The honeybee (Apis mellifera L.), as a pollinating insect, plays an essential role in ecosystems. Approximately 90% of wild plants and a third of food plants consumed by humans depend on their pollination. It has been estimated that the economic value of bee pollination, for agriculture alone, could be around US $ 500 billion annually worldwide. This activity represents an important source of food and natural medicines for humans, without underestimating the commercial value of honey, wax, pollen, propolis and royal jelly (MITECO 2019MITECO (Ministerio para la Transición Ecológica y el Reto Demográfico, España). 2019. Borrador de plan de acción nacional para la conservación de los polinizadores. Versión 18 de enero de 2019, p. 73, Available: https://www.miteco.gob.es/es/biodiversidad/participacion-publica/borradorplanpolinizadores_tcm30-487605.pdf.).
In Cuba, the technology of pollination of crops with bees is hardly practiced, and the fundamental objective of beekeeping is to obtain the products of the hive, especially honey. Its importance as an export-generating activity has promoted cutting-edge technical advances worldwide, based on the professionalism of beekeepers, land use planning and integrated health management, which excludes the use of drugs and has developed, for years, a national genetic selection and improvement program (MINAG 2002MINAG (Ministerio de la Agricultura). 2002. Programa de mejoramiento genético de la abeja Apis mellifera Lcon abejas localmente adaptadas al ácaro Varroa destructor I. CIAPI, Cuba, p. 8., 2013MINAG (Ministerio de la Agricultura). 2013. Programa de mejoramiento genético de la abeja Apis mellifera con abejas localmente adaptadas al ácaro Varroa destructor II. CIAPI, Cuba, p. 22. and Pérez-Morfi and Pérez-Piñeiro 2018 Pérez-Morfi, A. & Pérez-Piñeiro, A. 2018. Experiencias sobre crianza y selección en la apicultura cubana. Memorias del XIII Congreso de la Federación Latinoamericana de Apicultura (FILAPI). 2-5 de agosto de 2018, Montevideo, Uruguay, p. 278.).
This progress has been made with a hybrid bee from European breeds, especially A. mellifera mellifera and A. mellifera ligustica, despite the presence of the dangerous mite Varroa destructor for 25 years. However, the possibilities of its improvement are not exhausted and the risk of introduction of the Africanized bee is permanent, and not only accidental, so it constitutes an environmental, social and economic threat that requires constant monitoring. Morphometric studies are quick and effective diagnostic methods to discard the presence of these bees, especially in Cuba, which is surrounded by territories where these highly defensive hybrids, with great expansion power, exist (Moreira et al. 2017Moreira, S.B.L.C., Sampaio, G., Alcântara, H., Alves, E., Santiago, D. & De Holanda J. 2017. “Infestação do ácaro Varroa destructor em colônias de abelhas africanizadas (Apis mellifera L.) no Semiárido potiguar, Nordeste do Brasil”. Revista Verde de Agroecologia e Desenvolvimento Sustentável, 12(1): 143-149, ISSN: 1981-8203, DOI: http://dx.doi.org/10.18378/rvads.v12i1.4845., Graciano 2018Graciano, L.A. 2018. Niveles de infestación de Varroa destructor (Mesostigmata: Varroidae) en abejas africanizadas (Apis mellifera scutellata hib). MSc. Thesis. Facultad de Ciencias, Universidad Nacional de Colombia, Medellín, Colombia, p. 87. and Urbina et al. 2019Urbina, R.A., Utrera-Quintana, F., Castillo-González, F., Livera-Muñoz, M., Benítez-Riquelme, I., Villa-Mancera, A.E., Hernández-Hernández, J.E. & Silva-Rojas, H.V. 2019. "Valoración del origen africanizado en la integración de una población experimental de Apis mellifera L". Revista Fitotecnia Mexicana, 42(2): 111-118, ISSN: 0187-7380.). Despite speculation about its presence on the island (Galindo-Cardona 2013Galindo-Cardona, A. 2013. "Especies Invasoras. Apis mellifera L., de África vinieron todas". Boletín de la Sociedad Entomológica Argentina, 23(1): 3-4, ISSN: 1666-4612.), there is no report that supports this assumption.
