The effects of elevated temperature and humidity on rectal temperature and respiration rate in the New Zealand white rabbit

In 2 replicated factorial experiments, 7-h climate chamber exposures were used to study the responses of adult NZW rabbits to a range of elevated temperatures and humidities. At 18 mm Hg water vapour pressure, 23.8° C was well tolerated, rectal temperature (RT) and respiration rate (RR) averaging 38.6±0.3° C and 82.9±15.5 breaths/min, respectively. Both parameters were elevated (P<0.001) at 32.2°, 37.8° and 43.3° C. RT and RR reached plateau levels of 39.5–40.1° C and 410–460/min at 32.2° C, which was tolerated for the full 7-h test period. Test temperatures of 37.8° and 43.3° C, on the other hand, could be tolerated for only 80 and 40 min respectively, before RT reached the safe upper limit of 41.7° C. Final RR values at 37.8° and 43.3° C were 701.6±42.7 and 812±55.1/min, respectively. In a 34.5° C atmosphere a humidity of 21 mm Hg water vapour pressure was classified as dry, and was tolerated for 323±123 min. RT and RR increased by 0.6° C and 316/min during the first 20 min of exposure (P<0.05). Thereafter both parameters increased progressively, but with no significant differences between successive recording periods, until RR reached 550.3±88.8/min at 41.7° C RT. Humidities of 25, 29 and 33 mm Hg water vapour pressure were, on the other hand, classified as wet and were tolerated for only 92±22, 81±16 and 119±50 min, respectively. RR at the times that RT reached 41.7° C at these 3 humidities was 732±26, 789±30 and 764±23/min, respectively. The results point to the likelihood that thermal stress will adversely affect the productivity and welfare of NZW rabbits in the tropics unless adequate housing environments are provided. Significant between-individual phenotypic differences in heat tolerance suggest the need for genetic studies of the possibility of selecting for improved heat tolerance.     

Comparative glucose tolerance studies in the freshwater snail Biomphalaria glabrata: influence of starvation and infection with the trematode Schistosoma mansoni

Summary The dynamic reactions of B. glabrata on intravasal loads of 20–1000 g glucose·g live fresh wt^-1 were studied in standard fed snails (SFS). Starved snails (SS) and snails infected with S. mansoni (IS) were given 100–500 g glucose·g fresh live wt^-1. Mean hemolymph glucose level was 12.1 mg·100 ml^-1 in SFS. It was not significantly lower in SS (10.7 mg·100 ml^-1), but significantly reduced in IS (8.5 mg·100 ml^-1). Since hemolymph volumes were significantly increased in SS, the amount of circulating glucose (pool) did not change (75 g·g body weight^-1) compared to SFS (62 g·g^-1). It was, however, reduced to 41 g·g^-1 in IS. In SFS the circulating glucose pool had to be doubled to induce significantly elimination of the injected glucose. Tripled pools were eliminated with half-times of 45 min, whereas lower and higher glucose loads were eliminated significantly slower (half-times: 80–105 min). Glucose tolerance of SS was reduced: half-times were doubled, and metabolization of injected glucose was reduced. Since tissue fresh weights were lowered by 40%, absolute incorporation of ¹⁴C from labeled glucose was lowered, but specific incorporation (per mg) was higher than in SFS and IS. Glucose tolerance of IS was increased: metabolic clearance rates rose by 70% and half-times were shortened by 30%, though absolute and specific rates of ¹⁴C incorporation were lowered. However, IS lost 25% of the label to the water, whereas SFS lost 12% and SS only lost 8%. Using the antimetabolite 2-deoxyglucose, 80% of the losses proved to be glucose in IS, and 50% in SFS. The present results suggest the existence of a glucostatic regulation in B. glabrata with lower sensitivity and capacity than in mammals. As to glucose tolerance, the often reported parallelism in metabolic shifts induced by starvation and parasitic infection was not confirmed.Abbreviations SFS standard fed snails - SS starved snails - IS infected snails - MGG midgut-gland-gonad - RB rest of the bodymass - MCR metabolic clearance rate     

