A note on the brine shrimp Artemia in Tuz Lake (Turkey)

Population density, fecundity andreproduction period of Artemia sp. in TuzLake, Turkey, beside environmental factors, such as oxygen,temperature, salinity, pH and electrical conductivity werestudied during 1993 and 1994.Artemia sp. occurred at two stations at the coast.Overwintering cysts hatched in early May, when the population was composed of 90% metanauplii (93%at Y4 and 99% at Y5) and few adult females. Peak abundanceoccurred in late May, at 106to 114 ind.m^–3.     

Genetic variation in sympatric populations of diploid and polyploid brine shrimp (Artemia parthenogenetica)

We examined genetic variation in sympatric diploid and polyploid brine shrimp Artemia parthenogenetica from each of three populations (China, Italy and Spain). Italian and Spanish tetraploids are closely related (I=0.964). Diploids and tetraploids within each of the two European populations are also closely related (mean I=0.905). Most alleles found in diploids also exist in sympatric polyploids. In contrast, the asexual Artemia (2N, 4N and 5N) in our study share few alleles with their close sexual relative, A. tunisiana (mean I=0.002). These results, as well as the work of other authors, strongly suggest that at least the tetraploid Artemia in our study have an autopolyploid origin.Clonal diversity of polyploid Artemia can be very high at least in some population. Both diploids and polyploids had low clonal diversities in the populations dominated by polyploids and high clonal diversities in the population dominated by diploids.The most common genotypes of sympatric diploid and polyploid Artemia frequently differed. Some alleles occurred only in diploids, while others were restricted to polyploids. These results suggest that polyploidy in Artemia has led to genetic divergence from diploid progenitors, and that ploidy-level variation must also be considered in developing an understanding of spatial and temporal allozyme polymorphism in asexual populations.     

Characterization of polymorphic microsatellite markers in the brine shrimp Artemia (Branchiopoda, Anostraca)

The brine shrimp Artemia is a complex genus containing sexual species and parthenogenetic lineages. Artemia franciscana is native to America and its cysts (diapausing eggs) are used worldwide as a food source in aquaculture. As a consequence, this anostracan has become an invasive species in many hypersaline aquatic ecosystems of other continents. Parthenogenetic Artemia lineages occur only in the Old World. Ten and five microsatellite markers were developed to characterize two populations for A. franciscana and two populations for diploid parthenogenetic Artemia, respectively. For A. franciscana the number of alleles ranged from 11 to 58 per locus, while for parthenogens the number of alleles ranged from three to 10. The levels of heterozygosity in A. franciscana and in parthenogens ranged from 0.115 to 0.976 and from 0.000 to 0.971, respectively. These microsatellite loci showed a high population assignment power, which will be useful for future studies of population genetics and invasive processes in Artemia.     

Developmental and comparative aspects of brine shrimp tubulin.

Tubulin from embryos of the brine shrimp Artemia has been purified to apparent homogeneity by chromatography on phosphocellulose P11 and DEAE-cellulose, (NH4)2SO4 fractionation and assembly-disassembly of microtubules. Peptide mapping indicated that Artemia and bovine brain tubulin were very similar in spite of differences in the electrophoretic behaviour of tubulin from these two organisms. Isoelectric focusing and two-dimensional gel electrophoresis were used to resolve and identify several Artemia isotubulins . The isotubulin composition and the quantity of tubulin did not change during pre-emergence development of Artemia embryos. Formation of microtubules with tubulin purified from embryos at different stages of development did not require glycerol or microtubule-associated proteins and formation of structurally normal microtubules was actually hindered by glycerol and Mg2+. The characteristics of Artemia tubulin, in concert with the unusual life history of Artemia, suggest that this organism will be very useful for the study of tubulin gene expression and tubulin utilization during embryo development.     