Drones provide the highest percentage of genotypic and phenotypic characteristics. They develop in cells larger than the workers, and are born 24 days after laying. For them, the queen bee breeding center is key, of which there are more than 60 in Cuba (APICUBA 2019APICUBA (Empresa Apícola Cubana). 2019. Informe Resumen Balance de Trabajo del Año 2019 y Objetivos 2020. Ministerio de la Agricultura, La Habana, Cuba, p. 17.). However, these drones have received little attention from the research point of view (Bande et al. 2008Bande, J.M., Verde, M. & Leal, A. 2008. Manual de Apicultura. Curso Consejo Científico Veterinario. La Habana, Cuba, p. 38. and Ocaña-Nápoles 2014Ocaña-Nápoles, L. 2014. Caracterización morfobiométrica de zánganos de Apis mellifera L. en un apiario de producción de la provincia de Mayabeque. Trabajo de Curso. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 27. ). European drones have a very bulky body and their weight can reach 230 mg, and between 15 and 17 mm in length (Pasini and Falda 2003Pasini, B. & Falda, M.T. 2003. L´allevamento d´apiregine. Una per tutte...tutte per una. Unione Nazionale Associazione Apicultori Italiani (U.N.A.API.), pp. 31-53. ). The Africanized ones are smaller (Guzmán-Novoa et al. 2011Guzmán-Novoa, E., Correa, B.A., Espinosa, M.L.G., Guzmán, N.G. 2011. "Colonización, impacto y control de las abejas melíferas africanizadas en México". Veterinaria México, 42(2): 149-178, ISSN: 0301-5092.), but faster. They are present throughout the year and produce almost 30% more sperm (Hernández 2019Hernández, C.U. 2019. Determinación de la africanización de las abejas melíferas (Apis mellifera L.) de la Comarca Lagunera. Diploma Thesis. Universidad Autónoma Agraria Antonio Narro, Coahuila, México, p. 51.).
It has been reported that among the most important environmental factors that can reduce size and weight of bees at birth is the decrease in the capacity of brood cells due to aging of honeycombs (Sanabria et al. 2015 Sanabria, J.L., Demedio, J., Pérez, T., Peñate, I., Rodríguez, D. & Lóriga, W. 2015. "Índices de infestación por Varroa destructor en colmenas sin medidas de control". Revista de Salud Animal, 37(2): 118-124, ISSN: 2224-4700.) and parasitization by Varroa destructor mite during the capping period (Guzmán-Novoa and Correa 2012Guzmán-Novoa, E. & Correa, B. 2012. Patología, diagnóstico y control de las principales enfermedades y plagas de las abejas melíferas. Ed. Imagen Editorial Yire. Ciudad de México, México, p. 165. and Ramsey 2018Ramsey, S. 2018. The acari varroa feeds mainly of the fatty corporal fabric of the melliferous bee. PhD Thesis. University of Maryland, Maryland, U.S.A., p. 143.). Although Cuba has maintained an efficient program for the integrated management of bee health for years, which excludes the use of chemical treatments (APICUBA 2019APICUBA (Empresa Apícola Cubana). 2019. Informe Resumen Balance de Trabajo del Año 2019 y Objetivos 2020. Ministerio de la Agricultura, La Habana, Cuba, p. 17.), varroosis continues to be the main problem worldwide (OIE 2015OIE (World Organisation for Animal Health). 2015. Infestación de las abejas melíferas por Varroa spp. (Varroosis). In: Código Sanitario para los Animales Terrestres. 15th Ed. Ed. Organización Mundial de Sanidad Animal. París, Francia, ISBN: 92-9044-679-X.). From this background, this research set out to determine biometric parameters of drones at birth and the factors that affect them.
The study was developed in three apiaries in Mayabeque province. One from San José de las Lajas municipality (SJ) and two from Tapaste town (T1 and T2), belonging to this municipality. SJ apiary is an auxiliary apiary of the Centro de Cría de Reinas “Los Pinos” (CCR, in Spanish). It has 20 hives with two bodies, in good condition, which supply honeycombs with pollen or brood when required. T1 and T2 apiaries have maintained a composition of 20 hives each, which objective is honey production.
Although these apiaries yielded a mean productivity of 40 kg/hive, the state of their brood chambers showed blackened honeycombs due to aging. These apiaries were chosen for not being sources of infectious diseases and for having access and protection facilities. The annual change of queens takes place in all hives.
From each apiary, 10 hives were analyzed, in two bodies and with a good strength state. As a requirement, hives had to have a brood of drones with pink eyes, about four or five days after hatching. In each hive, the diameter of the cells of drone was measured and a honeycomb fragment (10x10 cm) of this brood was cut for transfer to the laboratory and to conclude its development in an incubator.
A total of 50 drones per hive were evaluated at birth. A monthly sampling was carried out during November and December 2015 and January 2016. Body length (LZ) and forewing, weight (PZ), color and extension and invasion intensity (E.I. and I.I.) were determined to newly emerged drones for Varroa destructor. For this, the surface of individuals and the interior of the corresponding cells were examined in a stereoscopic microscope.
