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:
1
Programme
Tuesday February 5: Oral presentations
8.30-8.45Patrick Sorgeloos: Welcome & introduction
8.45-9.05Gilbert Van Stappen, Belgium (INCO 1)
9.05-9.25Peter Bossier, Belgium (INCO 2)
Title: A database for Artemia Authentication
9.25-9.45Elena 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)
10.25-11.00break
11.00-11.20Graziella Mura, Italy (INCO 5)
11.20-11.40Godelieve 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
12.00-12.20Mohammed Romdhane, Tunisia (INCO 8)
Title: Status of the Artemia Biodiversity in Tunisian salt areas
12.20-13.30lunch pauze
13.30-13.50Tom MacRae, Canada (Guest)
Title: Microtubule proteins and chaperones: molecular technologies applied to study of the brine shrimp, Artemia franciscana
13.50-14.10Jim Clegg, USA (Guest)
Title: The diversity of Artemia habitats is reflected in the biochemical repertoire of this remarkable organism
14.10-14.30Horst Kaiser, South Africa (INCO 9)
Title: Characterisation of southern African Artemia populations, and the use of Artemia as a vector for probionts and hormones
14.30-14.50Xin Naihong, China (INCO 10)
14.50-15.10Naser Agh, Iran (INCO 11)
15.10-15.50break
15.50-16.10Brad Marden, Canada (Guest)
Title: Recovery of the Great Salt Lake, USA, brine shrimp (Artemia franciscana) population
16.10-16.30Jorge Castro Mejia, Mexico (INCO 12)
Title: Artemia research in the Universidad Autonoma Metropolitana-Xochimilco, Mexico
16.30-16.50Marcos Camara, Brazil (INCO 13)
Title: Artemia research at Universidade Federal do Rio Grande do Norte (UFRN), Natal, Brazil
16.50-17.10Gonzalo Gajardo, Chile (INCO 14)
Title: Artemia characterization, species and speciation
17.10-17.30Peter Maryan, India (INCO 15)
Title: Research on Artemia in the Institute for Artemia Research and Training
17.30-17.45Luc De Meester, Belgium (Guest)
Title:Dispersal, habitat size and genetic differentiation among zooplankton populations
17.45-18.00Luc 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
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:
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 theLaboratory 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.
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
INCO partner: 2
Partner name: CLO-DVZ: Agricultural Research Center-Ghent/Department of Sea Fisheries (CLO-DVZ), Ankerstraat 1, 8400 Oostende, Belgium
tel. 32-9-34 22 66; fax: 32-9-33 06 29; email:
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.
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:
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.
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)
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:
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 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): 329-340.
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.
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:
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.
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.
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:;
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.