyle.

87

The full range of developmental stages, yolk-sac larvae to juveniles, were sampled 88

with a Hydrobios Multiplankton (MP) Sampler with 280µm mesh (Weikert & John, 89

1981) to capture yolk-sac larvae in both April and May, a MOCNESS sampler 90

(Multiple Opening and Closing Net and Environmental Sensing System (Wiebe et al., 91

1976; Roe & Shale, 1979)) with 1000µm mesh to sample larger larvae in May only 92

and a German pelagic juvenile trawl in May only to sample juveniles. The first two 93

samplers were towed for 20 minutes using a stepwise oblique profile at 3 knots (1.5 94

m.s-1). The later gear was towed for 20 minutes at between four and five knots (2-2.6 95

m.s-1). Over a twenty-four hour period the MP was deployed with two replicates every 96

four hours, the MOCNESS six times, and the juvenile pelagic trawl only once. This 97

sampling strategy was part of a larger project and these deployments were carried out 98

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at four different stations in the northwestern Irish Sea (Fig. 1). The high numbers of 99

deployments gave a relatively large catch of larval and juvenile gadoids.

100 101

All samples were preserved immediately in 4% buffered (sodium acetate 102

trihydrate) formaldehyde to reduce larval shrinkage (Blaxter, 1971; Hay, 1981).

103 104

MORPHOLOGICAL AND DEVELOPMENTAL MEASUREMENTS 105

106

Standard lengths of cod, haddock and whiting larvae were measured in the 107

laboratory, either under a dissecting microscope with graticule (with a precision of 108

0.02mm), or in the case of larger juveniles with vernier calipers (with a precision of 109

0.05mm).

110 111

Gape sizes were calculated using a method similar to that of Wong & Ward 112

(1972), which allowed the limiting dimension to be measured, in this case with 113

differently sized disposable plastic pipette tips ranging from 0.3mm to 10mm in 114

diameter. The pipette tips were placed under a microscope and scored along their 115

length using a scalpel at 0.5mm intervals. The scored marks were then rubbed with 116

permanent ink to leave a clearly identifiable line. The diameter of the pipette tip at 117

each scored interval was measured using a dissecting microscope with graticule to 118

give a range of sizes (diameters) along the pipette tip with an accuracy of 0.05mm.

119 120

Gape size was measured by holding a larva under the microscope, while 121

inserting a calibrated pipette tip into the mouth until a jaw angle of 90º was achieved.

122

The diameter of the cone at the scored mark where the front tips of the upper and 123

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lower jaws touched the cone was taken to be the gape. The measurement was made at 124

a mouth angle of 90º as this has been assumed to give the maximum gape for larval 125

fish (Shirota, 1970). Care had to be taken not to insert the cones too far, as this caused 126

the pipette tip to reach the back of the mouth before the jaws were fully opened thus 127

giving false readings (Arts & Evans, 1987). In order to avoid this problem, a slightly 128

larger sized tip was used if a tip went too far into the mouth.

129 130

The upper and lower jaw lengths of the larvae were measured under a 131

dissection microscope with graticule and defined as the distance from the tip of the 132

jaw to where the upper and lower jaws joined. Jaw lengths were measured with the 133

mouth in the closed position.

134 135

The measurement of the jaws also allowed gape sizes to be calculated using 136

equations given by Shirota (1970) and later modified by Guma’a (1978);

137 138

( )

UJ

G

L

S = 2 ∗

(Equation 1) (Shirota, 1970)

139

(

UJ2 LJ2

)

G

L L

S = +

(Equation 2) (Guma’a, 1978)

140 141

Where the calculated gape size (SG), was obtained from the length of the upper jaw 142

(LUJ) and the length of the lower jaw (LLJ).

143 144

Basic developmental features of each larva were also recorded using external 145

and internal morphology (Balon, 1975; Timmermans, 1987; Kjørsvik et al., 1991;

146

Osse & van den Boogaart, 1995). A score was allocated to each developmental 147

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observation (Table I) which, when totaled, gave a generalized index of larval 148

development for each individual.