Cell diameter. One honeycomb per one of the 10 analyzed hives from the center of the brood chamber was selected. An amount of 10 cells were measured on both sides in line to average and reduce the error to result in four measurements per hive. Measurements were carried out with a special methodology of the program for controlling the Africanized bee (SARH-USDA 1986SARH-USDA. 1986. Regla para la medición del diámetro de las celdas de cría de obreras. Programa Conjunto México-EE.UU para el control de la abeja africanizada. Available: http://www.sat.gob.mx/sitio_internet/asistencia_contribuyente/información.).
Drone length. A mechanical Vernier caliper was used to determine body length of drones.
Right forewing length. A group of 25 drone wings were taken per hive. They were detached with the fingers and fixed with transparent commercial enamel on slides on each sheet, and measured with the aid of the caliper.
Weighing. The analyzed drones were numbed with ether and individually placed with the help of a tweezers on a Sartorius scale, BSA 124S model, on filter paper.
Coloration. It was carried out by visual appreciation and the criteria of Pérez Hernández (2009)Pérez Hernández, A. 2009. Índices de infestación por Varroa destructor y su relación con las características biométrico moleculares de Apis mellifera y el mecanismo defensivo ANF, en un CCR. MSc. Thesis. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 60. were considered, which establish the categories of drones: with totally yellow (A), totally black (N) and intermediate (I) segments. The latter are yellow in one or two segments.
Invasion extent and mean invasion intensity by V. destructor. A 10x10 cm honeycomb fragment was cut from each hive, with young in the pupal stage (16-17 d of age). They were brought to the laboratory in identified glass flasks, with perforated lids (NC 2010NC (Norma Cubana). 2010. Apicultura. Varroosis. Diagnóstico Veterinario. Oficina Nacional de Normalización, La Habana, Cuba.). They were placed in 30x20x10 cm cardboard boxes and placed in a total visibility incubator (Stuart Scientific Incubator SI60) at 34oC. At the beginning of the hatching process, cells were opened and the interior and drones were examined with the help of a tweezer and a simple magnifying glass with 4x magnification. These indexes were calculated following Verde et al. (2013)Verde, M., Demedio, J. & Gómez, T. 2013. Apicultura. Salud y Producción. Tomo II. Guía técnica para el apicultor. 1st Ed. ISBN: 978-959-7190-21-9, p. 208. using the following formula:
Data was processed by Statgraphics, version 5.1 (Statistical Graphics Corp 2000Statgraphics Plus. 2000. Version 5.1 [Windows], Available: http://wwwstatgraphics.com/statgraphics/statgraphics.nsf/pd/pdpricing.):
Descriptive statistics, for morphometric indicators and infestation indexes
Comparison of means for the weight of parasitized and non-parasitized drones
Multiple range test (Duncan 1955Duncan, D.B. 1955. "Multiple Range and Multiple F Tests". Biometrics, 11(1): 1-42, ISSN: 0006-341X, DOI: https://doi.org/10.2307/3001478.) to compare all biometric parameters among the three apiaries
Simple regression analysis (cell diameter - PZ; I.I. - PZ; I.I. - LZ).
To compare the percentage values, a comparison of proportions was performed (Compaprop).
Mean diameter of cells in SJ apiary was superior to that of the commercial apiaries (table 1). As in addition to geographical proximity, most of the queens of the latter come from the CCR, it is unlikely that this difference and that between the T1 and T2 apiaries is due to genetic factors. Rather, it can be attributed to the way bees are forced to build these cells. In the CCR, drone rearing is favored in parental hives with naturally manufactured drone combs. However, in commercial apiaries, this type of cell is built on sheets for workers of less stamping and less, in lower marginal areas of these combs.
