INCO Concerted Action
Artemia Biodiversity:
Current global resources and
their sustainable exploitation
ICA4-CT-2001-10020
Consolidated Scientific Report
Period 1 January 2002-31 December 2004
1
OBJECTIVES
Recent research on the brine shrimp Artemia has indicated the need for integration of basic and
applied knowledge. Triggered by current advances mainly on the fields of ecological/evolutionary
genetics and strain characterization, a global Artemia study was initiated aiming at recording
biodiversity and materializing worldwide interest into a conceptual framework. Under this
perspective, a number of objectives were specifically outlined in order to achieve a concerted set of
methodological and theoretical approaches on most aspects of Artemia biology. According to the
Technical Annex, this global concerted Artemia study consisted of a programme of workshops and
study visits, aiming at technical intercalibration, integration of current ecological and evolutionary
concepts, and uniformous methods to assess population dynamics. Guidelines were to be issued on
sustainable exploitation and strain introduction, intended as tools for authorities to protect biodiversity
and to avert overexploitation and extinction of strains. More in detail, the objectives were as follows:
General objectives:
 Worldwide exchange of expertise and technical intercalibration of Artemia strain
characterization; focus on areas critical for Artemia supplies and/or endangered status of
species (Central Asia, China, South America, Mediterranean); definition and harmonization
of current species/strain concepts;
 Promotion of multidisciplinary approach of Artemia characterization (based on
morphology/morphometry, biometry, life span and reproductive characteristics, physiology,
molecular markers…) for the identification of new strains/species;
 Improve cohesion in Artemia study among EU partners;
 Intensification of scientific EU-DC links and DC-DC links;
 Contribution to Artemia biodiversity preservation and sustainable exploitation, especially in
critical areas;
 Elaboration and refinement of current species concepts;
 Use Artemia as model organism, with principles applicable to other animals facing similar
risks.
Specific scientific and technological objectives:
 Issuing of workshop proceedings with scientific, technical and policy guidelines and
recommendations on species/population description, characterization of new populations,
sustainable exploitation and introduction of allochthonous strains;
 Harmonization (integration/intercalibration) of methods to assess population dynamics,
standing crop size and maximal harvests;
 Introduction of molecular systematic assays to the study of phylogenetic relationships and
biogeographical patterns in the genus;
 Training reports, demonstrating upgrading of level of expertise among DC-partners;
uniformity of practices on Artemia species/population description and on exploitation policy
among DC-partners.
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ACTIVITIES
1. WORKSHOPS
1.1. Deliverable 1: Ghent Global Workshop (February 5-7, 2002)
1.2. Deliverable 2: 1st Regional Workshop (Beijing, China, September 23-26, 2002)
1.3. Deliverable 3: 2nd Regional Workshop (Puerto Varas, Chile, November 16-20, 2003)
1.4. Deliverable 4: 3nd Regional Workshop (Urmia, Iran, September 21-25, 2004)
2. TRAINING VISITING SCIENTISTS (Deliverable 5):
Exchange of expertise and technical training on Artemia strain characterization
Several scientific collaborations have been initiated and led to the establishment of other
national/international projects. Visitors were trained in diverse techniques, focusing mainly on
populations of particular interest. Trainees also had the opportunity to come into contact and discuss
with scientists of hosting laboratory about techniques and methodologies. For overview of trainings of
visiting scientists: see Table.
3. CREATION OF GLOBAL ARTEMIA DATABASE
The global Artemia database intends to be a compilation of ecological and geographical data on
Artemia habitats.
4. ORGANISATION OF CYST BANK
 Updating and expansion of cyst banks managed by INCO members
 Integration of member cyst banks into central database; standardization of data accompanying
cyst samples
5. PROJECT WEBSITE
All information, relevant to the Project, is offered online via a website, linked to and hosted by the
Coordinator’s website (see http://allserv.rug.ac.be/aquaculture/rend/inco.htm). This website contains
general project information (objectives; action plans and partnerships); information about meetings
(list of participants, presentations…), reports, several databases (Artemia sites, cyst bank, ISA
publications…), and is to be used as a forum for information exchange from the Coordinator to the
members and between the respective members.
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OVERVIEW OF TRAININGS VISITING SCIENTISTS
VISITING SCIENTIST
COUNTRY
PARTNER
DURATION (MONTHS)
NO
Host institute: partners 1 and 2: ARC-CLO/DVZ
Visits of INCO partner staff on INCO budget
Mrs. Patricia Beristain
Chile
14
3
Mr. Naser Agh
Iran
11
1½+1½
Mrs. Farzaneh Noori
Iran
11
1½
Mr. Hachem Ben Naceur
Tunisia
8
¾
Dr. Wang Xiaomei
China
10
4¾
Mr. Ramin Manaffar
Iran
11
1¼
Visits of scientists belonging to “associated” INCO partners or funded by other projects
Mr. Naser Agh
Iran
11
3
Mrs. Farzaneh Noori
Iran
11
3
Dr. Gudraz Sadeghi
Iran
11
¼
Mrs. Sui Liying
China
10
2+1
Mrs. Yu Haiying
China
10
9½
Mr. Xin Naihong
China
10
¼
Dr. Nguyen Van Hoa
Vietnam
7
3+¼
Dr. Truong Trong Nghia
Vietnam
7
6½ +2½
Mrs. Nguyen Thi Hong Van
Vietnam
7
3
Mr. Jerome Davis
South Africa
9
17
Prof. Gonzalo Gajardo
Chile
14
¼+¼
Prof. Th. Abatzopoulos
Greece
3
¼
Dr. Iskandar Mirabdullayev
Uzbekistan
¼
Host institute: partner 2: AUTH
Visits of INCO partner staff on INCO budget
Mr. Julio Crespo
Chile
14
2¼
Dr. Horst Kaiser
South Africa
9
3
Dr. Marcos Camara
Brazil
13
2¾
Mrs. T. Hong Van
Vietnam
7
2¾
Dr. Peter Marian
India
15
½
Dr. Alba Papeschi
Argentina
6
1
Dr. Graziella Mura
Italy
4
½
Dr. Gudraz Sadeghi
Iran
11
¼
Visits of scientists belonging to “associated” INCO partners or funded by other projects
Dr. N. Van Hoa
Vietnam
7
¾
Mrs. Yu Haying
China
10
1
Dr. Tom MacRae
Canada
½
Mrs. Olga Ruiz Perez
Spain
4
2
Host institute: partner 3: CSIC
Visits of INCO partner staff on INCO budget
Hachem Ben Naceur
Tunisia
8
2
Juliete Goenaga
Argentina
6
2
Peter Marian
India
15
½
Visits of scientists belonging to “associated” INCO partners or funded by other projects
Graziella Mura
Italy
4
¼
Salvatore Moscatello et al.
Italy
4
4
RESULTS ACHIEVED
1. RESEARCH ACTIVITIES
At the Ghent workshop, Consortium members agreed to focus research on the following geographical
areas and thematic issues. Research results, described below, were to a significant extent produced at
the occasion of the trainings of visiting scientists to the resp. host laboratories.
1. Africa: RU, ARC, AUTH, CSIC, INAT
a) Southern Africa (South Africa, Namibia, Magadascar)
A survey of southern African sites was conducted. During this expedition, 33 Artemia sites were
visited, and biological material from 14 previously unsampled sites could be collected. Overall, cysts
were preserved from 17 new sites. The biological material was characterized during two study visits
at the host laboratory in Greece (AUTH). It was shown that southern Africa has both bisexual and
parthenogenetic Artemia species (a bisexual population was identified for the first time from a salt pan
in Namibia), and that there are mixed populations in this region, in which case both reproductive
modes can be found in one pond.
A thorough literature study revealed that Africa has both sexually reproducing and parthenogenetic
populations of Artemia. There are more populations recorded for the Mediterranean region, but this
may be the result of more research and collecting efforts. Overall, we know of 121 records of Artemia
populations on the continent; species identifications and some basic descriptions (mostly
morphometrics) are available for only 33 populations. An estimated 36% of the populations appear to
be parthenogenetic and 64% are sexually reproducing species. At this stage only a tentative species
list can be presented, as much of the work done on African populations needs verification using
molecular techniques. It is estimated that about 37% of the African continent or 11.3 million km2 are
potential Artemia habitat. Water quality has been recorded for relatively few pans, and although there
are few data, there appears to be a trend for most pans being dominated by Na/Mg and sulphate.
Further surveys need to be done to include more pans. A further review of recently available
information on Artemia sites led to the addition of 65 new records of Artemia habitats for the African
continent.
b) North Africa
In this area, biodiversity studies focused on Artemia populations of Egypt and Tunisia. Laboratory
cross-fertility tests were performed for five bisexual Artemia populations located around the eastern
Mediterranean basin in order to evaluate levels of reproductive isolation among these closely
distributed populations. The results support the view that bisexual Mediterranean populations can be
grouped into an eastern and a western subcluster. Cysts of three parthenogenetic Artemia strains from
saltworks in the Nile Delta were evaluated for their potential use in aquaculture (cyst and naupliar
biometrics, cyst hatching characteristics, fatty acid profile, diapause behaviour). Furthermore, a
number of Egyptian Artemia populations were assayed for the effect of salinity on growth, survival,
morphometrics, maturation and reproductive and life span characteristics. Discriminant function
analysis proved to be a useful tool in determining the differential response of closely related Artemia
populations to different environmental conditions. Investigation of three parthenogenetic strains and
one bisexual population from Egypt revealed significant differences between the asexual populations
and the bisexual one, with parthenogens being more euryhaline. Also the presence of A. salina in the
carbonate Wadi El-Natrun Lake was confirmed. Preliminary results of newly found, uncharacterized
populations revealed the mixed nature (A. salina and parthenogenetic strains) of several Artemia
populations in this region as well as the presence of A. franciscana (based, not exclusively, on
molecular markers). Additionally, and in the light of threats to Artemia biodiversity as a consequence
of pollution with heavy metals, research was done to assess the effect of different cadmium
concentrations on the survival of Egyptian Artemia strains.
5
Intensive cyst sampling has been performed in Tunisian sites, and populations from Tunisia were
studied in detail (biometrics of cysts and nauplii; multivariate analysis of adult morphometrics, lipid
and fatty acid analysis, hatching and diapause characteristics, allozyme study, cross-breeding) and
compared with other Mediterranean strains, which confirmed their taxonomic status of Artemia
salina.
2. China: SRI, ARC, CLO-DVZ, AUTH, UU
a) Contamination of Bohai Bay (PR China) samples with A. franciscana
The uncontrolled introduction of allochthonous Artemia species in China has recently been in the
focus of interest. Standard culture experiments with Artemia samples from the Bohai Bay area in
China, harvested as early as 1980 until now, have been performed in order to perform a preliminary
assessment of the gradual contamination of the autochthonous parthenogenetic populations by
introduced A. franciscana. Results show that from ± 1991 onwards, all available samples are at least
predominantly bisexual, with percentages of males fluctuating between 31 and 50 %. Additionally,
molecular data (species authentication making use of HpaII RFLP of the mitochondrial rDNA)
confirm that the Bohai Bay has been invaded from the early ’90s onwards by A. franciscana. Yet it is
also clear that A. sinica individuals are present. Whether this latter occurrence is natural or artificial
remains to be established; even the co-occurrence of 3 species was found in some cyst samples.
b) Artemia biodiversity on the Tibet-Qinghai plateau
The first purpose of the study on Artemia strains from the Qinghai-Tibet plateau was to test its
adaptation to the local environment, i.c. to test the effect of temperature, salinity and chemical
composition of the hatching and culture medium on the hatching performance of cysts and survival of
the adults. Furthermore, a number of biometrical and chemical characteristics that have previously
been proven for the Artemia tibetiana population from Lagkor Co, Tibet, were confirmed for the new
samples, which were proven to be of mixed origin (parthenogenetic-bisexual). These
strains/populations show a similar large cyst diameter, as described for A. tibetiana, and a high
tolerance to low temperatures, combined with a reduced performance (survival and growth) in natural
seawater. This big cyst size and reduced performance in natural seawater and/or at higher
temperatures might be problematic for application in aquaculture. The very high HUFA pattern, on
the other hand (as in A. tibetiana) shows very promising for its use as live feed.
