Heterochromatin in the genus Artemia

A bisexual species of the genus Artemia (Crustacea, Phyllopoda), Artemia franciscana Barigozzi of San Francisco Bay and a parthenogenetic population of Artemia sp. of Tsing-Tao (China), both with 42 chromosomes, were compared with respect to the microscopic structure of the interphase larval nucleus, the microscopical structure of the prophase chromosomes and the DNA structure. — Artemia franciscana exhibits several chromocenters in the resting nucleus, heterochromatic blocks located at the end of the prophase chromosomes, and a large amount of repetitive DNA (Alu I 110-bp fragments). The other Artemia sp. lacks chromocenters, heterochromatic blocks in the chromosomes, and the Alu I DNA. The two populations thus differ by a remarkable amount of repetitive DNA.The authors dedicate this paper to Professor Hans Bauer, on the occasion of his 80th birthday     

Highly repetitive DNA sequence in parthenogeneticArtemia

Summary The study of the structural organization of the eukaryotic genome is one of the most important tools for disclosing the evolutionary relationships between species.Artemia (Crustacea, Phyllopoda) offers a very interesting model for speciation studies. The genus, distributed all over the world, comprises both bisexual sibling species and parthenogenetic populations, exhibiting different chromosome numbers (diploidy, polyploidy, and heteroploidy).Digestion of genomic DNA of the parthenogeneticArtemia sp. from Tsing-Tao (China) with the restriction enzymes Eco RI and Alu I reveals that a highly repetitive sequence of 133 bp is present. The Eco RI fragment has been cloned and characterized by genomic organization. The distribution of the Eco RI family of repeats was also studied in several bisexual and parthenogeneticArtemia populations and compared with an Alu I repetitive fragment previously identified inArtemia franciscana.     

Life history,resting egg formation,and hatching may explain the temporal-geographical distribution ofArtemia strains in the Mediterranean basin

Life history traits are presented for the sexual species (Artemia tunisiana) and for parthenogenetic diploid and tetraploid strainsA. parthenogenetica reared at 15 °, 24 ° and 29.5 °C.In laboratory cultures, we present evidence that the seasonal appearance of the sexualArtemia tunisiana (the dominant winter-spring population), and of two parthenogenetic populations ofArtemia (the dominant spring-summer populations) in certain Spanish saltworks is controlled by temperature through its effect on reproductive and survival traits. Minimum and maximum reproductive output and survival for the sexual and parthenogenetic populations, respectively, occurred at a typical temperature (24 °C) of the late-spring season when the sexual population is replaced by parthenogenetic forms. Furthermore, the high production and hatchability of cysts from the sexual population at low temperatures (15 °C), and of the parthenogenetic populations at middle temperature (24 °C), indicate the role of dormancy as an adaptation regulating seasonal occurrence.     

International study on Artemia XLIX. Salinity effects on survival,maturity, growth,biometrics, reproductive and lifespan characteristics of a bisexual and a parthenogenetic population of Artemia

Two Artemia populations, a bisexual from San Francisco Bay (California, USA) and a parthenogenetic from Tanggu area (Tianjin province, People's Republic of China) are assayed for their tolerance and fitness in various salinity levels. This study was carried out under laboratory conditions where salinity effects upon special characteristics of the two Artemia populations, such as survival, growth rate, maturation, morphology, fecundity and life duration, were recorded. This evaluation revealed that the two populations examined exhibits significant differences in their response against elevated salinity levels. Furthermore, specific biometric parameters can be a useful tool for the discrimination and/or determination of their distribution in a mixed population, which, in fact, is the case in Tanggu salt works. Artemia franciscana seems to be a more effective colonizer at higher salinities. The data presented in this study may generate useful suggestions for proper management of the solar saltworks at Tanggu although further experimentation is needed.International interdisciplinary study on Artemia populations coordinated by the Laboratory of Aquaculture & Artemia Reference Center, University of Ghent, Belgium.     

