BIOM2012 report Introduction:

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Previous studies have described the pattern of the ventilatory response during steady-state exercise as a linear relationship with increasing work rate (Burton, et al. 2004). The results of Wasserman (1978) study found that exercise above 60% of the subjects maximal work capacity resulted in a curvilinear reflection upward during minute ventilation suggesting an increased metabolic demand for higher intensity exercises. It is generally accepted that the rise in ventilation following exercise is attributed to increased metabolic rates of oxygen uptake and carbon dioxide output (Wasserman, 1978). Thus it follows that carbon dioxide tension in the systemic circulation and in the brain are major determinants of breathing during higher metabolic demand (Blain, 2010). This idea is reinforced by Beaver & Wasserman (1970) study which found an uniform relationship between minute ventilation and carbon dioxide output during higher work capacities. Wasserman (1978) argues that acidosis, which is the induced production of lactic acid only occurs under heavy work intensities and is compensated by hyperventilation causing the arterial carbon dioxide tension to become subnormal. Thus it is hypothesised that in humans, participation in high intensity cardiovascular exercise will result in a greater increase in alveolar ventilation rate (AV) compared to low intensity cardiovascular exercise. Geppert and Zuntz (1888) first suggested that the regulatory mechanism which controls this ventilatory response under exercise was transmitted via a neural or humoral pathway. However the results of Kao (1956) cross-circulation perfusion experiment concluded that humoral factors were not involved and only the neural pathway exists, consisting of motor centre activity and afferent signals from chemoreceptors. The peripheral chemoreceptors, primarily the carotid bodies detect the changes in PaO2 and PaCO2 signalling the respiratory centre in the brain to increase ventilation (Blain, 2010). Wasserman (1978) also found that the increased ventilation rate during metabolic acidosis was significantly reduced in the absence of carotid bodies. However Whipp & Ward (1998) study argues that the change in PaO2 and PaCO2 does not fluctuate sufficiently to stimulate the ventilatory response in exercise, suggesting that the real determinant is from the sensitivity of peripheral chemoreceptors to oscillations in PaO2 and PaCO2. In a similar process, central chemoreceptors in the medulla oblongata detect the changes in acidity caused by carbon dioxide liberation during exercise, stimulating the respiratory centre to increase ventilation (Wasserman, 1978). Despite intensive research for over a century, there has been no general consensus into the mechanisms of respiratory control during exercise, as competing theories exist on whether peripheral and central chemoreceptors have additive or independent effects to the overall ventilatory response (Blain, 2010). The results of Beaver & Wasserman (1970) study found considerable oscillations in respiratory rate and tidal volume, the two primary determinants of alveolar ventilation rate. Contradictory results from Phillipson et al. (1970) found that this vagal reflex did not alter the ventilatory response to exercise but rather altered the breathing pattern, as the increase in

ventilation was primarily the result of an increase in tidal volume rather than in respiratory rate (Phillipson et al. 1970) .Nonetheless, Wasserman (1978) study concluded that rises in respiratory rate and tidal volume did occur during heavy working capacities when metabolic acidosis was induced. Materials & Methods: Subjects Nineteen subjects (11 male subjects and 8 female subjects) volunteered to participate in the study. The subjects reported good health with no previous cardiovascular or respiratory illness. Subjects were required to give written and informed consent for the study. To limit external variables, subjects were instructed to avoid exercise and maintain regular dietary and hydration habits. Experimental Set-up A filter and clean mouthpiece was attached to the end of the spirometer for each subject. A nose clip was used to ensure all air passed through the mouthpiece, filter and flow head. The two parameters recorded were respiratory rate and tidal volume with the aid of the program LabChart (AdInstruments). Experimental Protocol Subjects were randomly allocated to either a high intensity exercise or a low intensity exercise condition. Intensity was based on preset cycle ergometer resistance levels with low intensity = 0 and high intensity = 3. The subjects heart rate was also monitored at least once every minute during exercise to ensure it did not exceed 180 bpm. Prior to exercise, subjects were required to measure their baseline tidal volume and respiratory rate for one minute using a spirometer. Subjects cycled on an cycle ergometer for 5 minutes at their individual allocated intensity maintaining a speed of 25km/hr for resistance = 0 and 30km/hr for resistance = 3. After exercise the subjects tidal volume and respiratory rate were measured at times; 10minutes, 15minutes and 20minutes post test. Subjects were then required to repeat the entire procedure with the alternate intensity condition the following week, controlling for between-subject variation. Data analysis Alveolar ventilation rate (AV) was calculated from data collected for tidal volume (TV) and respiratory rate (RR) with the equation AV = (TV DS) x RR, assuming dead space (DS) of 150mL. After the data was collected, a two-way ANOVA analysis of alveolar ventilation rate, tidal volume and respiratory rate was conducted using the program GraphPad Prism 5 to detect statistical significance (P<0.05). The Bonferroni post-hoc test was then used to find statistical significance for multiple comparisons of time for each parameter.