149 150

STATISTICAL ANALYSIS 151

152

Due to the allometric relationships observed between larval length and 153

measured structures, data were logarithmically transformed to give linear 154

relationships allowing ease of comparison between species (Peters, 1983). However, 155

the data best fitted a segmented regression, based on examination of the residuals 156

(Peck et al., 2005). The linear regression showed a clear trend in the plotted residuals 157

(Fig. 2 (Insert A)) suggesting a systematic error in the relationship, while the plotted 158

residuals of the segmented relationship (Fig. 2 (Insert B)) showed no pattern, 159

suggesting a good fit to the data. This was shown to be the case for all morphological 160

measurements on all three species, and so a segmented regression was used for all 161

subsequent analyses.

162 163

The points of inflection for the segmented relationships were found using the 164

computer program PRISM® (produced by GRAPHPAD® (www.graphpad.com)), 165

which calculated the residual sum of squares (RSS), testing each inflection point 166

iteratively to obtain the lowest RSS value (Somerton, 1980).

167 168

The significance of each segmented regression line was tested and 169

comparisons between regression lines were made using the General Linear Model 170

(GLM) assuming normality and homogeneity of variance. All calculations were made 171

using MINITAB® release 13.1 (www.minitab.com).

172

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173

RESULTS 174

175

GAPE SIZE/STANDARD LENGTH RELATIONSHIP 176

177

Gape size increased allometrically with standard length and when transformed 178

using the natural log, a segmented relationship with a single inflection point was 179

observed (Fig. 3). Although a single line linear regression could be fitted to these data, 180

it obscured the true relationship between gape size and fish length (Table IIi – shown 181

by the lower F statistic values) . The inflection points were located at a standard 182

length of 9.84mm for whiting, 10.21mm for haddock and 11.72mm for cod. The 183

differences between species were not significant, suggesting a global gadoid 184

breakpoint of 10.56mm.

185 186

In all three species the slope of the regression of gape size on length for fish 187

pre-inflection point was >1 (positive allometry) and post-inflection point the slope 188

was <1 (negative allometry), (Table IIi). In all cases the relationship pre- and post- 189

inflection point was significantly different (GLM, d.f. = 2, n = 141, P<0.001; d.f. = 2, 190

n = 258, P<0.001; d.f. = 2, n = 553, P<0.001, for cod, haddock and whiting 191

respectively).

192 193

Among smaller larvae (pre-inflection point) the gape size of whiting increased 194

at a significantly slower rate than that of haddock (GLM, d.f. = 2, n = 520, P<0.001), 195

while the rate of gape size increase of cod fell between that of whiting and haddock 196

and was not significantly different to either. In larger larvae (post-inflection point) the 197

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gape size of whiting increased at a significantly faster rate with length than for both 198

haddock and cod (GLM, d.f. = 2, n = 433, P<0.001); again no difference was seen 199

between cod and haddock.

200 201

JAW LENGTH/STANDARD LENGTH RELATIONSHIP 202

203

Upper jaw length and standard length data (Fig. 4) also showed an allometric 204

relationship and on transformation (natural log), a segmented regression explained 205

more variance compared to a standard single linear regression (Table IIii shown by 206

the higher F statistic values). A significant reduction in the rate of jaw length increase 207

with standard length post-inflection point was observed for all species (GLM, d.f. = 2, 208

n = 141, P<0.001; d.f. = 2, n = 259, P<0.001; d.f. = 2, n = 552, P<0.001, for cod, 209

haddock and whiting respectively). Inflection points for cod, haddock and whiting 210

occurred at standard lengths of 10.51mm, 8.95mm and 10.36mm respectively, with no 211

significant difference between species suggesting a global gadoid inflection point at 212

9.94mm.

213 214

Pre-inflection point regression comparisons between species, showed the rate 215

of jaw length increase with standard length of haddock significantly greater (GLM, 216

d.f. = 2, n = 602, P<0.001) than for either cod or whiting. Post-inflection point 217

regression comparisons showed haddock to have the slowest rate of jaw length 218

increase (GLM, d.f. = 2, n = 536, P<0.001).

219 220

GAPE SIZE/JAW LENGTH RELATIONSHIP 221

222

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The upper jaw length was used for analysis, as it was the easier dimension to 223

measure, and therefore most reliable. When compared, upper jaw length and gape size 224

again produced a curved relationship, which was transformed using natural logs. A 225

segmented relationship with single inflection point (Fig. 5) was a better fit to the data 226

than a single linear regression (F-statistic greater in segmented relationships - Table 227

IIiii). The positions of these inflection points were compared and no differences were 228

observed between species, occurring at a jaw length of 1.27mm.