Estimating genetic and environmental components of variance using sexual and clonal Artemia

Reproductive and life span traits were measured for two obligately parthenogenetic (Artemia parthenogenetica) and three sexual (two A. franciscana and one A. sinica) brine shrimp populations. For each population, clonal lineages or single mating pairs were followed through one life cycle. The relative contributions of environmental and genetic components to total phenotypic variation for 10 life-history traits in response to environmental stress (0, 10, 25 ppb Cu) were estimated. Within treatment variation (CV_W) was 39% higher for sexual populations than parthenogenetic populations, with significant (p<0.05) differences in total number of offspring and number of nauplii. CV_A (the change in variance due to rearing in different environments), when averaged for all traits and all populations, increased variability by 9.9%. CV_A was 44.2% higher for sexual than parthenogenetic populations, with significant differences in number of broods, total number of offspring, and number of nauplii. The average genetic component of variation for the 10 traits was 23.44%, ranging from 5.26% for number of cysts to 44.87% for number of nauplii. For all traits, the environmental component of variance is greater than the genetic component measured, but every trait has a genetic component, which can potentially be acted upon by selection.     

Speciation in the Artemia genus: Mitochondrial DNA analysis of bisexual and parthenogenetic brine shrimps

From the cloned mitochondrial DNAs (mtDNAs) isolated from two bisexual species, one Mediterranean, Artemia salina, and one American, Artemia franciscana, and two parthenogenetic (diploid and tetraploid) strains of Artemia parthenogenetica collected in Spain, physical maps have been constructed and compared. They are extremely different among themselves, much more than the differences between Drosophila melanogaster and D. yakuba and in the same range of different mammalian species such as mouse/rat or man/cow. The nucleotide sequences of two regions of mtDNA encoding parts of the cytochrome c oxidase subunit I (COI) and cytochrome b (Cytb) genes have been determined in the two bisexual species and the two parthenogenetic strains. Comparisons of these sequences have revealed a high degree of divergence at the nucleotide level, averaging more than 15%, in agreement with the differences found in the physical maps. The majority of the nucleotide changes are silent and there is a strong bias toward transitions, with the CT substitutions being highly predominant. The evolutionary distance between the two Artemia parthenogenetica is high and there is no clear relationship with any of the bisexual species, including the one present nowadays in Spain. Using a combination of molecular (mtDNA) and morphological markers it is possible to conclude that all of these Artemia isolates should be actually considered as belonging to different species, even the two Artemia parthenogenetica diploidica and tetraploidica.On sabbatical leave from Departamento de Bioquímica, Facultad de Veterinaria, Universidad Complutense de Madridearly Italian artemiologists to designate the Medi-Beatriz Batuecas died in an accident during the Christmas holy days of 1988 after she had initiated this workCorrespondence to: R. Garesse     

Preliminary genetic data on some Caribbean Artemia franciscana strains based on RAPD's

A total of fourteen Artemia samples from Colombia, Venezuela, Curaçao (Netherlands Antilles), Puerto Rico, and reference samples from U.S.A. (San Francisco Bay, SFB) belonging to the superspecies Artemia franciscana, and Argentina (A. persimilis), were analysed with the RAPD technique in order to demonstrate genetic dissimilarities. Pearson's correlation coefficients between the DNA banding patterns were calculated. They served as input values for the construction of UPGMA dendrograms. The results indicate that, within the collection of Colombian, Venezuelan and the two Netherlands Antilles Artemiacyst samples examined, two different groups seem to exist. Geographically, the mountainous area of Sierra Nevada de Santa Marta separates these two groups (lower Caribbean to the South and middle Caribbean to the North). Although the Caribbean, North and South American populations belong to A. franciscana, genetic discontinuities are to be expected due to habitat differences and geographic isolation. The Sierra Nevada (with an altitude of about 5800 m) emerges as the barrier very likely to explain the observed RAPD differences. Little genetic variability was present in the Colombian samples from Manaure that were collected almost every ten years, nor in the samples from Galerazamba collected almost two decades apart, although these samples were more likely subjected to different prevailing environmental conditions. The SFB population did not show a very close relation with all Caribbean populations analyzed, including the Puerto Rican. All A. franciscana populations analyzed were divergent from A. persimilis(Argentina).     

Characterisation of high-altitude Artemia populations from the Qinghai-Tibet Plateau,PR China

The brine shrimp Artemia was discovered in a number of saline lakes on the Qinghai-Tibet Plateau, widely diverging in chemical composition. Several lakes were athalassohaline, with relatively high amounts of trace elements. Common environmental factors are their high altitude (exceeding 4500 m) and the low average annual temperatures. A number of Artemia populations in this area were analysed to assess their preference for low temperatures and an athalassohaline medium. Furthermore, their characteristics were compared with Artemia tibetiana, the species recently described for one lake in this area. All samples contained a variable mixture of parthenogenetic and bisexual individuals. A cross-breeding test of the sample from Jingyu Lake showed cross-fertility both with A. tibetiana and A. sinica. All populations showed similarities to A. tibetiana: a large cyst diameter and naupliar length, high HUFA content and a high tolerance to low temperatures, as compared to the control A. franciscana samples. These can thus be considered as recurrent characteristics of the populations from the high-altitude low-temperature environment on the Qinghai-Tibet Plateau, although further research is needed to identify their exact species status.     

