Phylogeography and population genetic structure of an exotic invasive brine shrimp, Artemia Leach, 1819 (Crustacea : Anostraca), in Australia

Abstract

Native American Artemia franciscana has become an introduced species in the Old World due to the rapid development of the aquaculture industry in Eurasia. The recent colonisation of A. franciscana in Mediterranean regions and Asia has been well documented, but Australia is a continent where the dispersal of this species is not well understood. In the present study, we sequenced the cytochrome oxidase subunit I (COI) and examined the phylogenetic relationships, haplotype network and population genetic structure of Artemia from four geographical localities in Australia and two American native localities. Our results confirmed the colonisation of Australia in all four localities by A. franciscana. First, we document the occurrence of Artemia in Mulgundawa and St Kilda localities in Australia. The Dampier population is a monomorphic population, but there is high genetic variation and a degree of demographic expansion observed in other introduced A. franciscana populations in Australia. This observation suggests an interaction between environmental conditions and adaptive potentials of A. franciscana. Our findings imply that populations from St Kilda and Port Hedland might have originated from a San Francisco Bay source, while the two other locations resulted from admixture between Great Salt Lake and San Francisco Bay sources, perhaps resulting from secondary introduction events.

Full text

Phylogeography and population genetic structure of an
exotic invasive brine shrimp, Artemia Leach, 1819
(Crustacea : Anostraca), in Australia
Alireza Asem
A,D,E
, Amin Eimanifar
B,D,E
, Weidong Li
A
, Pei-Zheng Wang
A
,
Samantha A. Brooks
C
and Michael Wink
B
A
College of Life Sciences and Ecology, Hainan Tropical Ocean University, Yucai Road, Sanya 572000, China.
B
Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Im Neuenheimer Feld 364,
69120 Heidelberg, Germany.
C
Department of Animal Sciences, University of Florida, Gainesville, FL 32611, USA.
D
These two authors contributed equally to this paper.
E
Corresponding authors. Email: asem.alireza@gmail.com; amineimanifar1979@gmail.com
Abstract. Native American Artemia franciscana has become an introduced species in the Old World due to the rapid
development of the aquaculture industry in Eurasia. The recent colonisation of A. franciscana in Mediterranean regions and
Asia has been well documented, but Australia is a continent where the dispersal of this species is not well understood. In the
present study, we sequenced the cytochrome oxidase subunit I (COI) and examined the phylogenetic relationships,
haplotype network and population genetic structure of Artemia from four geographical localities in Australia and two
American native localities. Our results confirmed the colonisation of Australia in all four localities by A. franciscana. First,
we document the occurrence of Artemia in Mulgundawa and St Kilda localities in Australia. The Dampier population is a
monomorphic population, but there is high genetic variation and a degree of demographic expansion observed in other
introduced A. franciscana populations in Australia. This observation suggests an interaction between environmental
conditions and adaptive potentials of A. franciscana. Our findings imply that populations from St Kilda and Port Hedland
might have originated from a San Francisco Bay source, while the two other locations resulted from admixture between
Great Salt Lake and San Francisco Bay sources, perhaps resulting from secondary introduction events.
Additional keywords: Australian Artemia, biodiversity, introduced species, mtDNA-COI.
Received 23 November 2018, accepted 3 April 2019, published online 26 April 2019
Introduction
The brine shrimp Artemia is a primitive microcrustacean
inhabiting many hypersaline habitats worldwide such as inland
salt lakes, coastal saltworks, salt ponds and lagoons (Van
Stappen 2002). Artemia can withstand extreme environmental
conditions such as high salinity (7.0 –340 g/L) and ionic
compositions in the natural environment due to its unique
osmoregulation mechanism (Post and Youssef 1977; Bowen
et al.1985; Lenz 1987; Browne et al.1988; Liu and Zheng 1990).
