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Anais da Academia Brasileira de Ciências (2010) 82(4): 893-902(Annals of the Brazilian Academy of Sciences)ISSN 0001-3765www.scielo.br/aabc

Evaluation of the botanical origin of commercial dry bee pollen load batchesusing pollen analysis: a proposal for technical standardization

ORTRUD M. BARTH1,2, ALEX S. FREITAS1, ÉRIKA S. OLIVEIRA1, ROSANA A. SILVA3,FERNANDA M. MAESTER3, RAQUEL R.S. ANDRELLA3 and GINA M.B.Q. CARDOZO3

1Laboratório de Palinologia, Departamento de Botânica, Instituto de Biologia, Universidade Federal do Rio de Janeiro,Ilha do Fundão, Centro de Ciências da Saúde, Bloco A, Sala 118, 21949-900 Rio de Janeiro, RJ, Brasil

2Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Avenida Brasil 4365, 21040-900 Rio de Janeiro, RJ, Brasil3ITAL – Instituto de Tecnologia de Alimentos, Centro de Ciência e Qualidade de Alimentos,

Avenida Brasil 2880, 13070-178 Campinas, SP, Brasil

Manuscript received on August 20, 2009; accepted for publication on April 8, 2010

ABSTRACT

High quality of bee pollen for commercial purpose is required. In order to attend the consumer with the best identifi-cation of the botanical and floral origin of the product, 25 bee pollen batches were investigated using two techniquesof pollen grain preparation. The first started to identify pollen loads of different colors in two grams of each well mixedbatch, and the second to identify pollen grains in a pool made of all the pollen loads comprised in two grams. The bestresult was obtained by this last technique, when a pollen grain suspension was dropped on a microscope slide and circa500 pollen grains were counted per sample. This analysis resulted in the recognition of monofloral and bifloral pollenbatches, while the use of the first technique resulted in all samples receiving a heterofloral diagnosis.

Key words: Apis, pollen loads, commercial pollen batches, pollen analysis, botanical origin.

INTRODUCTION

Bee pollen production increased during the last yearsas a response to commercial demand. Apis mellifera L.is the best pollen supplier mainly in tropical countrieswhere entomophilous plant species are dominant. Bee-keepers use pollen traps of different types in order toobtain the pollen loads from bees when coming hometo hives. This pollen is humid and has to be dried be-fore commercialization. Then, it may be distributed invials receiving an identification that comprises, in addi-tion, its botanical origin. This procedure is importantto avoid missing the accurate scientific definition of thebotanical name, since beekeepers report a lot of commonnames of plants that were visited by the bees.

Pollen grains present a huge variation of morpho-logical features that are established by its genetic hered-

Correspondence to: Ortrud Monika BarthE-mail: barth@ioc.fiocruz.br

ity and are not influenced by environmental events orchanges. Pollen analysis is the tool to recognize fromwhere the pollen grains are coming. Several laborato-rial techniques are used to prepare pollen grains for mi-croscope observation and identification. The difficultyremains in the interpretation of the obtained data andthe evaluation of numerical and personal informations.Several aspects are to be considered, the first one regard-ing the bee behavior upon flower visitation for pollenextraction. In this case, single bees are captured andboth of their pollen loads were analyzed (Carvalho andMarchini 1999, Marques-Souza et al. 2002, Noor et al.2009). Secondly, when pollen traps are used, severalpollen loads perform one sample that, for analysis, hasto be well mixed; a group of pollen loads, in general se-lected by its weight or color (Almeida-Muradian 2005,Barth et al. 2009, Modro et al. 2009a), is used for therecognition of the botanical species composing thispollen sample.

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894 ORTRUD M. BARTH et al.

The main issue to address is the significance ornot of the choice by color. An accurate description ofthe pollen load sample processing and analysis is fre-quently missed during experiment descriptions (Modroet al. 2007, 2009a). Commonly, a fragment of a pollenload of each color is analyzed; no sample centrifuga-tion is used. In another way (denoted technique 1), tenpollen loads (Barth et al. 2009) or 25 ones of each colorbatch (Barth, personal information and in the presentpaper) are mixed using ethanol, and a little amount isanalyzed after centrifugation. As pollen grains have dif-ferent weight, a gradient is obtained, from which pollengrains are removed using a piece of glycerin jelly. Thesignificance of the obtained results is not clear. Anothertechnically very complex preparation of pollen batchesidentification was recently used comprising several stepsof pollen drying, hydrating and acetolysis (Novais et al.2009) and, thus, missing important information besidespollen morphology.

