Freeze-induced phase transitions in the plasma membrane of isolated protoplasts

Protoplasts were enzymically isolated from 2-week-old non-acclimated rye ( Secale cereale L. cv. Puma) seedlings. They were resuspended in isotonic sorbitol with different concentrations (0–10%) of dimethyl sulfoxide (DMSO). The survival of the protoplasts frozen in isotonic sorbitol solutions declined at temperatures below the freezing point with the LT₅₀ being -8°C. Addition of DMSO to the osmoticum increased survival at freezing temperatures. The optimum concentration of DMSO was 4% and lowered the LT₅₀ to -19°C. Freeze-fracture studies of the plasma membrane revealed aparticulate lipid lamellae at -4°C, but the first appearance of lateral phase separations, striations and inverted cylindrical micelles (hexagonal₁₁-type structures) occurred at -6°C. At lower temperatures, -8 and -10°C, the occurrence of nonbilayer structures became more common. The addition of DMSO decreased the incidence of the ultrastructural changes. With 2 or 4% DMSO, non-bilayer structures were not observed at temperatures above -10°C. Instead, striations and H₁₁-type structures were observed at - 15 and -20°C.     

Insecticidal activity influence of destruxins on the pathogenicity of Paecilomyces javanicus against Spodoptera litura

Abstract:  The bioactivities of destruxins (dtx), depsipeptides isolated from Metarhizium anisopliae , against Spodoptera litura were tested in laboratory. For contacting toxicities, dtx-E was more effective than dtx-A and dtx-B. The LC₅₀s values of dtx-A, B and E were 197.98, 292.00 and 113.99 mg/l at 48 h after treatment, while the LT₅₀s were 42.65, 59.45 and 23.68 h at 300 mg/l. In the experiment of antifeedant activity, dtx-A, dtx-B and dtx-E at five concentrations (200, 100, 50, 25 and 12.5 mg/l) were bioassayed. Destruxins in a dose-dependent manner gave an apparent antifeedant activity. Generally, dtx-A, over dtx-B and dtx-E had the significant (P < 0.05) larger choice and no-choice antifeedant indexes (CAIs and NCAIs). At the concentration of 200 mg/l, the CAIs or NCAIs of dtx-A, dtx-B and dtx-E were 96.78, 84.93 and 85.90 or 89.75, 62.42 and 72.28 respectively. Furthermore, the synergistic activity of crude destruxin (CD) for pathogenicity of Paecilomyces javanicus strain Pj01 was detected. The LC₅₀s values of single Pj01 and the mixtures of Pj01 plus CD at 100 or 200 mg/l (Pj01-CD100 or Pj01-CD200) were respectively 474.63 × 10⁵, and 197.45 × 10⁵ or 113.11 × 10⁵ spores/ml at the fifth day after treated. Meanwhile, Pj01, Pj01-CD100 and Pj01-CD200 gave the LT₅₀s values of 6.99 day, 5.49 day and 4.21 day at 100 × 10⁵ spores/ml. Clearly, dtx decreased the values of LC₅₀ and LT₅₀ of the strain Pj01.     

Intergenerational acclimation in aphid overwintering

Abstract.  1. When first instar nymphs and adults of the grain aphid Sitobion avenae (Fabricius) (Hemiptera: Aphidiae) were maintained in long-term cultures (>6 months) at 20 °C and 10 °C, the LT₅₀ decreased from −8 and −8.8 °C to −16.0 and −13.5 °C, respectively.
2. When aphids from the 20 °C culture were transferred to 10 °C, there was a progressive increase in cold tolerance through three successive generations. Transfer of newly moulted pre-reproductive adults reared at 10 °C for three generations back to 20 °C resulted in a rapid loss of cold hardiness in their nymphal offspring.
3. In all generations reared at 10 °C, first born nymphs were more cold hardy than those born later in the birth sequence. The LT₅₀ of nymphs produced on the first day of reproduction in the first, second and third generations maintained at 10 °C were −14.8, −17.0 and −16.6 °C, respectively. Thereafter, nymphal cold hardiness decreased over the subsequent 14 days of reproduction in each generation at 10 °C with mean LT₅₀ values of −10.3, −12.6 and −14.8 °C, respectively. By contrast, the cold tolerance of first born nymphs of aphids reared continuously at 20 °C did not differ in comparison with later born siblings. The LT₅₀ of adult aphids was also unaffected by ageing.
4. The ecological relevance of these findings is discussed in relation to the overwintering survival of aphids such as S. avenae .     

