Effect of acclimation temperature on thermoregulatory behaviour,thermal tolerance and respiratory metabolism of Lutjanus guttatus and the response of heat shock protein 70 (Hsp70) and lactate dehydrogenase (Ldh‐a) genes

This study evaluated the effect of different environmental temperatures in the physiology of Lutjanus guttatus juveniles by analysing their thermoregulatory behaviour, thermal tolerance, oxygen consumption rates and thermal metabolic scope. Jointly, the effect of acclimation and critical temperatures on heat shock protein 70 (Hsp70) and lactate dehydrogenase (Ldh‐a) gene expressions were also analysed using acclimation temperatures of 20, 23, 26, 29 and 32°C. The results showed that the final preferred temperature in juvenile snapper was 26°C with a thermal window of 336.5°C², which was related to an optimal temperature for their physiology determined by the routine metabolic rate and thermal metabolic scope. At temperatures from 20 to 26°C, the routine metabolic rate and Hsp70 and Ldh‐a genes had the lowest values related to a basal expression level. At acclimation temperatures from 29 to 32°C and after critical thermal maximum (CT_max) limit, the relative expression of Hsp70 and Ldh‐a genes increased significantly, but the main response at CT_max was the upregulation of Hsp70 gene.     

Effects of temperature on growth of juvenile blackfoot abalone, Haliotis iris Gmelin

The effects of temperature on growth and survival of juvenile blackfoot abalone, Haliotis iris, were investigated. Animals of 10, 30 or 60 mm initial shell length were exposed to ambient (6–10°C), 14, 18, 22 and 26°C for 112 days in a flow‐through culture system. Maximum growth occurred at 22°C for the 10 and 30 mm size classes and at 18°C for the 60 mm size class. Regression analysis identified the optimal temperature for growth (T_optG) at around 21°C for the 10 and 30 mm size classes and at 17–18°C for the largest size class. In a second experiment, the critical thermal maximum of H. iris was determined as a measure of thermal tolerance. Abalone were subjected to increasing water temperatures at a rate of 2°C h^?1 until they detached from the substrate. Abalone of 10 mm displayed greater thermal tolerance than abalone of 30 and 60 mm in length. CT₅₀ temperatures were 28.8, 27.7 and 27.8°C, yielding deduced T_optG values of 19.7, 18.3 and 18.4°C for the 10, 30 and 60 mm size classes respectively. The size‐dependent nature of the relationship between growth and temperature could be capitalized upon in recirculating aquaculture systems.     

Effects of air exposure on the lysosomal membrane stability of haemocytes in blacklip abalone, Haliotis rubra (Leach)

The neutral red retention (NRR) assay was used to evaluate the effects of air exposure on lysosomal membrane integrity in the haemolymph of blacklip abalone, Haliotis rubra, and its subsequent recovery in water. After acclimation in 16°C water for 7 days, abalone were exposed to an air temperature of 7, 16 or 23°C for 12 h in the air exposure experiment or to these three air temperatures, e.g., for 12, 24 or 36 h, followed by re‐immersion in 16°C water in the lysosomal membrane stability recovery experiment. Statistical analyses of the air exposure experiment showed that when abalone were exposed to different air temperatures (7, 16 or 23°C), the lysosomal membrane stability was significantly affected by the air temperature, the exposure duration and their interaction. Air temperature similar to the acclimation temperature had a significantly lower impact on the lysosomal membrane stability within the initial 4.5 h in comparison with the other two temperatures in the same period. The lysosomal membrane stability recovery experiment showed that after air exposure durations of 12, 24 or 36 h, the re‐stabilization of the lysosomal membrane was faster in the animals exposed to lower temperatures than those exposed to higher temperatures. The recovery of the lysosomal membrane stability in abalone exposed to lower 7°C air temperature was not significantly affected (F_{2, 66}=0.251, P=0.779) by the exposure durations (12, 24 and 36 h) used in this study. Alternatively, the lysosomal membrane stability in abalone exposed to higher air temperatures of 16 or 23°C recovered at a faster rate when subjected to shorter durations of air exposure (F_{2, 66}=3.663, P=0.031 and F_{1, 44}=17.057, P<0.001 for 16 and 23°C respectively).     

