Acid brine shrimp: Metabolic strategies in osmotic and ionic adaptation

Acid salt lakes are found in several regions of Australia but are uncommonly abundant in the Yilgarn Block area of southwestern Australia. The chemical properties of the acid salt lakes are in general similar to those of shallow ephemeral alkaline salt lakes found in adjacent regions except for having a higher hydrogen ion concentration and an absence of carbonate and bicarbonate ions. The Australian brine shrimp, Parartemia, is the major zooplankter living in these salt lakes. Present investigation on two species of larval Parartemia reveal P. zietziana nauplii having high salt tolerance (LD₅₀ > 225 TDS) but a narrow pH range with an optimum lying near pH 8. In contrast, P. contracta collected from acid salt lakes had a more restricted salt tolerance (LD₅₀ < 100 TDS) but a wider range of pH tolerances with substantial survival below pH 3.5. Both species, P. zietziana and contracta, when placed in ouabain-laden salines, demonstrated decreased survival and indirectly indicated the presence of a ouabain-sensitive sodium pump. Direct enzymatic assay of the sodium pump (Na, K ATPase) in nauplii of P. zietziana gave specific activity values of 2.9 µM Pi/hr/mg protein supporting our working hypothesis that the nauplii of Parartermia have an osmoregulatory system similar to that found in larval Artemia which is dependent upon having large quantities of ATP to support the sodium pump. In Artemia larvae, the production of ATP is enhanced through a facultative pathway involving an aerobic glycolysis linked C-4 dicarboxylic shunt. The major CO₂ source for the C-4 acid shunt for alkaline brine shrimp has been found to be the dissolved bicarbonate/carbonate ions. In highly acidic saline lakes, these ions are missing. If acid brine shrimp are to survive in low pH ephemeral saline lakes, they must have evolved an additional proton pump and devised a mechanism to produce ATP from endogenous CO₂ substrates.     

Are crustaceans linked to the ciguatera food chain?

Synopsis Adult brine shrimp, Artemia spp., were used as an experimental organism to elucidate the role that crustacea may play in the transference of ciguatera toxins. Some ciguatera-implicated dinoflagellates were highly toxic to brine shrimp that had consumed them. Four clones of Gambierdiscus toxicus were fed in four trials at rates ranging from 2 to 480 cells per adult brine shrimp; the 24 h LD₅₀ for the four clones were 2.8, 33.4, 41.1, and 104.5 cells per brine shrimp. Dinoflagellates Prorocentrum concavum and P. lima were also fed to adult brine shrimp, but minimal mortalities occurred at cell concentrations ranging up to 1000 cells per test animal. Tilapine cichlid (Oreochromis niloticus ×O. mossambicus) young fed brine shrimp containing G. toxicus cells displayed behavioral abnormalities ranging from spiral swimming to loss of equilibrium. The present data suggest that toxins accumulated by dinoflagellate-consuming crustaceans could produce toxicity in zooplanktivorous fish species, or to detritivores in cases where dinoflagellate consumption resulted in crustacean mortalities. Field studies of the ciguatera food chain should be expanded to include examination of crustacean diets to more fully define their role in toxin transfer.     

Abundance and spatial distribution of Artemia salina in Lake Abert,Oregon

In 1981–1982 Abert Lake had an area of 200 km², with a mean depth of 2.5 m and a total dissolved salt concentration of 82 gl^-1. The spatial distribution and abundance of the brine shrimp, Artemia salina, were monitored for 1981–1982. In 1981 during the midsummer months, with sampling primarily in the eastern side of Abert Lake, brine shrimp populations showed peak densities of 2–4 ind. 1^-1. In 1982, peak abundance occurred in early July and ranged between 5–8 ind. 1^-1. Lakewide estimates of brine shrimp derived from 14 collecting stations and assuming a uniform distribution over the lake resulted in estimates of lakewide abundance of 3 × 10¹¹ adults with a total biomass of 7 × 10⁶ kg. The major algal species in the lake was a benthic filamentous green alga (Ctenocladus circinnatus).     