In a further stage, the mixed status of these Artemia populations was assessed through RFLP analysis
of a 1500 bp mitochondrial rDNA fragment and comparison with the data from the cyst authentication
database, which had been developed before. The construction of this database comprised the
following steps. DNA is extracted from a batch of cysts and a mitochondrial rDNA fragment of 1500
bp is amplified by PCR. In a following step this fragment is digested with 8 different restriction
enzymes (all 4 cutters). The resulting electrophoretic patterns are stored in a database that makes use
of the commercial software GELCOMPAR. Presently the database comprises 133 entries,
corresponding with geographically as well as phylogenetically representative samples. Apart from the
possibility to authenticate unknown samples with the developed technique, the database offers also
the opportunity of a preliminary insight into the molecular phylogeography of the genus Artemia
using one single marker, namely mitochondrial rDNA. During the course of the project, it has been
shown that DNA analysis from a single cyst is a very attractive tool to study old material, kept in cyst
banks, that is no longer hatching.
Based on the RFLP patterns, the species composition of the populations from the resp. Qinghai-Tibet
sites was as follows: in two samples all individual cysts displayed the parthenogenetic pattern, in five
samples uniquely A. tibetiana was found, whereas the other samples proved to be a mixture of
parthenogenetic patterns with A. tibetiana and/or A. sinica patterns in various proportions. Among the
parthenogenetic patterns, one particular HpaII haplotype was dominant, while a second haplotype was
rare (equal to or below 3%). The frequency of the four A. tibetiana haplotypes was variable from
6
sample to sample. However, the parthenogenetic pattern(s) also showed up in samples proving
predominantly bisexual, when being cultured in the laboratory. The presumed close genetic
relationship between the local parthenogenetic and bisexual species, and the intra-species
polymorphism, illustrate the need for cautious interpretation of the results and for a multi-faceted and
well-balanced analytical approach when the issue of possible co-existence of eastern Old World
bisexual and parthenogenetic populations is addressed. Overall the data illustrate that the variability in
this area and even in one lake may be considerable, and that it is indispensable to analyze individuals
(cysts or nauplii) and to combine the genetic data with other type of data.
c) Other areas of continental China
Considerable efforts have been made in pre-project times to inventorize and describe Chinese Artemia
populations. This effort was intensified by the project partners and included a further survey of
(mainly inland) Artemia resources, and sampling of local strains, that were studied primarily as a
function of their potential application in aquaculture (biometrics, hatching and diapause
characteristics, nutritional composition….).
3. Iran: UU, ARC, CLO-DVZ, AUTH, CSIC
a) Zoogeography of Artemia in Iran
A survey was carried out to identify the natural biotopes of Artemia in Iran and to collect ecological
data. Morphometric and life span characteristics of these populations were studied at different
salnities. Results of field observations and laboratory tests indicate that all Artemia populations from
Iran, except for the bisexual Artemia urmiana, are parthenogenetic. The biotopes display a wide
variety in ecological characteristics and comprise big inland salt lakes, small seasonal or permanent
lagoons or salinas, and salty rivers. Generally the density of Artemia is very low in all biotopes and all
of them are more or less oligotrophic. Laboratory tests show that survival, growth rate and
reproductivity decrease with increasing salinity.
b) Present ecological situation of Urmia Lake and resource assessment of its Artemia population
Urmia lake is one of the largest natural resources of Artemia and an internationally protected
ecosystem crucial for avifauna migration. As a consequence of hot, dry years around the turn of the
century, water salinity has recently raised from 220 ppt to near-saturation, which has a detrimental
effect on the Artemia population, as ambient salinities are too high for hatching, survival, and
reproduction; also the phytoplankton composition has drastically changed as a consequence of high
salinities. The effects on the avifauna are accordingly disastrous.
The last years, however, with increased precipitation, have seen a gradual return to conditions more
favourable for Artemia, and hence regular resource assessment was resumed through monthly
sampling at 19 sampling sites of the physico-chemical parameters, primary production and Artemia
population. Results indicate that Artemia cysts hatch in Urmia Lake end of March at the far south and
towards the areas where rivers terminate into the lake. Artemia density reaches a maximum by the end
of July/early August, and is gradually decreasing to very low levels towards winter. Due to the high
salinity level of the lake, no significant differences in Artemia production were observed since 2001.
c) Coexistence of bisexual and parthenogenetic Artemia populations in Urmia Lake region
Urmia Lake is of special interest due to the highly controversial information on the endemic A. urmiana
and the (co-)existence/occurrence of parthenogenetic strains. Field observations and competition tests at
different salinities in the range 15-80 g/l confirmed that both bisexual and parthenogenetic Artemia live
in Urmia Lake. The lake is dominated by bisexual Artemia, whereas the asexual population is restricted
to particular areas of the lake. Artemia appearing seasonally in the lagoons at the peripheries of the lake
are exclusively parthenogenetic. Experiments showed that the parthenogenetic population could grow,
mature and reproduce at very low salinities (15-33 ppt), whereas higher salinities (above 50 ppt) are
required for the bisexual Artemia urmiana to attain sexual maturity. Moreover it was found that a very
low percentage of the larvae hatched from the cysts harvested in coastal areas could give rise to
7
parthenogenetic Artemia, but the larvae hatched from the cyst samples collected from interior regions of
the lake gave rise to bisexual Artemia only. Newly hatched nauplii from the cyst samples collected from
the lagoons grew to exclusively parthenogenetic females. Salinity based competition tests were found to
be a suitable and easy method for isolating parthenogenetic Artemia from bisexual species in a mixed
cyst sample.
d) Genetic diversity of Artemia populations from the Urmia Lake region
The RFLP fingerprinting technique, applied on a 1500 bp mitochondrial rDNA fragment extracted
from adults (see above), was used to study the origin and similarity of Artemia populations from the
Urmia Lake region and other parthenogenetic strains from Iran. Although atypical patterns are
showing up, the results indicate that the bisexual and parthenogenetic Artemia individuals from Urmia
Lake and other sources from Iran display identical restriction patterns, corresponding with the typical
restriction pattern of parthenogenetic strains, which illustrates again their close genetic relationship.
This is in line with literature data supporting the idea that the phylogenetic relationship of A. urmiana
with parthenogenetic forms suggests a recent single common ancestral lineage branching to both A.
urmiana and the asexual forms; other authors suggest that parthenogenetic Artemia has evolved from
A. urmiana. In future it will be interesting to use, in addition to mitochondrial markers, nuclear
markers to analyse individuals from Urmia Lake and its neighbourhood. Ongoing research is studying
in detail the genetic diversity both of the present Artemia populations (biomass) and of historical cyst
samples, using different genetic markers. These data, together with those obtained with samples from
the Qinghai-Tibet Plateau (see above), illustrate once again that a multi-marker approach applied on
sexually mature individuals is needed to unravel the phylogenetic relationship between these Artemia
species.
e) Artemia urmiana applications in larviculture
The potential use of A. urmiana cysts in aquaculture has been evaluated using cyst and naupliar
biometrics, cyst shell structure and buoyancy, lipid analysis and life history traits in different
salinities. A study was also designed to find out a suitable enrichment method and levels for A.
urmiana, aiming to improve its nutritional value with respect to fatty acids and vitamine C in view of
their use in sturgeon larviculture. Results of this series of experiments showed that enrichment with
HUFA + Vit. C always improves performance of sturgeon larvae, compared to those receiving
formulated feed and decapsulated cysts.
f) Pond production of Artemia
Due to the reduced exploitation of Artemia from Urmia Lake as a result of its increased salinity, and
because of the need of hatcheries for Artemia cysts, a project was carried out to evaluate the
possibility of culturing A. urmiana in the provinces around Urmia Lake and at the coasts of the
Persian Gulf. The pilot plant experiments in East Azerbaijan resulted in a satisfactory yield of cysts
(16kg/ww/h/m) and a huge yield of biomass (4.6 ton/h/m). In this context of semi-controlled
environment, the reproduction of the A. urmiana population was studied (effect of salinity and of food
level). Both A. urmiana and parthenogenetic Artemia were cultured in ponds at the vicinity of Lake
Urmia. The project resulted in very good yield of cysts (25 kg/dw/h/m bisexual for A. urmiana and 35
kg/dw/h/m for parthenogenetic Artemia).
4. Mediterranean: AUTH, CSIC, LA SAPIENZA, INAT
a) Artemia franciscana as invasive species in the region
Preliminary screening of Artemia populations in the Mediterranean Basin has raised questions over
the possible presence of A. franciscana (the commonest species for aquaculture purposes) in several
localities, as the result of intentional or non-intentional inoculations through saltern management,
aquacultural (hatchery effluents) and pet market activities, and further regional dispersion by local
coastal avifauna. Joined efforts led to the multidisciplinary identification of A. franciscana
populations on the Mediterranean coastline. The updating of cyst bank and database of Artemia cyst
8
samples showed the availability of cyst samples collected in the Western Mediterranean area from the
beginning of the 80´s until today. The morphometric study of adult males and females, obtained from
original cysts and nauplii under standard culture conditions, and the application of multivariate
discriminant analysis provided evidence on the present distribution of autochthonous brine shrimp
populations as well as of foraneous A. franciscana populations. Autochthonous populations are thus
facing the expansion of the exotic A. franciscana populations.
Moreover, a total of 23 individuals (males and females) from El Pilar and El Estanquillo (Spain)
populations were genetically scored. Following DNA extraction, a 535 bp region of the mitochondrial
16S gene was successfully amplified by the PCR technique. Four restriction endonucleases were
subsequently employed in order to digest the PCR product. These enzymes gave specific restriction
profiles for the discrimination of A. franciscana, A. persimilis and A. salina. They can also distinguish
South from North American A. franciscana. The RFLP analyses of both populations produced
patterns typical of South America A. franciscana, which is in accordance with the characterization
based on morphometry. The combination of traditional (e.g. biometrics, morphometry, crossbreeding) and molecular (RFLP analysis) techniques seems to be a useful tool for studying invasion
patterns and species status in the genus Artemia. Co-occurrence of autochthonous parthenogenetic and
American brine shrimp populations was also found in Morocco (Mar Chica) and France (Aigues
Mortes).
With the above information, a wide prospection for Artemia populations was further organized, with
special attention to Portugal and Italy. Information on Artemia distribution in Italy was revised and
updated according to recent data, and a total of 17 findspots were listed: 12 for A. salina, 5 for
parthenogenetic Artemia. More recently, in winter and spring 2004, a new survey was performed in
insular and peninsular Italy in order to check if changements occurred in some of the Artemia sites,
which were not visited anymore after 2002. This prospection allowed reporting the presence of
introduced A. franciscana in Margherita di Savoia salterns (Apulia province), by using morphological,
morphometrical and molecular criteria, while the prospections in Sardinia and Sicily did not show the
presence of that exotic species. In Sardinia it was possible to verify the presence of bisexual
autochthonous A. salina populations in Su Pallosu and Molentargius salt lagoons, free or co-occurring
with parthenogenetic diploid populations. A sample of cysts collected from the practically abandoned
Carloforte salterns on San Pietro Island produced a mixed bisexual A. persimilis plus parthenogenetic
diploid population. In Sicily the general presence of autochthonous A. salina populations was
observed in the two main saltwork complexes still under exploitation (a number of saltworks are in a
state of gradual transformation into agriculture areas).
The prospection in Portugal showed the general presence of A. franciscana populations in the salterns
still operational in the southern region. The only exception was the presence of an autochthonous
parthenogenetic diploid population in the inland saltern of Rio Maior where exotic populations were
not introduced by man or shorebirds. In Portugal the last few decades there has been a general
tendency into abandonment of saltworks or conversion into rice fields or aquaculture facilities.
b) Study of parthenogenetic Mediterranean populations
The possibility of molecular discrimination between diploid and tetraploid parthenogenetic Artemia
strains was also investigated. Employment of PCR-RFLP analysis in the 16S rRNA region confirmed
the presence of different cleavage patterns between di- and tetraploid asexuals. For example, the Odiel
(Spain) parthenogenetic Artemia population is a mixture of diploid and tetraploid individuals (based
on morphometrical data). Twenty individuals were scored for three enzymes. Individuals described as
diploid produced patterns different from those produced by individuals described as tetraploid. This is
a strong indication that the two groups belong to different strains which reinforces the initial
characterisation using morphometry.
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Parthenogenetic strains in the eastern Mediterranean (especially in Greece) seem to be
phylogenetically structured into two distinctive clades, as gauged by PCR-RFLP analysis. Emphasis
was given to the newly described Artemia populations from Mesi and Kessani. These populations are
morphometrically very similar, but divergent from Citros and Megalon Embolon populations.
However, molecularly they are grouped together with Citros population but are substantially
differentiated from Megalon Embolon.
The differential phenotypic responses of two clones from Megalon Embolon (ME1 and ME2) under
various salinity-temperature combinations revealed a possible partitioning effect due to salinity
elevation. Therefore, clonal divergence in life history traits within parthenogens seems to be an
important component of biodiversity, which deserves further consideration.
