LABORATORY OF AQUACULTURE & ARTEMIA REFERENCE CENTER
FACULTY OF AGRICULTURAL AND APPLIED BIOLOGICAL SCIENCES
DEPARTMENT OF ANIMAL PRODUCTION
INCO project Artemia biodiversity
Ghent Workshop, February 5-7, 2002
Rozier 44, B-9000 Gent, Belgium
tel. +32-9-264.37.54 fax +32-9-264.41.93 e-mail: artemia@rug.ac.be
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Programme
Tuesday February 5: Oral presentations
8.30-8.45 Patrick Sorgeloos: Welcome & introduction
8.45-9.05 Gilbert Van Stappen, Belgium (INCO 1)
9.05-9.25 Peter Bossier, Belgium (INCO 2)
Title: A database for Artemia Authentication
9.25-9.45 Elena Boyko, Russia (Guest)
Title: Authentication of Artemia from Russia
9.45-10.05 Theodore Abatzopoulos, Greece (INCO 3)
10.05-10.25 Francisco Amat, Spain (INCO 4)
10.25-11.00 break
11.00-11.20 Graziella Mura, Italy (INCO 5)
11.20-11.40 Godelieve Criel, Belgium (Guest)
Title: Morphological tools to help distinguishing Artemia species and
populations
11.40-12.00 Nguyen Van Hoa, Vietnam (INCO 7)
Title: Artemia pond culture, strain characterisation and its approach in
Vietnam
12.00-12.20 Mohammed Romdhane, Tunisia (INCO 8)
Title : Status of the Artemia Biodiversity in Tunisian salt areas
12.20-13.30 lunch pauze
13.30-13.50 Tom MacRae, Canada (Guest)
Title: Microtubule proteins and chaperones: molecular technologies
applied to study of the brine shrimp, Artemia franciscana
13.50-14.10 Jim Clegg, USA (Guest)
Title: The diversity of Artemia habitats is reflected in the biochemical
repertoire of this remarkable organism
14.10-14.30 Horst Kaiser, South Africa (INCO 9)
Title: Characterisation of southern African Artemia populations, and the
use of Artemia as a vector for probionts and hormones
14.30-14.50 Xin Naihong, China (INCO 10)
14.50-15.10 Naser Agh, Iran (INCO 11)
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15.10-15.50 break
15.50-16.10 Brad Marden, Canada (Guest)
Title: Recovery of the Great Salt Lake, USA, brine shrimp (Artemia
franciscana) population
16.10-16.30 Jorge Castro Mejia, Mexico (INCO 12)
Title: Artemia research in the Universidad Autonoma MetropolitanaXochimilco, Mexico
16.30-16.50 Marcos Camara, Brazil (INCO 13)
Title: Artemia research at Universidade Federal do Rio Grande do
Norte (UFRN), Natal, Brazil
16.50-17.10 Gonzalo Gajardo, Chile (INCO 14)
Title: Artemia characterization, species and speciation
17.10-17.30 Peter Maryan, India (INCO 15)
Title: Research on Artemia in the Institute for Artemia Research and
Training
17.30-17.45 Luc De Meester, Belgium (Guest)
Title: Dispersal, habitat size and genetic differentiation among
zooplankton populations
17.45-18.00 Luc Brendonck, Belgium (Guest)
Title: Adaptations to life in hyper-ilogotrophic ephermeral pools
Wednesday February 6: Discussions
a.m.
General objectives of INCO project; planning of activities
Objectives of stays of visiting scientists
Objectives of regional workshops
p.m.
Discussions of a.m: continued
Project management: financial aspects, reporting
Thursday February 7: Discussions
a.m.
Regional workshops: practical planning
Conclusions and recommendations
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Abstracts - Inco members
INCO partner: 1
Partner name: RUG: Laboratory of Aquaculture & Artemia Reference
Center (ARC), Ghent University, Rozier 44, B-9000 Gent, Belgium
tel. 32-9-264 37 54; fax 32-9-264 41 93; email: gilbert.vanstappen@rug.ac.be
Responsible scientist: Patrick Sorgeloos
Authors: Gilbert Van Stappen, Jean Dhont
Abstract:
The ARC continues the study of various fundamental aspects related to
Artemia biology and its mass-production, e.g. cyst biology and diapause
regulation, strain characterization, filter-feeding kinetics, intensive production
techniques for cyst and biomass, enrichment techniques and HUFA
metabolization in view of application in larviculture. Several collaborative
programmes are running with other INCO partners.
As a consequence of the considerable fluctuations in cyst supply in recent
years, various collaborative projects with foreign universities, local authorities
and/or private partners focus on the population assessment of Artemia
resources, other than Great Salt Lake (esp. in the Central Asian republics and
in Siberia). These ecological studies comprise the study of abiotic parameters,
primary production and Artemia dynamics, and aim at a better understanding
of seasonal and annual fluctuations in brine shrimp densities.
Other research topics are more of a biotechnological nature: a study has been
initiated in cooperation with other Flemish Universities/Departments on the
functional role and characteristics of micro-organisms in the larviculture of
aquatic organisms, with Artemia as a test species. This study aims at
elucidating the pathways of colonization of Artemia by bacteria, at
understanding the infection process by pathogens and finally at developing
and unravelling probiotic methods for countering bacterial diseases. A key
component in this study is the availability of a convenient axenic lab-culture of
Artemia that yields test animals with a ‘normal’ developmental health status
and acceptable growth and survival.
In collaboration with the Laboratory of Microbial Ecology and Technology, the
universities of Dalhousie (Canada) and Connecticut (USA), a proposal has
been submitted to create a Flemish Institute of Biotechnology, “Department of
Microbial Ecology of Aquatic Invertebrates”; focusing on how allochthonous
micro-organisms establish themselves in eukaryote-associated microbial
communities, with Artemia franciscana as the model eukaryote, and a
previously developed culture collection of Vibrionaceae, which are pathogenic
or beneficial to Artemia; as model of allochthonous organisms.
Another line of research is situated in the field of biological pest control, the
culture of insects predating on organisms harmful for crops. Cost-effective
mass rearing systems for these insects should enable the production of large
numbers of insects at the lowest possible price. Preliminary tests have shown
that cysts of Artemia can be used to feed various species of predatory bugs,
be it with variable success, as an alternative to the expensive lepidopteran
eggs, routinely used. Future study will determine the developmental and
reproductive characteristics of a number of insects, when fed Artemia cysts
supplied in different forms.
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Relevant publications:
Dhont, J. & Sorgeloos, P. in press. Applications of Artemia. In “Th.
Abatzopoulos, J. A. Beard-more, J. S. Clegg & P. Sorgeloos (eds) “Artemia:
basic and applied biology”. Kluwer Academic Publishers, Dordrecht,
Netherlands
Han, K., Geurden, I. & Sorgeloos, P. 2001. Fatty acid changes in enriched
and subsequently starved Artemia franciscana nauplii enriched with different
essential fatty acids. Aquaculture 199: 93-105
Lavens, P. & Sorgeloos, P. 2000. The history, present status and prospects of
the availability of Artemia cysts for aquaculture. Aquaculture, 181: 397-403
Sorgeloos, P., Dhert, P. & Candreva, P.2001
Use of the brine shrimp, Artemia spp., in marine fish larvicultur. Aquaculture,
200: 147-159
Van Stappen, G. in press. Zoogeography. In “Th. Abatzopoulos, J. A.
Beardmore, J. S. Clegg & P. Sorgeloos (eds) “Artemia: basic and applied
biology”. Kluwer Academic Publishers, Dordrecht, Netherlands
Verschuere, L., Heang, H., Criel, G., Sorgeloos, P. & Verstraete, W. 2000.
Selected bacterial strains protect Artemia spp. from the pathogenic effects of
Vibrio proteolyticus CW8T2
Applied and environmental microbiology, 66(3):1139-1146
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INCO partner: 2
Partner
name:
CLO-DVZ:
Agricultural
Research
CenterGhent/Department of Sea Fisheries (CLO-DVZ), Ankerstraat 1, 8400
Oostende, Belgium
tel. 32-9-34 22 66; fax: 32-9-33 06 29; email: pbossier@yucom.be
Responsible scientist: Peter Bossier
Authors: Francesco Catania, Fanny Dooms, Gilbert Van Stappen, Patrick
Sorgeloos, Eddy Naessens, Peter Bossier
Title: A database for Artemia authentication.
Abstract: A database has been constructed for the authentication of Artemia
strains. The authentication is based on the RFLP analysis of a 1300 bp
mitochondrial ribosomal DNA fragment, using six restriction enzymes. In total
38 samples have been analysed. The currently described species could be
easily distinguished except for A. urmiana and A. tibetiana. Within the species
A. franciscana, cysts originating from GSL could be distinguished from those
originating from SFB. There are indications that GSL contains two genotypes.
In total 18 A. parthenogenetica strains have been analysed. The RFLP
dendrogram indicates that there are at least 4 clusters within the A.
parthenogenetica species.
