Growth,salinity tolerance and extracellular carbohydrate yield of Prasinocladus marinus (Cienk.) Waern (class prasinophyceae)

Growth of Prasinocladus marinus (Cienk.) Waern was maximal in media of 35‰ salinity, showing a five-fold increase over a 7-day period. At lower salinities (10–27.5‰) growth varied between 55–90% and at 45‰ was 85% of that in ‘normal’ sea water. Yield of extracellular carbohydrate was higher than in ‘normal’ sea water in lowered salinities and at 45‰ over a 42-day period. Larger quantities of extracellular carbohydrate were obtained at all stages of growth in media of 15‰, compared with those of 35‰ and 45‰. Whilst growth forms were markedly different at low salinities, DNA measurements indicated that similar quantities of cell material were present. Mannitol and glucose were the principal carbohydrates identified in intracellular material, and glucose in the extracellular products. Chloroplast pigment synthesis appears unaffected by growth at 15‰, 35‰, and 45‰.     

Temperature and salinity effects on the respiratory metabolism of the first zoeal stage of Macrobrachium holthuisi Genofre & Lobão (decapoda: Palaemonidae)

Oxygen consumption rates of stage I Macrobrachium holthuisi Genofre & Lobão zoeae were measured in 24 different temperature and salinity combinations using Cartesian diver microrespirometers. Metabolic rates varied little with salinity at 15°C while at 20°C a marked elevation occurred in 0 and 35‰ At 25°C, a slight elevation occurred in 0‰; rates remained constant, however, in the other salinities. At 30°C, respiratory rates were similar to those recorded at 25°C except for decreases at 0 and 28‰ salinity. Q₁₀ values in the different salinities were usually highest between 15 and 20°C. Statistical analyses showed that while both temperature, salinity and their interaction significantly influenced larval respiratory rates, temperature had the more pronouced effect. Larval metabolism is salinity independent over the salinity range encountered in the larval biotope (7–21‰) at temperatures of 15–30°C.     

Environmental factors affecting hatching of rotifer (Brachionus plicatilis) resting eggs

Hatching experiments were carried out on a population of Brachionus plicatilis (Dor strain) resting eggs produced in batch laboratory cultures under controlled conditions and then stored for at least one month at 4 °C in the dark. Light was found to be obligatory for termination of dormancy. Over the temperature range of 10–30 °C (at 9.0‰ salinity), hatching was optimal (40–70%) at 10–15 °C and decreased linearly with the rise in incubation temperature. Resting eggs incubated over a salinity range of 9–40‰ (at 15 °C) showed optimal hatching at 16‰. Incubation of resting eggs in distilled water permitted normal embryonic development, but neonates died at eclosion. Presence of algae, Chlorella stigmatophora (0.5 × 10⁶ cell ml^?1), was found to aid hatching.     

Osmotic and ionic regulation in the western rock lobster Panulirus longipes (Milne-Edwards)

The western rock lobster, Panulirus longipes (Milne-Edwards) is poikilosmotic over its tolerated salinity range, 25–45 ‰. Blood sodium is accumulated while chloride concentration is reduced. Sodium and chloride vary directly with the external salinity, although maintaining their differences in the same proportions as at normal salinity (36.0 ‰). Calcium is accumulated, ranging from over 150% at salinity 20 ‰ to about 117% at salinity 45 ‰. Potassium concentration is equivalent to the external at normal salinity, but is increased with lowered and decreased with raised salinity. Magnesium is reduced to about one-third that of the external concentration over the salinity range 20–40 ‰, but regulation begins to break down at 45 ‰. Individual ions exhibit, therefore, a range of regulation types, from poikilosmotic to homoiosmotic.Equilibrium for sodium, chloride, and calcium is attained in 10 h at salinities of 25, 30, 40 and 45 ‰ respectively. Rate constants for this exchange are linearly related to salinity differential, and rapid osmotic adjustment is by high permeability, equal in both directions, probably mainly via the gills. Muscle appears to act as a salt pool for sodium, chloride, and potassium but not for magnesium and calcium. Salt-loading causes a slight salt diuresis, the salts being excreted, probably via the gills. Except for calcium, there is no excretion of salt into the gut, but there is evidence of an exchange of chloride with another anion. Magnesium excretion is slow, and in the absence of osmotic stress possibly occurs via the antennal glands. All the ions examined appear to be regulated independently.     

