Open access journal: http://periodicos.uefs.br/ojs/index.php/sociobiology ISSN: 0361-6525 Sociobiology 60(1): 56-64 (2013) The Botanical Profiles of Dried Bee Pollen Loads Collected by Apis mellifera (Linnaeus) in Brazil AS Freitas1, VAS Arruda2, LB Almeida-Muradian2, OM Barth1,3 Introduction The marketing for Apis mellifera (L). products is widely expanding in Brazil in response to the increased exports in recent years. Bee pollen production has gradu- ally increased in response to the demand for dietary supple- ments and therapeutic products (Barreto et al. 2005). These products result from the intensive pollination activity of the bees, which support the maintenance of the biodiversity of the pollen-producing flora (Santos 2009). Beekeeping has been widespread in several Brazilian regions as an activ- ity that supplements the income of small farmers while it has a low environmental impact and high importance for the social and the economic aspects of sustainability (Vieira & Resende, 2007). Brazilian vegetation is geographically diverse and is divided into five macro-regions. The North region includes the largest tropical rainforest in the country such as a huge drainage area through the Amazon basin. The Northeast re- Abstract A total of 61 dried bee pollen samples collected in four Brazilian macro-regions within 19 municipalities were analyzed aiming to identify the sources used by Apis mellifera (L.) for pollen production and to enable a more accurate product certification. Sample preparation followed the standard methodology, including washing the pollen grains with ethanol, then with water and homogenising the sediment in a water/glycerine solution for microscopic observation. Pollen counts included at least 500 pollen grains per sample. Only six samples, presenting a unique species or pollen type comprising more than 90% of the pollen sum, were considered monofloral, including Ambrosia sp., Cecropia sp., Eucalyptus sp., Fabaceae, Mimosa scabrella (Benth.) spp. and Schinus sp. pollen types. The most frequent pollen types of the heterofloral pollen batches, based on a counting limit of 45%, included Anadenanthera sp., Asteraceae, Brassica sp., Caes- alpiniaceae, Cocos nucifera (L.) sp., Mimosa caesalpiniaefolia (Benth.) sp., Mimosa ver- rucosa (Benth.) sp., and Myrcia sp. pollen types. This result may be related to the great diversity of the Brazilian flora contributing to heterofloral pollen loads and honeys. Sociobiology An international journal on social insects 1 - Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil. 2 - Universidade de São Paulo, São Paulo, Brazil. 3 - Fundação Oswaldo Cruz, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil ReSeARCh ARTiCLe - BeeS Article History edited by: isabel Alves-dos-Santos, USP - Brazil Received 25 December 2012 initial acceptance 27 February 2013 Final acceptance 06 March 2013 Keywords palynology, bee flora, pollen batches, Brazil Corresponding author Alex da Silva de Freitas Universidade Fed. do Rio de Janeiro Laboratório de Palinologia Departamento de Geologia, CCMN Rio de Janeiro, RJ, Brazil e-Mail: alexsilfre@gmail.com gion occurs in the eastward “bulge” of the country where there is a semiarid interior been largely given over to low- density vegetation. The Southeast region is mainly an upland area with prevalence of high species diversity in the vegeta- tion of the Atlantic forest. The South region is the smallest macro-region, been distinct because of its temperate climate and the presence of Araucaria forest. In the Midwest region there is a dry forest (“cerrado”) characterized by drier cli- mate and vegetation resistant to prolonged periods without rain, besides the humid “pantanal” areas (Rizzini 1997). For many insects, and particularly for bees, pollen is the main source of the proteins and lipids required by lar- vae. Pollen is essential to the normal growth and develop- ment of all individuals in a colony and for the reproduction and maintenance of the colony (Marchini et al. 2006). Pol- len grains are collected on the corbicula of the hind legs of the bees during field activity and are stored in hive cells, forming the “bee bread” (Moreti et al. 2002). The increasing variety and availability of apiary derivatives in the Brazilian Sociobiology 60(1): 56-64 (2013) 57 market, and international interest in these products encour- age research to characterize the botanical origin of different pollen types collected by bees using palynological analysis for quality control (Luz et al. 2007). The botanical origin of bee pollen loads is of extreme importance to a greater quality control, enabling better com- mercial value for the manufactured products. Almeida-Mu- radian et al. (2005) emphasized the importance of this qual- ity control in targeting the commercial production of pollen products, including the flowering plants of each region and a wide range of physicochemical bee pollen analyses. Melissopalinological analysis is an important tool for increasing knowledge about the botanical origins of bee pol- len. Regional data were presented by Barth & Luz (1998) with regard to pollen collection inside mangrove areas, Barth (2004) in a review, Modro et al. (2007), Luz et al. (2007) in a forested area of Rio de Janeiro State, Melo et al. (2009) in São Paulo State, Barth et al. (2009), Luz et al. (2010) in Minas Gerais State, Barth et al. (2010) with regard to the technical processing of pollen loads, Luz et al. (2011) in Espírito Santo State, Boff et al. (2011) in Mato Grosso do Sul State, Novais et al. (2009); Novais et al. (2010); Dórea et al. (2010); Santos (2011) in Bahia State and Barth et al. (2011) in regions of Venezuela. The present study aimed to investigate the botani- cal and geographical origins of bee pollen collected in four Brazilian macro-regions. This study aimed to provide also results that will contribute to improve the certification of this bee product and the preservation of the native vegetation. Materials and Methods A total of 61 dried pollen load samples of A. mellif- era collected by traps in four Brazilian macro-regions, were analyzed using pollen analysis (Figure 1). The procedure followed the methodology proposed by Barth et al. (2010) using two grams of each sample of dried bee pollen homoge- nate stirring in 70% ethanol. Centrifuge tubes were filled to 13 mL, and the material remained at rest for 30 min or over- night and was then submitted to sonication for five minutes to dissociate the pollen grains. After centrifugation, the samples containing a large amount of oil were submitted twice to ethanol extraction. The obtained sediment was diluted in a 1:1 mixture of water/ glycerine for 30 min. One drop of this well-homogenised pollen grain suspension was applied to a microscope slide and covered with a 24x24 mm cover glass. The stock pollen suspension was kept in glycerine at room temperature. Two microscope slides, sealed with nail-lack, were prepared and more than 500 pollen grains were counted from each sample considering the two slides. Samples were observed using light and polarized light microscopy. Pollen Figure.1 Map of the five Brazil- ian macro-region displaying the studied area with the municipali- ties collection. 1. Rio Grande do Norte; 2. Sergipe; 3. Bahia; 4. Espírito Santo; 5. São Paulo; 6. Santa Catarina; 7. Rio Grande do Sul; 8. Distrito Federal; 9. Mato Grosso. AS Freitas, VAS Arruda, LB Almeida-Muradian, OM Barth - Botanical profiles of pollen loads collected by Apis mellifera58 classes usually followed those established by Zander (Lou- veaux et al. 1978) meaning honey analysis PP (predominant pollen > 45%), AP (accessory pollen 15-45%), IP (important pollen 3-15%). These classes were used for qualitative and quantitative analyses of bee pollen loads in the present pa- per. Samples were classified as monofloral or hetero- floral batches according to their pollen grain percentages. Samples were considered monofloral when presenting more than 90% of a unique