Relevant publications:
Camargo, W., Sorgeloos, P. & Bossier P. 2002 Preliminary genetic data on
some Caribbean Artemia franciscana strains based on RAPD's. Int. J. Salt
Lake Res. In press
Abatzopoulos, T.J., Kappas, I., Bossier, P., Sorgeloos, P. & Beardmore, J.A.
Genetic characterisation of Artemia tibetiana (CRUSTACEA, ANOSTRACA).
submitted.
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INCO partner: 3
Partner name: AUTH: Aristotle University of Thessaloniki, Department of
Genetics, Development & Molecular Biology, Faculty of Sciences,
School of Biology, 540 06 Thessaloniki, Greece
tel.: 30-31-099 83 01; fax. 30-31-099 82 56; email: abatzop@bio.auth.gr
Responsible Scientist: Theodore J. Abatzopoulos
Authors: Theodore J. Abatzopoulos, Th. Baxevanis
Abstract:
Experience of the team:
This team has been working on the genetic structure and characterization of
Artemia populations (and other species) for the last 18 years. Expertise is
established mainly on the following aspects:
 Population structure by using molecular markers (RFLPs, RAPDs,
AFLPs)
 Chromosomes, karyotypes and chromocentres
 Protein analyses i.e. allozyme polymorphism, stress proteins, by using
electrophoretic approaches (starch-gel, SDS-PAGE, isoelectric
focusing)
 Biometry and morphology
 Effects of temperature and salinity on reproductive and life-span
characteristics
 Clonal diversity in parthenogenetic populations
 Mechanisms of diapause deactivation
 Field work at nearby saltworks
Role and contribution of AUTH to this project:
This group could be responsible for the genetic characterization of different
Artemia populations, bisexual and parthenogenetic, according to their mode of
reproduction, ploidy level, life-history traits and clonal diversity by focusing on
multidisciplinary approaches (for methodologies see above).
Future objectives:
 Population/strain characterization by using molecular markers
 Clonal diversity in parthenogenetic Artemia
 Diapause deactivation mechanisms (focusing on genetics)
Relevant publications:
Abatzopoulos, T.J., G.V. Triantaphyllidis, J.A. Beardmore & P. Sorgeloos.
1997. Cyst membrane protein composition as a discriminant character in the
genus Artemia. (International study on Artemia LV). J. Mar. Biol. Ass. UK 77:
265-268.
Triantaphyllidis, G.V., G.R.J. Criel, Th.J. Abatzopoulos, K.M. Thomas, J.
Peleman, J.A. Beardmore & P. Sorgeloos.1997. International Study on
Artemia. LVII. Morphological and molecular characters suggest conspecificity
of all bisexual European and North African Artemia populations. Marine
Bioogy. 129: 477-487.
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Abatzopoulos, Th.J., B. Zhang & P. Sorgeloos. 1998. Artemia tibetiana:
preliminary characterization of a new Artemia species found in Tibet (People’s
Republic of China). International Study on Artemia. LIX. Int. J. Salt Lake Res.
7: 41-44.
Triantaphyllidis, G.V., Th.J. Abatzopoulos & P. Sorgeloos. 1988. Review of
the biogeography of the genus Artemia (Crustacea, Anostraca). J. Biogeogr.
25: 213-226.
Abatzopoulos, Th.J., I. Kappas, P. Bossier, P. Sorgeloos & J.A. Beardmore.
2002. Genetic characterisation of Artemia tibetiana (CRUSTACEA,
ANOSTRACA). Biological J. Linnean Soc. (in press)
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INCO partner: 4
Partner name: CSIC: Instituto de Acuicultura de Torre de la Sal (Consejo
Superior de Investigaciones Científicas de España), Larval Food
(Artemia), Larviculture and Ecotoxicology group, 12595 Ribera de
Cabanes (Castellón), Spain
tel.: 34-964-319500; fax: 34-964-319509; email: amat@iats.csic.es
Responsible scientist: Francisco Amat
Author: Francisco Amat
Abstract:
For the last three years, and during the next three years, according to the
commitments acquired in our research projects, we were concerned and we
are going to develop the following fields and topics related to Artemia study:
1. Biodiversity of Artemia species around the world, especially from South
America, and more specifically
from Argentina (International AECI
poject/Spain-Argentina) aiming to establish the distribution of A. persimilis and
A. franciscana populations and their morphological differentiation through
cysts, nauplii and adult specimens biometrics. Their genetic and reproductive
isolation (intraspecific and interspecific crosses), together with reproductive
aspects in terms of the factors driving oviparism versus viviparism and quality
of the offspring, specially in cyst hatching. Application of all these aspects to
competition studies in laboratory as well as in outdoor conditions (National
I+D+i Programme project).
2. Factors conditioning PUFA levels in cysts, nauplii and biomass from
different Artemia (autochthonous and foreign) populations, related to their
origin and ecological conditions, specially according to the type and quality of
food available (microalgae, halobacteria, etc). The importance of these levels
for marine fish larviculture (seabass, seabream, common dentex) and the
possibility to manipulate and improve these PUFA, as well as other nutrient,
levels through enrichments with lipid emulsions and liposomes (National I+D+i
Programme P4 project).
3. Application of Artemia to aquatic ecotoxicological studies (heavy metals
and pesticides) aiming to use Artemia as a target species or as a tool to
vehiculate xenobiotics to early developmental stages of fishes and shrimps, in
freshwater and seawater. A research line on bioaccumulation and
biodepuration processes, and study of physiological evidences of these
mechanisms through biomarkers is under development.
4. Related to the study of the different aspects formerly cited dealing with
Artemia ecology formerly cited, a research line on Artemia biology affected by
parasitism (Cestoda, Hymenolepidida), is being developed in autochthonous
populations. Very interesting phenomena related to population development
through reproduction mode conditioning are being unveiled.
The development of the INCO project commitments that the Inst. de Acuicult.
de Torre de la Sal group acquired with the other EU groups, as well as with
the non-EU groups through the participation of their members in workshops
and training courses, with the application of their own and autochthonous
Artemia species and populations, bringing adequate cysts samples, should
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contribute to attain the success of the INCO project in the different topics
stated in the very proposal, i.e: to improve on general topics of species
description and characterization; populations identification, biodiversity and
intercalibration of methodologies aiming, finally, to a better knowledge of this
natural resource for a better and reasonable aquaculture.
Relevant publications:
Zuñiga, R., R. Wilson, F. Amat & F. Hontoria. 1998. Distribution and
characterization of Chilean
populations of the brine shrimp Artemia
(Crustacea, Branchiopoda, Anostraca). Int. Journal of Salt Lake Research. 8
(1): 23-40.
Cohen, R.G., F. Amat, F. Hontoria & J.C.Navarro. 1999. Preliminary
characterization of some Argentinean Artemia populations from La Pampa
and Buenos Aires provinces. Int. Journal of Salt Lake Research. 8 (4): 329340.
Navarro, J.C., L.A. McEvoy, R.J. Henderson & F. Amat. 1999. Lipid
conversions during Artemia enrichment. Aquaculture 174: 155-166.
Varó. I., A.C. Taylor, J.C. Navarro & F. Amat. 2000. Effect of parasitism on the
respiration rates of adults of different Artemia strains from Spain. Parasitology
Research. 86(9): 772-774.
Varó. I., R. Serrano, E. Pitarch, F. Amat, F.J. López & J.C. Navarro. 2000.
Toxicity and concentration of chlorpyrifos in aquatic organisms: Artemia
parthenogenetica (Crustacea), Gambussia affinis and Aphanius iberus
(Pisces). Bull.Environm.Contam. Toxicol. 65:623-630.
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INCO partner: 5
Partner name: LA SAPIENZA: Dipartamento di Biologia Animale é
dell’Uomo, Laboratorio di Zoologia Applicata, Università La Sapienza,
Viale dell’ Università, 00185 Rome, Italy
tel.: 39-6-49914769; fax: 39-6-4958259; email: graziella.mura@uniroma1.it
Responsible scientist: Graziella Mura
Author: Graziella Mura
Abstract:
The Laboratory of Applied Zoology, Dept. Animal and Human Biology, Rome
La Sapienza, has a long tradition in the study of Crustacea Anostraca.
Besides systematics, all the main aspects of biology and ecology are covered.
Recent research has focused on two main subjects:
1. Adaptive strategies and dormancy (hatching phenology, intraspecific
variation in relation to habitat characteristics, genetic and epigenetic origin of
variation of cyst hatching behaviour.
2. Zoogeography and biodiversity. Within this framework, attention is also
given to brine shrimp and brackish water fairy shrimp populations from the
Mediterranean, and Italy in particular. As to Artemia, Italian populations were
monitored and studied from a morphological point of view. Their life history
was investigated in the field, also in relation to the presence of Flamingolepis
parasites.
Bisexual populations from Sardinia were also characterized from a genetic
point of view in cooperation with the team of Prof. Barigozzi, (Milano
University, Dept. Genetics and Molecular Biology). Finally, the applied aspects
were considered.