The genus Artemia consists of seven bisexual species and
numerous parthenogenetic populations with different ploidy
levels (Asem et al.2010; Asem et al.2016). Three bisexual
species occur naturally in the New World: Artemia monica
Verrill, 1869 (Mono Lake, USA), Artemia franciscana Kellogg,
1906 (North America, Central America and South America) and
Artemia persimilis Piccinelli & Prosdocimi, 1968 (Argentina and
Chile). The other four bisexual species are native to the Old
World, namely Artemia salina (Linnaeus, 1758) (Mediterranean
basin), Artemia urmiana Gunther, 1899 (Lake Urmia, Iran, and
the Crimean salt lakes, Russia), Artemia sinica Cai, 1989 (China
and Mongolia) and Artemia tibetiana Abatzopoulos, Zhang &
Sorgeloos 1998 (Qinghai–Tibetan Plateau, China). Previous
studies have documented that the Tibetan populations were
placed in two different groups in the phylogenetic trees of the
mitochondrial COI marker, while all of them represented a single
clade when analysed with the nuclear marker ITS1 (Maccari et al.
2013; Eimanifar et al.2014). This contradiction could be
attributed to a hybridisation event that occurred between two
ancestors. Two new species were recently described from
Mongolia (Asia), namely Artemia frameshifta and Artemia
murae (Naganawa and Mura, 2017), although the taxonomic
status of these taxa requires confirmation. The biosystematics
of these two new species were determined by single individual
differentiation from the cytochrome oxidase subunit I (COI)
sequence combined with morphological parameters, whereas
morphometric study and population genetic analysis have
Journal compilation CSIRO 2018 www.publish.csiro.au/journals/ajz
CSIRO PUBLISHING
Australian Journal of Zoology, 2018, 66, 307–316
https://doi.org/10.1071/ZO18077

not previously been examined. Additionally, the existence
of males has not been investigated in A. frameshifta so that
the reproductive mode of that population is still doubtful.
Parthenogenetic populations are obligate clones containing di-,
tri-, tetra-, penta- and heteroploids, and inhabit the Old World and
Oceania (Sun et al.1999; Abatzopoulos et al. 2003).
Artemia has been broadly used as live food in the fishery and
aquaculture industry, especially in the coastal areas of eastern
Asia (Van Stappen 2008). It is also used to improve the quality of
sodium chloride production in solar salt-fields by aiding in the
control of phytoplankton blooms and increasing the density of
red-pigmented bacteria to accelerate evaporation (Jones et al.
1981; Ruebhart et al.2008). Artemia is a model organism in many
biological fields, including phylogeography and population
genetics (Kappas et al.2011), molecular and cellular biology (Li
et al.2017), bioassay toxicity (Rajabi et al.2015) and
bioencapsulation (Vázquez-Silva et al.2017).
Since 1950, cysts of A. franciscana have been exported
overseas from the USA for applications in fishery markets.
Genetic studies have documented that these exports originated
primarily from two major natural sources in the USA, namely the
Great Salt Lake, Utah (GSL), and San Francisco Bay, California
(SFB) (Van Stappen 2008; Muñoz 2009; Eimanifar et al.2014).
Phylogeographic analysis revealed that the expansion of
A. franciscana to non-native regions has resulted in rapid
colonisation of numerous regions across Eurasia (Amat et al.
2005; Mura et al.2006; Van Stappen 2008; Muñoz 2009; Ben
Naceur et al.2010; Scalone and Rabet 2013; Eimanifar et al.
2014; Horvath et al.2018). Phylogenetic analysis of Artemia has
not previously been conducted in Australia because of difficulties
in obtaining adequate samples.
Previous studies have suggested the introduction of
A. franciscana into Australia (Clark and Bowen 1976; Geddes
1979,1981; Abreu-Grobois and Beardmore 1982; Geddes and
Williams 1987; Vanhaecke et al.1987; Pinder et al.2002;
McMaster et al.2007) but there was no evidence using genetic
barcoding to support the regional colonisation of A. franciscana
in Australia. The aim of the present study was to perform a
phylogenetic analysis of Artemia populations from Dampier,
Mulgundawa, Port Hedland and St Kilda in Australia to confirm
the taxonomical status of Artemia in these localities. Here we
sequenced the mitochondrial COI gene and determined the
genetic diversity, population genetic structure and the genetic
source of Artemia populations as compared with two American
native populations of A. franciscana from GSL and SFB.