pollen type. Barth (1989), Roubik & Moreno (1991) and Moreti et al. (2002) were used to aid in pollen identification. Pollen morphology in the Asteraceae family is not genus specific and was presented at the family ranking Asteraceae. Results The results of the palynological analysis showed a wide variety of pollen types found in each Brazilian macro- region (Tables 1, 2, 3). Six of the 61 samples were monoflo- ral. The monofloral samples were found in two samples from the Southeast, two from the South and two from the Midwest (Tables 2 and 3). Northeast region (Table 1) – Pollen from Cocos nu- cifera was present in all 19 samples analyzed, with a con- centration above 3%. This pollen type was present as a dominant pollen in seven samples (>45%), as an accessory pollen in ten samples and as an isolate pollen in two samples throughout the three states of the Northeast region. Pollen types of Mimosa caesalpiniaefolia. (sample Sergipe-7), M. scabrella (samples Bahia-2, -5), M. verrucosa (samples Ser- gipe-5, -6, -8, -10, -11), and Myrcia sp. (samples Sergipe-2, -3, -11) also exhibited a dominant pollen frequency. Southeast region (Table 2) – The unique dried bee pollen sample from Espírito Santo State was a monofloral sample of an unidentified plant of the Fabaceae family was only found in that one sample. Nine samples from Pariquera Açu in São Paulo State all came from Cocos nucifera and were very similar. The Ribeirão Preto municipality presented dominant pollen grain percentages of M. caesalpiniaefolia in six sam- ples (São Paulo-10, -11, -12, -13, -14, -15). Schinus sp. was the main pollen type in two samples (São Paulo-16, -18) and Anadenanthera sp. in one sample only (São Paulo-19). The other two samples (São Paulo-17, -20) exhibited no domi- nant pollen types but had in common the pollen types of An- adenanthera sp., Eucalyptus sp., Myrcia sp., and Cecropia sp. Pollen grains of the Persea sp. type (avocado tree) were detected in only one sample (São Paulo-17). Taubaté municipality exhibited the M. caesalpini- aefolia (samples São Paulo-21, -23) and M. scabrella (São Paulo-22, -24) pollen types as dominant in two samples each, with the latter including a monofloral sample (São Paulo-24). South region (Table 3) – All samples analyzed from Fraiburgo municipality (Santa Catarina-1, -2, -3, -4, -5, -6) were very similar, presenting a high percentage of Aster- aceae pollen grains as well as Amaranthus/Chenopodiaceae, Brassica sp. and Euphorbiaceae pollen grains in several samples. A Caesalpiniaceae pollen type was dominant in Campos Novos (Santa Catarina-7), and M. caesalpiniaefolia was dominant in Serra Catarinense (Santa Catarina-8). Five samples were analyzed from Rio Grande do Sul State. Three of these samples showed Eucalyptus sp. pollen dominance in the municipalities of Arvorezinha (Rio Grande do Sul-4), Santana do Livramento (Rio Grande do Sul-2) and São Gabriel (Rio Grande do Sul-5). The Ambrosia sp. pollen type occurred in a sample from Santana do Livra- mento (Rio Grande do Sul-1), and the Brassica sp. pollen type was found in a sample from Cruz Alta (Rio Grande do Sul-3), both of which were classified as monofloral. Midwest region (Table 3) - Midwestern Brazil was represented by four samples. The Distrito Federal sample showed a high percentage of the Schinus sp. pollen type, which was considered monofloral. Three samples from Mato Grosso showed Cecropia sp. pollen dominance, one of which (Mato Grosso-2) was considered monofloral. The most frequent or characteristic pollen types in the Brazilian macro-regions of the present study can be sum- marised as follows: Cocos nucifera (n = 19/19) is a typical pollen product in the Brazilian Northeast region (n = 19) and, pollen types of M. caesalpiniaefolia (n = 3), M. scabrella (n = 8) and M. verrucosa (n = 7) were common. Pollen grains of Aster- aceae species were poorly represented in the samples of this macro-region. The main pollen types of the Southeast region (n = 25) were Asteraceae