Pollen analysis of honeys from Apis mellifera and Tetragonisca fiebrigi (Hymenoptera: Apidae) in the Upper Paraná Atlantic Forest, Argentina

Fabio Fernando Flores Norma Inés Hilgert Fernando Zamudio Fernanda Fabbio Liliana Concepción Lupo About the authors

Abstract

Nectariferous and polliniferous resources are key to the survival of social bees, so identifying the origin sources allows the implementation of management actions aimed at a greater supply of nutrients for the colonies. Besides, defining the floral origin of honeys contributes to their characterization and commercialization. The objective of our work was to identify the plants that provide nectar to the bees Apis mellifera and Tetragonisca fiebrigi in northern Misiones, through palynological analysis of honey samples collected between 2006 and 2008. Both bees showed a polylectic foraging habit. Richness of pollen types per sample ranged between 10 and 34 (mean = 20.5 ± 7.7) for A. mellifera, and between 13 and 43 (mean = 24.8 ± 7.1) for T. fiebrigi. The 15 most abundant pollen types in the honeys of A. mellifera were, in decreasing order of importance, Euphorbiaceae, Euterpe edulis, Holocalyx balansae, Calyptranthes concinna-type, Actinostemon, Salix, Ruprechtia laxiflora, Myrcianthes pungens-type, Thinouia mucronata, Allophylus edulis, Ilex, Syagrus romanzoffiana, Gouania latifolia-type, Parapiptadenia rigida, and Baccharis-type, whereas in the honeys of T. fiebrigi the most important pollen types included S. romanzoffiana, Schinus weinmannifolius-type, Baccharis-type, H. balansae, E. edulis, Rhamnaceae, Citrus, Leonurus japonicus, G. latifolia-type, A. edulis, Gomphrena perennis-type, Pouteria gardneriana, P. rigida, Zanthoxylum, and Actinostemon.

Key words
honeybee; Melissopalynology; melliferous flora; stingless bee

Resumen

Los recursos nectaríferos y poliníferos son clave para la supervivencia de las abejas sociales, por lo que identificar las fuentes de origen permite implementar acciones de manejo tendientes a un mayor suministro de nutrientes para las colonias. Asimismo, definir el origen floral de las mieles contribuye a la caracterización y comercialización de las mismas. El objetivo de nuestro trabajo fue identificar los recursos vegetales que proporcionan néctar a las colmenas de Apis mellifera y Tetragonisca fiebrigi en el norte de Misiones, a partir del análisis polínico de las muestras de miel colectadas entre 2006 y 2008. Ambas especies mostraron un hábito de alimentación poliléctico. La riqueza de tipos polínicos por muestra osciló entre 10 y 34 (media = 20,5 ± 7,7) para A. mellifera, y entre 13 y 43 (media = 24,8 ± 7,1) para T. fiebrigi. Los 15 tipos polínicos más abundantes en las mieles de A. mellifera fueron, en orden decreciente de importancia, Euphorbiaceae, Euterpe edulis, Holocalyx balansae, tipo Calyptranthes concinna, Actinostemon, Salix, Ruprechtia laxiflora, tipo Myrcianthes pungens, Thinouia mucronata, Allophylus edulis, Ilex, Syagrus romanzoffiana, tipo Gouania latifolia, Parapiptadenia rigida y tipo Baccharis, mientras que en las mieles de T. fiebrigi incluyeron a S. romanzoffiana, tipo Schinus weinmannifolius, tipo Baccharis, H. balansae, E. edulis, Rhamnaceae, Citrus, Leonurus japonicus, tipo G. latifolia, A. edulis, tipo Gomphrena perennis, Pouteria gardneriana, P. rigida, Zanthoxylum y Actinostemon.