5. Central Asia: ARC, CLO-DVZ, La Sapienza, UU, Brad Marden, Elena Bojko
a) Study of distribution of Artemia in lakes of Russia, Ukraine and Kazakhstan, with focus on
South-Siberia
In cooperation with Minsk Academy of Sciences, Belarus, and Tyumen State Agricultural Academy,
Siberian Science-Research and Project-Construction Institute of Fishery, Tyumen, Russia, an updated
list of Artemia sites of Ukraine, Kazakhstan and Russia was made, and a survey of Artemia habitats in
south Siberia was made, resulting in the registration of 90 sites, distributed over the Altai region (total
area of Artemia lakes 1280 km2), Kurgan region (123 km2), Omsk region (92 km2) and Novosibirsk
region (92 km2). Details were collected on topography, hydrology and ionic composition,
phytoplankton and Artemia population dynamics, showing that parthenogenetic populations
predominate in these lakes. Until now bisexual populations have only been found in 3 lakes.
Depending on the available field samples, cyst biometrics and adult morphometrics have been
determined, as well as cytogenetics for a number of populations. Detailed Artemia population
dynamics were monitored for two consecutive years in Medvezhye lake and for one year in Nevidim
and Vishnakovskoye lakes.
A number of available samples (from Ukraina, Kazakhstan and Russia) were examined for SEM
morphology and sex ratio. Information gathered on mode of reproduction and species identification
partly agrees with previous data, though a few controversial points emerged from the analyses of sex
ratio and SEM morphology of the available samples, regarding the exact species status of a number of
samples.
Information about INCO activities was disseminated at the occasion of the “NATO Advanced
Research Workshop (ARW) on Artemia biodiversity in the Newly Independent States”, Moscow,
Russia, 17-19 July 2002. This ARW aimed to bring together leading Artemia scientists from NATO
countries and their counterparts from Newly Independent States (NIS, ex-Soviet Union), to exchange
their views over the distribution of Artemia species and strains over this territory, their genetic
relationship, and the carrying capacity of natural habitats and harvestable amounts of Artemia cysts,
and the methodologies to approach these. It intended to integrate current ecological and evolutionary
concepts, and to uniformize methods to assess population dynamics. The general objectives of this
ARW were thus entirely in line with the objectives of the INCO Artemia Biodiversity project, but
specifically applied according to the geographical and socio-political context of the NIS-states and the
biological reality of the Artemia populations occurring in this area.
b) Study of Artemia colonization of the Aral Sea (Uzbekistan)
The Aral Sea has been claimed to be one of the worst human-induced ecological disasters of the
previous century. Efforts to increase the Aral Sea elevation, to reduce the level of salinity, and reverse
the ecological demise of the region have been largely unsuccessful. Prudent commercial exploitation
of an emerging Artemia population could provide income and employment. It is unknown, however,
whether the current hydrobiological and hydrochemical status and primary productivity of the Aral
10
Sea is sufficient to support a stable Artemia population. Detailed ecological information is clearly
needed in order to evaluate the feasibility and potential benefits of commercial exploitation of
Artemia. Future cooperation with local institutes will focus on the description and characterization of
the Aral Sea Artemia population, the development of a population model for Aral Artemia resource,
and will issue resource management and exploitation recommendations.
6. Latin America: ULL, CLO-DVZ, AUTH, La Sapienza, CSIC, UBA, UMA-X, UFRN
In Latin America Artemia biodiversity issues relate to i) the unconfirmed distribution of Artemia
franciscana and A. persimilis, especially in Chile and Argentina, ii) the differentiation between A.
franciscana populations along a north-to-south transect, iii) the possibility of mixed populations and
hybrid zones and iv) the species status of feral Artemia populations in Brazil.
a) Artemia biodiversity in southern South America.
Both species, A. franciscana and A. persimilis, offer good chances to test the premise of the allopatric
mode of speciation, e.g. populations initially separated by a geographic barrier come into contact and
at this stage reinforce reproductive isolation mechanisms originated in geographic separation. Having
this as a framework, lack of reproductive isolation observed in some highly genetically divergent
populations makes sense, if one thinks that Artemia populations have not completed the second step
of the allopatric mode (sympatry).
Characterization of Artemia populations from Chile using molecular markers (16S rRNA) and RFLP
analysis showed the presence of A. franciscana and A. persimilis. The experiments performed with
RFLP analysis of individual adults (1500 bp mtDNA fragment) from 7 Chilean locations allowed
making the following conclusions. Artemia strains in Chili are either A. franciscana or A. persimilis.
The franciscana strains from Chili are considerably different from those in North America. A.
persimilis seems to inhabit salt lakes up 34°25’ south (Pichilemu). The occurrence of persimilis more
or less halfway Chili was already assumed based on results produced by other methodologies. The
RLFP technique has substantiated this assumption. The results also show that the persimilis strain in
Chili differs in RFLP patterns from those occurring in Argentina; some of the samples, such as those
from Chili, display a double restriction patterns with HapII, indicating that several genotypes might be
present in these samples. New genetic markers (RFLP and mitochondrial DNA sequences) have thus
been added to the genetic tools available for analysis, and the existing database on Chilean resources
has been expanded.
This distribution of both species in Chile was confirmed by SEM morphological study of the frontal
knobs. In Northern Chile, the populations analyzed exhibit a “franciscana” pattern, though
interpopulation variation in pattern and ornamentation is evident, also as compared to the populations
from North America. Central Chilean populations also showed a “franciscana pattern”, with
Pichilemu, however, being closer to the “persimilis pattern”, but showing some atypical
characteristics for this species. In southern Chile, Laguna Amarga and Torres del Paine are both
characterized by a similar appearance, close to “persimilis” but with some particular details
distinguishing them from the specimens from Argentina.
Chile has thus resulted in an interesting study case from an evolutionary biology perspective, and the
various results listed above benefited from the chance of having the two New World species (quite
genetically distant) in the country. According to the data produced by the project, A. persimilis would
have diverged from the ancestral group of species in the Mediterranean 12.6 MY ago, while
separation of A. franciscana from A. persimilis occurred about 3.7. MY ago. The calibration of the
molecular clock on which this extrapolation is based, is arguable, but this estimation is in agreement
with results by other authors and using different methods. The separation of both species is not related
with the Andes formation (a significant barrier that explains their posterior differentiation), at least in
the southern end of the continent (Chile & Argentina). The hypothesis that A. persimilis appeared in
11
Argentina by a founder event, and is close to the ancestral group of species that evolved in the
Mediterranean was confirmed with our data. On the other hand, the hypothesis that A. franciscana
originated at a later stage from A. persimilis, and that it is a derivate species, was also confirmed by
the data.
Next to Chile, the project partners also focused on the presence of A. persimilis and A. franciscana
populations in Argentinean hypersaline ecosystems. A survey was made of Argentinian Artemia sites,
and by means of morphology, biometry of cysts and nauplii, cytogenetics, morphometry of adults,
inter- and intra-populational cross-breeding and RFLP molecular analysis, the species status of each
population was determined.
The information available allows supporting the actual presence of A. franciscana in 5 biotopes and of
A. persimilis in at least 21 biotopes in Argentina. There are also indications for the presence of A.
persimilis in Laguna de Salinas in the Peruvian Puna near Arequipa, and in Laguna de los Cisnes in
the Chilean Tierra del Fuego, which exceeds the “classical” distribution pattern of this species in
Argentina and Chile. Eleven inland and coastal Artemia populations from Argentina and Peru were
genetically scored (PCR-RFLP of 16S rRNA). Preliminary results showed that eight of these
exhibited A. persimilis restriction patterns while the rest showed A. franciscana patterns. Unlike
molecular markers (16S rRNA), morphometric analysis gave substantial evidence for the existence of
hybrid zones between the two species.
Furthermore, the ecological requirements and physiological tolerances of Argentinian A. persimilis
and A. franciscana populations were investigated. The results obtained can allow to predict the
response of each population to environmental variations and to understand the present geographic
distributional pattern of both species in South America. The most relevant results were:
 A. persimilis always showed high mortality at low salinities (30 g.l–1).
 In both species a salinity of 150 g.l–1 is a critical upper limit.
 A. franciscana always showed low survival at low temperature (12ºC), but survived better
than A. persimilis at 21º and 28ºC.
 A. franciscana always showed higher fecundity rates than A. persimilis.
 A. persimilis showed higher growth rates than A. franciscana at all temperatures.
 A. persimilis showed a higher maturation rate than A. franciscana at 12ºC. For the other
temperatures the results were similar in both species.
 In both species temperature increase is associated with increased survival and growth rates
and with a decreased pre-reproductive period and maturation rate.
Also cyst biometrics were determined, including chorion thickness, in order to explain the different
cyst buoyancy levels found for Argentinean A. franciscana and A. persimilis populations. Generally
A. persimilis cysts are bigger than A. franciscana cysts, and hatch into bigger nauplii, especially for
those A. persimilis populations present at southern latitudes or at higher altitudes (colder biotopes),
according to thermo-ecological rules. Quantitative and qualitative identification of PUFA levels in
cyst samples was performed, aiming to correlate these profiles to species-specific conditioning;
marine or inland origin of brines, shoreline, sea level or high altitude biotopes; phenotypic influence
of the diet versus a possible genotypic effect, etc.
Finally, also cytogenetics of Argentinian populations were studied: the number of chromocenters and
the heterochromatin content have been measured in a number of Argentinean populations, and the use
of the percentage of heterochromatin is proposed as a reliable parameter instead of the number of
chromocenters. Also morphological differences in male and female structures involved in the
amplexus in pairs of both species were studied, as well as the development of the compound eye of
Artemia reared under different light conditions.
b) Artemia biodiversity in Brazil
12
Regarding Brazil, there was sound evidence for the origin of the inoculated population in the state of
Rio Grande do Norte being A. franciscana San Francisco Bay (SFB) strain. The molecular genetic
diversity of three A. franciscana populations (Macau, Galinhos and Areia Branca/Grossos) from
northeastern Brazil, was analysed by means of RFLP analysis of their mtDNA. First, these feral
populations were screened in order to confirm if they all belonged to the A. franciscana superspecies.
Following, their northern (San Francisco bay, California, USA) or southern (South America) origin
was determined. And finally the resulting composite genotypes were established. The RFLP data
obtained in this preliminary characterization confirmed that the feral populations of A. franciscana
found in the state of Rio Grande do Norte, northeastern Brazil, belong to the A. franciscana
superspecies. In addition, their proposed origin, from San Francisco Bay cysts, was clearly
demonstrated. This second information corroborated a previous report based on allozyme evidence
derived from the Macau population.
Aiming at the reduction of harvesting pressure in Brazilian saltworks, the feasibility of small-scale
Artemia farming as a complimentary measure to augment the availability of cysts and biomass for use
in local aquaculture is under evaluation in a pilot farm located in the municipality of Grossos in the
State of Rio Grande do Norte.
c) Artemia biodiversity in Mexico
An inventory of Artemia populations in Mexico was made. Preliminary analysis indicates that the
Mexican strains show different RFLP patterns in relation to the North American or South American
strains. Some strains show high similarity to Central American strains. Furthermore different aspects
of strain study were addressed in Mexican Artemia populations:
 HUFA contents and chemical (protein) composition of Artemia cysts and later life stages;
 Concentration of heavy metals Fe, Cu, Mn and Zn in Artemia nauplii and its possible
impact on fish larviculture;
 Biometrics and morphometrics of cysts, nauplii and adults, in relation to latitude of
occurrence and to possible hybridisation;
 Cross-breeding studies.
7. South Asia: IART, ARC, AUTH, CSIC, CTU, UU
Extremely limited information exists on the numerous Artemia sites in India. Substantial efforts still
have to be invested for sampling and genetic characterization of these populations. The project has
addressed Artemia biodiversity in this area by the following lines of research.
An ecological study was conducted of the parthenogenetic Artemia population in the Thamaraikulam
and Puthalam coastal area of South India by weekly sampling in the period 2003-2004. This study
implied the monitoring of the physico-chemical parameters of the salt ponds, the meteorological data,
the primary production, density measurements of the Artemia population, and biometrical
characterization of the cysts and nauplii.
A survey conducted in Tuticorin province on the presence of Artemia revealed that in all places
investigated mixed populations are found of autochthonous parthenogenetic Artemia and introduced
A. franciscana, whereas till 1998 only parthenogenetic Artemia was observed, as proven by sex ratio
analyses. In the period 2002-2004 the average cyst diameter in this area has gradually decreased,
suggesting the gradual decrease of the local parthenogenetic strain. Morphometrical analysis
confirmed that, whereas in some sites the autochthonous diploid parthenogenetic populations can still
be found, other sites display a mixture of parthenogenetic strains and A. franciscana, and others show
the exclusive presence of A. franciscana. Heavy metal concentrations (Zn, Cu, Pb, Hg, Cd) were
registered for saltpans in the Tuticorin province; the effect of heavy metals on the life history and
reproductive characteristics of local Artemia was therefore studied.
