Oxygen bioavailability and haemoglobins in the brine shrimp Artemia franciscana

Bioavailability of oxygen for the saline-water invertebrate Artemia franciscana was studied, since both oxygen concentration and oxygen diffusion rate change with salinity. Total haemoglobin concentration and the relative contribution of each of three haemoglobins was measured in specimens acclimated to different salinities and oxygen concentrations. Both haemoglobin concentration and contribution were influenced by salinity and the group observed (males, females with and females without eggs). Multiple regression analysis showed that total haemoglobin concentration was better correlated with oxygen concentration than oxygen partial pressure. The relative proportions of haemoglobins 1, 2 and 3 were better correlated with oxygen partial pressure than oxygen concentration. These results are related to the oxygen carrying capacity and oxygen affinity of the haemolymph. Our results show that oxygen bioavailability in a saline-water environment depends on the response (relative contribution of Hb1, Hb2 and Hb3 or total haemoglobin concentration) that is studied. They also show that oxygen concentration and oxygen partial pressure have a different physiological impact on brine shrimp.     

Effect of oxygen and salt on haemolymph oxygen binding in the brine shrimp Artemia franciscana

The oxygen-binding properties of haemolymph from laboratory-reared Artemia franciscana were investigated in vitro. Adult female brine shrimp without eggs were acclimated to seven different combinations of salinity and oxygen. The oxygen affinity (P₅₀) of unprocessed haemolymph rises with acclimation oxygen partial pressure (PO₂) up to normoxic values, after which no influence of oxygen occurs. The increase in P₅₀ with acclimation PO₂ can be explained by the change in proportion of each of three different haemoglobins in the haemolymph. Salinity acclimation has no effect on haemolymph P₅₀. The effects of the different major salts [NaNO₃, NaCl, Ca(NO₃)₂, Mg(NO₃)₂], pH, and the metal-binding ligand EGTA on the oxygen-binding properties of buffered haemolymph (of shrimp acclimated to 10 salinity) were also studied. Little or no effect of these salts could be found. A small Bohr effect (pH 6.5 to 9.1, =-0.11) was noted. Addition of EGTA caused a significant decrease of the oxygen affinity at concentrations up to 50 mmoll^-1.     

Effect of total cadmium and organic complexing on the uptake of cadmium by the brine shrimp,Artemia franciscana

The effect of total cadmium and organic complexing on the rate of cadmium uptake by the brine shrimp, Artemia franciscana has been studied in chemically defined saltwater solutions. The uptake of cadmium from solution by the laboratory-reared brine shrimp displays saturable uptake kinetics. Uptake of cadmium is linear in time up to a total cadmium concentration of 200 moll^-1 and saturates above 800 moll^-1. Complexation of cadmium with organic ligands decreases the uptake of the metal by the brine shrimp. This is in agreement with the view that the availability of cadmium to aquatic organisms is related to the activity of the free cadmium ion in the solution. There is no evidence that the direct uptake of cadmium complexes is important in determining uptake of cadmium. Cadmium uptake is not, however, a mere function of the free cadmium ion activity in the solution, i.e., cadmium uptake rates may differ by an order of magnitude for the same free cadmium ion activity depending on the complexation conditions. In addition to controlling the free cadmium ion activity, the role of organic ligands in metal ion buffering and metal ion masking appear important factors in determining the availability of the metal to the organism.     

Effect of temperature on permeability and drinking rates of the metanauplii of the brine shrimp Artemia sp.

Diffusional permeabilities and drinking rates of 24 h-old Artemia sp. metanauplii incubated at 20 °C were studied at five temperatures: 15, 20, 25, 30, and 35 °C over a period of 6 h. Departures from the incubation temperature resulted in changes in both permeability and drinking rates which both reached their highest values at 35 °C. Metanaupliar permeability and drinking were more elevated than previously reported for adults.     

