Propolis is a product elaborated by honey bees ( Apis ) and comprises plant resins, bee wax, glandular secretions of the worker bees and pollen grains. Geopropolis is a product elaborated by stingless bees and comprises similar compounds of honey bees and soil, mud, clay, earth or sand. The present study intends to distinguish between propolis and geopropolis using pollen analysis. A total of 12 samples were obtained in the Southern Brazilian macro-region and processed by standard melissopalynological methodology. All structural components recovered after alcoholic extraction and before acetolysis treatment were evaluated. Four of the eight samples collected in the State of Paraná were prepared by Apis and showed a strong contribution of Asteraceae and Eucalyptus pollen grains. On the other hand, the two geopropolis samples of Tetragonisca angustula contained mainly Cecropia pollen grains, while the samples of Melipona quadrifasciata and Melipona mondury showed a predominance of Melastomataceae pollen grains. The four propolis samples obtained in the Santa Catarina and Rio Grande do Sul States were characterized by a predominant pollen contribution of Asteraceae and Eucalyptus . Additional structured elements in propolis samples comprised frequently plant trichomes and tissue fragments. On the contrary, the geopropolis samples showed high values of spores of fungi, amorphous organic matter, sand or clay and sometimes remainder of resin. In conclusion, the pollen grain spectra do not distinguish between propolis of Apis and geopropolis of stingless bees, but the accessory elements are conclusive.
Honey bees (Apis sp.) were introduced in Southern Brazil by European immigrants, mainly Jesuit priests, in the XVII century, into the region of Missões, banks of the River Uruguai [
At this time, an African bee species (Apis mellifera adansonii Latreille, 1804) was accidentally introduced in the Brazilian state of São Paulo, Ribeirão Preto locality. The hybrid bees originated turned strongly aggressive and dominant and spread from this county into Southern Brazil too [
Propolis is a product elaborated by honey bees starting to collect plant resins into its basquetes. This resin was obtained from some damaged tree stems or branches [
Physico-chemical analyses of propolis revealed the presence of several substances such as flavonoids, ter- penoids and amino acids, antioxidant activity and more [
Structural elements in propolis samples were isolated as a rule by ethanol extraction. After elimination of resins and bee-wax, pollen grains (more or less 5% of the sediment amount), hyphae and spores of fungi, yeast, bacteria, plant tissue fragments and trichomes and organic clusters can be recovered [
Numerous species of native stingless bees, the Meliponini, are important pollinators of the original vegetation. In the same way as honeybees do, these bees elaborate a compound similar of propolis, named geopropolis. This term was formerly purposed by Nogueira-Neto [
Nests of Meliponini occur originally inside tree holes, inside walls and between stones [
In attending scientific and commercial interests, analysis of pollen grains and other structured elements that occur inside propolis and geopropolis is useful. It is possible to identify the phytogeographical region [
Physico-chemical analyses revealed geopropolis has a similar composition as propolis. The main difference is a variable concentration of each significant component. Phenolic acids and flavonoids mainly present lower concentration in geopropolis [
The Brazilian flora belongs to five macro-regions (North, Northeast, Midwest, Southeast and South). The Southern region (
The ombrophilous and the Araucaria angustifolia forests characterize the vegetation of Santa Catarina State [
The present study intends to distinguish between propolis and geopropolis by using a palynological analysis. It also intends to recognize the local or regional and melissopalynological vegetation around the beehives and nests.