Functional morphology of the neck organ in Artemia salina nauplii

Fine structure of the ion transporting epithelium of the neck organ in the brine shrimp (Artemia salina) nauplius is described. The neck organ is a dome-like gland situated atop the cephalothorax of the larva and is composed of 50 to 60 cuboidal epithelial cells. These cells possess many of the characteristics of salt-secretory cells from other tissues. They contain many mitochondria and exhibit a high degree of plasma membrane elaboration. This membrane amplification takes two forms; the apical plasmalemma is infolded into irregular loops, while the basal and lateral membranes penetrate the cytoplasm in the form of branching sinusoids. The labyrinth of tubular reticulum thus formed fills most of the cell volume. Mitochondria in the labyrinth are often in intimate contact with these tubular membranes and regular arrays of parallel mitochondria with constricted intervening sinusoids are often observed. Other organelles including Golgi complexes, multivesicular bodies, and rough endoplasmic reticulum are also numerous, particularly in the narrow rim of cytoplasm which lies between the apical infolds and the labyrinth. Yolk platelets and glycogen fields are conspicuous in the basal perinuclear regions of the cells.     

The American brine shrimp as an exotic invasive species in the western Mediterranean

The hypersaline environments and salterns present in the western Mediterranean region (including Italy, southern France, the Iberian Peninsula and Morocco) contain autochthonous forms of the brine shrimp Artemia, with parthenogenetic diploid and tetraploid strains coexisting with the bisexual species A. salina. Introduced populations of the American brine shrimp A. franciscana have also been recorded in these Mediterranean environments since the 1980s. Based on brine shrimp cyst samples collected in these countries from 1980 until 2002, we were able to establish the present distribution of autochthonous brine shrimps and of A. franciscana, which is shown to be an expanding invasive species. The results obtained show that A. franciscana is now the dominant Artemia species in Portuguese salterns, along the French Mediterranean coast and in Cadiz bay (Spain). Co-occurrence of autochthonous (parthenogenetic) and American brine shrimp populations was observed in Morocco (Mar Chica) and France (Aigues Mortes), whereas A. franciscana was not found in Italian cyst samples. The results suggest these exotic A. franciscana populations originate as intentional or non-intentional inoculations through aquacultural (hatchery effluents) or pet market activities, and suggest that the native species can be rapidly replaced by the exotic species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Energy Budgets for Eared Grebes on the Great Salt Lake and Implications for Harvest of Brine Shrimp

ABSTRACT About 1.5-million eared grebes (Podiceps nigricollis), representing half of the North American population, stage on Utah's Great Salt Lake, USA (GSL) during autumn migration to forage on brine shrimp (Artemia franciscana). Indirectly competing with birds for brine shrimp are commercial harvesters who annually collect >1 million kg (dry wt) of shrimp cysts (i.e., hardened eggs), an amount that during some years equals up to half of all brine shrimp cysts produced annually on the GSL. No information was available regarding what impact this commercial harvest was having on eared grebes. We determined daily energy requirements of eared grebes so that regulations governing brine shrimp cyst harvest would better reflect foraging needs of grebes. We measured basal metabolic rate (BMR) of eared grebes from June 2000 to October 2000. Mean BMR of 106 adult and subadult eared grebes was 0.023 kJ/g/hour (SD = 0.004), and mean BMR of 37 juveniles was 0.024 kJ/g/hour (SD = 0.003). Resting and preening metabolic rates were 1.2 times higher than BMR, whereas diving-bout metabolic rate was 1.7 times higher than BMR. Daily energy needs of an average-sized grebe (550 g) during November were 391 kJ. Meeting this energy need requires daily consumption of 24,400 adult brine shrimp. In addition, grebes must consume 2,100–5,200 adult shrimp daily to obtain enough energy reserves to continue their migration to California, USA, and Mexico. Hence, grebes need to consume 26,500–29,600 adult brine shrimp daily while staging on GSL. To achieve this high harvest rate, grebes need adult brine shrimp densities at >380 shrimp/m³during autumn. Commercial harvest of brine shrimp cysts from GSL should be curtailed when cyst densities fall below 20,000 cysts/m³to ensure enough adult brine shrimp for grebes during the subsequent year.     