Morphometric and Preliminary Genetic Characteristics of Artemia Populations from Iran

Six Artemia populations, one bisexual and five parthenogenetic, from different parts of Iran were compared using morphometric and genetic characteristics. The discriminant analysis based on 19 morphometric variables showed that there are significant differences between the studied populations based on their morphological characteristics, where 85.9% of original grouped cases were correctly classified. The bisexual Artemia urmiana however exhibited a 100% separation from the parthenogenetic populations. However, a 1500 bp mitochondrial rDNA fragment showed similar RFLP patterns for all Iranian populations confirming earlier reports of a close genetic relationship between A. urmiana and parthenogenetic Artemia. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Observations on future sea level changes in the Venice lagoon

The identification of the environmental conditions inducing different ecophysiological responses in the different strains and populations of the brine shrimp Artemia should improve the understanding of their biogeographic distribution. Nauplii from two Argentinean brine shrimp populations, Artemia persimilis from Salinas Grandes de Hidalgo (province of La Pampa) and Artemia franciscana from Laguna Mar Chiquita (province of Cordoba), were grown up until adulthood at different salinities (30, 60, 90, 120 gL⁻¹) and temperatures (12, 21, 28°C). The aim was to assess the effects of these different conditions on prereproductive life span and reproductive traits. Results evidenced that at 21 and 28°C, at any salinity, A. franciscana from Laguna Mar Chiquita attained higher survival and fecundity, after a shorter prereproductive period, than A. persimilis from Salinas Grandes de Hidalgo. These data support that A. franciscana, considered a superspecies, exhibits higher phenotypic plasticity than A. persimilis, and that A. persimilis is better adapted to lower temperatures than A. franciscana. These differences in temperature and salinity tolerance could explain the present distribution of these two species in the South Cone in South America. Handling Editor: J. Melack     

International study on Artemia LXIII. Field study of the Artemia urmiana (Günther, 1890) population in Lake Urmiah, Iran

Lake Urmiah is a large (total surface 4750–6100 km² in recent times) thalassohaline hypersaline lake (150–180 g l^–1 in the period 1994–1996), located in northwestern Iran. It is the habitat of the endemic Artemia urmiana. Over the period July 1994–January 1996 a sampling campaign was organized: 36 fixed sampling stations, distributed over the entire lake's area, were sampled weekly to determine water temperature, salinity and transparency. At each occasion a filter net was dragged over a distance of 400 m in the superficial water layer to assess the density and composition of the Artemia population. A more limited sampling campaign focused on the annual fluctuations in chlorophyll concentration and on the reproductive behaviour of the brine shrimp population. Several stages of brine shrimp survived during winter months (water temperature 3°C) at low densities. Compared to available data for the Great Salt Lake, USA, Lake Urmiah shows a low algal biomass and overall low Artemia density. The increasing grazing pressure of the developing brine shrimp population in spring seems to prevent the phytoplankton from reaching high blooming concentrations, and oviparity is the dominant reproductive mode throughout the reproductive season.     

Sediment cores from Lake Urmia (Iran) suggest the inhabitation by parthenogenetic Artemia around 5,000?years ago

In Lake Urmia area, northwestern Iran, parthenogenetic Artemia and the bisexual Artemia urmiana Günther 1890 are found to occupy different ecological niches determined by salinity. Given the fluctuations of the lake over geological times, we thus hypothesized that species identification of Artemia cysts, buried in the sediments, can provide information on lake conditions in the past. Therefore, encysted embryos of Artemia were recovered from lake sediments by augering at a site near the present shoreline. Cysts and associated plant remains from two studied levels yielded radiocarbon ages in the range 5,000–6,700 YBP. For determination of the type of Artemia, the constant synonym mutation in exon-7 of the Na/K ATPase gene was verified, and the diameter of the recovered cysts was compared with that of modern cysts from the Lake Urmia region. The results show that the cysts represent a parthenogenetic type of Artemia, whose cyst diameter is somewhat different from that of present-day local parthenogenetic Artemia. The present study firstly confirms the stability of DNA in ancient Artemia cysts for molecular analysis. Moreover, it suggests variation in Lake Urmia’s conditions over time, and based on comparison with salinity preferences of contemporary Artemia populations, it more specifically suggests that Lake Urmia was a brackish lake dominated by a parthenogenetic Artemia population in the geological period sampled. It finally illustrates how, like in the study of freshwater propagule banks, paleogenetic analysis of Artemia DNA recovered from sediment cores can be used as a tool in the paleoecological study of generally highly fluctuating saline habitats.     