Respiratory and osmoregulatory responses of the hermit crab, Clibanarius vittatus (Bosc), to salinity changes

Intertidal hermit crabs were stepwise acclimated to 10, 20, and 30‰ salinity (S) and 21 ± 1 °C. Hemolymph osmolality, sodium, chloride, and magnesium were isosmotic (isoionic) to ambient sea water at 30‰ and hyperosmotic (hyperionic) at 20 and 10‰ S, while hemolymph potassium was significantly hyperionic in all acclimation salinities. Total body water did not differ significantly at any acclimation salinity. Oxygen uptake rates were higher in summer-than winter-adapted crabs. No salinity effect on oxygen consumption occurred in winter-adapted individuals. Summer-adapted, 30‰ acclimated crabs had a significantly lower oxygen consumption rate than those acclimated 10 and 20‰ S. Crabs exposed to 30 10 30‰ and 10 30 10‰ semidiurnal (12 h) and diurnal (24.8 h) fluctuating salinity regimes showed variable osmoregulatory and respiratory responses. Hemolymph osmolality followed the osmolality of the fluctuating ambient sea water in all cases, but was regulated hyperosmotically. Hemolymph sodium, chloride, and magnesium concentrations were similar to hemolymph osmolality changes. Sodium levels fluctuated the least. Hemolymph potassium was regulated hyperionically during all fluctuation patters, but corresponded to sea water potassium only under diurnal conditions. The osmoregulatory ability of Clibanarius vittatus (Bosc) resembles that reported for several euryhaline brachyuran species. The time course of normalized oxygen consumption rate changed inversely with salinity under semidiurnal and diurnal 10 30 10‰ S fluctuations. Patterns of 30 10 30‰ S cycles had no effect on oxygen consumption rate time course changes. The average hourly oxygen consumption rates during both semidiurnal fluctuations were significantly lower than respective control rates, but no statistical difference was observed under diurnal conditions.     

Effect of silymarin on plant tissue cultures

The effect of silymarin complex on various types of expiants differing in their nutrition requirements was investigated. The growth of tumorous periwinkle (Catharanthus roseus [L.] G. Don) callus tissue was still identical with the control tissue at the silymarin concentration of 35 mg in 1000 ml of the nutrient medium. However, this silymarin concentration totally inhibited the growth of habituated periwinkle callus tissue; in the presence of 10 mg of silymarin, the growth of this tissue was similar to that of the corresponding tissue grown without silymarin. The growth of tobacco callus tissue (D-strain) requiring for its growth kinetin was reduced by 46.2% at the concentration 10 mg of silymarin in 1000 ml of nutrient medium, but its dry weight was increased by 21% in comparison with the control. Silymarin was most effective on the growth of callus derived from tobacco (Nicotiana glauca Grah:) stem pieces; callogenesis was observed in control tissue in 89.5% cases while in the presence of silymarin (10 mg) only in 48.6%. The primary callus growth was strongly inhibited, too (by 89.9%). The organogenesis onset was never observed on tobacco stem pieces cultured on a nutrient medium with kinetin and IAA in the presence of silymarin. When all types of expiants were transferred from the medium with silymarin on control medium, normal growth appeared very soon and the differences between the experimental and control expiants were smoothed out during two months. These results indicate that the observed changes might be due to the blocking of membrane system permeability leading to an insufficient supply of cells with nutrients and growth substances.     

Changes in plasma osmolality and Na+/K+ ATPase activity of juvenile obscure puffer Takifugu obscurus following salinity challenge

The osmoregulatory capabilities of 6-month-old juvenile obscure puffer Takifugu obscurus, transferred directly from fresh water to different salinities (0‰, freshwater control; 10‰; 20‰ and 30‰), were studied over an 8-day period. After transfer, plasma osmolality of the fish at 30‰ was significantly higher than those at all other salinities throughout the experiment. The Na⁺/K⁺ ATPase activity in the gills of the fish treated with various salinities increased significantly, peaking at 48 h, then decreased gradually to the control level at 192 h. Similar fluctuation trends of the Na⁺/K⁺ ATPase activity were observed in the kidneys. Modified Gaussian model provided accurate fits for the time-course changes in the Na⁺/K⁺ ATPase activities after abrupt salinity challenge. The results demonstrated that obscure puffer has strong capacity to tolerate abrupt salinity changes and can osmoregulate well over a wide range of salinities even in juvenile stage.     