Drought stress-induced changes in the composition and physical state of phospholipids in wheat

Seedlings of winter wheat ( Triticum aestivum L. cv. Jubilejnaja 50) were grown under normal and dry conditions. Frost resistance (LT₅₀) of 10-day-old control seedlings was −6°C. LT₅₀ of the subsequently drought-stressed leaves shifted to −16°C. In plants of the same physiological age (28 days) but grown without stress, LT₅₀ was −12°C. Phosphatidylcholine accumulated and phosphatidylethanolamine decreased in drought-stressed leaves. Fatty acid unsaturation of these phospholipids increased with leaf age, independently of water supply. Both ageing and drought stress produced a decrease in the apparent phase separation temperature of isolated total phospholipids as determined by electron spin resonance. The possible role of dehydration-induced structural changes in the bilayer matrix in triggering adaptive alterations in membrane composition, similar to those observed during cold hardening, is discussed.     

Differential cold hardiness in adults and nymphs of the peach-potato aphid Myzus persicae

The LT₅₀ (lethal temperature) of first instar and adult stages of the peach-potato aphid Myzus persicae was lowered following long term acclimation at low temperatures.
First instars consistently showed greater cold hardiness than adult stages at each acclimation temperature, with the differential increasing as the temperature was lowered. When maintained at 5°C (the lowest acclimation regime) nymphs and adults had dLT₅₀8.3°C and 4.7°C respectively lower than those for non-acclimated individuals.
When 10°C acclimated adults were returned to 20°C, the acclimation effect was retained in full for 6 days but complete deacclimation occurred by day 10. In contrast the LT₅₀ of their progeny increased gradually from the first day of adult deacclimation towards the level of the unacclimated control over a period of 10 days.
A change in cold hardiness was observed in first instars according to their position in the birth sequence. The LT₅₀ of first-born nymphs (day 1 of reproduction) from 20°C parents was - 15.9°C rising to - 8.3°C by day 4 and remaining at this level until the end of the reproductive period.
The differential mortality between nymphs and adults observed in the laboratory was supported by the results of a field experiment. Adult aphids kept in clip-cages on a crop of oilseed rape showed greater mortality compared with those introduced as nymphs when the minimum temperature fell below -4°C for the first time in winter. At - 10°C mortality of aphids introduced as adults approached 100% whereas more than 50% of those introduced as nymphs were still alive at this temperature.     

Seasonal variation in freezing tolerance of the New Zealand alpine cockroach Celatoblatta quinquemaculata

1. The cold hardiness of the alpine cockroach Celatoblatta quinquemaculata was investigated. This species is found at 1360 m a.s.l. beneath schist slabs on the Rock and Pillar Range (Central Otago, New Zealand). Cockroaches were collected monthly from January to December 1996, and their LT₅₀ and supercooling points determined.
2. Celatoblatta quinquemaculata was freezing tolerant throughout the year, with a lower lethal temperature in winter of – 8.9 °C. Celatoblatta quinquemaculata was also found frozen under rocks in the field when the under-rock temperature was below – 3 °C, and could survive being frozen at – 5 °C for 4 days in the laboratory.
3. There was a marked decrease in LT₅₀ temperature from – 5.5 °C in April to – 7.5 °C in May. This coincides with decreasing temperatures from summer through autumn to winter, during which temperatures beneath snow-covered rocks may reach – 7.3 °C.
4. Supercooling points fluctuated during the year, with an increase from – 4.2 °C in autumn to – 3.4 °C in winter. Supercooling point was highest in spring, and changes in supercooling point do not appear to be related to changes in LT₅₀.
5. Recordings of environmental temperatures from the Rock and Pillar Range suggest that cockroaches may undergo up to twenty-three freeze–thaw cycles in the coldest month of the year, and that they may remain frozen for periods of up to 21 h. Maximum cooling rates recorded in the field (0.01 °C min^–1) were 100-fold slower than laboratory cooling rates, so survival estimates from laboratory experiments may be underestimates.     