Development of an L6 myoblast in vitro model of moniliformin toxicosis

L6 myoblasts were used as an in vitro model to investigate the role of moniliformin and its interaction with monensin in turkey knockdown syndrome and sudden death syndromes in poultry. Cell viability and microscopic and ultrastructural alterations noted in L6 myoblasts cultured in the presence of moniliformin (0.0–0.3 g/l) were compared to those observed in parallel cultures also containing one of the following compounds: selenium (0–0.004 ng/l), thiamine (0–0.3 g/l), or pyruvate (0–0.46 g/l). Marked dilation of the RER, membranous whorls, glycogen deposition, membrane-bound cytoplasmic inclusions and necrosis were observed in myoblasts exposed to 0.03/2-0.30 g moniliformin/l medium. Supplementation of medium with thiamine and pyruvate, or selenium, provided significant protection to cells exposed to 0.0–0.3 g/l or 0.0–0.15 g moniliformin/l, respectively. Dose-dependent differences in protein and ATP production were not detected. Myoblasts grown in medium containing 0–0.15 g moniliformin/l and 7.5–50.0 M A23187, beauvericin or monensin had degrees of cytotoxicity similar to parallel cultures receiving only an ionophore. L6 myoblasts were a useful model of moniliformin toxicosis. The findings of this study suggest cytotoxicity due to moniliformin in L6 myoblasts may be due in part to oxidative damage and altered pyruvate metabolism, and that moniliformin does not predispose myoblasts to ionophore toxicosis. This study supports the results of in vivo investigations in poultry that moniliformin and monensin do not act synergistically to induce knockdown or monensin toxicosis.     

Photosynthesis and growth rates of Laurencia brongniartii J. Agardh (Rhodophyta, Ceramiales) in preparation for cultivation

Laurencia brongniartii is usually found at depths below 4 m, but can be found in shallow subtidal areas in crevices and on the walls of a coral reef in Amami Oshima Island, Kagoshima Prefecture, Japan, where irradiances were significantly lower than those at similar depths in open water. In preparation for the possible cultivation of this species for its antibiotic compounds, the effects of temperature and irradiance on photosynthesis and growth were measured. Photosynthesis and growth rates of L. brongniartii explants were highest at 26 and 28 °C, which closely corresponded to temperatures found during August to late December when it was most abundant. The estimated maximum photosynthesis rate (P _max) was 4.41 mol photon m^–2 s^–1 at 26 °C and 4.07 mol photon m^–2 s^–1 at 28 °C. Saturating irradiance occurred at 95 mol photon m^–2 s^–1 at 26 °C and 65 mol photon m^–2 s^–1 at 28 °C. In contrast, growth experiments at 41.7 mol photon m^–2 s^–1 caused bleaching of explants and the maximum growth rate observed during the study was 3.02 ± 0.75% day^–1 at 28 °C and 25 mol photon m^–2 s^–1. The difference in the saturating irradiance for photosynthesis and the irradiance that caused bleaching in growth experiments suggests that long-term exposure to high irradiance was detrimental and should be addressed before the initiation of large scale cultivation.     

Interspecific differences in aluminium tolerance in relation to root cation-exchange capacity

Genotypic differences in aluminium (Al) tolerance hold considerable promise in overcoming an important limitation to plant growth in acid soils. Little is known, however, about the biochemical basis of such differences. Extracellular properties, particularly low root cation-exchange capacity (CEC), have been associated with Al tolerance, since roots of low CEC adsorb less Al than do those of high CEC. A solution culture study was conducted in which 12 plant species (monocots and dicots) were grown in solution culture of low ionic strength (ca 2 mM) for 8 d at four Al concentrations (0, 16, 28 and 55 M). The species differed significantly in Al tolerance as shown by differences in root length. Root length relative to that of the same species grown in the absence of Al varied from 6 to 117% at 16 M Al, and from 6 to 75% at 28 M Al. Species tolerance of Al was not closely associated with differences in root CEC. Although in some species Al sensitivity was associated with high adsorption of Al during a 10- or 40-min exposure to Al (expressed on a fresh mass or root length basis), this was not a good predictor of Al tolerance across all species studied.     