Molecular phylogenetics and asexuality in the brine shrimp Artemia

Explaining cases of long-term persistence of parthenogenesis has proven an arduous task for evolutionary biologists. Interpreting sexual-asexual interactions though has recently advanced owing to methodological design, increased taxon sampling and choice of model organisms. We inferred the phylogeny of Artemia, a halophilic branchiopod genus of sexual and parthenogenetic forms with cosmopolitan distribution, marked geographic patterns and ecological partitioning. Joint analysis of newly derived ITS1 sequences and 16S RFLP markers from global isolates indicates significant interspecific divergence as well as pronounced diversity for parthenogens, matching that of sexual ancestors. Maximum parsimony, maximum likelihood, and Bayesian methods were largely congruent in reconstructing the phylogeny of the genus. Given the current sampling, at least four independent origins of parthenogenesis are deduced. Molecular clock calibrations based on biogeographic landmarks indicate that the lineage leading to A. persimilis diverged from the common ancestor of all Artemia species between 80 and 90 MYA at the time of separation of Africa from South America, whereas parthenogenesis first appeared at least 3 MYA. Common mitochondrial DNA haplotypes delineate A. urmiana and A. tibetiana as possible maternal parents of several clonal lineages. A novel topological placement of A. franciscana as a sister clade to all Asian Artemia and parthenogenetic forms is proposed and also supported by ITS1 length and other existing data.     

Purification and characterization of phenoloxidase from brine shrimp Artemia sinica

Phenoloxidase from Artemia sinica (AsPO) was purified by Superdex 200 gel-filtration and Q Sepharose fast flow ion-exchange chromatography, and its properties were characterized biochemically and enzymatically by using L-dihydroxyphenylalanine (L-DOPA) as the specific substrate. Results showed that AsPO was isolated as a monomeric protein of 125.5 kDa in molecular mass. The optimal pH value and temperature are 7.0 and 50°C, respectively, for its PO activity. The AsPO had an apparent K(m) value of 4.2 mM on L-DOPA, and 10.9 mM on catechol, respectively. Oxidase inhibitor on PO activity showed that the AsPO was extremely sensitive to ascorbic acid, sodium sulfite, and citric acid; and was very sensitive to cysteine, benzoic acid, and 1-phenyl-2-thiourea. Combined with its specific enzyme activity on L-DOPA and catechol, it can be concluded that AsPO is most probably a typical catechol-type O-diphenoloxidase. Its PO activity was also sensitive to metal ions and chelators, and 20 mM DETC-inhibited PO activity was obviously recovered by 15 mM Cu(2+), indicating that AsPO is most probably a copper-containing metalloenzyme. All these data about specific substrate, sensitivity to oxidase inhibitor metal ions and chelators indicate that the AsPO has the properties of a catechol-type copper-containing O-diphenoloxidase that functions as a vital humoral factor in host defense via melaninization as in other Crustaceans.     

Development of the brine shrimp Artemia is accelerated during spaceflight

Developmentally arrested brine shrimp cysts have been reactivated during orbital spaceflight on two different Space Shuttle missions (STS-50 and STS-54), and their subsequent development has been compared with that of simultaneously reactivated ground controls. Flight and control brine shrimp do not significantly differ with respect to hatching rates or larval morphology at the scanning and transmission EM levels. A small percentage of the flight larvae had defective nauplier eye development, but the observation was not statistically significant. However, in three different experiments on two different flights, involving a total of 232 larvae that developed in space, a highly significant difference in degree of flight to control development was found. By as early as 2.25 days after reactivation of development, spaceflight brine shrimp were accelerated, by a full instar, over ground control brine shrimp. Although developing more rapidly, flight shrimp grew as long as control shrimp at each developmental instar or stage.     

The patterning function of purines in the brine shrimp Artemia

In two Artemia wild-type sibling species originating from the Old and New Worlds, the processes underlying the construction of the naupliar body during embryonic development and the construction of the adult body during postembryonic development are disrupted by specific nutritional deficiencies and/or the administration of metabolic inhibitors. The species-specific phenotypic outcomes of these experimental disruptions on the construction of segments and the establishment of their identity, permit us to outline a model in which Hox genes would act as intermediary cogwheels fastened to a mechanism put in gear upstream by purine-mediated processes which would trigger downstream folic acid-mediated processes. The prevalent view that Hox genes can select for different developmental programmes, is challenged by this model whose relevancy is analysed in the context of our present knowledge on the master functions ascribed to Hox genes in developmental and evolutionary processes.     