Relative growth of different species of marine algae in wasterwater-seawater mixtures

In recent studies, we developed a combined nutrient removal-marine aquaculture process for the tertiary treatment of wastewater and the production of commercially important shellfish. Part of this process consists of an outdoor mass cultivation system for marine algae. During our previous experiments we observed that marine diatoms almost exclusively are the dominant algal species in our outdoor cultures. To better understand this phenomenon of diatom dominance we grew 16 species of marine algae in continuous monoculture under laboratory conditions simulating to some degree the conditions prevailing in our outdoor experiments. Species such as Skeletonema costatum, Monochrysis lutheri and Tetraselmis sp., which were never dominant in our outdoor cultures, grew as well in monoculture, as Phaeodactylum tricornutum, frequently, the prevalent species outdoors. However, when monocultures of Dunaliella tertiolecta and Thalassiosira pseudonana (3H) were purposely contaminated with P. tricornutum, the latter species quickly became dominant. It is suggested that a complex interaction of environmental factors is usually responsible for the dominance of a particular species; one such factor may be the nitrogen source in the growth media. Under conditions of virtually, complete nitrogen assimilation, the carbon: nitrogen ratio in the algae was high (7 to 8) when NO ₃ ^- –N was the source of nitrogen, and low (4 to 6) when NH ₄ ⁺ –N was the prime form of nitrogen. When algal growth was low, resulting in a large inorganic nitrogen residue, the carbon:nitrogen ratio was low regardless of whether NO ₃ ^- –N or NH ₄ ⁺ –N was the main nitrogen source.Contribution No. 3297 from the Woods Hole Oceanographic Institution.     

Modulation of the free amino acid pool and protein content in populations of the brine shrimp Artemia spp.

 Free amino acid (FAA) and protein content were measured in various developmental stages of Artemia franciscana, from cysts to Instar III metanauplii. In addition, decapsulated cysts of 15 Artemia populations from different localities were compared with respect to their FAA and protein content. Furthermore, the content and composition of the FAA pool were modulated by hatching the cysts at various salinities, and by enriching the nauplii with algae or a lipid-enrichment emulsion. The FAA content increased threefold from cysts to nauplii, and Instar III metanauplii contained nearly 50% taurine of total FAA. Cysts of A. franciscana were found to contain one-third the amount of FAA compared to the other Artemia species investigated. The content and pool composition of FAA was successfully modulated in 11 of 13 populations, where by the content of FAA was significantly increased when hatched at high salinity. Finally, enrichment elevated the content of FAA and changed the pool composition, thereby showing a dietary effect. Algal enrichment also increased the protein content. Received: 27 September 1999 / Accepted: 17 July 2000     

First report on the triggering effect of light on the hatching mechanism of Artemia salina dry cysts

In the literature, hatching of Artemia salina L. dry cysts has always been related exclusively to the hydration state of the cysts. Hatching experiments with cysts from different places revealed that light in some way triggers the hatching mechanism.     

Relationships between growth rate and RNA,DNA, protein and dry weight in Artemia salina and Euchaeta elongata

The concentrations of RNA, DNA and protein, and the dry weight in 3 cultures of the anostracan Artemia salina (L.) were measured to investigate the usefulness of the RNA-growth relationship in estimating growth or productivity. Similar analyses were performed on Copepodid stages III to VI of the calanoid copepod Euchaeta elongata Esterly collected periodically over 7 months in Haro Strait and over 4 months in Saanich Inlet, British Columbia, Canada. It was found that the RNA-growth relationship derived by Sutcliffe (1965) did not agree with that found for A. salina in this study. Also, neither equation accurately predicted growth rates for E. elongata copepodids. It is concluded that the general positive relationship between RNA concentration and growth rate lacks sufficient specificity to be used as a method for predicting growth rates in these species.     

