Induction of tolerance to fast desiccation in black spruce (Picea mariana) somatic embryos: relationship between partial water loss,sugars, and dehydrins

Events associated with the induction of tolerance to fast desiccation in black spruce ( Picea mariana ) somatic embryos were investigated. An experimental approach using an initial period of partial water loss was developed to induce either no, partial, or complete tolerance to fast desiccation. Tolerance to subsequent fast desiccation was not promoted by decreasing embryo water content from 1.5 to 1.1 g H₂O g⁻¹ DW (g g⁻¹) throughout the first 24 h of slow desiccation. However, tolerance increased from 10 to 95% germination during the second 24-h period of slow desiccation after partial water loss from 1 to 0.55 g g⁻¹. Emphasis was also placed on the relationship between observed tolerance, and sugar and dehydrin contents. Compared to controls, sucrose content in embryos doubled after 24 h of slow desiccation and more than tripled after 48 h. Conversely, starch content was decreased by one half after 24 h and by three quarters after 48 h. Sucrose abundance and raffinose occurrence after 48 h of slow desiccation were congruent with complete tolerance to fast desiccation. The period of slow desiccation between 24 and 48 h also increased the content of a 24-kDa dehydrin and the appearance of a 42-kDa dehydrin. The relationship between partial water loss, sugars and dehydrins is discussed with respect to tolerance to fast desiccation in black spruce somatic embryos.     

Physiological adaptation during cell dehydration and rewetting of a newly-isolated Chlorella species

A new terrestrial species of green alga ( Chlorella sp. strain DT) that survives under extremely dry conditions was isolated from an arid mountainous area of Taiwan. The water content of the cells dropped to less than 3% after 3 h dehydration at 42°C. The dried cells could regrow if they were rewetted. The photosynthetic activity of the cells ceased following 2 h of dehydration, but respiratory activity was still detectable after 3 h desiccation. During the rewetting process, respiration increased immediately and then decreased to a steady level after 5 days of rewetting, matching the net photosynthetic oxygen evolution. The cells had 38% neoxanthin (1520 nmol g⁻¹ dry weight), 39% lutein (1580 nmol g⁻¹ dry weight), and 14% violaxanthin (570 nmol g⁻¹ dry weight) of total carotenoids, but only 5.1%β-carotene (210 nmol g⁻¹ dry weight), 3.8%α-carotene (150 nmol⁻¹ g dry weight) and a trace of antheraxanthin and zeaxanthin. Upon dehydration, zeaxanthin and carotene contents increased and recovered to normal levels after 8 days of rewetting. The photoprotective xanthophyll cycle was found in these cells. It appears that the energy dissipation process for preventing photodamage is perhaps one of the tolerance mechanisms in this Chlorella strain.     

Hydration, protons and onset of physiological activities in maize seeds

Dry maize ( Zea mays L.) seed components, namely, embryo and endosperm, provide model materials for studies on water-dependent mechanisms in cellular function. We explored the thermodynamics of hydration for both tissues, along with their dielectric behavior, as a function of water content. In addition, we evaluated the direct current (DC) conductivity due to water protons. Our data on embryo tissue show large enthalpic and entropic peaks at water content [h, in g H₂O (g dry sampie)⁻¹] around 0.08 g g⁻¹, indicating very tight binding and ordering of water molecules. With increasing water content both enthalpy and entropy decrease, and the completion of primary hydration requires h ∼ 0.26 g g⁻¹. Data for endosperm tissue show the absence of such an enthalpic peak and a reduced degree of ordering for h < 0.10 g g⁻¹. The DC protonic conductivity shows explosive growth above a threshold hydration level h_c= 0.082 g g⁻¹ and h_c= 0.12 g g⁻¹, for embryo and endosperm, respectively. Protonic conduction can be considered within the framework of a percolation modell characterized by a hydration threshold and by a power law increase in conductivity with further hydration. The critical exponent of the power law is in agreement with theory for a two-dimensional percolative process. This percolative water-assisted behavior reflects the presence of an extended network of water molecules adsorbed on the surface of proteins and/or membranes inside cells. We consider this percolative protonic conduction as being a prerequisite to respiration processes.     

