Heat tolerance,temperature acclimation,acute oxidative damage and canalization of haemoglobin expression in <Emphasis Type="Italic">Daphnia</Emphasis>

Daphnia is a widespread freshwater zooplankton species, which is both a classic and emerging new model for research in ecological physiology, ecotoxicology and evolutionary biology of adaptation to novel environments. Heat tolerance in Daphnia is known to depend both upon evolutionary history of a genotype and on individuals’ acclimation to elevated temperature and to correlate with the level of haemoglobin expression. We demonstrate the existence of north–south gradient of heat tolerance in North American D. pulex, which is not associated with any parallel changes in haemoglobin expression. Geographically distinct clones differ in the way their haemoglobin expression changes due to acclimation to a sub-stressful (28°C) temperature, but these changes are not correlated with the latitude of clones’ origin. Likewise, the effect of acclimation to sub-stressful temperature is independent from, and cannot be fully explained by, haemoglobin expression changes during acclimation. The degree of oxidative damage to haemoglobin, measured as the ratio of absorbance at 540:576 nm at the acclimation temperature, is a strong predictor of 28°C-acclimated Daphnia survival during an acute heat exposure. The comparison of haemoglobin expression in resistant and tolerant clones acclimated to different temperatures indicates that tolerant clones exhibit canalization of haemoglobin expression, possessing a high level of haemoglobin even at non-stressful temperatures. We discuss the evolutionary biology of adaptation and acclimation to elevated temperatures in an ecologically important component of freshwater ecosystems in the context of global climate change.     

Dietary microbial levan enhances cellular non-specific immunity and survival of common carp (Cyprinus carpio) juveniles

A preliminary study with a 75days feeding trial was conducted to study the immunomodulatory effect of microbial levan on Cyprinus carpio juveniles. Five purified isonitrogenous and isocaloric diets with graded levels of levan, namely (T(1)) 0.1% levan, (T(2)) 0.2% levan, (T(3)) 0.5% levan, (T(4)) 1.0% levan, and a control group without levan were fed to five groups of fishes in triplicate. The total erythrocyte count and haemoglobin content was significantly (p<0.05) enhanced in the T(3) group, though the total leucocyte counts, total protein and A/G ratio did not show any significant difference (p>0.05) when compared with the control group. The respiratory burst activity (NBT) of blood phagocytes and lysozyme activity was also highest in T(3) group. The relative survival percentage after challenge with Aeromonas hydrophila was highest (100%) in the T(3) group followed by 83.33% and 66.67% in the T(2) and T(4) groups, respectively. This suggests that microbial levan at 0.5% can be used as a dietary immunostimulant for C. carpio juveniles. This is the first report on microbial levan having an immunomodulatory effect on C. carpio.     

The effects of acclimation temperature on pituitary and plasma beta-endorphin in rats at 32.5 degrees C

Endocrine and thermoregulatory responses were studied in male rats exposed to heat (32.5 +/- 0.1 degrees C) from acclimation temperatures of either 24.5 +/- 0.1 degrees C or 29.2 +/- 0.1 degrees C. After 1 hr in the heat, evaporative water loss and tail skin temperature changes in the 24.5 degrees C acclimated rats were greater than in the 29.2 degrees C acclimated rats; both groups displayed similar changes in metabolic rate and rectal temperature. At the respective acclimation temperatures, 29.2 degrees C rats displayed lowered plasma thyroid hormones, elevated beta-endorphin-like immunoreactivity (beta-END-LI) in the plasma, neurointermediate and anterior lobes of the pituitary gland, and no change in plasma corticosterone levels compared to 24.5 degrees C rats. After exposure to 32.5 degrees C for 1 hr, both groups of rats maintained similar plasma corticosterone levels; however, only the 24.5 degrees C group increased plasma thyroxine and beta-END-LI. These data suggest that beta-endorphin may be involved in body temperature regulation during acclimation to elevated environmental temperatures.     