* Each measuring (40) corresponded to four segments of 10 cells
Multiple range test (Duncan 1955Duncan, D.B. 1955. "Multiple Range and Multiple F Tests". Biometrics, 11(1): 1-42, ISSN: 0006-341X, DOI: https://doi.org/10.2307/3001478.): P ˂ 0.05
Different letters indicate significant differences
These measures could be taken as a baseline for comparison in the absence of antecedents. Although reduction and variability has been reported in the diameter of worker cells, when bees are induced to work on sheets that do not have their own stamping dimensions (Hall et al. 2015Hall, H.G., Zettel-Nalen, C. & Ellis, J.D. 2015. African Honey Bee: What You Need to Know. Electronic Data Information Source Publication ENY-114 (MG113), Institute of Food and Agricultural Sciences Extension, University of Florida, U.S.A., Available: https://edis.ifas.ufl.edu/mg113, [Consulted: July 4th, 2017]. and Sanabria et al. 2015 Sanabria, J.L., Demedio, J., Pérez, T., Peñate, I., Rodríguez, D. & Lóriga, W. 2015. "Índices de infestación por Varroa destructor en colmenas sin medidas de control". Revista de Salud Animal, 37(2): 118-124, ISSN: 2224-4700.), cell by cell measurement and knowing its origin is required, as well as more precise instruments. The external diameter has, in addition of an identifying value, the advantages of being a highly heritable genetic trait, like other morphological ones (Guzmán-Novoa 2012Guzmán-Novoa, E. 2012. Manual para la cría de abejas reinas. Programa Nacional de Apicultura del INIFAP, SAGARPA. p. 39, Available: https://www.buenastareas.com/ensayos/Manual-Para-La-Cria-De-Abejas/2175540.html.) and easy to measure. Meanwhile, the environmental influence on cell capacity, caused by aging and the accumulation of residues in each rearing cycle, has been routinely evaluated based on the management of the rearing chamber and the coloration of combs (Verde et al. 2013Verde, M., Demedio, J. & Gómez, T. 2013. Apicultura. Salud y Producción. Tomo II. Guía técnica para el apicultor. 1st Ed. ISBN: 978-959-7190-21-9, p. 208.).
Body mean lengths drones at birth (table 2) are between 15.40 ± 0.70 mm and 15.57 ± 0.66 mm, the highest value corresponding to samples obtained from the SJ apiary.
Multiple range test (Duncan 1955Duncan, D.B. 1955. "Multiple Range and Multiple F Tests". Biometrics, 11(1): 1-42, ISSN: 0006-341X, DOI: https://doi.org/10.2307/3001478.)
Different letters indicate significant differences (P ˂ 0.05)
There is little history of this indicator in drone rearing. Arias et al. (2006)Arias, M.C., Rinderer, T.E. & Sheppard, W.S. 2006"Further characterization of honey bees from the Iberian Peninsula by allozyme, morphometric and mtDNA haplotype analyses". Journal of Apicultural Research, 45(4): 188-196, ISSN: 0021-8839, DOI: https://doi.org/10.1080/00218839.2006.11101346. reported a range of 15-17 mm in hives in Spain, in which the values of the present study are included. Ocaña-Nápoles (2014)Ocaña-Nápoles, L. 2014. Caracterización morfobiométrica de zánganos de Apis mellifera L. en un apiario de producción de la provincia de Mayabeque. Trabajo de Curso. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 27. determined, in the same area, a minimum value of 14 mm in yellow and intermediate color flying drones, and a maximum of 18 mm, in intermediate and black ones. These results agree with the European origin of the Cuban native bee (Pérez-Morfi and Pérez-Piñeiro 2018 Pérez-Morfi, A. & Pérez-Piñeiro, A. 2018. Experiencias sobre crianza y selección en la apicultura cubana. Memorias del XIII Congreso de la Federación Latinoamericana de Apicultura (FILAPI). 2-5 de agosto de 2018, Montevideo, Uruguay, p. 278.) and show no clear difference between months. On the other hand, the effects overlap, so that samples for each month, to a large extent, must correspond to cells taken in the previous month, a fact that is often obviated when trying to establish cause-effect relationships.
The greater mean body lengths of drones in the SJ apiary in comparison with the T1 and T2 apiaries contribute to the greater dimensions of cells and a better zootechnical management in the former. T1 and T2 apiaries are the subject of poor zootechnical attention, appreciated in the darker and aged combs. This factor results in a reduction in the size of individuals raised in their cells. A similar effect was manifested in wings and has been recognized in the breeding of workers (Verde et al. 2013Verde, M., Demedio, J. & Gómez, T. 2013. Apicultura. Salud y Producción. Tomo II. Guía técnica para el apicultor. 1st Ed. ISBN: 978-959-7190-21-9, p. 208.). In this same SJ apiary, Pérez-Hernández (2009)Pérez Hernández, A. 2009. Índices de infestación por Varroa destructor y su relación con las características biométrico moleculares de Apis mellifera y el mecanismo defensivo ANF, en un CCR. MSc. Thesis. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 60. determined means of 5.37 ± 1.5 mm and 5.35 ± 1.1 mm in cells of workers, all in correspondence with values described for European bees. When analyzing the factors that influenced these results, these authors refer to honeycomb aging as the main environmental component that reduced the dimensions of the emerging individuals. However, a similar effect cannot be discarded, reported by Hall et al. (2015)Hall, H.G., Zettel-Nalen, C. & Ellis, J.D. 2015. African Honey Bee: What You Need to Know. Electronic Data Information Source Publication ENY-114 (MG113), Institute of Food and Agricultural Sciences Extension, University of Florida, U.S.A., Available: https://edis.ifas.ufl.edu/mg113, [Consulted: July 4th, 2017]. in cells infested by V. destructor.
