Impact of a population bottleneck on symmetry and

genetic diversity in the northern elephant seal

A. RUS HOELZEL,* R. C. FLEISCHER,*† C. CAMPAGNA,‡ B. J. LE BOEUF§ & G. ALVORD–

*Department of Biological Sciences, Durham University, South Road, Durham DH1 3LE, UK
†Department of Zoological Research, National Zoological Park, Smithsonian Institution, Washington, DC 20 008, USA
‡Centro Nacional Patagonico, CONICET, 9120 Puerto Madryn, Chubut, Argentina
§Department of Biology, University of California, Santa Cruz, CA 95 064, USA
–Data Management Services Inc., National Cancer Institute, FCRDC, Frederick, MD 21 702, USA

Keywords: Abstract
bottleneck; The northern elephant seal (NES) suffered a severe population bottleneck
fluctuating asymmetry; towards the end of the nineteenth century. Theoretical expectations for the
genetic diversity; impact of population bottlenecks include the loss of genetic diversity and a loss
marine mammal. of fitness (e.g. through a disruption of developmental stability); however,
there are few direct demonstrations in natural populations. Here, we report on
the comparison of archive samples collected prior to and following the NES
population bottleneck. Measures of genetic diversity show a loss of variation
consistent with expectations and suggest a strong disruption in the pattern of
allele frequencies following the bottleneck. Measures of bilateral characters
show an increase in fluctuating asymmetry.

Introduction after a bottleneck event in one of a number of extant

The impact of population bottlenecks on the diversity
and fitness of natural populations is an important
question in the context of both evolutionary process
and conservation biology. The theoretical expectation
with respect to genetic diversity can be calculated based
on the stochastic loss of alleles, and the probability of
recombining alleles that are identical by decent (e.g. Nei
et al., 1975). Experimental studies have tended to sup-
port theoretical expectations, although they also illus-
trate the large variance in outcome among different
bottleneck events, and show that the impact on allelic
diversity is more predictable than the impact on hetero-
zygosity (e.g. Leberg, 1992). However, most studies
assessing the impact of known and putative bottleneck
events in natural populations are based on comparative
analyses, either between species or between conspecific
populations. Direct comparisons of the same population
before and after a bottleneck are relatively rare. Among
these, essentially all have compared variation before and
Correspondence: A. Rus Hoelzel, Department of Biological Sciences,
Durham University, South Road, Durham, DH1 3LE, UK.
Tel.: +44-(0)191 3747745; fax: +44-(0)191 3742417;
e-mail: a.r.hoelzel@durham.ac.uk
populations of a species (Britten & Brussard, 1996;
Bouzat et al., 1998; Groombridge et al. 2000; Whitehouse
& Harley 2001). However, it is often difficult to control
for the possible influence of immigration, which can
have a substantial effect on the rate of recovery of
genetic variation in post-bottleneck populations (e.g.
Ratnaswamy et al., 1999; Keller et al. 2001).
In this study, we compare pre- and post-bottleneck
populations in a species [the northern elephant seal
(NES), Mirounga angustirostris] where only one popula-
tion was left after the bottleneck, so that any post-
bottleneck diversity must have survived the bottleneck
event. In addition to the direct genetic comparison, we
also analyse skulls from the prebottleneck sample for
measures of bilateral symmetry, and compare them with
measurements from post-bottleneck skulls and from a
congeneric species, the southern elephant seal (SES)
(Mirounga leonina). In this way, we test the prediction
that a severe bottleneck can impact on developmental
stability, as indicated by a disruption of fluctuating
asymmetry (FA; Waddington, 1957; Leamy, 1984;
Bryant et al., 1986; Hutchison & Cheverud, 1995). It has
long been considered that FA could be used as an
indicator of fitness related to developmental stability
(Beardmore, 1960; Soule´ , 1967; Parsons, 1992).
Among

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568 A. RUS HOELZEL ET AL.