滨海电厂冷却水余热和余氯对中华哲水蚤的影响

依据滨海电厂冷却系统的实际运作情况,模拟研究了滨海电厂冷却水余热和余氯对中华哲水蚤的影响.结果表明：中华哲水蚤的热忍受能力随暴露时间延长而降低,随驯化温度升高而升高.驯化温度为16 ℃～27 ℃的中华哲水蚤在持续受到15、30、45 min热冲击及持续升温暴露24和48 h的致死温度分别为29.9 ℃～31.7 ℃、29.4 ℃～31.0 ℃、28.9 ℃～30.3 ℃和26.9 ℃～28.5 ℃、26.4 ℃～28.0 ℃.当驯化温度升高到一定程度后,其热忍受能力不再上升.电厂冷却水中余氯对中华哲水蚤的毒性随驯化温度升高、升温幅度增大及暴露时间延长而增强.     

Thermoperiodic acclimations enhance cold hardiness of the eggs of the migratory locust

Treatment of thermoperiods that simulate the patterns of natural occurrence is most efficient in enhancing cold hardiness. To examine the effects of different thermoperiods on cold hardiness of eggs in the migratory locust, Locusta migratoria (L.), the survival rates, cryoprotectant levels and three hsps expressions in mid-stage eggs (7-day-old) were measured after the eggs were subjected to three different thermoperiod regimes, : short (2 day), long (10 day), and nature-mimicking thermoperiodic acclimation. The thermoperiodic acclimations resulted in the highest egg survival rates in both the short and the long period acclimation groups in comparison with the groups treated with constant temperatures. The egg survival of nature-mimicking thermoperiod groups was significantly higher than those of constant temperature groups for the same acclimation duration. The survival rate of eggs under single daily thermoperiod was higher than that of multiple daily thermoperiods. The concentration of cryoprotectants (myo-inositol, trehalose, mannitol and sorbitol) and the expression levels of hsp20.5, hsp70, and hsp90 all increased in thermoperiodic acclimation eggs.     

Thermal limits for behavioural function and resistance-adaptation of goldfish,Carassius auratus L.

Summary Goldfish were trained to perform a conditioned avoidance response in a shuttle tank at acclimation temperatures between 10 °C and 35 °C. A high level of success (85–100%) was maintained over a relatively wide range of test temperatures, although outside this range the response was rapidly and reversibly blocked. The upper and lower thermal limits for the avoidance response were determined in goldfish acclimated to temperatures between 10 °C and 35 °C. The absolute thermal limits for the avoidance response in goldfish were approximately 3 °C to 42 °C, but the range for individuals was considerably more restricted. Increased acclimation temperature resulted in higher upper and lower thermal limits and thus constitutes a reasonable resistance adaptation. Over the lower range of acclimation temperatures the upper thermal limit showed greater mobility, whereas over the upper range of acclimation temperatures the lower thermal limits showed greater mobility. Goldfish acclimated to 5 °C and 38.5 °C exhibited very reduced % success at their respective acclimation temperatures even though they showed high % success when the same individuals were previously acclimated to less stressful temperatures. These extreme acclimation temperatures probably represent the absolute limits for chronic exposure.     

Haematological values during normal reproduction of the maternal and the fetal rabbit

1. Haematological values of non-pregnant/non-lactating, pregnant as well as lactating rabbits and 28-day-old fetuses were measured. 2. The haemoglobin content in does decreased during the observed periods from 122 +/- 8 g/l to 100 +/- 11 g/l. In 28-day-old fetuses it was 85 +/- 0 g/l. 3. The erythrocyte count in 28-day-old fetuses was 2.4 X 10(12)/l. In the does, the erythrocyte count was 5.2 X 10(12)/l in week 4 of gestation. The erythrocyte volume in fetuses was about 45% higher than that of the doe. 4. In fetuses the leucocyte count was approximately one ninth that of the mother in week 4 of gestation (0.41 +/- 0.08 X 10(9)/l vs 3.8 +/- 0.4 X 10(9)/l).     

Sub-lethal temperature thresholds indicate acclimation and physiological limits in brook trout Salvelinus fontinalis

The upper thermal tolerance of brook trout Salvelinus fontinalis was estimated using critical thermal maxima (CT_max) experiments on fish acclimated to temperatures that span the species' thermal range (5–25°C). The CT_max increased with acclimation temperature but plateaued in fish acclimated to 20, 23 and 25°C. Plasma lactate was highest, and the hepato-somatic index (I_H) was lowest at 23 and 25°C, which suggests additional metabolic costs at those acclimation temperatures. The results suggest that there is a sub-lethal threshold between 20 and 23°C, beyond which the fish experience reduced physiological performance.     