Diet selection and energy and water budgets of the common spiny mouse Acomys cahivinus

The common spiny mouse, Acomys cahirinus (body mass=47 g), is widely distributed in Israeli deserts where it inhabits natural crevices on rocky slopes. This omnivorous rodent consumes a varied diet, and in particular snails. We determined diet selection and energy and water balances of spiny mice when they were offered snails and seeds. The spiny mice maintained steady state body mass. Dry matter consumption of snails was 0.014 g* g⁻¹.-d⁻¹ and of seeds was 0.049g*.g⁻¹d⁻¹ for a total of 0.063 g*g⁻¹*d⁻¹. Total water intake was 0.101ml-g⁻¹.d⁻¹ and metabolizable energy intakewas 0.990 kJ. gxs^-1.d⁻¹ for a ratio (ml: kJ) of 0.102. This ratio was similar to that reported in a previously published study on free-living spiny mice. We concluded that snails and seeds allowed spiny mice to fulfil their energy and water requirements with minimal dry matter and fresh matter intakes. Furthermore, spiny mice selected a diet that provided them with a water (ml) to energy (kJ) ratio of approximately 0.1, although it appeared that they are able to survive on a much drier diet.     

Acquisition of desiccation tolerance in soybeans

The entry into a desiccation-tolerant state is a major developmental component of seed maturation. Development of desiccation tolerance of embryonic axes of soybean [Glycine max (L.) Merrill cv. Chippewa 64] was studied by measuring changes in electrolyte leakage. germination and relative growth rate after axes were rapidly air-dried to various water contents. Axes acquired the full capacity for germination at 34 days after flowering (DAF). and reached physiological maturity (maximum dry weight) at 48 DAF. When dried to water content h = 0. 08 (g water g⁻¹ dry weight). few axes germinated before 42 DAF. but more than 90% germinated after 48 DAF. However, electrolyte leakage of rehydrated axes showed a linear decline from 30 to 55 DAF. For developing axes there was a critical water content or desiccation threshold. which could be estimated by using the electrolyte leakage method. The threshold of desiccation tolerance decreased gradually from h = 1. 10 to 0. 18 as axes matured from 28 to 55 DAF. The development of desiccation tolerance continued after physiological maturity at 48 DAF. We conclude that the acquisition of desiccation tolerance of soybean axes is a gradual event, rather than an abrupt transition.     

Storage behavior of Typha latifolia pollen at low water contents: Interpretation on the basis of water activity and glass concepts

Germplasm must be stored under optimal conditions to maximize longevity and efficiently maintain genetic resources. In order to identify optimal storage conditions, we investigated the effects of temperature (−5 to 45°C) and water content (<0.17 g H₂O g⁻¹ dry weight) on longevity of Typha latifolia L. pollen. Longevity was highest at water contents corresponding to storage relative humidity (RH) of 11‐15% which corresponded to the shoulder of water sorption isotherms. Also coinciding with this shoulder were abrupt changes in heat capacity of water present in the pollen. Consistent with changes in isotherms with temperature and the concept of critical RH for storage, optimum water contents increased with decreasing temperature. An attempt was made to explain the aging behavior according to the glass concept. The water content‐temperature combinations of optimal storage were found to be below the glass transition curve, indicating that optimum storage conditions are achieved when intracellular glasses are present. We also found a change in activation energy of aging in Arrhenius plots around T_g, demonstrating a change in aging kinetics when the glassy state is lost. We concluL that T_g curves cannot be used solely to predict precise conditions of optimum storage, but might be useful for predictions of storage longevity above optimum water contents. The data imply that too much drying reduces longevity and should be avoided, particularly when cryogenic storage is considered.     

Carbon, nitrogen and caloric content of eggs, larvae, and juveniles of the walleye pollock, Theragra chalcogramma

Measurements of dry weight (wt), carbon (C), nitrogen (N) and calories were made on walleye pollock eggs (0.24 mg, 35.3% C, 8.3% N, and 4.6 kcal g⁻¹ dry wt), larvae (0.16 g, 42.9% C, 11.1% N and 5.1 kcal g⁻¹ dry wt) and juveniles (22.4 g, 47.2% C, 9.0% N and 5.6 kcal g⁻¹ dry wt). For juvenile fish (9–360 g wet wt) the measured values were related to dry weight and Fulton's condition factor index (CFI) by regression models. The CFI was a better predictor of body composition than dry weight. As CFI improved from a minimum starvation level of 0.42 to a maximum of 1.16, body caloric content, percentage C, and the C/N ratio increased (kcal g⁻¹ dry wt = 4.4 CFI + 1.7, percentage carbon = 49.7 CFI^0.5, C/N ratio = 5.0 CFI + 0.9), while percentage N and percentage ash decreased (percentage N =−3.5 CFI + 12.1; percentage ash = 9.1 CFI^−1.4). The results of this study suggest that seasonal C, N and caloric content of young pollock can be estimated from measurements of Fulton's condition factor index.     