Palabras clave
abeja melífera; Melisopalinología; flora melífera; abejas sin aguijón

Introduction

Nectariferous and polliniferous resources are key to the survival of social bees and their colonies (Michener 2007Michener CD (2007) The bees of the world. 2nd ed. The Johns Hopkins University Press, Baltimore. 972p.). The use of these resources can be analysed by observing the flora and its visitors (Souza et al. 2016Souza CS, Aoki C, Ribas A, Pott A & Sigrist MR (2016) Floral traits as potential indicators of pollination vs. theft. Rodriguésia 67: 309-320.) or by pollinic analysis of food sources (honey and pollen) stored in nests (Vossler 2019Vossler FG (2019) Pollen diet assessment and flower association in Melipona orbignyi and recommendations on management and conservation of stingless bees in the Chaco dry forest of South America. Apidologie 50: 391-413.). The pollen present in the nests can be a botanical and geographical indicator of their provenance (Barth 1989Barth OM (1989) O pólen no mel brasileiro. Editorial Luxor, Rio de Janeiro. 150p.; Soares et al. 2017Soares S, Amaral JS, Oliveira MBPP & Mafra I (2017) A comprehensive review on the main honey authentication issues: production and origin. Comprehensive Reviews in Food Science and Food Safety 16: 1072-1100.; Alves & Santos 2018Alves RF & Santos FAR (2018) Pollen foraged by bees (Apis mellifera L.) on the Atlantic Forest of Bahia, Brazil. Palynology 43: 523-529.; Bandeira & Novais 2020Bandeira MDSF & Novais JS (2020) Melissopalynological characterization of honeys from the Discovery Coast, Brazil. Palynology 44: 539-550. ). Nowadays, it is recommended that this information, along with the description of the sensory and nutraceutical characteristics of honeys, be included on the labels of honeys for commercialization purposes (Mădaş et al. 2019Mădaş MN, Mărghitaş LA, Dezmirean DS, Bobiş O, Abbas O, Danthine S, Francis F, Haubruge E & Nguyen BK (2019) Labeling regulations and quality control of honey origin: a review. Food Reviews International 36: 215-240.).

In Argentina, among the honey-producing species we can mention to Apis mellifera L. (Apidae, Apini, Moure 2012Moure JS (2012) Apini Latreille, 1802. In: Moure JS, Urban D & Melo GAR (orgs.) Catalogue of bees (Hymenoptera, Apoidea) in the Neotropical Region - online version. Available at <http://moure.cria.org.br/catalogue>. Access on 14 August 2020.
http://moure.cria.org.br/catalogue...
) and native stingless bees belonging to the Meliponini tribe (Apidae, Camargo & Pedro 2013Camargo JMF & Pedro SRM (2013) Meliponini Lepeletier, 1836. In: Moure JS, Urba D & Melo GAR (orgs.) Catalogue of bees (Hymenoptera, Apoidea) in the Neotropical Region - online version. Available at <http://moure.cria.org.br/catalogue>. Access on 14 August 2020.
http://moure.cria.org.br/catalogue...
). A. mellifera is the most widespread honey-producing species in the world and the number of hives is increasing (Potts et al. 2016Potts SG, Imperatriz-Fonseca V, Ngo HT, Aizen MA, Biesmeijer JC, Breeze TD, Dicks LV, Garibaldi LA, Hill R, Settele J & Vanbergen AJ (2016) Safeguarding pollinators and their values to human well-being. Nature 540: 220-229.) despite the fact that in some countries of the Northern Hemisphere has undergone the loss of colonies due to Colony Collapse Disorder (CCD, Vanengelsdorp et al. 2017Vanengelsdorp D, Traynor KS, Andree M, Lichtenberg EM, Chen Y, Saegerman C & Cox-Foster DL (2017) Colony Collapse Disorder (CCD) and bee age impact honey bee pathophysiology. PLoS ONE 12: e0179535.).