13
Furthermore, Artemia strains from Sri Lanka (Puthalalam) and India (Tuticorin and Puthalam) were
compared with North American and Chinese Artemia. The Sri Lanka and Tuticorin strains could be
crossed with A. franciscana, which suggests that these sites have at least partially been contaminated
by this allochthonous species. The Sri Lanka population, for which also cyst biometrics and hatching
characteristics were studied, showed the exclusive presence of A. franciscana.
Finally, a survey of the Artemia populations in Thar Desert, Rajasthan, India, was conducted. Sambar
and Didwana are two inland hypersaline playas located in Thar Desert, known for inland salt
production. Sambar (190 km²) is a shallow basin with a maximum and average depth of 3.00 and 0.61
m respectively, fed by four ephemeral streams, several rivulets and surface run-off. The average
rainfall in the region is 500 mm. Didwana playa covers an area of 16.5 km² at full capacity, and has a
maximum depth of 5 m. It is mainly fed by rainwater. The average annual precipitation in the region
is 330 mm.
The occurrence of blooming populations of parthenogenetic brine shrimp was reported for both sites
decades ago, but by 1977-78 the population had disappeared from Sambar, and Didwana was reported
to be the only existing natural inland biotope of the brine shrimp in India since then. A survey in both
playas, conducted in the period 2000-2004, showed that also in Didwana the Artemia population had
disappeared. Soil and zooplankton samples were collected in different seasons during 2000-2004, and
Cyclops, Brachionus and Moina were found to be the dominant zooplankton forms, but no life stages
of Artemia were found. No cysts were retrieved in both wet (submerged) and dry (shore) bottom
samples. Salinity showed large seasonal fluctuations (from brackish to near-saturation). The reasons
for the total disappearance of brine shrimp from both sites in Thar desert are sofar not clear and
require further field research.
A number of workshops and seminars were organized, focusing on Artemia biodiversity in India:
National Seminar on Status of Artemia Research in India
A national seminar on "Status of Artemia research in India" was organized by IART, Manonmaniam
Sundaranar University on March 22- 23, 2002. 79 papers were presented in the form of oral and
posters. The sessions were on
1. Genetics, Biochemistry, Toxicology and Radiology
2. Ecology, Aquaculture and Commercial production of Artemia
3. Artemia as a tool in biomedical research and biotechnological role of Artemia and
Aquaculture and commercial production of Artemia
Workshop on Characterization of Artemia from saltworks
December 7- 13, 2004
A training workshop was organized on the characterization, culture and use of Artemia from salt
works.
8. Diapause: AUTH, CLO-DVZ, IART, La Sapienza, Tom McRae, Jim Clegg
Artemia cysts are characterized by the occurrence of a dominant heat shock protein protein p26.
Within the framework of this INCO project the p26 mRNA could be extracted from Artemia cysts and
amplified by RT-PCR (occasionally the protocol could be applied to a single cyst). Samples from
eight strains have been subjected to this RFLP technique, showing considerable differences between
franciscana, persimilis and parthenogenetic strains. This observation offers the possibility to start a
phylogeographic study based on a non-mitochondrial marker. In addition, as the p26 protein is
involved in stress resistance, it offers the opportunity to study the relationship between gene
composition (HSP26 genotype) and prevailing environmental circumstances based on cyst material.
Presently sequencing of HSP26 fragments is ongoing (by T. MacRae, Department of Biology,
Dalhousie University, Halifax, Canada).
14
9. Phenotypic plasticity: CSIC, CLO-DVZ, AUTH, INAT, RU, UMA-X, Tom McRae
The adaptation of A. franciscana (SFB strain) to the extreme environmental conditions of Vin Chau
(VC) saltworks, Vietnam, has been fully investigated. Results have demonstrated the high degree of
phenotypic plasticity in A. franciscana as well as its potential for rapid adaptation to a novel
environment. In collaboration with T. MacRae, Department of Biology, Dalhousie University,
Halifax, Canada, the cDNA of 5 A. franciscana sources has been sequenced. The data suggest a single
nucleotide difference between ARC 1258 (from San Francisco Bay, SFB) and the Vietnam strains (the
Vietnam strains are collected in a monsoon area; the inoculum originated from the SFB). This
difference could become a genetic marker for these strains, facilitating ecological studies on the
increase in thermotolerance in the Vietnam strains.
An RFLP method was developed to distinguish the two HSP26 alleles, in such a way that the
homozygote and heterozygote genotypes could be scored. It turned out that the allele frequencies in
the SFB and Vinh Chau (VC, Vietnam) populations are not statistically different; so none of the two
alleles can be used as a marker for SFB or VC individuals.
10. Other biodiversity issues
Presence of rare males in diploid parthenogenetic Artemia populations from the Old World
Rare males were found in 26 diploid parthenogenetic populations from the Old World. These males
are morphologically different from those present in bisexual species. This phenomenon is more
frequent in populations from Central Eurasia (Iran, Iraq, Kazakhstan, Ukraina, Mongolia) and less
frequent or scarce, in peripheral Western (Iberian peninsula, Africa) and Eastern (China) populations.
Although rare males seem useless in reproductive strategies for parthenogenetic females, they are not
sterile in fact. They can mate with and fertilize females from the bisexual Artemia urmiana and A.
sinica, resulting in live offspring. The rare male phenomenon is inferred to help explain the origin of
parthenogenetic strains (polyphyletic origin) from hybridization between two of the Asiatic bisexual
species (A. urmiana and A. sinica) or from lineages leading to present-day Asiatic Artemia, or
alternatively through a monophyletic mechanism similar to the one theoretically argumented in
cladocerans, linked to the transition from cyclic to obligate parthenogenesis.
Research in progress is trying to unveil the status of consecutive hybrid generations produced by
mating rare males of parthenogenetic origin with A. urmiana or A. sinica females, in terms of
postmating isolation, production of abortive and viable offspring, survival rates of the viable
offspring, etc.
2. WORKSHOPS
2.1. Ghent Global Workshop (February 5-7, 2002)
Number of participants INCO partners: 17
Number of participans non-INCO partners: 8
Oral presentations:
a) INCO Members
Sorgeloos Patrick (RUG): Welcome and introduction
Van Stappen Gilbert (RUG): Artemia research at the Artemia Reference Center
Bossier Peter (CLO-DVZ): A database for Artemia authentication
Abatzopoulos Theodore (AUTH): Running research projects at AUTH on Artemia and other
organisms
Amat Francisco (CSIC): Artemia biodiversity: current global resources and their sustainable
exploitation
Nguyen Van Hoa (CTU): Artemia pond culture, strain characterisation and its approach in Vietnam
15
Romdhane Mohammed (INAT): Status of the Artemia biodiversity in Tunisian salt areas
Kaiser Horst (RU): Characterisation of southern African Artemia populations, and the use of Artemia
as a vector for probionts and hormones
Xin Naihong (SRI): Artemia resources and their development in China
Agh Naser (UU): Studies on Artemia populations from Iran
Castro Mejia Jorge (UMAX): Artemia research in the Universidad Autonoma MetropolitanaXochimilco, Mexico
Camara Marcos (UFRN): Artemia research at Universidade Federal do Rio Grande do Norte
(UFRN), Natal, Brazil
Gajardo Gonzalo (ULL): Artemia characterization, species and speciation
Maryan Peter (IART): Research on Artemia in the Institute for Artemia Research and Training
b) Guest Speakers
Boyko Elena, Tyumen State Agricultural Academy - The Siberian Science-Research and ProjectConstruction Institute of Fishery Tyumen, Russia: Authentication of Artemia from Russia
Brendonck Luc, Laboratory Aquatic Ecology, Catholic University of Leuven, Belgium: Adaptations
to life in hyper-oligotrophic ephermeral pools
Clegg Jim, Bodega Marine Laboratory, Section of Molecular and Cellular Biology, University of
California, Davis, Bodega Bay, USA: The diversity of Artemia habitats is reflected in the
biochemical repertoire of this remarkable organism
Criel Godelieve, Department of Anatomy, Embryology and Histology, Ghent University, Belgium:
Morphological tools to help distinguishing Artemia species and populations
De Meester Luc, Laboratory Aquatic Ecology, Catholic University of Leuven, Belgium: Dispersal,
habitat size and genetic differentiation among zooplankton populations
MacRae Tom, Department of Biology, Dalhousie University, Halifax, N.S., Canada: Microtubule
proteins and chaperones: molecular technologies applied to study of the brine shrimp, Artemia
franciscana
Marden Brad, Research and Development, Utah Strategic Alliance, Utah, USA: Recovery of the Great
Salt Lake, USA, brine shrimp (Artemia franciscana) population
2.2. 1st Regional Workshop (Beijing, China, September 23-26, 2002)
Number of participants INCO partners: 24
Number of participans non-INCO partners: 17
Oral presentations:
a) INCO Members
Li Shusheng (SRI): Welcome and Introduction of Salt Research Institute
Sorgeloos Patrick (RUG): Introduction to INCO project
Van Stappen Gilbert (RUG): INCO activities at Artemia Reference Center
Bossier Peter (CLO-DVZ): A rDNA-RFLP database for identifying commercial Artemia samples
Abatzopoulos Theodore (AUTH): Artemia characterisation based on mtDNA RFLP analyses
Amat Francisco (CSIC): Artemia biodiversity: current global resources and their sustainable
exploitation
Mura Graziella (La Sapienza): Use of morphological characters as a tool for species separationRosa
Graciela Cohen de Sanchez (UBA): An overview of the team's activities
Van Hong Nguyen Thi (CTU): Artemia culture in Southeast Asia: emphasis to Vietnam
Kaiser Horst (RU): Preliminary RFLP analysis of mtDNA from southern African Artemia
Sui Liying (SRI): Characterization of Artemia from Qinghai-Tibet Plateau Salt lakes
Yu Xiuling (SRI): Characterization of 4 Tibetan Artemia strains and HUFA profile during
enrichment and starvation
Agh Naser (UU): Coexistence of bisexual and parthenogenetic Artemia populations in Lake Urmia
Thalia Castro (UMAX): Progress in Artemia study at UMA-X
16
Camara Marcos (UFRN): Ecological threats to Artemia franciscana Kellogg (Crustacea; Anostraca)
populations in the coastal saltworks of Rio Grande do Norte, northeastern Brazil
Gajardo Gonzalo (ULL): Artemia diversity and evolution in South America: new results and ideas in
the framework of the Artemia Biodiversity project
Maryan Peter (IART): Status of Artemia production in the coastal salt works of India
b) Guest Speakers
Chen Shao-zhong, Hangu Saltworks, Hangu, Tianjin: The application of Artemia and development of
new Artemia product
Jia Qinxian, Open Laboratory of Saline Lake Resources and Environment, The Chinese Academy of
Geological Sciences, Beijing: Improvement of insect ecdysome on the hatching rate of Artemia
eggs
Kong Fanjing, Research & Development Center of Salt lake and Epithermal Deposits, Chinese
Academy of Geological Sciences, Beijing:Genetic diversity of halophilic organism in Chinese
salt lakes and application prospect for plant salt tolerance genetic engineering
Sun Jingxian, Dalian Fisheries University, Dalian: Some ecological parameters of Artemia
parthenogenetica from Gahai Lake and its use in resource exploitation
Zeng Hui, Life Sciences College, Nankai University, Tianjin: The cloning of the Brine Shrimp DNA
fragments correlated to the Bombyx mori doublesex gene
Zhang Fu, Marine Science and Engineering College, Tianjin Science and Technology University,
Tanggu, Tianjin: Preliminary study on the effect of Na/Mg ratio of artificial seawater and natural
seawater on hatching ability of Artemia cysts
2.3. 2nd Regional Workshop (Puerto Varas, Chile, November 16-20, 2003)
Number of participants INCO partners: 17
Number of participans non-INCO partners: 1
Oral presentations:
a) INCO Members
Gajardo Gonzalo (ULL) and Van Stappen Gilbert (RUG): Welcome and opening comments
Van Stappen Gilbert (RUG): Present studies on biodiversity of Artemia populations in Northern Asia
Bossier Peter (CLO-DVZ): Polymorphism in Artemia species as revealed with mitochondrial and
nuclear markers
Abatzopoulos Theodore (AUTH): Preliminary data on intraspecific genetic divergence between
Artemia franciscana, San Francisco Bay and inoculated populations in Vietnam
Baxevanis Athanasios (AUTH): How do mitochondrially identical Artemia clones respond to different
salinities?