229 230

For each of the three species the rate of gape size increase with jaw length 231

decreased after the inflection point (GLM, d.f. = 2, n = 188, P<0.001; d.f. = 2, n = 232

317, P<0.001; d.f. = 2, n = 632, P<0.001, for cod, haddock and whiting respectively), 233

changing from positive to negative allometry. Between species, the slope of the 234

regression lines below the inflection point showed no differences. However, 235

regressions above the inflection point suggested that gape size increased at a 236

significantly faster rate with jaw length for cod compared to either haddock or whiting 237

(GLM, d.f. = 2, n = 411, P<0.05), with no difference between the haddock and 238

whiting.

239 240

MODELS FOR ESTIMATING GAPE 241

242

Gape size was calculated from jaw length, following equations in Shirota 243

(1970) and Guma’a (1978) and compared to the actual measurements of gape size.

244

The calculated gape sizes were significantly larger than actual gape measurements 245

(GLM, d.f. = 2, n = 1801, P<0.001), with post-inflection point data (those >9.84mm – 246

taken from Fig. 4) showing increased over-estimation.

247

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248

From data in this study, individual models converting standard length (LS), 249

upper jaw length (LUJ) and lower jaw length (LLJ) to gape size (SG) could be calculated 250

and are shown below. The associated variables (a and b) for each gadoid species pre- 251

and post-inflection point are given in Table III:

252 253

b L

S

G

= (

Sa

) *

(Equation 3) 254

b L

S

G

= (

UJa

) *

(Equation 4) 255

b L

S

G

= (

LJa

) *

(Equation 5) 256

257

Each model is subject to size constraints due to the segmented nature of the 258

data and the size range of larvae used. Pre-inflection equations for all models are 259

limited to larvae of standard lengths and upper and lower jaw lengths below the size at 260

inflection. The standard length model, post-inflection equations have an upper size 261

limit of 50mm for cod, and 65mm for haddock and whiting. The upper and lower jaw 262

length models, post-inflection equations have an upper limit of 4.5mm for cod, 6.5mm 263

for haddock and 7.5mm for whiting.

264 265

LARVAL DEVELOPMENT 266

267

The relationship between developmental index and larval length (natural log 268

standard length – Fig. 6) was well fitted by a segmented regression model with two 269

inflection points (ANOVA, d.f. = 2, n = 185, P<0.001; d.f. = 2, n = 255, P<0.001; d.f.

270

= 2, n = 482, P<0.001, for cod, haddock and whiting respectively). For cod, haddock 271

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and whiting, development fell into three distinct categories named as larval, 272

intermediate and juvenile stages. The developmental index increased rapidly with 273

length following the first inflection point, suggesting that the transition from the larval 274

to intermediate stages was linked to the start of metamorphosis (Fig. 6). This occurred 275

at a standard length of 8.21mm for cod, 8.13mm for haddock and 7.71mm for whiting.

276

These were not found to be significantly different suggesting a general global gadoid 277

size of ca. 8mm could be considered as the start of metamorphosis.

278 279

The developmental index was calculated primarily from changes in fin and 280

stomach characteristics, and their period of most rapid development (intermediate 281

stage) ended at a standard length of 15.61mm for cod, 12.85mm for haddock and 282

13.67mm for whiting. Once again no significant difference was found between these 283

values (partly due to the large size variation in the data) implying a general global 284

gadoid size of ca. 14mm.

285 286

DISCUSSION 287

288

The relationships between gape size and standard length, and jaw length and 289

standard length for cod, haddock and whiting changed from positive to negative 290

allometry during development. A change in allometry has previously been shown to 291

occur in the head and tail of larvae, due to initial high-energy investment for primitive 292

functions and essential organs (Osse & van den Boogaart, 1995). In this study only 293

mouth morphology was studied, therefore what is the advantage of reducing the rate 294

of gape and jaw length increase? An explanation could be that newly hatched larvae 295

require rapid growth. The need for rapid growth is to avoid predation, as the smaller 296

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the larvae the more likely they are to be predated (Scharf et al., 2000; Aljetlawi et al., 297

2004) and to allow them to obtain a greater size spectrum of prey rapidly, to avoid 298

competition from more recently hatched larvae (Sabatés & Saiz, 2000). The 299

implication to gape size is that it has been shown that a larger gape size leads to larger 300

prey items (Robb & Hislop, 1980), which in turn leads to more rapid growth (Hunter, 301

1980). Therefore the greater the ratio of gape size to body length the greater the 302

ability of a larva to take large prey, and grow more rapidly. A high gape size to body 303

length ratio would therefore be especially beneficial for first feeding larvae and 304

explain the observed positive allometry pre-inflection point in gape size and standard 305

length data. However, once at the juvenile stage the need for rapid growth is lessened 306

due to a reduction in competition for food, due to a shift to larger prey items (Pearre, 307

1986; Sabatés & Saiz, 2000), and reduced predation pressures (Scharf et al., 2000).