皱纹盘鲍南北方群体的高温应激和生长差异

为研究皱纹盘鲍(Haliotis discus hannai Ino)经过杂交和南方养殖后,南方群体(杂交群体)是否与北方群体(自交群体)在适温性上是否产生了差异,从而是否更加适应南方高温环境,本研究开展了皱纹盘鲍南北方群体的高温应激和南方地区养殖实验。高温应激实验中:经过30℃应激后,南方群体恢复期所有取样组织中HSP70在0 h的表达量都显著高于北方群体(P0.05),HSP90在外套膜和肝胰脏中的表达量也高于北方群体(鳃中的表达量略低但差异不显著);经过33℃应激后,南方群体HSP70和HSP90在足、外套膜和鳃0 h表达量都显著高于北方群体(P0.05)。养殖实验中:从12月开始至翌年2月水温不超过17.11℃,为北方群体的快速生长期,生长快于南方群体;从3月开始,水温逐渐超过20℃,北方群体生长慢于南方群体,同时北方群体开始出现死亡现象,7月北方群体大量死亡,存活数量不足30粒。综合HSP70、HSP90的高温应激表达和养殖实验结果,可以得出:皱纹盘鲍经过杂交和南方累代养殖后,适温上限有所提高,已与北方群体产生较大差异,可适应较高的水温;30℃水温刺激下南北方群体的HSP70的相对表达量与对高温的耐受性具有正相关性,可以在鲍的抗高温选育种中进行辅助筛选。     

Growth and feeding in juvenile triploid and diploid blacklip abalone, Haliotis rubra (Leach, 1814), at two temperatures

Growth and feeding of juvenile triploid and diploid blacklip abalone Haliotis rubra (Leach, 1814) were investigated at two temperatures of 17 and 21 °C over a 50‐day period. There were no differences in growth between triploid and diploid abalone as measured by shell length and body weight. Both triploid and diploid abalone increased in length but not in weight at 21 °C. Condition indices were similar for triploid abalone maintained at both temperatures; however, those for diploid abalone were significantly higher at 17 °C than at 21 °C. Food intake was significantly greater yet feed conversion efficiency was significantly lower in triploid than in diploid abalone. Both the feeding variables were independent of temperature. On average, diploid abalone were able to convert 1 g of dry food ingested to 0.58 g of body weight, but triploid abalone only 0.44 g.     

The physiological tolerance of the grey carpet shark (<Emphasis Type="Italic">Chiloscyllium punctatum</Emphasis>) and the epaulette shark (<Emphasis Type="Italic">Hemiscyllium ocellatum</Emphasis>) to anoxic exposure at three seasonal temperatures

The epaulette shark (Hemiscyllium ocellatum) and the grey carpet shark (Chiloscyllium punctatum) are commonly found in periodically hypoxic environments. The ecophysiological time available for these animals to safely exploit these niches during different seasonal temperatures was examined. The time to loss of righting reflex (T _LRR) was examined in response to an open ended anoxic challenge at three seasonal temperatures (23, 25 and 27°C). Ventilation rates were measured in an open ended anoxic challenge at 23°C and during 1.5 h of anoxia followed by 2 h of re-oxygenation at 23 and 25°C. The mean T _LRR of epaulette and grey carpet sharks was inversely proportional to temperature. The T _LRR was similar between species at 23°C; however, grey carpet sharks had significantly reduced T _LRR at higher temperatures. During the standardised anoxic challenge, epaulette sharks entered into ventilatory depression significantly earlier at 25°C. During re-oxygenation, epaulette sharks exposed to anoxia at 23°C had no significant increase in ventilation rates. However, after anoxic challenge and re-oxygenation at 25°C, epaulette sharks showed a significant increase in ventilation rates during re-oxygenation. Grey carpet sharks displayed no evidence of ventilatory depression during anoxia. However, during re-oxygenation, grey carpet sharks had significantly elevated ventilation rates above pre-experimental levels and control animals. These data demonstrate that the anoxia tolerance times of both species were temperature dependent, with a significant reduction in the T _LRR occurring at higher temperatures. Epaulette sharks had a significantly greater T _LRR at higher temperatures than grey carpet sharks, which did not enter into a ventilatory depression.     

