Fermentation of a Pollen Substitute Diet with Beebread Microorganisms Increases Diet Consumption and Hemolymph Protein Levels of Honey Bees (Hymenoptera, Apidae)

Joyce Mayra Volpini Almeida Dias, Michelle Manfrini Morais, Thiago Mauricio Francoy, Rogério Aparecido Pereira, Aline Patricia Turcatto, David de Jong


Pollen substitute diets have become increasingly important for maintaining strong and healthy honey bee colonies. Palatability and nutritional value are key attributes of a good diet. Since beebread, which is pollen fermented by the bees, is the main food of the worker nurse bees that feed and care for the bee larvae, pollen substitutes should have similar attributes. In an attempt to simulate this natural food source, an inoculum prepared from beebread was used to ferment a pollen-substitute diet. Newly emerged bees were fed on the diets for seven days. They consumed significantly more fermented than unfermented diet. Hemolymph protein levels were significantly higher in bees that had been fed a fermented versus an unfermented diet, though still significantly lower than in bees fed on beebread. Vitellogenin (a key storage protein for honey bees) levels were also increased significantly in bees fed the fermented versus the non-fermented diet. Survival rates were higher for bees fed the fermented versus the non-fermented diet, though the difference was not significant. We conclude that fermentation by beebread-derived microorganisms can improve the acceptance and utility of an artificial protein diet for honey bees.


Apis mellifera, Africanized honey bees, fermented, artificial diet.

Full Text:



Abbas, T., Abid, H. & Ali, R. (1995). Black gram as a pollen substitute for honey bees. Animal Feed Science and Technology, 54: 357-359. doi: 10.1016/0377-8401(95)00772-F

Anderson, K.E., Carroll, M.J., Sheehan, T., Mott, B.M., Maes, P. & Corby-Harris, V. (2014). Hive-stored pollen of honey bees: many lines of evidence are consistent with pollen preservation, not nutrient conversion. Molecular Ecology, 23: 5904-5917. doi: 10.1111/mec.12966

Barragan, S., Basualdo, M. & Rodriguéz, E.M. (2016). Conversion of protein from supplements into protein of hemolymph and fat bodies in worker honey bees (Apis mellifera L). Journal of Apicultural Research, 54: 399-404. doi: 10.1080/00218839.2016.1158534

Basualdo, M., Barragan, S., Vanagas, L., Garcia, C., Solana, H., Rodriguez, E. & Bedascarrasbure, E. (2013). Conversion of high and low pollen protein diets into protein in worker honey bees (Hymenoptera: Apidae). Journal of Economic Entomology, 106: 1553-1558. doi: 10.1603/EC12466

Basualdo, M., Barragan, S. & Antunez, K. (2014). Bee bread increases honeybee haemolymph protein and promote better survival despite causing higher Nosema ceranae abundance in honeybees. Environmental Microbiology Reports, 6: 396-400. doi: 10.1111/1758-2229.12169

Bitondi, M.M. G. & Simões, Z.L.P. (1996). The relationship between level of pollen in the diet, vitellogenin and juvenile hormone titers in africanized Apis mellifera workers. Journal of Apicultural Research, 35: 27-36. doi: 10.1080/00218839.1996.11100910

Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72: 248-254.

Brodschneider, R., & Crailsheim, K. (2010). Nutrition and health in honey bees. Apidologie, 41: 278–294. doi: 0.1051/apido/2010012

Cappelari, F.A., Turcatto, A.P., Morais, M.M., & De Jong, D. (2009). Africanized honey bees more efficiently convert protein diets into hemolymph protein than do Carniolan bees (Apis mellifera carnica). Genetics and Molecular Research, 8: 1245-1249. doi: 10.4238/vol8-4gmr628

Carroll, M.J., Brown, N., Goodall, C., Downs, A.M., Sheenan, T.H. & Anderson, K.E. (2017). Honey bees preferentially consume freshly-stored pollen. Plos One, 12(4): e0175933. doi: 10.1371/journal.pone.0175933

Crailsheim, K. (1990). The protein balance of the honey bee worker. Apidologie, 21: 417–429.