the three types of asymmetry (directional, fluctuating

and antisymmetry) only FA has been suggested to result
from poorly coadapted gene complexes, and therefore to
be useful as a measure of developmental stability. This
builds on an earlier analysis of NES skulls (Hoelzel, 1999)
that showed greater variability of morphometric charac-
ters in post than pre bottleneck skulls, consistent with
earlier experiments (primarily on house flies) and
predictions related to the disruption of non–additive
genetic interactions following bottleneck events (see
Bryant et al., 1986).
The NES was heavily exploited during the nineteenth
century and reduced to a bottleneck population size
estimated to be only 10–30 individuals (Bartholomew &
Hubbs, 1960; Hoelzel et al., 1993). The hunt began
around 1810 and became commercially unviable by the
1860s as a result of overexploitation. However, NES
continued to be taken intermittently, including over 400
between 1880 and 1884 (Townsend, 1885). The popu-
lation has now recovered to more than 100 000 seals
(Stewart et al., 1994). This provides the basis for a direct
assessment of the impact of a severe bottleneck on
genetic diversity and a measure of developmental
stabil- ity in a natural population.
Molecular genetic variation in this species is relatively
low at mtDNA (Hoelzel et al., 1993; Weber et al. 2000),
allozyme (Bonnell & Selander, 1972; Hoelzel et al., 1993)
MHC (Hoelzel et al., 1999b) microsatellite and minisat-
ellite DNA loci (Hoelzel, 1994; Hoelzel et al., 1999a),
consistent with predictions, given the severity of the Fig. 1 Map illustrating current range of the northern elephant seal,
bottleneck (Hoelzel et al., 1993; Halley & Hoelzel, 1996; and the historical locations of the post-bottleneck population as it
Hoelzel, 1999). As for a number of species (for marine expanded out from the refuge on Guadalupe. Dates indicate first
mammals, see review in Hoelzel et al., 2002), interspe- records of new colonies at specific sites.
cific comparisons suggest the loss of variation caused by a
bottleneck event, however, a direct comparison of the
same population before and after such an event is the Craig, 1979). By the 1880s they were only known to be
only way to fully quantify and assess the impact. Weber at Guadalupe Island and Bahia San Cristobal (on the
et al. (2000) compared mtDNA diversity between modern Baja peninsula), and as far as anyone at the time could
NES samples and five prebottleneck samples, and found determine, all had been killed by 1884 (Scammon, 1870;
novel alleles in the prebottleneck sample (although the Townsend, 1885).
sample size was too small to provide an assessment of The re-growth of the population began on Guadalupe
genetic diversity or statistical comparisons of pre- and with less than 10 seen there in 1892 and 1904, but 40 in
post-bottleneck diversity levels). We expand on these 1907 and 125 in 1911 (Townsend, 1912). By 1918 a
data adding a further 16 prebottleneck samples and four small number of elephant seals were seen on the
new loci (microsatellite DNA loci). relatively nearby Island San Benito, where they began
A key element of this comparison is the assumption breeding in the 1930s. The breeding population on
that the pre- and post-bottleneck samples are from the Guadalupe Island continued to grow to 264 seals in
same source population. The historical data strongly 1922 (Anthony, 1924), and 469 in 1929 (Huey, 1930).
indicate that this is the case. Around the time of the San Miguel Island was colonized in 1925 and the
bottleneck (1860–1890), NES was known only from population began breeding there in the early 1950s. Los
Guadalupe Island, San Benito Island, Cedros Island and Coronados and Santa Barbara islands were colonized in
on the mainland at Bahia San Cristobal (see Fig. 1). 1948, and breeding began in the 1950s (Bartholomew &
These sites are all within a radius of 200 km. There is no Boolootian, 1960; Odell, 1974). An˜ o Nuevo Island
evidence from historical or archaeological data to indicate was colonized in 1955, and the, population, began,
that NES was ever found further south than Baja breeding, in, 1961 (Radford, et, al., 1965).
California, although archaeological data suggest that By 1960, the total population was estimated to be
they were in California 15 000 years ago (Walker & 15 000 with 91% on Guadalupe Island, 8% at San