The proposed technique by this study (technique 2)may help to get quick and accurate information about thebotanical and regional origin of commercial bee pollenload products. It neither demands long time to be per-formed nor is expensive.

MATERIALS AND METHODS

Twenty five samples of commercial batches of bee pol-len loads were obtained in apiaries of several states inBrazil and analyzed (Table I). Each sample was analyzedusing two techniques. The first technique considers subsamples according to the pollen load colors; each samplewas made of well-mixed two grams and each sub sam-ple was made of 25 units or pollen loads of each color.When two or more types of color were mentioned in asame sub sample (Table I, column 2), it indicates thatno sufficient pollen loads of a unique color were avail-able to perform n=25, and several poorly representedcolor batches were mixed. Color was determined ac-cording to the Red Green Blue (RGB) color classifica-tion system. The color values correspond to: bright =RGB 247/240/183; brown = RGB 186/97/97, caramel =RGB 237/202/29, dark = RGB 117/11/11, green = RGB98/156/128, orange = RGB 255/106/37, violet = RGB123/102/147, yellow = RGB 250/224/23. The secondtechnique used all pollen loads together comprised intwo grams of a commercial batch sample.

Pollen grain identification used the available litera-ture, meanly Barth (1989), Roubik and Moreno (1991),and the Palynological Slide Collection of the Labora-tory of Palynology. Pollen frequency standard classes(Louveaux et al. 1978) were used, making an additionalsubdivision of the dominant pollen class (Table I).

TECHNIQUE 1 (TABLE I, COLUMNS 2-4):

Preparation of pollen load samples:

The pollen loads of two grams of each sample were dis-tributed in batches or sub samples according to its color.

Preparation of microscope slides:

Twenty five pollen loads of each sub sample were mac-erated using 10 mL of 70% ethanol distributed intotwo 15 mL centrifuge tubes and centrifuged during 3minutes at 1500 rpm. The sediment was resuspendedwith 10 mL of 70% ethanol and centrifuged again. Insequence, 5 mL of a water/glycerin 1:1 mixture wereadded to each tube, stirred, and left for 30 minutes. Af-ter centrifugation, the tubes remained with their aper-ture down on absorbent paper during a few minutes.Then, the pollen sediment was mixed and pollen slideswere prepared using a little piece of glycerin jelly tocapture the pollen grains, and slides were sealed withparaffin. Counting and identification comprised at least500 pollen grains.

TECHNIQUE 2 (TABLE I, COLUMNS 5-6):

Preparation of pollen load samples:

Dry pollen loads of two grams of a sample (this is a poolof circa 300 pollen loads) were weighted into a 15 mLFalcon centrifuge tube, mixed using 70% ethanol justto complete 13 mL, and left for 30 minutes. Treatmentwith ultrasound (if disposable) during 5 minutes can besuitable for particle dispersal. The sediment obtainedafter centrifugation may be extracted with ethanol, andsubmitted to ultrasonic treatment again. A solution ofdistilled water/glycerin 1:1 was added to the sedimentjust to complete 13 mL, and left for circa 30 minutes.

Preparation of microscope slides:

One drop of the well-mixed pollen grain suspension wasapplied on a microscope slide, covered with a 22×22 mmcover slide, and sealed with enamel. The slide may be

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STANDARDIZATION OF DRY BEE POLLEN ORIGIN EVALUATION 895

Fig. 1 – A pool of pollen loads prepared using technique 2; pollen grains are well dispersed all over the slide. Fig. 2 – A pool of pollen loads

prepared using technique 2 three weeks later; the slide started to dry, and the pollen grains overlapped and were pressed against the air bubbles.

maintained in a horizontal position for circa one monthor less, when drying starts, and the pollen grains wereselectively pressed against the air bubbles. Then discardthe slide (Figs. 1 and 2).

