Here we present a protocol, used to examine the effects on levels of biophysical and psychological stress following a visitation to three locations with different levels of nature. We're interested to learn whether a site with higher levels of naturalness will be more effective in helping participants to reduce their stress levels. In addition to using self report questionnaire, the main advantage of using this technique is that analyzing levels of cortisol and amylase collected from participants saliva samples can give us a more accurate read and better understanding of how our body actually responds after visitation to recreation sites.
Demonstrating the procedure will be Dr.Hocevar, a professor from Indiana University. Begin by introducing yourself to the subject. Provide them with a consent form to sign explaining the voluntary nature, purpose, and procedures of the study.
Give the subject a red armband for future identification. Obtain physiological and psychological measures of stress levels via saliva samples, just prior to the recreational experience, and immediately following conclusion of the experience. Collect a one to two milliliter saliva sample using a passive drool method.
Instruct subjects to allow saliva to pool in their mouths, then drool down the two inch straw and into the cryo vial until one milliliter has been collected. Label samples with an assigned three digit ID number and letter to indicate the timing of data collection. Then, temporarily freeze the sample in a steroid foam box full of dry ice for no more than two hours.
Transport the marked samples to a laboratory and store at minus 80 degrees Celsius until analyzed. Finally, use a liquid amylase reagent set to quantify alpha amylase in saliva samples. Then, use a multi mode reader capable of reading an optical density at 405 nanometers with the temperature controlled to 37 degrees Celsius during the assay.
First, acquire a cortisol enzyme amino acid kit to quantify cortisol in saliva samples, a spectrophotometer to read an optical density, or OD, at 450 nanometers as well as software capable of using OD recordings from the plate reader to perform four parameter logistic curve fitting. Allow all reagents to equilibrate to 20 to 25 degrees Celsius for a minimum of 30 minutes. Dilute the cortisol assay buffer one to five using deionized water, then dilute the wash buffer one to 20 using deionized water.
Next, thaw the samples on ice prior to analysis. Then, dilute samples one to 10 with cortisol assay buffer and use within two hours of preparation. To prepare the standards, label glass test tubes one through seven.
Then, pipet 225 microliters of assay buffer into tube number one, and 125 microliters of buffer into tubes two through seven. Next, add 25 microliters of the cortisol stock solution to tube number one, and vortex. Finally, remove 125 microliters of buffer from tube number one and add it to tube number two, then vortex again.
Results indicated a decrease in significant levels of salivary cortisol after visiting site A, the natural setting. No significant changes in levels of cortisol were observed in site B and C.Further, levels of alpha amylase increased after subjects visited site C.After visitation to the three locations and using the self report questionnaires, significant decreases on the factors of demands and worry were observed. No significant changes in the factor tension was observed for any of the three locations and significant increases were reported at sites A and site B for the factor joy.
Typically, much of the research done on stress reduction has relied on self report questionnaires. In this study, we collected participant's saliva samples to analyze how our body actually responds after visitation to three sites with different levels of naturalness. In this visualization, we featured both a laboratory techniques and one example of the in filled data collection.
This technique allows researcher to capture our body's reaction to different environments. In confirms our hypothesis that natural environment can have positive effects on human health. Future research is suggested to apply biophysical measurements of level of stress across different natural environments or different outdoor activities with true experimental design.
