Open access journal: http://periodicos.uefs.br/index.php/sociobiology ISSN: 0361-6525 DOI: 10.13102/sociobiology.v69i3.7729Sociobiology 69(3): e7729 (September, 2022) Introduction The Spanish word ganado (livestock) is directly derived from the verb ganar (to gain) and has several definitions, available in the tercentenary edition (the most updated one) of the Spanish language dictionary. Among these definitions, livestock is: “a group of beasts that feed and move together”, such as sheep, goats, and cattle. In its second definition, current since 1803, livestock is also defined as the group of bees in a beehive (RAE, 2017). Abstract The knowledge of the different plant species that make up the feeding diet of animals is highly important to develop more efficient strategies. This research aimed to characterize the food potential available for the Cuban stingless bee livestock of the Matanzas and Mayabeque provinces. Palynological analysis was done using 60 g of pollen from sealed pots and 80 mL of honey from the ten randomly selected beehives (five in each province). The results showed that in the honey collected in Matanzas province, the most represented family was Amaranthaceae, followed by Myrtaceae and Fabaceae. Meanwhile, for Mayabeque, the most represented ones were the families Fabaceae and Myrtaceae. Regarding the stingless bee pollen of Matanzas provenance, the family Fabaceae prevailed, followed by Burseraceae and Myrtaceae. The pollen corresponding to Mayabeque coincided in showing Fabaceae as the most representative. In addition, pollen grains of small size (from 10 to 25 µm) were collected, with a marked representation of the pollen type of Mimosa pudica in the Mayabeque honey. It was concluded that the Cuban stingless bee livestock of the Matanzas and Mayabeque provinces had a specialist feeding behavior because a low number of plant taxa made up its diet. Sociobiology An international journal on social insects Leydi F Carballo1, David R de la Cruz2,4, José S. Sanchez2,4, Estefanía S. Reyes2, Walberto L Peña3 Jorge D Lorenzo3, Maykelis D Solares5, Dariel M Querol1 Article History Edited by Evandro Nascimento Silva, UEFS, Brazil Received 12 January 2022 Initial acceptance 04 March 2022 Final acceptance 16 July 2022 Publication date 07 September 2022 Keywords Honey, stingless bee pollen, Melipona beecheii, melissopalynology. Corresponding author Leydi Fonte Carballo Estación Experimental de Pastos y Forrajes “Indio Hatuey” Central España Republicana, Matanzas, Cuba. E-Mail: leydis.fonte@ihatuey.cu Taking the previous concept as a reference, Lóriga (2015) defines the Cuban stingless bee livestock as all the managed beehives of the bee species Melipona beecheii Bennett, 1831 (Apidae: Meliponini), included in a census or not. According to Sáenz et al. (2000), during the foraging process, bees visit flowers in search of food, then pollen grains adhere to their body and later become part of honey. The palynology branch studying these grains to know the botanical origin of the different products elaborated or stored in the hives of diverse groups of bees is called melissopalynology. 1 - Estación Experimental de Pastos y Forrajes “Indio Hatuey” Central España Republicana, Matanzas, Cuba 2 - Grupo de Palinología y Conservación Vegetal (GPCV), Instituto Hispano Luso de Investigaciones Agrarias (CIALE), Universidad de Salamanca, España 3 - Universidad Agraria de La Habana, Facultad de Medicina Veterinaria. Carretera Tapaste y Autopista Nacional, San José de las Lajas, Mayabeque, Cuba 4 - Departamento de Botánica y Fisiología Vegetal, Universidad de Salamanca, España 5 - Independent researcher, USA RESEARCH ARTICLE - BEES Cuban stingless bee livestock exhibit specialized floral resource use: a palynological study on honey samples from Matanzas and Mayabeque provinces mailto:leydis.fonte@ihatuey.cu Fonte-Carballo et al. – Cuban stingless bees with the specialized resource use2 With the use of this discipline, the study of the floral sources visited by bees is facilitated to a large extent (Sánchez, 2001). The antecedents of using trophic resources by bees allow the evaluation of the plant taxa that contribute nectar and/or pollen to the bee diet. And consequently, from these data, the elaboration of floristic lists and phenological calendars can be done, contributing to better management of the beehives by farmers (Flores et al., 2015). Due to the above-exposed facts, knowing the main plant species that make up the diet of these animals is a priority in developing efficient feeding strategies that guarantee higher production with an optimum health level. This research aimed to characterize the food potential available for the Cuban stingless bee livestock of the Matanzas and Mayabeque provinces and its possible application to other territories with beehives of this species. Materials and Methods Study sites The study was conducted in two Cuban stingless bee apiaries in April 2018. The first one is located in the Matanzas municipality, of Matanzas province, at 