Ecology of bacterial communities in the schistosomiasis vector snailBiomphalaria glabrata

The internal colony-forming bacterial flora of the schistosome intermediate host snailBiomphalaria glabrata (Say) has been characterized in ca. 500 individual snails from Puerto Rico, Guadeloupe, and St. Lucia, and from laboratory aquaria. Freshly captured wild snails harbor 2–40×10⁶ CFU·g^–1, and healthy aquarium snails harbor 4–16×10⁷ CFU·g^–1, whereas moribund individuals have 4–10 times as many bacteria as healthy individuals from the same habitats.Pseudomonas spp. are the most common predominant bacteria in normal snails, whereasAcinetobacter, Aeromonas, andMoraxella spp. predominate in moribund snails. External bacterial populations in water appear to have little effect on the composition and size of the flora in any snail. In addition to normal (healthy) and moribund snails, a third group of snails has been distinguished on the basis of internal bacterial density and predominating genera. These high-density snails may have undergone stresses and may harbor opportunistic pathogens. The microfloras of wild and laboratory-reared snails can be altered and stimulated to increase in density by crowding the snails or treating them with antibiotics.     

Effect of cooling rate on the survival of frozen wood frogs,Rana sylvatica

Summary Wood frogs (Rana sylvatica) were frozen to-2.5°C under five distinct cooling regimes to investigate the effect of cooling rate on survival. Frogs survived freezing when cooled at -0.16°C · h^-1 or -0.18°C · h^-1, but mortality resulted at higher rates (-0.30°C · h^-1,-1.03°C · h^-1, and -1.17°C · h^-1). Surviving frogs in the latter groups required longer periods to recover, and transient injury to the neuromuscular system was evident. Some of the frogs that died had patches of discolored, apparently necrotic skin; vascular damage, as indicated by hematoma, also occurred. It is concluded that slow cooling may be critical to the freeze tolerance of wood frogs. Additional studies examined the effect of cooling rate on physiological responses promoting freeze tolerance. Mean glucose concentrations measured in plasma (15–16 mol · ml^-1) and liver (42–45 mol · g^-1) following a 2-h thaw did not differ between slowly- and rapidly-cooled frogs but in both groups were elevated relative to unfrozen controls. Thus freezing injury to rapidly-cooled frogs apparently was not mitigated by the presence of elevated glucose. Water contents of liver tissue, measured 2 h post-thawing, did not differ between slowly-cooled (mean = 77.6%) and rapidly-cooled (mean = 78.5%) frogs. However, the mean hematocrit of slowly-cooled frogs (48%) was significantly higher than that (37%) of frogs cooled rapidly, possibly owing to differences in the dynamics of tissue water during freezing.     

Changes in temperature tolerance ofBalanus balanoides during its life-cycle

Summary 1. The barnacleBalanus balanoides exhibits little seasonal variation in upper lethal temperatures in North Wales.2. There are marked seasonal changes in resistance to sub-zero temperatures, the lower lethal varying from –6.0° C in June to –17.6° C in January.3. Exceptional tolerance to cold is acquired between December and January and is lost between February and April. Although these dates coincide with oviposition and naupliar liberation respectively, it was found that cold tolerance did not necessarily depend upon, or accompany, the normal breeding cycle.4. Cold tolerance was not acquired by animals kept cold in the laboratory during winter, nor was it lost in animals kept in the laboratory during spring. There was no evidence that changes in nutrition or in the light régime led to loss of cold tolerance.5. The cyprids were considerably less resistant to both high and low temperatures than the overwintering adults and the late-stage embryos. There was a marked increase in resistance at metamorphosis.6. The appearance of cold tolerance in the adult coincides with a period of physiological hibernation, involving loss of certain tissues, diminished feeding activity, respiration and biosynthesis. The metabolic inactivity of the animal may be a factor promoting the greatly increased tolerance to cold that we have observed, while the composition of the body fluids may also be modified during the winter in such a way as to protect the tissues.