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Benito Island, and 1% on other islands further north
(Bartholomew & Hubbs, 1960). The expansion continued
onto the Farallones in the 1970s (Le Boeuf et al., 1974).
The clear impression is of a remnant population on
Guadalupe Island that then expanded over the last
110 years into the current species range, with sites near
Guadalupe being colonized first (Fig. 1). All of the
prebottleneck samples used in our study came from
Guadalupe or sites relatively nearby (within approxi-
mately 200 km). Our post-bottleneck sample includes
animals from An˜ o Nuevo and the Channel
Islands (Hoelzel et al., 1993; Weber et al. 2000). Data on
post- bottleneck patterns of dispersal demonstrate that
there is frequent movement between modern rookeries
(about 30% dispersal per annum based on tag recoveries),
and that females breed at newly adopted rookeries (see Le
Boeuf et al., 1974; Le Boeuf, 1981). A comparison
between samples from 109 modern animals from south-
ern California (Channel Islands; Weber et al., 2000) and
40 modern animals from An˜ o Nuevo in central
California (Hoelzel et al., 1993) showed almost identical
mtDNA allele frequencies for the two haplotypes (same
most frequent allele at 71.6 and 72.5%, respectively),
suggest- ing no genetic structure over this geographical
range.
Methods
Genetics
Mitochondrial DNA was amplified from 11 nineteenth
century skulls (Smithsonian collection numbers 38 234,
21 887, 20 927, 38 208, 21 889, 38 285, 21 891, 14 929,
21 738, 21 890, 21 896), of which eight were collected
between 1883 and 1884, three in 1892. DNA was also
extracted from five bone samples from midden sites on
San Miguel Island, dating from the fifteenth to seven-
teenth centuries (Walker et al., 2000). All modern sam-
ples were collected in the 1990s from both the southern
and northern regions of the present-day range. DNA
from modern samples was extracted from skin using
standard protocols (see Hoelzel et al., 1993 for details of
collection and extraction methods). Thirty post-bottle-
neck samples were sequenced for the HVR1 segment of
the control region and pooled with published samples for
a total of 185. Prebottleneck samples were washed in
0.5 M EDTA for 2 h. The clean bone was then wrapped in
foil and crushed. Samples were digested over 24 h at
60 °C in buffer (10 mM Tris, 2 mM EDTA, 10 mM NaCl,
1% SDS, 10 mg mL)1 DTT, 1 mg mL)1 Proteinase K) and
extracted with phenol (2X) and chloroform. Final sup-
ernatent was cleaned on centrocon-30 columns
(Amicon, Beverly, MA, USA). Both water and extract (all
reagents without sample) controls were run. All DNA
extraction and PCR preparation was undertaken in an
isolated ‘ancient DNA’ lab. PCR reactions were set up in
a laminar flow hood, plasticware was UV treated and
double autoclaved, and surfaces were cleaned with
bleach. PCR
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Impact of elephant seal bottleneck 569
amplification of the control region locus was either for
300 bp of the HRV1 region as described in Hoelzel et al.
(1993), or for a 116-bp segment designed around the
variable sites, using the primers: sense- GCCCTATGTA-
TATCGTGC; antisense- GGCTCGTTGGACTAGGTG in
assay conditions: 2 mM MgCl2, 2 mM NTP, 2.4 mg mL)1
BSA, 1 lM each primer, 1 U Taq Gold (hot start); cycled
45 times at 92 °C ) 40 s, 50 °C ) 1 min, 72 °C ) 1 min,
and product sequenced on ABI automated system.
Microsatellite DNA loci NESM11a, cloned from an NES
genomic library (Hoelzel et al., 1999a) and Hg4.2, Hg8.9
and Hg8.10 cloned from a Halichoerus grypus library (Allen
et al., 1995) were amplified from 100 post-bottleneck
NES, and 4–14 nineteenth century NES samples. Locus
NESM11a used primers: sense – TACATTCACAAGG-
CTCAA; antisense – TGTTTCCCAGTTTTACCA and all
microsatellite loci used the same assay conditions and