Amat Francisco (CSIC): The use of RFLP 16S rDNA analysis for detecting invasion of Artemia
franciscana in Western Mediterranean region
Amat Francisco (CSIC) : Artemia biodiversity: current global resources and their sustainable
exploitation
Cohen Rosa Graciela (UBA): Record of new Artemia populations and contribution to the
morphological and ecological characterization of some Argentinean populations
Romdhane Mohammed S. (INAT): Strain characterisation and climate effect on Artemia from
Tunisia
Kaiser Horst (RU): Artemia research in southern Africa: A report on a sampling expedition through
parts of South Africa and Namibia
Xin Naihong (SRI): Distribution and characterization of Artemia in Bohai Bay
Agh Naser (UU): Diversity of Artemia populations at Lake Urmia
Castro Thalía (UMAX): Morphological and biochemical characterization of Mexican populations of
Artemia franciscana, and data on reproductive isolation and bioencapsulation of medications
Camara Marcos (UFRN): Preliminary characterization of Brazilian Artemia franciscana populations
by means of mtDNA RFLP analysis
17
Beristain Patricia (ULL): Preliminary comparison of DNA sequences of the cytochrome c oxidase
subunit I (COI) in Chilean Artemia populations
Peter Marian (IART): Report of activities partner 15
b) Guest speaker
Guillermo Chong, Departamento de Ciencias Geológicas, Universidad Católica del Norte, Av.
Angamos 0610, Antofagasta, Chile: The saline domain of Northern Chile
2.4. 3nd Regional Workshop (Urmia, Iran, September 21-25, 2004)
Number of participants INCO partners: 24
Number of participans non-INCO partners: 82
Oral presentations:
Methodological Aspects of Artemia Biodiversity Studies
Abatzopoulos Theodore (AUTH): The contribution of DNA markers in Artemia biodiversity and future
perspectives
Amat Francisco (CSIC): Artemia biodiversity: current global resources and their sustainable
exploitation
Mura Graziella (La Sapienza): General remarks on the usefulness and limits of morphological
characters in species separation within the genus
Artemia Biodiversity in Latin America
Castro Barrera Thalia (UMAX): Cross breeding and morphometric studies in seven Artemia
franciscana strains from Mexico
Gajardo Gonzalo (ULL): Evolutionary biology and Artemia biodiversity in Chile: how much do we
know now?
Artemia Biodiversity in Africa
Kaiser Horst (RUG): Artemia biodiversity in Africa - current status of Artemia distribution with a view
towards further work
Romdhane Mohammed S. (INAT): Biological and biochemical characterisation of Artemia from
Tunisian wetlands
Iran: the Artemia Populations of the Lake Urmia Ecosystem
Abatzopoulos Theodore (AUTH): Artemia urmiana Günther: reproductive and lifespan
characteristics, cyst and naupliar biometrics, HUFA profiles, chorion structure and cyst buoyancy
Seddighi R., Atomic Energy organization of Iran, Iran: Effect of He-Ne Laser on hatching
characteristics of Artemia urmiana
Hafezieh M., Iranian Fisheries Research Organization, IFRO, Iran: Effects of environmental changes on
reproduction mode of Artemia urmiana in Lake Urmia
Noori Farzaneh (UU): Introducing a different enrichment material for improving HUFA levels in
Artemia urmiana nauplii and juvenile
Tabiee O., Azad University of Arsanjan, Iran: The effect of different HUFA enrichment emulsions on
the nutritional value of Artemia urmiana
Eimanifar A., Iranian Fisheries Research Organization, IFRO, Iran: A molecular PCR-RFLP and
morphological study on floating cyst stock of Artemia urmiana from Urmia Lake
Artemia Biodiversity in Asia
Nasiri S.K., University of Baghdad, Iraq: Studies about Artemia from Iraq
Marian Peter (IART): Threat to the Indian Artemia biodiversity by aquaculture and chemical industries:
need for Artemia genome conservation in South Asia
Sultana Razia, Food & Marine Resources Research Center, Pakistan: On the Artemia populations from
Pakistan
Xin Naihong (SRI): Strain characterization of Artemia from 9 inland salt lakes of China
Baxevanis Athanasios (AUTH): Artemia tibetiana: truth and errors on its species status
Bossier Peter and Van Stappen Gilbert (RUG): Evaluation of mixed status of Asian Artemia populations
18
through RFLP-analysis
Mirabdullayev Iskandar, Institute of Zoology, Uzbekistan: Biodiversity and distribution of Artemia in
Uzbekistan
Artemia Populations from Solar Saltworks
M.R. Camara (UFRN): Is small-scale Artemia farming a sustainable alternative to brine shrimp
harvesting in Brazilian saltworks?
H. Nguyen Van (CTU): Adaptation of Artemia franciscana in Vinchau
Sustainable Exploitation of Artemia Resources in Central Asia
G. Van Stappen (RUG), H. Negarestan (Iranian Fisheries Research Organization, IFRO, Iran): Artemia
resources in Lake Urmia, past and present
L. Vdovchenko (Siberian Research and Design Institute of Fisheries, Russia): Artemia in lakes of
Western Siberia
E. Boyko (Siberian Research and Design Institute of Fisheries, Russia): Identification of Artemia from
Russia
A. Kurtulus (INVE, Kazakhstan): Exploitation of Artemia resources from Kazakhstan
Resource Assessment Workshop
B. Marden (Parliament Fisheries LCC, USA): Resource assessment methods applied in GSL, Russia
and Central Asian countries
Naser Agh (UU): Laboratory analysis and calculations method for resource assessment of Artemia
3. CREATION OF GLOBAL ARTEMIA DATABASE
The global Artemia database intends to be a compilation of ecological and geographical data on
Artemia habitats. A Microsoft Access template has been proposed and accepted by the consortium (see
annex I to Consolidated Scientific Report for example of Database file). For each known Artemia site,
the template provides the following data:
 General information of site: Continent, country, state/province, coordinates
 Strain information:
o Species
o Reproduction
o Ploidy level
o Biometrical Data
o Available cyst samples (in resp. cyst banks of Consortium members)
 Artemia site information: type, size, altitude, depth, temperature regime, ionic
composition, salinity, pH
 Literature references
The Artemia site database has been uploaded with information from the following countries (presently
324 sites introduced): Argentina, China, Chile, Cyprus, Greece, Iran, Italy, Mexico, Namibia, South
Africa, Spain, Tunisia.
The database can be downloaded as a zipped Access file from the project’s website at
http://allserv.ugent.be/aquaculture/rend/INCO/sites%20database.htm.
4. ORGANISATION OF CYST BANK
Objectives:
 Updating and expansion of cyst banks managed by INCO members
 Integration of member cyst banks into central database; standardization of data accompanying
cyst samples
The consortium has conceived the organisation and integration of the cyst banks, maintained by the
respective INCO partners, as one of its key tasks. This included the following steps:
19

updating of the respective cyst banks: correction of mistakes in database, elimination of
questionable information etc..
 uniformisation of partner cyst banks according to template ARC cyst bank (see annex II to
Consolidated Scientific Report for example of Cyst bank file)
 integration of partner cyst banks into one Microsoft Access database
This integrated cyst bank can be downloaded from the INCO website as a zipped Access file
(http://allserv.ugent.be/aquaculture/rend/INCO/database.htm). At regular time intervals (e.g. 3monthly), the updated cyst bank is made available through the website. Presently the cyst bank
contains 1615 different entries (= cyst samples), composed of the following partner cyst banks:
 RUG: 1221
 CSIC (Mediterranean group): 143
 AUTH (Mediterranean group): 33
 La Sapienza (Mediterranean group): 22
 UBA: 29
 INAT (Mediterranean group): 10
 UU: 52
 SRI: 71
 UMAX: 34
For each samples, information is provided on:
 geographical origin (country, state, locality, coordinates…)
 strain: species; bisexual or parthenogenetic
 sample: date of arrival, provider, quantity, storage conditions…
 specifications:
o summary of results of tests run on sample (biometrics, hatching, HUFA profile…)
o cyst orders: details on subsamples sent to contact persons for research (identity of
addressee; date of sending; quantity sent…)
5. PROJECT WEBSITE
All information, relevant to the Project, is offered online via a website, linked to and hosted by the
Coordinator’s website (see http://allserv.rug.ac.be/aquaculture/rend/inco.htm); see annex III to
Consolidated Scientific Report for homepage). In the initial phase of the project, access was limited to
project members. Presently access is open, except for those items where information provided is still
provisional. The website is used as a forum for information exchange from the Coordinator to the
members and between the respective members, and offers the following information:
 general project information: project number, scientific EU INCO-DEV framework…
 project objectives
 coordinates of Consortium members
 information on Project workshops:
 coordinates of participants
 book of abstracts oral presentations
 workshop programme with Oral Presentations (in pdf-format)
 action plans and partnerships: overview of task forces within Consortium, working around
specific issues
 progress reports
 ISA Publications: in order to enhance information flow and harmonization of methodology among
INCO members, ‘associated’ INCO members and any interested parties, a list has been made of
all publications, published in the past under the label ‘ISA = International Study on Artemia’. The
‘ISA’ group is an informal working group of laboratories worldwide, involved in Artemia study:
some of the ISA members are partner of the present INCO consortium (partner 1: RUG; 3: AUTH
and 14: ULL); the ISA group further consists of some of the ‘associated’ INCO partners and a
number of other laboratories belonging to European universities. Since 1978 seventy papers have
20




been published under the ISA label, i.e. co-authored by at least two different ISA members. This
list of ISA publications has been added to the INCO website, where some of the publications can
be downloaded full text.
Cysts database: integrated cyst bank database of project partners: see 4
Artemia sites database: see 3 (access limited to project partners)
“Artemia Manual”: “Artemia biodiversity: protocols and guidelines for study and sustainability
(access limited to project partners): see 6”
Publications related to Artemia biodiversity, (co-)authored by consortium members: provisional
list of publications, generated in project’s lifetime, with links to websites of publishers and/or
journals, where abstract and/or full-text version of paper may be retrieved, depending on website
policy.
General items of the website have been translated into Spanish.
6. “ARTEMIA MANUAL”: ARTEMIA BIODIVERSITY: PROTOCOLS AND GUIDELINES FOR
STUDY AND SUSTAINABILITY
In order to promote the visibility of the INCO Artemia Biodiversity Consortium as a group, and to
strengthen the links between Consortium Members beyond the project period, the Consortium
partners agreed to the realization of a joint publication. The Consortium considers this to represent a
direct and essential contribution to the realization of the Project Objectives.
Table of Contents:
Preface
1. Artemia sampling, sample preservation, diapause termination techniques
2. Genetics and molecular markers
1. Cytogenetic analyses and methodologies
2. Cross breeding
3. Genetic approaches to the analysis of Artemia biodiversity
3. Biometrics and morphometrics
1. Cysts and naupliar biometrics
2. Culturing conditions
3. Discriminant analysis on adult Artemia morphology
4. SEM techniques
5. Statistical analysis
4. Phenotypic response under different environmental conditions
5. Sustainable exploitation:
6. Artemia conservation and invasion patterns
Draft versions of the respective chapters can be downloaded from the Project website
(http://allserv.ugent.be/aquaculture/rend/INCO/manual.htm), and are added as annex IV to this
Consolidated Scientific Report.
21
PROBLEMS ENCOUNTERED
(see also Management report)
No major problems were encountered in the scientific realization of this project.
Although invitees for training often originated from disciplines different to those of molecular
genetics and taxonomy, the problems anticipated during their adaptation were either minor or nonexistent. Due to the rather limited period of stay for most invitees, however (generally 3 months or
less), often the invitee left the hosting institute at the moment when he/she was competent to produce
scientific results in an optimal and efficient way.
Moreover, the efficiency of the training stay was highly dependent on the availability of non-INCO
funds, to be used for consumables, which make up a substantial cost in the use of molecular
techniques. It is therefore obvious that the production of scientific results, described above, has been
accomplished using other, non-INCO funds of the hosting laboratories. The same comment is valid
for expenses needed for field sampling, and for other intercalibration and multidisciplinary research
that has been performed within the framework of this INCO project and whenever a major financial
input was required to cover costs for consumables (e.g. SEM observations).
Generally, the lack of budget for explicit South-South exchange (except for participation to
workshops) and training was experienced as a potential hindrance for non-EU partners to maximally
benefit from the realizations of the project.
TECHNOLOGY IMPLEMENTATION PLAN
Not applicable
22
PUBLICATIONS AND PAPERS
Publications in peer-reviewed journals
ABATZOPOULOS, T.J., EL-BERMAWI, N., VASDEKIS, C., BAXEVANIS, A.D., SORGELOOS, P. 2003.