23°02’25’’ North latitude and 81°30’58’’ West longitude, at an altitude of 14 m.a.s.l.; and the second one, in the San Nicolás municipality, belonging to the Mayabeque province, located at 22°47’13’’ North latitude and 80°55’05’’ West longitude, at an altitude of 34 m.a.s.l. The characteristic climate of the studied area is classified as tropical, with marked seasonality of the rains in two periods corresponding to the dry season (DS), which extends from November to April, and the rainy season (RS), which lasts from May to October (Academia de Ciencias de Cuba, 1989). During the sampling period, the Matanzas locality showed a warm climate that caused minimum temperature values and positive anomalies of 0.8ºC. The recorded cumulative rainfall was 22.8 mm, lower than the historical mean (52.6 mm). Regarding relative humidity, it was 74%, over the historical values. In the case of Mayabeque, it was also warm, conditioned by the influence of high oceanic pressure that weakly influenced the territory during most of the month. This situation is responsible for the cold air masses not reaching the territory, which provoked temperature values with significant positive anomalies of 1.7 ºC compared with the historical average. The minimum temperatures averaged 19.9ºC. Regarding rainfall, the cumulative exceeded the historical mean of 171.3 mm, and the mean air relative humidity was 75%, slightly higher than the historical average. Floristic inventory of each stingless bee apiary Knowing the foraging area of bees is important to understand the use of floristic resources. Studies on species of the genus Melipona Illiger‎, 1806‎, found flight radios close to 2 km, with the studied species having body sizes larger than M. beecheii (Nunes Silva et al., 2020). However, a flight radius of 1 km was estimated for M. beecheii, similar to that of species of the genus Trigona Jurine, 1807 (Araújo et al., 2004), since they share similar biometric parameters (Fonte, 2007). Considering that both stingless bee apiaries were located in urban areas, where gardens and house backyards prevail, the main existing plant species potentially contributing with pollen and nectar were determined: native, naturalized, and ornamental plants (Pérez-Piñeiro, 2017). In addition, to determine the representativeness of the species in the environment of stingless bee apiaries, four categories (abundant, frequent, little frequent, and rare or scarce) and their corresponding biological form (tree, shrub, and herbaceous) were established, adapting the methodology proposed by García- Abad (2015) to urban environments, which are less diverse from the botanical point of view (Wittig & Becker, 2010). Construction of the palynotheque From each inventoried species, floral buds were taken and preserved in a universal fixator preservative composed of a mixture of 95º ethylic alcohol, glacial acetic acid, 37- 40% formaldehyde, and distilled water. Afterward, they were dissected with the aid of a surgical blade, extracting the anthers, which were placed in conical test tubes of 10 ml capacity, and the protocol used for the preparation of the samples for the palynological and melissopalynological qualitative analysis was followed. The photographic shot of each grain from the previously identified botanical species of reference was done through a camera (LEICA DFC290) coupled to the optical microscope (NIKON OPTIPHOT-I) with a magnification of 400. The Leica Application Suite, version 3.3.1 for Windows XP, was used for this purpose. The obtained photographs and bibliographic references (Aira et al., 2018, Pérez, 2016) served as a basis for determining pollen types found in the honey and pollen samples. Sample collection From the ten beehives under study in each province, five were randomly selected in each locality. In Mayabeque, beehives 14, 2, 48, 5, and 6 were chosen, while in Matanzas, beehives 1, 9, 11, 12 and 13 were selected. Sixty grams of stingless bee pollen from sealed pots and 80 mL of honey in each one of them were taken. Sampling was done in April 2018. Melissopalynological qualitative analysis The protocol used for sample preparation was that of honey analysis proposed by the Honey Work Group belonging to the Spanish Society of Palynology (APLE, for its initials in Spanish), recently constituted (Sánchez et al., 2018, González- Porto et al., 2018). The protocol is based, with modifications, on García et al. (2001), consisting of the previous treatment Sociobiology 69(3): e7729 (September, 2022) 3 of the samples with 5% acidulated water with their respective washings and centrifugations to obtain sediment that is in turn deposited on the microscope slide and one drop of glycerin jelly with fuchsin is applied before taking it to the microscope for its later analysis. Palynological qualitative analysis of stingless bee pollen One gram of sample was weighed and was subject to the above-explained protocol, with modifications in the volume of 5% acidulated water that was added. This methodology was