and Myrcia sp. (n = 11/25 of each), such as M. caesalpiniaefolia (n = 12/25). The characteristic pollen types of the South macro-region (n = 13) were Asteraceae (n = 10/13) and Brassica sp. (n = 8/13). In the few samples of the Midwest macro-region (n = 4) Cecropia sp. (n = 4) pol- len was detected in all samples, including in one monofloral sample primarily of Schinus sp. pollen (n = 1). Discussion Northeast region – Coconut trees grow frequently on seashores along the northeastern Brazilian coastline (Pires et al. 2004) and are naturally spread throughout the beaches. These trees are considered an important pollen source in Neotropical regions (Biesmeijer et al. 1992). Cocos nucifera pollen grains were well represented in all samples of the three Northeast states (Rio Grande do Norte, Sergipe and Bahia). The pollen of this species was associated with M. verrucosa (“espinheiro preto”), a typical plant throughout the Brazilian Northeast (Sodré et al. 2007), and mainly in samples of the state of Sergipe. In addition, M. caesalpiniaefolia (“sabiá”) Sociobiology 60(1): 56-64 (2013) 59 was found in association with Cocos nucifera. Novais et al. (2009) and Dórea et al. (2010) highlight other Mimosacea and Arecaceae species for the Bahia State. M. caesalpiniaefolia has a natural occurrence in the semi-arid Brazilian region (Carvalho 2007, Queiroz, 2009) and has been introduced in more humid regions (Ribaski et al. 2003) and is widely used in the reforestation of degraded areas (Lorenzi 1998). Caesalpiniaceae has been reported by several authors (Absy & Kerr, 1977; Absy et al. 1980; Kerr et al. 1986/1987; Imperatriz-Fonseca 1989; Ramalho 1990) as an important food resource for native bees in different Brazilian states. Monofloral pollen loads of M. caesalpini- aefolia have also been collected in the state of Rio de Janeiro (Luz et al. 2007). Therefore it may be considered that the pollen of Cocos and Mimosa is very important in the North- east Brazilian region. Southeast region - The Fabaceae family is often vis- ited by bees in search of nectar and pollen for the mainte- nance of the hive (Moreti et al. 2007). A monofloral pollen batch of a Fabaceae species came from Espírito Santo State. This species may be wild or a crop species, and the pollen morphology of several related species similar (Carreira et al. 1996; Moreti et al. 2007). The Mimosaceae family compris- es a large number of species of the Brazilian flora that are of extreme significance for pollen and honey production. One batch with predominant pollen of the M. scabrel- la pollen type was obtained in the State of São Paulo. The pollen grain morphology of this specie is not characteristic of a unique plant species but rather, groups of species (Barth 1989). Based on a survey of Neotropical region, Ramalho et al. (1990) reported that the M. scabrella pollen type was commonly collected by A. mellifera. The numerous monofloral batches of the Cocos nu- cifera pollen type obtained in the state of São Paulo were probably derived from the extensive coconut plantations in the region. Surprisingly, these pollen grains dominated all samples from the munipality of Pariquera Açu, São Paulo State, indicating that coconut plantations are significant for beekeeper activities. The attractive flowering of the wild trees or shrubs of Anadenanthera sp. is extremely important for beekeep- ers as because, in addition to pollen and honey harvesting, these flowering plants draw hungry bees away from crop plantations, as of Passiflora sp. such as those pollinated by bumblebees, where bees are not welcome (Monika Barth, personal information). South region - The Ambrosia sp. pollen type was characteristic of the state of Rio Grande do Sul (RS). The anemophilous pollen of this genus, dispersed by the wind, is considered allergenic in this region and is suspected to have induced hay fever in several patients (Vergamini et al. 2006). According to Lorenzi (2008), species of the Ambro- sia genus have been found