Typically, rotenone-containing plants in the legume family, Fabaceae, are crushed and introduced into a body of water, and as rotenone interferes with cellular respiration, the affected fish rise to the surface in an attempt to gulp air, where they are more easily caught.

[16] When applied in freshwater systems, the treatment dose kills the target fish and usually other gilled species like tadpoles and zooplankton are affected, depending on dosage.

Its use is more benign for the environment (as compared to drying ponds, or using other piscicides), and studies show that most ecosystems naturally recover within one or two years after rotenone application- with aquatic invertebrates repopulating affected areas,[17][18][19] thus restoring initial local biodiversity to its status prior to the introduction of the invasive species.

[20] Norwegian authorities have been using rotenone since the mid-1980s to eradicate the salmon fluke Gyrodactylus salaris,[21] and as of 2024 48 out of 54 affected river catchments have been treated.

Additionally, many lakes and ponds have been rotenone treated in an effort to remove national or regional invasive species, such as Northern pike, roach, minnow, crucian carp, Tench and perch.

[22] In 1992, Florida Fish and Wildlife Conservation Commission (FWC) officials used rotenone to eradicate an established population of invasive jaguar cichlids from a small pond in Miami-Dade County.

In 2012, rotenone was used to kill all remaining fish in Stormy Lake (Alaska) due to invasive pike destroying native species, which were reintroduced once the treatment was concluded.

Rotenone was used in powdered form to treat scabies and head lice on humans, and parasitic mites on chickens, livestock, and pet animals.

[37] Rotenone works by interfering with the electron transport chain within complex I in mitochondria, which places it in IRAC MoA class 21 (by itself in 21B).

[41] Rotenone is produced by extraction from the roots and stems of several tropical and subtropical plant species, especially those belonging to the genera Lonchocarpus and Derris.

Rotenone was continuously applied over a period of five weeks, mixed with DMSO and PEG to enhance tissue penetration, and injected into the jugular vein.

Another study has also described toxic action of rotenone at low concentrations (5 nM) in dopaminergic neurons from acute rat brain slices.

[56] This toxicity was exacerbated by an additional cell stressor – elevated intracellular calcium concentration – adding support to the 'multiple hit hypothesis' of dopaminergic neuron death.

The neurotoxin MPTP had been known earlier to cause PD-like symptoms (in humans and other primates, though not in rats) by interfering with complex I in the electron transport chain and killing dopaminergic neurons in the substantia nigra.

However at least one study recently has found evidence of protein aggregation of the same chemical makeup as that which makes up Lewy bodies with similar pathology to Parkinson's disease in aged rhesus monkeys from MPTP.

In 2010, a study was published detailing the progression of Parkinson's-like symptoms in mice following chronic intragastric ingestion of low doses of rotenone.