Oxygen uptake during high-intensity running: response following a single bout of interval training

Elevated oxygen uptake (VO2) during moderate-intensity running following a bout of interval running training has been studied previously. To further investigate this phenomenon, the VO2 response to high-intensity exercise was examined following a bout of interval running. Well-trained endurance runners were split into an experimental group [maximum oxygen uptake, VO2max 4.73 (0.39)l x min(-1)] and a reliability group [VO2max 4.77 (0.26)l x min(-1)]. The experimental group completed a training session (4 x 800 m at 1 km x h(-1) below speed at VO2max, with 3 min rest between each 800-m interval). Five minutes prior to, and 1 h following the training session, subjects completed 6 min 30 s of constant speed, high-intensity running designed to elicit 40% delta (where delta is the difference between VO2 at ventilatory threshold and VO2max; tests 1 and 2, respectively). The slow component of VO2 kinetics was quantified as the difference between the VO2 at 6 min and the VO2 at 3 min of exercise, i.e. deltaVO2(6-3). The deltaVO2(-3) was the same in two identical conditions in the reliability group [mean (SD): 0.30 (0.10)l x min(-1) vs 0.32 (0.13)l x min(-1)]. In the experimental group, the magnitude of the slow component of VO2 kinetics was increased in test 2 compared with test 1 by 24.9% [0.27 (0.14)l x min(-1) vs 0.34 (0.08)l x min(-1), P < 0.05]. The increase in deltaVO2(6-3) in the experimental group was observed in the absence of any significant change in body mass, core temperature or blood lactate concentration, either at the start or end of tests 1 or 2. It is concluded that similar mechanisms may be responsible for the slow component of VO2 kinetics and for the fatigue following the training session. It has been suggested previously that this mechanism may be linked primarily to changes within the active limb, with the recruitment of alternative and/or additional less efficient fibres.     

Oxygen uptake during moderate intensity running: response following a single bout of interval training

Eight male endurance runners [mean ± (SD): age 25 (6) years; height 1.79 (0.06) m; body mass 70.5 (6.0) kg; % body fat 12.5 (3.2); maximal oxygen consumption (V˙O_2max 62.9 (1.7) ml · kg⁻¹ · min⁻¹] performed an interval training session, preceded immediately by test 1, followed after 1 h by test 2, and after 72 h by test 3. The training session was six 800-m intervals at 1 km · h⁻¹ below the velocity achieved at V˙O_2max with 3 min of recovery between each interval. Tests 1, 2 and 3 were identical, and included collection of expired gas, measurement of ventilatory frequency (f _v ), heart rate (f _c), rate of perceived exertion (RPE), and blood lactate concentration ([La⁻]_B) during the final 5 min of 15 min of running at 50% of the velocity achieved at V˙O_2max (50% −V˙O_2max).␣Oxygen uptake (V˙O₂), ventilation (V˙ _E ), and respiratory exchange ratio (R) were subsequently determined from duplicate expired gas collections. Body mass and plasma volume changes were measured preceding and immediately following the training session, and before tests 1–3. Subjects ingested water immediately following the training session, the volume of which was determined from the loss of body mass during the session. Repeated measures analysis of variance with multiple comparison (Tukey) was used to test differences between results. No significant differences in body mass or plasma volume existed between the three test stages, indicating that the differences recorded for the measured parameters could not be attributed to changes in body mass or plasma volume between tests, and that rehydration after the interval training session was successful. A significant (P < 0.05) increase was found from test 1 to test 2 [mean (SD)] for V˙O₂ [2.128 (0.147) to 2.200 (0.140) 1 · min⁻¹], f _c [125 (17) to 132 (16) beats · min⁻¹], and RPE [9 (2) to 11 (2)]. A significant (P < 0.05) decrease was found for submaximal R [0.89 (0.03) to 0.85 (0.04)]. These results suggest that alterations in V˙O₂ during moderate-intensity, constant-velocity running do occur following heavy-intensity endurance running training, and that this is due to factors in addition to changed substrate metabolism towards greater fat utilisation, which could explain only 31% of the increase in V˙O₂. Accepted: 8 December 1997     

A three component model of human bioenergetics

The model described in this article is a generalised three component hydraulic model, proposed to represent net whole body bioenergetic processes during human exercise and recovery. During exercise, fluid flows from the three interconnected vessels in the system represent the breakdown of high energy phosphates (phosphagens), oxygen consumption and lactic acid production. During recovery, replenishment of the fluids represents the repayment of oxygen debt. The model is quantified and solved mathematically, and the solution compared with observed experimental data. Since currently known physiological facts are consistent with four configurations of this model, further experimentation is necessary.     

