Pollen and physicochemical analysis of Apis and Tetragonisca () honey

Ortrud M. Barth, Alex S. Freitas, Graziela L. Sousa and Ligia B. Almeida-Muradian

SUMMARY

Pollen and physicochemical analyses of honey were car- rected to plants of the native flora. The assemblage of plant ried out in order to distinguish between trophic resources species visited during the flowering period by Apis and Tetrag- and nutritional preferences of Apis mellifera (honeybees) and onisca was not the same. Physicochemical analyses showed (‘ jataí’ bees). Honey samples from both differences between all analyzed parameters, except for color bee species were obtained on the same day in each apiary. determination. Jataí honeys presented higher values of humid- Six apiaries, localized in different regions of the State of São ity, acidity, diastase, ashes, sucrose and electric conductivity, Paulo, Brazil, contributed, in different days, to the experiment. and Apis honeys of fructose and glucose. No correspondence No bee species was more generalist than the other. Eucalyptus between physicochemical and monofloral honey properties was and Citrus, two introduced plant genera in Brazil, were very observed. attractive to honeybees. The preference of jataí bees was di-

Análisis fisicoquímico y del polen de mieles de Apis y Tetragonisca (Apidae) Ortrud M. Barth, Alex S. Freitas, Graziela L. Sousa y Ligia B. Almeida-Muradian RESUMEN

Se llevaron a cabo análisis fisicoquímicos y del polen de preferencia de las abejas jatai estuvo dirigida a plantas de la las mieles a fin de distinguir entre los recursos tróficos y las flora nativa. El conjunto de especies de plantas visitadas du- preferencias nutricionales de Apis mellifera (abejas de miel) rante el periodo de floración por Apis y Tetragonisca fue dife- y Tetragonisca angustula (abejas ‘ jataí’). Muestras de miel de rente. Los análisis fisicoquímicos mostraron diferencias entre ambas abejas fueron obtenidas el mismo día en cada apiario. todos los parámetros analizados, excepto el color. Las mieles Seis apiarios localizados en diferentes regiones del estado de jatai presentaron valores más altos de humedad, acidez, dias- Sao Paulo, Brasil, contribuyeron, en días diferentes, al estudio. tasa, cenizas, sacarosa y conductividad eléctrica, y las mieles Ninguna de las especies fue más generalista que la otra. Eu- de Apis valores de fructosa y glucosa. No se observó corres- calyptus y Citrus, dos géneros de plantas introducidas al Bra- pondencia entre las propiedades fisicoquímicas y la miel mo- sil, resultaron ser muy atractivas para las abejas de miel. La nofloral.

Introduction drastically reduced and food royal jelly (Barth, 2005; Mor- 2007) were applied to honey- competition increased between gado and Barth, 2011) were bee products only. Pollen spec- European bees (Apis sp.) bees, requiring investigation. later studied using pollen anal- tra of honeys were introduced into the The former melissopalyno- ysis as well. were presented by several au- southern region of Brazil since logical knowledge in Brazil Both palynological and thors (Vit and Ricciardelli 1839 by colonists (Wiese, was reviewed by Barth (2004). physicochemical analyses of D’Albore, 1994; Bazlen, 2000; 1984) intending commercial Trophic preferences of bees pollen loads (Bastos et al., Carvalho et al., 2001; Alves et activities. The native stingless and the phytogeographical 2004; Almeida-Muradian et al., 2006). Comparative physi- bees have had no value for characterization of Brazilian al., 2005) and honeys (Barth cochemical analyses between honey production. At present, honeys (Luz et al., 2007), geo- et al., 2005; Almeida-Muradi- propolis (honeybees) and geo- the natural bee flora has been propolis (Barth, 2006) and an et al., 2007; Luz et al., propolis (stingless bees) sam-

Keywords / Apis mellifera / Honey / Meliponinae / Physicochemical Analysis / Pollen Analysis / Tetragonisca angustula / Trophic Resources / Received: 05/04/2012. Modified: 04/01/2013. Accepted: 04/08/2013.