Present research includes three main aspects: 1) characterization, nutritional
value and productivity of the populations living in the Italian salterns and
coastal ponds (the results of a study on the population from Tarquinia are in
preparation), 2) impact of UV radiation on hatching characteristics (in coll. with
the Institute of Physics, La Sapienza, Prof. Maurizio Severini), and 3) the role
of Artemia as a bioindicator of environmental contamination by trace elements
(in coll. with the Dept. Of Environmental Chemistry, Rome La Sapienza, Prof.
Bianca Petronio)
Relevant publications:
Mura, G. 1990. Artemia salina from Lymington: frontal knob morphology by
means of SEM. Journal of Crustacean Biology, 10(2): 364-368.
Mura, G. 1993. Seasonal distribution of Artemia salina and Branchinella
spinosa in a saline astatic pond of South West Sardinia (Italy). Crustaceana,
64 (2): 172-191.
Mura, G. 1995. Ecological study of a bisexual Artemia population from
Sant'Antioco solar saltworks (south west Sardinia, Italy). International Journal
of Salt Lakes Research 3: 201-219.
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Mura, G. 1995. A case of Cestode parasitism (Flamingolepis liguloides
Gervais, 1847 Spassky & Spasskaja 1954) affecting an Artemia population
from SW Sardinia. International Journal of Salt Lake Research 3: 191-200.
Mura, G. 2001. Updating Anostraca (Crustacea, Branchiopoda) distribution in
Italy. J. Limnology, 60 (1):45-49.
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INCO partner: 7
Partner name: CTU: Institute of Science for Aquaculture, 3rd February
Street, Campus II, Can Tho University, Can Tho, Vietnam
tel.: 84-71-834307; fax: 84-71-830323; email: nvhoa@ctu.edu.vn;
nvhoa.ct@bdvn.vnd.net
Responsible scientist: Nguyen Van Hoa
Author: Nguyen Van Hoa
Title: Artemia pond culture, strain characterization and its approach in
Vietnam
Abstract:
The first introductions of Artemia in Vietnam were done in 1982 in Hon Khoi
(Nha Trang) and later in Qui Nhon, Cam Ranh and Bac Lieu (1983). A number
of Artemia strains have been used to inoculate, e.g. Macau, GSL and China in
Nha Trang (1987) while SFB and Tientsin were attempted in Vinh Chau and
Bac Lieu (1984-1985). However, large-scale cyst production mainly expanded
in Vinh Chau and Bac Lieu, although successful cyst production was soon
reported from other areas. By then, pond culture techniques had become a
main interest of the project and different approaches (e.g. culture systems,
pond management procedures etc.) have been tried in order to achieve an
improved and sustainable cyst production. Besides, an extension program has
been developed to support local salt farmers (i.e. training, technical advice
etc.) to realize large-scale production. Thanks to this program, through which
not only poor farmers could benefit, researchers found out that a big variation
of cyst production occurred from pond to pond, from scale to scale and from
location to location; this topic still remains to be resolved. On top of that the
pond environment seems to be an important factor as any variation of
environmental factors, especially if exceeding the limits of the Artemia
species, will lead to a failure of pond production.
Recently, an Artemia project has been implemented between a number of
laboratories (1) involved to study the adaptation and microevolution of Artemia
franciscana (USA) in the Vinh Chau and Bac Lieu salt fields. SFB was
inoculated in Vinh Chau and its offspring, in terms of cysts, has been used to
study the phenotye and genotype of their genealogy. Primary results indicate
that the phenotype has varied according to the culture environment, but that
also the genotypes (i.e. allozymes, mtDNA, heat-shock proteins) show a
detectable genetic differentiation between the initial SFB and its offspring, and
especially between SFB and VC, which has been established more than 10
years in the site of Vinh Chau, and which originated from SFB. There is
scattered evidence for a correlation between individual heterozygosity, and
there is evidence for differences in heat resistance between VC and SFB.
There is no appreciable difference in the degree of hsp 70 up-regulation
between SFB and VC. Within a number of years/generations, the project has
generated cyst samples from SFB until Y3; it is however possible that genetic
differentiation is not yet shown or that other approaches in the study of this
subject are needed. Therefore, a continuation of recruitment of the SFB (here
SFB 1258) in the site and different approaches are strongly recommended for
the project.
(1) Artemia Reference Center, Ghent University, Gent, Belgium; Bodega
Marine Laboratory, University of California (Davis), Bodega Bay, USA;
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School of Biological Sciences, University of Wales, Swansea and
Aquaclture and Fisheries Sciences Institute, Can Tho University, Vietnam.
Relevant publications:
Anh, N.T.N., Quynh, V.D., Hoa, N.V & Baert, P. 1995. Potential for Artemia
biomass production in Vinh Chau salterns. The First Symposium on Marine
Biology, Oct. 27-28, 1995, Oceanography Institute, Nha Trang, Vietnam
(Abstract in English).
Anh, N.T.N., Quynh V.D., Hoa N.V & Baert, P. 1997. Present situation of
Artemia and salt production in the coastal salinas from Soc Trang and Bac
Lieu provinces. The First Science Symposium, Can Tho University, Vietnam
(Abstract in English).
Baert, P., Anh, N.T.N., Quynh, V. D & Hoa, N. V. 1997. Increasing cyst yields
in Artemia culture ponds in Vietnam: the multi-cycle system. Aquaculture
Research, (28), 809-814.
Clegg, J.S., Jackson, S.A., Hoa N.V. & Sorgeloos, P. 2002. Comparison of
thermal resistance, developmental rate and heat shock proteins in brine
shrimp, Artemia franciscana, from San Francisco Bay and southern Vietnam.
(Hydrobiologa, in press).
Vu Do Quynh & Nguyen N. L.. 1987. Inoculation of Artemia in experimental
ponds in central Vietnam: an ecological approach and a comparison of three
geographical strains. In: Artemia Research and its Applications. Vol. 3.
Ecology, Culturing, Use in Aquaculture. P. Sorgeloos, D.A. Bengtson, W.
Decleir & E. Jaspers (Eds). Universa Press, Wetteren, Belgium. 253-269.
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INCO partner: 8
Partner name: INAT: Institut National Agronomique de Tunisie,
Université de Carthage, 43 av. Charles Nicolle 1082, Tunis, Tunisia
tel. :
216-1-287110;
fax:
216-1-799391;
email:
romdhane.medsalah@inat.agrinet.tn
Responsible scientist: Mohamed Salah Romdhane
Authors: Romdhane M.S., Ghlala A.
Title: Status of the Artemia biodiversity in Tunisian salt areas
Abstract:
Actually four representative populations of Artemia are identified and sampled
from Tunisian sebkhas and salt works; Chott Jerid (southern inland area),
Salina of Sfax (Southern coastal area), Sebkha Corsia (northern inland area)
and Sebkha Sijoumi (northern coastal area).
Morphologic and reproductive characteristics are analysed from field samples
and laboratory culture.
A total of eight biometric analyses are established with total length, antenna,
furca, and brood sac sizes, where two morphometric groups are distinguished.
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INCO partner: 9
Partner name: RU: Department of Ichthyology and Fisheries Science, PO
Box 94, Rhodes University, Prince Alfred Street, Grahamstown 6140,
South Africa
tel.: 27-46-6038415 ; fax : 27-46-6224627 ; e-mail: h.kaiser@ru.ac.za;
t.hecht@ru.ac.za
Responsible scientist: Tom Hecht
Authors: Horst Kaiser, Tom Hecht
Title: Characterization of southern African Artemia populations, and the use of
Artemia as a vector for probionts and hormones
Abstract:
1) Genotype/environment interactions in Southern African Artemia populations
Artemia populations from the southern African region have been reported for
Namibia, Mozambique and Botswana and these populations need to be
characterized and their distribution established. There is a paucity of data on
isolated populations, their distribution and differences between them.
Genotype/environment interactions are worth investigating to test how these
populations differ from each other under culture conditions. Aspects, such as
temperature tolerance and rate of energy loss after hatching need to be
investigated. Cysts will have to be collected from different sources, and the
performance of the nauplii compared with each other. Factors to be tested are
hatching characteristics, and chemical composition, for example fatty acid
profiles. Also, in conjunction with the other projects listed in this abstract,
population-specific ability of nauplii to act as a vector for biochemical
compounds and probiotic bacteria should be investigated.
2) Use of Artemia as a vector in the application of probiotic bacteria
Typical bacterial concentrations in coastal waters range from 105 to 107 cells
per ml and larval survival in marine fish hatcheries may be affected not only
by the number of bacteria in the water but also the species composition. The
application of probiotic bacteria could be a way to introduce bacteria that can
compete with pathogenic bacteria and consequently improve larval survival.
Results from laboratory in vitro studies in our department suggest the
existence of various probiotic strains that effectively compete with fish-specific
pathogenic bacteria. Introducing such bacteria via live food could make them
available to fish larvae.
The bacterial load of rotifers has been shown to range from 10 4 to 105 per
individual and it corresponds well with the composition of the bacteria in the
culture water, thus indicating a non-selective attachment. To investigate if
Artemia are suitable vectors for probionts we suggest a series of experiments.