Responses of Embryos and Larvae of the Starfish <Emphasis Type="Italic">Asterias amurensis</Emphasis> to Changes in Temperature and Salinity

We studied the effects of different combinations of temperature (5, 10, 14, 17, 20, and 22°C) and salinity (from 32 to 8‰) on the development of the starfish Asterias amurensis Lutken from Vostok Bay, Sea of Japan. Embryonic development is the most vulnerable stage; it passes successfully at 10–17°C and the salinity range of 32 to 26‰. Blastulae are the most tolerant of changing environmental factors. They survive and develop at the temperatures of 5–17°C and in the salinity range of 32–18‰. Gastrulae and bipinnariae survive under higher temperature values and salinity from 32 to 20‰. The tolerance for decreased salinity during the process of fertilization and in the latest stage of development, the brachiolaria with the developing juvenile starfish, was confined to the salinity range of 32–22‰, which agrees with the tolerance of adult starfish Asterias amurensis. Thus, for normal development of the Amur starfish in the early stages, some particular conditions of temperature and salinity are required. This is, probably, due to adaptive capabilities of each developmental stage and the peculiarities of the ecological conditions at particular depths.     

Salinitätstoleranz des parasitischen CiliatenIchthyophthirius multifiliis

Eels,Anguilla anguilla (L.), infested withIchthyophthirius multifiliis Fouquet, were exposed to different salinity levels (8, 15, and 30 ‰). Following exposure to 15° C for 2 to 4 weeks the parasite was able to survive on 20 cm long, young eels. This demonstrates thatI. multifiliis is not being damaged below the epidermis of its host; in form of free cysts or as swarmers, it is, however, no longer able to cause infection. In a second experiment, 2 eels of 40 cm length were infested withI. multifiliis at 15° C in 8 ‰ S; however, in 15 ‰ S, the parasite disappeared after 4 weeks.     

The combined effects of temperature,salinity, and declining oxygen tension on oxygen consumption in the marine pulmonate Amphibola crenata (Gmelin, 1791)

Oxygen consumption of Amphibola crenata (Gmelin) was measured in various salinity-temperature combinations (< 0.1‰ to 41‰ salinity and 5 to 30°C) in air, and following exposure to declining oxygen tensions. In all experimental conditions, respiration varied with the 0.44 power of the body weight (sd = 0.14). The aquatic rate was consistently higher than the aerial rate of oxygen consumption, although at 30 °C the two rates were similar. Oxygen consumption increased with temperature up to 25 °C in all salinities; the lowest values were recorded at temperatures below 10 °C and at 30 °C in the most dilute medium. At all exposure temperatures, the oxygen consumption of Amphibola decreased regularly with salinity down to 0.1 ‰, and following exposure to concentrated sea water (41‰). Salinity had the least effect at 15 °C which was the acclimation temperature. In general, all of the temperature coefficients (Q₁₀ values) were low, < 1.65. However, Q₁₀ values above 2.8 were recorded at a salinity of 17.8‰ between 10 and 15 °C. Oxygen consumption of all size classes of Amphibola was more temperature dependent in air than in water and small individuals show a greater difference between their aerial and aquatic rates than larger snails. The rates of oxygen consumption in declining oxygen tensions were expressed as fractions of the rates in air saturated sea water at each experimental salinity-temperature combination. The quadratic coefficient B₂ becomes increasingly more negative with both decreasing salinity and temperatures up to 20 °C. At higher temperatures (25 and 30 °C) the response is reversed such that O₂ uptake in snails becomes increasingly independent of declining oxygen tensions at higher salinities. On exposure to a salinity of 4‰, Amphibola showed no systematic response to declining oxygen tension with respect to temperature. The ability of Amphibola to maintain its rate of oxygen consumption in a wide range of environmental conditions is discussed in relation to its potential for invading terrestrial habitats and its widespread distribution on New Zealand's intertidal mudflats.     

