# Green Veterinary Pharmacology Applied to Beekeeping: Semi-Field and Field Tests Against Varroa destructor, Using Essential Oil of Bergamot (Citrus bergamia) and Lemon (Citrus limon)

## TL;DR

This study tests the effectiveness of bergamot and lemon essential oils in reducing Varroa mite infestations in honeybees under semi-field and field conditions.

## Contribution

The study evaluates the acaricidal efficacy of two essential oils in real-world beekeeping settings, highlighting the need for controlled-release formulations.

## Key findings

- Bergamot and lemon essential oils achieved 40.3% and 50.7% Varroa mite neutralization at the highest field concentrations.
- Essential oils showed no toxicity to honeybees in semi-field tests.
- Field acaricidal efficacy was lower than in laboratory tests, suggesting the need for improved formulations.

## Abstract

Varroa destructor parasitosis represents the most serious threat to honeybee health. The parasite causes damage to the host, which undermines its survival. Controlling infestation levels through drug treatment becomes a critical process for a colony. In this study, we verified the acaricidal efficacy of two essential oils (Citrus limon and Citrus bergamia) that, in previous laboratory tests performed by residual contact toxicity tests, have shown good toxicity toward Varroa. Bergamot and Lemon essential oils returned, at the highest concentration used in the field, an average percentage of Varroa neutralization of 40.3 and 50.7%, respectively. Advanced formulations that allow the gradual and controlled release of essential oil over time are needed to increase acaricidal efficacy.

Varroa destructor is an ectoparasitic mite that causes a true syndrome with complex symptoms in honeybees. For mite control, several synthetic active principles are commercially available. However, these formulations are proving less effective due to the acquisition of resistance mechanisms by the parasite. Naturally derived compounds, such as essential oils, are less prone to generating resistance mechanisms in mite populations and have been proven effective in numerous independent laboratory investigations. In this study, semi-field and field tests were conducted to evaluate the acaricidal efficacy of two essential oils (Citrus bergamia and Citrus limon), which, in our previous residual contact toxicity tests, showed efficacy against Varroa mites. For semi-field cage tests, two-level cages were set up. Filter paper soaked with different concentrations of essential oils was placed in the lower compartment of the cage; 20 honeybees and 10 Varroa mites were allocated in the upper level. Mite detachment from the honeybees was evaluated after 24 and 48 h. For field tests, cardboard strips soaked with solutions of essential oils at different concentrations were inserted in the experimental hives. Weekly, the strips were replaced, and the fall of parasites on the diagnostic bottom board was assessed. Semi-field tests showed that the essential oils, at the concentrations used, were not toxic to bees. In the semi-field tests, an average acaricidal efficacy of 33% and 60% was obtained, at the highest concentration used, for bergamot and lemon essential oils, respectively. During the 4 weeks of exposure to 600 mL of BEO, the average mite mortality was 28.5%. In the experimental group treated with the intermediate concentration of 1200 mL, the average mite mortality at the end of four weeks was 30.2%, while with the highest concentration of 1800 mL, it was 40.3%. During the four weeks of exposure to 600 mL of LEO, the average mite mortality was 34.1%. In the experimental group treated with the 1200 mL concentration, the average mite mortality at the end of the four weeks was 46.6%, while with the highest concentration, it was 50.7%. This study shows that the acaricidal efficacy observed in field tests differs from that in laboratory tests. Innovative formulations are needed that allow the time-controlled and gradual release of essential oil in the field.