Although the actual questions are missing from your ask, here's how you can tell.
If the study will be messed up (added bias) through interaction with the subjects the study should be observational. For example if a biologists wanted to study the eating habits of wild birds in the tropics, they would need to do an observational study. if they did a controlled experiment, the birds will change their eating habits based on human interaction, thus making the study invalid.
If the study requires specific measurements, close monitoring, or contact and interaction with the subjects the study should be a controlled experiment. For example, if a scientist wanted to test out a new drug, the health of patients would need to be closely monitored.
Answer:
I think it's A) A protein substrate
I believe the biosphere is fragile, because over time all of the pollution us humans have made; If its from air pulloution, acid rain or even water pollution. the toxic acids over time have made the biosphere and the ozon layer weaker and more thin. which causes the biosphere to become weaker.