Coexistence of Sexual and Parthenogenetic Artemia Populations in Lake Urmia and Neighbouring Lagoons

We studied the Artemia populations existing in Lake Urmia (north‐western Iran), one of the largest habitats of Artemia in the world, in order to settle the long‐standing controversy over the sexual status of the endemic Artemia populations. Experiments were carried out in the laboratory and in the field. Cysts, collected from different sites of the lake and peripheral lagoons, were hatched and cultured to adults in the laboratory. Adult sexual and parthenogenetic animals were isolated and newly hatched nauplii from them were cultured to maturity in different salinities, ranging from 15–80 ppt. Survival levels and percentage of animals attaining adulthood were measured over a period of 30 days. In the field experiment, cysts taken from Lake Urmia were hatched and the resulting nauplii were inoculated into six earthen ponds (80–140 ppt) constructed in the vicinity of the lake. Population composition in each pond was determined over a period of two years. Results indicated that both sexual and parthenogenetic Artemia coexist in Lake Urmia. While the lake itself is dominated by sexual Artemia, the asexual populations were found to be restricted to particular areas in or near the lake. Artemia appearing seasonally in the lagoons adjacent to the lake were exclusively parthenogenetic. Parthenogens could grow, mature and reproduce at very low salinities (15–33 ppt), whereas higher salinities (above 50 ppt) were required for A. urmiana to attain sexual maturity. We consider salinity to be a major abiotic factor determining the distribution of these sexually different populations within and outside the lake. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Genetic variation and evolutionary origins of parthenogenetic Artemia (Crustacea: Anostraca) with different ploidies

Using two nuclear (ITS1 and Na⁺/K⁺ ATPase) and three mitochondrial (COI, 16S and 12S) markers, we determined the genetic variation and evolutionary relationship of parthenogenetic and bisexual Artemia. Our analyses revealed that mitochondrial genes had higher genetic variation than nuclear genes and that the 16S showed more variety than the other mitochondrial genes in parthenogenetic populations. Triploid parthenogens showed lower genetic variation than diploid ones, whereas the tetra‐ and pentaploids had greater genetic distance than diploid parthenogens. No shared haplotype was found between individuals of parthenogenetic populations and Asian bisexual species with the exception of Na⁺/K⁺ ATPase (Artemia tibetiana). Only mitochondrial markers can demonstrate phylogenetic relationships, and showed that the parthenogenetic Artemia is a polyphyletic group in which the diploid lineages share a common ancestor with Artemia urmiana while tetraploids are closely related to Artemia sinica. The triploid and pentaploid linages are likely to be directly derived from diploid and tetraploid parthenogens, respectively. Subsequently, west Asia is origin for di‐/triploids, and tetra‐/pentaploids rose from East Asia.     

Phylogenetic study of bisexual Artemia using random amplified polymorphic DNA

Study of polymorphisms in the eukaryotic genome is an important way to discover the evolutionary relationships between species. Artemia (Crustacea, Anostraca) offers a very interesting model for evolutionary studies. In fact the genus, distributed all over the world in hundreds of known biotopes, comprises both bisexual sibling species and parthenogenetic populations easily available from the Artemia Reference Center of Ghent. In spite of great interest in it and its extensive use in aquaculture, little is known about relationships between the different species and intraspecific populations. Recently it has been demonstrated that polymorphisms in genomic fingerprints generated by arbitrarily primed polymerase chain reaction (PCR) can distinguish between strains in many organisms. We have used this technique to estimate the phylogenetic relationships existing between 14 populations living in the American continent, in the Mediterranean area, and in China. The principal coordinate analysis (PCO) obtained from 86 random amplified polymorphic DNA (RAPD) markers indicates that the populations analyzed can be divided into homogeneous clusters representing the four known bisexual species—the American A. franciscana and A. persimilis, the Mediterranean A. salina, and the A. species from China.     