Effect of water-borne copper on respiratory and cardiac function during the early ontogeny of the brine shrimp,Artemia franciscana Kellogg 1908 (Branchiopoda: Anostraca)

There was no direct effect of copper on the ontogeny or function of the heart of the brine shrimp Artemia franciscana in sea water (salinity= 36 mg·ml^-1, 25°C). There was, however, an indirect effect as an increase in copper concentration resulted in a reduced growth rate. There was no difference between the critical O₂ tensions of newly hatched (stage 0/1) nauplii of control and treated (<0.32 and 10.11 mol·l^-1 copper, respectively) individuals. However by developmental stages 4–6, when both the heart and thoracic gills are in the process of differentiating, respiratory performance had improved (i.e. critical O₂ tension decreased from 6.27±0.45 to 4.69±0.24 kPa) in control but not in copper-treated individuals. It is suggested that respiratory impairment of stages 4–6 individuals is unlikely to be related to differences in cardiac performance or cellular respiration. Instead it may be related to metal-related damage to newly differentiating gill tissue and/or by copper in some way compromising the normal ontogenic shift in haemoglobin O₂ affinity. Copper-related respiratory impairment develops at a critical point in brine shrimp organogenesis when a good supply of O₂ is essential for normal development and if compromised may reduce the ability of this species to survive copper exposure.Abbreviations BL body length - BW body weight - HR heart rate - HM heavy metals - SW sea water - P _c critical oxygen tension     

有毒亚历山大藻对卤虫存活率和摄食率的影响

研究了有毒亚历山大藻对卤虫存活率和摄食率两方面的影响,得出以下结论：在卤虫存活率实验中,有毒亚历山大藻在2000cells／ml的密度下,对卤虫具有致死效应,卤虫在24-168h内全部死亡;在摄食实验中,有毒亚历山大藻对卤虫的摄食产生明显的抑制作用,卤虫对有毒藻的平均摄食率明显低于无毒藻组和混合实验组。在加入无毒藻东海原甲藻的混合培养状态下。卤虫存活率上升,30-60min摄食率增加,东海原甲藻在一定程度上可以减轻塔玛亚历山大藻对卤虫的毒害作用。有毒藻产生的PSP毒素并非导致卤虫死亡的主要原因,毒害作用可能与出现在卤虫体外的黏附物质有关。通过对3个不同生长期卤虫的研究发现,后无节幼体卤虫对有毒亚历山大藻的毒害作用最为敏感。     

Nearshore and pelagic abundances of Artemia monica in Mono Lake,California

The spatial distribution and abundance of the brine shrimp, Artemia monica, in Mono Lake, California were determined during 1982 and 1983. Peak abundances of shrimp occur in midsummer and reach densities of 15–17 individuals l^-1 in the nearshore regions and 6–8 individuals 1^-1 in the pelagic region. The brine shrimp were non-uniformly distributed both vertically and horizontally. The coefficient of variation in shrimp abundance among stations within the nearshore region was similar to that found in the pelagic region. On two of the nine dates, nearshore densities were 3 to 4 times greater than those in the pelagic zone, and on average the brine shrimp appear to be slightly over-dispersed to the nearshore region. However, including nearshore abundances in lakewide estimates will usually result in a change of less than a 10%.     