Effects of salinity and temperature on the reproductive and life span characteristics of clonal Artemia
(International Study on Artemia LXVI). Hydrobiologia 492: 191-199.
ABATZOPOULOS, T.J.; KAPPAS, I.; BOSSIER, P.; SORGELOOS, P.; BEARDMORE, J.A. 2002.Genetic
characterization of Artemia tibetiana (Crustacea: Anostraca). Biological Journal of the Linnean Society, 75:
333-344
ABATZOPOULOS, T.J., TRIANTAPHYLLIDIS, G.V., ROEDAKI, N., BAXEVANIS, A.D.,
TRIANTAFYLLIDIS, A., SORGELOOS, P. 2003. Elevated salinities may enhance the recovery of hydrated
heat-shocked Artemia franciscana cysts (International Study on Artemia. LXV). Belgian Journal of Zoology
133 (2): 103-109.
AMAT, F., R.G. COHEN, F. HONTORIA, J.C. NAVARRO. 2004. Further evidence and characterization of
Artemia franciscana (Kellogg, 1906) populations in Argentina. Journal of Biogeography. 31: 1735-1749
AMAT, F., F. HONTORIA, O. RUIZ, A. GREEN, M. SÁNCHEZ, J. FIGUEROLA, F. HORTAS. 2005. The
American brine shrimp as an exotic invasive species in the Western Mediterranean. Biological Invasions.7: 3747.
BAERT, P.; NGUEYN THI NGOC ANH; BURCH, A.; SORGELOOS, P. 2002. The use of Artemia biomass
sampling to predict cyst yields in culture ponds. Hydrobiologia, 477: 149-153
BAXEVANIS, A.D., T.J. ABATZOPOULOS. 2004. The phenotypic response of ME2 (M. Embolon, Greece)
Artemia clone to salinity and temperature. Journal of Biological Research 1: 107-114
BAXEVANIS, A.D., Ν. EL-BERMAWI, T.J. ABATZOPOULOS, P. SORGELOOS. 2004. Salinity effects on
maturation, reproductive and life span characteristics of four Egyptian Artemia populations (International Study
on Artemia. LXVIII). Hydrobiologia 513: 87-100
BOSSIER, P., W. XIAOMEI, F. CATANIA, S. DOOM, G. VAN STAPPEN, E. NAESSENS, P.
SORGELOOS. 2004. An RFLP database for authentication of commercial cyst samples of the brine shrimp
Artemia spp. (International Study on Artemia LXX). Aquaculture, 231: 93-112
CAMARGO, W.N.; BOSSIER, P.; SORGELOOS, P. ; SUN, Y. 2002. Preliminary genetic data on some
Caribbean Artemia franciscana strains based on RAPD’s. Hydrobiologia, 468:245-249
CAMARGO, W.N., J.S. ELY, G. DURAN-COBO, P. SORGELOOS. 2004. Influence of some physicochemical
parameters on Artemia biomass and cyst production in some thalassohaline aquatic environments in the
Colombian Carribean. Journal of the World Aquaculture Society, 35(2): 274-283
CAMARGO, W.N., ELY, J.S., SORGELOOS, P. 2003. Morphometric characterization of thalassohaline
Artemia franciscana populations from the Colombian Caribbean. Journal of Biogeography 30: 697-702.
CASTRO, M.J., T.CASTRO, B, J.L. ARREDONDO, G. CASTRO, M., R. DE LARA, A. Y A. MALPICA, S.
2004. Cross-breeding studies in seven Artemia franciscana strains from México. Journal of Biological Research,
2: 35-41
CASTRO, B.T., CASTRO M.J., CASTRO, J. 2003. Microelements in nauplii of Artemia franciscana (Kellogg,
1906) from six Mexican populations and their relationship with fishes and crustaceans. Hidrobilogica 13(4):
317-320.
CASTRO, B.T., DE LARA, A. R., CASTRO, M.G., CASTRO, M.J., MALPICA, S.A. 2003. Life food in
aquaculture. Contactos 48: 27-33.
23
EL-BERMAWI N., A.D. BAXEVANIS, T.J. ABATZOPOULOS, G. VAN STAPPEN, P. SORGELOOS. 2004.
Salinity effects on survival, growth and morphometry of four Egyptian Artemia populations. (International
Study on Artemia. LXVII). Hydrobiologia 523: 175-188.
GAJARDO, G., J. CRESPO, A. TRIANTAFYLLIDIS, A. TZIKA, A.D. BAXEVANIS, I. KAPPAS, T.J.
ABATZOPOULOS-2004. Species identification of Chilean Artemia populations based on mitochondrial DNA
RFLP analysis. Journal of Biogeography 31(4): 547-555.
JOHN, J.A.C., T.J. ABATZOPOULOS, P.M.MARIAN-2004. Characterization of a new parthenogenetic
Artemia population from Thamaraikulam, India. Journal of Biological Research, 2: 63-74.
KAPPAS, I, T.J. ABATZOPOULOS, N.V. HOA, P. SORGELOOS, J.A. BEARDMORE. 2004. Genetic and
reproductive differentiation of Artemia franciscana in a new environment. Marine Biology 146: 103-117.
MALPICA, S.A., CASTRO B.T., SANDOVAL T.H., CASTRO M.J., DE LARA A. R., CASTRO M.G. 2003.
Composition of the content of fatty acids in three Mexican populations of Artemia franciscana (Kellogg, 1906),
from inland waters, Rev. Biología Tropical 51.
MOSCATELLO, S.; BELMONTE, G.; MURA, G. 2002. The co-occurrence of Artemia parthenogenetica and
Branchinella spinosa (Branchiopoda: Anostraca) in a salt water pond of southeastern Italy. Hydrobiologia,
486:201-206.
MURA, G., B. BRECCIAROLI. 2004. Use of morphological characters for species separation within the genus
Artemia (Crustacea, Branchiopoda). Hydrobiologia, 520: 179-188.
PASTORINO, X.I., MARSCHOFF, E., COHEN, R.G. 2002. Reproductive and brood cycles of Artemia
persimilis Piccinelli & Prosdocimi from Colorada Chica Lake (Province of La Pampa, República Argentina),
under laboratory conditions. Hydrobiologia 486: 279-288.
SARABIA, R., DEL RAMO, J., VARÓ, I., DIAZ-MAYANS, J., TORREBLANCA, A. 2002. Comparing the
acute response to cadmium toxicity of nauplii from different populations of Artemia. Environ. Toxicol. Chem.,
21(2): 437-444.
VAN STAPPEN, G., SUI, L., XIN, N., SORGELOOS, P. 2003. Characterisation of high-altitude Artemia
populations from the Qinghai-Tibet Plateau, PR China. Hydrobiologia, 500: 179-192.
VARÓ, I., NAVARRO, J.C., AMAT, F.; GUILHERMINO, L. 2002. Characterization of cholinesterases and
evaluation of the inhibitory potential of chlorpyrifos and dichlorvos in Artemia salina and Artemia
parthenogenetica. Chemosphere 48 : 563-569
VARÓ, I., SERRANO, R., PITARCH, E., AMAT, F.; LÓPEZ, F.J., NAVARRO, J.C. 2002. Bioaccumulation
of chlorpyrifos through an experimental food chain. Study of protein HSP70 as biomarker of sublethal stress in
fish. Arch. Environ. Contam. Toxicol., 42: 229-235.
Publications in other journals and magazines
CAMARA, M. R. 2004. Biomassa de Artemia na carcinicultura: repercussões ambientais, econômicas e sociais.
Panorama da Aqüicultura 82(14): 40-45.
CAMARA, M. R. 2004. Cistos de Artemia: oscilações globais de produção, mistérios científicos e desafios
tecnológicos. Panorama da Aqüicultura 63(14): 24-29.
CAMARA, M. R., P. A. BENTO CARNEIRO MONTEIRO, L. GARCIA REIS, M. A. FREIRE DA COSTA
JÚNIOR. 2004. Farming Artemia in a multi-cycle culture system in northeastern Brazil. World Aquaculture
2(35): 40-42.
24
Chapters in books
DHONT, J.; SORGELOOS, P. 2002. Applications of Artemia: 251-277. In: Artemia: Basic and Applied
Biology, Abatzopoulos T. J. et al. (eds), Kluwer Academic Publishers, Dordrecht, The Netherlands
DHONT, J., VAN STAPPEN, G. 2003. Biology, Tank Production and Nutritional Value of Artemia. 65-121. In:
Live feeds in marine aquaculture. J.G. Støttrup, L.A. McEvoy (Eds). Blackwell Science Ltd. 318 pp.
GAJARDO, J.; ABATZOPOULOS, T.J.; KAPPAS, I.; BEARDMORE, J.A. 2002. Evolution and Speciation.
225-250. In: Artemia: Basic and Applied Biology, Abatzopoulos T. J. et al. (eds), Kluwer Academic
Publishers, Dordrecht, The Netherlands
VAN STAPPEN, G. 2002. Zoogeography: 171-224. In: Artemia: Basic and Applied Biology. Abatzopoulos, T.
J. et al. (eds). Kluwer Academic Publishers, Dordrecht, The Netherlands
VAN STAPPEN, G. 2003. Production, Harvest and Processing of Artemia from Natural Lakes. 122-144. In:
Live feeds in marine aquaculture. J.G. Støttrup, L.A. McEvoy (Eds). Blackwell Science Ltd. 318 pp.
Theses
ATHANASIADIS, G. 2003. The use of discriminant analysis based on morphometry: discriminating
parthenogenetic Artemia populations in N. Aegean coastal saltworks. BSc. Thesis, Aristotle University of
Thessaloniki, Department of Genetics, Development and Molecular Biology.
BEN NACEUR HACHEM. 2004. Contribution a la caractérisation biologique et biochimique de trois
populations tunisiennes d’artemia. Mémoire de mastère production et Ecosystèmes Aquatiques INAT 87p.
BEN TAARIT MOUNA. 2004. Valorisation de la plante de bourrache (Burago officinalis) en tant qu’adjuvant
aquacole d’enrichissement d’artemia. Mémoire de DEA FST, 98p.
BOURIGUA NAWZET. 2004. Contribution à l’étude chimique et biochimique et a la caracterisation
spatiotemporelle de la microalgue Dunaliella salina de la Tunisie. Mémoire de mastère production et
Ecosystèmes Aquatiques INAT. 111p.
CAMARGO, W. 2002. Characterization of Artemia populations from Colombia for use in aquaculture. PhD
Thesis in Applied Biological Sciences, Ghent University, Belgium.
CASTRO, M.J. 2004. Morphometric and reproductive (hybridisation) characterisation of seven Artemia
franciscana populations in Mexico, in relation to its habitat and latitude. (in Spanish; abstract in Spanish and in
English). Master Thesis, Universidad Autónoma Metropolitana - Unidad Iztapalapa, Mexico D.F., Mexico, 116
pp.
CASTRO ARAUJO, A. 2003. Taxonomic characterization of three populations (Real de Salinas, Campeche,
Yavaros, Sonora, and Cuatro Ciénegas, Coahuila) of Artemia franciscana in Mexico. BSc. Thesis, Universidad
Autónoma Metropolitana – Unidad Xochimilco, Departamento El Hombre y su Ambiente, Mexico
CHLAIFA HOUDA. 2003. Contribution à l’étude de la composition biochimique et de l’effet bactéricide d’une
souche de la microalgue Dunaliella salina issue de sebkha Eladhibet, Mémoire de mastère production et
Ecosystèmes Aquatiques INAT 80p.
CRESPO SOTO, J.E. 2003. Genetic characterization of Chilean Artemia populations (Crustacea, Anostraca) :
mitochondrial DNA assessment by PCR-RFLP. MSc Zoology Thesis. University of Concepción, Department of
Zoology, Chile.
DELIOPOULOS, G. 2004. Artemia: differential response of two mitochondrially similar clones at various
temperature-salinity combinations, Diploma Thesis, AUTH.
EL-BERMAWI, N. 2003. Determination and identification of biological characteristics of Artemia populations
from the Egyptian Nile delta for application in aquaculture. PhD Thesis. Ghent University, Faculty of
Agricultural and Applied Biological Sciences, 177 pp.
25
GHLALA ADNANE. 2003. Différenciation génétique et morphometriques de quelques populations tunisiennes
d’artemia. Mémoire de DEA genétique et Ressources Animale, INAT 90p.
HAMROUNI SOUHIR. 2004. Caractérisation biochimique et electrophoretique de quatre populations
d’Artemia de Tunisie, Mémoire de mastère production et Ecosystèmes Aquatiques INAT, 85p.