On the other hand, 37 species of stingless bees have been identified in Argentina; they belong to 18 genera and are distributed mainly in the ecoregions of Yungas, Chaco and Atlantic Forest (Álvarez 2016Álvarez LJ (2016) Diversidad de las abejas nativas de la tribu Meliponini (Hymenoptera, Apidae) en Argentina. Tesis de Doctorado. Universidad Nacional de La Plata, La Plata. 288p.). Some of these species have been traditionally exploited in different regions of the country (Arenas 2003Arenas P (2003) Miel, Hámago y Larvas. In: Arenas P (ed.) Etnografía y Alimentación entre los Toba-Ñachilamole#ek y Wichí-Lhulku’tas del Chaco-Central (Argentina). Buenos Aires. Pp. 289-316.; Cebolla-Badie 2009Cebolla-Badie M (2009) Una etnografía sobre la miel en la Cultura Mbya - Guaraní. Editorial Abya-Yala, Quito. 147p.; Zamudio et al. 2010Zamudio F, Kujawska M & Hilgert NI (2010) Honey as medicinal and food resource. Comparison between polish and multiethnic settlements of the Atlantic Forest, Misiones, Argentina. The Open Complementary Medicine Journal 2: 58-73.; Flores et al. 2018Flores FF, Hilgert NI & Lupo LC (2018) Melliferous insects and the uses assigned to their products in the northern Yungas of Salta, Argentina. Journal of Ethnobiology and Ethnomedicine 14: 27.). For example, Tetragonisca fiebrigi (Schwarz) is an eusocial insect used by local communities; stores honey and pollen in permanent colonies generally located within pre-existing cavities, like tree holes (Michener 2013Michener CD (2013) The Meliponini. In: Vit P, Pedro SRM & Roubik DW (eds.) Pot honey. A legacy of stingless bees. Springer Verlag, New York. Pp. 3-17. ). Recently, their honeys have been included in the Argentine Food Code (<https://www.boletinoficial.gob.ar/detalleAviso/primera/206764/20190502>), a fundamental step that adds value to this product and ensures adequate mechanisms for quality control and fair trade.

In Argentina, the food sources of A. mellifera have been frequently studied (Salgado & Pire 1998Salgado CR & Pire SM (1998) Análisis polínico de las mieles del noroeste de la Provincia de Corrientes (Argentina). Darwiniana 36: 87-93.; Tamame & Naab 2003Tamame MA & Naab OA (2003) Mieles monoflorales pampeanas de Condalia microphylla Cav. y Centaurea solstitialis L.: análisis melisopalinológicos relacionados con caracteres fisicoquímicos. Revista del Museo Argentino de Ciencias Naturales 5: 371-381.; Fagúndez & Caccavari 2006Fagúndez GA & Caccavari MA (2006) Pollen analysis of honeys from the central zone of the Argentine province of Entre Ríos. Grana 45: 305-320.; Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.; Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.; Reyes et al. 2019Reyes NJF, Asesor PN, Albarracín VN, García ME & Espeche ML (2019) Caracterización palinológica de la miel de un sector de la región chaqueña de la provincia de Tucumán (Argentina). Boletín de la Sociedad Argentina de Botánica 54: 367-379.; among others).

Previous studies of the floral origin of A. mellifera honeys from the Argentine Atlantic Forest (Misiones province), revealed a dominance of the families Asteraceae, Anacardiaceae, Fabaceae, Myrtaceae, Rutaceae and Sapindaceae, and the abundant presence of pollen types belonging to native tree species (e.g., Anadenanthera colubrina (Vell.) Brenan, Balfourodendron riedelianum (Engl.) Engl., Cordia trichotoma (Vell.) Arráb. ex Steud., Eugenia uniflora L., Ilex paraguariensis A. St.-Hil., Parapiptadenia rigida (Benth.) Brenan, Peltophorum dubium (Spreng.) Taub., Schinus terebinthifolia Raddi, Sebastiania brasiliensis Spreng., Syagrus romanzoffiana (Cham.) Zanthoxylum petiolare A.St.-Hil. & Tul.), exotic trees (e.g., Citrus sp., Hovenia dulcis Thunb., Lagerstroemia indica L., Psidium guajava L.) and herbaceous species (e.g., Chamissoa altissima (Jacq.) Kunth, Leonurus japonicus Houtt., Sida rhombifolia L.) (Aquino et al. 2015Aquino D, Pellizer N & Miranda D (2015) Contenido polínico de mieles de Apis mellifera L. producidas en Misiones, Argentina. Revista Forestal Yvirareta 22: 8-12.; Miranda et al. 2010Miranda D, Keller HÁ, Silva F & Insaurralde C (2010) Flora apícola en Colonia Laharrague, Misiones, Argentina. Revista Forestal Yviraretá 17: 43-50., 2018).