used to liken it to the one used in the processing of honey samples, also considering the collection conditions of stingless bee pollen, with different load sizes, variable from small masses of 5-6 mm to dust particles. The reading of stingless bee honey and pollen samples was done under the optical microscope with a magnification of 400, identifying a minimum number of 300 to 500 pollen grains to guarantee correct botanical and geographical characterization. The samples were done in quadruple. The observed pollen types were placed in the different class frequencies according to Loveaux et al. (1978), as shown below:  Dominant pollen (>45%)  Companion pollen (16-45%)  Important isolated pollen (4-15%)  Rare or sporadic pollen (1-3%)  Present pollen (<1%) The Shannon-Weaver diversity index was used to determine the browsing strategy deployed by M. beecheii from the pollen diversity found in the honey samples (Shannon & Weaver, 1949). Where: H’= Shannon-Wiener Index. pi = Proportion of each pollen type found. ln = Natural logarithm. Meanwhile, the browsing uniformity of the bees was determined through Pielou’s evenness Index (Pielou, 1984). Where: J’= Browsing uniformity of the bees. H’= Shannon-Wiener index. H’ max = Napierian logarithm of pollen richness. The diversity (H’) and evenness (J’) indices were determined through the utilization of the software Diversity Species & Richness 3.02 (Henderson & Seaby, 2002). Results The results of the floristic inventory in both localities show that in the flight environment of stingless bees, there are 16 plant species in common. We identified 42 plants belonging to 27 families in Matanzas (Table 1). Alternanthera sessilis (L.) R. Br. ex DC, Mangifera indica L., Adonidia merrillii (Becc.) Becc, Bidens pilosa L., Bursera simaruba (L.) Sarg, Senna spectabilis (DC.) H.S. Irwin & Barneby, Persea americana Mill., Musa x paradisiaca L., Psidium guajava L. and Coccoloba uvifera (L.) L. were the most abundant. Table 1. Floristic inventory of the Matanzas locality, Cuba, 2018. Matanzas meliponary Family Scientific name Biological form Appearance frequency Amaranthaceae Alternanthera sessilis (L.) R. Br. ex DC. Herbaceous Abundant Anacardiaceae Mangifera indica L. Tree Abundant Anacardiaceae Anacardium occidentale L. Tree Rare or Scarce Annonaceae Annona muricata L. Tree Frequent Annonaceae Annona squamosa L. Tree Little Frequent Arecaceae Cocos nucifera L. Tree Frequent Arecaceae Roystonea regia (Kunth) O.F. Cook Tree Little Frequent Arecaceae Adonidia merrillii (Becc.) Becc. Tree Abundant Asteraceae Bidens pilosa L. Shrub Abundant Asteraceae Viguiera dentata Spreng. Shrub Little Frequent Bignoniaceae Crescentia cujete L. Tree Little Frequent Boraginaceae Cordia collococca L. Tree Rare or Scarce Boraginaceae Bourreria virgata (Sw.) G. Don Shrub Frequent Boraginaceae Cordia alliodora (Ruiz & Pav.) Oken Tree Frequent Burseraceae Bursera simaruba (L.) Sarg Tree Abundant Cactaceae Nopalea auberi (Pfeiff.) Salm-Dyck Tree Rare or Scarce Fonte-Carballo et al. – Cuban stingless bees with the specialized resource use4 Calophyllaceae Calophyllum antillanum Britton Tree Little Frequent Combretaceae Terminalia catappa L. Tree Frequent Commelinaceae Tradescantia pallida (Rose) D.R. Hunt Herbaceous Frequent Convolvulaceae Ipomoea triloba L. Herbaceous Frequent Convolvulaceae Turbina corymbosa (L.) Raf. Herbaceous Frequent Cucurbitaceae Cucurbita pepo L. Herbaceous Little Frequent Fabaceae Mimosa pudica L. Herbaceous Little Frequent Fabaceae Vachellia farnesiana (L.) Wight & Arn. Tree Little Frequent Fabaceae Senna spectabilis (DC.) H.S. Irwin & Barneby Tree Abundant Fabaceae Tamarindus indica L. Tree Rare or Scarce Fabaceae Trifolium incarnatum L. Herbaceous Frequent Fabaceae Dichrostachys cinerea (L.) Wight & Arn. Tree Frequent Lamiaceae Tectona grandis L. f. Tree Rare or Scarce Lauraceae Persea americana Mill. Tree Abundant Malvaceae Hibiscus rosa-sinensis L. Shrub Frequent Malvaceae Hibiscus elatus Sw. Tree Rare or Scarce Meliaceae Swietenia mahagoni (L.) Jacq. Tree Rare or Scarce Moraceae Morus alba L. Tree Rare or Scarce Moringaceae Moringa oleifera Lam. Tree Frequent Musaceae Musa x paradisiaca L. Herbaceous Abundant Myrtaceae Psidium guajava L. Tree Abundant Passifloraceae Passiflora edulis Sims Herbaceous Little Frequent Polygonaceae Coccoloba uvifera (L.) L. Tree Abundant Rutaceae Citrus x aurantium L. Tree Frequent Rutaceae Citrus x limon (L.) Osbeck Tree Frequent Sapotaceae Manilkara zapota (L.) P. Royen Tree Frequent Table 1. Floristic inventory of the Matanzas locality, Cuba, 2018. (Continuation) Matanzas meliponary Family Scientific name Biological form Appearance frequency In Mayabeque (Table 2), a lower number of plant species (33) was identified, grouped in 22 families. In the category of abundant are Mangifera indica L., Bursera simaruba (L.) Sarg, Ipomoea triloba L., Vachellia farnesiana (L.) Wight & Arn., Gliricidia sepium (Jacq.) Kunth, Persea americana Mill., Psidium guajava L., Gouania polygama (Jacq.) Urb. In