in agricultural areas of southern Brazil and are considered a weed species. This pollen type is visited by a variety of insects. A monofloral pollen batch of Eucalyptus sp. was also obtained in Rio Grande do Sul state, but this genus is com- mon throughout the Brazilian regions where it is cultivated. Pollen analyses of honey have certified the presence of Eu- calyptus sp. (Barth et al. 2005; Bastos et al. 2004; Luz et al. 2007). The pollen morphology of Brassica sp. which is similar in several species may be derived from crops (Bras- sica napus L.) in the south of Brazil (Rosa et al. 2011) or from wild species occurring in field vegetation (Moreti et al. 2002). Midwest region - Another monofloral batch of wind- dispersed pollen was Cecropia sp. from the region of Mato Grosso do Sul. Cecropia is a widespread small tree in Brazil and produces small pollen that is very attractive to bees (Luz et al. 2007). This genus was also observed by Boff et al. (2011) as an accessory pollen type in analyzed pollen loads from southern Pantanal in the state of Mato Grosso do Sul (MS). Bees have a “preference” for Schinus sp. pollen. The main species, S. terebinthifolius (L.), has a worldwide dis- tribution in tropical regions. The present monofloral sample was obtained in the Brazilian Federal District. The pollen of this genus occurs as an accessory percentage of the total pol- len samples, including in honey samples (Carreira & Jardim, 1994). This genus is characteristic of drier vegetation (Smith et al. 2004) and is widely sought by bees, especially when other species are not in the flower (Baggio 1988). All dominant pollen types were present as accessory pollen in numerous samples. In addition, six non-dominant pollen types were identified as accessory pollen in the pres- ent study including pollen types of Anadenanthera sp., Brassica sp., Caesalpiniaceae, Cocos nucifera, M. caesal- piniaefolia and Myrcia sp. Several plant species and genera showed very similar pollen morphology to that of Myrcia sp. (Barth 1972). This pollen type occurs in pollen batches of nearly all Brazilian states and was also highlighted by Barth (1989) as an acces- sory pollen type in honey samples from Bahia. From the data presented here, the great floristic diver- sity in Brazil is apparent, as shown through the main pollen types identified in bee pollen loads. Thus, there is a need for a comprehensive and detailed botanical certification pro- gram to increase the commercial value of pollen products. Acknowledgements We thank the Conselho Nacional de Desenvolvi- mento Científico e Tecnológico (CNPq) and the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) for financial support and scholarships to the three last authors. Two anonymous reviwers provided valuable comments that improved the article. AS Freitas, VAS Arruda, LB Almeida-Muradian, OM Barth - Botanical profiles of pollen loads collected by Apis mellifera60 Table 1. The pollen types identified in the pollen load samples of A. mellifera collected in Northeast macro-region of Brazil and their main botanical origins, considering a frequency of more than 3%. (Samples were considered monofloral when presenting more than 90% of a unique pollen type). Municipality Brazilian Macro-region = Northeast Pollen analysis and pollen types Predominant botanical origin (representative pollen types) Rio Grande do Norte-1 (Natal) PP: Cocos nucifera (60.2%) AP: Mimosa verrucosa (36.0%) Heterofloral sample with primary contributions from Cocos nucifera and Mimosa verrucosa Rio Grande do Norte-2 (Gru- po Santa Luzia) PP: Cocos nucifera (47.2%) AP: Eucalyptus (25.0%) IP: Mimosa caesalpiniaefolia (6.4%), Myrcia (5.2%), Mimosa scabrella (3.7%), Asteraceae (3.4%), Richardia (3.2%) Heterofloral sample with primary contributions from Cocos nucifera and Eucalyptus Sergipe-1 (Povoado Brejão) AP: Mimosa scabrella (42.1%), Cocos nucifera (35.4%), Myrcia (17.5%) Heterofloral sample with primary contributions from Mimosa scabrella, Cocos nucifera