Cold-acclimation induced changes in freezing tolerance and translatable RNA content in Citrus grandis and Poncirus trifoliata

Cold-acclimation-induced changes in freezing tolerance and translatable RNA content were compared in seedlings of a relatively cold sensitive citrus species, Citrus grandis L. Osb. cv. Thong Dee (pummelo), and the cold-hardy citrus relative, Poncirus trifoliata L. Raf. cv. Pomeroy (trifoliate orange). Cold acclimation of pummelo (10 days at 15°C followed by 4 weeks at 10°/5°C, day/night) resulted in a decrease in LT₅₀ from −6 to −8°C, while in trifoliate orange (acclimated for 7 weeks at 5°C), the LT₅₀ decreased from −9 to −18°C. Qualitative changes in the in vitro translation profile, revealed by two-dimensional gel electrophoresis, were observed following cold acclimation in both species. An mRNA for a large polypeptide (ca 160 kDa) was detected following cold acclimation of trifoliate orange. A similar change was not observed in pummelo following cold acclimation.     

Cold-induced changes in the polysome pattern and protein synthesis in winter rye (Secale cereale) leaves

Cold-induced changes in the polysome pattern and protein synthesis were analyzed in winter rye, Secale cereale L. cv. Voima, during one week's cold stress treatment, which was performed by transferring the 7-day-old plants from the greenhouse (25°C, long-day conditions) to 3°C and a photoperiod of 10. 5 h. Freezing resistance determined by electrolyte leakage increased significantly upon cold stress starting from LT₅₀ value –5°C. and reaching –9°C on the day 7 of cold exposure. After 4 weeks at low temperature, plants reached an LT₅₀ of –12°C. The polysome content increased markedly during cold stress compared to the control plants. After 2 weeks of cold treatment the polysome content decreased to the same level as that in control plants. The size-class distribution of polysomes showed a high proportion of large protein synthesizing polysomes in cold-stressed plants. After 2 weeks the values were comparable to those in control plants. Cold-induced proteins were detected using ³⁵S-labelled methionine for in vitro translations. At least 2 new polypeptides, M_r 30000 and 18000, were induced on the first day of cold stress and continued to be expressed at low temperatures 4 weeks later.     

THE COMBINED EFFECTS OF LOW TEMPERATURE AND SO2+NO2 POLLUTION ON THE NEW SEASON'S GROWTH AND WATER RELATIONS OF PICEA SITCHENSIS

Picea sitchensis (Bong.) Carr. seedlings were exposed to SO₂, NO₂ and SO₂+ NO₂ during dormancy in controlled environments, and were taken to night temperatures of 4, 0, −5, −10 and −15 °C in a freezer. Conditions in the freezer were carefully monitored during the low–temperature treatments. In two experiments, different photoenvironments and temperature regimes were imposed prior to the cold treatments, and different effects were observed. In the first, only limited frost hardiness was achieved and night temperatures of −15 °C were lethal. Temperatures of −5 and − 10 °C led to poor survival of lateral buds, particularly in plants exposed to 45 ppb SO₂. The poor bud break in plants exposed to SO₂ and to − 5 °C resulted in a loss of the effectiveness of this temperature as a chill requirement. Pressure-volume analysis showed that the shoots of plants exposed to NO₂ had greater elasticity (lower elastic moduli, e), so that loss of turgor occurred at lower relative water contents. In contrast, a hardening period (2 weeks in night/day temperatures of 3/10 °C and 8 h days at 50 μmol m⁻² s⁻¹ PAR) gave decreased elasticity and lower solute potentials of spruce shoots. In the second experiment, exposure to 30 ppb SO₂ and SO₂+ NO₂ led to slight, but consistent, increases in frost injury to the needles of plants frozen to − 5 and − 10 °C. The results suggest that the main interaction of low temperatures and winter pollutants may be on bud survival rather than on needle damage, but that effects are subtle, only occurring with certain combinations of pollutant dose and cold treatment.     

Role of cell wall in freezing tolerance of cultured potato cells and their protoplasts

Cultured potato ( Solanum tuberosum L., cv. Red Pontiac) cells suspended in PEG 1000 solutions of 0.6 and O.S osmol exhibited significantly different freezing tolerance from the same cells when suspended in PEG 6000 solutions of the same osmolalities. Cells suspended in PEG 6000 showed cytorhysis instead of plasmolysis. Cells in 0.2 and 0.4 osmol PEG 1000 had LT₅₀(1 of −2.5°C, but the LT₅₀ decreased to −7.50C as the osmolality increased to 0.8 osmol. In PEG 6000 the LT₅₀ remained at −2.50C for all osmolalities used, up to and including 0.8 osmol.
Released protoplasts suspended in 0.5 M sucrose had LT₅₀ of −21.5°C, compared to −12°C for whole cells suspended in the same medium. These results lend credence to an involvement of the cell wall in freezing injury of cultured potato cells, and are interpreted in terms of the generation of a mechanical stress between cell wall and plasma membrane during the freeze-thaw cycle.     