Samples were collected in the Southern Brazilian region. Propolis samples became two from each of the States of Rio Grande do Sul and Santa Catarina and four from Paraná. Four geopropolis samples were from Paraná State . The 12 samples proceed from beekeepers and its apiaries (
Brazilian state | Municipalities/GPS coordinates | Bee species | Pollen grain frequencies |
---|---|---|---|
Paraná (PR) | Antonina 25˚10'05.2"S 48˚17'58.9"W | Tetragonisca angustula | AP: Cecropia (26.0%), unidentified (17.0%); IP: Alchornea (13.2%), Sapium (11.9%), Loranthaceae (3.8%) |
Tagaçaba 25˚17'59.8"S 48˚18'41.8"W | Tetragonisca angustula | AP: Cecropia (32.0%), Arecaceae (25.8%); IP: Melastomataceae (3.5%), Sapium (11.9%) | |
Melipona quadrifasciata | PP: Melastomataceae (75.5%); IP: Matayba (6.8%); Myrcia (4.0%); Mimosa scabrella (3.7%); | ||
Potinga 25˚45'24.9"S 50˚38'34.2"W | Melipona mondury | PP: Melastomataceae (50.6%); IP: Matayba (13.2%), Alchornea (9.6%), Myrcia (6.0%), Asteraceae (4.8%) | |
São José da Boa Vista 23˚54'47.7"S 49˚38'37.1"W | Apis mellifera | AP: Eucalyptus (38.4%), Asteraceae (17.9%); IP: Tapirira (8.7%), Arecaceae (9.5%), Psychotria (10.2%), Paullinia (6.4%), Solanum (5.7%) | |
São Mateus do Sul 25˚52'40.0"S 50˚22'34.0"W | Apis mellifera | AP: Asteraceae (33.5%), Eucalyptus (21.7%); IP: Tapirira (11.7%), Arecaceae (13.4%) | |
Wenceslau Brás 23˚52'18.7"S 49˚47'32.9"W | Apis mellifera | PP: Asteraceae (46.7%), Eucalyptus (32.1%); IP: Arecaceae (10.2%), Solanaceae (7.3%) | |
Ivaí 25˚00'43.5"S 50˚50'44.0"W | Apis mellifera | PP: Asteraceae (71.8%); IP: Arecaceae (12.8%), Eucalyptus (13.5%) | |
Santa Catarina (SC) | Içara 28˚44'22.7"S 49˚19'27.0"W | Apis mellifera | AP: Eucalyptus (42.1%), Asteraceae (22.5%); IP: Fabaceae (10.3%), Myrcia (10.7%) |
Anitápolis 27˚54'01.7"S 49˚08'04.5"W | Apis mellifera | AP: Eucalyptus (34.0%), Asteraceae (18.3%); IP: Arecaceae (10.1%), Cyperaceae (5.3%), Euphorbiaceae (3.4%), Melastomataceae (3.1%), Mimosa verrucosa (7.2%), Solanum (3.4%) | |
Rio Grande do Sul (RGS) | Pelotas 31˚44'56.3"S 52˚13'49.3"W | Apis mellifera | AP: Asteraceae (29.5%), Eucalyptus (27.2%); IP: Anacardiaceae (7.8%), Mimosa verrucosa (10.9%), Theaceae (3.5%) |
Taquara 29˚40'59.7"S 50˚46'28.2"W | Apis mellifera | PP: Eucalyptus (50.3%); AP: Asteraceae (15.9%), Solanum (15.6%), IP: Psychotria (4.3%) |
Extraction of pollen grains and other structured particles follows the technique described in [
Considering the three States of the Southern Brazilian region, the eight propolis samples contained mainly predominant or accessory pollen grains of Eucalyptus (13.5% - 50.3%) and of several species of Asteraceae (15.9% - 71.8%) (
Trichomes and plant tissue fragments occurred in all propolis samples and have to be detected before acetolysis was applied. Some samples presented few amorphous organic materials.
The four geopropolis samples analyzed proceeded from the State of Paraná. Two were collected by Tetragonisca angustula (“jataí”) and two by Melipona quadrifasciata (“mandaçaia”) and M. mondury (“uruçu-amarela”) bees (
Samples of the State of Paraná | |||||
---|---|---|---|---|---|
Propolis (Apis) | Geopropolis (Meliponini) | ||||
Taxon | Number of samples | Percentages (%) | Taxon | Number of samples | Percentages (%) |
Asteraceae | 4/4 | 17.9 - 71.8 | Asteraceae | 1/4 | 4.8 |
Eucalyptus | 4/4 | 13.5 - 38.4 | - | - | - |
Arecaceae | 3/4 | 9.5 - 13.4 | Arecaceae | 1/4 | 25.8 |
Tapirira | 2/4 | 8.7 - 11.7 | - | - | - |
Psychotria | 1/4 | 10.2 | - | - | - |
Solanaceae | 1/4 | 7.3 | - | - | - |
Paullinia | 1/4 | 6.4 | - | - | - |
Solanum | 1/4 | 5.7 | - | - | - |
- | - | - | Melastomataceae | 3/4 | 3.5 - 75.5 |
- | - | - | Cecropia | 2/4 | 26.0 - 32.0 |
- | - | - | Alchornea | 2/4 | 9.6 - 13.2 |
- | - | - | Matahyba | 2/4 | 6.8 - 13.2 |
- | - | - | Sapium | 2/4 | 11.9 |
- | - | - | Myrcia | 2/4 | 4.0 - 6.0 |
- | - | - | Loranthaceae | 1/4 | 3.8 |
- | - | - | Mimosa scabrella | 1/4 | 3.7 |