These include the testing of different conditions and application of varying
quantities of probionts to Artemia. We aim to test the possibility of adding
probionts pre-hatch after decapsulation, and adding them immediately prior to
feeding and to check attachment using SEM techniques. We will also aim to
test the duration of probiont viability in Artemia culture, and the potential effect
of probiotic bacteria on the health and viability of the Artemia. Strains have
been identified and, using PCR techniques, we can make and use primers to
quantify attachment.
3) Artemia as a carrier in the application of GTH releasing hormone
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Pilot experiments conducted in our department showed the potential of
delivering a liquid gonadotropin hormone-releasing synthetic decapeptide via
Artemia to induce ovulation in a small characin species (Cardinal Tetra,
Paracheirodon axelrodi). The data suggest that there would be an optimal
time of immersion of Artemia in the hormone solution and that this could be an
effective way to administer spawning hormones to small, valuable species,
where intramuscular injection is not practical. Prolonged immersion decreases
efficacy, possibly because the hormone is broken down. When the duration of
immersion was too short the hormone was probably not picked up in sufficient
quantities.
We need to investigate this further as it is not understood how the hormone
has been picked up, how fast it can get broken down in the Artemia, and how
we can determine the optimal period of immersion. It would also be important
to test the use of encapsulated hormones.
Relevant publications:
Burton, S., H. Kaiser & T. Hecht. 1998. The potential of Artemia-mediated
delivery of a gonadotropin hormone analogue to induce ovulation in the
cardinal tetra (Paracheirodon axelrodi).
Aquarium Sciences and
Conservation, 2, 89-92.
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INCO partner: 10
Partner name: SRI: Salt Research Institute, Yingkou Road 831, 300450
Tanggu, Tianjin, PR China
tel.: 86-22-25301094; fax: 86-22-25301094; email: srisalt@public.trp.tj.cn
Responsible scientist: Xin Naihong
Author: Xin Naihong
Abstract :
The Salt Research Institute (SRI) started the research on brine biotechnology
through the UNDP project “Biotechnology in solar salt fields” in 1987. This
early study mainly focused on the biological management of the solar
saltworks in the Bohai Bay and the utilization of Artemia cysts and biomass in
local aquaculture. In 1991 a joint research project with ARC (coordinator) and
the Universities of Wales, Swansea (UK) and Milan (Italy) was initiated with
funding from the European Union. The main focus of this joint project was the
identification and characterization of Chinese Artemia strains for use in
aquaculture. This project was very successful as important contributions were
made to the fundamental knowledge of Artemia biology, ecology, as well as in
relation to the selection and use of specific brine shrimp strains for use in
aquaculture. The Artemia franciscana introductions in solar salt production
have succeeded through the VLIR-ABOS project “Development and
application of new techniques for improved Artemia, shrimp and salt
production in PR China” (1993-1996). The food value of various local Artemia
sources in larviculture of shrimp and marine fish have been tested in this
project. From 1996, continuous studies mainly focus on the specific areas with
special characteristics (Tibet, Inner Mongolia) through 2 joint projects, entitled
‘Further exploration, characterization and utilization of the natural brine shrimp
Artemia resources in the People’s Republic of China’ (1996-1999) and ‘Study
of the biodiversity of Chinese Artemia strains and their possible application in
research and aquaculture’(2000-2001).
Research themes:
1. Artemia resources in China
2. Strain characterization and nutrition of Chinese Artemia
3. Introduction and culture of Artemia in China
4. Artemia application in China.
Expectations from INCO project:
1. Uniformity of systematic classification for Chinese Artemia strains
2. Continuous studies on the new Chinese species
3. Uniformity of methodologies for study of Artemia
4. Suggestions on sustainable development of Artemia resources
Relevant publications:
Xin, N., Sun, J., Zhang, B., Triantaphyllidis, G.V., Van Stappen, G. &
Sorgeloos, P. 1994. International Study on Artemia. LI. New survey of Artemia
resources in the People’s Republic of China. International Journal of Salt Lake
Research, 3: 1-8.
17
Xin, N., Sun, J., Guizheng, Z. & Sorgeloos, P. 1999. The effect of Artemia
sources on growth, survival and salinity stress of crab (Eriocheir sinensis)
larvae. Asian Fisheries Science 12:201-205.
Xin, N., Audenaert, E., Vanoverbeke, J., Brendonck, L., De Meester, L. &
Sorgeloos, P. 2000. Low among-population genetic differentiation in Chinese
bisexual Artemia populations. Heredity 84: 238-243.
Zhang, B., Xin, N., Yu, X. & Sui, L. 1998. Strain characterization of Artemia
from 9 inland salt lakes of China. Journal of Lake Sciences, Vol. 10, No. 3: 1923.
18
INCO partner: 11
Partner name: UU: Artemia and Aquatic Animals Research Center
(AAARC), Urmia University, 165 Shahid Beheshty Av., 57153, Urmia, Iran
tel: 98-441-440295; fax: 98-441-440295; email: agh1960@hotmail.com
Responsible scientist: Naser Agh
Author: Naser Agh
Abstract:
The author has been involved in Artemia research activities since 1992.
Artemia work was started with research on the biology of A. urmiana and its
post-embryonic development. Present research focuses on its biology and
characterization, and on the co-existence of bisexual and parthenogenetic
strains and the identification of new populations in Iran. The author is
coordinating a number of research activities at AAARC:
1. Characterization of Artemia populations from Iran:
Artemia has been reported from about 10 different geographical locations
from Iran, but there were no scientific data about them so far, except for A.
urmiana from Urmia Lake. During this study cyst samples were collected from
7 different locations and their life cycle and morphometric charateristics were
studied, which revealed that the Artemia existing in different lakes and
lagoons in Iran (except Urmia Lake) are all parthenogenetic populations.
Noteworthy is the presence of a parthenogenetic population just at the vicinity
of Urmia Lake.
In order to study the Artemia from Urmia Lake more closely, cysts were
collected from different sites of the lake and the newly hatched larvae were
cultured as a function of different salinities, starting from 15 ppt till 250 ppt. At
low salinities almost always some parthenogenetic Artemia were found, and at
low salinities the bisexual population never attained sexual maturity, but the
parthenogenetic population grew into adult females, which reproduced. This
explains why A. parthenogenetica exists in the lagoons, which initially (in the
beginning of spring) have very low salinity, but bisexuals do not. On the other
hand it was observed that when the newly hatched larvae were cultured at
higher salinities, rarely any parthenogenetic individuals were observed; this in
turn explains why the density of the parthenogenetic females is negligible in
the lake itself.
2. Effect of anti-bacterial and anti-fungal agents on hatching quality of
contaminated A. urmiana cysts:
In this study different anti-bacterial treatments were used, such as antibiotics,
heat shock and UV irradiation, and different anti-fungal agents such as
formalin, malachite green, and methylene blue in different doses, and these
were compared for their efficiency in improving hatching quality. The results
were compared with routine hypochlorite disinfection and decapsulation of
cysts. It was observed that antibiotics and formalin improved the hatching
percentage by about 15%, but decapsulation still proved to be a better
method, as it promoted H% by 20%.
3. Effects of laser, gamma and beta irradiations:
In this study we are investigating the effects of Helium-Neon laser, Gamma
and Beta irradiations on the hatching quality of cysts and on the life cycle
characteristics of A. urmiana.
19
4. Pond culture of Artemia:
Iran has a high potential for the pond culture of Artemia. There are a number
of inland saline lakes, lagoons and even rivers in different parts of Iran.
Moreover there are hundreds of thousands of hectares of land at the southern
border of Iran in the vicinity of the Persian Gulf and the Oman Sea, where
salinity varies between 35-45 ppt. These areas provide best opportunity for
the pond culture of Artemia. This possibility was tested by inoculating A.
franciscana (Vietnam strain) at Khozestan province, which gave very
promising results. Therefore extensive pond culture projects are organized,
which will begin from March 2002 onwards. An estimated 20 metric tons of
good quality cysts will be produced by mid November 2002. This will certainly
help further development of fish and shrimp culture in Iran and moreover it will
improve the economy of the local people who will be involved in this activity in
different provinces.
5. Ecological study of Urmia Lake and Artemia resource assessment:
Urmia Lake is facing one the most severe ecological crises ever seen.
Presently the lake water is saturated with brine. Its salinity is about 300 ppt
and salt precipation is observed everywhere around the lake. There is a thick
layer of sedimented salt on the lake bottom. All this affects the Artemia
population, the dominating creature in the lake. The production rate has
greatly decreased and the Artemia resources have reached the lowest level
ever. Thousands of migrating and local birds died due to the absence of food
and salt toxicity. This situation requires a team of experts to work on this
unique ecosystem with special emphasis on saving A. urmiana from this
crisis.
A project, entitled “Ecological study of Urmia Lake and Artemia resource
assessment” has been proposed to the Environmental Protection
Organization of the Islamic Republic of Iran, and has been approved. This two
year-project, which is supposed to start from January 2002 onwards, will
receive assistance from ARC, Gent. During this study the ecological changes
at different sites and depths will be recorded, and through a sampling
campaign the Artemia and phytoplankton population will be assessed
qualitatively and quantitatively.