Contact with the host plant enhances aphid survival at low temperatures

1. The hypothesis, suggested by previous studies, that host plant contact reduces the cold tolerance of anholocyclic aphids was tested under laboratory conditions. Adult and first-instar Rhopalosiphum padi (L.) were exposed to temperatures of 0, –5 and –10 °C on intact plants, excised leaves and in the absence of contact with plant material.
2. Median lethal time (LT₅₀) values at all three temperatures indicate that aphids exposed in association with plant material survive longer than aphids that have no contact with their host plant. The difference in survival was most pronounced at –10 °C. Therefore, the above hypothesis is rejected for aphids on cereals because host plant contact apparently enhances cold tolerance.
3. Exposure on excised leaves also enhanced aphid survival at low temperature but was less effective than the intact plant. This suggests that plant quality as well as the presence or absence of plants is important in the cold tolerance of aphids on cereals.     

Inositol 1,4,5-trisphosphate formation in leaves of winter oilseed rape plants in response to freezing, tissue water potential aed abscisic acid

Time courses of formation of inositol 1,4,5-trisphosphate (IP₃) were followed in the leaves of non-acclimated and cold (2°C)-acclimated winter oilseed rape ( Brassica napus L. var. oleifera ) plants, subjected to different freezing temperatures or to polyethylene glycol 8000 (PEG) and abscisic acid (ABA) treatments. Changes in water potential (Ψ_w) and in ABA level in the frost- and PEG-treated tissues were also determined. Results obtained indicate that temperatures sligthly higher than LT₅₀ induced a transient and substantial increase in IP₃ level, both in non-acclimated and cold-acclimated tissues. At comparable freezing temperature (–5°C) the response of cold-acclimated leaves was lower than that of non-acclimated ones. The PEG-depedent decrease in Ψ_w to –0.9 MPa or ABA (0.1 m M ) treatment gave rise to a transient increase in IP₃ content in non-acclimated tissues only. Collectively, the data indicate that cold acclimation of plants may lead to lower cell responsiveness to the factors studied in terms of induction of IP₃ formation. Changes in the IP₃ content, observed in the present experiments, support our previous suggestion that non-killing freezing temperatures may induce the phosphoinositide pathway, both in non-acclimated and cold-acclimated tissues. Lowering of tissue water potential to some threshold value or a high exogenous ABA supply may mimic the freezing-dependent reaction in the non-acclimated leaves.     

Reduced susceptibility of Anopheles gambiae to permethrin associated with the use of permethrin-impregnated bednets and curtains in Kenya

Abstract. Susceptibility of the malaria vector Anopheles gambiae to permethrin decreased following the installation of mosquito nets impregnated with 0.5 g permethrin per square metre in four villages near Kisumu, Kenya. During the first year that permethrin-impregnated bednets and curtains were in place, the exposure time to 50% mortality (LT₅₀) increased 2.5-fold from 13 to 33min, while the LT₅₍₎ for An.gambiae was unchanged in two other villages where no intervention measures were used. Two years after permethrin-impregnated mosquito nets were distributed the LT₅₀s for An.gambiae were 28, 28 and 16min, respectively, in the villages with bednets, curtains and with no such intervention.
Using a colony of An.gambiae derived from females collected in the villages using permethrin-impregnated moquito nets, we lengthened the LT_5n from 28 to 41 min in two generations by exposing all females to permethrin-treated papers for 60 min and rearing offspring of the survivors.
Permethrin-impregnated bednets and curtains are intended to reduce vectorial capacity. Reduced susceptibility to permethrin could counter this beneficial effect.     