Results: There was an overall initial rising trend in mean AV (Fig 1.), mean TV (Fig 2.) and mean RR (Fig 3.) from t = 0-10min, followed by an overall decreasing trend from t=10-20min. A twoway ANOVA analysis revealed no statistical significance (P>0.05) in AV (Fig 1.) and TV (Fig 2.) observed for baseline recordings (t = 0min) and endpoint recordings (t =15min, t=20min) between high intensity and low intensity groups. However the Bonferroni post-hoc test revealed statistical significance (P<0.05) in AV (Fig 1.) and TV (Fig 2.) observed immediately post-exercise (t=10min), between high intensity and low intensity groups. No statistical significance (P>0.05) in RR (Fig 3.) was observed for baseline recordings (t = 0min), immediately post-exercise (t=10min) and endpoint recordings (t =15min, t=20min) between high intensity and low intensity groups.

Figure 1. Mean SEM Alveolar Ventilation (mL/min) recorded for low intensity exercise and high intensity exercise over the duration of 20 minutes (n=19). Measurements were taken at 0 min for baseline recording, and post-exercise at 10, 15 and 20 min. Different superscript letters denote statistically significantly different values for conditions (P<0.05). Two-way error bars are plotted using standard error of mean.

Figure 2. Mean SEM Tidal Volume (mL) recorded for low intensity exercise and high intensity exercise over the duration of 20 minutes (n=19). Measurements were taken at 0 min for baseline recording, and post-exercise at 10, 15 and 20 min. Different superscript letters denote statistically significantly different values for conditions (P<0.05). Two-way error bars are plotted using standard error of mean.

Figure 3. Mean SEM Respiration Rate (breaths/min) recorded for low intensity exercise and high intensity exercise over the duration of 20 minutes (n=19). Measurements were taken at 0 min for baseline recording, and post-exercise at 10, 15 and 20 min. Different superscript letters denote statistically significantly different values for conditions (P<0.05). Two-way error bars are plotted using standard error of mean.

Discussion: The results obtained from the study support our hypothesis that high-intensity exercise will result in a significantly greater increase in AV immediately post-exercise than low-intensity exercise as evidenced in the increasing trend in AV with a significant difference observed between high-intensity and low-intensity groups (Fig. 1). Upon further analysis it is found that the increasing trend is attributed to the significantly greater increase in TV immediately post-exercise in high intensity exercise than low intensity exercise, as TV is one of the primary determinants of AV (Fig. 2). It is also observed that there was no contribution from RR rate towards the greater increase in AV between high-intensity and low-intensity groups, as there was no significant difference observed immediately post-exercise between highintensity and low-intensity groups for RR (Fig. 3). This suggests that at high-intensity exercise the underlying determinant for the significant increase in AV is from the significant increase in TV. Whipp & Ward (1998) study supports the idea of exercise hyperpnoea, arguing that increased cellular oxygen requirement and carbon dioxide production during exercise promotes increased TV, allowing venous blood to be oxygenated and the maintenance of arterial carbon dioxide tension and hydrogen-ion homeostasis. One proposed mechanism for exercise hyperpnoea in relation to respiratory control involves proprioceptors or muscle spindles inside exercising muscles which argues that increased AV observed at the start of exercise is governed by a set of impulses carried by nerves from the exercising muscle (Wasserman, 1978). Results from Beaver & Wasserman (1970) study demonstrated that abrupt changes in TV and RR existed at the start and cessation of exercise with TV and RR changing reciprocally rather than complementarily. Conversely other studies have shown during steady-state exercise TV and RR increases evenly, however under heavy-intensity exercise TV increases more than RR till the induction of metabolic acidosis where the increases in RR predominate (Wasserman, 1978). This pattern contradicts with our results, which indicates a steady increasing trend in both TV and RR at the same time interval at t = 0-10 for both high and low intensity groups (Fig. 2, Fig.3). Wasserman (1978) argues that for low work intensities, exercise occurs under an isocapnic and isohydric state where increases in AV involved rises in both RR and TV. However during high-intensity exercise, anaerobic metabolism increases metabolic acidosis which prompts the condition of tachypnoea where RR increases to buffer the rising PaCO2 tension, bringing the system back into equilibrium (Wasserman, 1978). Thus the increase in RR under highintensity exercise is directly related to significant increases in AV confirming the hypothesis. The two-way ANOVA analysis found no statistical difference between baseline and end result in AV indicating a correct ventilatory recovery response (Fig. 1). Appropriate ventilation is defined in Whipp & Ward (1998) study as a function of pulmonary CO2 exchange, so it follows that AV will decrease during post-exercise recovery as CO2 stores move towards equilibrium. This idea is supported by the decreasing trend observed postexercise for AV, TV and RR (Fig. 1,2 &3). However studies report a neurogenic ventilatory reflex that is present during the abrupt changes from rest to exercise and also in the recovery