Seed dimorphism in Salicornia europaea: Nutrient reserves

Median and lateral seeds of Salicornia europaea L. were separately analysed for their sizes and nutrient reserves. The mean air-dry weight of a single median and lateral seed was 0.31 and 0.25 mg, respectively. The composition as well as the concentration of the nutrient reserves were similar in both seed types. The bulk of the cations was derived from K⁺, followed by Mg²⁺, Na⁺ and Ca²⁺. The chloride content was somewhat higher than the sodium content, and phosphate was equalled by acid soluble Ca²⁺ and Mg²⁺. Starchy compounds and sucrose were present in equal amounts, each of them accounted for about 50% of the carbohydrates. Glucose and fructose were less than 1%. Protein-nitrogen (ethanol-insoluble N) was about 34 g (kg dry seeds)⁻¹. About 7 g (kg dry seeds)⁻¹ was ethanol-soluble nitrogen, of which 10% was derived from amino acids. The total lipid content was more than 290 g (kg dry seeds)⁻¹, 65% were calculated to be glycerides. More than 90% of the fatty acids consisted of linoleic and oleic acids, the majority (72%) of which was linoleic acid.     

Tolerance of coffee (Coffea spp.) seeds to ultra-low temperature exposure in relation to calorimetric properties of tissue water, lipid composition, and cooling procedure

The effect of exposure to ultra-low temperature (liquid nitrogen, LN) on viability of seeds desiccated to various water contents was investigated in 9 coffee species. Three groups of species could be distinguished based on seed survival after LN exposure. In group 1 species, no seedling production could be obtained after LN exposure due to endosperm injury. In group 2 species, recovery was very low or nil after rapid cooling, and only moderate after slow cooling. In group 3 species, very high percentages of seedling development were observed after both rapid and slow cooling. A high interspecific variability for the high moisture freezing limit was observed within the species of groups 2 and 3, since it ranged from 0.14 to 0.26 g H₂O g⁻¹ dry weight. A very highly significant correlation was found for those species between the unfreezable water content, as determined from DSC analysis, and the high moisture freezing limit of their seeds. No significant correlation was found between seed lipid content, which varied from 9.8 to 34.6% dry weight, and survival after LN exposure. However, a negative relationship was found between seed unfreezable water content and lipid content. Interspecific differences in fatty acid composition of seed lipids resulted in a high variability in the percentage of unsaturated fatty acids, which ranged from 28.7 to 54.4% among the 9 species studied. For all species studied, a highly significant correlation was found between the percentage of unsaturated fatty acids and the percentage of seedling recovery after rapid or slow cooling.     

Nutrient reserves of Salicornia europaea seeds

Seeds of Salicornia europaea L. were analyzed for their nutrient reserves. The content of potassium and sodium was 216 and 39 mmol (kg dry seeds)^-1, respectively. Calcium and magnesium accounted for 30 and 138 mmol (kg dry seeds)^-1, respectively. Whereas most of the alkali metals were water soluble, the alkaline earth metals were mostly acid soluble. The acid-soluble calcium plus magnesium corresponded well with the acid-soluble phosphate. Chloride was accumulated to a level equivalent to that of sodium. Carbonate was present at a concentration of 9 mmol (kg dry seeds)^-1. Carbohydrates accounted for 93 g (kg dry seeds)^-1, nearly half of which was derived from sucrose. Fructose and glucose were present only in traces. Total nitrogen was determined to be 55 g (kg dry seeds)^-1, 16% of which was diethylether soluble. The remaining nitrogen was separated into 39 g (kg dry seeds)^-1 ethanol-insoluble and 8 g (kg dry seeds)^-1 ethanol-soluble nitrogen. About 10% of the ethanol-soluble nitrogen were derived from amino acids. Total lipid content was about 280 g (kg dry seeds)^-1. The alcoholic component of the storage lipids was glycerol and the glycerides were calculated from gas chromatography to be 66% of the total lipids. About 90% of the fatty acids consisted of unsaturated acids, linoleic and oleic acid, the majority (77%) of which was linoleic acid.     