an annealing temperature of 50 °C (Hg primers as given
in Allen et al., 1995). Amplifications were attempted
from nineteenth century samples at least six times per
locus.
Compliance with the expectations of the Hardy–
Weinberg equilibrium and allele frequency differences
were tested using Fisher exact tests, and the probability
estimated using a Markov chain permutation analyses.
Haplotypic diversities were compared using Welch’s
approximate t-test, which corrects for unequal sample
size and variance (Welch, 1938; Sokal & Rohlf, 1995).
Average heterozygosity was compared using the Mann–
Whitney U-test. Populations that experienced a recent
reduction in effective population size will show a
correlated reduction in allele number and gene
diversity. However, allele number reduces faster than
gene diver- sity, so in a recently bottlenecked
population the gene diversity will be higher than
expected at equilibrium. The program BOTTLENECK
(Cornuet & Luikart, 1996) was used to test for evidence
of a recent bottleneck (using the microsatellite DNA data)
on the basis of this theoretical expectation. The two-
phase model was applied (with 70% stepwise), as this
has been shown to be the most appropriate for
microsatellite DNA data (DiRienzo et al., 1994).
Demographic simulation analyses were after Hoelzel et
al. (1993), and used the same age-specific reproduction
and mortality data.
Morphometrics
The symmetry of bilateral characters was tested using
four measurements from the mandibular region of 11
prebottleneck NES skulls, 43 post-bottleneck NES skulls
and 32 SES skulls (Fig. 1). The absolute value of left
minus right side was used as a measure of the magnitude
of asymmetry (index 1 in Palmer & Strobeck, 1986). All
skulls were measured by the same person (ARH) using a
precision dial caliper. Only female skulls were used as a
result of sexual dimorphism and the paucity of male
skulls. Measurements indicated in Fig. 1 are as follows:
mandibular height (MH); mandibular length (ML);
570 A. RUS HOELZEL ET AL.
mandibular tooth row (MTR) and upper tooth row
(UTR). Measurements were repeated, but for the level
of resolution chosen (±1 mm for ML, ±0.1 mm for MH,
UTR and MTR) no measurement error was detected.
Tests for bias between right and left sides used the
Wilcoxon statistic, and a one-sample t-test against a
mean of zero. Soule´ (1967) points out that ‘asymmetry
is fluctuating if the signed differences between paired
structures are normally distributed with a mean of
zero’. Distributions were tested for normality using the
Kolmogorov–Smirnov test. The Mann–Whitney U statistic
was used to test for the difference between the magni-
tude of asymmetry as measured, for each skull, by the
absolute value of left minus right sides. We also used
linear regressions of left vs. right sides to assess asym-
metry, comparing residual means squared among
regres- sions using an F-statistic. Data were compared
with
Fig. 2 An illustration of measurements taken (a) and linear regressions of left against right MTR for SES (b), prebottleneck NES (c) and post-
bottleneck NES (d).
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and without log transformation, and showed the same
pattern of statistical significance either way (Fig. 2).
Results
Genetic variation
There were only two haplotypes among the 185 post-
bottleneck sequences for 300 bp from the 5¢ mtDNA
control region (115 from Weber et al. 2000; 40 from
Hoelzel et al., 1993; 30 from this study), and seven
haplotypes among 22 samples sequenced for 116 bp
[including two novel haplotypes from six samples (five of
which are prebottleneck) from Weber et al. 2000
sequenced for 179 bp incorporating the 116 bp sequence]
collected in the nineteenth century or before (Table 1).
All haplotypes from nineteenth and twentieth century
 Impact of elephant seal bottleneck 571