and Myrcia Sergipe-2 (Povoado Brejão) PP: Myrcia (67.0%) IP: Cocos nucifera (3.5%) Heterofloral sample with a primary contribution from Myrcia Sergipe-3 (Povoado Brejão) PP: Myrcia (55.5%) AP: Mimosa scabrella (25.3%), Cocos nucifera (17.2%) Heterofloral sample with primary contributions from Myrcia, Mimosa scabrella and Cocos nucifera Sergipe-4 (Povoado Brejão) PP: Cocos nucifera (47.5%) AP: Myrcia (23.2%), Mimosa scabrella (23.2%) Heterofloral sample with primary contributions from Cocos nucifera, Myrcia and Mimosa scabrella Sergipe-5 PP: Mimosa verrucosa (50.7%) AP: Cocos nucifera (41.1%) IP: Poaceae (8.0%) Heterofloral sample with primary contributions from Mimosa verrucosa and Cocos nucifera Sergipe-6 AP: Mimosa verrucosa (42.8%), Cocos nucifera (35.4%), Myrcia (16.3%) IP: Tapirira (5.4%) Heterofloral sample with primary contributions from Mimosa verrucosa, Cocos nucifera and Myrcia Sergipe-7 PP: Mimosa caesalpiniaefolia (50.1%) AP: Cocos nucifera (41.3%) IP: Myrcia (5.7%) Heterofloral sample with primary contributions from Mimosa caesalpiniaefolia and Cocos nucifera Sergipe-8 AP: Mimosa verrucosa (39.7%), Cocos nucifera (36.8%), Mimosa caesalpiniaefolia (15.7%) IP: Myrcia (7.8%) Heterofloral sample with primary contributions from Mimosa verrucosa, Cocos nucifera and Mimosa caesalpiniaefolia Sergipe-9 AP: Cocos nucifera (35.1%), Brassica (20.3%), Myrcia (18.6%), Mimosa verrucosa (16.5%) IP: Tapirira (9.4%) Heterofloral sample with primary contributions from Cocos nucifera, Brassica, Myrcia and Mimosa verrucosa Sergipe-10 PP: Mimosa verrucosa (56.0%) AP: Cocos nucifera (29.3%) IP: Brassica (14.8%) Heterofloral sample with primary contributions from Mimosa verrucosa and Cocos nucifera Sergipe-11 PP: Cocos nucifera (53.3%); AP: Mimosa verrucosa (39.9%) IP: Myrcia (3.8%) Heterofloral sample with primary contributions from Cocos nucifera and Mimosa verrucosa Bahia-1 (Canavieiras) PP: Cocos nucifera (67.6%) AP: Eucalyptus (31.3%) Heterofloral sample with primary contributions from Cocos nucifera and Eucalyptus Bahia-2 (Canavieiras) PP:Mimosa scabrella (59.0%) AP: Cocos nucifera (41.0%) Heterofloral sample with primary contributions from Mimosa scabrella and Cocos nucifera Bahia-3 (Canavieiras) PP: Cocos nucifera (76.3%) AP: Mimosa scabrella (18.4%) IP: Eucalyptus (5.3%) Heterofloral sample with primary contributions from Cocos nucifera and Mimosa scabrella Bahia-4 (Canavieiras) PP: Cocos nucifera (85.5%) IP: Vernonia (6.9%) Heterofloral sample with a primary contribution from Cocos nucifera Bahia-5 (Ilhéus) PP: Mimosa scabrella (58.3%) AP: Cocos nucifera (32.7%) IP: Asteraceae (6.7%) Heterofloral sample with primary contributions from Mimosa scabrella and Cocos nucifera Bahia-6 (Ilhéus) PP: Asteraceae (81.5%) IP: Cecropia (6.4%), Mimosa scabrella (5.3%), Cocos nucifera (4.5%) Heterofloral sample with a primary contribution from Asteraceae PP = predominant pollen (> 45%); AP = accessory pollen (15-45%); IP = isolate pollen (3-15%). Sociobiology 60(1): 56-64 (2013) 61 Table 2. The pollen types identified in the pollen load samples of A. mellifera collected in Southeast macro-region of Brazil and their main botanical origins, considering a frequency of more than 3%. (Samples were considered monofloral when presenting more than 90% of a unique pollen type.) Municipality Brazilian macro-region = Southeast Pollen analysis and pollen types Predominant botanical origin (representative pollen types) Espírito Santo PP: Fabaceae- Faboideae (90.6%) IP: Myrcia (4.7%), Eucalyptus (3.1%) Monofloral sample of a Fabaceae- Faboideae species São Paulo-1 (Pariquera Açu) PP: Cocos nucifera (62.5%) AP: Asteraceae (15.6%) IP: Poaceae (12.5%), Melastomataceae (9.4%) Heterofloral sample with primary contributions from Cocos nucifera, and Asteraceae São Paulo-2 (Pariquera Açu) PP: Cocos nucifera (50.0%) AP: Myrcia (22.2%), Mimosa caesalpiniaefolia (19,4%) IP: Mimosa scabrella (8.3%) Heterofloral sample with primary contributions from Cocos nucifera, Myrcia and Mimosa