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Veränderungen der Temperaturtoleranz vonBalanus balanoides während seines Lebenszyklus

Kurzfassung In Nordwales weisen die oberen Letaltemperaturen des CirripediersB. balanoides nur geringe jahreszeitliche Variationen auf. Jedoch treten je nach der Jahreszeit merkbare Resistenzveränderungen bei Temperaturen unter Null auf, wobei die untere Letaltemperatur von –6,0° C im Juni bis zu –17,6° C im Januar schwankt. Eine außergewöhnlich starke Kältetoleranz wird in der Zeit von Dezember und Januar erworben und zwischen Februar und April wieder verloren. Obwohl diese Zeitspanne mit der Oviposition beziehungsweise dem Schlüpfen der Nauplien zusammenfallen, konnte festgestellt werden, daß die Kältetoleranz nicht notwendigerweise vom Brutzyklus abhing oder diesen begleitete. Unter Laboratoriumsbedingungen wurde von kalt gehaltenen Tieren eine Kälteresistenz nicht erworben, auch ging diese bei Tieren, die während des Frühlings im Labor verblieben, nicht verloren. Es ließ sich nicht beweisen, daß Veränderungen in der Ernährung oder Änderungen in der Tageslänge zu einem Verlust der Kälteresistenz führen. Die Cypriden waren wesentlich weniger widerstandsfähig, sowohl gegenüber hohen wie niedrigen Temperaturen, als überwinternde Adulte und die ältesten Embryostadien. Während der Metamorphose zeigte sich eine merkliche Erhöhung der Temperaturresistenz. Das Auftreten der Kälteresistenz beim Adultus fiel mit einer Periode physiologischen Winterschlafs zusammen, wobei gewisse Gewebe reduziert wurden und Nahrungsaufnahme, Atmung und biosynthetische Aktivität nachließen. Dieser stoffwechselphysiologische Aktivitätsrückgang könnte ein Faktor sein, der die beobachtete erhöhte Kältetoleranz fördert. Außerdem wird möglicherweise auch die Zusammensetzung der Körperflüssigkeiten während des Winters so verändert, daß die Gewebe geschützt werden.

     

Factors affecting in vitro microrhizome production in turmeric

In vitro microrhizome production was obtained in turmeric (Curcuma longa Linn.). Freshly sprouted buds with small rhizome portions excised from stored mature rhizomes were cultured on semi-solid culture initiation medium –- MS basal medium + 0.88 M BAP (6-benzylaminopurine) + 0.92 M kinetin + 5% coconut water + 2% sucrose + 0.5% agar –- resulting in bud elongation. Multiple shoots were produced from these elongated buds by culturing in liquid shoot multiplication medium –- MS basal medium + 2.2 M BAP + 0.92 M kinetin + 5% coconut water + 2% sucrose –- at 25±1°C and 16-h light (at 11.7 mol m^–2 s^–1)/8-h dark cycles. Clumps of four to five multiple shoots/single shoots were used in various experiments. Cultures were incubated in the dark at 25±1°C. Half strength MS basal medium supplemented with 80 g l^–1 sucrose was found to be optimal for microrhizome production. Cytokinin BAP had an inhibitory effect on microrhizome production. At the highest concentration of BAP tried (35.2 M) microrhizome production was totally inhibited. Microrhizome production depended on the size of the multiple shoots used. Microrhizomes produced were of a wide range in size (0.1–2.0 g) and, readily regenerated when isolated and cultured in vitro on culture initiation medium or shoot multiplication medium. Under in vivo conditions, small (0.1–0.4 g), medium (0.41–0.8 g) and big (>0.81 g) microrhizomes regenerated. Plantlets developed from big microrhizomes grew faster.     

Uptake of sewage derived nitrogen by<Emphasis Type="Italic"> Ulva rigida</Emphasis> (Chlorophyceae) in Bahía Nueva (Golfo Nuevo,Patagonia, Argentine)

Uptake kinetics of nitrogen derived from sewage–seawater mixtures (2.5–20% v/v effluent) were determined in the laboratory for Ulva rigida (Chlorophyceae) native from Bahía Nueva (Golfo Nuevo, Patagonia, Argentine). In terms of nitrogen concentration, experimental enrichment levels varied between 53.7 and 362.3M of ammonium and between 0.77 and 6.21M of nitrate+nitrite. Uptake rates were fitted to the Michaelis–Menten equation, with the following kinetic parameters: ammonium: V_max = 591.2molg^–1h^–1, K _s=262.3M, nitrate+nitrite: V _max=12.9molg^–1h^–1, K _s=3.5M). Both nutrients were taken up simultaneously, but ammonium incorporation was faster in all cases. The results show a high capability of Ulva rigida to remove sewage-derived nitrogen from culture media. In the field, most of the nitrogen provided by the effluent would be tied up in algal biomass, supporting low nitrogen levels found at a short distance away from the source.     