Materials and methods
Origin of samples and sample analysis
In total, 67 individuals of bisexual Artemia were collected from
four geographical sites in Australia in summer 2011 (Fig. 1).
The sampling sites, with their abbreviations, geographical
coordinates, IPMB code numbers and number of individuals
analysed, are summarised in Table 1.
Total DNA was separately extracted from part of the antenna
of male and female shrimps (1 : 1) following the Chelex
®
100
Resin method (Bio-Rad Laboratories, USA). The samples were
crushed, incubated for 2.5–3hat60
C (tubes were vortexed
every 30 min) and then a final 10 min at 80C. Then the tubes were
centrifuged at 10 000 rpm for 1 min and the supernatant phase
was directly used in the PCR reaction (Montero-Pau et al.2008;
Eimanifar and Wink 2013; Asem et al.2016). All extracted DNA
was stored at –80C for further genetic analyses.
A fragment of the mitochondrial cytochrome oxidase subunit I
(COI) was amplified. PCR was performed in a final reaction
volume of 50 mL in a thermocycler (Biometra, Tgradient,
Germany) with Taq DNA polymerase (Bioron, GmbH, Germany)
according to conditions published previously (Eimanifar and
Wink 2013). The COI partial fragment (~588 bp) was amplified
using the metazoan invertebrates’universal primers LCOI490/
HC02198 (Folmeret al.1994). PCR amplification was carried out
under the following conditions: a cycle of 3 min at 94C, followed
by 35 cycles of 45 s at 94C, 60 s at 45C, and 60 s at 72C, with a
final step of 5 min at 72C. Before sequencing, PCR products were
purified using standard procedures (Eimanifar and Wink 2013).
Sequence alignment and phylogenetic analyses
Sequences were aligned using MEGA 7.0.26 with default
parameters (Kumar et al. 2016). A lack of pseudogenes enabled
utilisation of the protein-coding sequence; additionally, multiple
mutations or deletion(s) and duplication(s) were not observed.
To estimate the phylogenetic relationship among samples
collected from Australia and other species, COI reference
sequences of bisexual species and parthenogenetic populations
including di-, tri-, tetra- and pentaploidy were downloaded from
GenBank (Table 2). The phylogenetic tree was generated using
Bayesian Inference (BI) (Huelsenbeck and Ronquist 2001), as
implemented in MrBayes 3.2.2 on XSEDE (Miller et al.2010).
For BI the best-fitting nucleotide substitution model was
calculated based on MrModelltest 2.2 (Nylander 2004) and HKY
+G was chosen as the best-fit model. Additionally, for posterior
probabilities, the values <0.94 and 0.95 were considered to be
low and high, respectively (Alfaro et al.2003).
To find the origin and genealogical relationships among
haplotypes of Australian samples and A. franciscana (more
information in Results), a median network was performed using
the median-joining algorithm in Network 5.0.0.3 (Bandelt et al.
1999). The sequences of A. franciscana were chosen from
two natural habitats in the USA: GSL and SFB (Table S1,
Supplementary Material).
For each population,the number of polymorphic sites (S), total
number of mutations (Eta), number of haplotypes (h), haplotype
diversity (Hd), haplotype ratio (Hr), nucleotide diversity (Pi) and
average number of nucleotide differences (k) were calculated
using DnaSP 5.10 (Librado and Rozas 2009). Expected
heterozygosity, F
ST
(an overall population differentiation index),
mismatch distribution, Harpending’s Raggedness index (Hri) and
sum of squared deviations (SSD) were computed in Arlequin 3.5
(Excoffier and Lischer 2010).
Results
All COI sequences of A. franciscana from Australia had 18
variable sites, of which four sites were parsimony informative
and 14 sites were singletons. The total COI sequences of native
American A. franciscana displayed 12 variable sites, of which
five sites were parsimony informative and seven sites were
singletons.
