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External acceleration (EA) and variation in EA (VAR) measured in Atlantic salmon during a critical swim speed (Ucrit) test (water velocity increments of 0.2 BL s−1). Tail beat frequency (beats min−1) was determined from 30-s video clips recorded during each swimming speed. Linear mixed-effects models were fit to log transformed data and equation parameters were extracted as follows: a \( }\, = \,3.6e^ }}} ,\) b \( }\, = \,4.9e^ }}} ,\) c \( }\, = \,5.0e^ }}}\) and d \( }\, = \,11.4e^ }}} .\) Data are means ± S.E.M.; n = 6 to 8

The centi-HRT ACT and milli-HRT tags were first inserted into the tag-computer interface (COM-BOX) (this unit connected to a laptop computer), and the start time, start date and sampling intervals were set using Star-Oddi’s Mercury software. These tags were then prepared for implantation by tying two pieces of black, braided, non-absorbable and non-sterile silk suture (2–0) around the body of the tag at each end, after which these tags and the surgical equipment were cleaned thoroughly and sterilized in 70% ethanol. When milli-TD tags (which record depth and temperature; see Additional file 1: Fig. S1) were also being used, these tags were programmed using Star-Oddi’s SeaStar software.

All measurements of fH were provided with a unitless measurement known as the quality index (QI) determined by the on-board tag algorithm. This parameter represents the quality of the ECG signal, where QI0 indicates that the recording was of very good quality, QI1 and QI2 indicate decreasing quality, and QI3 indicates that no R–R interval was detected (i.e., the R peak in the PQRS complexes could not be clearly distinguished or two R peaks were not present in the ECG (for a typical ECG recording see Additional file 1: Fig. S2). ECGs were stored (which is an option when programming the DSTs) in all experiments to allow for the manual calculation of fH. In Experiment #1, manual calculations of fH from all the stored ECGs were performed. However, based on the results of Experiment #1, manual calculations of reported fH in Experiments #2 and #3 were only performed when the fish’s fH was reported to be less than 15 beats per minute (bpm), greater than 85 bpm, and/or when the QI value for fH was greater than 0. To manually calculate fH from the stored ECGs, the time between successive R wave peaks was measured (in seconds), these values were averaged, and then 60 was divided by the average to obtain the fish’s fH in bpm. Manual calculations of fH were not possible when there was only one PQRS complex or when ECG artifacts made the PQRS complex unidentifiable, and these data were not included. For Experiment #1, heart rate variability (HRV) was also calculated as the standard deviation of the time between successive R wave peaks in milliseconds (ms) .

The modified implantation method used in this study was effective for recording fH and acceleration in Atlantic salmon. In order to effectively record fH, the electrodes of fH loggers and transmitters must remain close to the pericardium throughout deployment . Therefore, we chose to suture the centi-HRT ACT tag to the body wall before closing the incision. This resulted in good quality ECG recordings during both the exhaustive exercise (Ucrit) protocol and the 1 and 6 weeks that the salmon were held in the large tanks; the average percentage of good quality ECGs (i.e., QI0) approximately 68, 86 and 88%, respectively (Table 2). It has been reported that increased activity can interfere with ECG recordings due to potentials produced by the aerobic muscles . However, this was very rare in these studies. There were only two instances when fH that could not be calculated due to noisy signals corresponded with feeding activity. The positioning of the tag was also consistent with suggestions for implanting accelerometers, e.g., aligning the tag with the major plane of movement (i.e., the lateral movement of the tail) and placing the tags close to the animal’s center of gravity . Further, suturing the tags to the body wall would have reduced the potential for variation in logger position between individuals, which can impact the interpretation of accelerometry data .

At 11:00 a.m. the following day (i.e., after 21–22 h of acclimation), a critical swim speed (Ucrit) test was performed on the fish. Specifically, the water velocity was increased to 0.6 BL s−1 (a velocity at which the fish would start swimming), and then increased by 0.2 BL s−1 every 10 min until the fish fatigued and could no longer swim. When the fish’s tail entered the back 1/6th of the tunnel, the back of the swim tunnel was tapped to encourage the fish to swim forward. In some cases (n = 5), after fish had reached their Ucrit, they were given a short (~ 5 min) rest period at low current velocity, then the water velocity was rapidly increased again to speeds above their Ucrit. From these latter trials, only accelerometry data were used (i.e., fH data were excluded).

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