A comparison of proline, thiol levels and GAPDH activity in cyanobacteria of different origins facing temperature-stress

Three cyanobacterial strains originating from different habitats were subjected to temperature shift exposures and monitored for levels of proline, thiol and activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Thermophile Mastigocladus laminosus (growth optimum, 40 °C), raised the proline level 4.2-fold at low temperature (20 °C), for the psychrophile Nostoc 593 (growth optimum, 20 °C), it was raised 8-fold at 40 °C while in the mesophile Nostoc muscorum (growth optimum, 30 °C), the imino acid level increased 2.3-fold during temperature shiftdown to 20 °C or 3.5-fold in sets facing shiftup (40 °C). Alterations in thiol levels in the above strains were in line with proline. It is suggested that such fluctuations reflect metabolic shifts as a response to stress. Interestingly, GAPDH activity was maximum at the respective growth temperature optimum of M. laminosus (122 nmol NADPH oxidized min ^–1 mg ^–1 protein) and Nostoc 593 (141 nmol NADPH oxidized min ^–1 mg ^–1 protein) while in N. muscorum, it increased at 40 °C (101 nmol NADPH oxidized min ^–1 mg ^–1 protein) and to 93.3 nmol NADPH oxidized min ^–1 mg ^–1 protein (20 °C) relative to 86 nmol NADPH oxidized min ^–1 mg ^–1 protein at 30 °C. It seems that extremophiles maintain the GAPDH activity/level during growth at their respective temperatures optimal while the mesophile increases it in order to cope up with temperature-stress.     

The productivity of mesocyclops leuckrarti (Claus) in Lake Kinneret (Israel)

Specific rates of growth (Cw) of Mesocyclops leuckarti, young male and female instars varied between 0.03–0.26 and 0.03–0.17 g_(w.w.)/g_(w.w.)/day respectively at 15° and 22°C, whilst at IV–V copepodid stages females showed a higher Cw values. During 1969–1975 the averages of productivity and monthly P/B ratios were 44 (±23) g_(w.w.) (= 5g_(w.w.)/m²/month) and 3.1 respectively. P/B ratios were highly correlated (r(sup2)=0.98) with temperature changes. Metabolic parameters and P/B ratios were found to be similar to other water bodies in the world indicating an adaptation of M. leuckarti to different conditions.     

Selection of amitrole tolerant tobacco calli and the expression of this tolerance in regenerated plants and progeny

Summary Thirty-one clones capable of growth in the presence of 1.9×10^–4 M amitrole (3-amino-1,2,4-triazole) were isolated from non-mutagenized cell suspensions of haploid Nicotiana tabacum cv. Wisconsin 38 plants at a frequency of 2.5×10^–8. Seven clones retained tolerance when grown on selective medium for three years. When clones were cultured in the absence of amitrole, tolerance persisted for 9 months in five clones. Some plants regenerated from three amitrole-tolerant clones were tolerant. Seven amitrole-tolerant clones were isolated from diploid N. tabacum cell suspensions and R plant tolerance was followed through two sexual generations. Simple Mendelian inheritance patterns were not observed.     

Cadmium tolerance in tobacco cell culture and its relevance to temperature stress

Growth of unselected tobacco (Nicotiana tabacum W38) cell suspension cultures was reduced by 50–200 M cadmium (Cd) in the culture medium and cells were killed by 400 M Cd. Tolerance to Cd was increased either by using rapidly growing cells or by culturing cells at higher densities. Cell lines tolerant to 2 mM Cd were established by progressively elevating levels of Cd in the culture medium. The Cd tolerance was not due to differences in uptake between unselected and Cd-tolerant cell lines, and the tolerance to Cd was not lost during long term culture in the absence of Cd. Cd-tolerant cells also showed higher tolerance to heat shock (37.5°C, 2–8 hours) and cold treatments (4°C, 1–7 days) than the unselected cells.     