general, in the two localities, the biological form that prevailed was the tree, followed by shrubs and herbaceous. In the honey samples from Matanzas (Table 3) higher richness of pollen forms is observed, with a mean of 11.2 and a maximum of 12 (beehives 9 and 11). In the analyze honey from this locality, the predominant pollen percentage was that of Alternanthera Forssk., with a mean of 38.26%, with the dominant pollen (>45%) in three of the five samples, followed by Psidium guajava L. (17.50%) and the genus Trifolium L. (16.07%). The most represented family was Amaranthaceae, followed by Myrtaceae, and finally Fabaceae because only one sample (beehive 9) showed pollen of Mimosa pudica L., justifying its inclusion in the companion category, being lower than in the other beehives. Meanwhile, in the samples of the Mayabeque provenance (Table 4), the quantity of pollen forms or types is considerably lower, with a mean value of 7 and a maximum of 13 for the case of the honey belonging to beehive 6. It should be highlighted that the pollen from Mimosa pudica prevailed in all the examined honey samples, with a mean percentage of 74.05%, besides being the dominant pollen (>45%) in the five studied beehives. Fabaceae and Myrtaceae, represented by the species Psidium guajava, stand out as companion pollen (16-45%) of the honey obtained from beehives 2 and 48, with a mean of 23.21%. When analyzing the foraging strategy of M. beecheii (Fig 1), we found that in the Matanzas honey, the mean value of the diversity index (H´) was 1.34, with a maximum value in beehive 11 (1.88). On the contrary, Mayabeque showed a lower mean (0,84). These results are in correspondence with the ones exposed in Table 4. Regarding the evenness index (Fig 1), in Matanzas, the mean value was 0.45, and again in beehive 11, the highest value was recorded (0.63). For Mayabeque, this indicator had a mean value of 0.28. Sociobiology 69(3): e7729 (September, 2022) 5 Mayabeque meliponary Family Scientific name Biological form Appearance frequency Anacardiaceae Mangifera indica L. Tree Abundant Arecaceae Cocos nucifera L. Tree Frequent Arecaceae Roystonea regia (Kunth) O. F. Cook Tree Frequent Arecaceae Adonidia merrillii (Becc.) Becc. Tree Frequent Asteraceae Bidens pilosa L. Shrub Frequent Boraginaceae Cordia gerascanthus L. Tree Little Frequent Burseraceae Bursera simaruba (L.) Sarg Tree Abundant Cactaceae Dendrocereus nudiflorus (Engelm. ex Sauvalle) Britton & Rose Tree Little Frequent Convolvulaceae Ipomoea triloba L. Herbaceous Abundant Convolvulaceae Turbina corymbosa (L.) Raf. Herbaceous Frequent Ericaceae Lyonia myrtilloides Griseb. Tree Little Frequent Fabaceae Mimosa pudica L. Herbaceous Frequent Fabaceae Vachellia farnesiana (L.) Wight & Arn. Tree Abundant Fabaceae Gliricidia sepium (Jacq.) Kunth Tree Abundant Lauraceae Persea americana Mill. Tree Abundant Malvaceae Ceiba pentandra (L.) Gaertn. Tree Little Frequent Meliaceae Trichilia hirta L. Tree Little Frequent Moringaceae Moringa oleifera Lam. Tree Little Frequent Myrtaceae Psidium guajava L. Tree Abundant Nyctaginaceae Pisonia aculeata L. Shrub Frequent Oxalidaceae Averrhoa carambola L. Tree Rare or Scarce Polygonaceae Antigonon leptopus Hook. & Arn. Herbaceous Frequent Rhamnaceae Gouania polygama (Jacq.) Urb. Shrub Abundant Rosaceae Prunus domestica L. Tree Frequent Rosaceae Pyrus communis L. Tree Little Frequent Rubiaceae Coffea arabica L. Shrub Little Frequent Rutaceae Citrus x aurantium L. Tree Frequent Rutaceae Citrus x limon (L.) Osbeck Tree Frequent Sapindaceae Melicoccus bijugatus Jacq. Tree Frequent Sapindaceae Nephelium lappaceum L. Tree Rare or Scarce Sapindaceae Serjania diversifolia (Jacq.) Radlk. Herbaceous Little Frequent Sapotaceae Manilkara zapota (L.) P. Royen Tree Frequent Sapotaceae Pouteria dictyoneura (Griseb.) Radlk. Tree Little Frequent Table 2. Floristic inventory of the Mayabeque locality, Cuba, 2018. Fig 1. Values obtained in the diversity (H´) and evenness (J´) indices of the Matanzas (1, 9, 11, 12, 13) and Mayabeque honeys (2, 14, 48, 5, 6). Fonte-Carballo et al. – Cuban stingless bees with the specialized resource use6 According to Terrab et al. (2004), pollen grain size is one of the factors influencing higher or lower representation of plant species pollen in honey. Pollen size can be defined by the length of its polar axis and equatorial diameter, and considering the major axis, they are differentiated into several types (Erdtman, 1952). The results indicate that the Cuban Beehives Dominant pollen (>45%) Companion pollen (16-45%) Important isolated pollen (4-15%) Sporadic or rare pollen (1-3%) Present pollen (<1%) 1 Psidium guajava (65,96%) - Mimosa pudica (13,98%) Citrus sp. (6,07%) Alternanthera (2,9%) Moringa oleífera (2,37%) Mimosa pigra (1,58%) Cariofilaceae (1,85%) Acasia farnesiana (0,26%) Trifolium (0,26%) Bidens pilosa (0,53%) Unknown (0,53%) 9 Alternanthera (54,11%) Mimosa pudica (25,44%) Trifolium (7,23%) Psidium guajava (5,49%) Citrus sp. (2,49%) Moringa oleífera (1,5%) Acasia farnesiana (0,75%) Bursera simaruba (0,99%) Bidens pilosa (0,75%) Unknown (0,25%) Mimosa pigra (0,75%) Cariofilaceae (0,25%) 11 Trifolium (55,97%) - Mimosa pudica (14,68%) Alternanthera (10,95%) Acasia farnesiana (4,48%) Bursera simaruba (4,73%) Psidium guajava (2,49%) Citrus sp (1,24%) Moringa oleífera (1,74%) Bidens pilosa (1,74%) Cariofilaceae (1,49%) Albizia lebbeck (0,25%) Unknown (0,25%) 12 Alternanthera (64,79%) - Mimosa pudica (9,26%) Psidium guajava (10,66%) Trifolium (8,25%) Acasia farnesiana (2,21%) Bursera simaruba (1,41%) Moringa oleífera (1,21%) Bidens pilosa (1,81%) Albizia lebbeck (0,2%) Mimosa pigra (0,2%) 13 Alternanthera (58,53%) - Mimosa pudica (9,71%) Acasia farnesiana (6,82%) Trifolium (8,66%) Bursera simaruba (4,2%) Psidium guajava (2,89%) Moringa oleífera (1,05%) Bidens pilosa (1,81%) Albizia lebbeck (1,84%) Citrus sp. (2,36%) Coccoloba uvifera (1,31%) - Table 3. Pollen types identified in the honeys from the beehives of Matanzas, Cuba, 2018. Table 4. Pollen types identified in the honeys from Mayabeque beehives. Beehives Dominant pollen (>45%) Companion pollen (16-45%) Important isolated pollen (4-15%) Sporadic or rare pollen (1-3%) Present pollen (<1%) 14 Mimosa pudica (71,29%) - Psidium guajava (11,88%) Mimosa pigra (4,95%) Cariofilaceae (4,95%) Acasia farnesiana (3,96%) Moringa oleífera (2,97%) - 2 Mimosa pudica (64,11%) Psidium guajava (22,6%) Moringa oleífera (5,52%) Acasia farnesiana (1,54%) Mimosa pigra (2,45%) Bursera simaruba (3,68%) - 48 Mimosa pudica (70,24%) Psidium guajava (23,81%) Mimosa pigra (4,76%) Acasia farnesiana (1,19%) - 5 Mimosa pudica (78,03%) - Mimosa pigra (9,85%) Psidium guajava (6,44%) Acasia farnesiana (1,14%) Moringa oleífera (3,41%) Cariofilaceae (1,14%) - 6 Mimosa pudica (86,6%) - Mimosa pigra (2,9%) Psidium guajava (6,5%) - Acasia farnesiana (0,4%) Moringa oleífera (0,4%) Coccoloba uvifera (0,9%) Arecaceae Tipo Adonidia (0,2%) Nopalia auberi (0,3%) Pristimera coriacea (0,2%) Mangifera indica (0,3%) Myrtaceae Tipo Syzygium (0,6%) Unknown (0,3%) Unknown (0,2%) Sociobiology 69(3): e7729 (September, 2022) 7 stingless bee livestock of both provinces has a predilection for collecting pollen grains of small size (varying from 10 to 25 µm) and explain the highest representation of the species M. pudica in the Mayabeque honey (it measures 10 µm) (Figure 2). The main floral resources used by the livestock of Matanzas province (Fig 3) were, first, M. pudica, with a mean percentage of 55,24%, dominant in three of the five samples, followed by B. simaruba (31,11%) and P. guajava (25,50%). Fabaceae was the most represented family in Matanzas and Mayabeque (Fig 4). Fig 2. Pollen grains of the prevailing types identified in the stingless bee honey and pollen samples of both provinces seen with a magnification of 400. Genus Trifolium (a); Genus Alternanthera (b); Mimosa pudica (c); Bursera simaruba (d) and Psidium guajava (e). Scale bar: 20 µm. Fig 3. Main floral resources identified in the pollen collected from Matanzas province. Dominant pollen (>45%), companion or secondary pollen (16-45%), isolated pollen (4-15%). Fonte-Carballo et al. – Cuban stingless bees with the specialized resource use8 Discussion If the criterion expressed by Pérez de Zabalza (1989) is followed, the studied honey would be classified as of low palynological richness because they represent less than 20 pollen forms. Nevertheless, this is a parameter established for European bee (Apis mellifera Linnaeus, 1758) honey in a Mediterranean environment which largely differs from the tropics. Studies by Imperatriz-Fonseca et al. (1989) and Ramalho (1990) in stingless bees showed some general foraging patterns for Meliponini, proving that, despite foraging on the wide diversity of available floral resources in the area, they are concentrated only on a few species. This would explain the obtained results. Ramalho (2004) observed that the bees of the genus Melipona visit flowers of different shapes, sizes, and colors to obtain their food. However, there was a preference for small flowers with radial symmetry and open corollas in which access to pollen and nectar is relatively easy. Studies conducted in 18 colonies of three species of the Meliponini tribe found a prevalence of small pollen grains, suggesting the preference of these bees for small-size flowers with short pistils (Vossler, 2015). Guimarães et al. (2021) studies confirm the results obtained in the Mayabeque honey regarding the abundance of M. pudica pollen grains (10 µm). These authors detected high representativeness of the Mimosa type in the pollen content of honey samples of Melipona seminigra pernigra Moure and Kerr, 1950, ascribable, according to Ferreira and Absy (2017), to its release in large quantities, their configuration, and the small size of pollen grains (Zappi et al., 2018). In the case of M. beecheii, it has been reported that the type of opening or dehiscence of the anthers (Vossler, 2015) influences the preference for pollen types among the flora in the foraging area. Such preference