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To check for an adaptive component it is necessary to repeat the temperature trial on other geographically distant populations and/or sister taxa.
Upon completion of the temperature trial, fish were sacrificed using AQUI-S® solution [ 36] and dissected immediately to remove their livers.
The capacity of the strains to grow at high temperature (trial 4) was tested in duplicate in 4-day long experiments, in batch regime using 40 mL, 20 mm in diameter glass tubes.
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In order to investigate the combined effect of temperature and salinity on ULT, temperature trials were conducted under three levels of salinity which commonly occur along the coast of Texas (25, 35, and 45 ppt).
Humidity trials followed the protocol described above for temperature trials, except that temperature was maintained constant but relative humidity was maintained at one of four levels, presented in a random order (30, 55, 70, 80%).
Subsequent to completing all the temperature trials, we became concerned that variation in the relative humidity of the environmental chamber at different temperatures might be a contributing factor to variation in estimates of maximum clinging ability.
However, the respirometry trials were shorter than body temperature trials.
In this study, the production rate and specific productivity parameters were taken into account for the performance of temperature trials.
Fish were transported live to Flinders University animal rearing facility and acclimatised at a temperature of 21°C for a period of 30 days prior to the start of the temperature trials.
Aeration experiments were carried out under fully aerobic (12 L air h−1 = 0.2 vvm), fully anaerobic (12 L nitrogen h−1 = 0.2 vvm), and semiaerobic (3 L air h−1 = 0.05 vvm) conditions at the temperature value which was selected as best in temperature trials.
The majority (62.6%) of studies only measured metabolic rate at one temperature per trial, while others (19.2%) have measured RMRTa at various temperatures within a single trial period.
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