Ortrud Monika Barth. Ph.D. Rio de Janeiro, RJ, Brazil. e- Graziela L. Sousa. M.Sc. Re- Ligia Bicudo Almeida-Muradian. Researcher. Instituto Oswaldo mail: [email protected] searcher and Collaborator, De- Ph.D. Researcher and Collabo- Cruz (Fiocruz), Brazil. Address: Alex da Silva de Freitas. Biolo- partamento de Alimentos e rator, Departamento de Ali- Laboratório de Morfologia e gist. Collaborator, Universidade Nutrição Experimental, Facul- mentos e Nutrição Experimen- Morfogênese Viral, Pavilhão Federal Fluminense (UFF), dade de Ciências Farmacêuti- tal, Faculdade de Ciências Hélio e Peggy Pereira, Fiocruz. Niteroi, RJ, Brazil. cas, Universidade de São Paulo Farmacêuticas, USP, São Pau- Avenida Brasil 4365. 21040-900 (USP), São Paulo, SP, Brazil. lo, SP, Brazil.

280 0378-1844/13/04/280-06 $ 3.00/0 APR 2013, VOL. 38 Nº 04 AnálisE fisicoquímicA E dO pÓlen dO mel de Apis y Tetragonisca (Apidae) Ortrud M. Barth, Alex S. Freitas, Graziela L. Sousa e Ligia B. Almeida-Muradian RESUMO

Realizaram-se análises físico-químicas e do pólen dos méis A preferência das abelhas jataí esteve orientada a plantas da com o fim de distinguir entre os recursos tróficos e as prefe- flora nativa. O conjunto de espécies de plantas visitadas du- rências nutricionais de Apis mellifera (abelhas de mel) e Tetra- rante o período de floração por Apis e Tetragonisca foi dife- gonisca angustula (abelhas ‘ jataí amarela’). Mostras de mel de rente. As análises físico-químicas mostraram diferenças entre ambas as abelhas foram obtidas no mesmo dia em cada apiá- todos os parâmetros analisados, exceto a cor. Os méis jataí rio. Seis apiários localizados em diferentes regiões do estado apresentaram valores mais altos de umidade, acidez, diástase, de São Paulo, Brasil, contribuíram, em dias diferentes, ao es- cinzas, sacarosa e condutividade eléctrica, e os méis de Apis tudo. Nenhuma das espécies foi mais generalista que a outra. valores de frutose e glucose. Não se observou correspondência Eucalyptus e Citrus, dois gêneros de plantas introduzidas ao entre as propriedades físico-químicas e o mel monofloral. Brasil, resultaram ser muito atrativas para as abelhas de mel.