Light quality in relation to growth,photosynthetic rates and carbon metabolism in two species of marine plankton algae

The effect of light quality on growth, photosynthesis and carbon metabolism in two species of marine algae,Cyclotella nana (Hustedt) andDunaliella tertiolecta (Butcher), was examined. Relative growth constants forC. nana were 0.37, 0.29 and 0.25 in blue, white and green light, respectively. Corresponding constants were 0.41, 0.31 and 0.29 forD. tertiolecta. Photosynthetic rates in both species were higher in blue light and lower in green light compared with white light of the same intensity. More than 60% of¹⁴C assimilated byC. nana orD. tertiolecta grown in blue or green light was incorporated into the ethanol-insoluble fraction, compared with 10 to 30% in this fraction in white light. The relative importance of the various components within this fraction was independent of light quality. Although less¹⁴C was assimilated into the ethanol-soluble fraction in blue or green light, there was a relative increase in some amino acids and organic acids in this fraction and a decrease in sugars and sugar phosphates relative to white light of the same intensity. These differences were independent of light intensity, photosynthetic rate and cell density in the cultures.     

Effect of mercury on the survival,respiration, and growth of postlarval white shrimp,Penaeus setiferus

The results of toxicity experiments have shown that a level of 17 ppb ionic mercury is acutely toxic (96-h LC₅₀) to postlarval white shrimp (Penaeus setiferus). In addition, the size of the shrimp, within the range tested (7 to 35 mm) did not significantly alter the toxicity values of mercury to this species. Fifty-seven days pre-exposure to low levels of mercury (0.5 and 1.0 ppb) had no effect on the acute toxicity (96-h LC₅₀) of mercury to the shrimp. The effect of chronic exposure to low levels of mercury on the respiratory rate, growth, and molting rate of postlarval P. setiferus was studied, and the results showed that 60 days exposure to 0.5 and 1.0 ppb mercury did not significantly affect any of these parameters for the postlarvae.     

The effect of heavy metals on photosynthesis and loss of cell potassium in two species of marine algae,Dunaliella tertiolecta and Phaeodactylum tricornutum

Observations on sixth-stage nauplius (pre-cyprid) of the barnacle Balanus eberneus Gould treated with L-proline-3,4-H³ suggest that the cement gland in this stage is a functional organ with considerable biosynthetic activity; the activity of the cement glands in the cyprid similarly treated is relatively low, ceasing gradually during, and completely after, metamorphosis. In the early phases of this stage, the glands become highly vacuolated (degenerate), and disappear completely in 3 days. The origin of the cement apparatus in the newly metamorphosed barnacle is discussed.     

Effects of temperature and delayed feeding on growth and survival of larvae of three species of subtropical marine fishes