308

Thus the energetic expenditure for rapid gape increase could be reduced and 309

apportioned to other aspects of development.

310 311

A segmented relationship leading to a reduction in the rate of mouth growth 312

(as jaw length) with increasing body length was reported previously in a number of 313

marine larvae by Shirota (1978), where the point of inflection was seen to differ 314

between species. However, in this study the inflection point occurred at the same 315

standard length for all species for gape size, and upper jaw length data, implying a 316

general gadoid model, certainly for these gadoids in the Irish Sea. In these fish the 317

inflection points occurred at a standard length corresponding to the metamorphic 318

period of development (between ca. 8mm and ca. 14mm). This may also be seen in 319

other gadoids as Osse & van den Boogaart (1995) suggested that there was similarity 320

in particular allometries at an equal size range of fish larvae of distantly related taxa 321

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and Mandali de Figueiredo (2003) showed that the larvae of two gadoid species 322

(whiting and rockling), had identical gape size to body length ratios. These 323

observations suggest that a global gadoid model could be assumed, where gape and 324

jaw allometry could be used as aids in the developmental staging of larvae into this 325

metamorphic stage, a poorly defined area in the life history of fish (Copp & Kováč, 326

1996).

327 328

Although similarities in standard lengths at allometric inflection points, and 329

developmental stages suggest a global gadoid model, some differences between 330

species did occur. The rate of gape increase with standard length both pre- and post- 331

inflection point were shown to differ between species as suggested previously by 332

Robb & Hislop (1980). Whiting were shown to have a slower rate of gape size 333

increase pre-inflection compared to haddock but a greater rate post-inflection 334

compared to both cod and haddock. Due to the slowest rate of gape increase, whiting 335

had the smallest gape size at the inflection point, while those of cod and haddock were 336

larger and did not differ (cod: 1.57mm; haddock: 1.43mm; whiting: 1.25mm – 337

obtained from Fig. 3 and equations in Table III). However, post inflection the gape 338

size of whiting rapidly became larger than that of both cod and haddock, suggesting 339

that in terms of gape size whiting differed consistently from both cod and haddock.

340

The similarities in gape size of cod and haddock throughout development could imply 341

that inter-specific competition would be high due to a similar prey niche size.

342

However, cod larvae take larger sized prey than both haddock and whiting, (Robb &

343

Hislop, 1980), and haddock in turn have been shown to be the least selective of these 344

larvae, having a tendency to go for more slow moving prey items (Economou, 1991).

345

This suggests that although gape size can give an indication of maximum prey size it 346

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cannot be used by itself to determine prey niche size as other factors such as predator 347

and prey behavior will also influence prey selection (Pepin & Penny, 1997; Scharf et 348

al., 2000). A study of prey niche size requires gut analysis along with gape size 349

analysis, and knowledge of the prey available in the environment, especially for low 350

concentrations of larger prey which may lead to underestimating niche sizes (Munk, 351

1997; Gonzalez-Quiros & Anadon, 2001).

352 353

Gape size is an important parameter to measure for studies of feeding ecology 354

and potentially for studies of larval development. Gape size is often difficult to 355

measure directly and is often calculated from jaw measurements. This study compared 356

measured gape size to gape size calculated from the equations of Shirota (1970) and 357

Guma’a (1978), and showed clearly that gape sizes derived from the equations 358

consistently overestimated the actual maximum gape (as measured directly), being 359

especially pronounced after the point of inflection. This observation is directly 360

comparable with the work of Shirota (1970) who estimated gape size up to a larval 361

length of 30mm, well beyond the inflection points found in this study. As both mean 362

and maximum prey sizes are a proportion of gape size (Robb & Hislop, 1980), this 363

discrepancy would suggest that larvae could take a greater size distribution of prey 364

items than was actually the case. If used in comparisons such as with prey size (as in 365

Munk (1997)) an overestimation of prey available will occur and affect the prediction 366

of larval survival in relation to prey present.