The onset temperature of the heat-shock response and whole-organism thermal tolerance are tightly correlated in both laboratory-acclimated and field-acclimatized tidepool sculpins (Oligocottus maculosus)

We examined the relationship between thermal tolerance, measured as critical thermal maximum (CT(max)), and aspects of the heat-shock response in tidepool sculpins (Oligocottus maculosus) acclimated to constant laboratory temperatures or acclimatized to field conditions. The CT(max) of fish laboratory acclimated to 6°, 13°, and 20°C were 27.6° ± 0.1°C, 29.5° ± 0.1°C, and 30.8° ± 0.1°C, respectively, increasing linearly by 0.2°C for each 1°C increase in acclimation temperature. The CT(max) of field-acclimatized fish from the low intertidal (29.9° ± 0.1°C) was significantly lower than that of fish from the mid- (30.5° ± 0.1°C) and high (30.4° ± 0.1°C) intertidal. CT(max) and the onset temperature of hsp70 induction in gill (T(on)) were highly correlated in both laboratory-acclimated and field-acclimatized sculpins, with T(on) occurring at 2°C below CT(max) in all cases. However, there was no consistent relationship between CT(max) and the maximum levels of gill hsp70 mRNA. Predicted "acclimation" temperature (15.9° ± 0.3°C) and mean habitat temperature (15.9° ± 1.6°C) were similar for sculpins from low intertidal pools, but this relationship was not apparent in mid- and high intertidal fish. Mark-recapture experiments indicated that approximately 80% of fish from low intertidal pools were residents of that pool, but residency rates were less than 50% in mid- and high intertidal pools, which may explain the lack of correlation between CT(max) and habitat variables in these groups. These data indicate that gill hsp70 T(on) and CT(max) are highly correlated indicators of the thermal performance of tidepool sculpins in both laboratory and field settings.     

Changes in the blood of Wistar rats in acute poisoning with ethanol and acetaldehyde

1. The purpose of the study was to investigate the effect of ethanol and acetaldehyde on the erythrocyte and leucocyte system of Wistar rats. 2. Administration of the ethanol or acetaldehyde caused a considerable drop in the number of erythrocytes, haemoglobin level and haematocrit value in rats. 3. The mean erythrocyte volume was smaller after only 0.5 hr of exposure to ethyl alcohol. 4. The solutions used caused changes in the leucocyte system expressed in distinct neutrophilic leucocytosis. 5. Changes in the leucogram were reflected in the increase in the leucocyte index. 6. The degree of intensity of changes in both the erythrocyte and leucocyte system point to the greater toxicity of ethyl alcohol intoxication than is the case of acetaldehyde in a toxically corresponding dose (i.e. 0.5 and 5 g/kg body wt respectively).     

Cold-induced ependymin expression in zebrafish and carp brain: implications for cold acclimation.

Cold acclimation has been suggested to be mediated by alternations in the gene expression pattern in the cold-adapted fish. To investigate the mechanism of cold acclimation in fish brain at the molecular level, relevant subsets of differentially expressed genes of interest were identified and cloned by the PCR-based subtraction suppression hybridization. Characterization of the selected cold-induced cDNA clones revealed one encoding ependymin. This gene was shown to be brain-specific. The expression of ependymin was induced by a temperature shift from 25 degrees C to 6 degrees C in Cyprinus carpio or 12 degrees C in Danio rerio. Activation of ependymin was detected 2 h after cold exposure and peaked at more than 10-fold at 12 h. This peak level remains unchanged until the temperature returns to 25 degrees C. Although the amount of soluble ependymin protein in brain was not changed by cold treatment, its level in the fibrous insoluble polymers increased 2-fold after exposure to low temperature. These findings indicate that the increase in ependymin expression is an early event that may play an important role in the cold acclimation of fish.     

Field and laboratory investigations of the thermal influence on tissue-specific Hsp70 levels in common carp (Cyprinus carpio).