The use of body water, sodium, potassium and calcium content to investigate the nutritional status of first year Atlantic salmon parr in two Scottish Highland streams

Water content of Atlantic salmon parr fell from about 84% at emergence (late May) to just under 79% in September but rose again towards March. Na⁺ content consequently rose from 3·3 mg g⁻¹ dry wt at the beginning of June to 6·2 mg g⁻¹ in early July. It then fell to 4·4 mg g⁻¹ in September, rising again towards March. K⁺ content rose to a maximum in July to stabilize at 16·6 mg g⁻¹ dry wt in September. The resultant Na⁺/K⁺ ratio peaked at 0·43: 1 in mid-June, falling to a minimum in mid-August but rising again in March reflecting changes in the relative proportions of intra and extracellular water. The changes in whole-body chemistry suggest a period of nutritional stress immediately after emergence and during the winter. In streams at higher altitude and of lower nutrient status, nutritional stress during the winter appears to be more severe.     

Estimation of the bioenergetic costs of fruit and other organ synthesis in apple

The specific bioenergetic cost of apple fruit ( Malus domestica Borkh. cv. Braeburn) growth was calculated from seasonal elemental analysis (for carbon, hydrogen, nitrogen, and sulphur) and ash data. Specific cost changed during fruit development and at harvest was 1.16 g glucose g⁻¹ dry weight of which 0.142 g glucose g⁻¹ dry weight was consumed in growth respiration. Comparisons of those end-of-season values with those from a range of other apple cultivars (early through late maturing, and with a range of sugar/acid ratios) showed little difference, despite variations in elemental composition. Specific costs ranged between 1.15 and 1.16 g glucose g⁻¹ dry weight, and growth respiration between 0.136 and 0.148 g glucose g⁻¹ dry weight. Specific costs were also calculated for leaves (extension and spur), wood (1 and 2 year), and trunk and roots (fine and coarse). Leaves had the greatest cost (mean 1.44 g glucose g⁻¹ dry weight), then wood and trunk (mean 1.38 g glucose g⁻¹ dry weight), and fruit had the smallest. Specific growth costs were also calculated from heats of combustion data. Little difference was observed between the two methods, but the values calculated from the elemental analysis data tended to be higher than those calculated from the heats of combustion data.     

Content of low-molecular-weight thiols during the imbibition of Pea seeds

The metabolism of low-molecular-weight thiols was investigated in seeds of Pisum sativum L. cv. Kleine Rheinländerin during imbibition in water for 14 h. The amount of oxidized glutathione (GSSG) decreased from 319 nmol (g dry weight)⁻¹ in dry seeds to 38 nmol (g dry weight)⁻¹ within the first 14 h of imbibition. The decrease may have been due to the reduction of GSSG to reduced glutathione (GSH), catalyzed by the enzyme glutathione reductase (GR; EC 1.6.4.2). The enzyme activity was high in dry seeds [25 nkat (g dry weight)⁻¹] and decreased to 20 nkat (g dry weight)⁻¹ within 14 h of imbibition. The activity of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) decreased from 100 nkat (g dry weight)⁻¹ in dry seeds to 67 nkat (g dry weight)⁻¹ after 14 h of imbibition. Within 14 h the amount of γ-glutamyl-cysteine (γ-GC) decreased from 135 to 38 nmol (g dry weight)⁻¹, whereas the cysteine content rose from 81 nmol (g dry weight)⁻¹ in dry seeds to a maximum of 170 nmol (g dry weight)⁻¹ after 12 h of imbibition, which may be due to the degradation of γ-GC into cysteine.     

Energy density of anchovy Engraulis encrasicolus in the Bay of Biscay

The energy density ( E _D) of anchovy Engraulis encrasicolus in the Bay of Biscay was determined by direct calorimetry and its evolution with size, age and season was investigated. The water content and energy density varied seasonally following opposite trends. The E _D g⁻¹ of wet mass ( M _W) was highest at the end of the feeding season (autumn: c . 8 kJ g⁻¹ M _W) and lowest in late winter ( c . 6 kJ g⁻¹ M _W). In winter, the fish lost mass, which was partially replaced by water, and the energy density decreased. These variations in water content and organic matter content may have implications on the buoyancy of the fish. The water content was the major driver of the energy density variations for a M _W basis. A significant linear relationship was established between E _D g⁻¹ ( y ) and the per cent dry mass ( M _D; x ): y =−4·937 + 0·411 x . In the light of the current literature, this relationship seemed to be not only species specific but also ecosystem specific. Calibration and validation of fish bioenergetics models require energy content measurements on fish samples collected at sea. The present study provides a first reference for the energetics of E. encrasicolus in the Bay of Biscay.     