367 368

Shirotas’ (1970) and Guma’as’ (1978) calculations rely on a single 369

relationship to derive gape size from jaw length. When post-inflection point data was 370

compared to pre-inflection point data from this study there was a 25% ± 5% reduction 371

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in the rate of jaw length increase compared to a 50% ± 5% reduction in the rate of 372

gape size increase for the same increase in standard body length. Thus the real 373

segmented allometric effect was not fully translated into the gape size calculations of 374

Shirota (1970) and Guma’a (1978), leading to overestimations of gape size, especially 375

during both the later intermediate and juvenile stages. Two non-linear models (one 376

pre- and one post- inflection) for each gadoid species that incorporate the relative 377

changes of gape size in relation to either jaw length or fish length produces better 378

estimates where gape size cannot be measured directly. This method of gape size 379

calculation on unpreserved larvae would have to be carried out with caution as 10%

380

shrinkage has been observed in the standard length of other gadoid species preserved 381

in formalin (Porter et al., 2001).

382 383

In conclusion, gape size can be, a useful tool in establishing prey niche size of 384

fish larvae, and in helping to developmentally stage larval fish. However, if gape size 385

is calculated from jaw length, the allometric relationships as larvae grow and develop 386

must be incorporated. Although gape sizes and rates of gape size increase have been 387

shown to vary between species, similarities suggesting the existence of a global 388

gadoid model were also observed. These similarities were related to larval 389

development, where in all species, larval length at changes in jaw and gape allometry 390

did not differ significantly. In every case these changes in allometry occurred in the 391

same period of development, the intermediate stage, equating to the period of most 392

rapid development implying metamorphosis. This was found not to differ between 393

species implying a general gadoid length for the start of metamorphosis at 8mm, with 394

rapid development ceasing at a length of 14mm. Further experimentation of interest 395

would therefore be the larval study of other gadoid species to see if they too adhered 396

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to these developmental patterns. Another interesting comparison would be the 397

application of the newly derived gape calculation equations to adult cod, haddock and 398

whiting.

399

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TABLES

TABLE I. Developmental features and assigned scores for the determination of a developmental index for larval fish.

Developmental features based on observations from Balon (1975), Timmermans (1987), Kjørsvik et al. (1991) and Osse & van den Boogaart (1995), external features represented above the dashed line and internal features below.

TABLE II. Regression analysis of cod, haddock and whiting for i) natural log transformed gape size (SG) versus standard length (LS) data; ii) natural log transformed upper jaw length (LUJ) versus standard length (LS) data; iii) natural log transformed gape size (SG) versus upper jaw length (LUJ) data when carried out as a single and segmented regression.

Score Developmental Feature 1 Yolk-sac larvae with mouth closed 2 Yolk-sac larvae with mouth open 3 Exogenous feeding larvae – finfold complete 4 Exogenous feeding larvae – finrays visible 5 Exogenous feeding larvae – all fins present 6 Teeth not visible in mouth

7 Teeth visible in mouth 8 Incipient gut 9 Gut looped but stomach not visible

10 Stomach visible 11 Pyloric caeca visible Table

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Species Regression type r2 F d.f. Regression equation

i) Cod Single 0.94 2377 140 ln(SG) = 1.019ln(LS)-2.281

Segmented 0.98 5350 138 ln(SG) = 1.376ln(LS)-2.938 Pre-inflection ln(SG) = 0.678ln(LS)-1.220 Post-inflection Haddock Single 0.94 3752 257 ln(SG) = 0.923ln(LS)-2.054

Segmented 0.98 13486 255 ln(SG) = 1.500ln(LS)-3.128 Pre-inflection ln(SG) = 0.654ln(LS)-1.164 Post-inflection Whiting Single 0.97 15610 552 ln(SG) = 0.924ln(LS)-2.005

Segmented 0.98 30285 550 ln(SG) = 1.331ln(LS)-2.824 Pre-inflection ln(SG) = 0.803ln(LS)-1.616 Post-inflection ii) Cod Single 0.96 3708 140 ln(SG) = 0.961ln(LUJ)+0.003

Segmented 0.97 5265 138 ln(SG) = 1.134ln(LUJ)+0.085 Pre-inflection ln(SG) = 0.773ln(LUJ)+0.160 Post-inflection Haddock Single 0.98 11080 261 ln(SG) = 0.911ln(LUJ)+0.032