MALAR, D. 2004. Studies on the natural and inoculated Artemia franciscana population in different salt pans
of K.K. District. Ph.D Thesis, M.S. University, IART, 199 pp.
MALPICA SANCHEZ, A. 2002. Estructura y Composición Química de los Quistes de Artemia franciscana de
Diferentes Poblaciones Mexicanas. MSc Thesis. Facultad de Ciencias, Universidad Autónoma MetropolitanaXochimilco, México. 52 pp.
MARKATZINOU, E. 2003. Detection of invasive Artemia species in Mediterranean Basin by using
discriminant analysis. BSc. Thesis, Aristotle University of Thessaloniki, Department of Genetics, Development
and Molecular Biology.
NGUYEN VAN HOA, 2002. Seasonal farming of the brine shrimp Artemia franciscana in artisanal salt ponds
in Vietnam: effects of temperature and salinity. PhD Thesis in Applied Biological Sciences, Ghent University,
Belgium, 184 pp.
PASTORINO, X.I. 2003. Caracterización morfológica y reproductiva de Artemia persimilis (Crustacea
Branchiopoda: Anostraca) de la laguna Colorada Chica (provincia de La Pampa). PhD Thesis, Universidad de
Buenos Aires, Departamento de Biodiversidad y Biologia Experimental
PUNITHA M.J. 2003. Studies on the use of marine algae and ayurvedic products as feed for the brine
shrimp Artemia franciscana. Ph.D Thesis submitted to M. S. University (2003) pp. 182.
SOTO MURRIETA, O. 2003. Content of magnesium, calcium, sodium, potassium and phosphorus in Artemia
franciscana, adult stage, of five Mexican populations and their importance in aquaculture. BSc. Thesis,
Universidad Autónoma Metropolitana – Unidad Xochimilco, Departamento El Hombre y su Ambiente, Mexico
TZIKA, A. 2003. Genetic identification of Artemia species based on mtDNA-RFLP analysis. BSc. Thesis,
Aristotle University of Thessaloniki, Department of Genetics, Development and Molecular Biology.
VIMALA LESLY, J. 2004. Studies on the effects of heavy metals on Artemia spp from Tuticorin. Ph.D Thesis,
M. S. University, IART, 214 pp
WORKU JIMMA HORRA, 2002. Characterisation study of parthenogenetic and bisexual Artemia strains of the
Lake Urmia area, Iran, in function of their possible niche partitioning. MSc Thesis in Aquaculture, Ghent
University, Belgium, 64 pp.
ZARAGOZA RÍOS, J. 2003. Taxonomic characterization of three populations (Juchitan, Oaxaca, Las Salinas,
San Luis Potosí and Texcoco, State of Mexico) of Artemia franciscana in Mexico. BSc. Thesis, Universidad
Autónoma Metropolitana – Unidad Xochimilco, Departamento El Hombre y su Ambiente, Mexico
Presentations in conferences
For presentations at the various INCO workshops, see above “RESULTS ACHIEVED; 2. WORKSHOPS”
Presentations at other conferences and meetings:
ABATZOPOULOS, T.J., BAXEVANIS, T. 2002. The use of multidisciplinary approaches for characterizing
and biomonitoring Artemia populations: uniformity of practices. Oral communication at NATO Advanced
Research Workshop “Artemia Biodiversity in the Newly Independent States: Current Global Resources and
their Sustainable Exploitation”, 7-19 July 2002, Moscow, Russia
ABATZOPOULOS, T.J., TRIANTAPHYLLIDIS, G.V., ROEDAKI, N., BAXEVANIS, A.D.,
TRIANTAFYLLIDIS, A., SORGELOOS, P. 2002. Elevated salinity enhances the thermotolerance of hydrated
26
Artemia cysts. (International Study on Artemia. LXV). 9th International Congress on the Zοοgeography and
Ecology of Greece and Adjacent Regions (ICZEGAR), Thessaloniki, May 2002
AMAT, F., HONTORIA, F., RUIZ, O., SÁNCHEZ, M., GREEN, A., FIGUEROLA, J., HORTAS, F. 2003.
Artemia franciscana como especie exótica invasora en el Mediterráneo Occidental. . I Congreso Nacional sobre
Especies Exóticas Invasoras. “EEI 2003”. Universidad de Leon, España, 4 – 7 junio 2003.
ARBASETTI, C., SOTELANO, P., GOENAGA, J., COHEN, R.G. 2003. Efecto de shocks salinos en distintas
etapas del ciclo ontogenético de dos poblaciones argentinas del crustáceo Artemia (Branchiopoda: Anostraca).
V Jornadas Nacionales de Ciencias del Mar. XIII Coloquio Argentino de Oceanografía,² Mar del Plata,
Argentina, 8-12 December, 2003.
BEN NACEUR H., ROMDHANE M.S , BEN REJEB JENHANI A ., EL CAFSI. 2004. Evaluation de la qualité
et amélioration des performances des cystes de trois souches Tunisiennes d’Artemia.(IRESA-2004)
BEN NACEUR H., ROMDHANE M.S., EL CAFSI. 2004. Fatty acid composition of three popoulation of
artemia from Tunisia.(Libya-2004)
BEN NACEUR H., ROMDHANE M.S., BENE REJEB JENHANI A., EL CAFSI M. 2004. Etude
morphométrie de quatre populations tunisiennes d’Artémia et test de croisement entre différentes espèces
tunisienne, espagnole et marocaine. 7ème Congrès Tunisien des Sciences de la Mer, 18 au 23 décembre 2004,
Zarzis (Tunisie)
BEN TAÂRIT M., EL CAFSI M., ROMDHANE M.S. 2004. Valorisation des cystes de l’Artemie (Artemia
Salina) en provenance d’un site tunisien : La Sebkha Eladhibet. IRESA-2004.
BOURIGA N., ROMDHANE M.S., EL CAFSI M. 2004. L’effet de salinité et de l’intensité lumineuse sur la
composition lipidique de Dunaliella salina de sebkha Lehdhibett. 7ème Congrès Tunisien des Sciences de la
Mer, 18 au 23 décembre 2004, Zarzis (Tunisie)
BERISTAIN, P., S.M. FUNK, G. GAJARDO. 2004. Divergencia genética entre poblaciones chilenas del
crustaceo Artemia estimada a partir de secuencias del gen mitocondrial CO-I. XXXVII. Reunión Anual
Sociedad Genética de Chile, 27-29 October 2004, Viña del Mar, Chile.
BERISTAIN, P., GAJARDO, G., BOSSIER, P., SORGELOOS, P, SAGREDO, B. 2002. Caracterizazión
preliminar molecular de poblaciones chilenas de Artemia (Preliminary molecular characterisation of Chilean
Artemia strains. Annual meeting of the Chilean Society of Genetics, Antofagasta, Chile, 28-30 October 2002
BOSSIER, P., WANG XIAOMEI, CATANIA, F., DOOMS, F., VAN STAPPEN, G., SORGELOOS, P. 2002.
A rDNA-RFLP database for identifying commercial Artemia samples. Oral communication at NATO Advanced
Research Workshop “Artemia Biodiversity in the Newly Independent States: Current Global Resources and their
Sustainable Exploitation”, 7-19 July 2002, Moscow, Russia
CAMARA, M. R., 2003. Towards a sustainable Artemia industry in northeastern Brazil. In: Book of Abstracts,
Aquaculture 2003, Salvador, Brazil. World Aquaculture Society, Baton Rouge, Louisiana, USA. p. 148.
CAMARA, M. R. 2004. Caracterização da produção de cistos e biomassa de Artemia na região salineira do Rio
Grande do Norte (Brasil). Resumos, VI Simpósio Brasileiro sobre Cultivo de Camarão, Natal, 2004, p. 30,
Associação Brasileira de Criadores de Camarão.
CASTRO, T. 2003. Evaluation of crude protein in three stages of development of Artemia franciscana from
different Mexican populations. XXXIV National Congress of sciences and technology in foods. Pachuca,
Hidalgo, México, September, 2003.
CASTRO, B.T. 2004. Cultivo de Artemia para la acuicultura” Ciclo de Conferencias de la VII Semana de
Hidrobiología. May 2004. UAM-Iztapalapa, DF, México
CASTRO, B.T. , CASTRO, M.J. , CASTRO, M.G. , MALPICA, S.A. , DE LARA, A.R. 2002. Manejo del
Recurso Artemia para la Acuicultura” Primer Congreso Internacional “Ecología y Procesos Productivos”.
Universidad Autónoma Metropolitana-Xochimilco, México, D.F., 10-11 October 2002.
27
CASTRO, T.; CASTRO, J, MIRAMONTES, B., CASTRO, J. 2002. Concentración de Fe, Cu, Mn y Zn en
nauplios de Artemia franciscana de México y su relación con los requerimientos del bagre, trucha y de
camarones peneidos. Aquamar Internacional. 6th Latin American Conference and Exposition, Cancún, Quintana
Roo, México, 3-6 September 2002.
CASTRO, M.G., DE LARA, A.R., CASTRO, M.J., MALPICA, S.A, CASTRO, 2002. B.T. Artemia, un
organismo planctónico utilizado como agente bioencapsulante de pefloxacino para el tratamiento de infecciones
por Pseudomona aeruginosa”. 12ª Reunión Nacional de la Sociedad Mexicana de Planctonología y 5ª
Internacional Meeting of the Mexican Society of Planktology; Jalapa, Veracruz, México. 6-9 May, 2002.
CASTRO, T., MALPICA, A., SANDOVAL, H., CASTRO, J., CASTRO, G., DE LARA, R. 2002. Composición
del contenido de ácidos grasos en tres poblaciones mexicanas de Artemia franciscana (Kellog, 1906) de aguas
epicontinentales. Aquamar Internacional. 6th Latin American Conference and Exposition, Cancún, Quintana
Roo, México, 3-6 September 2002.
CAVALCANTE, P. H. DE OLIVEIRA, I. S. DE SOUZA, M. F. COSTA, F. R. S. DE SOUZA, T. DE M.
CABRAL, M. R. CAMARA. 2004. Utilização da macroalga Gracilaria cervicornis e da cianobactéria Spirulina
maxima como dietas complementares no cultivo de Artemia franciscana Kellogg (Crustacea; Anostraca) em
escala laboratorial. Resumos, VI Simpósio Brasileiro sobre Cultivo de Camarão, Natal, 2004, p. 31, Associação
Brasileira de Criadores de Camarão.
CAVALCANTE, P. H. DE O., I. S. DE SOUZA, M. F. COSTA, F. R. S. DE SOUZA, T. DE M. CABRAL, M.
R. CAMARA. 2004. Reprodução de Artemia franciscana Kellogg (Crustacea) em cultivo à base de Gracilaria
cervicornis e Spirulina. In: Resumos, XXV Congresso Brasileiro de Zoologia, Brasília, 2004, pp. 55-56,
Sociedade Brasileira de Zoologia.
COSTA, M. F., F. R. S. DE SOUZA, M. R. CAMARA. 2004. Utilização da macroalga Gracilaria cervicornis e
da cianobactéria Spirulina maxima no cultivo de Artemia franciscana (Crustacea) em escala laboratorial.
Resumos, XV Congresso de Iniciação Científica da UFRN, Natal, 2004, Universidade Federal do Rio Grande do
Norte.
COSTA JÚNIOR, M. A. F., CAMARA, M.R., REIS, L.G., OLIVEIRA, C.P.R. 2002. Maricultura e
extrativismo de Artemia (Crustacea; Anostraca) na região de Grossos, Rio Grande do Norte: uma análise
comparativa. In: XIII Congresso de Iniciação Científica da UFRN, XIII Congresso de Iniciação Científica da
UFRN, Natal, 2002, vol. 1, Natal: UFRN, Brazil
COSTA, M. F., L. G. REIS, P. A. B. C. MONTEIRO, M. R. CAMARA. 2004. Avaliação da performance
reprodutiva de Artemia franciscana (Crustacea, Anostraca) em sistema de cultivo multifásico. Resumos, VI
Simpósio Brasileiro sobre Cultivo de Camarão, Natal, 2004, p. 33, Associação Brasileira de Criadores de
Camarão.
