The influence of the lunar cycle on bluefin tuna foraging in the upwelling area of the Strait of Messina was investigated by exploring trophic interaction with mesopelagic fish and cephalopod prey. To focus on how the lunar cycle could affect availability of mesopelagic prey for this predator, we tested potential differences in the diet related to each lunar phase. Moreover, we considered two potential impacts of the lunar cycle: the lunar irradiance and the strength of currents. Overall, 2672 prey items were mesopelagic fish and cephalopods, representing 60.7% of overall diet by number. The main mesopelagic fish prey items were lanternfishes and dragonfishes, while Onychoteuthis banksii was the most important cephalopod prey. In summary, the Strait of Messina has highly specific hydrodynamic and biological features which strongly depend on upwelling currents, which in turn are influenced by the lunar cycle (new and full moon with strong currents, quarters with fewer currents). Upwelling causes water mixing, bringing to the surface a large amount of mesopelagic fauna which become more readily available to tuna. Lunar irradiance contributes to the variation of prey composition, increasing the success of visual predation on mesopelagic resources at high light in the water column.