Thermal discharge from power stations can affect normal environmental conditions and change in heat shock proteins expression of native fish with increasing temperature. In this study, we investigated levels of Hsp70 in the heart, kidney, brain and gill of the common carp Cyprinus carpio both in long-term heat discharge environment and after 24 h acute heat shock exposure. In laboratory exposure experiments, fish acclimated at 10 degrees C were exposed to various elevated temperatures (20, 24 and 28 degrees C). Hsp70 concentrations were determined in tissues by Western blotting analysis after one dimensional SDS-PAGE separation. In the field study, the level of Hsp70 in the gill of the carp remained at control values, and Hsp70 expression in the heart, kidney and brain underwent a 2.8 to 3.7-fold increase. A lower thermal sensitivity of the Hsp70 response of the brain, compared with the heart, kidney and gill, was observed in the laboratory experiments. Our data show that these tissues had different levels of Hsp70 responses to thermal influence both in acute exposure and long-term acclimation. The pattern of tissue Hsp70 expression may have a close relationship with the thermal tolerance of the carp and allows the fish to survive long-term thermal pollution.     

The influence of haemoglobin on behavioural thermoregulation and oxygen consumption in Daphnia carinata

When placed in a temperature gradient, most ectotherms have a strict thermal preference that is lowered on exposure to hypoxia. Branchiopods, small aquatic crustaceans, are known to synthesise haemoglobin (Hb) when exposed to hypoxia; hypoxia can occur diurnally and seasonally in ponds. The effect of Hb on behavioural thermoregulation in the branchiopod Daphnia carinata following exposure to both normoxia and hypoxia was examined. Control animals raised in normoxia (Po2=150 mmHg, [Hb]=0.026+/-0.007 mg g dry wt-1) and Hb-rich animals raised in hypoxia (Po2=70 mmHg, [Hb]=0.080+/-0.017 mg g dry wt-1) were placed (N=30) in a tube (length=500 mm, diameter=8 mm) filled with pond water. In the absence of a thermal gradient, control and Hb-rich animals in normoxic water were uniformly distributed along the tube. The presence of a thermal gradient (13 degrees -28 degrees C) elicited clustering at a preferred temperature, T approximately 23 degrees C for both groups. Exposure to hypoxic water in a thermal gradient resulted in a behavioural shift: T approximately 16 degrees C for controls and T approximately 19 degrees C for Hb-rich animals. Measurements of oxygen consumption (V&d2;o2) at fixed temperatures revealed that Hb is associated with a metabolic acclimation to hypoxia.     

Regulation of the cardiorespiratory system of common carp (Cyprinus carpio) during severe hypoxia at three seasonal acclimation temperatures

Little is known of the cardiorespiratory control mechanisms utilized by hypoxia-tolerant teleost fish to tolerate prolonged periods (h) of near anoxic exposure. Here, we report on the cardiorespiratory control mechanisms of the common carp Cyprinus carpio L. during normoxia and prolonged, severe hypoxic (<0.3 mg O(2) L(-1)) exposure at acclimation temperatures of 5 degrees C, 10 degrees C, and 15 degrees C. Through serial intra-arterial injections of alpha - and beta -adrenergic, cholinergic, and purinergic antagonists while measuring cardiac output (Q), heart rate (f(H)), ventral aortic blood pressure, and respiration rate, we established that autonomic cardiovascular and respiratory control was preserved during severe hypoxia at all three acclimation temperatures and contributed to downregulation of cardiorespiratory activity. Specifically, inhibitory cholinergic tone mediated up to 76% reductions in f(H) and Q during hypoxia, whereas the accompanying arterial hypotension was attenuated by an upregulation of an alpha -adrenergically mediated peripheral vasoconstriction. Despite the overall cardiac downregulation, a large, stimulatory cardiac beta -adrenergic tone was present during prolonged, severe hypoxia, possibly to protect the heart from attendant acidotic conditions. Purinergic blockade, following alpha -adrenergic and cholinergic antagonists, showed that the hypoxic ventilatory depression, which reversed the 2.3- to 7.7-fold increases in respiration rate that occurred with the onset of hypoxia, was a result of purinergic inhibition at all three acclimation temperatures. In contrast, purinergic inhibition of cardiac activity during hypoxia might be important only at 5 degrees C. Finally, given that cardiac power output was reduced 72%-87% during prolonged, severe hypoxia and that glycolysis yields approximately 94% less ATP per mole glucose than oxidative phosphorylation, it seems unlikely that the common carp sufficiently reduces its cardiac energy demand to a level to preclude activation of a partial Pasteur effect. This means that glycogen stores will be used and waste products will accumulate at faster rates, a finding that may help explain why the common carp cannot tolerate such extended periods of severe hypoxia (weeks to months) at cold acclimation temperatures as the freshwater turtle, which is able to reduce its cardiac energy demand to a level that does not require a Pasteur effect and also blunts autonomic cardiovascular control.     