Freezing and desiccation tolerance of imbibed canola seed

Depending on the environmental conditions, imbibed seeds survive subzero temperatures either by supercooling or by tolerating freezing-induced desiccation. We investigated what the predominant survival mechanism is in freezing canola ( Brassica napus cv. Quest) and concluded that it depends on the cooling rate. Seeds cooled at 3°C h⁻¹ or faster supercooled, whereas seeds cooled over a 4-day period to −12°C and then cooled at 3°C h⁻¹ to−40°C did not display low temperature exotherms. Both differential thermal analysis and nuclear magnetic resonance (NMR) spectroscopy confirmed that imbibed canola seeds undergo freezing-induced desiccation at slow cooling rates. The freezing tolerance of imbibed canola seed (LT₅₀) was determined by slowly cooling to −12°C for 48 h, followed with cooling at 3°C h⁻¹ to −40°C, or by holding at a constant −6°C (LD₅₀). For both tests, the loss in freezing tolerance of imbibed seeds was a function of time and temperature of imbibition. Freezing tolerance was rapidly lost after radicle emergence. Seeds imbibed in 100 μ M abscisic acid (ABA), particularly at 2°C, lost freezing tolerance at a slower rate compared with water-imbibed seeds. Seeds imbibed in water either at 23°C for 16 h, or 8°C for 6 days, or 2°C for 6 days were not germinable after storage at −6°C for 10 days. Seeds imbibed in ABA at 23°C for 24 h, or 8°C for 8 days, or 2°C for 15 days were highly germinable after 40 days at a constant −6°C. Desiccation injury induced at a high temperature (60°C), as with injury induced by freezing, was found to be a function of imbibition temperature and time.     

INFLUENCE OF TEMPERATURE ON THE MORTALITY OF MYZUS PERSICAE (SULZER) DUE TO THE FUNGAL PATHOGEN ZOOPHTHORA PHALLOIDES BATKO

b
Nymphs of Myzus persicae inoculated with Zoophthora phalloides were killed at each temperature tested in the range 10–22°C. Mortality was around 50% at 12, 15 and 18°C, but was considerably reduced outside this range. Deaths occurred soonest at 18°C (LT₅₀—6.8 days). The low temperature requirement of this fungus limits its potential as a biocontrol agent for aphids in Australia.     

Effect of various gas atmospheres on the growth and survival of Campylobacter jejuni on beef

Pieces of fresh beef were inoculated with three strains of Campylobacter jejuni . The meat was then allocated to three treatments: (a) vacuum packaged, (b) packaged in an atmosphere of 20% CO₂+ 80% N₂, and (c) packaged into sterile Petri dishes in anaerobic cultivation boxes, which were filled with a gas mixture of 5% O₂+ 10% CO₂+ 85% N₂. The packaging material in the first two treatments was PA 80/PE 100–PE 100/PA 80/PE 100. The survival of Campylobacter cells was followed at 37°C, 20°C and 4°C for 48 h, 4 days and 25 days, respectively. At 37°C the counts of two Campylobacter strains increased in each package treatment for 48 h. At 20°C and at 4°C the counts of the same two strains decreased by 1 to 2 log units and 0.5 to 1 log unit, respectively, during storage. The survival of the two strains was about the same in all package treatments. The third strain was the most sensitive of the strains studied. At 37°C its numbers increased only in the optimal gas atmosphere; at 20°C the strain was not detectable after 24 to 48 h storage and at 4°C after 4 days storage. The aerobic plate counts were determined for all samples at the same time as Campylobacter counts. The high indigenous bacterial numbers of the meat samples did not appear to have a great effect on the survival or growth of campylobacters.     

Compensatory growth of juvenile brown flounder Paralichthys olivaceus (Temminck & Schlegel) following thermal manipulation

The compensatory growth of juvenile brown flounder Paralichthys olivaceus (body mass c. 12 g) following different thermal exposure was investigated. Fish were exposed to one of the five temperatures: 8·5 ( T _8·5), 13·0 ( T _13·0), 17·5 ( T _17·5), 22·0 ( T _22·0) and 26·5° C ( T _26·5) for 10 days and fish grew best at 22·0° C. Then the water temperature in all treatments was equably adjusted to 22·0° C over 3 days. At the end of the following 30 days after temperature adjustment, there were no significant differences between body masses of fish in the different treatments (wet body mass at the end of the experiment ranged from 22·13 to 24·56 g). Results indicated that the juvenile P. olivaceus achieved complete compensatory growth. Analysis of the dynamics of the feeding rates and feed conversion efficiencies indicated that compensatory growth of the fish experienced low temperature ( T _8·5, T _13·5 and T _17·5) or high temperature ( T _26·5) exposure was mainly dependent on increasing feed intake (hyperphagia) and possibly by improvement in feed conversion efficiency. The moisture content was not affected by different temperature exposure significantly. The lipid and energy content of juvenile P. olivaceus in T _8·5, however, were significantly lower than other treatment. Results of the current study indicate that a short period of low or high temperature exposure may not affect annual growth, but may affect lipid and energy deposition.     

