Pollen in honey of Melipona scutellaris L

Palynology

ISSN: 0191-6122 (Print) 1558-9188 (Online) Journal homepage: http://www.tandfonline.com/loi/tpal20

Pollen in honey of Melipona scutellaris L. (Hymenoptera: Apidae) in an Atlantic rainforest area in Bahia, Brazil Vanessa Ribeiro Matos & Francisco de Assis Ribeiro dos Santos To cite this article: Vanessa Ribeiro Matos & Francisco de Assis Ribeiro dos Santos (2017) Pollen in honey of Melipona scutellaris L. (Hymenoptera: Apidae) in an Atlantic rainforest area in Bahia, Brazil, Palynology, 41:1, 144-156, DOI: 10.1080/01916122.2015.1115434 To link to this article: https://doi.org/10.1080/01916122.2015.1115434

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Palynology, 2017 Vol. 41, No. 1, 144156, http://dx.doi.org/10.1080/01916122.2015.1115434

Pollen in honey of Melipona scutellaris L. (Hymenoptera: Apidae) in an Atlantic rainforest area in Bahia, Brazil Vanessa Ribeiro Matos* and Francisco de Assis Ribeiro dos Santos Departamento de Bot^ anica, Laborat orio de Micromorfologia Vegetal, Universidade Estadual de Feira de Santana, 44036-900 Feira de Santana, Bahia, Brasil. Uru¸c u (Melipona scutellaris L.) is an example of a native stingless bee typical of northeastern Brazil. It stands out due to its high-quality honey and for being responsible for 40 to 90% of the pollination of native plant species. A total of 37 honey samples were analysed, collected between July 2012 and June 2014 in a meliponary located in the municipality of Entre Rios, in the state of Bahia, Brazil. Honey samples were diluted in distilled water, then in alcohol, and then were acetolysed. Climate information (precipitation and temperature) for the area was obtained during the same period of time. We found 102 pollen types, of which 71 were identified as belonging to 36 plant families. Fabaceae was the most representative family with 15 types, followed by Myrtaceae with seven types. Myrtaceae was highly significant due to the high distribution frequency and dominance of the Eucalyptus type. The Miconia type (Melastomataceae), highly polliniferous, also stood out in our analysis with a high distribution frequency and dominance in some of the sampled months. The precipitation index of the second year of study (August 2013 to June 2014) was much greater than the first year (July 2012 to June 2013), being proportional to the increase of pollen diversity in the same period. Keywords: melissopalynology; bee products; native plants; precipitation index; Bahia, Brazil

1. Introduction Social bees are important pollinating agents, besides being the most significant species for honey production, among which Melipona scutellaris L. is especially noteworthy (Carvalho et al. 1999; Mendes et al. 2009). Added to that, these bees produce an important food from flower nectar, i.e. honey. It is stored in pots and its nutritional composition varies according to the local flora, which in turn is influenced by the edaphoclimatic conditions (Anacleto et al. 2009; Mendes et al. 2009). Native bees (Meliponines) are responsible for around 40 to 90% of the pollination of native plant species (Kerr et al. 1996). In northeastern Brazil, the species known as uru¸c u (Melipona scutellaris L.) is characterised by the production of high-quality honey, good hygiene and easy domestication (Rodrigues et al. 2008). Recently there has been an increase in the sale of native bee products; this, coupled with high demand, has turned meliponine honey into a commodity much more expensive than Apis mellifera L. honey. However, the African bee possesses a much larger population, so its production is numerically superior, making it commercially much more profitable (Souza et al. 2009). One of the ways of identifying the bee-visited flora is by analysing the pollen types present in the produced honey. This is only possible since, while *Corresponding author. Email: [email protected] Ó 2016 AASP The Palynological Society

Published online 04 Apr 2016

visiting the flowers, the bees contaminate themselves with pollen grains. Part of this pollen is mixed into the honey when the nectar is regurgitated into the pot in the hive (Santana et al. 2011). Thus, melissopalynological analysis is a useful tool for identifying the regional bee flora, and may indicate its potential for apiculture (Crompton & Wojtas 1993; Silva & Santos 2014). In Bahia, despite its diverse flora and its elevated potential for apiculture, there is little information regarding the bee flora (Moreti et al. 2000). Recently, there have been some efforts to infer the botanical and geographical origin of meliponine honey in Bahia. Nevertheless, most of these studies have been done in areas inside the Caatinga domain, while few studies were conducted in areas inside the Atlantic rainforest domain (Alves et al. 2006; Demartelaere et al. 2010; Novais et al. 2013). Hence, the present work aimed to identify and to characterise (occurrence, habit type and floral resources) the pollen grains contained in the Melipona scutellaris L. honey samples. The samples were obtained from a meliponary located in an Atlantic rainforest region in the state of Bahia, Brazil. The purpose is to provide information that contributes to a better understanding of the regional bee flora.