In larvae of the bay anchovy Anchoa mitchilli (Valenciennes), the sea bream Archosargus rhomboidalis (Linnaeus), and the lined sole Achirus lineatus (Linnaeus), growth, survival, and starvation times were investigated at temperatures of 22° to 32°C. The rate at which hours after hatching until starvation decreased in relation to temperature for unfed larvae did not differ significantly among the 3 species, ranging from-5.4 to-6.3 h per degree increase in temperature. The total number of hours until starvation did differ for all 3 species: lined soles survived longest, bay anchovies were intermediate, and sea bream survived the least time. At 28°C, unfed sea bream could survive 90.1 h, bay anchovy 102.3 h, and lined sole 119.8 h. The eyes pigmented at nearly the same time after hatching for sea bream and bay anchovy, but took about 20 h longer at all temperatures for lined sole. Quadratic equations best described the relationship between hours after hatching when the eyes became pigmented and temperature. Eye-pigmentation times became nearly constant for all 3 species at temperatures above 28°C. At 28°C, eyes pigmented about 27 h after hatching for bay anchovy and sea bream but not until 47 h for lined sole. Hours after eye pigmentation when unfed larvae starved was a measure of the effective time that larvae had to commence feeding. Bay anchovies and lined soles were nearly alike in this respect, but sea bream starved at tewer hours after eye pigmentation. Slopes of regressions representing decrease in times to staration for increasing temperatures ranged from-3.7 to-4.4 h per degree increase in temperature, and were not significantly different among the 3 species. At 28°C, unfed lined soles starved at 70 a after eye pigmentation, bay anchovies starved at 72.5 h, and sea bream at only 62 h. Yolk absorption was most rapid for all species during the first 20 h after hatching, and was faster at higher temperatures. Amounts of yolk remaining at the time eyes became pigmented were less at higher temperatures for bay anchovy and lined sole, but were greater for sea bream, suggesting that sea bream used yolk more efficiently at higher temperatures. Either no yolk or small traces (>0.20%) remained at 24 h after eye pigmentation in all 3 species. Feeding was delayed for periods of 8, 16, 24, 32, 40 and 48 h after eye pigmentation for all species at a series of experimental temperatures from 24° to 32°C. Growth and survival were affected when food was withheld for more than 24 h at 28°C, but survival did not decrease markedly until food was withheld at least 8 h longer. At lower temperatures food could be withheld longer and at higher temperatures for less time. Feeding can be initiated by most larvae for several hours after all visible yolk reserves have been exhausted. All species tested can survive for 24 to 40 h after eye pigmentation at 24° to 28°C without food and still have relatively good growth and survival when food is offered. If the “critical period” is considered relative to time of hatching, lined soles need not find food for 3 to 3.5 days after hatching, but bay anchovy and sea bream must feed within 2.5 days of hatching.     

Effect of biosolid hydrochar on toxicity to earthworms and brine shrimp

The hydrothermal carbonization of sewage sludge has been studied as an alternative technique for the conversion of sewage sludge into value-added products, such as soil amendments. We tested the toxicity of biosolid hydrochar (Sewchar) to earthworms. Additionally, the toxicity of Sewchar process water filtrate with and without pH adjustment was assessed, using brine shrimps as a model organism. For a Sewchar application of 40 Mg ha^?1, the earthworms significantly preferred the side of the vessel with the reference soil (control) over side of the vessel with the Sewchar treatments. There was no acute toxicity of Sewchar to earthworms within the studied concentration range (up to 80 Mg ha^?1). Regarding the Sewchar process water filtrate, the median lethal concentration (LC50) to the shrimps was 8.1% for the treatments in which the pH was not adjusted and 54.8% for the treatments in which the pH was adjusted to 8.5. The lethality to the shrimps significantly increased as the amount of Sewchar process water filtrate increased. In the future, specific toxic substances in Sewchar and its process water filtrate, as well as their interactions with soil properties and their impacts on organisms, should be elucidated. Additionally, it should be identified whether the amount of the toxic compounds satisfies the corresponding legal requirements for the safe application of Sewchar and its process water filtrate.     

Studies on the growth of some marine unicellular algae under different artificial light sources

Documented evidence of the suitability of various artificial light sources for the culture of unicellular algae is scarce. This paper reports the results of a comparison of the growth rates of 5 species of marine unicellular algae at various light intensities produced by an incandescent filament source or daylight fluorescent tubes. Under light saturating conditions the maximum growth rate achieved by each species was the same under each artificial source. Under sub-optimal intensities, no difference in the growth rates under the 2 sources was detected in the case of Phaeodactylum tricornutum, Chlorella ovalis or Monochrysis lutheri. However, under these conditions both Brachiomonas submarina and Dunaliella primolecta required higher intensities of incident illumination from the fluorescent source than from the incandescent source to maintain a particular relative growth rate.     