favors collecting pollen types from Fabaceae, Melastomataceae, and Myrtaceae, which have poricidal openings (Biesmeijer, 1997). Hence, the higher representation of these families in the evaluated honey of both provenances coincides with the observation made by Novais and Absy (2015) and Costa et al. (2017) in the Melionini tribe. The results of the diversity index (H´) for the Matanzas honey (Fig 1) coincide with the value range from 1.23 to 2.00, obtained by Pérez-Sato et al. (2018) in A. mellifera honey produced in the High Plateau of the Puebla State in Mexico. However, they are below the ones reached by Soto (2007) for the stingless bee species Scaptotrigona pectoralis Dalla Torre, 1896, in Guatemala; and the ones reported by Rezende et al. (2020) in three Melipona species in Brazil. For this same index, the Mayabeque provenance was shown in correspondence with those obtained by Novais et al. (2013) for the stingless bee species Tetragonisca angustula Illiger, 1806, in Brazil, with ranges comprised from 0.36 to 2.55. The diversity index (H´) is a tool that determines the breadth of the trophic niche bees through the number of botanical taxa present in the honey samples. Ferreira and Absy (2017) concluded that the breadth of the niche is determined by the movements of massive foraging of bee species with a higher number of workers, which consequently collect more trophic resources. On the other hand, colonies that have less population tend to compete with lower intensity for trophic resources, as we observed for M. beecheii in Matanzas (Table 2). Yet, both localities show a breadth of trophic niches in correspondence with the results of Hilgert-Moreira et al. (2013) for the genus Melipona Illiger, 1806. The evenness index (J´) values were low in Matanzas (0.45) and Mayabeque (0.28). According to Espinoza-Toledo Fig 4. Main floral resources identified in the pollen collected from the Mayabeque province. Dominant pollen (> 45%), companion or secondary pollen (16-45%), isolated pollen (4-15%). Sociobiology 69(3): e7729 (September, 2022) 9 et al. (2018), these results can be explained by the dominance of some species that causes (J´) to show a strong trend toward zero, as observed in the analyzed honey. This fact is markedly proven in the Mayabeque honey (Table 4), where the dominant species in all the samples was Mimosa pudica. In addition, the values reached for this index indicate that the bees from both localities showed oligolectic behavior. According to Biesmeijer (1997), bees classified as oligolectic or specialists behave by visiting and collecting pollen from a very restricted group of plants, which applies to M. beecheii. On the other hand, polylectic or generalist bees utilize pollen and nectar from different plant species in their feeding. In turn, Ramalho et al. (1990) and Rezende et al. (2020) ascribe this characteristic to the fact that the beehives of this stingless bee species are composed of a small number of individuals compared to other species. That is why they focus on few available resources and exploit them intensely. The most represented family in Matanzas and Maybeque (Figure 4) was Fabaceae. Similar results were obtained by Ferreira and Absy (2013) and Barquero-Elizondo et al. (2019) in stingless bee species. They ascribe this fact to the relevance of this family within the ecosystem for its diversity (small-size plants, lianas, shrubs, and trees) and abundant flowerings throughout the year. Within this family (Fabaceae), the species that stood out was Mimosa pudica, reported as a pollen source for bees of the Melipona genus in El Salvador (Biesmeijer, 1997), Colombia (Rodríguez, 2005) and other neotropical habitats. In Cuba, in previous studies conducted by Leal-Ramos and Sánchez (2013), they observed a high frequency of the pollen type of M. pudica in pollen samples obtained from the species M. beecheii. Conclusions A specialist feeding habit characterizes the Cuban stingless bee livestock of the Matanzas and Mayabeque provinces. It has a diet based on a few plant species, among which Bursera simaruba (L.) Sarg., Mimosa pudica L., Psidium guajava L., Trifolium spp., and Alternanthera spp. stand out. In addition, a marked preference for small pollen grains from 10 to 25 µm was observed, especially those of Mimosa pudica in the Mayabeque honey. Acknowledgments The authors acknowledge the support of the Swiss Cooperation and Development Agency (SDC) for its contribution to funding this research through the project BIOMAS-CUBA, in its stage III. Likewise, they thank the Pastures and Forages Research Station Indio Hatuey, the University of Matanzas, and the Palynology and Plant Conservation Group (GPCV) of the Instituto Hispano Luso de Investigaciones Agrarias (CIALE) of the University of Salamanca. Authors’ Contribution LFC: Conceptualization, methodology, investigation, software, validation, formal analysis, resources, data curation, writing- original draft, writing-review & editing, visualization, supervisión, project