308 Australian Journal of Zoology A. Asem et al.

(a)
(b)(c)
(d)(e)
Fig. 1. (a) Map of Artemia sampling sites in Australia, (b) DAM: Dampier, (c) HED: Port Hedland, (d) SKI:
St Kilda, (e) MUL: Mulgundawa. Map data Google 2018.
Phylogeography of invasive Artemia in Australia Australian Journal of Zoology 309

The phylogenetic tree revealed that all examined Artemia
individuals from Australia clustered in the clade of
A. franciscana (Fig. 2). The haplotype distribution network
analysis of the A. franciscana complex was performed to
determine the population structure of individuals, but a
geographically unique haplotype could not be distinguished
(Fig. 3). Most sequences belonged to H1 (31.3%) and H2
(28.5%). The haplotype frequency of A. franciscana from SFB
for H1, H2 and H3 haplotypes were 70.3%, 16.2% and 10.8%,
respectively. A. franciscana from GSL grouped in the H2 and H4
haplotypes with frequencies of 62.3% and 26.2%, respectively.
Two localities from Australia had the greatest H1 haplotype
frequency: HED (80.96%: 17 individuals out of 21) and SKI
(70%: 7 individuals out of 10). There were seven haplotypes
around H1 with a frequency of 1.0 (Fig. 3). All A. franciscana
sequences from the DAM locality were recovered in H3. The
MUL locality consisted of 73.1% H3 and 11.5% H2 haplotype
frequencies (Fig. 3, Tables S2, S3 in the Supplementary
Material).
The population differentiation test (F
ST
) among the
examined populations suggested that there is no significant
differentiation between DAM and MUL (4.8%), SKI and HED
(1.3%) and SKI and SFB localities (7.8%), respectively.
A significant population differentiation was observed between
the GSL and SFB localities (60.9%). The highest variation was
revealed between DAM and SKI (90.9%) and DAM and HED
(91.9%), and the lowest was observed between HED and SFB
(10.3%), respectively (Table 3).
All estimated genetic indices for the examined localities are
summarised in Table 4. The lowest genetic variation was
observed in the DAM location, which had only a single
haplotype. Two localities, SKI and MUL, exhibited the highest
genetic variation. The highest-ranking levels of Hd (0.533 
0.180) and Hr (0.4) were found in SKI, whereas the MUL locality
had the highest values for Pi (0.00349 0.0023), k (1.554) and
H
exp
(0.0013 0.016) (Table 4). Between the native American
populations, the highest values for Hd (0.550 0.058) and Hr
(0.15) were found in GSL, whereas the other genetic indices were
highest in the SFB locality. In total, among all American and
Australian populations, the highest values of Pi (0.00349 
0.0023), expected heterozygosity (0.0013 0.016) and k (1.554)
were recorded in MUL. The highest values for Hr (0.4) and Hd
(0.550 0.058) were observed in SKI and GSL, respectively
(Table 4).
We calculated mismatch distributions for pairwise
differences from exotic and native populations of A. franciscana.
These revealed that the SKI, HED and GSL localities had a
unimodal pattern, whereas MUL and SFB localities showed a
pattern likely to be multimodal. Additionally, the indices of SSD
and Hri for all examined localities were non-significant, except
Table 1. Origin of Artemia samples used for this study
IPMB, Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Germany; WA, Western Australia; SA, South Australia
Site Abbreviation Geographic coordinates IPMB voucher
no.