Molecular cloning and expression analysis of the interferon-γ-inducible lysosomal thiol reductase gene from the shrimp <Emphasis Type="Italic">Penaeus monodon</Emphasis>

The interferon-γ-inducible lysosomal thiol reductase enzymes (GILT) have been shown to play an important role in the processing of exogenous antigens by catalyzing disulfide bond reduction, that facilitates unfolding of the native protein antigen to simplify further cleavage by cellular proteases. In this study a Penaeus monodon GILT (PmGILT) gene was isolated from an EST library of white spot syndrome virus (WSSV)-infected P. monodon. The full-length cDNA of the PmGILT gene was 780 bp and contained an open reading frame of 657 bp that encoded 218 amino acid residues with a predicted protein molecular weight of 24 kDa. The deduced amino acid sequence of PmGILT contains an active site CXXS motif, a GILT signature sequence (CQHGX₂ECX₂NX₄C) and 10 conserved cysteines together with other signature characteristics of GILT proteins. RT-PCR analysis showed that the PmGILT mRNA expression level was clearly up-regulated in the lymphoid organ of both the LPS-induced and WSSV-infected shrimp, compared to normal shrimp. In response to WSSV infection, the penaeid shrimp JAK/STAT pathway is reported to play an important role in the lymphoid organ. We hypothesize that this activated STAT may stimulate GILT expression so that it can be involved in the shrimp immune response system.     

Complementation analysis of the inorganic carbon concentrating mechanism of Chlamydomonas reinhardtii

Summary Six independently isolated mutants of Chlamydomonas reinhardtii that require elevated CO₂ for photoautotrophic growth were tested by complementation analysis. These mutants are likely to be defective in some aspect of the algal concentrating mechanism for inorganic carbon as they exhibit CO₂ fixation and inorganic carbon accumulation properties different from the wild-type. Four of the six mutants defined a single complementation group and appear to be defective in an intracellular carbonic anhydrase. The other two mutations represent two additional complementation groups.Abbreviations HS high salt medium which has 13 mM phosphate at pH 6.8 - HSA high salt plus 36 mM acetate medium - YA high salt medium with 4 g yeast extract per L and 36mM acetate - Arg arginine - cia^- CO₂ accumulation mutants that cannot grow on low CO₂ - C_i inorganic carbon (CO₂+HCO _- ³ ) - CA carbonic anhydrase - mt mating type Supported in part by the McKnight Foundation and by NSF grant PCM 8005917 and published as journal article 11924 from the Michigan State Agriculatural Experiment Station     

Surface specialization of the larval salt gland inArtemia salina nauplii

Summary Scanning electron microscopic (SEM) observations of brine shrimp nauplii reveal the dorsal surface of the cephalothorax region to be specialized in structure. During larval differentiation the epithelial layer is transformed from a flat caplike structure into a hemispherical dome that has deep channels coursing through the epithelium. A thin cuticular film covers the epithelial cap but a distinct ruffled edge serves as a boundary that identifies the limits of the organ. These morphological observations support previous finding which indicated this area to be the site of the larval salt gland.This research was supported by AEC grant RLO 2227-T13-2. One of the authors (FPC) wishes to thank Mr. Michael Nemanic, Dept. of Zoology, University of California for help in preparing specimens and to Dr. Thomas Hayes, Donner Lab, for the generous use of his scanning electron microscope. The authors also wish to thank Dr. Franco Rossi, Mr. Charles Dwire, and Mrs. Joan Dwire of Electros Analytics, A Division of Electros, Inc. Tigard, Oregon for the time and effort given in the preparation of the micrographs of the freeze-dried specimens.     