长江口南支沉积物对卤虫的毒性效应研究

以Ⅱ-Ⅲ龄卤虫幼体（Artemia salina）为受试生物,开展生物检测试验,研究长江口南支沉积物的毒性效应。生物检测结果表明长江口南支部分站位的沉积物对卤虫幼体已产生毒性效应,其中,2、5、8三站沉积物对卤虫体长和体内SOD酶活性具有显著影响（p〈0.05）,1号和2号站沉积物对卤虫体内LDH活性有显著影响（p〈0.05）。对多个毒性效应指标运用因子分析法得到长江口南支各站因子得分F,F值大小可表征各站沉积物对卤虫综合毒性的强弱,各站F值排序为：2号（-1.59）〈5号（-0.76）〈8号（-0.42）〈1号（0.06）〈6号（0.62）〈7号（0.78）〈3号（1.31）,2号站沉积物毒性最强,3号站毒性最弱。经逐步回归分析,因子得分F与沉积物中Cu、Cd含量具有显著相关关系（R=0.91,p〈0.05）,表明长江口南支沉积物中Cu和Cd是对卤虫产生毒性效应的关键污染物。     

Chemical composition of dietary species of marine unicellular algae and rotifers with emphasis on fatty acids

The lipid profiles of a few species of marine unicellular algae and yeast were studied with emphasis on fatty acids as part of a search for the nutritional value of plankton to the diet of marine fish larvae commonly used in marine hatcheries. The general proximate chemistry of rotifers was closely related to the proximate chemistry of the diet organism, exhibiting a higher content of protein and carbohydrate and a lower content of lipid. Major lipids in all algae, yeast and rotifers comprised mono-, di- and tri-glycerides and polar lipids. The algae Chaetoceros gracilis Schutt, Isochrysis galbana Parke and their respective algaefed rotifers exhibited higher amounts of neutral lipids, consisting mainly of cyclic and branched polyunsaturated components. Fatty acid composition of the algae was species-specific, with the highest ratio of polyethylenic to saturated and monoethylenic acid in I. galbana and Phaeodactylum tricornutum Bohlin, and the highest content (15%) of n-3 highly unsaturated fatty acids in Nannochloropsis salina and P. tricornutum. A closely mirrored distribution of the fatty acids, but with a lower amount of n-3 highly unsaturated fatty acids, was present in the respective algae-fed rotifers. Comparison of the fatty acid spectrum of Artemia sp. and Euterpina acutifrons grown in the laboratory on I. galbana with zooplankton samples of E. acutifrons and Oitona nana collected from the sea showed a higher concentration of docosahexaenoic acid (22:6 n-3) in the naturally collected sample. The results indicate that the efficacy of the food algae C. gracilis and I. galbana in increasing the survival of fish larvae in marine hatcheries is not obvious on the sole basis of fatty acid composition.     

Effects of ammonia on survival and osmoregulation of various development stages of the shrimp Penaeus japonicus

In Penaeus japonicus, the tolerance to ammonia increased with the development from nauplius to late juvenile. The 48-h LC₅₀ of ammonia in nauplii (III–V), 96-h LC₅₀ in zoeae (I–III), mysis (I–III), post-larvae (PL1) and late juveniles (10.4±1.1 g) were respectively 5.0, 6.1 to 8.1, 9.4 to 10.9, 15.5 and 52.7 mg Nl^-1 (0.5, 0.6 to 0.7, 0.9, 1.3 and 3.1 mg NH₃–Nl^-1). In a chronic experiment (20 d), the LC₅₀ in post-larvae (PL1) was 19.1 (1.4) at 96 h and 16.2 mg Nl^-1 (1.3 mg NH₃–Nl^-1) at 480 h. Osmoregulatory capacity (OC) was calculated as the osmotic gradient between the hemolymph and the external medium at given salinities. The effects of ammonia on OC, Na⁺ and Cl^- regulation and gill Na⁺–K⁺ ATPase activity in late juveniles were examined in fullstrength seawater, SW (1050 mosm kg^-1, 36 S) and in dilute SW (450 mosm kg^-1, 15%.), after 48 or 96 h exposure to various concentrations of ammonia. Ambient ammonia disrupted both hypo- and hyper-osmoregulation; decreased OC resulted from impaired Na⁺ and Cl^- regulation. Gill Na⁺–K⁺ ATPase activity increased in SW and was not affected in dilute SW. The decrease of OC was ammonia-dose-dependent. The threshold ammonia concentrations affecting hypo-OC and hyper-OC were, respectively, 16 (1.3) and 32 mg Nl^-1 (2.3 NH₃–Nl^-1) for a 48 h exposure; these concentrations were lower than the 48-h LC₅₀ value, 65.3 mg Nl^-1 (3.5 NH₃–Nl^-1). The time course of exposure to sublethal ammonia (48 mg Nl^-1) demonstrated that the effect on osmoregulation was time-dependent. This effect was also temporary, and the exposed shrimps recovered control OC values after removal of excessive ambient ammonia. The possibility of using OC as an indicator of physiological condition in osmoregulating crustaceans and the acting mode of ammonia on osmotic and ionic regulation are discussed.     