Preparation of stock samples:

One mL of the well mixed pollen grain suspension wasput into a 1.5 mL Eppendorf centrifuge tube and cen-trifuged. A 15 mL Falcon tube can be used as a supportfor the Eppendorf tube. After discarding the supernatant,one mL of glycerin was added, mixed, and then the tubewas closed and identified for additional preparations. Itmay be kept at room temperature.

Preparation of additional pollen slides:

The content of the Eppendorf tube has to be homoge-nized. One drop of this pollen grain suspension wasmixed with one drop of distilled water in a clean Ep-pendorf tube. A microscope slide, as formerly describedmay be prepared after 30 min waiting for water impreg-nation.

Counting and identification of pollen grainsand structured elements:

At least 500 pollen grains have to be counted and iden-tified using a 400× magnification, preferentially within10 days, before drying starts.

RESULTS AND DISCUSSION

The obtained data are shown in Table I. Each of the25 samples analyzed presented two or more colors of

pollen loads, establishing several sub samples. No sam-ple was single colored. When no sufficient pollen loadsof a unique color were available, two, rarely three dif-ferent colored pollen loads were joined together in orderto make up a group of 25 unities (column three); thesenever constituted a dominant sub group in a sample orbatch.

Fifty two pollen types presenting a frequency equalor higher than 3% of the pollen sum were recognized.Technique 1 presented 46 pollen types, 22 being exclu-sives. Technique 2 presented 29 pollen types, six beingexclusives.

No monofloral pollen load batch was recognizedby technique 1. Among the 25 analyzed samples, threewere bifloral presenting two dominant pollen types; 22samples were heterofloral comprising three or more pol-len types. Considering technique 2, eight pollen loadbatches were monofloral, six bifloral and 11 heterofloral.

Nevertheless, technique 1 detected more pollentypes (46) than technique 2 (29), this last procedureshowed a better evaluation of the whole pollen batch.Thus, commercial qualification has to follow this way.Consequently, it presented more monofloral batches.

Technique 1 was useful in order to teach about pol-len loads botanical origin. As explained below, the colorof pollen of a plant species may vary (Barth et al. 2009and in the present paper). Therefore, it is not a goodcharacteristic to inform about the botanical origin of apollen load batch. While it is not possible to analyze thepollen grain composition of each pollen load, and sev-eral plants may contribute to form a unique load, tech-nique 2 is a quicker and more accurate procedure tomake a diagnosis of a large commercial product.

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TABLE IComparison of 25 pollen load samples (2g of each batch) from Apis mellifera evaluated by its color

(sub samples in columns two, three and four) and evaluated by a pool of 2g of each batch (columns five and six).Each sub sample comprised 25 units of pollen loads. Only the pollen types with frequency >3% were considered:

very frequent (++++, circa >85%), frequent (+++, circa 45 to 85%), few frequent (++, circa 15 to 45%), rare (+, circa 3 to 15%).

Technique 1 Technique 2Sample Sub samples Pollen types Evaluation of Pollen types Evaluation ofregister and established identified in the the sub samples identified in a pool a pool of 2gprocedence by color of sub samples of 2g of the sample of the sample

pollen loads

55-Piaui yellow Cyperus (++++) monofloral of Cocos nucifera (+++) bifloral ofCyperus Cyperus (++) Cocos nucifera

bright and Cocos nucifera monofloral of Amaranthaceae (+) (dominant)green (++++) Cocos nucifera Solanaceae (+) and Cyperusdark Tapirira (++++), monofloral of

Bombax (+) Tapirirabrown Inga (++++), monofloral of

Cocos nucifera (++) Inga56-Bahia bright and Syagrus (++++), monofloral of Cocos nucifera monofloral of

orange Portulaca (++) Syagrus (++++) Cocos nuciferabright and Mimosa scabrella monofloral ofgreen (++++), Fabaceae (++) Mimosa scabrella

57-Bahia bright and Syagrus (++++), monofloral of Mimosa scabrela monofloral oforange Portulaca (++) Syagrus (++++) Mimosabrown and several polen heterofloral Syagrus (+) scabrellagreen types Cocos nucifera (+)