In the same way that cell dimensions (Sanabria 2007Sanabria, J. L. 2007. Índices de infestación, estatus racial y expresión de mecanismos de resistencia en colmenas sin control antiVarroa. PhD Thesis. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 115.) are influenced by the fact that the bee must act on sheets with different patterns to their measurements, their cells do not correspond to the natural ones (Sanford and Hall 2005Sanford, M. & Hall, H. 2005. African Honey Bee: What You Need to Know. Available: http://edis.ifas.ufl.edu/BODY_MG113, [Consulted: March 13th, 2011].). In the SJ apiary (CCR), drone cells are found in naturally constructed combs, without having stamped sheets as a base. This helps to explain why they reach larger dimensions with respect to commercial apiaries (T1 and T2).
When comparing the results in the three months of study, differences were appreciated between December and January. However, both months did not differ from November (table 3), which is attributed to the age of the combs.
Multiple range test (Duncan 1955Duncan, D.B. 1955. "Multiple Range and Multiple F Tests". Biometrics, 11(1): 1-42, ISSN: 0006-341X, DOI: https://doi.org/10.2307/3001478.)
Different letters indicate significant differences P ˂ 0.05
Pérez Hernández (2014)Pérez Hernández, A. 2014. Índices de infestación por Varroa destructor y mecanismos de defensa, y su correspondencia con la supuesta condición de “europeas” y selectas de las colmenas en un Centro Genético de Producción de abejas reinas. PhD Thesis. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 101. observed that the forewing length of Cuban worker bees (9.55-9.70 mm) exceed those reported for American ecotypes of European ancestors (Uribe et al. 2003Uribe, R., Guzmán-Novoa, G., Hunt, G., Correa, B. & Zozaya, R. 2003. "Efecto de la africanización sobre la producción de miel, comportamiento defensivo y tamaño de las abejas melíferas (Apis mellifera L.) en el Altiplano Mexicano". Veterinaria México, 34(1): 47-59, ISSN: 0301-5092 ), which confirms the first and most important observations made in Cuba (Díaz-Millán 1981Díaz-Millán, M.E. 1981. "Características morfológicas de la abeja (Apis mellifera) en las diferentes regiones de Cuba, donde se seleccionarán colmenas para constituir apiarios de reserva". Agrotecnia de Cuba, 13(1): 8-17, ISSN: 2414-4673, 0568-3114.). Similar performance is to be expected in drones, clearly larger than Africanized ones. Despite the high heritability of this trait (Guzmán-Novoa 2012Guzmán-Novoa, E. 2012. Manual para la cría de abejas reinas. Programa Nacional de Apicultura del INIFAP, SAGARPA. p. 39, Available: https://www.buenastareas.com/ensayos/Manual-Para-La-Cria-De-Abejas/2175540.html.), possible environmental influences are not excluded, especially the aging of the brood chamber and the reduction of body and its appendages (Verde et al. 2013Verde, M., Demedio, J. & Gómez, T. 2013. Apicultura. Salud y Producción. Tomo II. Guía técnica para el apicultor. 1st Ed. ISBN: 978-959-7190-21-9, p. 208. and Hall et al. 2015Hall, H.G., Zettel-Nalen, C. & Ellis, J.D. 2015. African Honey Bee: What You Need to Know. Electronic Data Information Source Publication ENY-114 (MG113), Institute of Food and Agricultural Sciences Extension, University of Florida, U.S.A., Available: https://edis.ifas.ufl.edu/mg113, [Consulted: July 4th, 2017]. ).
Table 4 shows the mean length of the right forewing between 11.31 ± 0.46 mm and 11.37 ± 0.48 mm, without a significant superiority, in the SJ apiary.
Multiple range test (Duncan 1955Duncan, D.B. 1955. "Multiple Range and Multiple F Tests". Biometrics, 11(1): 1-42, ISSN: 0006-341X, DOI: https://doi.org/10.2307/3001478.): P ˂ 0.05 (n.s.)