EFFECT OF INITIAL SIZE ON GROWTH OF PLANT TISSUE CULTURES

Caplin , Samuel M. (LOS Angeles State Coll., Los Angeles, Calif.) Effect of initial size on growth of plant tissue cultures . Amer. Jour. Bot. 50(1): 91–94. Illus. 1963.—Using different slice thicknesses and cannula diameters, cylindrical expiants of different size were obtained from tubers of Jerusalem artichoke. Slice thickness ranged from 0.5 to 2.5 mm and expiant diameter from 1.30 to 3.22 mm. Initial fresh weights ranged from 2.3 to 29.8 mg, a 13-fold range. After 17 days growth at 26 C in 10 ml basal medium containing 15% coconut milk, relative increase was inversely related to initial weight and ranged from 36-fold for 2.3-mg expiants to 8-fold for 29.8-mg expiants, a range of 4.5 × in relative increase between the largest and smallest cultures. Secondary phloem expiants of the same diameter from carrot root slices of different thickness, all removed at 1 mm from the cambium, also showed decreased relative growth with increasing size of explant; this was true for each of the different diameters used. Further investigation showed clearly that the growth of the large inocula was not limited by the amount of culture medium used during the period of culture. Initial weight has, therefore, a decided effect on the rate of relative growth.     

Formation of adventitious buds in needle primordia isolated from embryonic shoots of adult picea abies

Needle primordia were isolated from embryonic shoot expiants of 88 year oldPicea abies (Norway spruce) in order to develop a method for micropropagation using adult mother trees. The isolation of needle primordia in combination with frequent subculture, stimulated the formation of peripheral meristematic tissue in expiants cultured on a modified Murashige & Skoog medium supplemented with cytokinin. Transfer of isolated needle primordia with peripheral meristematic tissue to medium without growth regulators, and with a reduced ratio of NH+4/ NO?3, resulted in the formation of adventitious buds.     

Anatomical and physiological effects of silver thiosulfate on ethylene-induced abscission inColeus

Petioles of expiants ofColeus blumei Benth. exposed to 20 μl/l ethylene abscised within 36 h. Pretreatment of expiants with 4 mM silver thiosulfate (STS) inhibited ethylene-induced abscission. Delaying treatment with STS reduced its effectiveness in retarding ethylene-promoted abscission, suggesting that some events leading to abscission are initiated during the first hours of ethylene treatment. Microscopic study of abscission zones of ethylene-treated expiants showed greatly increased amounts of rough endoplasmic reticulum, disruptions of the plasma membrane, and some cell separation in the region of the middle lamella. Pretreatment with STS prevented ethylene-induced reorganization of the endomembrane system and the subsequent middle lamellar dissolution.     

Acetylcholinesterase activity of the polychaete Nereis diversicolor: effects of temperature and salinity

In order to estimate the potential use of the mean wholebody acetylcholinesterase (AChE) activity from the ragworm Nereis diversicolor for the biological assessment of pollution by anticholinesterase agents in estuarine areas, we measured the effects of the main abiotic factors (i.e. temperature and salinity) on AChE activity. We report here that AChE activity tends to decrease in individuals sampled in tanks at a salinity of 30‰ as temperature increases. No tendencies in the evolution of AChE activity were observed in individuals sampled in tanks at a salinity of 15‰. In contrast, salinity seems to have a greater effect on AChE activity than temperature. At a temperature of 12°C, a salinity of 30‰ provokes a significant transient increase of AChE 2 days after the beginning of the maintenance period compared with a salinity of 15‰. The effects are short-term stress effects. We noticed only a transient increase of AChE activity between 2 days for individuals maintained in tanks at temperature of 20°C and salinity of 15 and 30‰, respectively, and 8 days for individuals maintained in tanks at salinity of 30‰ and at a temperature of 12°C after the beginning of the maintenance period, confirming the more pronounced effect of salinity over temperature.     