Antennular sensilla of the brine shrimp, Artemia salina.

1. Scanning electron microscopy was used to characterize the external morphology of setae found on the antennules of adults and nauplii of the brine shrimp, Artemia salina (L.). The permeability of the antennular setae was studied by means of Slifer's crystal violet method. 2. Each antennule of an adult brine shrimp possessed a terminal cluster of sensory setae. Within a cluster there were two morphologically distinct kinds of sensilla, here designated type 1 and type 2. Three type 1 sensilla were observed on every antennule examined. The number of type 2 sensilla per antennule was usually four or five. 3. Type 1 sensilla of adults were 43 to 80 micrometer long and simple in external morphology. They were widest at the base, decreased in diameter gradually, and terminated as a finely tapered tip. No pores were resolved by scanning electron microscopy. 4. Type 2 sensilla of adults were shorter (shaft length, 12 to 23 micrometer) and displayed a single pore at the tip (average pore diameter, 0.4 micrometer). In thin section they were seen to possess a distinctive articular specialization of the cuticle at the base of the seta. 5. Dye penetration experiments indicated that type 2 sensilla were permeable to aqueous crystal violet, whereas type 1 sensilla were not. 6. The antennular setae of nauplii resembled type 1 sensilla in general shape, in being impermeable to crystal violet, and in lacking a terminal pore and basal articular specialization. Moreover, a total of three setae was normally present on each naupliar antennule, and the same number of type 1 sensilla was found on each adult antennule examined. If the three naupliar setae represent a developmental stage in the formation of three adult sensilla, available observations suggest that the larval setae are developmentally related to type 1, rather than to type 2 adult sensilla.     

ZW sex-chromosome evolution and contagious parthenogenesis in Artemia brine shrimp

Eurasian brine shrimp (genus Artemia) have closely related sexual and asexual lineages of parthenogenetic females, which produce rare males at low frequencies. Although they are known to have ZW chromosomes, these are not well characterized, and it is unclear whether they are shared across the clade. Furthermore, the underlying genetic architecture of the transmission of asexuality, which can occur when rare males mate with closely related sexual females, is not well understood. We produced a chromosome-level assembly for the sexual Eurasian species Artemia sinica and characterized in detail the pair of sex chromosomes of this species. We combined this new assembly with short-read genomic data for the sexual species Artemia sp. Kazakhstan and several asexual lineages of Artemia parthenogenetica, allowing us to perform an in-depth characterization of sex-chromosome evolution across the genus. We identified a small differentiated region of the ZW pair that is shared by all sexual and asexual lineages, supporting the shared ancestry of the sex chromosomes. We also inferred that recombination suppression has spread to larger sections of the chromosome independently in the American and Eurasian lineages. Finally, we took advantage of a rare male, which we backcrossed to sexual females, to explore the genetic basis of asexuality. Our results suggest that parthenogenesis is likely partly controlled by a locus on the Z chromosome, highlighting the interplay between sex determination and asexuality.     

Evidence for chemically shifted water in the brine shrimp, (Artemia): possible interpretations

We report in this paper evidence for the existence of an unusually large chemical shift for intracellular water proton in brine shrimp (Artemia). The observed chemical shift, when corrected for bulk susceptibility, is down field 0.11 ppm from the pure water signal. The possible origin of such large chemical shift for intracellular water in this system is discussed.     

Lipid composition of three geographical sources of brine shrimp nauplii (Artemia sp.)

