LABORATORY OF AQUACULTURE & ARTEMIA REFERENCE …



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

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)

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 Metropolitana-Xochimilco, 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

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.

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: 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.

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.

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: 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 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: 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.

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: nvhoa@ctu.edu.vn; nvhoa.ct@bdvn.

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; 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.

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.

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 104 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

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.

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.

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.

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: 19-23.

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@

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.

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.

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 Metropolitana-Xochimilco, 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.

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.

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.

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: 117-123.

Gajardo, G., Parraguez, P. & Beardmore J.A. & Sorgeloos, P. 2001. Reproduction in the brine shrimp Artemia: evolutionary relevance of cross-fertility 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

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.

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..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. 

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.

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.

Brendonck, L., E. Michels, L. De Meester & B.J. Riddoch. in press. Temporary pools are not “enemy-free”. Hydrobiologia.

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

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 guanine-containing 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.

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.

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 size and genetic differentiation among zooplankton populations

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.

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).

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/α-Crystallin 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 γ-Tubulin 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

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@

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.

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: 98-441-440295, email: agh1960@

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: 32-59-330629, email:pbossier@yucom.be

Boyko Elena, Tyumen State Agricultural Academy - The Siberian Science-Research 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

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: 27-46-6224627, email: t.hecht@ru.ac.za

Kaiser Horst, Department of Ichthyology and Fisheries Science, Rhodes University, Grahamstown, South Africa, tel: 27-46-6038415, fax: 27-46-6224627,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: 1-801-876-2400, fax: 1-801-554-7100, email: mardenb@

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..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

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.

Wang Xiaomei, Agricultural Research Center-Ghent/Department of Sea Fisheries, Ankerstraat 1, 8400 Ostend, Belgium, tel: 32- 59 –342266 , fax: 32-59-330629, email:srisalt@public.trp.

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