Cremonez, T.M., De Jong, D. & Bitondi, M.M.G. (1998). Quantification of hemolymph proteins as a fast method for testing protein diets for honey bees (Hymenoptera: Apidae). J Journal of Economic Entomology, 91: 1284-1289. doi: 10.1093/jee/91.6.1284

DeGrandi-Hoffman, G., Chen, Y., Huang, E. & Huang, M.H. (2010). The effect of diet on protein concentration, hypopharyngeal gland development and virus load in worker honey bees (Apis mellifera L.). Journal of Insect Physiology, 56: 1184–1191. doi: 10.1016/j.jinsphys.2010.03.017

DeGrandi-Hoffman, G., Eckholm, B.J. & Huang, M. H. (2013). A comparison of bee bread made by Africanized and European honey bees (Apis mellifera) and its effects on hemolymph protein titers. Apidologie, 44: 52-63. doi: 10.1007/s13592-012-0154-9

DeGrandi-Hoffman, G., Chen, Y.P., Rivera, R., Carroll, M., Chambers, M., Hidalgo, G. & de Jong, E.W. (2016). Honey bee colonies provided with natural forage have lower pathogen loads and higher overwinter survival than those fed protein supplements. Apidologie, 47: 186-196. doi: 10.1007/s13592-015-0386-6

De Jong, D., Silva, E.J., Kevan, P. & Atkinson, J.L. (2009). Pollen substitutes increase honey bee hemolymph protein levels as much as or more than does pollen. Journal of Apicultural Research, 48: 34-37. doi: 10.3896/IBRA.

Dietz, A. & Stevenson, H.R. (1980). Infl uence of long term storage on the nutritional value of frozen pollen for brood rearing of honey bees, Apidologie: 11, 143-151. doi: 10.1051/apido:19800204

Ellis, A. & Hayes, G.W. (2009). An evaluation of fresh versus fermented diets for honey bees (Apis mellifera). Journal of Apicultural Research, 48: 215-216. doi: 10.3896/IBRA.

Gilliam, M. (1997). Identification and roles of non-pathogenic microflora associated with honey bees. FEMS Microbiology Letters, 155: 1-10. doi: 10.1111/j.1574-6968.1997.tb12678.x

Gilliam, M., Prest, D. & Lorenz, B. (1989). Microbiology of pollen and bee bread: taxonomy and enzymology of molds. Apidologie, 20: 53–68. doi: 10.1051/apido:19890106

Hagedorn, H.H. & Moeller, F.E. (1968). Effect of the age of pollen used in pollen supplements on their nutritive value for the honeybee. I. Effect on thoracic weight, development of hypopharyngeal glands, and brood rearing. Journal of Apicultural Research, 7: 89-95. doi: 10.1080/00218839.1968.11100195

Herbert, E.W., Shimanuki, H. & Caron, D. (1977). Optimum protein levels required by honey bees (Hymenoptera: Apidae) to initiate and maintain brood rearing. Apidologie, 8: 141-146. doi: 10.1051/apido:19770204

Herbert, E.W. & Shimanuki, H. (1978). Chemical composition and nutritive value of bee collected and bee stored pollen. Apidologie. 9: 33-40. doi: 10.1051/apido:19780103

Hoover, S.E.R., Higo, H.A. & Winston, M.L. (2006). Worker honey bee ovarian development: seasonal variation and the influence of larval and adult nutrition. Journal of Comparative Physiology B, 176: 55–63. doi: 10.1007/s00360-005-0032-0

Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage t4. Nature. 227: 680-685. doi: 10.1038/227680a0

Loper, G., Standifer, L., Thompson, M. & Gilliam, M. (1980). Biochemistry and Microbiology of bee-collected almond (Prunus dulcis ) pollen and beebread. Apidologie, 11: 63–73. doi: 10.1051/apido:1980010

Maes, P.W., Rodrigues, P.A.P., Oliver, R., Mott, B.M. & Anderson, K. E. (2016). Diet-related gut bacterial dysbiosis correlates with impaired development, increased mortality and Nosema disease in the honeybee (Apis mellifera). Molecular Ecology, 25: 5439-5450. doi: 10.1111/mec.13862