Expectations from INCO Project:
I expect the INCO Project to help us in a more detailed study on the
characterization of Artemia populations from the Middle East and Central
Asian countries. We also expect the INCO project to assist us to get more
experienced in pond culture of Artemia.
20
INCO partner: 12
Partner name: UMA-X: Universidad Autónoma Metropolitana-Xochimilco,
Depto. El Hombre y su Ambiente, Calz. Del Hueso No. 1100. Col. Villa
Quietud. México, 04960, D.F, Mexico
tel.: 52-5-4837151; fax: 52-5-4837469; email: cabt7515@cueyatl.uam.mx
Responsible scientist: Talía Castro Barrera
Authors: Talía Castro Barrera, Jorge Castro Mejía, Germán Castro Mejía,
Ramón de Lara Andrade, Aida Malpica Sánchez
Title: Artemia research in the Universidad Autónoma MetropolitanaXochimilco, Mexico
Abstract:
Since 1979, the group of the Universidad Autónoma Metropolitana-Xochimilco
has been working with this crustacean. At present, 17 places are known with
natural populations of Artemia, of which three are located in inland waters, two
populations in the Gulf of Mexico, one in the Caribbean Sea and the
remaining ones, 11 in coastal waters of the Ocean Pacific. As for the places
where this crustacean is located, we have ecological information on their
climate, but only for a few on the chemical composition of the water and salt.
As biological characteristics, the generated information is: a) morphometry of
cysts, nauplii and female and male adults; b) quantity of nauplii, cysts and
immature eggs produced per female; c) electrophoretic patterns; d)
reproductive isolation and e) biochemical composition (fatty acids and amino
acids patterns of nauplii (24 h) and biomass). We have installations to make
cultivation tests to obtain biomass and use it to feed fishes and crustaceans.
Also, we have gas chromatography, amino acids analyzer and atomic
spectrophometry equipments.
Relevant publications:
Castro, B. T., De Lara, A. R. y Castro, M. J. 1994. El crustáceo Artemia
franciscana alimentado con Spirulina spp. fresca, como dieta de especies
acuáticas comerciales. pp.15-20. En: Hidrobiológica, 4(1-2). UAM-Iztapalapa.
Castro, M. G., Castro, M. J., Castro, B. T., De Lara, A. R. y Malpica, S. A.
1996. Evaluación de la composición y densidad de la Artemia franciscana en
la salina “Tres Hermanos”, Yavaros, Sonora. pp. 127-136. En: Oceanología.
Vol. 1(9).
Castro, B.T., Castro, M.G.., Castro, M.J., Malpica, S.A. y De Lara, A.R. 1997.
Características Morfométricas y Calidad de los Quistes de Artemia sp
(Crustacea: Anostraca), Habitante de Aguas Sulfatadas de Coahuila, México.
En: Ciencias Marinas.
Castro, T., Málpica, A., Castro, J., Castro, G. y De Lara, R. 2000.
Environmental and biological characteristics of Artemia ecosystems in Mexico:
An updated review. 121 – 201. In: M. Munawar, S. G. Lawrence, I. F.
Munawar and D. F.Malley (Eds.) Aquatic ecosistems of Mexico: Status and
Scope, Ecovision World Monograph Series. Backhuys Publishers, Leiden,
The Netherlands. 435 p.
21
Castro, M.G., Malpica, S.A., De Lara, A.R., Castro, M.J. y Castro, B.T. 2001.
Técnicas de cultivo de especies planctónicas e invertebrados útiles para la
acuicultura. Serie Académicos. UAM-Xochimilco. División de Ciencias
Biológicas y de la Salud. 65 p.
22
INCO partner: 13
Partner name: UFRN: Departamento de Oceanografia e Limnologia,
Universidade Federal do Rio Grande do Norte, Campus Universitario, BR
101, 59072-970 Natal, Brazil
tel.: 55-84-2154433; fax: 55-84-6421815;email: mrcamara@ufrnet.br
Responsible scientist: Marcos R. Camara
Author: Marcos R. Camara
Title: Artemia research at Universidade Federal do Rio Grande do Norte
(UFRN), Natal, Brazil
Abstract:
Artemia franciscana has become established in the State of Rio Grande do
Norte (RN) in Northeastern Brazil as a result of inoculations made in Macau in
April 1977 with cysts from a San Francisco Bay (California, U. S. A.) stock.
Universidade Federal do Rio Grande do Norte (UFRN) has been involved in
Artemia studies in RN for about two decades. Major Artemia research
interests at UFRN relate to the reproductive characterization of different
populations of Artemia, the transplantation/inoculation of Artemia strains for
enhanced aquaculture production, and the development of Artemia pond
culture techniques. In this context, UFRN, in partnership with the Brazilian
Shrimp Farmers Association (ABCC), Brazilian Mariculture Linkage Program
(BMLP), and BioArtemia Ltd., runs a pilot farm for Artemia research
(reproductive biology, pond ecology, and aquaculture techniques) in Grossos
(RN) since May 2001. Further areas of interest to broaden Artemia R & D
activities at UFRN include genetic and biochemical characterization of Artemia
franciscana populations in RN.
Relevant publications :
Camara, M. R. 2001. Dispersal of Artemia franciscana (Crustacea: Anostraca:
Artemiidae) populations in the coastal saltworks of Rio Grande do Norte,
Northeastern Brazil. Hydrobiologia: in press
Camara, M. R. 2001. Arachidonic acid: a biological indicator of physiological
stress in the brine shrimp Artemia. In: LARVI 2001 – 3rd FISH & SHELLFISH
LARVICULTURE SYMPOSIUM, 2001, Gent. European Aquaculture Society
Special Publication. Oostende, v.30. p.100-103.
Monteiro, P. A. B. C. & Camara, M. R. 2001. Estruturação e
operacionalização de uma unidade piloto para a produção de cistos e
biomassa de Artemia em regime de cultivo semi-intensivo. Revista da ABCC:
in press.
Camara, M. R. 2000. Artemia no Brasil: do extrativismo ao cultivo. Panorama
da Aqüicultura, 10(62): 15-19.
Camara, M. R. & Pena, O. M. 2000. Reproductive performance of inoculated
Artemia franciscana in a coastal saltwork in NE-Brazil. In: AQUA 2000 RESPONSIBLE AQUACULTURE IN THE NEW MILLENNIUM, 2000, Nice.
European Aquaculture Society Special Publication. Gent, v.28. p.107-108.
23
INCO partner: 14
Partner name: ULL: Department of Basic Sciences/Laboratory of
Genetics & Aquaculture, Universidad de Los Lagos, P.O. Box 933,
Avenida Fuchslocher s/n, Osorno, Chile
tel. 56-64-205293; fax: 56-64-239517; email: ggajardo@ulagos.cl
Responsible scientist: Gonzalo Gajardo
Author: Gonzalo Gajardo
Title: Artemia characterization, species and speciation
Abstract:
The speciation process is a relevant, though still poorly known, aspect of
evolutionary theory, and critical to understand the origin and maintenance of
biodiversity. Characterisation studies on Artemia have provided so far a
significant database (field and laboratory data) at different levels, i.e.
morphology, karyology, physiology, nuclear and extra-nuclear DNA, which are
relevant to understanding how recombination and adaptive release of
genotypic and phenotypic variation affect the speciation process in Artemia.
There is a need though to integrate all these results into a coherent body, and
in particular to rethink the validity of the Biological Species Concept in
Artemia, a genus with sexual and asexual reproduction. Likewise, to reconcile
the information obtained from genotypic and phenotypic spaces, two
dimensions that are co-adapted (Gajardo and Beardmore, 2001), i.e.
integrated via specific regulatory interactions at every level of the individual’s
hierarchical organisation.
In this work, key questions concerning species and speciation are deduced
from the data available in Artemia, a genus suited with a range of evolutionary
tools not very commonly seen at once in other organisms. In particular: i) how
morphological discontinuities known as species can be produced from a
continuous process (evolution); ii) importance of the use of different traits,
either singly or combined, to identify Artemia species and to providing a more
realistic view of the diversity of the genus. What reveal inter and intra-specific
comparisons; iii) the Biological Species Concept and the need to integrate two
gene pools in Artemia (the sexual and asexual); iv) Testing the allopatric
mode of speciation.
Relevant publications:
Gajardo, G., Mercado, C. Beardmore, J.A. & Sorgeloos, P. 1999. International
study on Artemia. LX. Allozyme data suggest that a new Artemia population in
southern Chile (50º29’S; 73º45’W) is A. persimilis.. Hydrobiologia. 405: 117123.
Gajardo, G., Parraguez, P. & Beardmore J.A. & Sorgeloos, P. 2001.
Reproduction in the brine shrimp Artemia: evolutionary relevance of crossfertility tests. J. Zool. 253: 25-32.
Gajardo, G. & Beardmore, J. A. 2001. Coadaptation: lessons from the brine
shrimp Artemia, “the aquatic Drosophila (Crustacea, Anostraca). Rev. Chil.
Hist. Nat., 74: 65-72
24
Gajardo, G., Beardmore, J. A. & Sorgeloos, P. 2001. Genomic relationships
between A. franciscana and A. persimilis, inferred from chromocentre
numbers. Heredity 82: 1-6.