caesalpiniaefolia São Paulo-3 (Pariquera Açu) PP: Cocos nucifera (55.3%) AP: Mimosa caesalpiniaefolia (27.8%), Myrcia (16.9%) Heterofloral sample with primary contributions from Cocos nucifera Mimosa caesalpiniaefolia and Myrcia São Paulo-4 (Pariquera Açu) PP: Cocos nucifera (47.5%) AP: Myrcia (17.0%), Cecropia (16.4%) IP: Mimosa caesalpiniaefolia (13.8%) Heterofloral sample with primary contributions from Cocos nucifera, Myrcia, and Cecropia São Paulo-5 (Pariquera Açu) PP: Cocos nucifera (81.2%) IP: Ilex (6.2%), Sebastiania (6.2%), Asteraceae (5.3%), Myrcia (4.2%), Heterofloral sample with a primary contribution of Cocos nucifera São Paulo-6 (Pariquera Açu) PP: Cocos nucifera (78.3%) IP: Ilex (11.6%), Asteraceae (5.8%), Myrcia (4.3%) Heterofloral sample with a primary contribution from Cocos nucifera São Paulo/SP 7 (Pariquera Açu) PP: Cocos nucifera (57.7%) IP: Cecropia (12.1%), Ilex (9.1%), Eucalyptus (4.8%), Asteraceae (7.0%), Myrcia (5.8%), unidentified (3.0%) Heterofloral sample with a primary contribution from Cocos nucifera São Paulo-8 (Pariquera Açu) PP: Cocos nucifera (68.3%) IP: Eucalyptus (7.6%), Myrcia (3.8%) Heterofloral sample with a primary contribution from Cocos nucifera São Paulo-9 (Pariquera Açu) PP: Cocos nucifera (58.2%) AP: Cecropia (23.9%), Mimosa caesalpiniaefolia (17.9%) IP: Myrcia (3.8%) Heterofloral sample with primary contributions from Cocos nucifera, Cecropia and Mimosa caesalpiniaefolia São Paulo-10 (Ribeirão Preto) PP: Mimosa caesalpiniaefolia (57.9%) AP: Asteraceae (26.8%) IP: Poaceae (14.6%) Heterofloral sample with primary contributions from Mimosa caesalpiniaefolia and Asteraceae São Paulo-11 (Ribeirão Preto) PP: Mimosa caesalpiniaefolia (60.3%) AP: Asteraceae (17.6%), Poaceae (16.6%), IP: Myrcia (3.8%) Heterofloral sample with primary contributions from Mimosa caesalpiniaefolia, Asteraceae and Poaceae São Paulo-12 (Ribeirão Preto) PP: Mimosa caesalpiniaefolia (70.2%) AP: Poaceae (18.2%) IP: Asteraceae (11.6%) Heterofloral sample with primary contributions from Mimosa caesalpiniaefolia and Poaceae São Paulo-13 (Ribeirão Preto) PP: Mimosa caesalpiniaefolia (51.3%) AP: Mimosa scabrella (15.0%) IP: Poaceae (12.6%), Euphorbiaceae (6.6%), Fabaceae-Faboideae (4.2%), Heterofloral sample with primary contributions from Mimosa caesalpiniaefolia and Mimosa scabrella São Paulo-14 (Ribeirão Preto) PP: Mimosa caesalpiniaefolia (72.1%) IP: Poaceae (12.5%), Mi- mosa scabrella, (6.3%), Asteraceae (4.7%), Euphorbiaceae (4.4%) Heterofloral sample with a primary contribution from Mimosa caesalpiniaefolia São Paulo-15 (Ribeirão Preto) PP: Mimosa caesalpiniaefolia (64.5%) AP: Poaceae (19.3%) IP: Asteraceae (12.9%), Euphorbiaceae (3.2%) Heterofloral sample with primary contributions from Mimosa caesalpiniaefolia and Poaceae São Paulo-16 (Ribeirão Preto) PP: Schinus (58.6%) AP: Alternanthera (34.6%), Asteraceae (3.0%) Heterofloral sample with primary contributions from Schinus and Alternanthera São Paulo-17 (Ribeirão Preto) AP: Persea (25.9%), Myrcia (20.3%), Anadenanthera (18.6%), Eucalyptus (18.4%) IP: Cecropia (6.9%), Tapirira (4.7%), Asteraceae (3.5%) Heterofloral sample with primary contributions from Persea, Myrcia, Anadenanthera and Eucalyptus São Paulo-18 (Ribeirão Preto) PP: Schinus (82.3%) AP: Vernonia (15.3%) Heterofloral sample with primary contributions from Schinus and Vernonia São Paulo-19 (Ribeirão Preto) PP: Anadenanthera (82.5%) IP: Eucalyptus (10.0%), Cecropia (5.7%) Heterofloral sample with a primary contribution from Anadenanthera São Paulo-20 (Ribeirão Preto) AP: Cecropia (43.2%) Anadenanthera (33.9%) IP: Eucalyptus (12.3%), Myrcia (9.3%) Heterofloral sample with primary contributions from Cecropia and Anadenanthera São Paulo-21 (Taubaté) PP: Mimosa caesalpiniaefolia (46.1%) AP: Mimosa scabrella (37.2%) IP: Syagrus (10.2%) Heterofloral sample with primary contributions from Mimosa caesalpiniaefolia and Mimosa scabrella São Paulo-22 (Taubaté) PD: Mimosa scabrella (80.0%)IP: Malpighiaceae (5.9%) Heterofloral