Palynology

145

was not possible to collect honey samples during August 2012 and July 2013 due to intense rain.

Figure 1. Location of the study area: (A) state of Bahia, Brazil; (B) micro-region of Entre Rios (highlighted in grey).

2. Materials and methods 2.1 Study area The study was developed in the municipality of Entre Rios, Bahia, Brazil (Figure 1). The chosen meliponary was Fazenda S~ ao Lucas (12 010 78600 S, 38 040 36600 W), located in an area inside the Atlantic rainforest domain. The area contains well-preserved secondary vegetation, which is maintained to serve as a foraging area for the bees and consequently increase the local production of bee products, especially honey. The municipality of Entre Rios is located at roadway BR110, between Salvador and Alagoinhas. In 2013, the region produced 7570 kg of honey from bees (Apis mellifera and native bees), generating an income of 34000 reais in the municipality (IBGE 2002).

2.2

Sampling

Two hives of M. scutellaris (A and B), placed next to one to another, were selected for analyses, from which monthly samples were collected from the honey pots, whenever they were available. Care was taken to remove all contents of these honey pots, thus making sure that there were new honey pots next month. A total of 37 samples, during a period of 24 months (June 2012 to June 2014), were obtained from the honey pots of both boxes. During the first year of study (June 2012 to June 2013), we collected a total of 21 samples. During the second year (August 2013 to June 2014) there was a decline in the obtained honey samples, with a total of just 16. It

2.3 Pollen analysis The honey samples (10 g) were diluted in distilled water (Louveaux et al. 1978), then in alcohol (Jones & Bryant Jr. 2004), and finally they were acetolysed (Erdtman 1960) and mounted on glass slides with glycerin jelly. For the determination of classes and frequencies of pollen types in the honey samples, 500 pollen grains were counted per sample. The adopted frequency classes were established by Louveaux et al. (1970, 1978): predominant pollen (> 45%), secondary pollen (1645%), important minor pollen (315%), minor pollen (13%) and trace pollen (< 1%). The distribution frequency of the pollen types (taxa) followed Jones & Bryant Jr. (1996): very frequent (> 50%), frequent (2150%), infrequent (1020%) and rare (< 10%). For the qualitative analyses, the pollen types found in the honey samples had their botanical affinity recognised through comparison with pollen slides deposited at the Palynotheca of Laborat orio de Micromorfologia Vegetal, Universidade Estadual de Feira de Santana (LAMIV/UEFS), with help of specialised literature, and conforming with the recommendations of Santos (2011). All pollen types were photographed. 2.4 Macroclimatic data collection The macroclimatic data (i.e. temperature and precipitation), referring to the whole study period, were obtained from the conventional meteorological station of Alagoinhas, through the Instituto Nacional de Meteorologia (INMET). The Alagoinhas conventional meteorological station was chosen, since it was the nearest station to the study area. 2.5

Habit and floral resources

The habit classification referring to the taxa inferred through the pollen types was based on Marchant et al. (2002), Brito et al. (2006), Queiroz (2009), Modro et al. (2011) and Alves & Santos (2014). We followed the studies of D orea (2007), Luz et al. (2007) and Concei¸c ~ao (2013) for identifying the floral resources that each taxon provides the bees. 2.6

Statistical analysis

The analysis of the similarity between samples was done using the software PRIMER-E (Plymouth Routines In Multivariate Ecological Research), version 6.1.6 (Clarke & Gorley 2006). The chosen similarity coefficient was the Jaccard (j), since it does not