In Cuba, there is no history of published studies, in which wing dimensions of drones are determined. Díaz-Millán (1981)Díaz-Millán, M.E. 1981. "Características morfológicas de la abeja (Apis mellifera) en las diferentes regiones de Cuba, donde se seleccionarán colmenas para constituir apiarios de reserva". Agrotecnia de Cuba, 13(1): 8-17, ISSN: 2414-4673, 0568-3114. carried out the most extensive and documented morphometric evaluations in Cuba, but all were conducted in worker bees. Studies of Voroshilov (2008)Voroshilov, U. 2008. Determinación del estatus racial y parasitológico (varroosis) de colmenas (Apis mellifera L.), ante la amenaza de la africanización. Diploma Thesis. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 71. and Pérez Hernández (2014)Pérez Hernández, A. 2014. Índices de infestación por Varroa destructor y mecanismos de defensa, y su correspondencia con la supuesta condición de “europeas” y selectas de las colmenas en un Centro Genético de Producción de abejas reinas. PhD Thesis. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 101. did not considered the males either. It is significant that, in all these studies, the wings of workers reached dimensions in the range of those of Europe (Arias et al. 2006Arias, M.C., Rinderer, T.E. & Sheppard, W.S. 2006"Further characterization of honey bees from the Iberian Peninsula by allozyme, morphometric and mtDNA haplotype analyses". Journal of Apicultural Research, 45(4): 188-196, ISSN: 0021-8839, DOI: https://doi.org/10.1080/00218839.2006.11101346.) or exceeded them. This result contributes to refute the speculation about the presence of the Africanized bee in Cuba (Galindo-Cardona 2013Galindo-Cardona, A. 2013. "Especies Invasoras. Apis mellifera L., de África vinieron todas". Boletín de la Sociedad Entomológica Argentina, 23(1): 3-4, ISSN: 1666-4612.).
The mean weight of drones at birth (table 5) is between 190.67 mg and 200.13 mg, with the lowest values in the T1 apiary and the tendency to supremacy in SJ.
Multiple range test (Duncan 1955Duncan, D.B. 1955. "Multiple Range and Multiple F Tests". Biometrics, 11(1): 1-42, ISSN: 0006-341X, DOI: https://doi.org/10.2307/3001478.)
Different letters indicate significant differences P ˂ 0.05
It is recognized that, as the size of the brood comb cell increases, an increase of the weight of the emerging drone is obtained under the given conditions. A positive correlation (P ˂ 0.10), but weak, was observed between the diameter of the brood cells and the weight of drones at birth (table 6), mainly attributed to the mentioned environmental effects. The greatest weight differences were observed between the SJ and T1 apiaries, but it was not possible to determine the number of cycles developed in the sampled combs. In Spain, Arias et al. (2006)Arias, M.C., Rinderer, T.E. & Sheppard, W.S. 2006"Further characterization of honey bees from the Iberian Peninsula by allozyme, morphometric and mtDNA haplotype analyses". Journal of Apicultural Research, 45(4): 188-196, ISSN: 0021-8839, DOI: https://doi.org/10.1080/00218839.2006.11101346. reported a mean similar to November (200 mg), also in drones at birth. Meanwhile, Bande et al. (2008)Bande, J.M., Verde, M. & Leal, A. 2008. Manual de Apicultura. Curso Consejo Científico Veterinario. La Habana, Cuba, p. 38., in Cuba, obtained a mean of 280.12 mg, but from already fed individuals. This result is superior to that indicated by Ocaña-Nápoles (2014)Ocaña-Nápoles, L. 2014. Caracterización morfobiométrica de zánganos de Apis mellifera L. en un apiario de producción de la provincia de Mayabeque. Trabajo de Curso. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 27. , also for flying drones (233 mg) in the area of San José de las Lajas.
Although in appearance, this result would be in contradiction with the previous ones, it should be considered that it includes all cells, where different effects overlap, and some are opposite. Since it is not possible to determine the internal diameters, larger cells may be older and, therefore, have their capacity decreased. In addition, the intensity of parasitization by Varroa also interacts and results in a distorting effect.
There does not seem to be a clear explanation for the differences in mean weight of drones between November and December-January (table 7).
Multiple range test (Duncan 1955Duncan, D.B. 1955. "Multiple Range and Multiple F Tests". Biometrics, 11(1): 1-42, ISSN: 0006-341X, DOI: https://doi.org/10.2307/3001478.)
Different letters indicate significant differences P ˂ 0.05
It is probable that this difference expresses the result of interactions among several factors, such as the quantity and quality of food. In turn, they depend, for their production, on the reserves of bee bread, the entry of freshly harvested pollen and the availability of honey or nectar. At the same time, the production of these foods is conditioned by the numbers of foraging and nurse bees. As all these factors are highly variable and their status may reflect prior deficiencies or imbalances, it is extremely difficult to determine the causes in a short-term, specific study. To this should be added the fact that the production of drones does not follow the same patterns as that of workers and is more dependent on the abundance of food and the time of year (Verde et al. 2012Verde, M., Gómez, T. ( Demedio J. 2012. Salud Apícola. Tomo I. Generalidades. 1st Ed. ISBN: 978-959-7190-15-8, p. 207.).