Effects of running training on in vitro brown adipose tissue thermogenesis in rats

Brown adipose tissue (BAT) is a major site of nonshivering thermogenesis (NST) during cold acclimation for most mammals. Repetitive nonthermal stress such as immobilization has been shown to enhance the capacity of NST as cold acclimation. In the present study, the effects of running training, another type of nonthermal stress, were investigated on in vitro thermogenesis and the cellularity of interscapular BAT in rats. The rats were subjected to treadmill running for 30 min daily at 30 m/min under 8° inclination for 4–5 weeks. In vitro thermogenesis was then measured in minced tissue blocks incubated in a Krebs-Ringer phosphate buffer containing glucose and albumin at 37° C, using a Clark type oxygen electrode. The trained rats showed less body weight gain during the experiment. The weights of BAT and epididymal white adipose tissue were smaller in the trained rats. Noradrenaline- and glucagon-stimulated oxygen consumption were also significantly smaller in the trained rats. The tissue DNA level was greater in the trained rats, but the DNA content per tissue pad did not significantly differ. The results indicate that running training reduces BAT thermogenesis, possibly as an adaptation to conserve energy substrates for physical work.     

Assessment of running velocity at maximal oxygen uptake

The purpose of the study was to compare the cardiovascular, respiratory and metabolic responses to exercise of highly endurance trained subjects after 3 different nights i.e. a baseline night, a partial sleep deprivation of 3 h in the middle of the night and a 0.25-mg triazolam-induced sleep. Sleep-waking chronobiology and endurance performance capacity were taken into account in the choice of the subjects. Seven subjects exercised on a cycle ergometer for a 10-min warm-up, then for 20 min at a steady exercise intensity (equal to the intensity corresponding to 75% of the predetermined maximal oxygen consumption) followed by an increased intensity until exhaustion. The night with 3 h sleep loss was accompanied by a greater number of periods of wakefulness (P less than 0.01) and fewer periods of stage 2 sleep (P less than 0.05) compared with the results recorded during the baseline night. Triazolam-induced sleep led to an increase in stage 2 sleep (P less than 0.05), a decrease in wakefulness (P less than 0.05) and in stage 3 sleep (P less than 0.05). After partial sleep deprivation, there were statistically significant increases in heart rate (P less than 0.05) and ventilation (P less than 0.05) at submaximal exercise compared with results obtained after the baseline night. Both variables were also significantly enhanced at maximal exercise, while the peak oxygen consumption (VO2) dropped (P less than 0.05) even though the maximal sustained exercise intensity was not different.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of exercise training on brain opioid peptides and serum LH in female rats

In order to investigate the effects of long-term exercise training on brain endorphin systems, and the latter's possible effects on the hypothalamic-pituitary-gonadal axis, female Wistar rats were subjected to daily treadmill running. A sedentary control group was also employed. After 8 weeks of training, and just prior to sacrifice, one-half of each group received a final fatiguing bout of exercise. Thus the final four groups consisted of a trained-fatigued (TF), trained-nonfatigued (TN), control-fatigued (CF), and control-nonfatigued (CN) group. Regional brain levels of beta-endorphin (beta E), methionine enkephalin and leucine enkephalin (LE) were assayed with independent RIAs from the nucleus accumbens, cortex, caudate-putamen, septum, amygdala, anterior and posterior hypothalamus, substantia nigra and ventral tegmentum. Diestrus serum levels of luteinizing hormone (LH), follicle stimulating hormone and prolactin (PRL) were also determined. Fatiguing resulted in a decrease in serum LH levels as well as an increase in beta E content in the nucleus accumbens, and LE content in the ventral tegmentum. Finally, TF animals exhibited less LE in the amygdala than the TN rats. Taken together, these changes in brain endorphins may indicate an acute, fatigue-running modulation of the hypothalamic-pituitary-gonadal axis.     