146

V. R. Matos and F. de A. Ribeiro dos Santos

consider shared absence as evidence for similarity. Thus, only the data regarding the presence or absence of pollen types was considered. The software STATISTICA (Data Analysis Software System) version 10 (StatSoft 2011) was used to correlate precipitation with pollen type diversity during the studied period, using a simple regression. 3. Results In the 37 studied honey samples, a total of 102 pollen types were found, out of which 71 were identified as belonging to 36 plant families (Tables 1 and 2). The most representative family in our study was Fabaceae, with 15 pollen types, followed by Myrtaceae, with seven types. In our first study year, in the 21 obtained honey samples, we identified 49 types, belonging to 23 plant families (Table 1). During the second study year, even with a smaller number of samples (16), we identified a greater number of pollen types: 61, belonging to 31 plant families (Table 2). The Eucalyptus type (Myrtaceae; Plate 1, figure 10) was classified as the predominant type in 14 samples (Tables 1 and 2). This pollen type had a distribution frequency of greater than 95% during the first study, while during the second year it was present in all analysed samples (Figure 2). The species belonging to this type are a source of both nectar and pollen to the bees. Beside the Eucalyptus type, other pollen types belonging to species of this family were important nectar sources to the uru¸c u bee during the study period, since related pollen types were identified especially as secondary pollen. We identified three Myrcia pollen types (Plate 1, figures 1113), and the Eugenia type (Tables 1 and 2). The types referring to the taxa of Myrcia also presented a high distribution frequency during the whole study period (Figure 2). The species belonging to Sapindaceae are considered excellent nectar sources for the bees. The Serjania pollen type (Plate 1, figure 18) was predominant in the September and November 2012 samples (Table 1), and secondary in the October 2013 sample (Table 2). Added to that, it had a high distribution frequency in the last 12 months of the study period (Figure 3). Other pollen types belonging to Sapindaceae were classified as secondary pollen: the Cupania type (Plate 1, figure 17: September 2013) and the Paullinia racemosa type (May 2014; Table 2). The Miconia type (Melastomataceae; Plate 1, figure 9) represents a group of plants that is mainly a pollen source for the bees. However, it was identified in the uru¸c u honey samples with a distribution frequency of higher than 80% in the first study year and of 100% in the second year (Figure 2). Pollen types corresponding to Fabaceae were also important sources of floral

resources for the bees in the studied area. Among them we can highlight the Mimosa pudica type (Plate 1, figure 8), Mimosa arenosa type (Plate 1, figure 7) and Piptadenia type. The Simarouba amara type (Simaroubaceae; Plate 1, figure 19) was found as predominant pollen in the October 2012 and 2013 honey samples. The species referred to this type is considered to be nectariferous and polliniferous. Regarding the floral resources available to Melipona scutellaris in the studied Atlantic rainforest area, the nectariferous predominated in the pollen spectra (30 taxa) when compared to the polliniferous (18 taxa). Nevertheless, during the 24 studied months we recorded the occurrence of plants (22 taxa) that provided both resources (nectariferouspolliniferous), whereas seven taxa were identified as resiniferous and just three taxa were identified as providing essential oils (Tables 1 and 2). According to the habit, the species belonging to the identified pollen types fall into the following categories: tree (A), shrub (S), vine (C), liana (L) and herb (H). During the first study year, the most frequent habit type was tree (35 taxa), followed by shrub (13 taxa), herb (12 taxa), liana (two taxa) and vine (one taxon) (Table 1). During the second year, the arboreous habit continued to be the most represented (39 taxa), followed by shrub (18 taxa), herbaceous (17 taxa) and vine (four taxa), and the least represented being liana with two taxa (Table 2). The average temperature in the region during the study period varied from 22 to 28 C. The highest recorded temperature was 34 C in January and February 2013, while the lowest was 18 C in August 2012. The precipitation index in the second study year (87.2 mm3) was higher than that in the first year (72.4 mm3), which corroborates the increase in the pollen diversity of the examined samples, during the same period. Nevertheless, in April 2013 we recorded the second highest precipitation volume in the region (190 mm3), while in the second half of 2012 rainfall was very low in the region; hence, we recorded the lowest precipitation volume in the region (0.3 mm3) (Figure 3). There was no significant correlation between precipitation and the number of identified pollen types. According to the statistical test applied to both hive A (F D 1.61, p < 0.21, r2 D 0.02, b D 1.7) and hive B (F D 0.47, p < 0.5, r2 D 0.02, b D 1.4), there was no relationship between these two variables (pollen type and precipitation). Thus, other factors may have contributed to the quantitative variation of pollen types in the study area. The highest similarity index (ca. 67%) occurred between the samples from November 2012 (A & B; Figure 4-1G). These samples shared 16 pollen types. The samples from December 2012 (A & B;