Manning, R., Rutkay, A., Eaton, L. & Bernard, D. (2007). Lipid-enhanced pollen and lipid-reduced flour diets and their effect on the longevity of honey bees (Apis mellifera L.). Australian Journal of Entomology, 46: 251–257. doi: 10.1111/j.1440-6055.2007.00598.x

Mattila, H.R. & Otis, G. W. (2006). Effects of pollen availability and Nosema infection during the spring on division of labor and survival of worker honey bees (Hymenoptera: Apidae). Environmental Entomology, 35: 708-717. doi: 10.1603/0046-225X-35.3.708

Morais, M.M., Turcatto, A.P., Francoy, T.M., Gonçalves, L.S., Cappelari, F.A. & De Jong, D. (2013a). Evaluation of inexpensive pollen substitute diets through quantification of haemolymph proteins. Journal of Apicultural Research, 52: 119-121. doi: 10.3896/IBRA.

Morais, M.M., Turcatto, A.P., Pereira, R.A., Francoy, T.M., Guidugli-Lazzarini, K.R., Gonçalves, L.S., Almeida, J.M.V., Ellis, J.D. & De Jong, D. (2013b). Protein levels and colony development of Africanized and European honey bees fed natural and artificial diets. Genetics and Molecular Research, 12: 6915-6922. doi: 10.4238/2013

Paiva J.P.L.M., Paiva, H.M., Esposito, E. & Morais, M.M. (2016). On the effects of artificial feeding on bee colony dynamics: a mathematical model. Plos One, 11(11): e0167054. doi:10.1371/ journal.pone.0167054.

Pernal, S.F. & Currie, R.W. (2002). Discrimination and preferences for pollen-based cues by foraging honeybees, Apis mellifera L. Animal Behavior, 63: 369-390. doi: 10.1006/anbe.2001.1904

Robinson, F. & Nation, J. (1968). Substances that attract caged honeybee colonies to consume pollen supplements and substitutes. Journal of Apicultural Research, 7: 83–88. doi: 10.1080/00218839.1968.11100194

Roulston, T.H. & Cane, H.J. (2000). Pollen nutritional content and digestibility for animals. Plant Systematics and Evolution, 222: 187-209.

Rousseau, A. & Giovenazzo, P. (2016). Optimizing drone fertility with spring nutritional supplements to honey bee (Hymenoptera: Apidae) colonies. Journal of Economic Entomology, 109: 1009-1014. doi: 10.1093/jee/tow056

Schmidt, J.O. & Hanna, A. (2006). Chemical nature of phagostimulants in pollen attractive to honeybees. Journal of Insect Behavior, 19: 521-532. doi: 10.1007/s10905-006-9039-y

Schmidt, J.O., Thoenes, S.C. & Levin, M.D. (1987). Survival of honey bees, Apis mellifera (Hymenoptera: Apidae), fed various pollen sources. Journal of Economic Entomoloogy, 80: 176-183. doi: 10.1093/aesa/80.2.176

Somerville, D. (2005). Fat bees skinny bees—a manual on honey bee nutrition for beekeepers. Rural Industries Research and Development Corporation, Barton.

Standifer, L.N., Haydak, M.H., Mills, J.P. & Levin, M.D. (1973). Value of three protein rations in maintaining honey bee colonies in outdoor flight cages. Journal of Apicultural Research, 12: 137-143.

Winston, M.L., Chalmers, W.T. & Lee, P.C. (1983). Effects of two pollen substitutes on brood mortality and length of adult life in the honey bee. Journal of Apicultural Research, 22: 49-52.

van der Steen, J. (2007). Effect of a home-made pollen substitute on honey bee colony development. Journal of Apicultural Research, 46: 114-119. doi: 10.1080/00218839.2007.11101377

Vasquez, A. & Olofsson, T. (2009). The lactic acid bacteria involved in the production of bee pollen and bee bread. Journal of Apicultural Research, 48: 189–195. doi: 10.3896/IBRA.

DOI: http://dx.doi.org/10.13102/sociobiology.v65i4.3293


  • There are currently no refbacks.

JCR Impact Factor 2016: 0.699