Effect of thermal stress on Hsp70 gene expression and female reproductive performance of giant freshwater prawn,Macrobrachium rosenbergii

Using Hsp70 as a biomarker, thermal stress impinges on reproductive organs, ovary and hepatopancreas were being analyzed by determining the expression of Hsp70 mRNA inside the organs after the adult inter‐molt females were subjected to thermal treatment at 35, 30 and 28°C (Control). Results showed the expression of Hsp70 mRNA under thermal treatment of 35°C after 2 hr recovery in ovary were upregulated at 2, 4, 6, 12, 24 hr and 30 days compared to control whereas in hepatopancreas under similar treatment, the expression of Hsp70 mRNA were significantly higher than control at 6, 24 hr and 30 days. Frequency of reproductive molt at 35°C showed the ovary of females were failed to develop and only entered common molt along three consecutive molt cycles. For 30°C thermal treatment, the expression of Hsp70 mRNA was significantly higher than control after 2 hr recovery but returned to normal afterwards until 30 days’ thermal treatment. Maternal heat shock for 2 hr at 35°C were found to give significantly lower frequency of reproductive molt and longer duration of ovarian development and incubation period whereas maternal heat shock for 2 hr at 30°C gave lower frequency of reproductive molt, slower development of embryo and lower hatching success compared to untreated control. This study suggests that short and long‐term thermal stress at 30 and 35°C were found to affect the induction of Hsp70 mRNA in reproductive organs of Macrobrachium rosenbergii and also influence their reproductive performance.     

Effects of temperature and dietary protein on gene expression of Hsp70 and Wap65 and immunity of juvenile mirror carp (Cyprinus carpio)

Juvenile mirror carp were fed diets containing 303.4, 321.7, 341.2, 361.0 and 379.1 g kg^?1 proteins, respectively, and reared at different water temperatures (18, 23 and 28°C) for 60 days. Gene expression of heat shock protein gene (Hsp70) and the warm temperature acclimation‐related 65 kDa protein gene (Wap65), immunity and antioxidant status in the carp were investigated. Results indicated that the contents of serum complement 3 (C3), complement 4 (C4) and immunoglobulin M (IgM), as well as activities of liver superoxide dismutase (SOD) and lysozyme (LSZ) were significantly enhanced with increasing dietary protein (P < 0.05), while content of malondiadehyde (MDA) decreased. Gene expression level of Wap65 in the liver significantly increased with dietary protein, while gene expression of Hsp70 decreased. The contents of C3, C4 and IgM, the activities of SOD and LSZ and gene expression level of Wap65 in the liver significantly increased with temperature. These results suggest that: Serum immune parameter, antioxidant enzymes and Hsp70 and Wap65 expression interact in fish to improve ability to adapt to the environment; and the optimal conditions for the immunity of carp are 348.1?354.5 g kg^?1 protein at 18°C, 352.3?364.9 g kg^?1 at 23°C and 360.2?364.3 g kg^?1 at 28°C, and the optimum temperature for carp is 23°C.     

Interactions between temperature and size on the growth,size heterogeneity,mortality and cannibalism in cultured larvae and juveniles of the Asian catfish,Pangasianodon hypophthalmus (Sauvage)

Temperature effects vary throughout the ontogeny, but are proportionally more variable during the early life stages than in older fish. The larvae of a few species have been studied but contrasting trajectories were observed, thereby making it difficult to predict how temperature impacts on the growth, survival and size heterogeneity in a particular species. This study examined these interactions in young [0.9 mg–1 g wet mass (WM)] Pangasianodon hypophthalmus, one of the most extensively cultured tropical fish. Fish were raised at five temperatures from 23 to 33 °C in a recirculated water system, fed in excess with a high‐energy feed (Artemia then a formulated feed, >50% protein), and examined at 1‐day (first feeding days) or 4‐day intervals (older fish). The temperature that produced the fastest growth (T°_opt) was 31 °C at the start of exogenous feeding (0.9 mg); it increased to 32.7 °C at 8 mg then decreased by 0.7 °C for each 10‐fold increase of WM. Size heterogeneity was lower and survival was higher (70–85% from 0.9 mg to 1 g) when the temperature was close to T°_opt. Comparisons with other species suggest that the slope of the relationship between T°_opt and fish size has a strong latitudinal component, and may also reflect the fish sensitivity to oxygen.     

Effects of heat-shock selection during pelagic stages on thermal sensitivity of juvenile sea cucumber, <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka

The effects of heat-shock during pelagic stages on growth and survival of juvenile sea cucumber Apostichopus japonicus were investigated to test the possibility to acquire high thermal tolerance individuals after the heat-shock selection. Larvae at the stages of blastula, gastrula, and auricularia were heat-shocked at selected temperatures (21.5, control; 26, 28, and 30°C) for 45 min and returned to 21.5°C for continuous rearing. There was a significant difference in thermal sensitivity among different developmental stages, and thermal tolerance of the larvae was correlated with the expression of heat shock protein 70 (Hsp70). Juveniles after the heat-shock selection at pelagic stages showed higher induced thermotolerance than those without heat-shock. Therefore, heat-shock application at early pelagic development stages is potentially a useful way to select high-thermotolerance variety of the sea cucumbers.     