58-Minas Gerais bright Brassica (+++), monofloral of Eucalyptus (+++) heterofloralVernonia (+) Brassica Cecropia (++) with dominance

orange Senecio (++++) monofloral of Myrcia (+) of EucalyptusSenecio Syagrus (+)

dark Vernonia (++++) monofloral ofand uredospores Vernonia and

uredospores59-São Paulo caramel Poaceae (+++) monofloral of Eucalyptus (+++) heterofloral

Anadenanthera (+) Poaceae Vernonia (++)bright Syagrus (++++) monofloral of Senecio (++)

Syagrus Piper (+)dark and Eucalyptus (++++) monofloral of Myrcia (+)yellow Eucalyptusorange without pollen without pollen

grains, only grains, onlyuredospores uredospores

65-Bahia yellow and Cyperus (++++) monofloral of Mimosa scabrella monofloral ofbrown Cyperus (++++) Mimosabright Astrocaryum (++), heterofloral scabrella

Mimosa scabrella (++),Asteraceae (++)

dark immature pollen heterofloralgrains, uredosporesand other pollen types

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STANDARDIZATION OF DRY BEE POLLEN ORIGIN EVALUATION 897

TABLE I (continuation)

Technique 1 Technique 2Sample Sub samples Pollen types Evaluation of Pollen types Evaluation ofregister and established identified in the the sub samples identified in a pool a pool of 2gprocedence by color of sub samples of 2g of the sample of the sample

pollen loads

65-Bahia green Borreria monofloral ofdensiflora (++++) Borreria

densiflora

72-Espírito Santo bright and Euterpe edulis (+++), bifloral of Vernonia (++) heterofloralbrown Vernonia (++) Euterpe edulis Cecropia (++)

and Vernonia Euterpe edulis (+)orange and Persea (+++) heterofloral with Myrcia (+)yellow and several dominance of Eupatorium (+)

pollen types Persea uredospores (+)red and without pollen without pollenviolet grains, only grains, only

uredospres uredospores

74-Bahia yellow Syagrus (+++) without pollen Cecropia (++) heterofloralgrains, only Caesalpiniauredospores peltophoroides (++)

bright Cocos nucifera (+++), bifloral of Cocos Asteraceae (++)Astrocaryum nucifera and Cyperus (+)aculeatissimum Astrocaryum Syagrus (+)(+++) aculeatissimum Cocos nucifera (+)

dark Mangifera indica without pollen Myrcia (+)(++++) grains, only

uredosporesMangifera indica

orange and Borreria bifloral of Borreriagreen verticillata (+++), verticillata and

Bidens (++) Bidensbrown and Cocos nucifera (+++), bifloral of Cocosorange Cyperus (+++) nucifera and Cyperus

77-Sergipe bright and Cocos (++++), monofloral of Mimosa scabrella bifloral oforange Montanoa (+) Cocos (+++) Mimosabrown Cocos nucifera bifloral of Cocos Cocos nucifera (++) scabrella

(+++), Mimosa nucifera and Eupatorium (+) (dominant)caesalpiniaefolia Mimosa and Cocos(+++) caesalpiniaefolia nucifera

80-Sergipe bright Cocos nucifera monofloral of Mimosa scabrella bifloral of(++++), Cocos nucifera (+++) MimosaRichardia (+) Cocos nucifera (++) scabrella

dark and several pollen heterofloral Myrcia (+) (dominant)caramel types and Cocosorange Caesalpinia monofloral of nucifera

peltophoroides Caesalpinia(++++) peltophoroides

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TABLE I (continuation)

Technique 1 Technique 2Sample Sub samples Pollen types Evaluation of Pollen types Evaluation ofregister and established identified in the the sub samples identified in a pool a pool of 2gprocedence by color of sub samples of 2g of the sample of the sample

pollen loads

83-São Paulo bright, green Schinus (+++), bifloral of Cecropia (+++) monofloral ofand brown Cecropia (+++) Schinus and Senecio (+) Cecropia

and others Cecropia Trema (+)orange Senecio (++++) monofloral of Myrcia (+)

Senecio Eupatorium (+)Vernonia (+)

84-Minas Gerais yellow Antigonon monofloral of Antigonon leptopus monofloral ofleptopus (++++) Antigonon (+++) Antigonon

leptopus Baccharis (++) leptopusbright Baccharis (+++) monofloral of Mimosa scabrella

and others Baccharis (+), Mimosaorange and several pollen heterofloral caesalpiniaefoliabrown types (+), unknown (+)