Differences in heat tolerance plasticity between supratidal and intertidal snails indicate complex responses to microhabitat temperature variation

Tropical intertidal gastropods that experience extreme and highly variable daily temperatures have evolved significant and complex heat tolerance plasticity, comprising components that respond to different timescales of temperature variation. An earlier study showed different plasticity attributes in snails from differently-heated coastlines, suggesting lifelong irreversible responses that matched habitat thermal regimes. To determine whether heat tolerance plasticity varied at a finer, within-shore spatial scale, we compared the responses of supratidal (predominantly shade-dwelling) and intertidal (frequently solar-exposed) populations of the tropical thermophilic gastropod, Echinolittorina malaccana. Snails modified lethal temperature (LT₅₀) under warm or cool laboratory acclimation, with the overall variation in LT₅₀ being greater in the supratidal (56.0–58.0 °C) than in the intertidal population (57.1–58.1 °C). Similar maximum LT₅₀s expressed by the populations after warm acclimation suggest a capacity limitation under these temperature conditons. The different minimum LT₅₀s after cool acclimation corresponded with microhabitat temperature and field acclimatization of the snails. Different responses to the same laboratory acclimation treatment imply long-term (and possibly lifelong) thermal acclimatization, which could benefit sedentary organisms that are randomly recruited as larvae from a common thermally-stable aquatic environment to thermally-unpredictable intertidal microhabitats. These findings provide another example of thermal tolerance plasticity operating at microhabitat scales, suggesting the importance of considering microhabitat thermal responses when assessing broad-scale environmental change.     

The responsiveness of Avena fatua aleurone protoplasts to gibberellic acid

The sensitivity to gibberellic acid (GA₃) of aleurone protoplasts isolated from a single harvest of an inbred line of Avena fatua seed that had been after-ripened over anhydrous CaCl₂ at 25±2°C and 4±2°C for three years was assessed. Protoplasts isolated from aleurones of seed stored at 25°C produced substantially more -amylase in response to 10^–7 M GA₃ than those isolated from aleurones of seed stored at 4°C. The apparent difference in responsiveness does not appear to be due to a change in the duration of the lag phase between addition of GA₃ and the production of -amylase. The dose response of aleurone protoplasts to GA₃, measured as -amylase production, is complex and appears to have three phases. Protoplasts from seed stored at both temperatures respond appreciably to 10^–14 M GA₃. With increasing concentrations of GA₃, up to 10^–9 M, -amylase production increases similarly in protoplasts from both lots of seed, reaching a level approximately 2.7–3.8 times greater than when no GA₃ is applied. GA₃-induced -amylase production increases markedly as the concentration is raised from 10^–9 M to 10^–6 M, and the response then appears to be saturated. Over this part of the response curve protoplasts from the two seed lots differ markedly in their responsiveness to GA₃. Those from seed stored at 25°C produce considerably more -amylase, >130-fold higher than the minus GA₃ control, than those from seed stored at 4°C, <35-fold higher than the minus GA₃ control. This apparent difference in the responsiveness of aleurone protoplasts to GA₃ could be correlated with the loss of embryo dormancy in seed stored at 25°C. Seed stored at 4°C retained the dormancy characteristics present immediately after harvesting.     

Interrelationships Between Nitrogen Supply and Photosynthetic Parameters in Vicia faba L.

We determined for Vicia faba L the influence of nitrogen uptake and accumulation on the values of photon saturated net photosynthetic rate (P _Nmax), quantum yield efficiency (), intercellular CO₂ concentration (C _i), and carboxylation efficiency (C _e). As leaf nitrogen content (N_L) increased, the converged onto a maximum asymptotic value of 0.0664±0.0049 mol(CO₂) mol(quantum)^–1. Also, as N_L increased the C _i value fell to an asymptotic minimum of 115.80±1.59 mol mol^–1, and C _e converged onto a maximum asymptotic value of 1.645±0.054 mol(CO₂) m^–2 s^–1 Pa^–1 and declined to zero at a N_L-intercept equal to 0.596±0.096 g(N) m^–2. fell to zero for an N_L-intercept of 0.660±0.052 g(N) m^–2. As N_L increased, the value of P _Nmax converged onto a maximum asymptotic value of 33.400±2.563 mol(CO₂) m^–2 s^–1. P _N fell to zero for an N_L-intercept of 0.710±0.035 g(N) m^–2. Under variable daily meteorological conditions the values for N_L, specific leaf area (_L), root mass fraction (R_f), P _Nmax, and remained constant for a given N supply. A monotonic decline in the steady-state value of R_f occurred with increasing N supply. _L increased with increasing N supply or with increasing N_L.     