from exercise back to rest (Beaver & Wasserman, 1970). This neurogenic reflex is thought to result from the stimulation of proprioceptors and muscle spindles in exercising limbs (Beaver & Wasserman, 1970). This vagal reflex has been shown to alter breathing pattern increasing AV through increases in TV, supporting our results that found a significant rise in TV and not RR (Phillipson et al. 1970). However, contradicting studies into steady-state ventilation could not demonstrate that limb motion at a fixed work rate affected ventilation (Sipple & Gilbert, 1966). There are many competing theories into the mechanisms by which AV is controlled, however the most plausible mechanism for respiratory control during exercise was learned in the resection of carotid body chemoreceptors (Lugliani et al. 1971) Lugliani et al. (1971) study found an overall decrease in AV, RR and TV during exercise in the removal of carotid bodies and that metabolic acidosis failed to elicit the normal ventilatory response such that the degree of acidosis remained excessive. However the precise regulatory mechanism by which ventilation is able to couple the rate of CO2 delivery such that arterial CO2 tension and pH is tightly controlled still remains unknown (Wasserman, 1978). Wasserman (1978) findings split the ventilatory recovery response into three phases which dictate the pattern of change in minute ventilation. It was found that the induction of metabolic acidosis in phase II continued longer under heavy working intensities whilst lowintensity exercise reached an early plateau in three minutes as there was no production of lactic acid (Wasserman, 1978). However this trend is not reflected in the results as observed peak in AV for both low and high intensity exercise occurs immediately post-exercise at t = 10min and fall to resting at the same time interval at t = 15min (Fig. 1). Thus further investigation of the ventilatory response in comparing heavy and low working capacities should include a narrower timescale to allow the observation of trends immediately after exercise. The large variance observed in error bars at t = 10min (Fig. 1, 2 & 3) immediately postexercise can be attributed to the different levels at which metabolic acidosis is induced, depending on fitness or disorders that prevent oxygen transportation to muscle cells. Thus this study provides a method to examine the subjects physiological gas transport requirement to meet cellular oxygen demands. Results from this study may assist in clinical studies as exercise stress is commonly observed in congestive heart failure patients which usually suffer from unusual ventilatory response. Patterns in the defective ventilatory response underlying periodic breathing pertains relevant interest and should be investigated in future studies.

References Dempsey JA & Forster HV (1982). Mediation of ventilatory adaptations. Physiol Rev 62, 262346. Burton D A, Stokes K S, Hall GM (2004) Physiological Effects of Exercise. Journal of Anaesthesia vol. 4. No. 6. Pp.185-188 Phillipson EA, Hickey RF, Bainton CR et al: Effect of vagal blockade on regulation of breathing in conscious dogs. J Appl. Physiol 29:475-479, 1970 Lugliani R, Whipp BJ, Seard C, et al: Effect of bilateral carotid body resection of ventilatory control at rest and during exercise in man. N Engl J Med 285:1105-1111, 1971 Beaver & Wasserman 1970 Sipple J. H. and Gilbert R. Influence of proprioceptor activity in the ventilatory response to exercise. J. Appl. Physiol. 21: 143-146, 1966