Segmented 0.99 19512 259 ln(SG) = 1.100ln(LUJ)+0.043 Pre-inflection ln(SG) = 0.758ln(LUJ)+0.136 Post-inflection Whiting Single 0.98 21710 554 ln(SG) = 0.893ln(LUJ)+0.011

Segmented 0.98 27711 552 ln(SG) = 1.063ln(LUJ)+0.039 Pre-inflection ln(SG) = 0.807ln(LUJ)+0.103 Post-inflection iii) Cod Single 0.97 6705 187 ln(LUJ) = 1.070ln(LS)-2.410

Segmented 0.97 9163 185 ln(LUJ) = 1.198ln(LS)-2.628 Pre-inflection ln(LUJ) = 0.949ln(LS)-2.042 Post-inflection Haddock Single 0.97 11920 316 ln(LUJ) = 1.038ln(LS)-2.302

Segmented 0.99 31284 314 ln(LUJ) = 1.325ln(LS)-2.810 Pre-inflection ln(LUJ) = 0.872ln(LS)-1.751 Post-inflection Whiting Single 0.98 37740 628 ln(LUJ) = 1.040ln(LS)-2.277

Segmented 0.98 62172 626 ln(LUJ) = 1.221ln(LS)-2.627 Pre-inflection ln(LUJ) = 0.991ln(LS)-2.123 Post-inflection

The regression equation, coefficient of determination (r2), and associated ANOVA with F statistic (F) and degrees of freedom (d.f.) are given. All significance was at the p<<0.001 level.

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TABLE III. Associated variables used for the calculation of larval gape size, when using i) standard length, ii) upper jaw length and iii) lower jaw length measurements.

variable Species Size range (mm)

a b

i) Cod <10.56 1.376 5.297 x 10-2 >10.56 0.768 2.952 x 10-1 Haddock <10.56 1.500 4.381 x 10-2 >10.56 0.654 3.122 x 10-1 Whiting <10.56 1.331 5.937 x 10-2 >10.56 0.803 1.987 x 10-1 ii) Cod <1.27 1.134 1.089

>1.27 0.773 1.174

Haddock <1.27 1.100 1.044

>1.27 0.758 1.146

Whiting <1.27 1.063 1.040

>1.27 0.807 1.109

iii) Cod <1.27 1.030 1.207

>1.27 0.672 1.439

Haddock <1.27 1.050 1.095

>1.27 0.782 1.246

Whiting <1.27 1.010 1.078

>1.27 0.804 1.213

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1

FIGURE CAPTIONS 1

2

FIG. 1. Sample area in the western Irish Sea, for the collection of larval and juvenile cod, 3

haddock and whiting during April and May of 2001.

4 5

FIG. 2. Natural log plot of gape size and standard length for cod. A plot of the residuals 6

from a linear regression are shown in Insert A, and those for a segmented regression shown 7

in Insert B 8

9

FIG. 3. Natural log transformed standard length and gape size for a) cod, b) haddock and c) 10

whiting. The inflection point position at standard length (solid vertical line) ± 95% CI’s 11

(dashed vertical line) for cod (2.46 ± 0.16), haddock (2.32 ± 0.06) and whiting (2.29 ± 12

0.05) is shown.

13 14

FIG. 4. Natural log transformed standard length and upper jaw length for a) cod, b) 15

haddock and c) whiting. The inflection point position at standard length (solid vertical line) 16

± 95% CI’s (dashed vertical line) for cod (2.34 ± 0.29), haddock (2.34 ± 0.11) and whiting 17

(2.19 ± 0.10) is shown.

18 19

FIG. 5. Natural log transformed upper jaw length and gape size for a) cod, b) haddock and 20

c) whiting. The inflection point position at upper jaw length (solid vertical line) ± 95%

21

CI’s (dashed vertical line) for cod (0.21 ± 0.21), haddock (0.27 ± 0.12) and whiting (0.25 ± 22

0.12) is shown.

23 24 Figure Captions

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2

FIG. 6. Relationship between developmental index and natural log transformed standard 25

length data for a) cod, b) haddock and c) whiting. Inflection points at standard length for 26

the change of larval stage are shown as solid vertical lines, ± 95% CI’s shown as dashed 27

vertical lines. Shaded area equates to the region of “intermediate” stage (circle symbols) 28

after the larval stage (cross symbols) and before the juvenile stage (triangular symbols).

29 30 31

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Figure

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Figure

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Figure

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Figure

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Figure

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Figure

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