ples were presented by Pereira Table I stingless bees of Brazil, which et al. (2003). Localities where honey samples of Apis mellifera presents differences in some Trophic resources used by and Tetragonisca angustula were obtained physicochemical parameters honeybees and stingless bees (Azeredo et al., 2000). Samples / Global Physicochemical analyses were considered in relation to position system Localities / beekeepers Date of pollen loads also (Carvalho (GPS) collection were carried out in the Labo- and Marchini, 1999; Souza et ratory of Food Analyses of the Amparo Sítio Bela Vista / 25/08/2006 Department of Foods and Ex- al. 2002; Ramalho et al., 2007; 22o42'04''S Amparo-SP / Morgado et al., 2011). No 46o45'52''W Vicentin perimental Nutrition, Pharma- available palynological data is ceutical Science School, Uni- Itaberaba Apiário Capelinha / 16/04/2007 known for Brazilian honey o versity of São Paulo. The 23 27'46''S Estrada de Itaberaba, quality control of Apis mel- samples obtained in a same 46o31'58''W 7 Guarulhos-SP / day from both species of bees Julio de Oliveira lifera honeys moisture (%), living in a same apiary (desig- acidity (mEq·kg‑1), ashes (%) Lins Apacame Lins-SP / 20/03/2006 and diastase number followed nated ‘paired honeys’). When o honeybee and stingless bee 21 40'25''S Waldemar Monteiro the methodology of the AOAC 49o45'23''W combs were located closely in (1990), Brasil (1981) and CAC an apiary, competition of for- Marília Marília- BR 153 / 13/11/2007 (1989). Sugars were analyzed 49o56'46''S Cynthia Anderson de Souza by HPLC: glucose (%), fruc- aging activities must be stud- o ied. Considered sometimes as 23 13'10''W Carvalho Colombo tose (%) and sucrose (%) fol- specialists and sometimes as Pedreira Sítio Caxambu / 06/11/2006 lowing the methodology in generalists, these bees may 22o44'31''S Pedreira-SP / Bogdanov et al. (1997); the have some trophic resources in 46o54'05''W Luciano Panigassi electric conductivity in uS·cm‑1 common. Palynological and Sto. Antônio de Posse Apícola Del Fiore / 02/12/2007 was determined following the chemical analysis of these 22o36'22''S Marcelo Del Fiore methodology in Stefanini honeys (Vit et al., 2006) may 46o55'10''W (1984). The color determina- help to get a better knowledge tion (mm Pfund) followed the of the apiflora and, conse- methodology in Brasil (1981). quently, a better honey quality, and distant localities (Table I) 1978) to obtain the honey sed- Standard values were present- income and profit to beekeep- in the State of São Paulo, Bra- iments without the use of ac- ed (Marchini et al., 2004, ers. zil: Amparo, Itaberaba, Lins, etolysis. Available palynologi- 2005; Barth et al., 2005; Al- Therefore, we analyzed Marília, Pedreira and Santo cal literature (Barth, 1989: meida-Muradian and Bera, paired honey samples of Apis Antônio da Posse. The apiaries Roubik and Moreno, 1991; 2008; Souza, 2008). mellifera and Tetragonisca maintained combs of the hon- Moreti et al., 2002) and the angustula, and identified the eybee Apis mellifera (Apidae), pollen slide collection of the Results respective plants visited by and the stingless bee Tetrago- Laboratory of Palynology, Fed- these bees at the same time in nisca angustula (Meliponinae, eral University of Rio de Ja- The data obtained are pre- the same locality, using paly- ‘jataí’). The honey samples neiro, enabled pollen grain sented in Table II, comprising nological and physicochemical were obtained from both spe- identification. 20 pollen types of nectarifer- methods, in order to get infor- cies in each apiary and in the Standards of Apis mellifera ous plants, whose frequency mation about the competition same day by the beekeepers. honeys for commercialization is ≥3% of pollen grains count- for nectar resources, nutritional Six honey samples from A. used the Normative Instruction ed per sample (sum of grains). th 2000 (Bra- preferences and honey quality. mellifera (A) and six from T. 11, of October 20 Fourteen plant taxa were vis- angustula (T) were analyzed, sil 2000), based upon the Eu- ited by Apis mellifera and 18 ropean legislation. It only at- Material and Methods starting with 10g of well by Tetragonisca angustula. mixed honey, and using the tends the characteristics of the The percentage of unidentified The paired honey samples European palynological meth- honeybees and does not in- pollen grains remained <3%, proceeded from six distinct odology (Louveaux et al., clude the honey of native except in samples from Marí-