R

R VF F R

R R F

R LF

R

F

F

LF

F

F F R R F VF VF F R F LF

LF

Anacardiaceae Astronium Schinus terebinthifolius Spondias tuberosa Tapirira guianensis

Arecaceae Arecaceae Bactris ferruginea Syagrus coronata

Asteraceae Mikania Vernonanthura

Bromeliaceae Bromeliaceae

Buseraceae Protium heptaphyllum

Cannabaceae Celtis

Commelinaceae Commelina

Cyperaceae Rhynchospora cephalotes

Fabaceae Chaemecrista ramosa Fabaceae I Fabaceae II Fabaceae IIII Inga Mimosa arenosa Mimosa pudica Mimosa tenuiflora Plathymenia Piptadenia Senna

Malpighiaceae Byrsonima sericea

FD

Amaranthaceae Alternathera ramosissima

Pollen types

Samples

B

A

A

B

A

B

A

B

A

B

A

B

A

B

A

B

A

B

A

B

I C

C S I C

C

C

I

C

C

C

C

I C

C

C

C

C S

m C

C

m

C

m

I

C

C

C

C

m

m

I C

I

I C m

C

C

m C C

m

m

m m

m m

C

I

C

S C C

S

m I m C

C

S C

C

C

m C

C

C

I

I

I

I C

C C

m

I

C

m

C

C

m I

C

C

m

C

C

C

C

m

C

C

m

C

C C C

C

C C

m

C

I m

C

C

C

C

C

m

C m m

C

C

C C

C m

C

C

C

C

C

m C

C

C

C

C

C

C m I S

C

C

C

C

C

(07/12) (07/12) (09/12) (10/12) (10/12) (11/12) (11/12) (12/12) (12/12) (01/13) (01/13) (02/13) (02/13) (03/13) (03/13) (04/13) (04/13) (05/13) (05/13) (06/13) (06/13)

A

O

P N/P N/P N/P N/P N/P P P N N/P N/P

P

N

P

N/R

N

P N/P

N/P P P

N/P/R N/P/R N/R N/R

N

FR

(continued)

A

A/S A/S/H A/S/H A/S/H A A H A A A S

H

H

A/S

A

A/H/L

C S/H

A A A

A A A A

H

H

Table 1. Frequency of pollen types in the honey samples collected from pots of M. scutellaris L. in an area of Atlantic rainforest in the state of Bahia (Brazil), during the first study year (July 2012 to June 2013). Frequency classes: D predominant pollen (> 45%); S secondary pollen (1645%); I important minor pollen (315%); m minor pollen (13%); C trace pollen (< 1%). Numbers in brackets D (month/year).

Palynology 147

LF

VF F VF VF VF R F

LF

F

LF

R R R

R

R R F

F

F

F

Moraceae Ficus hirsuta

Myrtaceae Eucalyptus Eugenia Myrcia I Myrcia II Myrcia III Myrtaceae Psidium

Plantaginaceae Angelonia

Poaceae Poaceae

Polygalaceae Polygala paniculata

Rubiaceae Borreria verticillata Guettarda viburnoides Mitracarpus

Rutaceae Citrus

Sapindaceae Cupania Paullinia racemosa Serjania

Simaroubaceae Simarouba amara

Solanaceae Solanum paniculatum

Urticaceae Cecropia

B

A

A

B

A

B

A

B

A

B

A

B

A

B

A

B

A

B

A

B

(2.8/3) 21

C

I C

C

10

C

m C

C I C

C

D

m

S

S

9

D

C C

C

I

m

D

C

C

m

m

I

m

I

C

D

C

I

I

m

C

C

m

C

C

I I

C I

S

I m

C m

I

m

I

C

C

C

m I C S I

m

I

m

I

C

C

I D I I S

I

C

C

m

m

C

C

m C S I I

m

m

S C I I I C

m

(9.0/6) (6.4/1) (1.4/1) (0.4/2) (0.6/1) (0.2/1) (2.2/3) (2.2/3) 22 13 22 21 20 20 24 24