Sporophytic photosynthesis and gametophytic growth of the kelp Ecklonia stolonifera affected by ocean acidification and warming

Juvenile sporophytes and gametophytes of Ecklonia stolonifera were incubated in combinations of three pCO₂ levels (360, 720 and 980 ppmv) and two temperatures (10 and 15°C for sporophytes; 15 and 20°C for gametophytes) to examine potential effects of climate change on photosynthesis and growth. Sporophytes had significantly higher maximum quantum yields (F_v/F_m) and maximum relative electron transport rates (rETR_max) at 720 ppmv than 360 and 980 ppmv. Also, these parameters were significantly lower at higher temperature of 15°C than at 10°C. Growth of female gametophytes was maximal at 360 ppmv rather than enriched pCO₂ levels. Female gametophytes had significantly lower growth at higher temperature of 20°C than at 15°C. These results indicate effects of elevated pCO₂ varied between generations: stimulating sporophytic photosynthesis and inhibiting gametophytic growth. Ocean acidification and warming would constitute a grave threat to seedling cultivation of E. stolonifera caused by growth inhibition of gametophytes at high pCO₂ levels and temperatures.     

Effect of temperature on excess post-exercise oxygen consumption in juvenile southern catfish (<Emphasis Type="Italic">Silurus meridionalis</Emphasis> Chen) following exhaustive exercise

The effects of temperature on resting oxygen consumption rate (MO_2rest) and excess post-exercise oxygen consumption (EPOC) after exhaustive exercise (chasing) were measured in juvenile southern catfish (Silurus meridionalis) (8.40 ± 0.30 g, n = 40) to test whether temperature has a significant influence on MO_2rest, maximum post-exercise oxygen consumption rate (MO_2peak) and EPOC and to investigate how metabolic scope (MS: MO_2peak − MO_2rest) varies with acclimation temperature. The MO_2rest increased from 64.7 (10°C) to 160.3 mg O₂ h⁻¹ kg⁻¹ (25°C) (P < 0.05) and reached a plateau between 25 and 30°C. The post-exercise MO₂ in all temperature groups increased immediately to the peak values and then decreased slowly to a steady state that was higher than the pre-exercise MO₂. The MO_2peak did not significantly differ among the 20, 25 and 30°C groups, though these values were much higher than those of the lower temperature groups (10 and 15°C) (P < 0.05). The duration of EPOC varied from 32.9 min at 10°C to 345 min at 20°C, depending on the acclimation temperatures. The MS values of the lower temperature groups (10 and 15°C) were significantly smaller than those of the higher temperature groups (20, 25 and 30°C) (P < 0.05). The magnitude of EPOC varied ninefold among all of the temperature groups and was the largest for the 20°C temperature group (about 422.4 mg O₂ kg⁻¹). These results suggested that (1) the acclimation temperature had a significant effect on maintenance metabolism (as indicated by MO_2rest) and the post-exercise metabolic recovery process (as indicated by MO_2peak, duration and magnitude of EPOC), and (2) the change of the MS as a function of acclimation temperature in juvenile southern catfish might be related to their high degree of physiological flexibility, which allows them to adapt to changes in environmental conditions in their habitat in the Yangtze River and the Jialing River.     

Physiological and biochemical responses of Zhikong scallop,Chlamys farreri,to different thermal stressors

Water temperature is a significant environmental stressor that affects physiology and biochemical activities of bivalves. Here, temporal variations in physiological and biochemical parameters of Zhikong scallop, Chlamys farreri, under three water temperature treatments were investigated. For the first treatment, scallops were transferred from rearing temperature (15°C, control temperature) to 5, 10, 20 and 25°C, respectively. The second group of scallops was gradually acclimated to above experimental temperatures at a rate of 1–2°C day^?1. The third group was transferred directly between 15 and 7°C every 12 h and for six times. Results showed that significantly higher oxygen consumption and ammonia‐N excretion together with a significant lower ingestion rate was observed for the acute temperature change treatment compared to those in the equivalent gradual temperature variation treatment (P < 0.05). In acute temperature change treatment, hepatopancreas antioxidant enzyme activities, that is, superoxide dismutase (SOD) and catalase (CAT), immune enzyme activities, that is, acid phosphatase (ACP) and lysozyme (LSZ), and heat‐shock protein 70 gene (Hsp70) expression levels of scallops increased substantially within 48 h. Significant increases in SOD, CAT, ACP and LSZ activities, and malondialdehyde content occurred under exposure to fluctuating temperatures (P < 0.01). Gill and hepatopancreatic Hsp70 expression levels also increased significantly in response to water temperature fluctuations (P < 0.05). The study provides basic knowledge about thermal stress in Zhikong scallop and may contribute to the management of scallop mortalities.     