85-Minas Gerais yellow Antigonon monofloral of Cecropia (+++) bifloral ofleptopus (+++), Antigonon Myrcia (++) CecropiaCyperus (+) leptopus Vernonia (+) (dominant)

bright and Anadenanthera (++), heterofloral and Myrciabrown Asteraceae (++)dark Myrcia (++++) monofloral of

Myrcia86-São Paulo yellow Eucalyptus (++++) monofloral of Eucalyptus (+++) monofloral of

Eucalyptus Eucalyptusorange Eucalyptus (+++), bifloral of

Euphorbiaceae (+++) Eucalyptus andEuphorbiaceae

brown Eucalyptus (++++) monofloral ofEucalyptus

87-Minas Gerais bright and Eucalyptus (++), bifloral of Eucalyptus (+++) monofloral ofbrown Vernonia (++), Eucalyptus and Vernonia (+) Eucalyptus

Croton (+) Vernoniaviolet, orange Eucalyptus (+++), bifloral ofand yellow Senecio (+++) Eucalyptus and

Senecio88-São Paulo yellow Baccharis (++++) monofloral of Eucalyptus (+++) monofloral of

Baccharis Poaceae (+) Eucalyptusbright Baccharis (++), bifloral of

Syagrus (++) Baccharis andSyagrus

brown Eucalyptus (++++) monofloral ofEucalyptus

97-Santa Catarina yellow Ilex (++++), monofloral of Eupatorium (++), heterofloralOnagraceae (+) Ilex Melastomataceae

bright Asteraceae (+++) monofloral of (++), Eucalyptusand others Asteraceae (++), Vernonia (+),

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TABLE I (continuation)

Technique 1 Technique 2Sample Sub samples Pollen types Evaluation of Pollen types Evaluation ofregister and established identified in the the sub samples identified in a pool a pool of 2gprocedence by color of sub samples of 2g of the sample of the sample

pollen loads

97-Santa Catarina orange Senecio (++++) monofloral of Montanoa (+)Senecio

brown Vernonia (++++) monofloral ofVernonia

100-Paraná yellow Alchornea (+++) monofloral of Sebastiania (+++), heterofloralAlchornea Brassica (++),

bright several pollen heterofloral Eucalyptus (++)types

orange and several pollen heterofloralbrown typesgreen Rosaceae (++++) monofloral of

fruits (plum,apple, pear)

red and Eucalyptus monofloral ofbrown (++++) and Eucalyptus and

uredosporos uredospores111-Santa Catarina bright Eupatorium (+++) bifloral of Vernonia (++) heterofloral

Cocos nucifera (++) Eupatorium and Myrcia (++)Vernonia (+) Cocos nucifera Montanoa (++)

orange Montanoa (++++) monofloral of Syagrus (+)Montanoa Poaceae (+)

brown Vernonia (++++) monofloral of Rubiaceae (+)Vernonia Crotalaria (+)

red Baccharis (+++) trifloralSebastiania (++)Montanoa (++)

115-Sergipe bright Cocos nucifera monofloral of Mimosa scabrella bifloral of(++++) Cocos nucifera (+++), Cocos Mimosa

orange Mimosa scabrella monofloral of nucifera (++) scabrella(+++), Asteraceae Mimosa (dominant)(several pollen scabrella and Cocostypes) nucifera

brown Mimosa scabrella monofloral of(++++), Cocos Mimosanucifera (++), scabrellaCommelina (++)

116-Sergipe yellow Syagrus (++++) monofloral of Mimosa bifloral ofSyagrus caesalpiniaefolia Mimosa

bright Cocos nucifera monofloral of (+++), Cocos caesalpiniaefolia(++++) Cocos nucifera nucifera (++) and Cocos

orange several pollen heterofloral nuciferatypes (dominant)

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900 ORTRUD M. BARTH et al.