C

m

S

m

C

C

S

10

C

C C

D

C

5

C

D

m

8

D C S S I

11

C

C

C m C

D

8

C

C I I

D

C

C

C

C

m I I

D

C

m

m I C

D

C

C

(1.2/1) (0.2/1) (0.4/1) 18 17 15

C

C

C

I I m

D

m

C

13

C

C

C

C I

D

17

C

I C m I C

D

(07/12) (07/12) (09/12) (10/12) (10/12) (11/12) (11/12) (12/12) (12/12) (01/13) (01/13) (02/13) (02/13) (03/13) (03/13) (04/13) (04/13) (05/13) (05/13) (06/13) (06/13)

A

A

A

A

A/S A L

A

H A S

H

H

S/H

A A/S A A A A/S A

A

A/S

H

N/P/R

P

N/P

N N N

N

N/P N/P N

N

P

N/O

N/P N/P N/P N/P N/P N/P P

N/O

P

FR

FD (frequency of distribution): R D rare; LF D low frequency; F D frequent; VF D very frequent. H (habit): A D arboreal; S D shrubby; H D herbaceous; L D liana; C D climber. FR (floral resource): N D nectar; P D pollen; R D resin; O D oil. A/B (boxes of M. scutellaris). Months: 01 D January; 02 D February; 03 D March; 04 D April; 05 D May; 06 D June; 07 D July; 08 D August; 09 D September; 10 D October; 11 D November; 12 D December.

Pollen types not identified (%/no.) Total of pollen types (no.)

VF

FD

Melastomataceae Miconia

Pollen types

Samples

Table 1. (Continued )

148 V. R. Matos and F. de A. Ribeiro dos Santos

R R VF F R

LF

LF LF

VF

LF

LF

R

F

R F

LF F F LF R VF LF VF R VF R VF F

Arecaceae Syagrus coronata

Asteraceae Mikania Vernonanthura

Buseraceae Protium heptaphyllum

Cannabaceae Celtis

Capparaceae Colicodendron

Convolvulaceae Jacquemontia

Cyperaceae Rhynchospora cephalotes

Euphorbiaceae Croton camprestris Euphorbia

Fabaceae Acacia Chaemecrista ramosa Inga Fabaceae III Machaerium Mimosa arenosa Mimosa misera Mimosa pudica Mimosa quadrivalvis Mimosa tenuiflora Plathymenia Piptadenia Senna

FD

Anacardiaceae Astronium Schinopsis Schinus terebinthifolius Spondias tuberosa Tapirira guianensis

Pollen types

Samples

m I

I m I

I

m

C

C

I C

m

(08/13)

B

I

C

C

M C

(09/13)

A

m

C

C

I

I

C

C

C

C

C

C C

C

C

(10/13)

B

C

C

C

C

C

C

(10/13)

A

m

I

C

C

C

I C

(11/13)

A

I m I

1 C C I C

I

C

I

C

C C

(11/13)

B

m

C

I

C

I

(12/13)

A

C m

m

I C I

C

C

C

C

m

(12/13)

B

S

m

I

C

m

C

(01/14)

B

m

I

I

I

C I

C

(02/14)

B

m C

m

m m m

m

m

C

C

(03/14)

B

C

m

m

I

C

(04/14)

A

I

C

m

C

C

C

m

(04/14)

B

m C

m

C

C

C

(05/14)

A

m C

D

C

C

(05/14)

B

m

m

m

C

C

m C

C

m

C

C

(06/14)

B

P P N/P N/P N N/P P P P P N N/P N/P

N/P N

P

N

N/P

P

N/R

P N/P

P

N/R N/R N/P/R N/R N/R

FR

(continued)

A A/S A A/S/H A A H H H A A A S

S S/H

H

L

A

A/S

A

C S/H

A

A A A A A

H

Table 2. Frequency of pollen types in the honey samples collected from pots of M. scutellaris L. in an area of Atlantic rainforest in the state of Bahia (Brazil), during the second study year (August 2013 to June 2014). Frequency class: D predominant pollen (> 45%); S secondary pollen (1645%); I important minor pollen (315%); m minor pollen (13%); C trace pollen (< 1%). Numbers in brackets D (month/year).