All drones examined at birth (3 071) were black. It is surprising that Ocaña-Nápoles (2014)Ocaña-Nápoles, L. 2014. Caracterización morfobiométrica de zánganos de Apis mellifera L. en un apiario de producción de la provincia de Mayabeque. Trabajo de Curso. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 27. has determined, in the same area, 60% intermediate color, 18% black and 22% yellow, but in individuals captured at random within the hive. In the absence of paternal inheritance in drones, the explanation should be in the change of queens, which is carried out at least once a year, although it is necessary to deepen the matter.
The extent of invasion in this brood was high, to the extent that its mean values exceeded 70%, in SJ as in the hives of T1 and T2 (table 8). These values are at the level of those determined in 1996-1997 by Demedio (2001)Demedio, J.L. 2001. La varroosis de las abejas en una zona de la provincia de La Habana. Agente etiológico, índices de infestación y control biotécnico y químico. PhD Thesis. Facultad de Medicina Veterinaria, Universidad Agraria de La Habana, San José de las Lajas, Mayabeque. at the beginning of the epizooty of varroosis, in San José de las Lajas municipality. They are also consistent with reports by Sanabria (2007)Sanabria, J. L. 2007. Índices de infestación, estatus racial y expresión de mecanismos de resistencia en colmenas sin control antiVarroa. PhD Thesis. Universidad Agraria de La Habana, San José de las Lajas, Mayabeque, Cuba, p. 115. and Sanabria et al. (2015) Sanabria, J.L., Demedio, J., Pérez, T., Peñate, I., Rodríguez, D. & Lóriga, W. 2015. "Índices de infestación por Varroa destructor en colmenas sin medidas de control". Revista de Salud Animal, 37(2): 118-124, ISSN: 2224-4700., also in the same area. Apparently, the situation is unusual because then, the first contact of the local bee with the parasite occurred. Meanwhile, by 2016, there was a history of coexistence. According to some criteria (Pérez-Morfi and Pérez-Piñeiro 2018 Pérez-Morfi, A. & Pérez-Piñeiro, A. 2018. Experiencias sobre crianza y selección en la apicultura cubana. Memorias del XIII Congreso de la Federación Latinoamericana de Apicultura (FILAPI). 2-5 de agosto de 2018, Montevideo, Uruguay, p. 278.), the current bees are descendants of those that survived the initial impact of the mite, a result of natural selection, and the subsequent work of the selection and improvement program that continues today. Apparently, it is not appropriate to talk about natural selection in intensive beekeeping.
Comparison of proportions: F= 1.01 n.s.
Multiple range test (Duncan 1955Duncan, D.B. 1955. "Multiple Range and Multiple F Tests". Biometrics, 11(1): 1-42, ISSN: 0006-341X, DOI: https://doi.org/10.2307/3001478.): P ˂ 0.05
Different letters indicate significant differences
The current high values could be explained by the scarcity of this type of breeding in the evaluated months, which, together with its greater and longer attractiveness for the founding females (Granadillo 2009Granadillo, N. 2009. Índices de infestación y reproducción del ácaro Varroa destructor en colmenas de abejas africanizadas. Diploma Thesis. Venezuela.), results in a high percentage of parasitized cells. Drijfhout et al. (2005)Drijfhout, K., Kochansky, J., Lin, F. & Calderone, N.W. 2005. " Components of Honeybee Royal Jelly as deterrents of the Parasitic Varroa Mite, Varroa destructor". Journal of Chemical Ecology, 31(8): 1747-1764, ISSN: 1573-1561, DOI: https://doi.org/10.1007/s10886-005-5925-6. observed that varroa mites show a preference for drone brood up to nine times greater, and that royal jelly exerts a deterrent effect on queen cells. Especially for CCR, these situations should call attention to the need to apply an organic treatment strategy that, due to its safety and efficacy (Reyes 2016Reyes, F. 2016. Efectividad de cuatro acaricidas en el control del ácaro (Varroa destructor) en abejas (Apis mellifera L.). MSc. Thesis. Universidad Nacional Agraria, Lima, Perú, p. 88. ), allows these indexes to be reduced.
In the search for a possible effect of the parasite on the morphological parameters of drones at birth, invasion intensity is the quantitative indicator that allows determining a possible influence. However, the combination of both offers an evaluative criterion of the severity of the parasitic manifestation, as it is a cloistered host that is affected, at this stage, only by the founding females and their offspring.