An abscisic acid analog inhibits abscisic acid-induced freezing tolerance and protein accumulation, but not abscisic acid-induced sucrose uptake in a bromegrass (Bromus inermis Leyss) cell culture

The application of abscisic acid (ABA), either as a racemic mixture or as optically resolved isomers, increases freezing tolerance in a bromegrass (Bromus inermis Leyss) cell culture and induces the accumulation of several heat-stable proteins. Two stereoisomers of an ABA analog, 23 dihydroacetylenic abscisyl alcohol (DHA), were used to study the role of ABA-induced processes in the acquisition of freezing tolerance in these cells. Freezing tolerance was unchanged in the presence of (–) DHA (LT₅₀ -9°C), and no increase in heat-stable protein accumulation was detected; however, the (+) enantiomer increased the freezing tolerance (LT₅₀ -13°C) and induced the accumulation of these polypeptides. All three forms of ABA increased freezing tolerance in the bromegrass cells, although (–) ABA was less effective than either (+) or (±) ABA when added at equal concentrations. Cells pretreated with 20 or 50 M (–) DHA displayed lower levels of freezing tolerance following the addition of 2.5, 7.5 or 25 M (±) ABA. Full freezing tolerance could be restored by increasing the concentration of (±) ABA to > 25 M. Pretreatment of cells with (–) DHA (20 or 50 M) had no effect on freezing tolerance when 25 M (+) ABA was added. The induction of freezing tolerance by 25 M (–) ABA was completely inhibited by the presence of 20 M (–) DHA. The accumulation of ABA-responsive heat-stable proteins was inhibited by pretreatment with 20 M (–) DHA in cells treated with 2.5 or 7.5M (+) ABA, and in cells treated with 25 M (–) ABA. The accumulation of these polypeptides was restored when (±) or (+) ABA was added at a concentration of 25 M. The analysis of proteins which cross-reacted with a dehydrin antibody revealed a similar inhibitory pattern as seen with the other ABA-responsive proteins. The effects of the various isomers of ABA and DHA on cell osmolarity and sucrose uptake was also investigated. In both cases, (±) and (+) ABA had pronounced effects on the parameters measured, whereas (–) ABA treated cells gave substantially different results. In both sucrose uptake and cell osmolarity, DHA had no significant effect on the results obtained following (±) or (+) ABA treatment. Maximum freezing tolerance was only observed in cells when both heat-stable protein accumulation and sucrose uptake were observed.Abbreviations ABA abscisic acid - DHA 2,3 dihydroacetylenicabscisyl alcohols - DMSO dimethyl sulfoxide - LT₅₀ temperature at which 50% of cells are killed The authors would like to acknowledge the technical assistance of Angela Bollman, Bruce Ewan and Angela Shaw. This work was supported by grants from the Natural Science and Engineering Research Council of Canada to L.V.G. and N.H.L., and a grant from the University of Saskatchewan to R.W.W.     

Photosynthetic performance of transplanted ecotypes ofEcklonia cava(Laminariales, Phaeophyta)

Young sporophytes of short-stipe ecotype ofEcklonia cavafrom a warmer locality (Tei, Kochi Pref., southern Japan) and those of long-stipe ecotype from a cooler locality (Nabeta, Shizuoka Pref., central Japan) were transplanted in 1995 to artificial reefs immersed at the habitat of long-stipe ecotype in Nabeta Bay, Shizuoka Pref., central Japan. The characteristics of photosynthesis and respiration of bladelets of the transplanted sporophytes of the two ecotypes were compared in winter and summer 1997; the results were assessed per unit area, per unit chlorophyllacontent and per unit dry weight. In photosynthesis-light curves at 10–29 °C, light saturation occurred at 200–400 mol photon m^–2s^–1in sporophytes from both Tei and Nabeta. The maximum photosynthetic rate (P _max) at 10–29 °C and the light-saturation index (I _k) at 25–29 °C in sporophytes from both localities were generally higher in winter than in summer.P _maxat 25–29 °C (per unit area and chlorophylla) were higher in sporophytes from Tei than those from Nabeta in both seasons. The optimum temperature for photosynthesis was 25 °C in winter and 27 °C in summer at high light intensities of 100–400 mol photon m^–2s^–1. However, at lower light intensities of 12.5–50 mol photon m^–2s^–1, it was 20 °C in winter and 25–27 °C in summer for sporophytes from both locations. Dark respiration increased with temperature rise in the range of 10–29 °C in sporophytes from both locations in summer and winter. The sporophytes transplanted from Tei (warmer area) showed higher photosynthetic activities than those from Nabeta (cooler area) at warmer temperatures even under the same environmental conditions. This indicates that these physiological ecotypes have arisen from genetic differentiation.     