Effect of Hybrid Abalone,Haliotis discus hannai × Haliotis discus discus,Cultivation on the Carbon Cycle: Carbon Source/Sink

Respiration, calcification, and bio‐deposition of hybrid abalone, Haliotis discus hannai × Haliotis discus discus, fed on different foodstuffs have been measured to evaluate the effect of hybrid abalone culture on carbon source/sink in coastal areas. Fed with Laminaria japonica, Undaria pinnatifida, Gracilaria lemaneiformis, U. pinnatifida, and Ulva pertusa, alternated mutually, the carbon bio‐deposition rate of hybrid abalone was 24.29 ± 6.39, 65.40 ± 10.55, 21.48 ± 5.99, and 29.28 ± 6.47 µg/g/h, respectively. Hybrid abalone fed on U. pinnatifida had a higher carbon bio‐deposition rate compared to that fed on other foodstuff (P < 5%). Rate of CO₂ released by respiration of hybrid abalone fed on the experimental foodstuff was 24.53 ± 8.57, 32.73 ± 7.99, 29.31 ± 6.39, and 33.67 ± 12.37 µg/g/h, respectively. Results indicated that calcification presented less relationship with body weight type of the foodstuff. The rate of CO₂ released by calcification into seawater and atmosphere was 2.77 ± 1.89 and 6.53 ± 3.36 µg/g/h, respectively. The total rate of CO₂ released because of bio‐deposition, respiration, and calcification processes was 16.19 ± 4.67 µg/g/h, while the total rate of carbon sequestered in shells and tissues was 8.94 ± 2.07 µg/g/h. The study revealed that hybrid abalone culture is a source of CO₂.     

Thermal preference and tolerance of green abalone Haliotis fulgens (Philippi, 1845) and pink abalone Haliotis corrugata (Gray, 1828)

The thermoregulatory behaviour of green abalone Haliotis fulgens and pink abalone H. corrugata was investigated. Haliotis fulgens juveniles ranging in wet weight from 3.0 to 3.3 g and from 28.7 to 30.5 mm shell length and of H. corrugata 2.0 g and 25.7 mm in shell length were exposed to 19°C for 30 days in a flow‐through water system. Temperature preference was determined in a horizontal thermal gradient and was found to be 25.4°C for green abalone and 25.0°C for pink abalone. Displacement velocity was 4.3 cm h⁻¹ for H. fulgens and 12.8 cm h⁻¹ for H. corrugata. The optimum temperature for growth calculated for both abalone species was 24.6 and 24.5°C respectively. The critical thermal maxima (CTMax) of H. fulgens and H. corrugata were determined as a measure of thermal tolerance. Abalones were subjected to increasing water temperatures at a rate of 1°C on 30 min until they detached from the substrate. The CTMax at 50% were 33.6 and 32.0°C for green and pink abalone respectively. The results are discussed in relation to site selection and commercial rearing.     

Comparison of the growth performance of offspring from cultured versus wild populations of arctic charr,Salvelinus alpinus (L.), kept at three different temperatures

The effects of three different temperatures on the growth and maturation of the offspring of cultured versus wild populations of juvenile arctic charr (Salvelinus alpinus L.) were investigated. The fish (start weight 17 g) were reared for 5 months at constant temperatures of 12, 14 and 16°C under a continuous light regime (LD24:0). Growth performance was significantly influenced by both temperature and source population. The offspring from the farmed fish displayed significantly higher mean weights at all temperatures compared with the offspring from the wild fish. The results indicate that the optimal temperature for growth (T_opt) decreases with increasing fish size in the offspring of both cultivated and wild fish. Significant differences in length–weight relationship were found, with the offspring of wild fish displaying higher condition factor throughout the experiment. The results demonstrate that the offspring from multiple generation farmed population of arctic charr are better adapted to high temperatures compared with the offspring (F₁ generation) from a wild population of arctic charr, and the former are also able to maintain growth at higher temperatures. There was a significant difference in maturation between the two populations, as the offspring from cultured fish displayed significantly lower level of maturation: 0%, 4% and 2% mature compared with 24%, 40% and 42% in the offspring from wild fish at temperatures of 12, 14 and 16°C respectively. The offspring from farmed strains of arctic charr thus appear to be the most suitable population for aquaculture.     