APR 2013, VOL. 38 Nº 04 281 Table II Relative frequency (%) OF NECTARIFEROUS pollen types (>3%) recognized in the honey samples in the localities of Amparo, Itaberaba, Lins, Marília, Pedreira and Santo Antônio da Posse, state of São Paulo Amparo Amparo Itaberaba Itaberaba Lins Lins Marília Marília Pedreira Pedreira Posse Posse Pollen types A T A T A T A T A T A T Amaranthaceae Alternanthera 4.5 Anacardiaceae 9.9 13.4 4.2 10.3 12.6 Schinus 24.7/AP 33.5/AP Arecaceae 4,3 4.0 4.6 27.7/AP 5.9 ∑ Asteraceae 33.3 12.1 5.3 7.2 Eupatorium 25.2/AP 55.6/PP Ichthyothere 30.7/AP Brassicaceae Brassica 4.6 3.9 8.3 Caesalpiniaceae Caesalpinia 7.1 Caricaceae Carica papaya 4.7 76.6/PP Fabaceae Crotalaria 11.7 Melilotus alba 15.5 64.8/PP Flacourtiaceae Casearia 8.4 Lauraceae Persea 6.7 3.6 Mimosaceae Leucena 14.7 Piptadenia 62.8/PP 3.9 Myrtaceae Eucalyptus 64.6/PP 29.3/AP 10.0 92.4/PP 23.7 12.5 11.3 Myrcia 22.2/AP 4.0 4.7 3.6 13.1 Rhamnaceae Hovenia/Scutia 10.5 9.5 3.7 Rubiaceae Coffea 5.6 Rutaceae Citrus 56.8/PP 26.5/AP 8.7 Not identified 1.4 2.0 3.0 0.5 0.2 0.6 9.7 6.4 1.0 1.6 9.6 11.4 Pollen sum * 365 405 233 1541 419 502 372 423 1255 557 208 437 Nectariferous pollen * 325 351 199 172 394 368 236 415 321 332 204 333

The number of nectariferous pollen grains (100%) was obtained excluding the anemophilous and polliniferous pollen grains from the pollen sum. A: Apis mellifera, T: Tetragonisca angustula, PP: predominant pollen, AP: accessory pollen, * number of counted pollen grains. lia and Santo Antônio da Pos- types of Arecaceae, Eucalyp- Physicochemical analyses 0.17 to 0.42%, diastase from se localities. tus, Myrcia and Schinus in (Table III) of Apis honeys 11.01 to 22.45DN, glycose Predominant pollen grains samples of Tetragonisca (Am- showed moisture variation from 20.92 to 25.95%, fruc- (PP, comprising more than paro-T, Lins-T), characterizing between 15.40 and 19.00%, tose from 29.77 to 33.57%, 45% of the nectariferous bifloral honeys when two of acidity from 16.82 to saccharose from 8.38 to pollen grains counted) be- these taxa occurred simulta- 32.47mEq·kg‑1, ashes from 24.00% and electric conduc- longed to Eucalyptus in two neously. Heterofloral honeys 0.11 to 0.26%, diastase from tivity from 466.00 to samples of Apis (Amparo-A contained in addition acces- 2.20 to 11.49DN, glycose 931.67uS·cm‑1. The honey col- and Lins-A) and one of Cit- sory pollen grains of Citrus, from 29.49 to 37.45%, fruc- or varied from white to light rus (Itaberaba-A). Samples Eupatorium and Ichthyothere tose from 41.52 to 47.53%, ambar. of Tetragonisca presented in several samples of Apis. saccharose from below the dominant pollen types of The isolated pollen taxa limit of detection to 2.68%, Discussion Carica papaya (Marília-T), (IP, comprising 3 to 15% of and electric conductivity from Eupatorium (Posse-T), Meli- the nectariferous pollen grains 216.67 to 557.00uS·cm‑1. The Samples from the Amparo re- lotus alba (Pedreira-T) and counted) standed out in addi- colors of honey samples var- gion. Five pollen types only Piptadenia (Itaberaba-T), tion to pollen types of Alter- ied from extra light to dark with frequencies over 3% of characterizing monofloral nanthera, Anacardiaceae, ambar. all counted nectariferous pol- honeys (Figure 1 a-f). Brassica, Caesalpinia, Case- Physicochemical analyses of len grains, represented the Accessory pollen types (AP, aria, Coffea, Crotalaria, Hov- Tetragonisca honeys showed nectariferous sources in the comprising 15 to 45% of the enia/Scutia, Leucena and Per- moistures between 23.40 and two honey samples. Apis bees nectariferous pollen grains sea in several samples of both 25.60, acidity from 21.65 to remained preferentially on Eu- counted) included pollen the bees. 63.85mEq·kg‑1, ashes from calyptus flowers, and visited in

282 APR 2013, VOL. 38 Nº 04 Figure 1. Predominant pollen types. a: Eupatorium (Asteraceae), b: Piptadenia (Mimosaceae), c: Carica papaya (Caricaceae). d: Eucalyptus sp. (Myrtaceae), e: Melilotus alba (Fabaceae), f: Citrus sp. (Rutaceae).

