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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 fish were continuously monitored during these swim trials, and only periods when fish were actively swimming were used in data analysis. Fish were given 1 h of recovery after the swim trial, then, they were removed from the swim tunnel and euthanized. At each swimming speed, video was recorded for 30 s from the side of the swim tunnel using a GoPro (Model HERO5; San Mateo, CA) mounted on a tri-pod. From these videos, the number of full tail oscillations in 10 s was recorded, and this value was multiplied by 6 to get the fish’s tail beat frequency (TBF) in beats min−1. Three values were averaged to calculate the mean TBF of each fish at each swimming speed.
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After surgery, the salmon were recovered in anesthetic-free water and returned to their holding tank (see below). Following all experiments, the fish were euthanized in seawater containing 0.3 g L−1 MS-222 to perform post-mortem dissections and to recover the tags/data. Post-mortem dissections were conducted to record incision length, the distance from the front of the tag to the pericardium, any signs of infection or inflammation, and to determine the fish’s sex when possible. Data were retrieved using the COM-BOX and Mercury software.
In this study, diurnal variations in fH were recorded in salmon immediately following surgery and they were maintained for the 7 and 42 day holding periods in Experiments #2 and #3, respectively (Figs. 5, 6, 7). The mean difference in fH between day-time and night-time was ~ 7 bpm and the range of values over 24 h was ~ 14 bpm (Fig. 4). This latter value is very similar to the diurnal variation in fH reported by Hvas et al. for Atlantic salmon implanted with milli-HRT tags and held at 9 °C. Conversely, Brijs et al. reported that diel variations in fH were not apparent until 3 days post-surgery in rainbow trout, and that the average daily fluctuation in fH was ~ 27 bpm. In Experiment #2, diurnal patterns in EA were present immediately following surgery and were maintained throughout the 1 week holding period at 10–11 °C (Fig. 5). In contrast, there was no evidence of significant diel variations in EA when salmon were held for 6 weeks at 8–12 °C in Experiment #3 (Fig. 6). There are two possible explanations for the discrepancy in the presence of diel patterns of activity. First, the tags were programmed to record less frequently in Experiment #3 than #2 to save memory and battery life during the 6-week holding period (i.e., every 2 h vs. 10 min or, 12 measurements per day for each fish vs. 144). This reduction in the frequency of data collection likely limited the tag’s ability to detect diel variations in EA. If the goal of future research is to study such fine scale patterns in swimming/behavior, such as diel variations of activity in fish, it is imperative that researchers optimize their sampling rate given the length of experiment they intend to perform. Second, the two trials of Experiment #3 were also conducted at colder temperatures, and the salmon were less active overall. For example, average EA was 9.5 mg (1.04 BL s−1) in Experiment #2, whereas it was only ~ 6.4 mg (0.62 BL s−1) in the longer experiments.
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Лазерное и световое лечение акне — эффективная процедура, не нарушающая целостность кожи, но при этом позволяющая глубоко проникнуть в дерму, нормализуя выработку кожного сала и уничтожая вызывающие акне и воспаления бактерии.
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