Agitation is one of the hidden values all slimes have which determines which standard facial expression slimes assume in relation to their agitation and hunger values, and is the primary trigger for how often their negative behaviors occur. It takes roughly 48 in-game hours for a fully fed happy slime to reach 100% agitation without modifiers.
Slimes are set to Elated within The Slimeulation, and their agitation does not decay.
Agitation naturally increases at all times, similarly to hunger. There are outside modifiers which can affect the rate of agitation in various ways;
Feeding a slime is the only way to completely lower agitation permanently and restore it to a happy state. However, it can only eat when it is hungry. Feeding a slime its favorite food will reduce its agitation by 30%. Feeding it anything else, with the exception of tofu, will reduce agitation by 15%. Tofu reduces agitation by 1.25-1.3% 
Water and Ancient WaterWater and Ancient Water can negate negative behaviors and reduces agitation by 10% for 20 seconds. This decrease does not stack and is temporary, but may be reapplied after the 20 seconds are up.
The Music Box is an upgrade to the Corral which halves natural agitation growth.
Slime ToysSlime Toys are purchasable items unlocked through the 7Zee Rewards Club which reduces the natural rate of agitation growth by 12.5%, or 25% if any toy in range is favored by the slimes. Additional Slime Toys do not stack their effects, but can stack with a Music Box.
Tangle Slime PollenTangle Slimes produce a pollen aura which doubles the natural rate of agitation growth. Multiple pollen auras do not stack this effect. Other Tangle Slimes are immune to this effect.
Fire Slime ContactFire Slimes on fire can severely agitate slimes on contact by approximately 33.33% temporarily. Any agitation caused by Fire Slime contact will take up to three in-game hours to completely wear off.
Other Fire Slimes are immune to this effect.
Certain slimes have negative behaviors which are performed much more frequently or made apparent once they reach a certain agitation state. Many of these behaviors can be suppressed for 20 seconds by splashing the slime with Water or Ancient Water. This includes:
- Rock Slimes roll every 15 seconds at 0% agitation. Rolling frequency is increased at higher agitation, which is capped to every 3 seconds at 100% agitation.
- Rad Slimes expand their aura every 180 seconds at 0% agitation. Expansion frequency is increased at higher agitation, which is capped to every 30 seconds at 100% agitation.
- Boom Slimes explode every 45 seconds at 0% agitation. Explosion frequency is increased at higher agitation, which is capped to every 10 seconds at 100% agitation.
- Crystal Slime crystal production occurs every 15 seconds at 0% agitation. Crystal production frequency is increased at higher agitation, which is capped to every 3 seconds at 100% agitation.
- Hunter Slime Largos can go feral at 99.99% agitation.
- Quantum Slimes produce ghosts, or qubits, every 20 seconds at 0% agitation. Qubit production is increased at higher agitation, which is capped to every 5 seconds at 100% agitation. They begin swapping places with their qubits at 20% agitation.
- Mosaic Slimes produce glints every 30 seconds at 0% agitation. Glint production frequency is increased at higher agitation, which is capped to every 6 seconds at 100% agitation. The range of production is increased at 95%.
- Dervish Slimes will produce large tornados at 95% agitation.
- Tangle Slimes produce pollen which causes nearby slimes to sneeze, agitating them. They begin doing this at 75% agitation.
- Gold Slimes and Lucky Slimes are susceptable to hunger and agitation like any other slime. However, Lucky Slimes do not react to becoming agitated or starving. Due to their rarity and despawning mechanics, this does not normally come into play under normal circumstances.
- Tarr are also subjected to agitation, although are set to 100% every frame.
- Puddle Slimes can not naturally get above 20% permanent Agitation without tangle pollen. They would evaporate before this happened.
- Although slime agitation is at a set level within The Slimeulation to always be Elated, Fire Slimes are still capable of temporarily agitating them. However this does not occur under normal circumstances since Fire Slimes cannot be found there.