Mercury accumulation in sediment cores and along food chains in two regions of the Brazilian Pantanal

The Pantanal is a 140,000 km² floodplain wetland stretching acrosswestern Brazil and parts of Bolivia and Paraguay. Gold mining withmercury (Hg) amalgamation has thrived since 1980 along its northern rim. We quantified Hg accumulation in sediment cores (N = 5) and food chainsin this general region of the northern Pantanal and in a reference region,200 km deeper into the wetland (Acurizal). Cores were dated with²¹⁰Pb and ¹³⁷Cs using direct gamma-assay. Total Hg wasanalyzed by cold-vapor atomic fluorescence using a gold-meshpre-concentration trap. Average pre-1940 Hg accumulation in cores wasnot significantly different (N = 5, p= 0.14) between both regions andcomparable with rates calculated for global reference sites. Post gold-rushHg (post-1980) deposition averaged 55 ± 11.3 g m^-2yr^-1 in the northern impacted region and was more than 1.5 timeshigher than the post-1980 rate in Acurizal, implying a regional Hg effectof gold mining. Post-1980 Hg accumulation in Acurizal, in turn, was 2.1times the rate reported for a global reference during that time period,suggesting an additional basin-wide effect over such reference sites. Bycombining our core data with assessments of the size of the impacted areaand the amount of Hg released to the region since 1980, we estimated thatonly 2–8% of this Hg was recovered as a sedimentary signal. Theremainder of the Hg was lost to the atmosphere, downstream areas, orstored in biota. Hg concentrations in surface sediments in the northernPantanal (45.5 ± 5.5 ng g_dry ^-1) were significantlyhigher than those in our reference region (29.1 ± 0.7ng g_dry ^-1). Hg levels in primary producers were alsoelevated in the northern Pantanal. Eichhornia crassipes rootscontained 2.7–3.0 times more mercury than shoots in both regions and Salvinia auriculata, suggested as a biological monitor for Hg pollution,contained almost four times more mercury in the northern Pantanal (90.7± 9.1 ng g_dry ^-1) than in Acurizal (24.5 ± 3.3ng g_dry ^-1). Plant grazers and scavengers, such as apple snails(Pomacea sp.) and adult water beetles (Fam. Hydrophilidae), werelow in Hg, confirming previous data showing that the channeling of mercuryfrom lower to higher trophic levels along herbivorous links was inefficientcompared to transfer along carnivorous links. Collections of 12–16individuals of four species of Characidae (Aphyocharax sp., Tetragonopterus sp., Serrasalmus spiropleura and Pygocentrisnattereri) in both regions showed elevated Hg body burdens in bothpiranhas S. spiropleura and P. nattereri from the northernPantanal (149.9 ± 84.2 and 302.2 ± 159.1ng g_dry ^-1, respectively). Fish length for each species was notstatistically different between regions. P. nattereri length correlatedsignificantly (p<0.001) with Hg content in both regions, but the slopeof the regression in the northern Pantanal was 2.6 times the slope for theAcurizal collection, indicating an elevated rate of biomagnification in theHg-impacted region. Signals of Hg use in mining can be quantified insediment core chronologies and biological tissues, although species atdifferent trophic levels show dissimilar impacts. Mechanisms involved in Hgmagnification along food chains deserve more attention, particularly intropical regions where the threat of chronic exposure to this neurotoxinmay have the greatest implications for biodiversity.