sample with a primary contribution from Mimosa scabrella São Paulo-23 (Taubaté) AP: Mimosa caesalpiniaefolia (43.1%), Antigonon leptopus (23.2%), Eucalyptus (15.4%) PI: Poaceae (11.0%), Cecropia (7.3%) Heterofloral sample with primary contributions from Mimosa caesalpiniaefolia, Antigonon leptopus and Eucalyptus São Paulo-24 (Taubaté) PP: Mimosa scabrella (92.3%) IP: Antigonon leptopus (4.9%) Monofloral sample of Mimosa scabrella PP = predominant pollen (> 45%); AP = accessory pollen (15-45%); IP = isolate pollen (3-15%) AS Freitas, VAS Arruda, LB Almeida-Muradian, OM Barth - Botanical profiles of pollen loads collected by Apis mellifera62 Table 3. The pollen types identified in the pollen load samples of A. mellifera collected in South and Midwest macro-region of Brazil and their main botanical origins, considering a frequency of more than 3%. (Samples were considered monofloral when presenting more than 90% of a unique pollen type). Municipality Brazilian macro-region = South Pollen analysis and pollen types Predominant botanical origin (representative pollen types) Santa Catarina-1 (Fraiburgo) PP: Asteraceae (54.2%); AP: Brassica (36.4%); IP: Amaranthus/Chenopodiaceae (3.3%); Euphorbiaceae (3.6%) Heterofloral sample with primary contributions from Asteraceae and Brassica Santa Catarina-2 (Fraiburgo) PP: Asteraceae (76.2%) AP: Brassica (20.0%) Heterofloral sample with primary contributions from Asteraceae and Brassica Santa Catarina-3 (Fraiburgo) PP: Asteraceae (56.3%) AP: Brassica (35.6%) IP: Euphorbiaceae (6.1%) Heterofloral sample with primary contributions from Asteraceae and Brassica Santa Catarina-4 (Fraiburgo) PP: Asteraceae (71.8%) AP: Brassica (19.7%) IP: Euphorbiaceae (7.0%) Heterofloral sample with primary contributions from Asteraceae and Brassica Santa Catarina-5 (Fraiburgo) PP: Asteraceae (56.7%) AP: Brassica (23.3%) IP: Euphorbiacae (10.0%), Amaranthus/ Chenopodiaceae (6.7%), Poaceae (3.3%) Heterofloral sample with primary contributions from Asteraceae and Brassica Santa Catarina-6 (Fraiburgo) PP: Asteraceae (50.0%) AP: Euphorbiaceae (26.6%), Brassica (18.7%) IP: Amaranthus/Chenopodiaceae (4.7%) Heterofloral sample with primary contributions from Asteraceae, Brassica and Euphorbiaceae Santa Catarina-7 (Campos Novos) PP: Caesalpiniaceae (63.1%) IP: Asteraceae (13.6%), Sebastiania (9.5%), Apiaceae (8.5%), Myrcia (3.8%) Heterofloral sample with a primary contribution from Caesalpiniaceae Santa Catarina-8 (Serra Catarinense) PP: Mimosa caesalpiniaefolia (85.2%) IP: Asteraceae (11.8%) Heterofloral sample with a primary contribution from Mimosa caesalpiniaefolia Rio Grande do Sul-1 (Santana do Livramento) PP: Ambrosia (99.4%) Monofloral sample of Ambrosia Rio Grande do Sul-2 (Santana do Livramento) PP: Eucalyptus (93.0%) IP: Asteraceae (7.0%); Monofloral sample of Eucalyptus Rio Grande do Sul-3 (Cruz Alta) PP: Brassica (57.4%) AP: Eucalyptus (42.6%) Heterofloral sample with primary contributions from Brassica and Eucalyptus Rio Grande do Sul-4 (Arvorezinha) PP: Eucalyptus (49.9%) AP: Eupatorium (24.5%) IP: Onagraceae (10.1%), Ilex (6.5%), Asteraceae (4.6%), Myrcia (4.1%) Heterofloral sample with primary contributions from Eucalyptus and Eupatorium Rio Grande do Sul-5 (São Gabriel) PP: Eucalyptus (64.6%) AP: Brassica napus (35.3%) Heterofloral sample with primary contributions from Eucalyptus and Brassica napus Distrito Federal PP: Schinus (92.1%) IP: Eucalyptus (4.8%), Cecropia (3.2%) Monofloral sample of Schinus Mato Grosso-1 (Sinop) PP: Cecropia (66.6%) AP: Euphorbiaceae (26.7%); IP: Poaceae (6.7%) Heterofloral sample with primary contributions from Cecropia and Euphorbiaceae Mato Grosso-2 (Sinop) PP: Cecropia (97.1%) Monofloral sample of Cecropia Mato Grosso-3 (Sinop) PP: Cecropia (51.3%) AP: Poaceae (44.3%) Heterofloral sample with primary contributions from Cecropia and Poaceae PP = predominant pollen (> 45%); AP = accessory pollen (15-45%); IP = isolate pollen (3-15%). 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