Innate immunity of the white shrimp Litopenaeus vannamei weakened by the combination of a Vibrio alginolyticus injection and low-salinity stress

White shrimp Litopenaeus vannamei were reared at a salinity of 35‰ without a Vibrio alginolyticus injection (unchallenged group), and other shrimp were reared at 35‰, injected with tryptic-soy broth (TSB)-grown V. alginolyticus at 1.8 × 10⁵ colony-forming units (cfu) shrimp^?1 (challenged group), and then examined for the hyaline cell (HC) count, granular cell (GC, including semi-granular cell) count, total haemocyte count (THC), phenoloxidase (PO) activity, respiratory burst (RB) and superoxide dismutase (SOD) activity after transfer to 35‰ (control), 25‰, 20‰, and 15‰ for 1, 6, 12, 24, 72, and 120 h. Results indicated that the haemocyte count, PO activity, RB, and SOD activity of unchallenged shrimp and challenged shrimp that were transferred to low-salinity levels all began to significantly decrease at 6, 6, 6, and 1 h, respectively, and reached the lowest levels at 12 h. HC, GC, the THC, PO activity, RB, and SOD activity of unchallenged shrimp that were transferred to 15‰ decreased by 53%, 41%, 49%, 68%, 39%, and 62%, whereas those parameters of challenged shrimp that were transferred to 15‰ decreased by 79%, 78%, 79%, 82%, 54%, and 72%, respectively after 12 h compared to control shrimp. These immune parameters began to recover after 24–72 h for both unchallenged shrimp and challenged shrimp. We concluded that the innate immunity was weakened in white shrimp L. vannamei that received combined stresses of a V. alginolyticus injection, and low-salinity transfer. It was also concluded that shrimp with respectively 21%, 18%, 46%, and 28% lower THC, PO activity, RB, and SOD activity of the original values would be killed due to decreases in their immunity, and resistance to V. alginolyticus infection. Shrimp farming should be maintained at a constant high salinity level to prevent exacerbated decreases in innate immune parameters of shrimp when infected by a pathogen coupled with low-salinity stress leading to mortality.     

Effects of salinity on the respiration and heart rate of the common mussel, Mytilus edulis L., and the black chiton, Katherina tunicata (Wood)

The rate of oxygen consumption of stepwise acclimated Mytilus edulis L. increased linearly from 30 to 10‰ salinity (S) while that of Katherina tunicata (Wood) was not significantly different between 10 and 30‰ S. Heart rate was 21–22 and 17–18 beats m^?1 in Mytilus edulis and Katherina tunicata, respectively, and no difference was found in the heart rate of either species acclimated stepwise to 10, 20 or 30‰ S. The average oxygen consumption rate of Mytilus edulis exposed to 12 h, 30-10-30 and 10-30-10‰ S cycles of fluctuating salinity was significantly lower than the respective control rate: there was a similar response during the 30-10-30‰ S cycle in Katherina tunicata. The respiration rate of Mytilus edulis and Katherina tunicata declined as salinity deviated from the control salinity and increased as salinity returned to the control salinity. The rate of oxygen consumption by K. tunicata varied directly with the ambient salinity during the 10-30-10‰ S cycle. The average heart rate of Mytilus edulis was significantly lower during cyclic changes in salinity than at the respective control salinities; a similar relationship existed for Katherina tunicata during the 10-30-10‰ S cycle. Heart rate of Mytilus edulis varied in a parallel manner with oxygen consumption during both cycles. Katherina tunicata heart rate was relatively constant and could not be fitted to a regression line during the 10-30-10‰ S cycle. The normalized heart rate increased to 113% of control at 10‰ S of the 30-10-30‰ S cycle and returned to the control rate by 12 h. The oxygen consumption and heart rate of these two species are not directly coupled to regulation of water volume because different responses are observed with respect to salinity although there is poor water volume regulation in both species.     