DISTINCTIVE LOCALIZATION OF GROUP 3 LATE EMBRYOGENESIS ABUNDANT SYNTHESIZING CELLS DURING BRINE SHRIMP DEVELOPMENT

Despite numerous studies on late embryogenesis abundant (LEA) proteins, their functions, roles, and localizations during developmental stages in arthropods remain unknown. LEA proteins protect crucial proteins against osmotic stress during the development and growth of various organisms. Thus, in this study, fluorescence in situ hybridization was used to determine the crucial regions protected against osmotic stress as well as the distinctive localization of group 3 (G3) LEA⁺ cells during brine shrimp development. Several cell types were found to synthesize G3 LEA RNA, including neurons, muscular cells, APH‐1⁺ cells, and renal cells. The G3 LEA⁺ neuronal cell bodies outside of the mushroom body projected their axonal bundles to the central body, but those inside the mushroom body projected their axonal bundles toward the deutocerebrum without innervating the central body. The cell bodies inside the mushroom body received axons of the G3 LEA⁺ sensory cells at the medial ventral cup of the nauplius eye. Several glands were found to synthesize G3 LEA RNA during the nauplius stages of brine shrimp, including the sinus, antennal I and II, salt, and three ectodermal glands. This study provides the first demonstration of the formation of G3 LEA⁺ sinus glands at the emergence stages of brine shrimp. These results suggest that G3 LEA protein is synthesized in several cell types. In particular, specific glands play crucial roles during the emergence and nauplius stages of brine shrimp.     

Luminescence, virulence and quorum sensing signal production by pathogenic Vibrio campbellii and Vibrio harveyi isolates

Aims:  To study the relationship between luminescence, autoinducer production and virulence of pathogenic vibrios. Methods and Results:  Luminescence, quorum sensing signal production and virulence towards brine shrimp nauplii of 13 Vibrio campbellii and Vibrio harveyi strains were studied. Although only two of the tested strains were brightly luminescent, all of them were shown to produce the three different types of quorum sensing signals known to be produced by Vibrio harveyi. Cell-free culture fluids of all strains significantly induced bioluminescence in the cholerae autoinducer 1, autoinducer 2 and harveyi autoinducer 1 reporter strains JAF375, JMH597 and JMH612, respectively. There was no relation between luminescence and signal production and virulence towards brine shrimp. Conclusions:  There is a large difference between different strains of Vibrio campbellii and Vibrio harveyi with respect to bioluminescence. However, this is not reflected in signal production and virulence towards gnotobiotic brine shrimp. Moreover, there seems to be no relation between quorum sensing signal production and virulence towards brine shrimp. Significance and Impact of the Study:  The results presented here indicate that strains that are most brightly luminescent are not necessarily the most virulent ones and that the lower virulence of some of the strains is not due to a lack of autoinducer production.     

Localization patterns of dopamine active transporter synthesizing cells during development of brine shrimp

There have been many studies on dopamine active transporter (DAT) in humans and laboratory animals; however, there is a lack of information on DAT in brine shrimp. In this study, we demonstrated the neuronal and nonneuronal characteristics of DAT‐synthesizing (DAT⁺ cells) during development of brine shrimp. In neuronal cells, the DAT⁺ neurons in the central body and lobes of a protocerebrum (PC) controlled the deutocerebrum. The sensory cells of nauplius eyes projected their decussated axons to the PC, and the DAT⁺ cells at the posterior region were associated with migration and control of the 10 posterior neurons during the early nauplius stage. In nonneuronal cells, the five types of glands, that is, the salt, antennal, mandible, and accessory glands and posterior gland1 and gland2 synthesized DAT protein. In addition, the gut and rectum dilator muscles and renal cells expressed DAT protein. Thus, DAT protein acts in the development of several types of cells during development of brine shrimp.     

Optimum growth conditions for Amphidinium carterae Hulburt from eutrophic waters in Alexandria (Egypt) and its toxicity to the brine shrimp Artemia salina

Cultures of a strain of Amphidinium carterae Hulburt isolated from a eutrophic basin in Alexandria were investigated for the optimum growth conditions, including vitamin requirements, the assimilation index and the potential toxicity of this species. A division rate of 2.7 day^–1, was reached on the second and third day under optimum conditions, with and without addition of B₁₂ and biotin. The rate of C₁₄ assimilation per unit chlorophyll ranged from 0.2 to 5.4 mgC.(mg Chla)-1.h^–1 according to the growth conditions and to cell size. As the cell density increased in the cultures, the cell size decreased and their photosynthetic efficiency increased. Toxicity of A. carterae to brine shrimp resulted from grazing and not from membrane diffusion of extracellular toxins.     