Heritability of running economy: a study made on twin brothers

Running economy (RE), defined as the steady-state of oxygen uptake (V˙O₂) for a given running velocity, is a factor of sports performance the genetic component of which has seldom been reported to date. We studied this component using a heritability index (HI) in a group of 32 male twins, 8 monozygotic (MZ) and 8 dizygotic (DZ) pairs, all sportsmen with similar perinatal and environmental backgrounds. Zygocity was determined by the identity of erythrocytic antigenic, protein and enzymatic polymorphism, and human leucocyte antigen serologic types between co-twins. The subjects exercised twice on a treadmill, once until exhaustion and again at submaximal intensities. Pulmonary gas exchange was measured continuously using an automatic analyser system during both tests. Blood samples were obtained during the recovery period to determine lactate concentrations. No significant differences were observed between MZ and DZ, in respect of RE at any speed or in maximal V˙O₂ relative to body mass. Nevertheless, significant HI (P < 0.05) was found in maximal lactate concentrations (HI = 0.75) and in respiratory equivalent for oxygen at two speeds, 7 km · h⁻¹ (HI = 0.71) and 8 km · h⁻¹ (HI = 0.79), differences which probably suggest that there are differences in RE. In conclusion, we did not detect a genetic component in RE or in maximal oxygen uptake, but a genetic component for markers of anaerobic metabolism was present. Accepted: 5 November 1997     

Oxygen uptake during running as related to body mass in circumpubertal boys: a longitudinal study

To investigate the effect of endurance training on physiological characteristics during circumpubertal growth, eight young runners (mean starting age 12 years) were studied every 6 months for 8 years. Four other boys served as untrained controls. Oxygen uptake (VO2) and blood lactate concentrations were measured during submaximal and maximal treadmill running. The data were aligned with each individual's age of peak height velocity. The maximal oxygen uptake (VO2max; ml.kg-1.min-1) decreased with growth in the untrained group but remained almost constant in the training group. The oxygen cost of running at 15 km.h-1 (VO2 15, ml.kg-1.min-1) was persistently lower in the trained group but decreased similarly with age in both groups. The development of VO2max and VO2 15 (l.min-1) was related to each individual's increase in body mass so that power functions were obtained. The mean body mass scaling factor was 0.78 (SEM 0.07) and 1.01 (SEM 0.04) for VO2max and 0.75 (SEM 0.09) and 0.75 (SEM 0.02) for VO2 15 in the untrained and trained groups, respectively. Therefore, expressed as ml.kg-0.75.min-1, VO2 15 was unchanged in both groups and VO2max increased only in the trained group. The running velocity corresponding to 4 mmol.l-1 of blood lactate (nu la4) increased only in the trained group. Blood lactate concentration at exhaustion remained constant in both groups over the years studied. In conclusion, recent and the present findings would suggest that changes in the oxygen cost of running and VO2max (ml.kg-1.min-1) during growth may mainly be due to an overestimation of the body mass dependency of VO2 during running.(ABSTRACT TRUNCATED AT 250 WORDS)     

Assessment of middle-distance running performance in sub-elite young runners using energy cost of running

The purpose of this study was to assess the validity of v _amax as an indicator of middle-distance running performance in sub-elite young runners, _amax being defined as the quotient maximal oxygen uptake (V˙O _2max) divided by the net energy cost of running (C _r) on a treadmill at a submaximal running velocity (280 m · min⁻¹). The V˙O _2max, ventilatory threshold, _amax, and C _r were assessed in 39 young male sub-elite runners having only small variations in performance level. The relationship between each variable and running performance (at 1500 m, 3000 m, and 5000 m) was evaluated. A trend toward a negative correlation existed between C _r and performance although this was not significant. The V˙O _2max and _amax were significantly related to performance. The _amax accounted for around 50% of the variability in performance whereas other physiological variables selected in this study were responsible, at best, for approximately 39%. The results presented in this study suggested that _amax was a useful indicator of middle-distance running performance in sub-elite young runners with similar performance levels as well as in top elite athletes. Accepted: 19 August 1997     

Effect of stride frequency on thermoregulatory responses during endurance running in distance runners

