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DOI 10.1007/s00227-011-1828-y
ORIGINAL PAPER
Received: 29 June 2011 / Accepted: 19 October 2011 / Published online: 5 November 2011
Springer-Verlag 2011
Communicated by M. I. Taylor.
average number of alleles (NA = 9) and observed heterozygosity (HO = 0.58) both low. The low levels of diversity
seen in both the mtDNA and the microsatellite may be due
to historical sea level fluctuations and concomitant loss of
shallow water habitat. Eight of the 10 pair-wise western
Atlantic FST estimates for mtDNA indicated significant
genetic subdivision. Pair-wise FST values for the microsatellite loci indicated a similar pattern as the mtDNA. The
western Atlantic population of nurse sharks is genetically
subdivided with the strongest separation seen between the
offshore islands and mainland Brazil, likely due to deep
water acting as a barrier to dispersal. The eastern and
western Atlantic populations were closely related. The
eastern Pacific individual is quite different from Atlantic
individuals and may be a cryptic, sister species.
Introduction
S. A. Karl A. L. F. Castro
Department of Biology, University of South Florida, SCA110,
4202 E. Fowler Ave., Tampa, FL 33620, USA
Present Address:
S. A. Karl (&)
Hawaii Institute of Marine Biology, University of Hawaii,
PO Box 1346, Manoa, Kaneohe, HI 96744, USA
e-mail: skarl@hawaii.edu
Present Address:
A. L. F. Castro
Departamento de Zootecnia, Universidade Federal de Sao Joao
del Rei, Av. Visconde do Rio Preto, S/N, Sao Joao Del Rei,
MG 36301-360, Brazil
R. C. Garla
Departamento de Botanica, Ecologia e Zoologia, Centro de
Biociencias, Universidade Federal do Rio Grande do Norte,
Campus Universitario, Lagoa Nova, Natal, RN 59072, Brazil
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Results
We sequenced a 1,166 nt fragment of mtDNA containing
the 30 end of the cyt b gene, both tRNAPro and tRNAThr and
the 50 end of the control region from a total of 146 nurse
sharks. On average, this fragment was of 13.1% guanine,
33.9% adenine, 31.3% thymine, and 21.6% cytosine. There
were 34 polymorphic sites consisting of 32 substitutions
(25 transitions and seven transversions) and two indels,
resolving 12 haplotypes (Table 1; Fig. 2; Table S1; GenBank Accession numbers JF950288-JF950299). The vast
majority of the polymorphism was found between the
Pacific (haplotype H12) and Atlantic haplotypes with 19
substitutions (14 transitions and five transversions) and two
indels. The overall haplotype (h) and nucleotide diversities
(p) for the mtDNA of the western Atlantic populations
were 43 5% (standard deviation) and 0.04 0.04%,
respectively (Table 2). Considering all samples (i.e.,
including the eastern Atlantic and Pacific), h and p were
slightly larger (Table 2). H1 was the most prevalent
haplotype, found in all western Atlantic locations and in
493
BRI
37
0.43 0.06
0.0004 0.0004
NTL
15
0.56 0.09
0.0005 0.0005
BMN
FLD
21
37
6
4
0.43 0.13
0.16 0.08
0.0004 0.0004
0.0001 0.0002
Haplotype
BLZ
Geographic location
BRI
NTL
BMN
FLD
BLZ
AFR
PNM
Total
H1
26
16
34
20
105
H2
13
19
H3
H4
11
11
2
H5
H6
H7
H8
H9
H10
H11
H12
Total
37
15
21
37
33
146
33
0.49 0.04
0.0004 0.0004
Western Atlantic
143
10
0.43 0.05
0.0004 0.0004
Total
146
12
0.48 0.05
0.0008 0.0006
Note that overall values are presented for the western Atlantic and for
the entire sample set (i.e., including samples from East Pacific and
East Atlantic). Locations abbreviations are as in Fig. 1
BRI
BRI
NTL
0.275
BMN
FLD
0.157
a
BLZ
0.218
0.334
NTL
0.012
0.109
0.266
-0.034
BMN
FLD
0.020
0.016
0.030
0.011
-0.003
0.003
0.185
0.318
BLZ
0.009
0.005
0.006
-0.004
P = 0.06
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transversion) when compared to the western Atlantic haplotypes. The K2P distance estimates indicate an average of
1.6% divergence between the Pacific and the western
Atlantic haplotypes. This is much smaller than the divergence found among the four species in the Genus Chiloscyllium (7.59.8%), but similar to the divergence between
the two species of Orectolobus (1.8%, Fig. 3). Interestingly, the Orectolobus spp. are thought to have formed *2
million years ago (Corrigan and Beheregaray 2009), which
is fairly close to the estimated age of the closing of the
Isthmus of Panama (*3.5 million years ago; Coates et al.
1992) that would have divided the Pacific and western
Atlantic nurse shark populations.
Discussion
Genetic diversity
Nurse sharks are reported to be the most abundant shark
species in shallow, coastal waters (Castro 2000; Hazin
et al. 2000; Castro and Rosa 2005; Heithaus et al. 2007).
Precise estimates of population sizes are mostly lacking,
but in the Atol das Rocas Marine Reserve, Brazil, there are
at least 400 individuals living in one *6 km2 area (Castro
and Rosa 2005). Even so, nurse sharks exhibit the fourth
lowest overall mtDNA haplotype (h = 43%) and lowest
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