We have found that giant hornets (Vespa mandarinia japonica) are killed in less than 10 min when they are trapped in a bee ball created by the Japanese honeybees Apis cerana japonica, but their death cannot be solely accounted for by the elevated temperature in the bee ball. In controlled experiments, hornets can survive for 10 min at the temperature up to 47 degrees C, whereas the temperature inside the bee balls does not rise higher than 45.9 degrees C. We have found here that the CO2 concentration inside the bee ball also reaches a maximum (3.6 +/- 0.2%) in the initial 0-5 min phase after bee ball formation. The lethal temperature of the hornet (45-46 degrees C) under conditions of CO2 concentration (3.7 +/- 0.44%) produced using human expiratory air is almost the same as that in the bee ball. The lethal temperature of the honeybee is 50-51 degrees C under the same air conditions. We concluded that CO2 produced inside the bee ball by honeybees is a major factor together with the temperature involved in defense against giant hornets.
Upon capture in a bee ball (i.e., a dense cluster of Japanese honeybees forms in response to a predatory attack), an Asian giant hornet causes a rapid increase in temperature, carbon dioxide (CO₂), and humidity. Within five min after capture, the temperature reaches 46°C, and the CO₂ concentration reaches 4%. Relative humidity gradually rises to 90% or above in 3 to 4 min. The hornet dies within 10 min of its capture in the bee ball. To investigate the effect of temperature, CO₂, and humidity on hornet mortality, we determined the lethal temperature of hornets exposed for 10 min to different humidity and CO₂/O₂ (oxygen) levels. In expiratory air (3.7% CO₂), the lethal temperature was ≥ 2° lower than that in normal air. The four hornet species used in this experiment died at 44-46°C under these conditions. Hornet death at low temperatures results from an increase in CO₂ level in bee balls. Japanese honeybees generate heat by intense respiration, as an overwintering strategy, which produces a high CO₂ and humidity environment and maintains a tighter bee ball. European honeybees are usually killed in the habitat of hornets. In contrast, Japanese honeybees kill hornets without sacrificing themselves by using heat and respiration by-products and forming tight bee balls.
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