Nevertheless, four other pol- species was visited by these len types were found. The bees in common. sub-representation of orange Samples from the Lins region. pollen grains in honeys indi- Seven significant pollen types cates a higher nectar contribu- addition a wild Asteraceae tus pollen was abundant in all indicated plants visited by the tion of this plant than indi- species like Eupatorium. honey samples of Apis, but bees. Apis remained on Euca- cated by its pollen grains Tetragonisca used Eucalyptus occurred only in the Amparo lyptus flowers. Tetragonisca counted. The jataí bees did nectar also, but equally a sample of Tetragonisca honey. showed preferences mainly of not visited Citrus flowers, and Myrtaceae species like Myrcia, Schinus and palm flowers. No a palm tree and several spe- Samples from the Itaberaba its preference was of a Pip- trophic similarity between the cies of Asteraceae. Therefore, region. Plants represented by tadenia species, besides Schi- two bee species was observed. it seems that the nectar re- nine pollen types were visited nus (‘aroeira’), Brassica (‘mo- sources were not so abundant significantly. The preference starda’) and a Rhamnaceae Samples from the Marília re- for jataí preferences. Eucalyp- of Apis was for Citrus nectar. species. Therefore, no plant gion. Six pollen types were of

Table III Physicochemical analyses of honeys of Apis mellifera (A) and Tetragonisca angustula (T) in six localities of the state of São Paulo*

Chemical Amparo Amparo Itaberaba Itaberaba Lins Lins Marília Marília Pedreira Pedreira Posse Posse analyses A T A T A T A T A T A T Moisture 17.13 ±0.23 23.40 ±0.23 17.00 ±0.00 24.20 ±0.00 19.00 ±0.00 25.60 ±0.00 17.00 ±0.00 24.80 ±0.00 18.20 ±0.00 23.80 ±0.00 15.40 ±0.00 24.40 ±0.00 (%) Acidity 32.47 ±0.58 32.47 ±0.58 27.57 ±0.17 22.38 ±0.36 27.43 ±1.33 48.13 ±0.45 20.74 ±0.87 21.65 ±0.00 27.85 ±0.11 63.85 ±0.35 16.82 ±0.69 27.32 ±0.29 (mEq/kg) Ashes 0.26 ±0.02 0.25 ±0.00 0.11 ±0.01 0.20 ±0.00 0.25 ±0.01 0.17 ±0.01 0.22 ±0.01 0.40 ±0.02 0.18 ±0.00 0.42 ±0.02 0.16 ±0.00 0.23 ±0.01 (%) Diastase 9.66 ±0.20 17.19 ±1.56 2.20 ±0.11 22.45 ±0.68 4.40 ±0.19 18.10 ±0.82 11.49 ±0.46 11.01 ±0.21 7.01 ±0.02 18.69 ±0.30 9.16 ±0.29 14.15 ±0.04 (DN) Glycose 29.49 ±0.60 23.87 ±1.27 33.93 ±0.33 25.95 ±0.10 37.45 ±1.23 22.75 ±0.98 30.67 ±0.46 22.15 ±0.44 29.97 ±0.81 20.92 ±0.68 30.54 ±0.40 21.08 ±0.71 (%) Fructose 43.44 ±1.70 31.19 ±1.58 47.53 ±0.46 33.57 ±0.32 41.52 ±2.08 33.55 ±1.64 42.84 ±0.15 28.95 ±0.46 42.48 ±0.63 28.59 ±0.49 44.95 ±0.54 29.77 ±0.25 (%) Saccharose

* Each value represents the mean ±standard diversion of three analyses.