Likewise, in the same ecoregion, studies of honeys and pollen loads from southeastern Brazil showed that the main botanical families used as resources were Anacardiaceae, Arecaceae, Asteraceae, Euphorbiaceae, Fabaceae, Myrtaceae, Solanaceae, among others. The main pollen types were Allophylus petiolulatus, Baccharis, Campomanesia guazumifolia, Citrus, Cocos nucifera, Eucalyptus, Ilex paraguariensis, Matayba elaeagnoides, Mikania, Myrcia, Vernonanthura and Syagrus romanzoffiana (Ramalho et al. 1991Ramalho M, Guibu LS, Giannini TC, Kleinert-Giovannini A & Imperatriz-Fonseca VL (1991) Characterization of some southern Brazilian honey and bee plants through pollen analysis. Journal of Apicultural Research 30: 81-86.; Alves & Santos 2018Alves RF & Santos FAR (2018) Pollen foraged by bees (Apis mellifera L.) on the Atlantic Forest of Bahia, Brazil. Palynology 43: 523-529.).

In Argentina, the floral resources used by stingless bee species have received attention only in recent years. In the north of Chaco province and the northwest of Córdoba province they were extensively studied by Vossler (Vossler et al. 2010Vossler FG, Tellería MC & Cunningham M (2010) Floral resources foraged by Geotrigona argentina (Apidae, Meliponini) in the Argentine Dry Chaco forest. Grana 49: 142-153., 2014; Vossler 2019Vossler FG (2019) Pollen diet assessment and flower association in Melipona orbignyi and recommendations on management and conservation of stingless bees in the Chaco dry forest of South America. Apidologie 50: 391-413., among others) and Geisa (Geisa 2020Geisa MG (2020) Análisis de los usos locales y la calidad de mieles de abejas nativas sin aguijón del noroeste de Córdoba, Argentina. Tesis de Doctorado. Universidad Nacional de Córdoba, Córdoba. 243p. ), respectively. The nectariferous resources used by T. fiebrigi was studied through the pollen characterization of its honeys for the first time in the Yungas (northwestern Argentina) by Flores & Sánchez (2010)Flores FF & Sánchez AC (2010) Primeros resultados de caracterización botánica de mieles de Tetragonisca angustula Latreille (Apidae, Meliponinae) criadas en la localidad Los Naranjos - Orán - Salta. Boletín de Sociedad Argentina de Botánica 45: 81-91., and in the Argentine Atlantic Forest by Fernández et al. (2015)Fernández LC, Huk LH, Paul RM & Salgado CR (2015) Recursos florísticos usados por Tetragonisca angustula Latreille en algunas localidades de la provincia de Misiones. Ameghiniana 52: 46., and a preliminary work (including two honey samples) by Miranda et al. (2018)Miranda D, Molina R, Aquino D, Pellizer N, Berdún A, Fernández L & Huk L (2018) Flora utilizada por Apis mellifera L. y Tetragonisca fiebrigi Schwarz en 5 departamentos de la zona centro-norte de la provincia de Misiones, Argentina. Revista Forestal Yviraretá 26: 38-54.. In some of these works the species is referred to as T. angustula Latreille; this species is synonym of T. fiebrigi, which is the only species of the genus present in Argentina (Álvarez 2016Álvarez LJ (2016) Diversidad de las abejas nativas de la tribu Meliponini (Hymenoptera, Apidae) en Argentina. Tesis de Doctorado. Universidad Nacional de La Plata, La Plata. 288p.). Fernández et al. (2015)Fernández LC, Huk LH, Paul RM & Salgado CR (2015) Recursos florísticos usados por Tetragonisca angustula Latreille en algunas localidades de la provincia de Misiones. Ameghiniana 52: 46. identified Apiaceae, Asteraceae, Fabaceae and Solanaceae as the main botanical families, and Hovenia dulcis, Cecropia pachystachya, Lagerstroemia indica, Ammi-type and Poaceae as the pollen types with the highest frequency of occurrence in the samples. On the other hand, Miranda et al. (2018)Miranda D, Molina R, Aquino D, Pellizer N, Berdún A, Fernández L & Huk L (2018) Flora utilizada por Apis mellifera L. y Tetragonisca fiebrigi Schwarz en 5 departamentos de la zona centro-norte de la provincia de Misiones, Argentina. Revista Forestal Yviraretá 26: 38-54. highlighted the botanical families Anacardiaceae, Apiaceae, Arecaceae and Asteraceae, and the pollen types Syagrus romanzoffiana, Trema micrantha, Cecropia pachystachya and Schinus terebinthifolia.