Table 1 Haplotype frequency at the control region locus and allele frequency at the microsatellite DNA loci. The nineteenth century sample
includes all samples collected in 1892 or before. Five of the nineteenth century, one of the pre-eighteenth century, and 115 of the modern
control region haplotypes are from Weber et al. (2000).

Haplotype
Control region GTA GAA GAG AAG AAA GAAGC GAGGT

Modern 50 0 0 135 0 0 0
Nineteenth century 7 5 2 1 1 0 0
Pre-eighteenth century 1 2 1 0 0 1 1
Allele

NESM11a 141 143 145 149 151 Hobs Hexp

Modern 0 0 155 0 45 0.38 0.35
Nineteenth century 4 2 18 4 0 0.29 0.54
Hg4.2 137 139 146
Modern 106 0 94 0.44 0.50
Nineteenth century 6 2 8 0.63 0.59
Hg8.9 179 181 183 185 187 193
Modern 0 107 17 0 75 1 0.64 0.57
Nineteenth century 1 1 1 1 6 0 0.60 0.60
Hg8.10 179 181 183
Modern 0 162 38 0.32 0.31
Nineteenth century 1 5 2 0.75 0.53

samples could be defined by the nucleotides at three
positions (196, 214 and 224 from Hoelzel et al., 1993).
Two further variable sites were found only in the midden
samples (positions 225 and 235 from Hoelzel et al., 1993).
All four skulls from 1892 (including one from Weber
et al. 2000 and one analysed in both studies) had the
haplotype GTA (one of two modern sample haplotypes).
The AAG haplotype shows a significantly different
frequency in the post-bottleneck (lower 95% confidence
estimate ¼68.5%) compared with the prebottleneck
sample (upper 95% confidence estimate ¼ 13.3%).
Haplotypic diversity (Nei & Tajima, 1981) was signifi-
cantly greater in the prebottleneck sample (from 1884
and before; h 0.804¼ ± SE 0.069) than in the post-
bottleneck sample (h 0.41
¼ ± 0.028; t¢ 5.28,
¼ P<
0.001). This difference is still significant when the 1892
samples are counted among the prebottleneck samples (t¢
¼ 5.69, P < 0.001). An exact test comparing
the haplotype frequencies shown in Table 1 was also
highly significant (P < 0.0001).
In an earlier study, the size of the putative bottleneck
had been back-calculated using a monte carlo
simulation model (500 reiterations with random seeds;
Hoelzel et al., 1993), based on life history data collected
over 25 years for NES, and an estimate of prebottleneck
genetic variation based on the congeneric SES. We
repeated this analysis using all the same parameters as
in Hoelzel et al. (1993), but using the estimate of
prebottleneck mtDNA diversity from the data presented
in Table 1. The model bottleneck event is imposed
instantaneously, and the prebottleneck parameters are
based on a long-term study of NES (see Le Boeuf &
Reiter, 1988). A census in 1960 provided a population
estimate of 15 000 seals at that time, and the modern
level of genetic diversity at this locus is two haplotypes
(Table 1). Therefore, the simu- lation is run for 76 years,
and the number of simulations

Table 2 For putative bottlenecks of 5–20 seals, results are presented for the number of simulation runs that resulted in 15 000 seals and
two haplotypes at the end of the run (outcome match), the number of runs that did not go to extinction (out of 500), and the mean final
population size and haplotype number at the end of the run. Details for the most likely event are shown in bold.

Bottleneck size Outcome match Surviving runs End population size End Haplotype
(SD) number (SD)

5 3 170 5367 (5597) 1.33 (0.48)

[2] [128] [5354 (5907)] [1.41 (0.58)]
10 13 423 15 195 (12 067) 1.97 (0.72)
[8] [408] [14 963 (11 423)] [2.44 (1.09)]
15 6 493 27 277 (16 281) 3.18 (1.08)
[4] [485] [27 867 (16 139)] [3.88 (1.54)]
20 2 500 40 386 (19 493) 4.17 (1.14)
[2] [499] [41 040 (19 643)] [5.34 (1.83)]

Values in brackets are from Hoelzel et al. (1993).

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572 A. RUS HOELZEL ET AL.

Table 3 Fluctuating asymmetry of cranial measures comparing prebottleneck NES (bBN), post-bottleneck NES (aBN) and SES skulls.