CRESPO, J.E., GAJARDO, G., ABATZOPOULOS, T., TRIANTAFILLIDIS. 2002. A. Artemia en Chile:
Caracterizazión genética del DNA mitocondrial por PCR-RFLP (Artemia in Chile: Genetic characterisation of
mitochondrial DNA by PCR-RFLP). Annual meeting of the Chilean Society of Genetics, Antofagasta, Chile,
28-30 October 2002
GOENAGA, J., MEDINA, G., AMAT, F., COHEN, R.G., 2003. Efectos combinados de salinidad y temperatura
sobre la supervivencia y edad de madurez sexual en dos poblaciones argentinas del crustáceo Artemia
(Branchiopoda: Anostraca). V Jornadas Nacionales de Ciencias del Mar. XIII Coloquio Argentino de
Oceanografía , Mar del Plata, Argentina, 8-12 December, 2003
HAMROUNI S., ROMDHANE M.S., EL CAFSI M. 2004. Essais de séchage de cystes d’Artémia salina : effets
sur la qualité et les performances. 7ème Congrès Tunisien des Sciences de la Mer, 18 au 23 décembre 2004,
Zarzis (Tunisie)
LEAL, A. S. R., CAMARA, M.R., DE SIQUEIRA, W.P. 2002. Caracterização reprodutiva das populações de
Artemia franciscana (Anostraca: Artemiidae) na região salineira do RN. I. Galinhos, Guamaré e Macau. In:
XII Congresso de Iniciação Científica da UFRN, XII Congresso de Iniciação Científica da UFRN, Natal, 2002,
vol. 1, Natal: UFRN, Brazil
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LIPKO; P., PAPESCHI, A.G., COHEN, R.G. 2002. Análisis citogenético de tres nuevas poblaciones de A.
franciscana (Crustacea, Branchiopoda: Anostraca) en las provincias argentinas de La Rioja, Córdoba y San
Luis. XXXI Congreso Argentino de Genética, La Plata, 17-20 September, 2002
LIPKO, P.; PAPESCHI, A.G., COHEN, R.G. 2002. Cytogenetic analysis of four argentine populations of
Artemia (Crustacea, Branchiopoda: Anostraca). IV Jornadas de la Sociedad Argentina de Biología, Buenos
Aires, 6 December, 2002
LIPKO, P., A.G. PAPESCHI, R.G.COHEN. 2004. Variación en el contenido de heterocromatina en cinco
poblaciones de Artemia (Crustacea, Branchiopoda, Anostraca). XXXIII Congreso de la Sociedad Argentina de
Genética. 26-29 September, 2004, Malargüe, Mendoza Province, Argentina
MALPICA, S.A., CASTRO, B.T., SANDOVAL, H., CASTRO, M.J., CASTRO, M.G., DE LARA, A.R. 2002.
Composición química de quistes y huevos descapsulados de seis poblaciones mexicanas de Artemia
franciscana”. Aquamar Internacional. 6th Latin American Conference and Exposition, Cancún, Quintana Roo,
México, 3-6 September 2002.
MALPICA, S. A. , VALIENTE, M.E. , CASTRO, B.T. , DE LARA, A.R. , CASTRO, M.J , CASTRO, M.G.
2002. Estudio de la estructura externa de quistes y huevos descapsulados de seis poblaciones mexicanas de
Artemia franciscana, con microscopio electrónico de barrido” 12ª Reunión Nacional de la Sociedad Mexicana
de Planctonología y 5ª Internacional Meeting of the Mexican Society of Planktology, Jalapa, Veracruz, México.
6-9 May, 2002
MARKATZINOU, E., ATHANASIADIS, G., BAXEVANIS, A.D., ABATZOPOULOS, T.J. 2003.
Characterization of two new Artemia parthenogenetic populations from Thrace (Greece): morphometrical
analysis. 11th Panhellenic Congress of Ichthyologists, Preveza, 10-13 April 2003.
MARYAN, P. 2002. National Seminar on "Status of Artemia research in India", organized by IART,
Manonmaniam Sundaranar University, March 22- 23, 2002
MURA, G., F. AMAT, T.J. ABATZOPOULOS, S. MOSCATELLO. 2004. First record of Artemia franciscana
in an Italian saltwork. 5th International Large Branchiopod Symposium, Toodyay, Western Australia, 16-20
August 2004.
OLIVEIRA, C. P. R., CAMARA, M.R., COSTA, M.A.F., REIS, L.G. 2002. Utilização de Artemia franciscana
(Crustacea, Anostraca) em bioensaios de monitoramento de qualidade ambiental aquática. In: XIII Congresso
de Iniciação Científica da UFRN, XIII Congresso de Iniciação Científica da UFRN, Natal, 2002, vol. 1, Natal:
UFRN, Brazil
OLIVEIRA, C. P. R., CAMARA, M.R., REIS, L.G. 2003. Utilização de náuplios de Artemia franciscana
(Crustacea, Anostraca) no biomonitoramento aquático do litoral urbano da cidade de Natal (RN). In: Resumos,
XIV Congresso de Iniciação Científica da UFRN, XIV Congresso de Iniciação Científica da UFRN, Natal,
2003, vol. 1, Natal: UFRN.
REIS, L. G., CAMARA, M.R., OLIVEIRA, C.P.R. 2003. Monitoramento do estado reprodutivo, modo de
reprodução e produtividade de cistos e biomassa de Artemia franciscana (Crustacea, Anostraca) em sistema de
cultivo multifásico. In: Resumos, XIV Congresso de Iniciação Científica da UFRN, XIV Congresso de
Iniciação Científica da UFRN, Natal, 2003, vol. 1, Natal: UFRN.
REIS, L. G., CAMARA, MR.; OLIVEIRA, C.P.R.; COSTA, M.A.F. 2002. Avaliação da performance
reprodutiva de Artemia franciscana (Crustacea, Anostraca) em sistema de cultivo multifásico. In: XIII
Congresso de Iniciação Científica da UFRN, XIII Congresso de Iniciação Científica da UFRN, Natal, 2002, vol.
1, Natal, UFRN, Brazil
ROMDHANE, M.S. 2003. Biochemical characterisation of Artemia. Regional Workshop on Marine Science,
Tripoli, Libya.
SIQUEIRA, W. P. DE,. CAMARA, M.R., LEAL, A.S.R. 2002. Caracterização reprodutiva das populações de
Artemia franciscana (Anostraca: Artemiidae) na região salineira do RN. II. Areia Branca e Grossos. In: XII
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Congresso de Iniciação Científica da UFRN, XII Congresso de Iniciação Científica da UFRN, Natal, 2002, vol.
1, Natal: UFRN, Brazil
SORIANO-SANTOS, J., S.A. MALPICA, C.V. GARCÍA, B.T. CASTRO. 2004. Fraccionamiento de las
proteínas de embrión descapsulado de Artemia franciscana por extracción secuencial con disolventes. III
Congreso Internacional y XIV Congreso nacional de Ingeniería Bioquímica. March 31-April 2, 2004, Veracruz,
México.
SOUZA, I. S. DE, P. H. DE O. CAVALCANTE, M. F. COSTA, F. R. S. DE SOUZA, M. R. CAMARA. 2004.
Utilização da levedura Saccharomyces cerevisiae e da cianobactéria Spirulina maxima como dietas
complementares no cultivo laboratorial de Artemia franciscana Kellogg (Crustacea; Anostraca). Resumos, VI
Simpósio Brasileiro sobre Cultivo de Camarão, Natal, 2004, pp. 51-52, Associação Brasileira de Criadores de
Camarão.
SOUZA, F. R. S. DE, M. F. COSTA, M. R. CAMARA. 2004. Utilização da levedura Saccharomyces cerevisiae
e da cianobactéria Spirulina maxima no cultivo de Artemia franciscana (Crustacea) em escala laboratorial.
Resumos, XV Congresso de Iniciação Científica da UFRN, 2004, Universidade Federal do Rio Grande do
Norte.
SOUZA, I. S. DE, P. H. DE O. CAVALCANTE, M. F. COSTA, F. R. S. DE SOUZA, M. R. CAMARA. 2004.
Utilização da levedura Saccharomyces cerevisiae no cultivo de Artemia franciscana Kellogg (Crustacea;
Anostraca). Resumos, XXV Congresso Brasileiro de Zoologia, Brasília, 2004, p. 55, Sociedade Brasileira de
Zoologia.
VAN STAPPEN, G., SUI LIYING, WANG XIAOMEI, BOSSIER, P., SORGELOOS, P. 2002. Status of
research of Artemia populations from Tibet, PR China. Oral communication at NATO Advanced Research
Workshop “Artemia Biodiversity in the Newly Independent States: Current Global Resources and their
Sustainable Exploitation”, 7-19 July 2002, Moscow, Russia
VAN STAPPEN, G., SUI, L., XIN, N., SORGELOOS, P. 2003. Characterisation of high-altitude Artemia
populations from the Qinghai-Tibet Plateau, PR China. Symposium on Aquatic Biodiversity: Past, Present,
Future, Antwerp, Belgium, 11-13 August 2003.
Other relevant publications and events
Contributions by Gonzalo Gajardo and Patricia Beristain (both ULL) to Seminar on Biodiversity and
Convertation of Animal Genetic Resources (Seminario sobre Biodiversidad y Conservación de Recursos
Genéticos Animales, Santiago de Chile, Chile, 6-8 December 2004.
Consultancy mission report: Status of Artemia exploitation and its potential in Sri-lanka salterns. Aquatic
Resource Development and Quality Improvement Project (ARDQIP). Project Implementation Consultant:
Nguyen Van Hoa, Cantho University, Vietnam
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CONCLUSIONS
This global concerted Artemia study was conceived as a programme of workshops and study visits,
aiming at technical intercalibration, integration of current ecological and evolutionary concepts, and
uniformous methods to assess population dynamics, and as such it has achieved its goals: all
workshops and numerous study visits have been organized and finalized successfully. Intercalibration
of methodologies, and integration of concepts is a continuous process – which is never terminated –
but it is safe to state that this Concerted Action has contributed considerably to the harmonization of
techniques and thoughts among the Consortium members and – through the website, the future
‘Manual’, the attendance of non-Consortium specialists at workshops, and other bilateral and
multilateral contacts – outside the circle of Consortium Artemia specialists. As an example of the latter,
several Artemia experts (ARC contacts) from ex Soviet Union countries have followed the Project
from close, and have been actively involved in its activities (e.g. by oral presentations at Project
workshops; establishment of new research contacts with Consortium members). In a later stage of the
project input was received from Artemia experts in Central Asia and Middle East countries, which
prepared ground for the establishment of a regional consortium of Artemia experts. Within consortium
members, the project gave the opportunity to young aspirants (both in EU and DC partners) for
scientific research. Existing links between partners haven thus been strengthened and new links have
been established. For Consortium members, their participation to this project often facilitated to
generate (financial) support from other authorities, and contributed to a positive evaluation when
applying for related national and international projects.
At the technical and research level, the project has lead to an intensification of contacts and improved
exchange of study material, allowing scientific progress to be made at a faster pace and within a
conceptual framework shared by all Project partners, resulting in the achievement of important
milestones described above (see “Results achieved”). Collaborations and interaction during the three
years of INCO project have endowed Artemia research with significant achievements. The introduction
of molecular systematic analyses and powerful DNA markers has had substantial implications for a)
the study of evolutionary mechanisms in the genus by introducing harmonized methodologies shared
by several research groups in the world, and b) the advancement of aquaculture applications. Scientists
in the field now have common tools and the required expertise to define questions regarding
phylogenetic relationships between species, mechanisms of speciation, models of population structure
and patterns of distribution and invasion. In conjunction with ecologically-related assays used for the
determination of life span and reproductive characteristics and the assessment of phenotypic responses
to various experimental environments, the biodiversity of species, gene pools and habitats (although a
lot of information is still lacking) has been more adequately described and evaluated. Also,
identification of different strains in commercial samples has become a rapid and reliable procedure,
which gives aquaculturists and commercial operations tremendous possibilities for better management,
quality control and biomonitoring of used material. The opportunities offered by the INCO project
have been utilized such, as to provide Artemia research with features sought for in every study:
multidisciplinarity, methodological uniformity and matching of basic and applied research.
With the use of classical tools and molecular tools, this INCO project thus has been instrumental in
unveiling in more detail than before, the biodiversity within the genus Artemia. Though it is clear that
the biodiversity is still incompletely characterized sofar, the data collected within the project should
allow to design experiments that would more completely describe the biodiversity within each species
and within the genus through the application of various genetic techniques on the variety of samples
which are amply available in the resp. members’ cyst banks. For instance, a systematic sequencing
effort on a mitochondrial and a nuclear marker on individuals (including cysts which no longer hatch),
belonging to a well-defined set of samples would even more clearly describe the phylogeny and
phylogeography of the genus Artemia.
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Finally, the Project has been the direct incentive to the realization (even if not finalized) of other
common activities (not provided for in technical annex), like the creation of an integrated cyst bank,
the database of Artemia sites, a website with Artemia-biodiversity related information. Due to the delay
inherent to the publication of research data in scientific literature, the success of this project, quantified
as number of resulting publications, can only be adequately assessed in the near future: lots of research
information are in various states of being processing into manuscripts.
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