APR 2013, VOL. 38 Nº 04 283 interest to the bees, but two studied. When suffi- Table IV only were visited by both. cient flowering of Eu- Paired honey samples. Trophic resources used From these, Carica papaya calyptus occurred in by Apis mellifera (*) and Tetragonisca angustula (+) bees‡ was dominant for jataí only, Lins, Apis bees re- Apis Apis and Tetragonisca Tetragonisca while Apis visited Schinus mained on this tree, (exclusive) (superposition) (exclusive) and Eucalyptus flowers. Two while Tetragonisca vis- pollen types (C. papaya and ited mainly the native Myrcia) were found in both. flora of Schinus and a palm tree (like Sy- Samples from the Pedreira agrus, ‘gerivá’), show- region. Eleven pollen types ing clearly its trophic were significant in these honey Plant taxa types) (pollen Citrus Coffea Leucena Anacardiaceae Arecaceae Asteraceae Brassica Carica papaya Eucalyptus Myrcia Persea Rhamnaceae Alternanthera Brassica Caesalpinia Crotalaria Casearia Melilotus alba Piptadenia preference. samples. Nevertheless, Apis % Tetragonisca bees bees produced a heterofloral 100 showed an affinity to honey with a predominant par- species of the Faba- * ticipation of a wild Asteraceae; 90 ceae family as ob- Tetragonisca remained on Fa- served in samples of baceae flowers, like Melilotus 80 Lins and Pedreira. alba. Two pollen types (Schi- Similar behaviour of + nus and Eupatorium) were T. angustula was em- 70 found in both species. phazised by Carvalho * + + Samples from the Santo An- et al. (1999). 60 + tônio da Posse region. Seven Honey samples of * significant pollen types indi- Apis mellifera were 50 cated plants visited by the distinct in regard to bees, six of them by Apis, all physicochemical 40 mainly Citrus. This means parameters presented that flowering of orange when compared with * *+ 30 shrubs was not satisfactory honey samples of T. + + for honey production, and that angustula, except for * + * * wild plant species were visit- determination of its 20 + ed in addition. Tetragonisca color in mm Pfund. * *+ * * + 10 visited Citrus also, but the The physicochemi- *+ + * * + + + wild flowers of Eupatorium cal analyses showed * + *+ + + * + * + + 03 were of greater interest to the differences be- *+ + * + them. This observation dem- tween the honey sam- onstrates an inversion of the ples of Apis and ‡ Nectariferous plant taxa were identified using pollen analysis, and considered of 100% bee trophic preferences. Tetragonisca at each frequency. Taking all honey samples locality. The honeys into account, Apis produced of Apis contained always a diastase number, glucose and of the native flora was pre- monofloral honeys of Euca- higher concentration of glu- fructose. Honey of the Carica ferred by jataí bees. lyptus and Citrus in three lo- cose and fructose, while the papaya pollen type had the calities (Amparo, Itaberaba, honeys of Tetragonisca were lowest acidity value and that of 3) The assemblage of nectarif- Lins). Tetragonisca produced the richest of sucrose and pre- Melilotus alba pollen type pre- erous plant species visited monofloral honeys of Piptad- sented a high electric conduc- sented the highest values of during the flowering period enia, Carica papaya pollen tivity. The humidity, acidity ashes, sucrose and electric con- by Apis and Tetragonisca type, Fabaceae (Melilotus alba (except the Itaberaba sample), ductivity. No special data were bees was not the same. Lack pollen type) and Eupatorium ashes (except Amparo and recorded for Eupatorium honey. of food from native plant spe- in four localities of the state Lins samples) and diastase Considering all the data cies may probably induce the of São Paulo (Itaberaba, number (except Marilia sam- obtained using palynological jataí bees to visit introduced Marília, Pedreira and Santo ple) presented higher values and physicochemical analyses plant species. Antônio da Posse). in the Tetragonisca honeys. of the paired honey samples 4) Several pollen types could be Superposition at the same Considering the monofloral of A. mellifera and T. angus- determined at the specific, ge- time of important trophic re- honeys of Apis, the Citrus tula, the following observa- neric or family level. This dem- sources for Apis and Tetrago- honey sample showed the tions could be established: onstrated that our knowledge nisca bees (considering prepon- highest content of fructose 1) No bee species was more about the main nectar producing derant?? and accessory pollen and the lowest of ashes. The generalist than the other ac- apiflora needs more and better types) occurred only once in two monofloral Eucalyptus cording to the honey samples detailed local investigation. As a Amparo over Eucalyptus trees honeys showed the highest analyzed. Both bees produced rule, the vegetation around an (Table IV). This behavior may values of ashes and electric monofloral, bifloral and het- apiary in a radius of 1,5km be explained by deficient wild conductivity. erofloral honeys. must to be very well known. flower nectar resources for the In relation to the monofloral jataí bees, as it was observed honeys of Tetragonisca, that 2) Eucalyptus and Citrus, two 5) No correspondence between by Luz et al. (2007) for honey- from Piptadenia was the sole introduced plant taxa in Bra- the physicochemical properties bees also. Apis visited Eucalyp- white colored honey and pre- zil, were very attractive to and the monofloral pollen spec- tus flowers in all the localities sented the highest values of Apis bees, while plant species tra of honey samples of A. mel-