Optimum temperature for growth of Australian abalone: preferred temperature and critical thermal maximum for blacklip abalone, Haliotis rubra (Leach), and greenlip abalone, Haliotis laevigata (Leach)

The preferred temperature and critical thermal maximum of Australian blacklip abalone, Haliotis rubra (Leach), and greenlip abalone, Haliotis laevigata (Leach), were found to differ only slightly; the blacklip abalone exhibited lower temperature tolerance and preference, as expected from its habitat distribution. Preferred temperatures were 16.9 and 18.9°C, and 50% critical thermal maxima were 26.9 and 2 7.5°C for blacklip and greenlip abalone, respectively. The optimum temperatures for growth calculated from each of these indices and averaged were 17.0 and 18.3°C, respectively.     

Hybridization between Haliotis rufescens and Haliotis discus hannai: evaluation of fertilization,larval development,growth and thermal tolerance

Two introduced abalone species are currently produced in Chile, red abalone Haliotis rufescens and Japanese abalone Haliotis discus hannai. However, red abalone accounts for 99% of total production, while the Japanese abalone has not adapted well to Chilean coastal waters. This study reports the hatching, growth and thermal tolerance performance in interspecific hybrids produced between red (R) and Japanese (J) abalone. Our results show that egg age and sperm concentration were critical factors to produce hybrids. The cross R♀ × J♂ showed a fertilization rate of 55.3 ± 3.5% using 20‐min‐old eggs and sperm concentrations of 14 × 10⁶ cells mL^?1, while the reciprocal cross (J♀ × R♂) was not successful. Further, larval development stages were similar in RR, JJ and RJ hybrid abalones. Among the experimental trials, settlement rate varied from 12.3% to 18.6% and final survival from 20.1% to 31.7%, being the RJ hybrid rates intermediate between parental species. The final shell lengths were similar between RR and RJ hybrids, but significantly higher in JJ abalones. In addition, thermal tolerance was ascertained due its pivotal role for the abalone physiology. Thus, RJ hybrids showed the highest HSP70 gene expression and offers new possibilities to expand Chilean abalone production in warm waters zones.     

Coping with climate change: Phenotypic plasticity in an imperilled freshwater fish in response to elevated water temperature

1. Climate change has emerged as an increasingly important threat to freshwater systems. To cope with rapidly changing thermal regimes, freshwater fishes must either relocate or adjust through genetic adaptation and/or phenotypic plasticity. Short-term responses to elevated water temperature have been well studied in freshwater fishes; however, far less is understood about change induced by long-term exposure. Furthermore, few studies have investigated the effects of temperature on already imperilled species, which may be more sensitive to environmental change.
2. This study investigated the effects of rearing temperature on critical thermal maximum (CT_max), agitation temperature (T_ag, temperature at which fish show behavioural signs of thermal stress) and gill size in pugnose shiner, Notropis anogenus, a threatened species in Canada. Juvenile pugnose shiner were reared for 4 months across five different ecologically relevant temperatures. CT_max and T_ag were measured under normoxia and acute exposure to hypoxia to test for oxygen sensitivity of the upper thermal limits in this species.
3. CT_max increased with elevated water temperature. T_ag also increased with rearing temperature and occurred, on average, 4.3°C above acclimation temperatures. The CT_max and T_ag were lower when fish were exposed acutely to hypoxia. Interestingly, gill size (e.g. total gill filament length) increased with rearing temperature, which may increase oxygen uptake capacity and support increased metabolic demands of warmer waters.
4. Overall, pugnose shiner show plasticity in several traits in response to long-term exposure to elevated water temperature that may facilitate persistence in warmer waters. However, acute hypoxia exposure reduced thermal tolerance, stressing the importance of evaluating interactive effects of multiple stressors.
5. Identifying source populations of pugnose shiner with greater thermal tolerance or implementing captive breeding under higher temperature regimes may improve the success of re-introduction efforts in the face of climate change, but the consequences to fitness of increased thermal tolerance should be examined.