Survival of κ-carrageenan-encapsulated and unencapsulated Pseudomonas aeruginosa UG2Lr cells in forest soil monitored by polymerase chain reaction and spread plating

Abstract A method was developed for direct extraction, purification and amplification of DNA from forest soil. Eighty-two % of the DNA in Pseudomonas aeruginosa UG2Lr introduced into soil was recovered. The detection limit for the strain was approximately 800 cfu g⁻¹ of dry soil based on the polymerase chain reaction (PCR). Survival of κ-carrageenan-encapsulated and unencapsulated UG2Lr was monitored by antibiotic selective and bioluminescence-based nonselective plating and PCR-amplification of a tnsA fragment. After freeze-thaw treatment of soil samples, the unencapsulated UG2Lr declined from an initial population density of 1 × 10⁹ cfu g⁻¹ of dry soil to below the detection threshold of both selective (14 cfu g⁻¹ of dry soil) and nonselective (1 × 10³ cfu g⁻¹ of dry soil) plating. However, presence of nonculturable UG2Lr cells in the soil was revealed by PCR and resuscitation of the bacteria. Population density of the encapsulated UG2Lr increased from 2.7 × 10⁶ to 2.9 × 10⁸ cfu g⁻¹ of dry soil after a 3-week incubation at 22°C and declined to 6.3 × 10⁶ cfu g⁻¹ of dry soil after the freeze-thaw treatment.     

Triacylglycerol phase and ‘intermediate’ seed storage physiology: a study of Cuphea carthagenensis

Seeds with ‘intermediate’ storage physiology store poorly under cold and dry conditions. We tested whether the poor shelf life can be attributed to triacylglycerol phase changes using Cuphea carthagenensis (Jacq.) seeds. Viability remained high when seeds were stored at 25°C, but was lost quickly when seeds were stored at 5°C. Deterioration was fastest in seeds with high (≥0.10 g g⁻¹) and low (0.01 g g⁻¹) water contents (g H₂O g dry mass⁻¹), and slowest in seeds containing 0.04 g g⁻¹. A 45°C treatment before imbibition restored germination of dry seeds by melting crystallized triacylglycerols. Here, we show that the rate of deterioration in C. carthagenensis seeds stored at 5°C correlated with the rate that triacylglycerols crystallized within the seeds. Lipid crystallization, measured using differential scanning calorimetry, occurred at 6°C for this species and was fastest for seeds stored at 5°C that had high and very low water contents, and slowest for seeds containing 0.04 g g⁻¹. Germination decreased to 50% (P50) when between 16 and 38% of the triacylglycerols crystallized; complete crystallization took from 10 to over 200 days depending on water content. Our results demonstrate interactions between water and triacylglycerols in seeds: (1) water content affects the propensity of triacylglycerols to crystallize and (2) hydration of seed containing crystallized triacylglycerols is lethal. We suggest that these interactions form the basis of the syndrome of damage experienced when seeds with intermediate storage physiologies are placed in long-term storage.     

An oxidative burst of superoxide in embryonic axes of recalcitrant sweet chestnut seeds as induced by excision and desiccation

Recalcitrant seeds are intolerant of desiccation and cannot be stored in conventional seed banks. Cryopreservation allows storage of the germplasm of some recalcitrant seeded species, but application to a wide range of plant diversity is still limited. The present work aimed at understanding the stresses that accompany the first steps in cryopreservation protocols, wounding and desiccation, both of which are likely to lead to the formation of reactive oxygen species (ROS). Extracellular ROS production was studied in isolated embryonic axes of sweet chestnut ( Castanea sativa ). Axis excision was accompanied by a burst of superoxide (O₂^·−), demonstrated by a colorimetric assay using epinephrine, electron spin resonance and staining with nitroblue tetrazolium. Superoxide was immediately produced on the cut surface after isolation of the axis from the seed, with an initial 'burst' in the first 5 min. Isolated axes subjected to variable levels of desiccation stress showed a decrease in viability and vigour and increased electrolyte leakage, indicative of impaired membrane integrity. The pattern of O₂^·− production showed a typical Gaussian pattern in response to increasing desiccation stress. The results indicate a complex interaction between excision and subsequent drying and are discussed with a view of manipulating ROS production for optimisation of cryopreservation protocols.     