Gajardo, G., Kappas, I., Abatzopoulos, Th. & Beardmore, J. A.. in press.
Evolution and speciation. In “Th. Abatzopoulos, J. A. Beardmore, J. S. Clegg
& P. Sorgeloos (eds) “Artemia: basic and applied biology”. Kluwer Academic
Publishers, Dordrecht.
25
INCO partner: 15
Partner name: IART: Institute for Artemia Research and Training,
Manonmaniam Sundaranar University, Rajakkamangalam, 629 502,
Kanyakumari District, Tamil Nadu, India
tel.: 91-4652-33680; fax: 91-4652-21457; email: petermar@md5.vsnl.net.in
Responsible scientist: Peter Marian
Author: Peter Marian
Title: Research on Artemia in the Institute for Artemia Research and Training
Abstract:
We have been working on Artemia in India since 1990. India has 1,78,848 ha
area of saltworks in the east and west coast, covering 8000 km length. A new
site in south India, called Thamaraikulam, was identified where only
parthenogenetic Artemia was identified and the characterization of the
Artemia parthenogenetica was undertaken. A detailed study on the salt and
Artemia production in the two districts of Tamil Nadu was undertaken during
the period 1995 to 2000. Artemia franciscana was cultured in the field in
collaboration with the private sector: one on the east coast (8°50'N 78°08'E)
and another on the west coast, North India (23°00’N 70°10'E). Cyst
production was 4.2 kg/ha/month. As there is a restriction on the use of urea to
produce microalgae in the saltworks by the salt department, Govt. of India, a
manure development research programme is going on in which urea is being
replaced by adding other nitrogenous nutrients, supplying manure as well as
choosing nitrogen-fixing microalgae.
The universal use of rice bran food in Artemia culture is being supplemented
and replaced with other ingredients such as grass powder and seaweed
(macroalgae) powder. Bio-medicinal products supplementation was achieved
in trace quantity with profitable production. Further improvements on the use
of marine yeasts (raised using the locally available cheap nutrients) were also
studied. The marine yeast produced using the sugar industry wastes was
found to be highly suitable for ensuring the growth of Artemia.
Artemia franciscana cysts were inoculated in the saltworks of the
Kanyakumari district to study the viability and growth in the saltworks. The
phenotypic and genotypic characteristics of the Artemia are being
studied. Biotechnical application of the salt purification using Artemia and
halophilic bacteria is also underway. Cyst production was induced using
some chemicals and bio-medicinal products. The cysts produced were of
good quality. Also the cyst induced protein(s) of local strains are studied in
collaboration with the Biophysics and Marine Biotechnology Departments.
So far 11 students have completed their Ph.D on various aspects of Artemia
research and 6 students are pursuing their Ph.D on Artemia in IART. 27
Students have completed their M.Sc degree on Artemia research projects in
IART. IART also collaborates with the other salt and other fisheries and
MPEDA departments. Students/scientists from research and education
departments from various parts of India are being helped in their research and
education programmes in connection with the Artemia research. We wish to
develop it as the centre for SAARC countries/Indian subcontinent.
26
Abstracts - Guests
Guest speaker: Elena Boyko
Affiliation: Ministry of Agriculture and Food Stuffs of the Russian
Federation, Association “State – cooperative union of fish industry”,
Federal state unitary enterprise, The Siberian Science-Research and
Project-Construction Institute of Fishery (“SIBRYBNIIPROJECT”),
625023, Odesskaya st., 33, Tyumen, Russia.
tel. 3452 22-41-16, 22-54-62; fax: (3452) 22-61-96; email: lotsman@sibtel.ru
Authors: Elena Boyko, Peter Bossier
Title: Authentication of Artemia from Russia
Abstract:
In Russia Artemia is spread in western Siberia and the Far East. The salinity
of Russian salt lakes varies from 10 till 340 ppt. Dates of many years show
that biomass of Artemia in lakes of western Siberia is 1.43-310.0 g/m3.
Scientists of SibrybNIIproject investigated 29 lakes, with a total surface area
of 281.4 km2. Artemia is found in 28 lakes.
Our research focuses on studying the genetic differentiation of Russian
Artemia populations. Ten strains have been used in the experimental DNA
analysis. The PCR reaction is based on the investigation of 1564 bp
mitochondrial ribosomal DNA fragments. The first results with use of RFLP
analysis show that this fragment is monomorphic between the tested strains.
The UPGMA dendrogram illustrates how these Russian strains relate to other
strains in the database.
27
Guest speaker: Luc Brendonck
Affiliation: Laboratory Aquatic Ecology, KULeuven, Ch. De Bériotstraat
32, 3000 Leuven, Belgium
tel. : 32-16-323714 ; fax : 32-16-324575 ;
email :Luc.Brendonck@bio.kuleuven.ac.be
Author: Luc Brendonck
Title: Adaptations to life in hyper-oligotrophic ephemeral pools
Abstract:
Branchipodopsis species are particularly dominant in small short-lived and
clear rock pools, to the vagaries of which they are extremely well adapted.
Common features of pools on granite outcrops are the clear state of the water
and the very low conductivity values (generally below 50 µS cm -1, often less
than 10 µS cm-1). These shallow (usually less than 50 cm) water bodies with
limited buffering capacity show major fluctuations in pH values (often between
about pH 4 and 11), depending on the time of the day and the stage in the
hydrocycle. Rock pools also closely follow ambient air temperatures with
resulting fluctuations of values between 10 and 40°C. Depending on local
climatic conditions, small rock pools are usually short-lived and have several
wet/dry cycles during one rainy season. Branchipodopsis species are the
record-holders in this race against time with maturation usually occurring
within the first week after inundation. Broods of resting eggs are generally
small (less than 80 eggs) but are produced almost daily. Often (in 30% in
south-eastern Botswana), erratic rainfall does not allow sufficient time for
successful maturation and reproduction. As a safeguarding mechanism in B.
wolfi from southeastern Botswana, only part of the eggs hatches at each
occasion, depending on environmental conditions and conform the chances
for successful recruitment. The vast egg banks in most pools illustrate
successful adaptation to this marginal habitat. While short-range dispersal of
floating eggs by overflows is common, long-range dispersal (e.g. by wind)
seems to be rare and restricted to shallow pools with little vegetation. Limited
effective dispersal may explain the high number of endemic species (e.g. 4 in
the Drakensberg region) and the large morphological variation in some
widespread species (e.g. B. wolfi).
Relevant publications:
Brendonck, L. & B. J. Riddoch. 2000a. Egg bank dynamics in anostracan desert
rock pool populations (Crustacea: Branchiopoda). Arch. Hydrobiol. 148: 71-84.
Brendonck, L. & B.J. Riddoch. 2000b. Dispersal in the desert rock pool
anostracan Branchipodopsis wolfi (Crustacea: Branchiopoda). Crust. Iss. 12:
109-118.
Brendonck, L. & W.D. Williams. 2000. Biodiversity in wetlands of dry regions
(drylands). In: Gopal, B., W.J. Junk & J.A. Davis (eds.), Biodiversity in wetlands:
assessment, function and conservation, vol. 1, pp. 181-194. Backhuys
Publishers, Leiden, The Netherlands.
Brendonck, L., L. De Meester & B.J. Riddoch. 2000. Regional structuring of
genetic variation in short-lived rock pool populations of Branchipodopsis wolfi
(Crustacea: Anostraca). Oecologia 123: 506-515.
28
Brendonck, L., E. Michels, L. De Meester & B.J. Riddoch. in press. Temporary
pools are not “enemy-free”. Hydrobiologia.
29
Guest speaker: Godelieve Criel
Affiliation: Department of Human Anatomy, University of Ghent,
Belgium,
tel: 32-9-2649230, fax: 32-9-2649498, email:godelieve.criel@rug.ac.be
Author : Godelieve Criel
Title: Morphological tools to help distinguishing Artemia species and
populations
Abstract:
Since the foundation of taxonomy by Linnaeus morphological aspects have
been used to distinguish species. As regards Artemia, Schmankewitsch
(1875) was the first to warn that environmental changes may induce
morphological changes. So a general agreement developed that also other
approaches are needed to characterise a species. Nonetheless morphological
examination is useful because it gives at least a hint. Since the work of
George Triantaphyllidis (1997) the morphology of penes has been used to
help species identification in the bisexual species of Artemia. Not only does a
quick check of penis morphology allow us to easily recognize a contamination
of A. salina with A. franciscana in Old World locations, it also permits to
indicate sites where interesting populations can be found. From all
populations studied thus far, only A. salina is devoid of penis adornments: A.
franciscana, persimilis, sinica and tibetiana all have a protuberance at the
penile base. The two Lybian populations examined show small spines instead
of a protuberance decorating the testis base, which invites further study. Luc
Brendonck used the ornamentations of the eversible penis in the diagnosis of
several Branchipodidae. Laura Torrentera and Denton Belk compared A.
franciscana and A. persimilis in this respect (paper in press in Hydrobiologia):
unlike Wolfe (1971), who described three to five spines, they found no spines
at the base of the eversible penis in A. franciscana and several spine-like
outgrowths in A. persimilis, and this while the study of the protuberances on
the proximal part of the penis does not allow irrefutable discrimination
between them. Cross breeding tests of A. franciscana females with A. sinica
males showed to be letal for most females, and no female ever survived a
second meeting. A comparison of the smooth everted penis found in a
population of Basrah with the barbed one found in male A. sinica suggests the
reason.
