Assessing Sperm Quality in Stingless Bees


  • Hiara Marques Meneses Federal University of Ceará
  • Sheina Koffler University of São Paulo
  • Breno Magalhaes Freitas Federal University of Ceará
  • Vera Lucia Imperatriz-Fonseca University of São Paulo
  • Rodolfo Jaffé University of São Paulo



sperm viability, sperm morphology, sperm counts, semen


Although stingless bees have a great potential as commercial pollinators, their exploitation depends on the successful reproduction of colonies on a large scale. To do so, it is essential to develop accurate diagnostic tools that facilitate a better understanding of the reproductive biology of stingless bees. Sperm counts, sperm morphology and sperm viability (the relative proportion of live to dead sperm), are key parameters assessing semen quality and potential fertilization success. Here we present standardized protocols to assess these three parameters. We used Scaptotrigona aff. depilis (Moure) as a study model. Semen extractions from the seminal vesicles were found to yield better results when performed in mature rather than in younger males. For morphology and viability analyses, the best semen dilution on Hayes solution was adding 120 µl to the contents of the two seminal vesicles. For sperm count, however, we recommend a higher dilution (1.000 µl). Sperm viability values were higher when Hayes solution was adjusted to pH 8.7 and no difference were found in sperm viability when analyzing fresh semen, and semen collected 1, 3 and 5 hours prior to the analyzes. However, we found a significant decrease in sperm viability when analyzing sperm collected 24 hours beforehand. The protocols seem sensitive and efficient, and show great potential to be used in other stingless bee species.


Ball, B. A., Medina, V., Gravance, C. G. & Baumber, J. (2001). Effect of antioxidants on preservation of motility, viability and acrosomal integrity of equine spermatozoa during storage at 5ºC. Theriogenology 56: 577-589.

Camillo, C. (1971). Estudos adicionais sobre os zangões de Trigona (friesella) schrottkyi (Hym. Apidae). Ciência e Cultura 29:279.

Collins, A. M. & Donoghue, A. M. (1999). Viability assessment of honey bee, Apis mellifera sperm using dual fluorescent staing. Theriogenology 51: 1513-1523.

Collins, A. M. (2004). Sources of variation in the viability of honey bee, Apis mellifera L., semen collected for artificial insemination. Invertebr Reprod Dev 45:231-237

Cortopassi-Laurino, M. (1979). Observações sobre atividade de machos de Plebeia droryana friese (Apidae, Meliponinae). Rev Bras Entomol 23:177-191.

den Boer, S. P. A., Boomsma, J. J. & Baer, B. (2009). Honey bee males and queens use glandular secretions to enhance sperm viability before and after storage. J Insect Physiol 55:538-543.

Ferreira, A., Abdalla, F. C., Kerr, W. E. & Cruz-Landim, C. (2004). Comparative anatomy of the male reproductive internal organs of 51 species of bees. Neotropical Entomol 33:569-576.

Garner, D. L., Johnson, L. A., Yue, S. T., Roth, B. L. & Haugland, R. P. (1994). Dual DNA staining assessment of bovine sperm viability using SYBR-14 and propidium iodide. J Androl 15: 620-629.

Garófalo, C. A. (1980). Reproductive aspect and evolution of social behavior in bees (Hymenoptera, Apoidea). Braz J Genet 3:139-152.

Gençer, H. V.; Kahya, Y. & Woyke J. (2014). Why the viability of spermatozoa diminishes in the honeybee (apis mellifera) within short time during natural mating and preparation for instrumental insemination. Apidologie doi:10.1007/s13592-014-0295-0.

Holman, L. (2009). Sperm vianility staining in ecology and evolution: potential pitfalls. Behav Ecol Sociobiol 63. 11: 1679-1688.

Paulenz, H., Soderquist, L., Pérez-Pé, R. & Berg, K. A. (2002). effect of different extenders and storage temperatures on sperm viability of liquid ram semen. Theriognology 57: 823-836.

Pech-May, F. G., Medina-Medina, L., May-Itzá, W. J., Paxton, R. J. & Quezada-Euán, J. J. G. (2012). Colony pollen reserves affect body size, sperm production and sexual development in males of the stingless bee Melipona beecheii. Insect Soc 59:417-424.

Rzymski, P., Langowska, A., Fliszkiewicz, M. , Poniedzialek, B., Karczewski, J. & Wiktorowicz, K. (2012). Flow cytometry as an estimation tool for honey bee sperm viability. Theriogenology 77:1642-1647.

Simmons, L. W. (2001). Sperm competition and its evolutionary consequences in the insects. Princeton Univ. Press, Priceton.




How to Cite

Meneses, H. M., Koffler, S., Freitas, B. M., Imperatriz-Fonseca, V. L., & Jaffé, R. (2014). Assessing Sperm Quality in Stingless Bees. Sociobiology, 61(4), 517–522.



Research Article - Bees

Most read articles by the same author(s)

1 2 > >>