Light quality and concentration of proteins,RNA, DNA and photosynthetic pigments in two species of marine plankton algae

Chlorophyll, protein, DNA and RNA concentrations in cultures ofCyclotella nana (Hustedt) andDunaliella tertiolecta (Butcher) were higher in blue light and lower in green light than in white light of the same intensity. Total carotenoid concentrations were highest in green light. Total pigment concentrations were highest in cells grown in blue light, lowest in those from green light. The relative changes in pigment concentrations associated with differences in spectral composition of the radiation are comparable to those often observed in phytoplankton from various depths in stratified natural water. Light adaptation in algae as a response to light quality is suggested. The chemical composition of plankton algae can be expected to vary considerably with depth.     

Effects of UV-B irradiation on growth and survival of Antarctic marine diatoms

Growth rate, survival, and stimulation of the production of UV-B (280 to 320 nm) absorbing compounds were investigated in cultures of five commonly occurring Antarctic marine diatoms exposed to a range of UV-B irradiances. Experimental UV-B exposures ranged from 20 to 650% of the measured peak surface irradiance at an Antarctic coastal site (0.533 J m^-2 s^-1). The five diatom species (Nitzschia lecointei, Proboscia alata, P. inermis, Thalassiosira tumida and Stellarima microtrias) appear capable of surviving two to four times this irradiance. In contrast to Phaeocystis cf. pouchetii, another major component of the Antarctic phytoplankton, the concentrations of pigments with discrete UV absorption peaks in diatoms were low and did not change significantly under increasing UV-B irradiance. Absorbance of UV-B by cells from which pigments had been extracted commonly greatly exceeded that of the pigments themselves. Most of this absorbance was due to oxidisable cell contents, with the frustule providing the remainder. Survival of diatoms did not correlate with absorption by either pigments, frustules or oxidisable cell contents, indicating that their survival under elevated UV-B irradiances results from processes other than screening mechanisms.     

铜尾矿对5种豆科植物根系生长的影响

针对铜陵市铜尾矿废弃地的复垦，选用了5种乡土豆科植物在5种不同的尾矿改良方式上进行盆栽试验。5种供试物种为：大豆(Glycine max)、赤豆(Phaseolus angularis)、赤小豆(P.calcaratus)、绿豆(P.radiatus)、山绿豆(P.mininus)。盆栽基质是以尾矿与正常土壤按体积100：0、75：25、50：50、25：75、0：100比例混合而成，分别标记为：TAl00、TA75、TA50、TA25、TA0005种豆科植物生长时间为70d。结果发现：随铜尾矿在混合基质中比例的增加，5种豆科植物根系的主根长逐渐变短、根系一级侧根的数目逐渐减少；铜尾矿对5种豆科植物根瘤的形成产生抑制作用，特别是山绿豆和绿豆在全尾矿的基质上，无根瘤生成；地下根部的生物量表现为随着尾矿含量比例的增加。地下部分的生物量逐渐下降，但根冠比表现为随尾矿含量比例的增加而增加；植物对铜尾矿中铜的积累，主要含集中根部，地上部分与地下部分差异明显。