Effects of different chilling procedures on honey bees (Apis mellifera) for anesthesia

Abstract

Immobilization is required for the examination and manipulation of honey bees (Apis mellifera), and many techniques have been developed to render honey bees immobile until this date. Among them, two methods, which are carbon dioxide (CO2) and low-temperature narcosis, have been commonly used to induce anesthesia on the bees. Although CO2-induced anesthesia changes the behavior and physiology of honey bees, the anesthesia induced by cold is considered commonly as non-damaging for the bees. There are a very limited number of studies on the effects of different chilling protocols in honey bees. Therefore, the purpose of this study was to investigate the effect of different chilling procedures on the survival rate and depth of anesthesia in honey bees. A total of 4 groups were formed, one of which was the control group. Three temperatures (+4°C, -20°C and -80°C) were applied to three different experimental groups. Furthermore, four different exposure times were implemented on each experimental group (n=20 worker bees in each trial). The depth of anesthesia was assessed and scored based on the movements and anesthesia recovery time in honeybees. There was a statistically significant relationship between duration and survival/death rate in the applications at +4°C, -20°C and -80°C (P<0.001). The method that was applied for 5 minutes at -20°C was the most ideal chilling method (P<0.03) and showed the lowest mortality in addition to the high depth of anesthesia. This study may assist in selecting the safest and deepest anesthesia method required in any study on honey bees.

Keywords

• Anesthesia,chilling,honeybee

References

1. 1. Austin GH (1955): Effect of carbon dioxide anaesthesia on bee behaviour and expectation of life. Bee World, 36, 45-47.
2. 2. Becher MA, Scharpenberg H, Moritz RFA (2009): Pupal developmental temperature and behavioral specialization of honeybee workers (Apismellifera L.). J Comp Physiol A, 195, 673-679.
3. 3. Bogdanov S, Jurendic T, Sieber R, et al (2008): Honey for nutrition and health: a review. J Am Coll Nutr, 27, 677-689.
4. 4. Bühler A, Lanzrein B, Wille H (1983): Influence of temperature and carbon dioxide concentration on juvenile hormone titre and dependent parameters of adult worker honey bees (Apismellifera L.). J Insect Physiol. 29, 885-893.
5. 5. Cooper JE (2001): Invertebrate anesthesia. Vet Clin North Am Exot Anim Pract, 4, 57-67.
6. 6. Cooper JE (2011): Anesthesia, analgesia, and euthanasia of invertebrates. ILAR J, 52, 196-204.
7. 7. Czekońska K (2009): The effect of different concentrations of carbon dioxide (CO2) in a mixture with air or nitrogen upon the survival of the honey bee (Apismellifera). J Apic Res, 48, 67-71.
8. 8. Ebadi R, Gary NE, Lorenzen K (1980): Effects of carbon dioxide and low temperature narcosis on honey bees, Apismellifera. Environ Entomol, 9, 144-150.

9. 9. Engels W, Ramamurty RS (1976): Initiation of oogenesis in allatectomised virgin honey bee queens by carbon dioxide treatment. J Insect Physiol, 22, 1427-1432.
10. 10. Eskov EK, Eskova MD, Spasik SE (2013): Changes in body mass and life span of honeybee Apismellifera L. under effect of narcotization with carbon dioxide. J Evol Biochem Physiol, 49, 627-629.
11. 11. Frost EH, Shutler D, Hillier NK (2011): Effects of cold immobilization and recovery period on honeybee learning, memory, and responsiveness to sucrose. J Insect Physiol, 57, 1385-1390.
12. 12. Groening J, Venini D, Srinivasan MV (2018): Effects of cold anaesthesia on the defensive behaviour of honeybees. Insect Soc, 65, 359-366.
13. 13. Harris JW, Woodring J (1995): Elevated brain dopamine levels associated with ovary development in queenless worker honey bees (Apismellifera L.). Comp Biochem Physiol C Pharmacol Toxicol Endocrinol, 111, 271-279.
14. 14. Harris JW, Woodring J, Harbo JR (1996): Effects of carbon dioxide on levels of biogenic amines in the brains of queenless worker and virgin queen honey bees (Apis mellifera). J Apic Res, 35, 69-78.
15. 15. Human H, Brodschneider R, Dietemann V, et al (2013): Miscellaneous standard methods for Apis mellifera research. J Apic Res, 52, 1-53.
16. 16. Jones CJ, Helliwell P, Beekman M, et al (2005): The effects of rearing temperature on developmental stability and learning and memory in the honeybee, Apis mellifera. J Comp Physiol A,191, 1121-1129.
17. 17. Klein AM, Vaissiere BE, Cane JH, et al (2007): Importance of pollinators in changing landscapes for world crops. Proc R Soc B, 274, 303-313.
18. 18. Koywiwattrakul P, Thompson GJ, Sitthipraneed S, et al (2005): Effects of carbon dioxide narcosis on ovary activation and gene expression in worker honeybees, Apis mellifera. J Insect Sci, 5, 1-10.
19. 19. Madras-Majewska B, Kaminski Z, Balcerak M, et al (2013): Effect of different oxygen and nitrogen ratios in bees awakened from carbon dioxide anesthesia on their behavior and survival after this exertion. Med Weter, 69, 760-762
20. 20. Neumann P, Carreck NL (2010): Honey bee colony losses. J Apic Res, 49, 1-6.
21. 21. Nicolas G, Sillans D (1989): Immediate and latent effects of carbon dioxide on insects. Annu Rev Entomol, 34, 97-116.
22. 22. Olszewski K, Borsuk G, Paleolog, et al (2012): Influence of carbon dioxide anaesthesia on the length of worker life and food foraging in cage tests. Med Weter, 68, 615-617
23. 23. Ribbands CR (1950): Changes in the behaviour of honey-bees following their recovery from anaesthesia. J Exp Biol, 27, 302-310.
24. 24. Robinson GE, Visscher PK (1984): Effect of low temperature narcosis on honey bee (Hymenoptera: Apidae) foraging behaviour. Fla Entomol, 67, 568–570.
25. 25. Southwick EE (1985): Allometric relations, metabolism and heart conductance in clusters of honey bees at cool temperatures. J Comp Physiol B, 156, 143-149.
26. 26. Stabentheiner A, Kovac H, Brodschneider R (2010): Honeybee colony thermoregulation–regulatory mechanisms and contribution of individuals in dependence on age, location and thermal stress. PLoS One, 5, e8967.
27. 27. Tustain RCR, Faulke J (1979): Effect of carbon dioxide anaesthesia on the longevity of honey bees in the laboratory. New Zeal J Exp Agr, 7, 327-329.
28. 28. Tutun H, Koç N, Kart A (2018): Plant essential oils used against some bee diseases. Turkish JAF Sci Tech, 6, 34-45.
29. 29. Wilde J, Sobiechowski K (2002): The effect of carbon dioxide treatment on matinf of virgin honeybee queens. Pol J Natur Sci, 12, 73-85.
30. 30. Woyke J, Fliszkiewicz C, Jasiñski Z (2001): Prevention of natural mating of instrumentally inseminated queen honeybees by proper method of instrumental insemination. J Apic Sci, 45, 101-114.