<p><b>Methodology</b><br></p>
<p>Ethical approval (REC185-19) was obtained from the Animal
Ethics Committee of the University of Pretoria. This study was a quantitative
randomised control trial during field conditions. The racing trainer selected 30 horses that
qualified for the study. The mean (±SD) age of Group A was 4 ± 0.8 years. In
Group A there were five fillies, three colts and eight geldings. In this group
one horse was eliminated due to injury. The control group had a mean (±SD) age
of 3 ± 0 years and consisted of seven geldings and eight colts with no fillies.
All participants were kept in uniform
conditions within one stable yard, received identical nutrition and training. The
researcher randomly allocated these horses into an acupuncture treatment group
(A) and a control or non-treatment group (B), with 15 horses in each group. All
individuals involved in the study, except the researcher, were blinded to which
group was the control or treatment group.
</p>
<p>The researcher performed a clinical health examination on
each horse participating in the project and the horses were habituated to the
equipment. The HRV and SP were measured pre-treatment and post-treatment, i.e.,
after the treatment group received three dry needle acupuncture treatments
spaced evenly over a three-week period. </p>
<p>The dry needle
acupuncture technique was performed by inserting acupuncture needles into
pre-selected acupuncture points (GB21, ST36, LI10, Bai Hui, Shen Shu, Shen
Peng, Shen Jiao, Yan Chi, BL40, GB27, LU1, BL54 and SI9). Needles were inserted
for 20 minutes and stimulated in a clockwise direction every five minutes. The
non-treatment group was haltered and kept in the stable for the same
period. </p>
<p><b>Heart Rate Variability</b></p>
<p>The Televet100
echocardiogram (ECG) device was used to record, digitally on a laptop 15
minutes of echocardiogram data. It was attached with gel on the skin surface by
four ECG electrodes (Trachsel et al., 2010), and secured with a surcingle. The data were recorded
pre- and post-treatment at the stables midmorning on a Sunday when the horses
were rested, one hour after feeding. </p>
<p>The data was downloaded from the recording monitor using
the Televet100 software, then transferred to the HRV Analysis Software 2.1 for
Windows or Kubios. The RR-intervals were
quantified (determining the various HRV parameters) using the Kubios software.
The frequency band widths in Kubios were set at 0.01- 0.06 Hz for low frequency
and 0.07-0.6 Hz for high frequency. A five-minute segment (within the 15
minutes recording) with the least number of artefacts (visually) was chosen.
This five-minute segment was then analysed using no correction, very low, low
correction, medium- and high-correction factors (CF) for each of the horses pre-
and post-treatment. The lowest CF that corrected the data without affecting the
variability was then chosen on an individual basis as recommended by the Kubios
website. Where the tachogram quality
was extremely poor the results were excluded from the analysis.</p>
<p>Using Kubios software,
the variability of the beat-to-beat (RR) intervals was quantified with the aid
of time domain, frequency domain and Poincaré plot analyses. The
following HRV indicators (SDNN, RMSSD, LF, HF, LF/HF, LF nu, HF, SD2) and heart
rate measures (mean RR, mean HR) were determined and statistically analysed. </p>
<p><b>Stride parameters</b></p>
<p>A GPS girth-mounted device EquinITy™ was used to measure
stride parameters and running performance in a gallop over 600m. The following
was recorded by the EquinITy™ software: maximum stride length (SL Max), average
stride length (SL Avg), maximum stride frequency (SF Max), average stride count
(SC Avg), maximum stride count (SC Max), maximum speed (Max Speed), stride count
at maximum speed (SC Max Speed), stride length at maximum speed (SL Max Speed)
and average speed (Avg Speed). Once the EquinITy™ data was downloaded from the
device at the end of the training session it was processed by the EquinITy™
Technology website per horse and work session. </p>