Curcuma longa as feed additive in broiler birds and its patho-physiological effects

Broiler birds (Vencob chicken of 3 days old) when given feed mixed with powdered rhizome of Curcuma longa (CL; @ 1 g/kg) for 42 days of age, showed significant decrease in serum uric acid and albumin as compared to control, whereas significant increase was recorded in the level of serum total protein and globulin. Level of serum glucose, alkaline phosphatase, aspartate amino transferase and calcium showed no significant variation between the two groups. Micronutrient assay revealed significantly higher level of manganese, zinc, iron and copper in treated group as compared to control group. HA/HI test revealed better humoral response against RD vaccine in CL administered birds. Haematological study showed significantly higher haemoglobin and absolute neutrophil count in treated group. Addition of CL as feed additive also resulted in better growth rate, feed consumption and F:C efficiency in the treated birds. Thus, it could be concluded that powdered CL might be a useful feed additive, since it enhanced the F:C efficiency and had nephroprotective properties.     

Photosynthetic and respiratory acclimation and growth response of Antarctic vascular plants to contrasting temperature regimes

Air temperatures have risen over the past 50 yr along the Antarctic Peninsula, and it is unclear what impact this is having on Antarctic plants. We examined the growth response of the Antarctic vascular plants Colobanthus quitensis (Caryophyllaceae) and Deschampsia antarctica (Poaceae) to temperature and also assessed their ability for thermal acclimation, in terms of whole-canopy net photosynthesis (P(n)) and dark respiration (R(d)), by growing plants for 90 d under three contrasting temperature regimes: 7°C day/7°C night, 12°C day/7°C night, and 20°C day/7°C night (18 h/6 h). These daytime temperatures represent suboptimal (7°C), near-optimal (12°C), and supraoptimal (20°C) temperatures for P(n) based on field measurements at the collection site near Palmer Station along the west coast of the Antarctic Peninsula. Plants of both species grown at a daytime temperature of 20°C had greater RGR (relative growth rate) and produced 2.2-3.3 times as much total biomass as plants grown at daytime temperatures of 12° or 7°C. Plants grown at 20°C also produced 2.0-4.1 times as many leaves, 3.4-5.5 times as much total leaf area, and had 1.5-1.6 times the LAR (leaf area ratio; leaf area:total biomass) and 1.1-1.4 times the LMR (leaf mass ratio; leaf mass:total biomass) of plants grown at 12° or 7°C. Greater RGR and biomass production at 20°C appeared primarily due to greater biomass allocation to leaf production in these plants. Rates of P(n) (leaf-area basis), when measured at their respective daytime growth temperatures, were highest in plants grown at 12°C, and rates of plants grown at 20°C were only 58 (C. quitensis) or 64% (D. antarctica) of the rates in plants grown at 12°C. Thus, lower P(n) per leaf area in plants grown at 20°C was more than offset by much greater leaf-area production. Rates of whole-canopy P(n) (per plant), when measured at their respective daytime growth temperatures, were highest in plants grown at 20°C, and appeared well correlated with differences in RGR and total biomass among treatments. Colobanthus quitensis exhibited only a slight ability for relative acclimation of P(n) (leaf-area basis) as the optimal temperature for P(n) increased from 8.4° to 10.3° to 11.5°C as daytime growth temperatures increased from 7° to 12° to 20°C. There was no evidence for relative acclimation of P(n) in D. antarctica, as plants grown at all three temperature regimes had a similar optimal temperature (10°C) for P(n). There was no evidence for absolute acclimation of P(n) in either species, as rates of P(n) in plants grown at a daytime temperature of 12°C were higher than those of plants grown at daytime temperatures of 7° or 20°C, when measured at their respective growth temperatures. The poor ability for photosynthetic acclimation in these species may be associated with the relatively stable maritime temperature regime during the growing season along the Peninsula. In contrast to P(n), both species exhibited full acclimation of R(d), and rates of R(d) on a leaf-area basis were similar among treatments when measured at their respective daytime growth temperature. Our results suggest that in the absence of interspecific competition, continued warming along the Peninsula will lead to improved vegetative growth of these species due to (1) greater biomass allocation to leaf-area production (as opposed to improved rates of P(n) per leaf area) and (2) their ability to acclimate R(d), such that respiratory losses per leaf area do not increase under higher temperature regimes.