Salinity influence on the ecology of phytoflagellate blooms in lower new york bay and adjacent waters

The possible rôle of salinity in phytoflagellate blooms in New York Harbor estuarine and oceanic waters was examined by culture studies of the dominant species. Massartia rotundata (Lohmann) Schiller (=Katodinium rotundatum (Lohmann) Loeblich III) grew best in the range 24–30‰, Olisthodiscus luteus Carter in the range 10–36‰, and Prorocentrum micans Ehrenberg in the range 27–36‰. The optimum ranges for all three species sufficiently match the characteristic bay-ocean range (17–32‰) to eliminate salinity tolerance as a major factor in bloom development. Laboratory simulation of the change from brackish-river to ocean water salinities, by culture preconditioning at 20‰ or at 30‰ before inoculation into a salinity gradient, shows stress effects on growth, especially in Massartia rotundata and Olisthodiscus luteus. It is speculated that salinity stress may temporarily affect the development of particular blooms of these two species.     

Adaptive responses of embryos and larvae of the heart-shaped sea urchin <Emphasis Type="Italic">Echinocardium cordatum</Emphasis> to temperature and salinity changes

The combined effects of temperature (8, 12, 14, 17, 20, 22 and 25°C) and a salinity decrease from 36 to 12‰ on the development of the sea urchin Echinocardium cordatum (Pennant) were studied. Embryonic development proved to be the process most vulnerable to a salinity decrease. It was completed successfully at 8–20°C within a narrow salinity range of 36–28‰ Larvae at the most resistant stage, the blastula, survived at 12–22°C and a salinity of 36–18‰. Larvae at the most sensitive stage, pluteus I with the first pair of arms, died even in a favorable environment, a temperature of 17–20°C and a salinity of 34–28‰. That may be related to qualitative alterations during skeleton formation and to transition to phytoplankton feeding. The resistance of larvae to variations in environmental factors gradually increased in the pluteus II and III stages; however, it significantly decreased before the settling of the larvae. Larvae that were 37 days old survived at a temperature of 14–20°C within a salinity range of 36–22‰ and at 22 and 25°C, they survived at a salinity of 36–24‰; however, all the larvae became abnormal at 25°C. The larvae settled earlier on sand inhabited by adult individuals of E. cordatum than on sand from other locations, and they settled faster at 20–25°C, than at 14 and 17°C. The juveniles, if lacking an opportunity to burrow in the sand, died within 14 days after settling.     

The effect of salinity on coelomic fluid osmolyte concentration and intracellular water content in Luidia clathrata (Say) (Echinodermata: Asteroidea)

• 1.1. The concentrations (mM) of osmolytes in the coelomic fluid of Luidia clathrata kept at 25‰S seawater (control individuals) were: 345, Na⁺; 10, K⁺; 10, Ca²⁺; 44, Mg²⁺; 387, Cl⁻; 0.67, amino acids; 0.09, NH₄⁺.
• 2.2. When individuals were transferred from 25‰S to 15‰S or 35‰S, the concentrations of inorganic ions in the coelomic fluid usually equilibrated within 24hr and became the same as those in the medium.
• 3.3. The intracellular water content (g intracellular H₂O/g solute-free dry tissue) of the pyloric caeca and tube feet of control individuals throughout the experiment was 2.13 and 5.40, respectively.
• 4.4. In tissues of individuals transferred to 15‰S, the intracellular water content increased by an average 50% in 12 hr but returned to 19% above control levels during 1 week.
• 5.5. In tissues of individuals transferred to 35‰S, the intracellular water content decreased by an average 17% in 12 hr and did not change during 1 week.
• 6.6. Luidia clathrata is an osmoconformer and partial cell volume regulator within the seasonal salinity range it encounters.

     

Cellulase activity and localization during induced abscission of Coleus blumei

Treatment with dimethipin (2,3-dihydro-5,6-dimethyl-1,4-dithiin 1,1,4,4 tetroxide) inhibited the increase in cellulase activity and decrease in breakstrength associated with the normal course of abscission in Coleus. Application of the surfactant UBI-1126 (Emery OAL 20 in isopropyl alcohol) increased cellulase activity and accelerated the process of abscission in Coleus expiants within 24 h of application. Cellulase activity was localized histochemically at the electron microscopic level in surfactant-treated tissue. The enzyme activity was localized primarily in the cell wall, middle lamella, and paramural bodies of abscission zone cells.