No. of
individuals
Reference
Dampier, WA DAM 2042019.3800S, 1164202.0900E 66843 10 Ruebhart et al.(2008); McMaster et al.(2007)
Mulgundawa, SA MUL 3517039.8400S, 13912037.7600 E 66844 26 This study
Port Hedland, WA HED 2020026.5700S, 11839020.9800E 77593 21 Ruebhart et al.(2008)
St Kilda, SA SKI 3444009.4900S, 13832020.3400 E 66849 10 This study
Table 2. Species information and GenBank accession numbers
Pop., population
Species/population Abbreviation No. of individuals Accession nos Reference
A. urmiana URM 4 JX512748–751 Eimanifar and Wink (2013)
A. sinica SIN 4 KF691298–301 Eimanifar et al.(2014)
A. tibetiana TIB 4 KF691215–218 Eimanifar et al.(2014)
A. salina SAL 4 KF691512–515 Eimanifar et al.(2014)
A. persimilis PER 4 DQ119647 Hou et al.(2006)
HM998992 Maniatsi et al.(2011)
EF615594 Wang et al.(2008)
EF615593 Wang et al.(2008)
A. franciscana FRA 4 KJ863440–443 Eimanifar et al.(2014)
Diploid Pop. DI 4 KU183949–952 Asem et al.(2016)
Triploid Pop. TRE 3 HM998997–999 Maniatsi et al.(2011)
Tetraploid Pop. TETR 4 KU183954–957 Asem et al.(2016)
Pentaploid Pop. PEN 4 KU183968–971 Asem et al.(2016)
Unidentified
A
DAM 10 MK613273–282 This study
MUL
B
26 MK613283–308 This study
HED 21 MK613309–329 This study
SKI
B
10 MK613330–339 This study
A
Australian samples
B
New record site.
310 Australian Journal of Zoology A. Asem et al.

for MUL, where we observed a significant SSD value (P<0.001)
(Fig. 4).
Discussion
The occurrence of American A. franciscana in Australia had been
suggested for three geographical regions (11 localities) in
Australia, including western (seven sites), southern (one site) and
Queensland (three sites) (Ruebhart et al.2008). Our findings
confirmed the colonisation of Australia with A. franciscana.
Previously, parthenogenetic populations have been reported
from two localities in Australia –Port Hedland and Dry Creek
(Van Stappen 2002)–but our findings could not support the
existence of parthenogenetic populations in these localities. In
the present study, we have documented the first scientific record
of Artemia in Mulgundawa and St Kilda localities in southern
Australia, which are clearly colonised by A. franciscana. Brine
shrimps from GSL and SFB have often been introduced to other
regions of the world for aquaculture production of Artemia cysts
and biomass (Sorgeloos et al.2001; Amat et al.2005; Eimanifar
et al.2014; Muñoz et al.2014; Saji et al.2019) therefore these
populations were included in this analysis.
Mitochondrial markers reflect the maternal evolutionary
pathway and are important for understanding the passageways
0.04
1
0.99
0.97
0.95
10.96
0.99
1
1
Diploid P.P.
URM
Triploid P.P.
TIB
SIN
Tetraploid P.P.
Pentaploid P.P.
FRA
MU MUDAM HED SKI
Daphnia
SAL
PER
Fig. 2. COI phylogeny of Artemia based on a Bayesian inference approach. The numbers behind major nodes denote posterior
probabilities. Daphnia tenebrosa (HQ972028) was used as an outgroup. Red dots show the position of reference sequences of
A. franciscana. P.P., parthenogenetic population; URM, Artemia urmiana; TIB, Artemia tibetiana; SIN, Artemia sinica; FRA,
Artemia franciscana; PER, Artemia persimilis; SAL, Artemia salina; DAM, Dampier; MUL, Mulgundawa; HED, Port Hedland; SKI,
St Kilda.
Phylogeography of invasive Artemia in Australia Australian Journal of Zoology 311

and elucidating the source of invasive species in non-indigenous
habitats (Ashton et al.2008; Ficetola et al.2008; Mabuchi et al.
2008; Gaubert et al.2009). The genetic structure of
mitochondrial COI in the Mediterranean Artemia populations
have clearly documented an invasion of A. franciscana from the
GSL and SFB (Muñoz et al.2014; Horvath et al.2018).
Phylogeographical analysis of Asian populations has
documented colonisation by A. franciscana from multiple
origins in both America and Europe (Eimanifar et al.2014).
Haplotype distribution revealed A. franciscana in Al Wathba
Wetland Reserve (United Arab Republic; Abu Dhabi), likely
originating from the GSL (Saji et al.2019). Our results strongly
support the idea that A. franciscana found in the HED and SKI
H17
H16
H15
H18
MUL
SKI
DAM
HED
SFB
GSL
H19
H20
H21
H22
H23 H13
H12 H11
H10
H9
H8
H2
H3
H14 H4
H6
H7
[3]
[3]
H1
H5
Fig. 3. The relationship of COI haplotype distribution among Artemia franciscana individuals from Great Salt
Lake (GSL), San Francisco Bay (SFB) and Australian populations (MUL, Mulgundawa; SKI, St Kilda; HED, Port
Hedland; DAM, Dampier).