Palynology 149

F LF

LF

F

LF VF

LF

LF

VF VF VF VF VF R F

R

R

R

F

F

F

F

Loranthaceae Loranthaceae

Malpighiaceae Byrsonima sericea

Melastomataceae Clidemia Miconia

Meliaceae Trichilia

Moraceae Ficus hirsuta

Myrtaceae Eucalyptus Eugenia Myrcia I Myrcia II Myrcia III Myrtaceae Psidium

Nyctaginaceae Nyctaginaceae

Passifloraceae Passiflora

Phytolacaceae Microtea

Phyllanthaceae Phyllanthus

Plantaginaceae Angelonia

Poaceae Poaceae

Rhamnaceae Ziziphus

FD

Lamiaceae Hyptis Salvia

Pollen types

Samples


I

C

C

C

C

m

C

C

M M

I

I

(10/13)

A

I C m m

I

C

S

C

(09/13)

A

S C I S

C

C I

(08/13)

B

C

C

S C m m C

I

(10/13)

B

C

C

I m m S I C C

C

C

I S

C

m C

(11/13)

A

C

C

m I m I I

m

C I

C

(11/13)

B

m

C C C m I

D

C

C

(12/13)

A

I

C

m

C

C

I I m I I

I

I

C

C

m C

(12/13)

B

I

I

C

m m m S S

m

C

(01/14)

B

m

C

m I

D

C

(02/14)

B

m

C

D C C I

m

(03/14)

B

I C

C S m

C

D

m

C

(04/14)

B

D

8

(04/14)

A

m S

S

I

(05/14)

A

C

I

S

S

(05/14)

B

C I C

I

D

m

(06/14)

B

N

P

N/O

N/R

N

N

N

N/P N/P N/P N/P N/P N/P P

N/O

N

P P

O

N

N N

FR

(continued)

A

H

S/H

S/H

H

C

A/S/H/C

A A/S A A A A/S A

A

A

S A/S

A

H/C

S/H A/S/H

H

150 V. R. Matos and F. de A. Ribeiro dos Santos

R

F LF VF

LF

VF

VF

F

Salicaceae Casearia

Sapindaceae Cupania Paullinia racemosa Serjania

Sapotaceae Pouteria

Simaroubaceae Simarouba amara

Solanaceae Solanum paniculatum

Urticaceae Cecropia (2.6/4) 35

(0.2/1) 26

I

I

(2.0/3) 29

(0.2/1) 18

I

C

C

C

C C

(11/13)

A

C (0.2/1) 18

C

D

S

C

C

(10/13)

B

m I

(10/13)

A

S

M

(09/13)

A

m

C

C

(08/13)

B

(2.0/4) 33

I

m

C

C

(11/13)

B

14

(12/13)

A

(0.6/2) 37

C

I

m

C

m

C

I C

(12/13)

B

(0.4/2) 21

m

C

m

(01/14)

B

14

C

(02/14)

B

(0.4/1) 19

C

(03/14)

B

¡ 13

C

m

(04/14)

A

¡ 14

C

C

(04/14)

B

(0.2/1) 13

I

S

(05/14)

A

¡ 10

C

(05/14)

B

¡ 19

1

C

I

(06/14)

B

A

A

A

A

A/S A L

A/S

H S H

H

N/P/R

P

N/P

N/R

N N N

P

N/P N N

FR

scutellaris). Months: 01 D January; 02 D February; 03 D March; 04 D April; 05 D May; 06 D June; 07 D July; 08 D August; 09 D September; 10 D October; 11 D November; 12 D December.

FD (frequency of distribution): R D rare; LF D low frequency; F D frequent; VF D very frequent. H (habit): A D arboreal; S D shrubby; H D herbaceous; L D liana; C D climber. FR (floral resource): N D nectar; P D pollen; R D resin; O D oil. A/B (boxes of M.

Pollen types not identified (%/no.) Total of pollen types (no.)