Salinity effects on maturation, reproductive and life span characteristics of four Egyptian Artemia populations (International Study on Artemia. LXVIII)

Three parthenogenetic Artemia populations, i.e. two coastal from Borg El-Arab and El-Max saltworks and one from the inland Qarun Lake, and a bisexual strain (Artemia salina) from the inland carbonate lake of Wadi El-Natrun, all from upper Egypt were assayed for their response in 5 salinities (i.e. 35, 80, 120, 150 and 200 g l^?1). The experimental procedure was carried out under laboratory conditions, where the effects of salinity on maturation and ten reproductive and life span characteristics were investigated. The parthenogenetic Egyptian populations are more euryhaline compared to the bisexual one. The two coastal parthenogenetic populations appeared to be very similar in maturation rate and reproductive output at all salinities tested. The inland asexual strain showed a different reproductive response to the elevation of salinity from the two coastal populations. Discriminant function analysis has proven to be a useful tool in determining the differential response of closely related Artemia populations. The bisexual population showed significantly lower reproductive output compared to the parthenogenetic ones and performed best at 35 g l^?1; this is the first record of an A. salina population inhabiting a carbonate lake. These findings may provide valuable information on Artemia biodiversity in an area where very little is known. %     

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.     

Further evidence and characterization of Artemia franciscana (Kellogg, 1906) populations in Argentina

Aim This paper reports the presence of Artemia franciscana ( Kellogg, 1906 ) in Mar Chiquita (CHI) salt lake and in Las Tunas (TUN) lagoon, as well as at a shallow lake in Salinas Grandes (Province of Córdoba) in Argentina. To date, this species has been considered absent from Argentina. This study also provides further data on the characterization of the A. franciscana populations from this area. Location Province of Córdoba (Argentina). Methods The cyst samples collected at the three hypersaline environments were measured to assess its mean diameter with a dissecting microscope. The length of the nauplii hatched from the cysts was also determined with the microscope. The adults raised from these nauplii under strictly controlled conditions were analysed for 12 morphological parameters and compared through multivariate discriminant analysis with other American populations. Fatty acids from the total lipids were analysed by gas chromatography. Reproductive compatibility was evaluated from single‐pair intraspecific and interspecific crosses of adult specimens. Results The results endorse the morphometric assimilation of the Artemia populations of this area to other American populations of the A. franciscana‘super‐species’, together with its morphometric differentiation from the species A. persimilis ( Piccinelli & Prosdocimi, 1968 ). The more conspicuous population from CHI shows cross‐fertility with A. franciscana original from San Francisco Bay (California, USA). Both populations from CHI and TUN show cross‐fertility between them and reproductive isolation with A. persimilis from Hidalgo (Province of La Pampa), which to date is considered to be endemic or exclusive of hypersaline ecosystems in Argentina. The reproductive characteristics displayed by the population from TUN lagoon suggest the existence of an occasional hybridization between A. franciscana and A. persimilis ( Papeschi et al., 2000 ). Artemia cysts from CHI and TUN show a fatty acid profile rich in eicosapentaenoic acid differing markedly from the fatty acid profiles found in A. persimilis cysts. Main conclusions: Artemia franciscana is present in Argentina at 36° S and north of this latitude. Artemia persimilis is confined to the south of latitude 37°10′ S. There are different pieces of evidence that point to a certain level of hybridization of the two species taking place in the land belt between these parallels.     

Artemia monica cyst production and recruitment in Mono Lake,California, USA

Annual egg production was determined for Artemia monica in Mono Lake, California, from 1983 to 1987. Annual oviparous (overwintering cyst) production was 3 and 7 million cysts m^–2 yr^–1 in 1986 and 1987, respectively, as measured by in situ sediment traps. Cyst production for the entire five year period was calculated using Artemia census data and inter-brood duration derived from mixolimnetic temperature. These estimates ranged from 2 to 5 million cysts m^–2 yr^–1. This method underestimated annual production by 30%, when compared to estimates using sediment traps. Cyst production was similar during 1983–1986 and showed a significant increase in 1987, which was due primarily to a larger reproductive population later in the year. Recruitment into the adult populations of the following spring ranged between 1.4 to 3.2%. Overall abundance of this generation reflected the patterns in annual cyst production. Compensatory effects must operate on the second generation of each year, since summer populations were similar in all years despite differences in cyst production.