Range of ŒL–R Œ bBN vs. aBN SES vs. aBN SES vs. bBN

Measure bBN aBN SES z F z F z F

UTR 0– 0.03 0 0.79 1.75 4.97 8.78 1.81 5.02

0.3 )0.62 )0.23 *** *** **
MTR 0 0.04– 0– 3.09 6.73 5.23 8.93 0.50 1.32
)0.54 1.77 0.49 ** *** *** ***
ML 0 0–0.5 0– 0.63 1.63 2.25 1.47 2.08 1.11
)0.5 1.10
MH 0– 0–0.4 0– 0.21 1.53 2.78 3.23 2.03 4.94
0.5 0.28 ** *** **

**P < 0.005, ***P < 0.0001.

Data for the range of absolute values of left–right sides (ŒL–R Œ) is shown in centimetres without transformation. UTR ¼ upper tooth row,
MTR ¼ mandibular tooth row, ML ¼ length of the mandible, MH ¼ height of the mandible. Mann–Whitney U-test (z) comparing ŒL–R Œ
and an F-statistic (F) comparing RMS are given.

that reach 15 000 (±1000) and two haplotypes simulta-
neously are noted to provide a probability distribution for
different putative bottleneck sizes (Table 2). Although
the standard deviations are high for population size and
haplotype number estimates (due to demographic sto-
chasticity), the difference in the genetic and demographic
distributions permits some resolution (see Hoelzel et al.,
1993 for further discussion). The refined estimate pre-
sented here is consistent with the published estimate
(Table 2). Both analyses suggest a bottleneck of less
than 20 seals with a bottleneck size of 10 seals showing
the highest probability.
For the microsatellite DNA loci, there were between
one and three alleles per locus (total of seven for all
four loci combined) unique to the nineteenth century
sample set (Table 1). One allele found in the post-
bottleneck sample at Hg8.9 in just one heterozygous
individual, and not in the nineteenth century sample,
could be a new mutation. The observed heterozygosity
was not signifi- cantly different from that expected
under the Hardy– Weinberg equilibrium for any of the
loci. Even so, there were few enough heterozygotes in
the nineteenth cen- tury sample at NESM11a to suggest
possible allelic dropout (despite repeat amplifications).
Alleles apparently unique to the nineteenth century
samples would need to be present at a frequency of less
than 1.4% for the mtDNA locus, and 1.3% for the
microsatellite loci to have gone undetected given the
post-bottleneck sample sizes (based on a 95% confidence
estimate for a Poisson probability of zero). While the
prebottleneck sample size for microsat- ellite loci was
too small to provide realistic estimates of allele
frequencies, given the diversity seen in this small
sample, it is probable that a larger sample size would
reveal further allelic diversity. Exact tests comparing the
allele frequencies shown in Table 1 were highly signifi-
cant for all loci (P < 0.0001). Average He was greater in
the nineteenth century sample (0.565 vs. 0.433),
although the difference was not significant at the 95%
confidence level (z ¼ )1.73, P ¼ 0.08). The microsat-
ellite DNA data also enable us to test for the signature of a
recent bottleneck based on the comparison of observed
gene diversity with that expected at equilibrium (Cornuet
& Luikart, 1996), although the power of the test is low
with only four loci. Both the sign test (P¼ 0.04) and
Wilcoxon test (P ¼ 0.03) show a significant excess for all
loci, indicative of a recent bottleneck.
Morphometric analyses
Measures of ŒL–R Œ are given for all comparisons
in Table 3. The distributions of L–R for all NES measure-
ments were not significantly divergent from normal
distributions, and the means for NES and SES distribu-
tions were not significantly different from zero, indica-
ting that the observed asymmetry is FA. Some SES
distributions were leptokurtic, but this does not imply
antisymmetry, and therefore does not alter interpreta-
tions regarding FA. One measurement (MTR) showed
significantly greater FA in post compared with prebot-
tleneck NES samples, and three of the four measures
(including MTR) showed significantly greater FA in post-
bottleneck NES compared with SES samples (Table 3).
There was no significant difference between the prebot-
tleneck NES and SES samples at MTR, or for any other
characters based on the non-parametric tests (see
Table 3). The comparison for MTR is illustrated in
Fig. 2 using linear regressions of right vs. left sides. The
robustness of the effect for MTR was tested by comparing
equal sized sample sets for pre- and post-bottleneck skulls
(every fourth sample was included for the post-bottle-
neck sample-set three times for three independent tests),
and the post-bottleneck increase in FA was still significant
(z ¼ 2–3, P ¼ 0.015–0.009).
Discussion
These data show a strong increase in FA for one
morphometric measure (MTR) in post-bottleneck NES

