Forty-seven male children aged 9 - 12 years were selected from three elementary schools in Braga, Portugal. The selection was made according to their engagement with football-oriented practice outside school context. Therefore, two groups were created, a football group (FG; n = 22) and a control group (CG; n = 25). The FG had been engaged with regular and oriented football practice in local clubs in the 9 months prior to the study, with two weekly 60-minute training sessions and sporadic weekend matches. The football training sessions consisted of a general warm-up, followed by technical exercises and several small-sided football games. The CG was not engaged with any physical activity other than complementary school-based physical education classes. Generally, school-based physical education classes consisted of a general warm-up, technical exercises of several sports, small-sided games and recreational activities lasting a total of 45 minutes. The emphasis of the physical education classes was on several different sports, according to the timing of the year and the national programme for physical education. The soccer season and the school season were coincident, and had the duration of 10 months. The parents or legal guardians provided written informed consent. The local university ethics committee approved ethical consent. All procedures were conducted according the declaration of Helsinki. The evaluation protocol included HRV measurement, anthropometry, physical activity and physical fitness tests. Testing took place in two interspersed days for each school. One session was dedicated to HRV measurements; the other session was devoted to anthropometry and physical fitness assessment. HRV was measured using Polar Team 2 TM (Polar Electro OY, Kempele, Finland) heart monitors set to record R-R intervals. HRV recording took place at 9:00 in a quiet and silent room, with low light, comfortable temperature and in a fast. The children were in the supine position; no visual contact was granted. The recording lasted 15 minutes, preceded by a 5-minutes explanation. Metronomic breathing was discarded because children have difficulty pacing their breathing with a pre-determined cadence, and therefore potential cardiac alterations might occur (
25,
26). Raw data was converted to ASCII format in the form of R-R intervals in milliseconds, and exported to Kubios HRV TM version 2.0 (Department of Physics, University of Kuopio, Kuopio, Finland). The last 5 minutes of the test were used to analyse the R-R intervals using time and frequency-domain techniques. Custom artifact correction was used, and trend components were removed according to the smoothness priors method on Kubios software. Non-parametric methods, namely FFT, were used to obtain measures of HRV frequency-domain. Information regarding total power and three-frequency bands was obtained by default: very low frequency (VLF: 0 - 0.04 Hz), low frequency (LF: 0.04 - 0.15 Hz) and high frequency (HF: 0.15 - 0.4 Hz). VLF was discarded from the analysis due to its uncertain physiological meaning and interpretation (
7). HF reflects vagal activity (
7); no agreement exists regarding LF, yet it seems to reflect mainly sympathetic modulation, especially when normalized (
7). LF and HF normalized units (n.u.) were also calculated. LF:HF ratio was calculated since it might express sympathovagal balance (
7). Height (cm) (stadiometer model 708, Seca, Hamburg, Germany), weight (kg) and percent body fat (Tanita Inner Scan, BC-532, Tanita, Amsterdam, Netherlands), and physical activity questionnaires were assessed in the classroom. Lean mass (kg) was obtained by subtracting the fat mass to the weight of each individual. The short form of the international physical activity questionnaire (IPAQ) was applied to provide information regarding the time spent on moderate-to-vigorous physical activities (MVPA). The validity and reliability of this instrument was tested in several countries, including Portugal (
27). IPAQ has been often used to assess children and early adolescents' physical activity (
28-
31). The questionnaire was applied during an interview led by the physical education teacher of each student. Information regarding the daily time spent on MVPA was obtained by summing the average duration per day spent in each intensity level. Fitness tests were administered on an indoor multi-sports ground. Prior to testing, the participants performed a 12-minute warm-up and familiarization trials with each test. Speed was evaluated with a 15-m sprint test. Elapsed times were obtained with photoelectric cells (Speed Trap II, Brower Timing Systems, Utah, USA) positioned at the starting line, and at 5 and 15 m. Participants sprinted from a standing position 30 cm behind the starting line. The fastest of 2 trials was considered. Jumping height was evaluated by countermovement jump (CMJ) on a special mat (Digitime 1000, Digitest, Jyvaskyla, Finland) maintaining hands on hips. The best of two trials was retained. The Yo-Yo intermittent endurance test - level 1 (Yo-Yo IE1) was used to evaluate aerobic capacity (
32). Research has proven that Yo-Yo IE1 can be used as an indicator of aerobic fitness for children of this age range (
33). The test consists of repeated 2 × 20-m shuttle runs between a start and finish line, interspersed by 5-s rest period between runs. Progressively increased speeds are controlled by audio bleeps from a CD-recorder. The aim of the test is to perform as many shuttles as possible. When the child failed twice to reach the finish line in time, the distance covered was recorded and used as the test result. One trial was given. Data was tested for normality using the Shapiro-Wilk test. Descriptive statistics consisted of mean ± standard deviation (SD); the non-normally distributed variables are presented as medians and interquartile (IQ) ranges. Differences between groups were obtained using independent-samples t-test. A Mann-Whitney test was performed to obtain differences between groups for non-normally distributed variables (LF, HF, total power). Standardised differences in means (effect sizes, d) were computed for comparisons. Effect sizes were classified according to Hopkins (
34) as trivial (d < 0.2), small (0.2 < d < 0.6) moderate (0.6 < d < 1.2), large (1.2 < d < 2.0), very large (2.0 < d < 4.0), nearly perfect (d > 4.0), and perfect (d = infinite). Statistical significance was set at P < 0.05.