Therefore, because there is a local interest in knowing the floral resources used by A. mellifera and T. fiebrigi and in order to contribute to the list of main nectariferous species in northern Misiones and the ecorregion, the pollen content of their honeys was analysed and the most abundant botanical families and pollen types are discussed for each bee species, providing substantial information about their diet.

Materials and Methods

Study area

The study was conducted in different localities of General Manuel Belgrano, Iguazú, and San Pedro Departments, in the north of Misiones province (Fig. 1), belonging to the Upper Paraná Atlantic Forest (Galindo-Leal & Camara 2003Galindo-Leal C & Camara IG (2003) Atlantic Forest hotspot status: an overview. In: Galindo-Leal C & Camara IG (eds.) The Atlantic Forest of South América: biodiversity status, threats, and outlook. Island Press, Washington. Pp. 3-11.). The inhabitants of the study area belong to diverse cultural groups: Mbya-Guaraní ethnic groups, Europeans, Asians, Brazilians, and Paraguayans, as well as people from other Argentine provinces (Gallero & Krautstofl 2010Gallero MC & Krautstofl EC (2010) Proceso de poblamiento y migraciones en la Provincia de Misiones, Argentina (1881-1970). Avá 16: 245-264.; Zamudio et al. 2010Zamudio F, Kujawska M & Hilgert NI (2010) Honey as medicinal and food resource. Comparison between polish and multiethnic settlements of the Atlantic Forest, Misiones, Argentina. The Open Complementary Medicine Journal 2: 58-73.). At present, the landscape is composed of protected areas, and urban and rural areas where forestry-agro-livestock production systems of different sizes coexist (Furlan et al. 2015Furlan V, Cariola L, García D & Hilgert NI (2015) Caracterización de los sistemas agroforestales familiares y estrategias de uso del ambiente en el Bosque Atlántico Argentino. Gaia Scientia 9: 69-81.). Among the most widespread crops are “yerba mate” (Ilex paraguariensis A. St.-Hil.), tobacco (Nicotiana tabacum L.), citrus (Citrus sp.) and, to a lesser extent, corn (Zea mays L.), peanut (Arachis hypogaea L.), soybean (Glycine max (L.) Merr.), and tea (Camellia sinensis (L.) Kuntze). In addition, cassava (Manihot esculenta Crantz), sweet potato (Ipomoea batatas (L.) Lam) and fruit trees are cultivated in households, complemented with animal husbandry (Zamudio & Hilgert 2011Zamudio F & Hilgert NI (2011) Mieles y plantas en la medicina criolla del Norte de Misiones, Argentina. Bonplandia 20: 165-184.; Furlan et al. 2015Furlan V, Cariola L, García D & Hilgert NI (2015) Caracterización de los sistemas agroforestales familiares y estrategias de uso del ambiente en el Bosque Atlántico Argentino. Gaia Scientia 9: 69-81.). In this context, the bees A. mellifera and T. fiebrigi were selected, within the framework of ethnobiological studies carried out in the region and according to their importance of use by local communities (Zamudio et al. 2010Zamudio F, Kujawska M & Hilgert NI (2010) Honey as medicinal and food resource. Comparison between polish and multiethnic settlements of the Atlantic Forest, Misiones, Argentina. The Open Complementary Medicine Journal 2: 58-73.; Kujawska et al. 2012Kujawska M, Zamudio F & Hilgert NI (2012) Honey-based mixtures used in home medicine by nonindigenous population of Misiones, Argentina. Evidence-Based Complementary and Alternative Medicine 2012: 1-15.). The sample collection sites were selected according to the use of honey from both bees in the same production unit.