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compared with either prebottleneck NES or SES, and the
direct loss of genotypes at all five loci investigated. The
direct comparison of historical samples indicates that this
is the result of a bottleneck that most probably occurred
when over 400 seals were taken from an already depleted
population between 1880 and 1884. Although the
pattern of allele frequencies in the prebottleneck sample
cannot be accurately assessed at all loci because of small
sample sizes, the striking difference at some loci in the
pre- and post-bottleneck samples reflects expectations
about the stochastic disruption of allele frequencies
caused by sampling effects following bottleneck events
(cf. Luikart et al., 1998). The absence in the prebottleneck
sample of a post-bottleneck allele at one of the five loci
(NESM11a) most probably reflects this type of allele
frequency distortion. An incomplete representation of
alleles as a result of prebottleneck sample size is also a
factor. For example, for a sample size of 14 (as for
NESM11a) the allele would need to be present at 20% to
give 95% confidence of detection (based on a binomial
distribution).
The direct loss of alleles and the significant change in
measures of diversity are the primary result from the
molecular genetic data. However, tests against equilib-
rium expectations are also consistent with the bottleneck
interpretation, although a larger number of loci should
be included for the BOTTLENECK program for a more
thorough test. Earlier work based on simulation models
showed the consistency of molecular data for the modern
population with the bottleneck interpretation (Hoelzel
et al., 1993; Hoelzel, 1999). Using the prebottleneck
mtDNA data from this study instead of the estimate
based on SES (as presented in Hoelzel et al., 1993)
reinforced the earlier interpretation. In Hoelzel et al.
(1993) the most probable bottleneck according to the
simulation was smaller that 20 individuals, and this was
still true using the actual prebottleneck data.
Heritability of FA is typically low (e.g. Leamy, 1999),
and we do not have the necessary data to assess the
genetic component of the effect observed. Further,
because of incomplete knowledge about the independ-
ence of the measured characters and the developmental
processes involved, it is not clear that this result implies a
generalized decrease in canalization, as only one charac-
ter shows a strong and significant increase in FA.
However, the pattern of post-bottleneck asymmetry seen
in our results is consistent with results for some
other populations thought to have undergone severe
bottlenecks (Kieser & Groeneveld, 1991; Hutchison &
Cheverud, 1995). Further, the magnitude of FA in MTR
is comparable with that seen for some other inbred
populations, such as FA based on cranial measures in the
dama gazelle (Alados et al., 1995).
Swaddle et al. (1994) describe two potential problems
(relevant to this study) with using museum material for
studies of FA. First, human collections for museums
may be biased towards symmetrical and otherwise
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Impact of elephant seal bottleneck 573
‘aesthetic’ samples. In this case, all skulls were from
museums (so any bias should be similar for different
samples) and most were collected opportunistically.
Secondly, wear and damage asymmetry may not be
discernible from FA. However, any wear would tend to
diminish an apparent effect of increased post-
bottleneck asymmetry, because the greater wear would
be on the older (prebottleneck) skulls. Furthermore,
either of these factors should affect the recent NES and
SES skulls in a similar way. A greater difficulty is the
small sample size for prebottleneck skulls. However, the
post- bottleneck effect was strong enough that sub-
samples of
11 skulls from the larger NES post-bottleneck sample
still showed a significant increase in asymmetry for MTR
compared with the prebottleneck NES sample. Sampling
biases based on kinship are unlikely as samples were
collected over a period of years.
These results show that the NES bottleneck had a
quantifiable impact on genetic diversity and FA in the post-
bottleneck population. An understanding of the
details of such impacts is important for understanding the
role of demographic fluctuations on the evolution and
loss of diversity.
Acknowledgments
We thank Charles Potter, John Heyning, Philip Unitt,
Maria Rutzmoser, Robert DeLong, Phil Walker, Doug
Kennett, Doug Jones, the Smithsonian Institution, the
Los Angeles County Museum, the London Museum of
Natural History, the San Diego Natural History Museum,
and at the Harvard University Museum of Comparative
Zoology for material and assistance.
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Received 8 February 2002; revised 15 March 2002; Accepted 22 March
2002