F LF R

FD

Rubiaceae Borreria verticillata Mitracarpus Richardia grandiflora

Pollen types

Samples


Palynology 151

152


Plate 1. Pollen types found in the honey samples collected from pots of M. scutellaris L. in an area of Atlantic rainforest in the state of Bahia (Brazil). Figure 1. Anacardiaceae, Schinus terebinthifolius. Figure 2. Arecaceae, Syagrus coronata. Figures 34. Asteraceae: Figure 3. Mikania. Figure 4. Vernonanthura. Figure 5. Buseraceae, Protium heptaphyllum. Figure 6. Convolvulaceae, Jacquemontia. Figures 78. Fabaceae: Figure 7. Mimosa arenosa. Figure 8. Mimosa pudica. Figure 9. Melastomataceae, Miconia. Figures 1013. Myrtaceae: Figure 10. Eucalyptus. Figure 11. Myrcia I. Figure 12. Myrcia II. Figure 13. Myrcia III. Figures 1416. Rubiaceae: Figure 14. Borreria verticillata. Figure 15. Guettarda viburnoides. Figure 16. Richardia grandiflora. Figures 1718. Sapindaceae: Figure 17. Cupania. Figure 18. Serjania. Figure 19. Simaroubaceae, Simarouba amara. Figure 20. Solanaceae, Solanum paniculatum. Scale bars D 10 mm.

Palynology

153

Figure 2. Frequency of pollen types present in more than 50% of the analysed honey samples collected from pots of M. scutellaris L. in an area of Atlantic rainforest in the state of Bahia (Brazil), during the period of July 2012 to June 2014.

Figure 4-2G) also presented a high similarity index (ca. 66%), with 15 pollen types shared, and samples A in May 2013 and A in April 2014 (Figure 4-3G) also presented a high similarity index (ca. 61.2%), with nine shared pollen types.

4. Discussion The pollen analysis of the honey produced by the bee M. scutellaris in an area of Atlantic rainforest, in the

state of Bahia, shows a foraging area that provides a wide variety of resources. According to Alves et al. (2006), bees from this species are considered policletic since they feed on the floral resources of different plant families, which sets them apart from the remaining stingless bees. Other studies carried out in a similar area indicated 28 to 43 pollen types (Carvalho et al. 2001; Andrade et al. 2009; Santana et al. 2011). Myrtaceae is fairly well represented in the pollen analysis of bee products. Aside from the Eucalyptus

Figure 3. Ratio between precipitation and average temperature of the area of Atlantic rainforest in the state of Bahia (Brazil) where the honey samples were collected from pots of M. scutellaris L., during the period of July 2012 to June 2014.

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Figure 4 Similarity dendrogram (Jaccard’s coefficient) between the honey samples collected from pots of M. scutellaris L. in an area of Atlantic rainforest in the state of Bahia (Brazil), highlighting key similar groups (G).

type, which indicates an important source of pollen and nectar for the bees, other types belonging to Myrtaceae are frequent in honey samples of native bees. The Psidium type was classified as a predominant pollen type in honey samples of M. scutellaris from the municipality of Catu (Bahia; Carvalho et al. 2001). On the other hand, Santana et al. (2011) also identified the Eugenia type as being present in the uru¸c u honey, while Novais et al. (2013), analysing the honey of Tetragonisca angustula from the municipality of Itaberaba, found the Myrcia pollen type in 13 analysed samples, classifying it as secondary pollen in one of them. According to Oliveira (2011), Sapindaceae is an important source of nectar for a wide range of bee species in the state of Bahia. Silva (2009), analysing flowers of a species belonging to Serjania, found M. scutellaris to be the second most frequent floral visitor. Andrade et al. (2009) found Serjania pollen in their analyses, while Santana et al. (2011) found the Cupania type, besides this type. Both types also provided resources for the uru¸c u at the Fazenda S~ ao Lucas, in the municipality of Entre Rios, during the two years this study was held. Even though Melastomataceae are not good nectar sources, pollen types belonging to this family are commonly found in honey samples produced by native bees. This may be due to contamination by pollen, stocked in feeding pots within the hive, since the group is commonly polliniferous (Alves et al. 2006).