Changing stride frequency may influence oxygen uptake and heart rate during running as a function of running economy and central command. This study investigated the influence of stride frequency manipulation on thermoregulatory responses during endurance running. Seven healthy endurance runners ran on a treadmill at a velocity of 15 km/h for 60 min in a controlled environmental chamber (ambient temperature 27 °C and relative humidity 50%), and stride frequency was manipulated. Stride frequency was intermittently manipulated by increasing and decreasing frequency by 10% from the pre-determined preferred frequency. These periods of increase or decrease were separated by free frequency running in the order of free stride frequency, stride frequency manipulation (increase or decrease), free stride frequency, and stride frequency manipulation (increase or decrease) for 15 min each. The increased and decreased stride frequencies were 110% and 91% of the free running frequency, respectively (196±6, 162±5, and 178±5 steps/min, respectively, P<0.01). Compared to the control, stride frequency manipulation did not affect rectal temperature, heart rate, or the rate of perceived exhaustion during running. Whole-body sweat loss increased significantly when stride frequency was manipulated (1.48±0.11 and 1.57±0.11 kg for control and manipulated stride frequencies, respectively, P<0.05), but stride frequency had a small effect on sweat loss overall (Cohen's d=0.31). A higher mean skin temperature was also observed under mixed frequency conditions compared to that in the control (P<0.05). While the precise mechanisms underlying these changes remain unknown (e.g. running economy or central command), our results suggest that manipulation of stride frequency does not have a large effect on sweat loss or other physiological variables, but does increase mean skin temperature during endurance running.     

Effect of running training on uncoupling protein mRNA expression in rat brown adipose tissue

The effect was investigated of endurance training on the expression of uncoupling protein (UCP) mRNA in brown adipose tissue (BAT) of rats. The exercised rats were trained on a rodent treadmill for 5 days per week and a total of 9 weeks. After the training programme, a marked decrease in BAT mass was found in terms of weight or weight per unit body weight; there was a corresponding decrease in DNA content and a downward trend in RNA and glycogen levels. The UCP mRNA was present at a markedly decreased level in BAT of trained animals. In consideration of the reduced levels of mRNAs for hormone-sensitive lipase and acylCoA synthetase, the brown adipose tissue investigated appeared to be in a relatively atrophied and thermogenically quiescent state.     

The longitudinal development of running economy in males and females aged between 13 and 27 years: The Amsterdam Growth and Health Study

The purpose of this study was to describe the longitudinal development of running economy [defined as the oxygen uptake (V˙O₂) at a submaximal running speed] in males and females from teenage to young adult age using data from the Amsterdam Growth and Health Study. Submaximal V˙O₂ (in ml · kg⁻¹· min⁻¹) was measured in 84 males and 98 females while they ran on a treadmill at a constant speed of 8 km · h⁻¹ for 6 min at three different treadmill slopes (0%, 2.5% and 5%). This test was carried out six times, on the same subjects at the ages of 13, 14, 15, 16, 21, and 27 years. The longitudinal development of running economy in males and females was analysed using a two-way analysis of variance for repeated measurements. At all three slopes, a significant decrease in V˙O₂ with increasing age was found for both males and females, implying a significant increase in running economy for both sexes. Males showed significantly higher V˙O₂ values than females at all ages measured and for all three slopes, suggesting that females have a significantly higher running economy than males. In order to make a better comparison of the V˙O₂ of individuals of different sizes, allometric models were used; power function ratios were constructed in which body mass was expressed to an exponential power. Following this analysis the difference in submaximal V˙O₂ and running economy between males and females appeared even larger. Accepted: 20 February 1997     

Better economy in field running than on the treadmill: evidence from high-level distance runners

Given the ongoing interest in ways to improve the specificity of testing elite athletes in their natural environment, portable metabolic systems provide an opportunity to assess metabolic demand of exercise in sport-specific settings. Running economy (RE) and maximal oxygen uptake (V.O₂max) were compared between track and treadmill (1% inclination) conditions in competitive level European distance runners who were fully habituated to treadmill running (n = 13). All runners performed an exercise test on running track and on treadmill. While V.O₂max was similar on the track and on the treadmill (68.5 ± 5.3 vs. 71.4 ± 6.4 ml·kg⁻¹·min⁻¹, p = 0.105, respectively), superior RE was found on the track compared to the treadmill (215.4 ± 12.4 vs. 236.8 ± 18.0 O₂ ml·kg⁻¹·km⁻¹, p < 0.001). RE on the track was strongly correlated with RE on the treadmill (r = 0.719, p = 0.006). The present findings indicate that high-level distance runners have significantly better RE but not V.O₂max on the track compared to treadmill. This difference may be due to biomechanical adjustments. As RE is strongly correlated between the two conditions, it would be reasonable to assume that interventions affecting RE on the treadmill will also affect RE on the track.     