Figure 1
Collection sites of honeys of Apis mellifera and Tetragonisca fiebrigi.

Honey sampling

Between November 2006 and November 2008, 14 honey samples of A. mellifera and 39 honey samples of T. fiebrigi were collected from man-made hives (Tab. 1). In A. mellifera, 250 mL of honey per hive were collected using a conventional beekeeping extractor. In T. fiebrigi, 10 mL of honey per hive were extracted with a sterile syringe (the average annual production per hive is 1 L).

Table 1
Number of honey samples collected from different localities of northern Misiones.

Laboratory processing and microscopic analysis

Each honey sample was homogenized and processed according to Louveaux et al. (1978)Louveaux J, Maurizio A & Vorwhol G (1978) Methods of Melissopalynology. Bee World 59: 139-157. and the sediment was acetolyzed (Erdtman 1960Erdtman G (1960) The acetolysis method. Svensk Botanisk Tidskrift 54: 561-564.). The obtained microscope slides were deposited in the reference collections of Laboratory of Palynology of Universidad Nacional de Jujuy (PAL-JUA) and Instituto de Biología Subtropical of Universidad Nacional de Misiones (PAL-ETNO). Observations were made under Leica DM 500 optical microscope and microphotographs were taken with a Leica ICC50 digital camera. For quantification of the pollen types, at least 1,000 pollen grains were counted from each sample; they were determined and classified as predominant (more than 45% of the pollen grains counted), secondary (16–45%), important minor pollen (3–15%), and minor pollen (less than 3%). Honeys were classified as unifloral if there was a predominant pollen type, except for Citrus sp. with 10% (Louveaux et al. 1978Louveaux J, Maurizio A & Vorwhol G (1978) Methods of Melissopalynology. Bee World 59: 139-157.) and Eucalyptus sp. with 70% (Resolution 274/95; SAGPyA 1995SAGPyA - Secretaría de Agricultura, Ganadería, Pesca y Alimentación (1995) Resolución 274/95. Sistema de clasificación de miel teniendo como base su origen botánico. Boletín Oficial 28268: 2.). The frequency of occurrence (FO; Feller-Demalsy et al. 1987Feller-Demalsy M, Parent J & Strachan A (1987) Microscopic analysis of honeys from Alberta, Canada. Journal of Apicultural Research 26: 123-132.) was established as the percentage of a pollen type present in all the samples: very frequent (˃ 50%), frequent (20–50%), less frequent (10–20%) and rare (˂ 10%). Pollen types of wind-pollinated or nectarless plants were excluded from the frequency calculations because these plants do not contribute to honey production and their presence samples can distort botanical characterization (Louveaux et al. 1978Louveaux J, Maurizio A & Vorwhol G (1978) Methods of Melissopalynology. Bee World 59: 139-157.; Flores et al. 2015Flores FF, Lupo LC & Hilgert NI (2015) Recursos tróficos utilizados por Plebeia intermedia (Apidae, Meliponini) en la localidad de Baritú, Salta, Argentina. Caracterización botánica de sus mieles. Boletín de Sociedad Argentina de Botánica 50: 515-529.). Pollen types were identified using the reference collection of the study area and the existing literature (Markgraf & D’Antoni 1978Markgraf V & D’Antoni H (1978) Pollen flora of Argentina. The University of Arizona Press, Tucson. 208p. ; Pire et al. 1998Pire SM, Anzótegui LM & Cuadrado GA (1998) Flora Polínica del Nordeste Argentino 1. EUDENE-UNNE, Corrientes. 144p., 2001Pire SM, Anzótegui LM & Cuadrado GA (2001) Flora Polínica del Nordeste Argentino 2. EUDENE-UNNE, Corrientes. 173p., 2006Pire SM, Anzótegui LM & Cuadrado GA (2006) Flora Polínica del Nordeste Argentino 3. EUDENE-UNNE, Corrientes. 173p., 2013Pire SM, Anzótegui LM & Cuadrado GA (2013) Flora Polínica del Nordeste Argentino 4. EUDENE-UNNE, Corrientes. 168p.). The nomenclature of the pollen types follows the criterion of De Klerk & Joosten (2007)De Klerk P & Joosten H (2007) The difference between pollen types and plant taxa: a plea for clarity and scientific freedom. Eiszeitalter und Gegenwart/Quaternary Science Journal 56: 162-171..