TABLE I (continuation)

Technique 1 Technique 2Sample Sub samples Pollen types Evaluation of Pollen types Evaluation ofregister and established identified in the the sub samples identified in a pool a pool of 2gprocedence by color of sub samples of 2g of the sample of the sample

pollen loads

116-Sergipe brown Mimosa monofloral ofcaesalpiniaefolia Mimosa(++++), caesalpiniaefoliaMimosaceae Mv (++)

144-Bahia yellow Syagrus (+++), monofloral of Baccharis (+++), heterofloralCocos nucifera (+), Syagrus Cocos nucifera (++)Eupatorium (+) Syagrus (+)

bright Cocos nucifera monofloral of(++++) Cocos nucifera

several colors several pollen heterofloraltypes

145-Bahia caramel Cocos nucifera bifloral of Cocos Mimosa scabrella heterofloral(+++), Poaceae nucifera and (++), Schinus (++),(+++) Poaceae Poaceae (+),

bright Cocos nucifera bifloral of Cocos Cocos nucifera (+),(+++), nucifera and MimosaAstrocaryum Astrocaryum caesalpiniaefoliaaculeatissimum aculeatissimum (+), Eupatorium (+)(+++) Syagrus (+)

several colors Mimosa heterofloralcaesalpiniaefolia(++), Trema (++),and others

146-Bahia caramel Poaceae (++++) monofloral of Cocos nucifera (++) heterofloralTriumfetta (+) Poaceae Schinus (++)

bright Cocos nucifera monofloral of Poaceae (++)(++++) Cocos Mimosa

dark and several pollen heterofloral caesalpiniaefolia (++)yellow types Mimosa scabrella (+)orange Portulaca (+++) bifloral of

Commelina (++) Portulaca andCommelina

150-Espírito Santo yellow Alchornea (+++), monofloral of Eucalyptus (++) heterofloralArecaceae (+) Alchornea Cecropia (++)and others Myrcia (++)

caramel Poaceae (+++), monofloral of Asteraceae (+)Croton (+) Poaceae Cocos nucifera (+)

bright and Cocos nucifera bifloral of Cocos Poaceae (+)brown (+++), Syagrus nucifera and Alchornea (+)dark and red several pollen heterofloral

typesorange and Montanoa monofloral ofgreen (++++), Montanoa

Asteraceae (+)

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STANDARDIZATION OF DRY BEE POLLEN ORIGIN EVALUATION 901

Preparing slides in five folds, the same pollen spec-trum was obtained.

Detailed discussion of the data obtained by the useof the two techniques of pollen load batches analysis(Table I).

Pollen grain color of a plant taxon (third columnin Table I) can change from bright to dark as it maybe observed in Cocos (samples 55, 74, 77, 115, 145,146), Vernonia (samples 58, 111), Eucalyptus (samples86, 87) and Baccharis (sample 88) pollen grains, and inthe Mimosa scabrella pollen type (sample 115). Timeto air exposition of pollen grains resulting in exine andcytoplasm oxidation could be responsible for differentcolored pollen loads of a plant species.

Using technique 1, all pollen batches were hetero-floral. Technique 2 revealed monofloral (samples 56, 57,65, 83, 84, 86, 87, 88) and bifloral (samples 55, 77, 80,85, 115, 116) samples besides the heterofloral ones.

The monofloral samples presented five dominantpollen types: Cocos nucifera and Mimosa scabrella(= M. sensitiva) pollen types from the state of Bahia,Cecropia and Eucalyptus from São Paulo State, Antigo-non leptopus (a garden species) and Eucalyptus from thestate of Minas Gerais. These commercial batches haveobtained the best evaluation.

The bifloral samples presented six important pol-len types: Cocos nucifera and Cyperus from the stateof Piaui, Mimosa scabrella and Cocos nucifera in threesamples from the state of Sergipe, Mimosa caesalpiniae-folia and Cocos nucifera also from the state of Sergi-pe, and Cecropia and Myrcia from the state of MinasGerais. These commercial batches may receive a goodevaluation.

The heterofloral pollen load batches analyzed bytechniques 1 and 2 do not have palynological definitionof any dominance. Their botanical origin is variable de-pending upon several factors and their reproduction mustnot be effective.

A curious composition of some pollen loads wasmade of uredospores of fungi detected by technique 1 (insamples 59, 72, 74, 100). Uredospores never appearedusing technique 2. As so, their contribution to any ofthe pollen batches was not significant. Cladosporiumsp. spores were also collected by bees in the state ofMinas Gerais during an alimentary scarcity (Modro etal. 2009b).