Predominant taxon was a Melastomataceae pollen type (Tibouchina-like) (50.6% - 75.5%) in the two samples produced by Melipona. Nevertheless, T. angustula samples presented predominant and accessory pollen grains of Cecropia (32.0 and 26.0%), and Arecaceae (25.8%). Additional taxa of lower participation in geopropolis samples of T. angustula belong to Alchornea, Sapium and Loranthaceae; the lowest frequency was of Melas- tomataceae in one sample of Tragaçaba. The Melipona geopropolis samples showed no accessory pollen beside several taxa of low participation (
All geopropolis samples presented hyphae and spores of fungi and sand, clay, mud or similar inorganic compounds, amorphous organic material (
Brazilian states | Municipalities | Bee species | Trichomes | Hyphae and spores of fungi | Sand/Clay/ mud fragments | Amorphous organic material |
---|---|---|---|---|---|---|
Paraná (PR) | Antonina | Tetragonisca angustula | - | +++ | + | + |
Tragaçaba | Tetragonisca angustula | - | + | + | ++++ | |
Tragaçaba, | Melipona quadrifasciata | - | + | + | ++++ | |
Potinga | Melipona mondury | - | + | +++ | +++ | |
São José da Boa Vista | Apis mellifera | ++++ | - | - | - | |
São Mateus do Sul | Apis mellifera | ++++ | - | - | - | |
Wenceslau Brás | Apis mellifera | ++++ | - | - | - | |
Ivaí | Apis mellifera | ++++ | - | - | - | |
Santa Catarina (SC) | Içara | Apis mellifera | ++ | - | - | ++ |
Anitapolis | Apis mellifera | +++ | - | - | - | |
Rio Grande do Sul (RGS) | Pelotas | Apis mellifera | ++++ | - | - | + |
Taquara | Apis mellifera | ++ | - | - | ++ |
A total of 22 taxa could be identified in the propolis and geopropolis samples (
The eight samples of propolis collected in the Southern Brazilian macro-region presented pollen grains belonging to 16 taxa of three or more percent frequency (
Palynological analysis of propolis samples of several Brazilian regions Brazil emphasized that predominant plant taxa in the Southern and Southeast regions comprised mainly Eucalyptus species, followed by Asteraceae [
By analogy to propolis samples from Baccharis dracunculifolia analysed by [
The four samples of geopropolis showed pollen grains belonging to 10 taxa of three or more percent frequency (
Nascimento et al. [
The Eucalyptus pollen type was frequently observed in geopropolis samples of Southeast Brazil [
Cecropia pollen type was found as secondary pollen in the honey samples from M. quadrifasciata in the Paraná State [
Pollen grains of the Asteraceae family firstly and of Eucalyptus species secondly characterized the propolis samples studies in the Southern Brazilian region. This fact is strongly different from the singles bee geopropolis samples, which showed mainly Melastomataceae and Cecropia pollen grains. Tibouchina and Miconia (Melasto- mataceae) and Cecropia (Cecropiaceae) pollen types were observed recently in honey samples of Melipona quadrifasciata and M. mondury [
Eucalyptus pollen grains may indicate human activities next to the apiaries. This genus is extensively cultivated in the Southeast and Southern Brazil . Pollen grains have been found as dominant and secondary pollen in propolis samples of the three states in Southern Brazil [
Amorphous organic material, soil or sand and fungi in the geopropolis samples studied of both, Tetragonisca and Melipona bees, were not bee species specific.
Patches of plant resins sometimes resist acetolysis procedure and were detected in the slides of Paraná state geopropolis. [
Plant tissue fragments and trichomes were introduced into propolis when the bees (Apis) cut the top of plant buds, using its mandibles, in order to open resin channels [
Depending upon locality, climatic condition and season, the vegetation investigated by Apis around the apiaries is not the same as the investigated by stingless bees, nevertheless some plant taxa may occur in common. Propolis and geopropolis are always bee species dependant.
In conclusion, the pollen grain spectra do not distinguish exactly between propolis of Apis and geopropolis of stingless bees, but the accessory elements are conclusive, mainly trichomes and mineral elements.
Financial support: Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) to the first author (Fellowship process number 304067/2013-0).
Ortrud Monika Barth,Alex da Silva de Freitas, (2015) Palynology as a Tool to Distinguish between Propolis and Geopropolis: Southern Brazilian Samples. Open Access Library Journal,02,1-10. doi: 10.4236/oalib.1102217