Effect of TFM on stress and metabolism in rainbow trout

In 2018, researchers Oana Birceanu and Michael Patrick Willie explored the effect of a
pesticide called TFM on rainbow trout, a fish not targeted by the pesticide. Historically, it
has been used to prevent overgrowth of invasive sea lamprey populations in the Great
Lakes. TFM is toxic to sea lampreys due to its ability to stop the mitochondria from
properly producing ATP, which can eventually lead to death. Rainbow trout and other
non-target fishes are not as sensitive to the chemical due to a better ability to detoxify
the chemical. Researchers measured both the stress and metabolic response to TFM
exposure, as well as their ability to handle other acute stressors using 2 main
techniques: in vivo, which involves the live organisms, and in vitro, which involves cell
culture in a petri dish. Overall, researchers found that the metabolic capacity and ability
to produce cortisol- a major stress hormone- in the rainbow trout remained
uncompromised following TFM exposure, indicating no long-term effect on their ability to
cope with stressors.

In order to investigate the effects of in vivo TFM exposure, researchers divided the trout
into two groups: one group exposed to the minimum amount of TFM needed to kill the
sea lampreys (minimum lethal concentration)- 7.6 mg/L- for 9 hours, and the other
group served as controls and were not exposed to TFM. 9 hours was chosen because it
mimics the amount of time non-target fishes would typically be exposed during a TFM
treatment. A drip-set using a pump allowed researchers to maintain a constant TFM
drip.12 hours after the TFM drip was stopped, researchers removed some trout from
both the control and experimental group, and euthanized them. The remaining trout
were exposed to an acute stressor and had their blood and tissues sampled 1, 4, and
24 hours post-stress. Researchers also investigated the effects of in vitro TFM exposure
by first dividing juvenile rainbow trout into a control group and group exposed to TFM in
similar conditions as in the in vivo experiment. Researchers then euthanized the fish
and removed and preserved their head-kidneys. The livers were minced, divided into
wells, washed, incubated, and placed on dry ice.

An enzyme-linked immunosorbent assay was used to measure cortisol levels from both
in vivo and in vitro approaches. A glucose assay was used to colorimetrically measure
glucose levels, and liver protein was determined using a spectrophotometer. Lactate
levels were determined using an enzyme. Changes in glucose levels were used to
measure glycogen levels in the liver, and a spectrophotometer was used to then
analyze the liver’s metabolic capacity. Gene analysis was performed on RNA taken from
the head-kidney.

Overall, researchers found significantly higher cortisol concentrations in TFM-exposed

fish compared to non-exposed controls. Glucose levels in the TFM group also remained
elevated (1.5 fold higher) compared to the control group. Researchers found no major
effects of prior TFM exposure on gene abundance compared to controls. However, for
the in vitro cortisol production and glycogen stores were not significantly different
between TFM-exposed individuals and controls.