In conclusion, using technique 1 based upon coloranalysis, more pollen types were identified, but no dom-inance of pollen type or plant species in a commercialpollen batch was reported. When using the technique2, besides the monofloral batches, more bifloral and lessheterofloral batches were recognized. This result showsthat a better characterization of a large pollen load batchcomposition of commercial interest was obtained whenusing the last technique.

RESUMO

É exigida alta qualidade para a comercialização de pólen

apícola. A fim de atender o consumidor com a melhor iden-

tificação da origem botânica e floral do produto, 25 partidas

de pólen apícola foram investigadas usando duas diferentes

técnicas na preparação dos grãos de pólen. A primeira par-

tiu da identificação das cargas polínicas contidas em dois gra-

mas de cada partida bem misturada segundo suas cores. A se-

gunda visava identificar os grãos de pólen de um agrupamento

(“pool”) de todas as cargas polínicas contidas em dois gramas

de cada amostra. O melhor resultado foi obtido pela última

técnica, quando uma suspensão de grãos de pólen era gote-

jada sobre uma lâmina de microscopia e cerca de 500 grãos de

pólen eram contados por amostra. Esta análise resultou no re-

conhecimento de partidas monoflorais e biflorais de pólen apí-

cola, enquanto que usando a primeira técnica, todas as amostras

receberam a diagnose heterofloral.

Palavras-chave: Apis, cargas de pólen, partidas comerciais de

pólen, análise polínica, origem botânica.

REFERENCES

ALMEIDA-MURADIAN LB, PAMPLONA LC, COIMBRA SAND BARTH OM. 2005. Chemical composition and bo-tanical evaluation of dried bee pollen pellets. J Food com-pos anal 18: 105–111.

BARTH OM. 1989. O pólen no mel brasileiro. Editora Luxor,151 p.

BARTH OM, MUNHOZ MC AND LUZ CFP. 2009. Botan-ical origin of Apis pollen loads using color, weight andpollen morphology data. Acta aliment 38: 133-139. DOI:10.1556/AAlim.2008.0026.

CARVALHO CAL AND MARCHINI LC. 1999. Tipos polínicoscoletados por Nannotrigona testaceicornis e Tetragoniscaangustula (Hymenoptera, Apidae, Meliponinae). Sci Agri(Piracicaba, Braz.) 56: 717–722.

An Acad Bras Cienc (2010) 82 (4)

“main” — 2010/11/4 — 14:29 — page 902 — #10

902 ORTRUD M. BARTH et al.

LOUVEAUX J, MAURIZIO A AND VORWOHL G. 1978.Methods of melissopalynology. Bee World 59: 139–157.

MARQUES-SOUZA AC, MIRANDA IPA, MOURA CO, RA-BELO A AND BARBOSA EM. 2002. Características mor-fológicas e bioquímicas do pólen coletado por cinco espé-cies de Meliponíneos da Amazônia Central. Acta Amaz32: 217–229.

MODRO AFH, MESSAGE D, LUZ CFP AND MEIRA-NETO

JAA. 2007. Composição e qualidade de pólen apícola co-letado em Minas Gerais. Pesqui agropecu bras 42: 1057–1065.

MODRO AFH, SILVA IC, LUZ CFP AND MESSAGE D.2009a. Analysis of pollen load based on color, physico-chemical composition and botanical source. An Acad BrasCienc 81: 281–285.

MODRO AFH, SILVA IC, MESSAGE D AND LUZ CFP.2009b. Saprophytic fungus collection by Africanized beesin Brazil. Neotrop Entomol 38(3): 434–436.

NOOR MJ, KAHN MA AND CAMPHOR ES. 2009. Paly-nological analysis of pollen loads from pollen sources ofhoneybees in Islamabad, Pakistan. Pak J Bot 41: 495–501.

NOVAIS JS, LIMA LCL AND SANTOS FAR. 2009. Botanicalaffinity of pollen harvested by Apis mellifera L. in a semi-arid area from Bahia, Brazil. Grana 48: 224–234.

ROUBIK DW AND MORENO JE. 1991. Pollen and sporesof Barro Colorado Island. Missouri Botanical Garden,Monographs in Systematic Botany, 263 p.

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