Data analysis

For each sample, the following parameters were calculated:

a) Pollen type richness (S), i.e., the number of recorded pollen types;

b) Index of species importance (ISj ) = Mean relative abundance of j × Number of samples with j / Total number of samples

Where j is each pollen type;

c) Index of botanical family importance (IFi ) = Sum of mean relative abundance of species of i × Number of samples with i / Total number of samples.

Where i is the botanical family (Nates-Parra et al. 2013Nates-Parra G, Montoya PM, Chamorro FJ, Ramírez N, Giraldo C & Obregón D (2013) Origen geográfico y botánico de mieles de Apis mellifera (Apidae) en cuatro departamentos de Colombia. Acta Biológica Colombiana 18: 427-438.; Vossler 2019Vossler FG (2019) Pollen diet assessment and flower association in Melipona orbignyi and recommendations on management and conservation of stingless bees in the Chaco dry forest of South America. Apidologie 50: 391-413.). The proposed indexes were calculated using the Microsoft Excel program.

For the pollen diagram, data analysis and dendrogram the TILIA 1.7.16 software package was used (Grimm 2011Grimm E (2011) Tilia software. Versión 2.1.1. Illinois State Museum, Research & Collection Center, Springfield. Available at <https://www.tiliait.com/download/>. Access on 28 August 2021.
https://www.tiliait.com/download/...
). Standardised coefficients of dissimilarity were applied using the Edwards & Cavalli-Sforza coefficient and non-constrained (without sample ordination). Groups were organized according to abundance and frequency of their pollen types. Pollen types in class frequency of less than 3% were not included, because these types are present throughout the pollen spectrum and do not contribute to the differentiation of the groupings. In this way, sample groupings were established presenting affinity among them.

Results

Botanical origin and pollen richness of samples

A total of 117 pollen types were found; of them, 76 types were present in the A. mellifera samples, 104 in the T. fiebrigi samples, and 63 in the samples of both species. In A. mellifera, one pollen type was identified at division (Liliopsida), 14 at family, 22 at genus, and 38 at species levels, and one was undetermined. Richness varied between 10 and 34 pollen types, with an average of 20.5 ± 7.7. In the honeys of T. fiebrigi, one pollen type was identified at botanical division (Liliopsida), 18 at family, 30 at genus, and 54 at species levels, and one was undetermined. Richness per sample varied between 13 and 43 pollen types, with an average of 24.8 ± 7.1 (Tab. 2; see also Appendix S1 and S2, available on supplementary material <https://doi.org/10.6084/m9.figshare.16689481.v1).

Table 2
Pollen types identified in honey samples of Apis mellifera and Tetragonisca fiebrigi, and their frequency classes and frequency of occurrence in northern Misiones.

In the total of samples 14 pollen types of wind-pollinated or nectarless plants were found. The pollen types more frequents in the honeys of T. fiebrigi were Trema micrantha, Cecropia pachystachya, Celtis iguanaea, Moraceae and Piper, and in the honeys of A. mellifera were C. pachystachya, Solanaceae-type, C. iguanaea and Lycium morongii-type (Tab. 2).

