Effect of Coffee in Zebrafish

A research study on the exposure of Caffeine in zebrafish states that it can affect memory but may reduce stress. An overview of the research and the methods that have been carried out in the caffeine studies on zebrafish.

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Coffee, an infusion of ground, roasted coffee beans, is reported to be among the most widely consumed beverages in the world Although coffee is lauded for its aroma and flavor, its caffeine content likely plays a role in its popularity. Caffeine (1,3,7-trimethylxanthine) is a purine alkaloid that occurs naturally in coffee beans. At intake levels associated with coffee consumption, caffeine appears to exert most of its biological effects through the antagonism of the A1 and A2A subtypes of the adenosine receptor.

Mode of Induction:

  • The caffeine group was exposed to 20 mg/dL caffeine in group 1(P1), and 50mg/dL caffeine in group 2 (P2). 
  • A group with exposure to 20 mg/dL and 50 mg/dL caffeine extract administered the protocol toxicity in a range of 96 hours. 
  • The fish were initially held in a tank with two fish. After 96 hours of exposure to the caffeine group, P1 dan P2 was given a UCS. 
  • The other group was not assigned caffeine, the negative control group (K1) is only tested with the T-maze trial. 
  • Then, a positive control group (K2) was only given Unconditional Chronic Stress (UCS) for seven without caffeine.
Coffee image

Parameters to analyze neurotoxicity using behavior related assays:

1) T-maze

Fishes were individually transferred to T-maze and observed in one minute. Then, every group was noted for time and chosen arm color in T-maze as statistical analysis. There were two red and green arms. When fish got into the red component, being caught by a net for 30 seconds got punished. However, in the green arm, the Zebrafish got a reward of fish food. Furthermore, isolated in a green tank, the fish remembered the color. The training which was provided for the fish aims to increase neuroplasticity memory

2) Stress Exposure

Stress exposure was given to the K2, P1, and P2 groups for seven days. UCS was given to the fish which were different in a day. There was restrain stress which the centrifuge tube 4ml in 90 minutes. 

3) Predator Exposure 

This method was administered by putting the fish beside the predator with a divider. This method decreased water until the dorsal fin of the fish in two minutes. Chasing animals were applied for eight minutes using a chaser.

Moreover, the tank water replacement method was performed in changing the zebrafish tank with new water three times when the zebra must adapt in freshwater. UCS exposure is the same in every group to prevent the different outcome of stress.


Caffeine affects memory and stress. Caffeine may reduce stress at a low dose. Moreover, caffeine affects the memory in Zebrafish.


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