Table 3. Pairwise population matrix of F
ST
values from COI loci
Results are shown as percentages. ns, non-significant; *, P<0.05; **,
P<0.001
Site DAM MUL ADE HED GSL
MUL 4.8
ns
SKI 90.9** 63.1**
HED 91.9** 68.6** 1.3
ns
GSL 84.1** 68.1** 75.7** 77.5**
SFB 69.7** 52.2** 7.8
ns
10.3** 60.9**
Table 4. Population genetic indices for Australian and native American A. franciscana based on COI loci
N, no. of sequences; S, no. of polymorphic (segregating) sites; Eta, total no. of mutations; h, no. of haplotypes;
Hd, haplotype (gene) diversity; Hr, no. of haplotypes/no. of sequences; Pi, nucleotide diversity; k, average no.
of nucleotide differences; Exp. Het., expected heterozygosity
Genetic indices DAM MUL SKI HED GSL SFB
N102610216137
S0103485
Eta 0 10 3 4 8 5
h164594
Hd
(s.d.)
0 0.465
(0.116)
0.533
(0.180)
0.352
(0.131)
0.550
(0.058)
0.480
(0.087)
Hr 0.1 0.23 0.4 0.24 0.15 0.11
Pi
(s.d.)
0 0.00349
(0.0023)
0.00135
(0.00131)
0.00085
(0.0009)
0.0015
(0.0012)
0.0025
(0.0018)
k 0 1.554 0.600 0.381 0.666 1.135
Exp. Het.
(s.d.)
0 0.0034
(0.029)
0.0013
(0.016)
0.0008
(0.008)
0.0014
(0.021)
0.0025
(0.026)
312 Australian Journal of Zoology A. Asem et al.

localities is derived from commercialised SFB populations in the
USA, based on similar haplotypes (Fig. 3). These two groups also
possessed the lowest values for the population differentiation
index (F
ST
) between SFB and HED/SKI (Table 3).
Interestingly, we found a single haplotype (H3) connected to
the main haplotype H2 from GSL, consisting of SFB (12.12%),
DAM (30.03%) and MUL (57.58%) localities. All sequences of
DAM and 73.06% of MUL individuals belonged to this
haplogroup, while it held only 10.82% of total SFB sequences.
Generally, the observed haplotype pattern of DAM could suggest
an origin in the SFB, while MUL possessed an intermediate
structure between GSL and SFB (Fig. 3, Tables S2, S3,
Supplementary Material). In contrast, the high values of F
ST
among GSL/SFB and DAM/MUL, and in particular between
SFB and DAM (69.7%) are considerable and cannot corroborate
the suggested network distribution (Table 3). Ordinarily,
introduction from the multiple sources (both GSL and SFB)
could explain this observation. As an alternative hypothesis, the
origin of MUL may be secondary introduction from other
sources, in particular eastern Asia, where A. franciscana cysts
from the Mekong Delta (Vietnam) and Bohai Bay (China) are
easily obtainable in aquaculture markets (Van Stappen et al.
2007; Muñoz et al.2014;Leet al.2018).
The F
ST
value was strongly significant between the two
American populations, GSL and SFB (60.9%) (see Table 3). This
result is similar to previous calculations (59.3%) performed by
Muñoz et al.(2014). The pattern of haplotype frequencies
(Fig. 3), as well as the value of F
ST
, strongly suggest that there is a
high degree of genetic separation between the GSL and SFB
populations.