Developmental Changes in Relation to Desiccation Tolerance of Archontophoenix alexandrae (Palmae) Seeds

Germination of Archontophoenix alexandrae seeds and embryos were studied under gradient water content treatments throughout the seed development phases of maturation in 2005 to investigate seed desiccation tolerance and storage characteristics. During the maturation process, seed water content decreased gradually from55 DAF (days after flowering) to 70 DAF, and seeds reached the maximum dry-weight at 90 DAF. Seed germinability appeared after 60 DAF. Seeds germinated with a temperature range from15℃- 40℃ under alternating photoperiod (14 h light, 10 h dark, 12μmol m^{- 2}s ^{- 1} ), while the best germination percentage was obtained between 30℃- 35℃. A maximum germination capacity reached at 70 DAF. However, seed germination was greatly inhibited by light. Desiccation tolerance of seeds and embryos increasedgradually from 55 DAF to 90 DAF and reached the maximum at 90 DAF with a semilethal water content of 0.18 g/g ( seed) and 0.3 g/g ( embryo) respectively. Rapid dehydration maintained higher seed germination percentage than thatof slow dehydration when drying to the same water content. Seeds with without water content treatments failed to germinate after 1 month storage under - 18℃, whereas appropriate desiccation treatment prolonged seed longevity under 4℃, 10℃ and 15℃ storage temperatures. It revealed obviously the recalcitrant characteristics of Archontophoenix alexandrae seeds torage behaviour which are tolerant toward neither deep desiccation nor low temperatures.     

Influence of woodlot floor topography on microbial decomposer distribution

The distribution of microbial populations that decomposed sugar, cellulose and lignin-related substrates was examined in a beech Fagus grandifolia Ehrh. and maple Acer saccharum Marsh. dominated woodlot developed on glacial till. The topography of the woodlot, characterized by rises, depressions and more extensive level areas about 1 m in diameter with a 0.5 m vertical maximum, produced a mosaic of decomposer habitats designated as high, level and low sites.
In general, populations of sugar, cellulose and lignin decomposing organisms (based on ten estimates made from April to October) were two to four times higher in litter and soil samples from low sites than those from high sites. Sugar decomposing bacteria in litter were most abundant at all topographic sites. 135 × 10⁶ g⁻¹ dry litter at high sites, 396 × 10⁶ g⁻¹ at level sites and 456 × 10⁶ g⁻¹ at low sites; lignolytic fungi were least abundant, 391 × 10² g⁻¹ dry litter at high sites. 700 × 10⁶ g⁻¹ at level sites and 954 × 10² g⁻¹ at low sites. Numbers of microbial decomposers in the topographic sites were correlated with organic matter content. Distribution of fungal genera did not appear to be related to topographic site. Most populations examined showed two numerical peaks, one in late May or June and one in late September or October. It is suspected that these peaks were influenced by the coincident timing of favourable physical conditions and priming by soluble nutrients leached from litter.     

The effects of water hardness upon the uptake, accumulation and excretion of zinc in the brown trout, Salmo trutta L.

The effects of water hardness (9 and 220 mgl⁻¹ as CaCO₃) upon zinc exchange in brown trout exposed to 0.77 μmol Zn 1⁻¹ have been investigated using artificial soft water (<49.9 μmol Ca ^l-1, <40.1 μmol Mg 1⁻¹) and mains hard water (1671.7 μmol Ca 1⁻¹, 493.6 μmol Mg 1⁻¹) of known composition. Both hard and soft water-adapted fish exhibited a bimodal pattern of net zinc influx. Net zinc influxes during both fast and slow uptake phases were significantly greater ( P <0.001) in soft (82.9 and 6.2 μmol Zn 100 g⁻¹ h⁻¹) than in hard water (46.3 and 2.4 μmol Zn 100 g h⁻¹). Zinc efflux (- 0.2 μmol Zn 100 g⁻¹ h⁻¹) was enhanced only in hard water during the slow net influx phase.
Brown trout exposed to zinc in hard water and placed in metal-free media exhibited a greater net efflux (- 25.6 μmol Zn 100 g⁻¹ h⁻¹) of the metal than did fish in soft water (-4.2 μmol Zn 100 g⁻¹ h⁻¹) treated in the same manner. Tissue ⁶⁵Zn activities reflected both the differences in uptake and excretion rates of the metal between hard and soft water fish. During zinc exposure (0.77 μmol Zn 1⁻¹) high water hardness reduced tissue burdens of the metal by reducing net branchial influx, and enhancing efflux of the metal in hard water fish.