As can be observed, invasion intensity was lower in the SJ apiary, although it should be considered that the mean difference does not exceed unity, that is, an individual. Something similar can be seen in the analysis per months (table 9). Despite a specific drop that is difficult to explain, in December (one sampling), November and January, the same values and mean invasion intensity are presented, although with statistical differences. Due to the effect, they should not significantly differ, as they do not even reach half a unit.
Comparison of proportions: F= 6.55**
Multiple range test (Duncan 1955Duncan, D.B. 1955. "Multiple Range and Multiple F Tests". Biometrics, 11(1): 1-42, ISSN: 0006-341X, DOI: https://doi.org/10.2307/3001478.): P ˂ 0.05
Different letters in the same line indicate significant differences
By taking invasion intensity as an independent variable and the morphometric parameters of drones at hatching (body length and weight) as dependent variables, a weak or non-significant relationship was determined (tables 10 and 11).
It is evident that the relatively low general mean load of parasites per cell did not significantly influence mean weight and length of newborn individuals. However, Getchev (1995)Getchev, I. 1995. "Effects of parasitism by Varroa jacobsoni on morphometrics of Apis mellifera worker honey bees”. Veterinarski Archiv, 65(6): 193-197. observed a weight loss between 11 and 19% according to the infestation rate, which causes a decrease in their flight capacity. This situation depends on the number of mother mites, although Rosenkranz et al. (2010)Rosenkranz, P., Aumeier, P. & Ziegelmann, B. 2010. "Biology and control of Varroa destructor”. Journal of Invertebrate Pathology, 103: S96-S119, ISSN: 0022-2011, DOI: https://doi.org/10.1016/j.jip.2009.07.016. stated that a single female of V. destructor can produce a mean loss of 7% of the bee weight. Although mites cause a significant reduction of weight and size of workers, a similar loss was not observed in the results with the drones of the current study. Likewise, it was manifested when comparing mean values of this parameter (table 12) with the inclusion of all the parasitized cells.
Comparison of means: N= 180; p = 0.41. (n.s.)
The result was different when comparing the weights of hatched individuals from cells with 5-6 mites, but not with 3-4 mites and those not parasitized (tables 13 and 14).
Comparison of means: N= 122; p= 0.54 n.s.
Comparison of means: N= 38; p= 0.005
It was confirmed that, to produce a significant effect of reduction in the morphometric parameters under the conditions of this study, there must be an invasion intensity superior to 4. However, it is likely that the concurrence of other factors of a nutritional or biological nature may promote measurable quantitative effects, with lower infestation rates.
An already known effect of reducing the weight of worker bees when they hatch is visible, which is the result, among other factors, of the spoilage action of adult females and juvenile stages of the mite. In this case, it is evident in drones, which breeding is more attractive and leads to greater reproductive success (Granadillo 2009Granadillo, N. 2009. Índices de infestación y reproducción del ácaro Varroa destructor en colmenas de abejas africanizadas. Diploma Thesis. Venezuela.), in addition to a proven harmful action on the immune system (Ramsey 2018Ramsey, S. 2018. The acari varroa feeds mainly of the fatty corporal fabric of the melliferous bee. PhD Thesis. University of Maryland, Maryland, U.S.A., p. 143. and Larsen et al. 2019Larsen, A., Reynaldi, F.J. & Guzmán-Novoa, E. 2019. "Bases del sistema inmune de la abeja melífera (Apis mellifera). Revisión Bibliográfica". Revista Mexicana de Ciencias Pecuarias, 10(3): 705-728, ISSN: 2448-6698, DOI: https://doi.org/10.22319/rmcp.v10i3.4785.). This fact is of special interest in the paternal hives from the CCR, and this is because, besides drone breeding is favored, the application of varroicidal treatments is prohibited, even those accepted by the requirements of organic beekeeping (MINAG 2013MINAG (Ministerio de la Agricultura). 2013. Programa de mejoramiento genético de la abeja Apis mellifera con abejas localmente adaptadas al ácaro Varroa destructor II. CIAPI, Cuba, p. 22.).
Morphological determinations indicate that they are drones of European lineage and a negative influence of parasitism by V. destructor mite was verified. High infestation rates were found in commercial hives, and most alarming, in paternal hives of a CCR. They were favored by their naturally abundant presence and the non-application of organic treatment, despite being accepted by the rigorous requirements established by the beekeeping production of this category.
The studied area lacks studies that are in correspondence with the value that must be restored to male bees. These results are just one step on the path to biometric characterization of drones and the factors that affect them. A larger-scale study is required, in which drones of the country are characterized, including physiological parameters such as sperm counts and their relationship with the quality of the queens produced in multiplying centers, without omitting their subsequent performance in the production hives.