Indole analogues decreasing the virulence of Vibrio campbellii towards brine shrimp larvae

Indole signalling has been proposed as a potential target for the development of novel virulence inhibitors to control bacterial infections. However, the major structural features of indole analogues that govern antivirulence activity remain unexplored. Therefore, we investigated the impact of 26 indole analogues on indole-regulated virulence phenotypes in Vibrio campbellii and on the virulence of the bacterium in a gnotobiotic brine shrimp model. The results demonstrated that 10 indole analogues significantly increased the fluorescence of indole reporter strain Vibrio cholerae S9149, 21 of them decreased the swimming motility of V. campbellii, and 13 of them significantly decreased the biofilm formation of V. campbellii. Further, we found that 1-methylindole, indene, 2,3-benzofuran, thianaphthene, indole-3-acetonitrile, methyl indole-3-carboxylate, 3-methylindole, and indole-2-carboxaldehyde exhibited a significant protective effect on brine shrimp larvae against V. campbellii infection, resulting in survival rates of challenged brine shrimp above 80%. The highest survival of shrimp larvae (98%) was obtained with indole-3-acetonitrile, even at a relatively low concentration of 20 μM. Importantly, the indole analogues did not affect bacterial growth, both in vitro and in vivo. These results indicate the potential of indole analogues in applications aiming at the protection of shrimp from vibriosis.     

The application of bioflocs technology to protect brine shrimp (Artemia franciscana) from pathogenic Vibrio harveyi

Aims: To study the potential biocontrol activity of bioflocs technology. Methods and Results: Glycerol‐grown bioflocs were investigated for their antimicrobial and antipathogenic properties against the opportunistic pathogen Vibrio harveyi. The bioflocs did not produce growth‐inhibitory substances. However, bioflocs and biofloc supernatants decreased quorum sensing‐regulated bioluminescence of V. harveyi. This suggested that the bioflocs had biocontrol activity against this pathogen because quorum sensing regulates virulence of vibrios towards different hosts. Interestingly, the addition of live bioflocs significantly increased the survival of gnotobiotic brine shrimp (Artemia franciscana) larvae challenged to V. harveyi. Conclusions: Bioflocs grown on glycerol as carbon source inhibit quorum sensing‐regulated bioluminescence in V. harveyi and protect brine shrimp larvae from vibriosis. Significance and Impact of the Study: The results presented in this study indicate that in addition to water quality control and in situ feed production, bioflocs technology could help in controlling bacterial infections within the aquaculture pond.     

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.

     

Artemia cysts as an alternative food for the predatory bug Macrolophus pygmaeus

The suitability of cysts of the brine shrimp Artemia sp. as a factitious food for the predator Macrolophus pygmaeus Rambur was investigated. The influence of decapsulation time and hydration of the cysts on the performance of the predator were studied in the absence of plant material. A longer time of decapsulation had a positive influence on the development of the predator. Hydration of cysts had a significant impact on nymphal survival when cysts where non‐decapsulated or poorly decapsulated. An experiment in which nymphs were switched from a diet of hydrated cysts to non‐hydrated cysts showed that in the absence of plant material the relative importance of hydrating the cysts decreased with nymphal age. Especially, the first instar and to a lesser extent the second instar appear to be susceptible to water shortage. Effects of prolonged rearing on development and reproduction on brine shrimp cysts from different origins were tested in the presence of plant material. Rearing M. pygmaeus on Artemia sp. (Jingyu Lake) cysts yielded similar survival, development, adult weight and fecundity in the fourth as in the second generation. In contrast, for Artemia franciscana cysts, an increase in nymphal development was notable. Biochemical analyses showed that total amino acid content and the concentration of the different amino acids did not differ among diets and generations. There were, however, differences in total fatty acid content between the different diets and generations and in the concentration of certain fatty acids, indicating that insects fed brine shrimp cysts may show nutritional deficiencies compared to those reared on a diet of Ephestia kuehniella eggs. Our results indicate that decapsulated brine shrimp cysts are an economically viable alternative food source in at least part of the rearing process for M. pygmaeus.