Typically, the invasive populations have lower genetic
variation in their non-native locations compared with the original
population (Golani et al.2007). Reduction of haplotype variation
and low intraspecific genetic differentiation has also been
0
350
300
250
200
150
100
50
0
25
20
15
10
5
0
0123
MUL SKI
HED
GSL SFB
SSD: 0.283, P = 0.000
Hri: 0.29, P = 0954
SSD: 0.017, P = 0.463
Hri: 0.164, P = 0.703
SSD: 0.002, P = 0.625
Hri: 0.192, P = 0.500
SSD: 0.074, P = 0.164
Hri: 0.308, P = 0.253
SSD: 0.009, P = 0.098
Hri: 0.124, P = 0.109
4
160
140
120
100
80
60
40
20
0
900
800
700
600
500
400
300
200
100
0
400
350
300
250
200
150
100
50
0
5678
0
01
Observed Simulated
23 4 0 1 2 3 4 5
12 3
90123
Observed Simulated
Observed Simulated
Observed Simulated Observed Simulated
Fig. 4. Observed mismatch distributions and their curve fit to simulated models of demographic expansion. MUL, Mulgundawa;
SKI, St Kilda; HED, Port Hedland; DAM, Dampier; GSL, Great Salt Lake; SFB, San Francisco Bay.
Phylogeography of invasive Artemia in Australia Australian Journal of Zoology 313

observed for introduced A. franciscana in Vinh Chau (Vietnam)
as compared with its source population from SFB, likely due to
founder effects (Kappas et al. 2009). In contrast, Eimanifar et al.
(2014) showed that the genetic diversity of invasive Asian
A. franciscana is higher than in GSL and native Asian species.
Similar results have been recorded in some invasive
Mediterranean populations (Hontoria et al.2012; Muñoz et al.
2014). Our results indicate that DAM is a static population with
no genetic variation, which may be the result of a founder effect
and population bottleneck during the process of colonisation. In
general, MUL had the highest genetic diversity among the
populations examined in this work. This finding might be due to
multiple introductions by human-mediated dispersal events or
secondary introductions, although, higher genetic diversity can
be the result of adaptive pressure and/or physiological plasticity
of the exotic population in a non-native region (Dlugosch and
Parker 2008; Ruebhart et al.2008; Vikas et al.2012; Muñoz et al.
2014; Eimanifar et al.2014). We propose that environmental
conditions in new habitats could also have exerted selective
pressure during development of the invasive population.
Two locations from Australia (SKI and HED) and GSL
showed a unimodal structure of mismatch distribution, as well as
a low and non-significant value of SSD and Hri, which indicate a
recent demographic expansion. These findings suggest high
adaptive potential and physiological plasticity of exotic
Australian populations in the new habitats. The demographic
history of MUL and SFB presented a complex structure indicated
by multimodal patterns of mismatch distribution due to
demographic equilibrium. A significant value of SSD (P<0.001)
in MUL confirmed these results, but a non-significant value of
SSD in SFB and a non-significant value of Hri in both
populations highlights the existence of demographic expansion.
The results of this study indicate that the native American and
Australian A. franciscana populations have undergone some
degree of demographic expansion, with the exception of the
DAM population.
In conclusion, our results verify the previous observations of
the colonisation of Australia by invasive A. franciscana
populations. Yet these populations harbour higher levels of
genetic variation than the native America population, in contrast
to other previously studied taxa (see Golani et al.2007).
A. franciscana possesses a faster filter-feeding rate and higher
reproductive rate than the native species (Amat et al.2007;
Sanchez et al.2016). It is also immune to the reduced feeding rate
caused by cestode parasites, contrary to the native populations
(Sanchez et al.2016). These characteristics provide a high
adaptive potential for A. franciscana in new non-indigenous
regions, ultimately resulting in replacement of native species.
Although there are many more populations of A. franciscana in
America, the findings confirmed that Great Salt Lake, Utah, and/
or San Francisco Bay, California, should be the most likely
source for all the current invasion populations outside America.
The utilisation of Artemia in aquaculture production without
regard to potential environmental hazards threatens the
biodiversity of Artemia worldwide.
Conflicts of interest
The authors declare no conflicts of interest.
Acknowledgements
This work was financially supported by the German Academic Exchange
Service (Grant No. A/10/97179). The authors thank Mark Coleman and Brian
Timms for collecting Artemia cyst specimens from Australia.
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