L. Brendonck (1995): An updated diagnosis of the branchipodid genera
(Branchiopoda: Anostraca: Branchipodidae) with reflexions on the genus
concept by Dubois (1988) and the importance of genital morphology in
anostracan taxonomy. Arch. Hydrobiol./Suppl. 107
G.V.Triantaphyllidis, G.R.J.Criel, T.J.Abatzopoulos, K.M.Thomas, J. Peleman,
J.A.Beardmore, P.Sorgeloos (1997): International study on Artemia. LVII.
Morphological and molecular characters suggest conspecificity of all bisexual
European and North African Artemia populations.
Marine Biology 129 477-487
30
Guest speaker: Jim Clegg
Affiliation: Bodega Marine Laboratory, and Section of Molecular and
Cellular Biology, University of California, Davis. Bodega Bay, CA 94923
USA
tel.: 1-707 875-2010; fax: 1-707 875-2009; email: jsclegg@ucdavis.edu
Authors: Clegg, J.S., Willsie, J.K., Frankenberg, M.M., Viner, R.I. and
Jackson, S.A.
Title: The diversity of Artemia habitats is reflected in the biochemical
repertoire of this remarkable organism
Abstract:
Artemia can be considered as an extremophile of the animal kingdom in the
sense that it lives, and often thrives, in environments in which few (or no)
other animals can exist. Thus, the encysted gastrula embryo (cyst) is arguably
the most resistant of all animal life history stages to extremes of
environmental stress, ranging from severe desiccation to thermal stress and
prolonged anoxia, while the motile stages appear to be the best
osmoregulators in the entire animal kingdom. These capabilities, to be
summarized in the presentation, provide a clear and fundamental basis for the
ecological success of Artemia, world-wide. That success depends on a variety
of biochemical, biophysical and physiological adaptations acquired during its
long evolutionary history.
These include the activities of various
osmoregulatory structures of the larvae and adults and, in the cysts, the
accumulation of large concentrations of the disaccharide trehalose and the
polyol glycerol, the synthesis of an extremely abundant and diverse guaninecontaining nucleotide pool, and the presence of massive amounts of a heat
shock (stress) protein called p26 that acts as an effective molecular
chaperone. How these function in survival, and how they are related to the
natural setting of Artemia will be considered.
Some of what we have learned about the molecular and cellular biology of
Artemia encysted embryos is currently being applied to the stabilization of
mammalian cells in culture. This exploratory research, just under way, is a
cooperative effort between our laboratory and that of Dr. Tom MacRae,
Dalhousie University, and is another example of the value of this model
system.
(JSC acknowledges support from United States NSF grant MCB-98 07762)
Relevant publications:
Liang, P., R. Amons, T.H. MacRae & J.S. Clegg. 1997. Purification, structure
and molecular chaperone activity in vitro of Artemia p26, a small heat shock
-crystallin protein. Eur. J. Biochem. 243:225-232.
Clegg, J.S., S.A. Jackson, N.V. Hoa & P. Sorgeloos. 2000. Thermal
resistance, developmental rate and heat shock proteins in Artemia
franciscana from San Francisco Bay and southern Vietnam. J. Exp. Mar. Biol.
Ecol. 252:85-96.
Willsie, J.W. & J.S. Clegg. 2001. Nuclear p26, a small heat shock -crystallin
protein, and its relationship to stress resistance in Artemia embryos. J. Exp.
Biol. 204:2339-2350.
31
Clegg, J.S., N.V. Hoa & P. Sorgeloos. 2001/02, in press. Thermal tolerance
and heat shock proteins in encysted embryos of Artemia from widely different
thermal habitats. Hydrobiologia (symposium chapter)
Willsie, J.K. & J.S. Clegg. 2002. The small heat shock protein p26 associates
with nuclear lamins and hsp70 in nuclei and nuclear matrix fractions from
stressed cells. J. Cell. Biochem. 84:601-614.
Clegg, J.S. and C.N.A. Trotman. 2002, in press. Physiological and
biochemical aspects of Artemia ecology. In: Artemia: Basic and Applied
Biology. Th. J. Abatzopoulos, J.A. Beardmore, J.S. Clegg and P. Sorgeloos
(Eds), Kluwer Academic Publishers, Dordrecht, The Netherlands.
32
Guest speaker: Luc De Meester
Affiliation: Laboratory Aquatic Ecology, KULeuven, Ch. De Bériotstraat
32, 3000 Leuven, Belgium
tel. :
32-16-323708 ;
fax :
32-16-324575 ;
email :
Luc.DeMeester@bio.kuleuven.ac.be
Author: Luc De Meester
Title: Dispersal, habitat
zooplankton populations
size
and
genetic
differentiation
among
Abstract:
We will provide a short introduction to zooplankton population genetics, with
emphasis on the influence of habitat size on genetic diversity within and
genetic differentiation among populations. We will emphasize the
observations that high dispersal capacities do not always translate in high
levels of gene flow, and that founder events are very important in zooplankton
population genetics.
Relevant publications:
De Meester, L., 1996. Local genetic differentiation and adaptation in freshwater
zooplankton populations: patterns and processes. Ecoscience 3: 385-399.
Naihong, X., E. Audenaert, J. Vanoverbeke, L. Brendonck, P. Sorgeloos & L. De
Meester, 2000. Low among-populational genetic differentiation in Chinese
bisexual Artemia populations. Heredity 84: 238-243.
Cousyn, C., L. De Meester, J.K. Colbourne, L. Brendonck, D. Verschuren & F.
Volckaert, 2001. Rapid local adaptation of zooplankton behavior to changes in
predation pressure in absence of neutral genetic changes. Proceedings of the
National Academy of Sciences USA 98: 6256-6260.
Michels, E., K. Cottenie, L. Neys, K. De Gelas, P. Coppin & L. De Meester,
2001. Modelling the effective geographic distance between zooplankton
populations in a set of interconnected ponds : a validation with genetic data.
Molecular Ecology 10: 1929-1938.
De Meester, L., A. Gómez, B. Okamura & K. Schwenk, in press. Dispersal,
monopolisation and (the lack of) gene flow in continental aquatic organisms.
Acta Oecologia.
33
Guest Speaker: Tom MacRae
Affiliation: Department of Biology, Dalhousie University, Halifax, N.S.,
B3H 4J1 Canada
tel.: 902-494-6525; fax: 902-494-3736; email: tmacrae@is.dal.ca
Authors: MacRae, T. H., Chen, T., Gupta, J., Sun, Y., Qiu, Z., Oulton, M. and
Malaviarachchi, K.
Title : Microtubule proteins and chaperones: molecular technologies
applied to study of the brine shrimp, Artemia franciscana
Abstract:
The brine shrimp, Artemia franciscana, has been used as an experimental
animal in our laboratory for analysis of the cytoskeleton and, more recently,
for observation of molecular chaperones during embryo development. In the
former case, tubulin and microtubule-associated proteins were the focus,
including isolation of their genes, biochemical characterization of the proteins,
and subcellular localization by immunofluorescent staining of larvae followed
by confocal microscopy. Work on chaperones involved similar approaches,
with emphasis on a small heat shock/α-crystallin protein, termed p26, during
encystment and diapause of oviparously developing embryos. The
experiments have medical implications because p26 shares sequence
homology with lens crystallins, disruption of which leads to cataract. To
continue these projects we are exploiting an expanding range of molecular
and analytical tools, common to the study of many organisms, but less so for
crustaceans. Both cDNA and genomic libraries were generated for Artemia,
with the cDNA library employed for construction of an expressed sequence
tag (EST) library from which many interesting full length clones have been
recovered. Artemia cDNA was expressed in prokaryotic and eukaryotic cells in
order to determine the structural/functional properties of p26. The cDNA was
modified by site-directed and deletion mutagenesis allowing appraisal of
discrete p26 domains and amino acid residues. Additionally, transgenic
Drosophila are being produced and will be used to inspect p26 in different
developmental stages of the organism, thereby permitting direct comparison
to results obtained by investigation of Artemia. Proteins that interact with p26
were captured by affinity chromatography, resolved in two dimensional gels
and sequenced by mass spectrometry in a mini-proteomics approach, a
procedure we will employ for identification of proteins that bind microtubules.
Clearly, the number of techniques that can be utilized to examine Artemia, in
concert with its unusual life history, make it an extremely interesting organism
for cell/molecular research.
THM acknowledges the financial support of the Natural Sciences and
Engineering Research Council of Canada and the Nova Scotia Health
Research Foundation.