Detection of the change point in oxygen uptake during an incremental exercise test using recursive residuals: relationship to the plasma lactate accumulation and blood acid base balance

The purpose of this study was to develop a method to determine the power output at which oxygen uptake (V˙O₂) during an incremental exercise test begins to rise non-linearly. A group of 26 healthy non-smoking men [mean age 22.1 (SD 1.4) years, body mass 73.6 (SD 7.4) kg, height 179.4 (SD 7.5) cm, maximal oxygen uptake (V˙O_2max) 3.726 (SD 0.363) l · min⁻¹], experienced in laboratory tests, were the subjects in this study. They performed an incremental exercise test on a cycle ergometer at a pedalling rate of 70 rev · min⁻¹. The test started at a power output of 30 W, followed by increases amounting to 30 W every 3 min. At 5 min prior to the first exercise intensity, at the end of each stage of exercise protocol, blood samples (1 ml each) were taken from an antecubital vein. The samples were analysed for plasma lactate concentration [La]_pl, partial pressure of O₂ and CO₂ and hydrogen ion concentration [H⁺]_b. The lactate threshold (LT) in this study was defined as the highest power output above which [La⁻]_pl showed a sustained increase of more than 0.5 mmol · l⁻¹ · step⁻¹. The V˙O₂ was measured breath-by-breath. In the analysis of the change point (CP) of V˙O₂ during the incremental exercise test, a two-phase model was assumed for the 3rd-min-data of each step of the test: X _i =at _i +b+ɛ _i for i=1,2,…,T, and E(X _i )>at _i +b for i =T+1,…,n, where X ₁, … , X _n are independent and ɛ _i ∼N(0,σ²). In the first phase, a linear relationship between V˙O₂ and power output was assumed, whereas in the second phase an additional increase in V˙O₂ above the values expected from the linear model was allowed. The power output at which the first phase ended was called the change point in oxygen uptake (CP-V˙O₂). The identification of the model consisted of two steps: testing for the existence of CP and estimating its location. Both procedures were based on suitably normalised recursive residuals. We showed that in 25 out of 26 subjects it was possible to determine the CP-O₂ as described in our model. The power output at CP-V˙O₂ amounted to 136.8 (SD 31.3) W. It was only 11 W – non significantly – higher than the power output corresponding to LT. The V˙O₂ at CP-V˙O₂ amounted to 1.828 (SD 0.356) l · min⁻¹ was [48.9 (SD 7.9)% V˙O_{2 max} ]. The [La⁻]_pl at CP-V˙O₂, amounting to 2.57 (SD 0.69) mmol · l⁻¹ was significantly elevated (P<0.01) above the resting level [1.85 (SD 0.46) mmol · l⁻¹], however the [H⁺]_b at CP-V˙O₂ amounting to 45.1 (SD 3.0) nmol · l⁻¹, was not significantly different from the values at rest which amounted to 44.14 (SD 2.79) nmol · l⁻¹. An increase of power output of 30 W above CP-V˙O₂ was accompanied by a significant increase in [H⁺]_b above the resting level (P=0.03). Accepted: 25 March 1998     

Effect of increasing running velocity on electroencephalogram in a field test

This study was designed to measure the electroencephalogram (EEG) after exercise with increasing intensity. In a field test with increments in running velocity a 2-min EEG was recorded, together with blood lactate concentration and heart rate, after each stage. An individual protocol was used, with up to six stages of running to ensure comparability of exercise intensity among the subjects, in each of 19 athletes (17 men, 2 women) experienced in leisure-time running. The exercise consisted initially of three running stages of aerobic exercise intensity without blood lactate accumulation followed by stages with an increase of lactate concentration. The protocol of the field test led to a progressive increase in cortical activity directly after the stages without blood lactate accumulation mainly in the δ frequency band, followed by θ and α-1 frequency band, and less pronounced in the α-2 and in the β frequency bands. After the stages with an onset and further increase of blood lactate accumulation significant decreases in the β-2, β-1 and α-1 frequency bands occurred predominantly in temporal (T3, T4, T5, and T6) and occipital (O1, and O2) electrode positions, indicating a stage-by-stage decrease of activity. This decrease may be explained by feed-back from working muscle, via afferents to the cortex from intero- and proprio-receptors and affective processes. This could suggest that through a higher running intensity indicated by an onset of blood lactate accumulation metabolic and mechanical changes led to alterations within the afferent systems influencing the level of cortical activity. Accepted: 9 February 1998     