284 APR 2013, VOL. 38 Nº 04 lifera and T. angustula was ob- Barth OM (1989) O Pólen no Mel mel. Ministério da Agricultura, Papel Virtual. Rio de Janeiro, served. Brasileiro. Luxor. Rio de Janei- Pecuária e Abastecimento. Brazil. 93 pp. ro, Brazil. 150 pp. SisLegis - Sistema de Consulta à Pereira AS, Bicalho B, Aquino-Neto Legislação. Brazil http://extranet. ACKNOWLEDGEMENTS Barth OM (2004) Melissopalynology FR (2003) Comparison of propo- in Brazil: a review of pollen agricultura.gov.br/sislegisconsulta lis from Apis mellifera and analysis of honeys, propolis and (Cons. 02/05/2009). Tetragonisca angustula. Apidolo- The authors thank the Con- pollen loads of bees. Sci. Agric. CAC (1989) Codex Standards for gie 34: 291-298. Sugars (Honey). Codex Alimen- selho Nacional de Desenvolvim- 61: 342-350. Ramalho M, Silva MD, Carvalho tarius Commission. FAO. Rome, ento Científico e Tecnológico Barth OM (2005) Botanical resources CAL (2007) Dinâmica de uso de Italy.. (CNPq) and the Fundação de used by Apis mellifera deter- fontes de pólen por Melipona Amparo à Pesquisa do Estado mined by pollen analysis of Carvalho CAL, Marchini LC (1999) scutellaris Latreille (Hymenop- royal jelly in Minas Gerais, Bra- Tipos polínicos coletados por ters: Apidae): uma análise com- de São Paulo (FAPESP) for fi- zil. J. Apicult. Res. 44: 78-81. Nanotrigona testaceicornis e parativa com Apis mellifera L. nancial support and scholarship. Barth OM (2006) Palynological anal- Tetragonisca angustula (Hyme- (: Apidae), no Thanks are due to Adriana Mat- ysis of geopropolis samples ob- noptera, Apidae, Meliponinae). domínio tropical atlântico. Neo- suda from Centro Tecnológico tained from six species of Meli- Sci. Agric. 56: 717-722. trop. Entomol. 36: 38-45. de Análise de Alimentos (CE- poninae in the Campus of the Carvalho CAL, Moreti ACCC, Roubik DW, Moreno JE (1991) Pollen Marchini LC, Alves RMO, TAL) for sugars analysis. 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