Relevant publications:
Crack, J.A., Mansour, M., Sun, Y. & MacRae, T.H. 2002. Functional Analysis
of a Small Heat Shock/α-Crystallin Protein from Artemia franciscana:
Oligomerization and Thermotolerance. Eur. J. Biochem. 269: (in press).
34
Day, R., Criel, G.R.J., Walling, M.A. & MacRae, T.H. 2000. Posttranslationally
Modified Tubulins and Microtubule Organization in Hemocytes of the Brine
Shrimp, Artemia franciscana. J. Morph. 244: 153-166.
Liang, P & MacRae, T.H. 1999. The Synthesis of a Small Heat Shock/αCrystallin Protein in Artemia and Its Relationship to Stress Tolerance during
Development. Develop. Biol. 207: 445-456.
Walling, M.A., Criel, G.R.J. & MacRae, T.H. 1998. Characterization of γTubulin in Artemia: Isoform Composition and Spatial Distribution in Polarized
Cells of the Larval Epidermis. Cell Motil. Cytoskel. 40: 331-341.
Liang, P., Amons, R., Clegg, J.S. & MacRae, T.H. 1997. Molecular Structure
of a Small Heat Shock/α-Crystallin Protein in Encysted Artemia Embryos.
J.Biol. Chem. 272: 19051-19058
35
Guest speaker: Brad Marden
Affiliation: Research and Development, Utah Strategic Alliance, 5859
North Cottonwood Canyon Rd., Mountain Green, Utah, 84050, USA
work phone: 801-876-2400; mobile phone: 801-554-5163 ; fax: 801-554-7100;
email: mardenb@attglobal.net
Author: Brad Marden
Title: Recovery of the Great Salt Lake, USA, brine shrimp (Artemia
franciscana) population
Abstract:
Brine shrimp biomass and cysts found in the Great Salt Lake, Utah, USA have
been exploited as a commercial resource since the 1950s, however the brine
shrimp industry has only been intensively managed since the 1990’s.
Research-based management practices were implemented in 1996 to ensure
the long-term sustainability of the resource as well as to protect the integrity of
the Great Salt Lake ecosystem. During the period from 1996 to 1999 the
Artemia population declined substantially resulting in changes in the manner
in which the brine shrimp industry was regulated and in the enactment of other
mitigating measures to remedy the declining Artemia population. Over the
past two years the population has rebounded and the commercial industry has
harvested record amounts of Artemia cysts. Population dynamics, weather
conditions, phytoplankton, industry cooperation and management all
contributed to the recovery of the Great Salt Lake Artemia population. I
discuss in detail the population dynamics and the future potential for the Great
Salt Lake Artemia population.
36
List of participants
Abatzopoulos Theodore, Aristotle University of Thessaloniki, Department of
Genetics, Development & Molecular Biology, Faculty of Sciences, School of
Biology, Thessaloniki, Greece, tel: 30-31-998301, fax: 30-31-998256, email:
abtzop@bio.auth.gr
Agh Naser, Artemia and Aquatic Animals Research Center, Urmia University,
165 Shahid Beheshty Av. 57 153 Urmia, Iran, 98-441-440295, fax: 98-441440295, email: agh1960@hotmail.com
Amat Francisco, Instituto de Acuicultura de Torre de la Sal (Consejo
Superior de Investigaciones Científicas de España), Ribera de Cabanes
(Castellón), Spain, tel : 34-964-319500, fax : 34-964-319509, email :
amat@iats.csic.es
Baxevanis Thanos, Aristotle University of Thessaloniki, Department of
Genetics, Development & Molecular Biology, Faculty of Sciences, School of
Biology, Thessaloniki, Greece, tel: 30-31-998301, fax: 30-31-998256
Beardmore John, University College of SwanseaSchool of Biological
Sciences, University of Wales, Singleton Park, Swansea SA2 8PP, UK
tel.+44-792-205678, fax: +44-792-29561,
email: j.a.beardmore@swansea.ac.uk
Bossier Peter, Agricultural Research Center-Ghent/Department of Sea
Fisheries, Ankerstraat 1, 8400 Ostend, Belgium, tel: 32- 59 –342266 , fax: 3259-330629, email:pbossier@yucom.be
Boyko Elena, Tyumen State Agricultural Academy - The Siberian ScienceResearch and Project-Construction Institute of Fishery Tyumen, Russia tel:
3452-22-41-16, fax: 3452-22-61-96, email: lotsman@sibtel.ru
Brendonck Luc, Laboratory Aquatic Ecology, KULeuven, Ch. De Bériotstraat
32, 3000 Leuven, Belgium, tel. : 32-16-323714 ; fax : 32-16-324575 ; email :
Luc.Brendonck@bio.kuleuven.ac.be
Camara Marcos, Departamento de Oceanografia e Limnologia, Universidade
Federal do Rio Grande do Norte, Campus Universitario, BR 101, 59072-970
Natal,
Brazil,
tel.:
55-84-2154433;
fax:
55-84-6421815;email:
mrcamara@ufrnet.br
Castro Mejia Jorge, Universidad Autónoma Metropolitana-Xochimilco, Depto.
El Hombre y su Ambiente, Calz. Del Hueso No. 1100. Col. Villa Quietud,
Mexico 04960, D.F. Mexico, tel: 52-5-4837151, fax: 52-5-4837469, email:
cabt7515@cueyalt.uam.mx
Clegg Jim, Bodega Marine Laboratory, Section of Molecular and Cellular
Biology, University of California, Davis, Bodega Bay, USA, tel:1-707-8752010,
fax:1-707-8752009, email: jsclegg@ucdavis.edu
37
Criel Godelieve, Department of Anatomy, Embryology and Histology, Ghent
University,
Belgium,
tel:
32-9-2649230,
fax:
32-9-2649498,
email:godelieve.criel@rug.ac.be
De Meester Luc, Laboratory Aquatic Ecology, KULeuven, Ch. De Bériotstraat
32, 3000 Leuven, Belgium, tel. : 32-16-323708 ; fax : 32-16-324575 ; email :
Luc.DeMeester@bio.kuleuven.ac.be
Gajardo Gonzalo, Department of Basic Sciences/Laboratory of Genetics &
Aquaculture, Universidad de Los Lagos, P.O. Box 933, Avenida Fuchslocher
s/n, Osorno, Chile, tel. 56-64-205293; fax: 56-64-239517; email:
ggajardo@ulagos.cl
Hecht Tom, Department of Ichthyology and Fisheries Science, Rhodes
University, Grahamstown, South Africa, tel: 27-46-6038415, fax: 27-466224627, email: t.hecht@ru.ac.za
Kaiser Horst, Department of Ichthyology and Fisheries Science, Rhodes
University, Grahamstown, South Africa, tel: 27-46-6038415, fax: 27-466224627,email: h.Kaiser@ru.ac.za
MacRae Tom, Department of Biology, Dalhousie University, Halifax, N.S.,
Canada, tel: 1-902-494-6525, fax: 1-902-494-3736, email: tmacrae@is.dal.ca
Marden Brad, Research and Development, Utah Strategic Alliance, 5959
Northe Cottonwood Canyon rd. Mountain Green, Utah, 84050, USA, tel: 1801-876-2400, fax: 1-801-554-7100, email: mardenb@attglobal.net
Maryan Peter, Institute for Artemia Research and Training, Manonmaniam
Sundaranar University, Rajakkamangalam, 629 502, Kanyakumari District,
Tamil Nadu, India, tel.: 91-4652-33680; fax: 91-4652-21457; email:
petermar@md5.vsnl.net.in
Mura Graziella, Dipartamento di Biologia Animale é dell’Uomo, Laboratorio di
Zoologia Applicata, Università La Sapienza, Rome, Italy, tel : 39-6-49914769,
fax :39-6-4958259, email : graziella.mura@uniroma1.it
Nguyen Van Hoa, Institute of Science for Aquaculture, Can Tho University,
Can Tho, Vietnam, tel: 84-71-834307, fax: 84-71-830323, email:
nvhoa@stu.edu.vn
Romdhane Mohammed, Institut National Agronomique de Tunisie, Université
de Carthage, Tunis, Tunisia, tel : 216-1-287110, fax : 216-1-799391, email :
rhomdhane.medsalah@inat.agrinet.tn
Sorgeloos Patrick, Laboratory of Aquaculture & Artemia Reference Center,
Ghent University, Belgium, tel: 32-9-2643754, fax: 32-9-2644193, email:
Patrick.Sorgeloos@rug.ac.be
Van Stappen Gilbert, Laboratory of Aquaculture & Artemia Reference
Center, Ghent University, Belgium, tel: 32-9-2643754, fax: 32-9-2644193,
email: gilbert.vanstappen@rug.ac.be
38
Xin Naihong, Salt Research Institute, Yingkou Road 831, 300450 Tanggu,
Tianjin, PR China, tel: 86-22-25301094, fax: 86-22-25301094, email:
srisalt@public.trp.tj.cn
Wang Xiaomei, Agricultural Research Center-Ghent/Department of Sea
Fisheries, Ankerstraat 1, 8400 Ostend, Belgium, tel: 32- 59 –342266 , fax: 3259-330629, email:srisalt@public.trp.tj.cn
39