In the studied Atlantic rainforest area, Fabaceae had great relevance in the diet of the uru¸c u, during the two studied years. Nevertheless, the Mimosa arenosa, Mimosa pudica and Piptadenia types were the most representative. In the studies carried out by Santana et al. (2011) in the semiarid region of the state of Bahia, the family stood out as a source of floral resources for M. scutellaris in the region. The M. arenosa and Piptadenia types stood out, being classified as predominant in some analysed samples, while Andrade et al. (2009) found as predominant pollen for this bee, in an area of family farming in the state of Bahia, two pollen types belonging to the genus Mimosa (M. pudica and M. quadrivalvis). Simarouba amara Aubl. (Simaroubaceae) is characteristic to secondary rainforest (Carvalho et al. 2007). It occurs in Brazil in the states within the Amazon Rainforest domain and in the states of Bahia, Cear a and Pernambuco. Its blooming begins together with the rainy season, and peaks in December (Azevedo et al. 2010). Simarouba amara is a species classified as nectariferouspolliniferous; however, its primary resource for the bees is the nectar. The arboreous and shrubby species’ greatest contribution probably occurred since the farm where the meliponary is located still possesses remnant Atlantic rainforest vegetation in good condition, even with intense expansion of Eucalyptus plantations in the

Palynology region. According to Pereira et al. (2004), the tree layer is important in the bees’ diet, since it provides nectar during the dry season. It also provides nectar during the transitional period between the dry and rainy seasons, and pollen during the rainy season. During the second study year, there was a signfiicant increase in the precipitation in the area. This may have boosted the blooming of local plants, consequently increasing the pollen diversity in the honey samples during the same period. Novais et al. (2009) and Alves (2013) indicated that climatic variables, especially precipitation, affect pollen diversity in bee products in Northeastern Brazil. 5. Conclusions In the region of Entre Rios (Bahia) in an area of Atlantic rainforest, the honeybee Melipona scutellaris, popularly known as uru¸c u, exhibited a high diversity of plant species in its diet. Making use of pollen analysis in the produced honey, during a period of 24 months, a total of 102 pollen types were found, belonging to 36 plant families. These pollen types are taxonomically related to nectariferous (42.2%), polliniferous (25.3%) and nectariferouspolliniferous plants (42.2%), and plants that produce essential oils and resins (14.08%). Fabaceae is important in the diet of Melipona scutellaris, which can be corroborated by the great number of pollen types found, especially Mimosa pudica, Mimosa arenosa and Piptadenia. Sapindaceae, highly nectariferous, stood out with the Serjania type, with an affinity for lianescent plants. This pollen type was the predominant one in some months and also had a high distribution frequency during some times of the year. In the study area, there is a large Eucalyptus plantation for wood extraction, which may explain the occurrence of this pollen type in the honey samples. The species belonging to this genus are also considered invasive and highly nectariferous. However, the Miconia type (Melastomataceae) stood out in our analysis with a high frequency and dominance in some of the sampled months, despite its affinity with taxa with low nectar production. In the same way, the Simarouba amara type produces nectar and also provides the bees with a great amount of nectar. The high representation of these types in the pollen spectra, correlating to taxa considered to be polliniferous, could be produced by a contamination, caused by the bees themselves. Even with the high occurrence of types during the rainy season, there was no significant correlation between precipitation and the pollen spectra. Other factors like the habit of the visited plants, and the behavior of the studied bee, may have contributed to this high diversity of pollen types during the rainy season.

155

Acknowledgements Many thanks are due to the Plant Micromorphology Laboratory (State University of Feira de Santana) for facilities, to Coordena¸c ~ ao de Aperfei¸c oamento de Pessoal de Nıvel Superior (CAPES) for giving a grant to the first author, and to the Conselho Nacional de Desenvolvimento Cientıfico de Tecnol ogico (CNPq) for supporting FARS (# 303862/2013-0).

Disclosure statement No potential conflict of interest was reported by the authors.

Author biographies VANESSA R. MATOS has a degree in biological sciences from the UNIJORGE, and a master’s degree in botany from the State University of Feira de Santana. Currently, she is a PhD student comparing the pollen flow in M. scutellaris L. and Apis mellifera L. products in a rainforest region in the state of Bahia, Brazil. She has experience in palynology of pollen and pollen morphology. FRANCISCO DE ASSIS R. DOS SANTOS has a degree in biological sciences from the Federal University of Bahia (1987), a master’s degree in botany from the Federal Rural University of Pernambuco (1993) and a PhD in sciences (botany) from the University of S~ ao Paulo (1998). He developed a postdoctoral program of study in palynology at the Escuela Nacional Biological Sciences (National Polytechnic Institute, Mexico, DF). He is currently a full professor at the State University of Feira de Santana. His research is on all aspects of palynology studies.

ORCID Francisco de Assis Ribeiro dos Santos 0002-9246-3146

http://orcid.org/0000-

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