• 1.1. Percentage of triacylglycerols (TG), free fatty acids (FFA) and phospholipids (PL) in the total lipids, the fatty acid composition of each of these lipid classes, and the percentage of cholesterol were determined by gas chromatography in three geographical sources (San Francisco Bay, SFB; Chinese, CH; Colombian, COL) of brine shrim (Artemia sp.) nauplii.
• 2.2. There were no significant differences among sources of brine shrimp in total lipids, TG or FFA with means for all sources of 17.8, 65.8 and 10.9%, respectively. Percentage of phospholipid was significantly higher in SFB and CH sources of brine shrimp, 25.1 and 26.5%, respectively, than in COL 18.3%.
• 3.3. Marked differences in percentages of 18:3 (n-3) (linolenic acid) and 20:5 (n-3) (eicosapentaenoic acid or EPA) were found among brine shrimp sources, and concentration of these two fatty acids were usually inversely related within sources. The CH source contained higher concentrations of EPA ( > 9.0%) than the COL and SFB sources (< 5.0%) in all three lipid classes analyzed. No 22:6 (n-3) (docosahexaenoic acid or DHA) was found in any brine shrimp source.
• 4.4. Fatty acid compositions of the TG and PL were similar and did not differ among sources of brine shrimp, while the FFA had a lower percentage of polyunsaturated fatty acids, but was similar among sources of brine shrimp.
• 5.5. Differences in n-3 fatty acid composition indicated a difference in nutritional quality among sources of brine shrimp for feeding larval fish.

     

Structure of extracellular hemoglobin from the brine shrimp Artemia salina.

1. Hemoglobin from the brine shrimp Artemia salina, purified by ultracentrifugation and preparative gel electrophoresis in non-denaturing medium, gave in sodium dodecyl sulfate-polyacrylamide gel electrophoresis a single band corresponding to a polypeptide chain with Mr 150,000. 2. Crosslinking by glutardialdehyde resulted in the appearance of a band corresponding to a molecular mass twice that of a polypeptide chain. 3. Limited trypsinolysis gave eight proteolytic bands corresponding to submultiples 8/9-1/9 of a polypeptide chain. 4. We conclude that a molecule of Artemia hemoglobin is composed of two single polypeptide chain subunits and that each subunit consists of nine structural units roughly equal in size.     

DNA reassociation kinetic analysis of the brine shrimp, Artemia salina

DNA reassociation kinetics have been partly elucidated for the brine shrimp $\text{Artemia salina}$, using calf thymus DNA as a standard. The $\text{Artemia}$ single-copy DNA sequences comprise 45% of the genome; sequences having a repetition frequency of about 2–90 are not detectable. The average repetition frequency of the intermediately redundant DNA component is about 5,000 copies. Reassociation kinetic data are consistent with a unit genome size of 1.5 pg.     

Phylogeography and local endemism of the native Mediterranean brine shrimp Artemia salina (Branchiopoda: Anostraca)

There has been a recent appreciation of the ecological impacts of zooplanktonic species invasions. The North American brine shrimp Artemia franciscana is one such alien invader in hyper-saline water ecosystems at a global scale. It has been shown to outcompete native Artemia species, leading to their local extinction. We used partial sequences of the mitochondrial Cytochrome c Oxidase Subunit 1 (COI or cox1) gene to investigate the genetic diversity and phylogeography of A. salina, an extreme halophilic sexual brine shrimp, over its known distribution range (Mediterranean Basin and South Africa) and to assess the extent of local endemism, the degree of population structure and the potential impact of traditional human saltpan management on this species. We also examined the phylogenetic relationships in the genus Artemia using COI sequences. Our results show extensive regional endemism and indicate an early Pleistocene expansion of A. salina in the Mediterranean Basin. Subsequent population isolation in a mosaic of Pleistocene refugia is suggested, with two or three refugia located in the Iberian Peninsula. Two instances of long-distance colonization were also observed. Surprisingly, given its strong phylogeographical structure, A. salina showed a signature of correlation between geographical and genetic distance. Owing to strong 'priority effects', extensive population differentiation is retained, despite dispersal via migrant birds and human management of saltpans. The foreseeable expansion of A. franciscana is likely to be followed by substantial loss of genetic diversity in Mediterranean A. salina. Large genetic divergences between Mediterranean and South African A. salina suggest that the latter deserves species status.