administration. WLP: Conceptualization, investigation, resources JDL: Conceptualization, investigation, resources MDS: Conceptualization, investigation, resources DMQ: Conceptualization, investigation, software, resources. DRC: Methodology, investigation, software, validation, formal análisis, resources, writing-original draft, writing-review & editing. JSS: Methodology, investigation, validation, resources. ESR: Methodology, investigation, validation¸ resources. References Academia de Ciencias de Cuba. (1989). Nuevo Atlas Nacional de Cuba. Instituto Cubano de Geodesia y Cartografía. La Habana, Cuba. p. 41. Aira, M.J., Chávez, M.A., Fernández-González, M. & Rodríguez-Rajo, F.J. (2018). Pollen diversity in the atmosphere of Havana, Cuba. Aerobiologia, 34: 389-403. Araújo, E.D., Costa, M., Chaud-Netto, J. & Fowler, H.G. (2004). Body size and flight distance in stingless bees (Hymenoptera: Meliponini): inference of flight range and possible ecological implications. Brazilian Journal of Biology, 64: 563-568. Barquero-Elizondo, A.I., Aguilar-Monge, I., Méndez-Cartín, A.L., Hernández-Sánchez, G., Sánchez-Toruño, H., Montero- Flores, W. & Mesén-Montano, I. (2019). Asociación entre abejas sin aguijón (Apidae, Meliponini) y la flora del bosque seco en la región norte de Guanacaste, Costa Rica. Revista de Ciencias Ambientales, 53: 70-91. Biesmeijer, J.C. (1997). Abejas sin aguijón. Su biología y organización de la colmena. Elinkwijk BV, Utrecht. Costa, C.C.A., Silva, C.I., Maia-Silva, C. Limão, A.A.C. & Imperatriz-Fonseca, V.L. (2017). Origem botânica do mel da jandaíra em áreas de caatinga nativa do Rio Grande do Norte. In: V.L. Imperatriz-Fonseca, D. Koedam, M. Hrncir (Eds.), A abelha jandaíra no passado, no presente e no futuro (pp. 161- 166). Mossoró: Ed. Ufersa. Erdtman, G. (1952). Pollen morphology and plant taxonomy. Angiosperms. Stockholm: Almqvist and Wiksell, 539 p. Espinoza-Toledo, C., Vázquez-Ovando, A., Torres de los Santos, R., López-García, A., Albores-Flores, V. & Grajales- Conesa J. (2018). Stingless bee honeys from Soconusco, Chiapas: a complementary approach. Revista de Biología Tropical, 66: 1536-1546. Ferreira, M.G. & Absy, M.L. (2013). Pollen analysis of the post-emergence residue of Melipona (Melikerria) interrupta Fonte-Carballo et al. – Cuban stingless bees with the specialized resource use10 Latreille (Hymenoptera: Apidae) bred in the central Amazon region. Acta Botanica Brasilica, 27: 709-713. Ferreira, M.G. & Absy, M.L. (2017). Pollen analysis of honey of Melipona (Michmelia) seminigra merrillae and Melipona (Melikerria) interrupta (Hymenoptera: Apidae) bred in Central Amazon, Brazil. Grana, 56: 436-449. Flores, F.F., Lupo, L.C. & Hilgert, N.I. (2015). Recursos tróficos utilizados por Plebeia intermedia (Apidae, Meliponini) en la localidad de Baritú, Salta, Argentina. Caracterización botánica de sus mieles. Boletín de la Sociedad Argentina de Botánica, 50: 515-529. Fonte, L. (2007). Caracterización Preliminar de las “abejas de la tierra” y sus “tenedores”, las colmenas y la miel que producen, en zonas de las provincias occidentales de Cuba. Degree work. Facultad de Medicina Veterinaria. La Habana: Universidad Agraria de la Habana., 81 p. García, M., Pérez-Arquillue, C., Juan, T., Juan, M.I. & Herrera, A. (2001). Note. Pollen analysis and antibacterial activity of Spanish honey. Food Science and Technology International, 7: 155-158. García-Abad, J.J. (2015). Abundancia relativa, frecuencia y riqueza de plantas vasculares a escala local. Metodología de Índices de Ocupación de la Flora (aplicada a La Alcarria Occidental). Estudios Geográficos, 76: 499-530. González-Porto, A.V., Sánchez Reyes E., De Linares Fernández C., Rodríguez de la Cruz D., Sánchez Sánchez J., Belmonte Soler J., Seijo Coello M.C., Valencia Barrera R.M., García Rogado M.R. & Matías Martínez Y. (2018). El Grupo de Trabajo de la Miel y la normalización de protocolos de análisis melisopalinológicos (comunicación oral). IX Congreso Nacional de Apicultura (Tenerife, España). Guimarães, J.T.F., Costa, L., Zappi, D.C., Batista Junior, W.F., Lopes, K.D.S., Alves, R.C.D.O. & Barth, O.M. (2021). Preferências de forrageamento da abelha nativa sem ferrão Melipona seminigra pernigra (Apidae: Meliponini) em campo rupestre na canga da Serra dos Carajás, sudeste da Amazônia. Biota Neotropica, 21: e20201004 Henderson, P.A. & Seaby, R.H. (2002). Species Diversity and Richness 3.02. Lymington, UK: Pisces Conservation Ltd, http://www.pisces-conservation.com. [12/03/18]. Hilgert-Moreira, S.B., Nascher, C.A., Callegari-Jacques, S.M. & Blochtein, B. (2013). Pollen resources and trophic niche breadth of Apis mellifera and Melipona obscurior (Hymenoptera, Apidae) in a subtropical climate in the Atlantic rain forest of southern Brazil. Apidologie, 45: 129‒141. Imperatriz-Fonseca, V.L., Kleinert-Giovannini, A. & Ramalho, M. (1989). Pollen harvest by eusocial bees in a non-natural community in Brazil. Journal of Tropical Ecology, 5: 239-242. Leal-Ramos, A. & Sánchez, L.E (2013). Antagonismo de Apis mellifera