Effect of extra running on running-based taste aversion in rats

Voluntary running in an activity wheel endowed rats with aversion to a taste solution consumed before the running. This running-based taste aversion was attenuated by extra running opportunities interspersed among the taste–running pairings, but the attenuating effect was reduced by signaling the extra running by another taste cue. These results correspond to the so-called degraded contingency effect and cover-cue effect in the traditional preparations of Pavlovian conditioning.     

Effects of specific versus cross-training on running performance

The cross-training (XT) hypothesis suggests that despite the principle of specificity of training, athletes may improve performance in one mode of exercise by training using another mode. To test this hypothesis we studied 30 well-trained individuals (10 men, 20 women) in a randomized longitudinal trail. Subjects were evaluated before and after 8 weeks of enhanced training (+10%/week), accomplished by adding either running (R) or swimming (XT) to baseline running, versus continued baseline running (C). Both R ( – 26.4s) and XT (– 13.2s) improved time trial (3.2 km) performance, whereas C did not (– 5.4s). There were no significant changes during treadmill running in maximum oxygen uptake (O_2peak; – 0.2, – 6.0, and + 2.7%), steady state submaximal O₂ at 2.68 m · s^–1 ( – 1.2, – 3.3 and + 0.2 ml · kg^–1 · min^–1), velocity at O_2peak (+0.05, +0.25 and +0.09 m · s^–1) or accumulated O₂ deficit (+ 11.2, – 6.1 and + 9.4%) in the R, XT or C groups, respectively. There was a significant increase in velocity associated with a blood lactate concentration of 4 mmol · l^–1 in R but not in XT or C ( + 0.32, + 0.07 and + 0.08 m · s^–1). There were significant changes in arm crank O_2peak ( + 5%) and arm crank O₂ at 4 mmol · l^–1 ( + 6.4%) in XT. There was no significant changes in arm crank O_2peak ( + 1.3 and – 7.7%) or arm crank O₂ at 4 mmol · l^–1 ( + 0.8 and + 0.4%) in R or C, respectively. The data suggest that muscularly non-similar XT may contribute to improved running performance but not to the same degree as increased specific tranining.     

Effect of step width manipulation on tibial stress during running

Narrow step width has been linked to variables associated with tibial stress fracture. The purpose of this study was to evaluate the effect of step width on bone stresses using a standardized model of the tibia. 15 runners ran at their preferred 5 k running velocity in three running conditions, preferred step width (PSW) and PSW±5% of leg length. 10 successful trials of force and 3-D motion data were collected. A combination of inverse dynamics, musculoskeletal modeling and beam theory was used to estimate stresses applied to the tibia using subject-specific anthropometrics and motion data. The tibia was modeled as a hollow ellipse. Multivariate analysis revealed that tibial stresses at the distal 1/3 of the tibia differed with step width manipulation (p=0.002). Compression on the posterior and medial aspect of the tibia was inversely related to step width such that as step width increased, compression on the surface of tibia decreased (linear trend p=0.036 and 0.003). Similarly, tension on the anterior surface of the tibia decreased as step width increased (linear trend p=0.029). Widening step width linearly reduced shear stress at all 4 sites (p<0.001 for all). The data from this study suggests that stresses experienced by the tibia during running were influenced by step width when using a standardized model of the tibia. Wider step widths were generally associated with reduced loading of the tibia and may benefit runners at risk of or experiencing stress injury at the tibia, especially if they present with a crossover running style.     

Effect of high boron levels on growth and some metabolic activities of the halotolerantdunaliella tertiolecta

Growth of Dunaliella tertiolecta was retarded when the alga was exposed to high boron (B) concentrations between 50 and 200 g m-³. Photosynthetic oxygen evolution as well as respiratory oxygen uptake were significantly suppressed with the rise of B concentration. Similarly the contents of chlorophyll, glycerol, lipid and proteins were lowered, while that of saccharides was raised.