Natural thermal stress and heat-shock protein expression in Drosophila larvae and pupae

1. Whether Drosophila larvae and pupae naturally experience temperatures that can cause heat damage or death is poorly understood, but bears directly on numerous investigations of the thermal biology and heat-shock response in Drosophila . Accordingly, the temperatures of necrotic fruit, which Drosophila larvae and pupae inhabit, the temperatures of larvae and pupae outside the laboratory, and the levels of the heat-shock protein hsp 70 expressed by larvae in nature were examined.
2. When necrotic fruit was sunlit, internal temperatures rose to levels that can harm indwelling insects. Fruit size and evaporative water loss affected these temperatures. Temperatures of larvae and pupae in the field commonly exceeded 35 °C, with living larvae recorded at >44°C and pupae at >41°C. Natural mortality was evident, presumably because of heat.
3. In the laboratory, these temperatures kill larvae rapidly, with LT₅₀s (time taken for half the sample to be killed) of 30 min at 39 °C, 15 min at 40 °C and 8·5 min at 41 °C. Gradual transfer from 25°C to these temperatures resulted in no lesser mortality than did direct transfer.
4. Hsp 70 levels in lysates of whole larvae were measured by ELISA (enzyme-link immunosorbent assay) with an hsp 70-specific antibody. For larvae within necrotic apples experimentally transferred from shade to sun and within necrotic fruit in situ , hsp 70 levels equalled or exceeded levels detected in parallel laboratory studies of whole larvae or cells in culture.
5. These data provide an ecological context for studies of thermal stress and the heat-shock response in Drosophila that has heretofore been lacking.     

Inhibitory effect of osmotic concentration, potassium and pH on motility of the sperm of the North American burbot Lota lota maculosa

Seminal plasma factors maintaining North American (NA) burbot Lota lota maculosa sperm quiescent were examined. Sperm were diluted into buffered saline solutions of various compositions and motility assessed. After 1 h in these solutions at 10° C, aliquots of the suspension were diluted with tap water and motility again assessed. Dilution of sperm in an incubation solution containing Ca²⁺ in the absence of K⁺ initiated sperm motility resulting in low motility when sperm were subsequently diluted in tap water. Incubation solutions with osmolalities >200 mOsm kg⁻¹ and containing 12·5 mM K⁺ prevented the onset of sperm motility and were associated with maximal sperm motility upon dilution in tap water. Sperm maintained at lower osmolalities exhibited limited motility upon dilution in tap water indicating interdependence between K⁺ and osmolality in maintaining sperm quiescent in the presence of Ca²⁺. Sperm kept in incubation solution at pH values < c. 7·5 for 1 h demonstrated reduced motility when subsequently diluted in tap water. That motility of sperm was pH sensitive was further indicated by CO₂ inhibition of motility. Therefore, NA burbot sperm are probably maintained in an immotile state, yet with potential for motility, by combination of high K⁺, osmolality and possibly pH. The results from this study differ from published information on sperm quiescence in the temporally and geographically distinct Eurasian burbot Lota lota lota .     

Acclimation is beneficial at extreme test temperatures in the Danube crested newt, Triturus dobrogicus (Caudata, Salamandridae)

Despite many studies demonstrating the effect of acclimation on behavioural or physiological traits, considerable debate still exists about the evolutionary significance of this phenomenon. One of the unresolved issues is whether acclimation to warmer temperature is beneficial at treatment or at more extreme test temperatures. To answer this question, we assessed the effect of thermal acclimation on preferred body temperatures ( T _ps), maximum swimming and running speed, and critical thermal maximum ( CT _max) in the Danube crested newt ( Triturus dobrogicus ). Adult newts were kept at 15 °C (control) and 25 °C (treatment) for 8 weeks prior to measurements. We measured T _ps in an aquatic thermal gradient over 24 h, maximum speeds in a linear racetrack at six temperatures (5–33 °C), and CT _max in a continuously heated water bath. T _ps were higher in newts kept at 15 °C than in those kept at 25 °C. The maximum swimming speed did not acclimate. The maximum running speed at 30–33 °C was substantially higher in newts kept at 25 °C than in those kept at 15 °C. CT _max increased with the treatment temperature. Hence, we conclude that the acclimation response to warm temperature is beneficial not at treatment but at more extreme temperatures in newts.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 627–636.