y Melipona beecheii por las fuentes de alimentación. Revista Cubana de Ciencias Forestales 1:102-109. Lóriga, W. (2015). Caracterización de las abejas, colmenas, sistema de manejo y estado de salud de Melipona beecheii Bennett (Apidae, Meliponini) en áreas del Occidente de Cuba. Doctoral Thesis. San José de las Lajas: Facultad de Medicina Veterinaria, Universidad Agraria de La Habana, 98 p. Louveaux J., Maurizio A. & Vorwhol G. (1978). Methods of Melissopalinology. Bee World, 59: 139-157. Novais, J.S., Absy, M.L. & dos Santos, F.D.A.R. (2013). Pollen grains in honeys produced by Tetragonisca angustula (Latreille, 1811) (Hymenoptera: Apidae) in tropical semiarid areas of northeastern Brazil. Arthropod-Plant Interactions, 7: 619-632. Novais, J.S. & Absy, M.L. (2015). Melissopalynological records of honeys from Tetragonisca angustula (Latreille, 1811) in the Lower Amazon, Brazil: Pollen spectra and concentration. Journal of Apicultural Research, 54: 11-29. Nunes-Silva, P., Costa, L., Campbell, A.J., Arruda, H., Contrera, F.A.L., Teixeira, J.S.G. & Imperatriz-Fonseca, V.L. (2020). Radiofrequency identification (RFID) reveals long-distance flight and homing abilities of the stingless bee Melipona fasciculata. Apidologie, 51: 240-253. Pielou, E.C. (1984). The interpretation of ecological data. New York: John Wiley and Sons, Inc., 262 p. Pérez, K. (2016). Determinación del origen botánico de mieles de Melipona beecheii Bennett (Apidae, Meliponini) en meliponarios de la provincia de Mayabeque. Degree work. La Habana: Facultad de Agronomía. Universidad Agraria de La Habana, 51 p. Pérez-Piñeiro, A. (2017). La apicultura en cuba y su situación actual. Agroecología, 12: 67-73. Pérez-Sato, M., Flores-Garrido, A.F., Castro-González, N.P., Escobar-Hernández, R., Soni-Guillermo, E. & Pérez-Hernández, H. (2018). Análisis palinológico de la miel de Apis mellifera L., producida en el Altiplano del estado de Puebla, México. Guía para autores. AGRO Productividad, 11: 98-104. Pérez de Zabalza, A. (1989). Estudio palinológico de las mieles de Navarra. Doctoral Thesis. Navarra: Universidad de Navarra. 268 p. RAE. (2017). Ganado. Madrid: Real Academia Española. http://dle.rae.es/?id=IpmVKGf. Ramalho, M. (1990). Foraging by stingless bees of the genus Scaptotrigona (Apidae, Meliponinae). Journal of Apicultural Research, 29: 61 -67. Ramalho, M., Kleinert-Giovannini, A. & Imperatriz-Fonseca, V. L. (1990). Important bee plants for stingless bees (Melipona http://www.pisces-conservation.com Sociobiology 69(3): e7729 (September, 2022) 11 and Trigonini) and Africanized honeybees (Apis mellifera) in neotropical habitats: a review. Apidologie, 21: 469-488. Ramalho, M. (2004). Stingless bees and mass flowering trees in the canopy of Atlantic Forest: a tight relationship. Acta Botanica Brasilica, 18: 37-47. Rezende, A.C.C., Absy, M.L., Ferreira, M.G. & Marinho, H.A. (2020). Honey botanical origin of stingless bees (Apidae Meliponini) in the Nova América community of the Sateré Mawé indigenous tribe, Amazon, Brazil. Grana, 59: 304-318. Rodríguez, A. (2005). Estudio del forrajeo de polen por obreras de Melipona fasciata (Hymenoptera, Apidae, Meliponini) en el piedemonte llanero colombiano (Acacías-Meta-Colombia). IV Seminario Mesoamericano sobre abejas sin aguijón. Chalatenango, El Salvador. Sáenz, C. & Gómez, C. (2000). Mieles españolas. Características e identificación mediante el análisis de polen. Madrid: Ed. Mundi prensa. 129 p. Sánchez, L. (2001). Métodos palinológicos. Curso de capacitación. PROMABOS. San Salvador, El Salvador. Sánchez, E., Rodríguez, D., Sánchez, S., Vega, E. & Sánchez, J. (2018). Sobre la metodología en los estudios melisopalinológicos. Coimbra: V Congreso Ibérico de Apicultura. Shannon C.E. & Weaver W. (1949). The mathematical Theory of Communication. University of Illinois Press, 117 p. Soto, M.A.V. (2007). Recursos polínicos utilizados por la abeja nativa Shuruya (Scaptotrigona pectoralis) (Apidae: Meliponini) en un meliponario de la parte baja de los cipresales em Pachalum, Quiché, durante la época seca y lluviosa. Doctoral Thesis. Facultad de Ciencias Químicas y Farmacia, Universidad de San Carlos de Guatemala. 73 p. Terrab, A., Pontes, A., Heredia FJ. & Diez, M.J. (2004). Palynological and geographical characterization of avocado honeys in Spain. Grana, 43: 116-121. Vossler, F.G. (2015). Small pollen grain volumes and sizes dominate the diet composition of three South American subtropical stingless bees. Grana, 54: 68-81. Wittig, R. & Becker, U. (2010). The spontaneous flora around street trees in cities – A striking example for the worldwide homogenization of the flora of urban habitats. Flora- Morphology, Distribution. Functional Ecology of Plants, 205: 704-709. Zappi, D.C., Gastauer, M., Ramos, S., Nunes, S., Caldeira, C. F., Souza-Filho, P.W., Guimarães, J.T.F., Giannini, T.C., Viana, V.P.L., Lovo, J., Mota, N.F.O. & Siqueira, J.O. (2018). Plantas nativas para recuperação de áreas de mineração em Carajás. Belém, PA: Instituto Tecnológico Vale (ITV). 282 p.