Frequency classes of pollen types and classification of honeys

In honeys of A. mellifera, 3% of the pollen types were classified as predominant (present in 7 -i.e. 50%- samples), 6% as secondary, 27% as important minor, and 64% as minor pollen. In this bee species, 50% (7 samples) of the honeys were unifloral. In honeys of T. fiebrigi, 3% of the pollen types were classified as predominant (present in 23 -i.e. 59%- samples), 7% as secondary, 18% as important minor, and 72% as minor pollen. In this bee species, 59% of the honeys (23 samples) were classified as unifloral (Fig. 2).

Figure 2
Predominant pollen types in unifloral honeys of Apis mellifera and Tetragonisca fiebrigi in Misiones, Argentina.

Index of species and botanical family importance

According to the index of species importance, the pollen type Euphorbiaceae were the most abundant and frequent in the samples of A. mellifera followed for Euterpe edulis, Holocalyx balansae, Calyptranthes concinna-type, Actinostemon, Salix, Ruprechtia laxiflora, Myrcianthes pungens-type, Thinouia mucronata, Allophylus edulis, Ilex, Syagrus romanzoffiana, Gouania latifolia-type, Parapiptadenia rigida, and Baccharis-type (Tab. 3; Figs. 3; 4). On the other hand, in the samples of T. fiebrigi the pollen type most important was S. romanzoffiana followed for Schinus weinmannifolius-type, Baccharis-type, H. balansae, E. edulis, Rhamnaceae, Citrus, Leonurus japonicus, Gouania latifolia-type, A. edulis, Gomphrena perennis-type, Pouteria gardneriana, P. rigida, Zanthoxylum, and Actinostemon (Tab. 4; Figs. 3; 4).

Figure 3
Index of species importance (ISj). Only pollen types with an index equal to or higher than 2% are represented.
Figure 4
a-y. Main pollen types identified in honey samples of Apis mellifera and Tetragonisca fiebrigi – a-b. Syagrus romanzoffiana (Arecaceae); c-d. Euterpe edulis (Arecaceae); e. Parapiptadenia rigida (Fabaceae); f. Thinouia mucronata (Sapindaceae). g. Citrus (Rutaceae). h. Pouteria gardneriana (Sapotaceae). i. Actinostemon (Euphorbiaceae); j. Leonurus japonicus (Lamiaceae); k. Schinus weinmannifolius-type (Anacardiaceae); l-m. Rhamnaceae; n-o. Zanthoxylum (Rutaceae); p-q. Calyptranthes concinna-type (Myrtaceae); r-s. Holocalyx balansae (Fabaceae); t. Gomphrena perennis-type (Amaranthaceae); u. Baccharis-type (Asteraceae); v-w. Euphorbiaceae; x. Allophylus edulis (Sapindaceae); y. Ilex (Aquifoliaceae). Scale bar = 20 μm.
Table 3
Pollen types of nectariferous plants in honey samples of Apis mellifera in northern Misiones.
Table 4
Pollen types of nectariferous plants in honey samples of Tetragonisca fiebrigi in northern Misiones.

In the samples of A. mellifera, pollen types belonging to 34 botanical families were recorded, with Fabaceae presenting the highest pollen richness. However, according to the index of family importance, Fabaceae was the third most important (15%), after Euphorbiaceae (21%) and Arecaceae (17%). On the other hand, the pollen types of T. fiebrigi honeys belonged to 44 botanical families, with Fabaceae also exhibiting the highest pollen richness. Likewise, when considering the index of family importance, Fabaceae also was third, after Arecaceae (27%) and Anacardiaceae (17%) (Fig. 5).

Figure 5
Index of botanical family importance (IFi). Families with an index equal to or higher than 1% are represented in the graph.

Pollen diagram

Two main groups were distinguished in the pollen diagram prepared for A. mellifera honeys (Fig. 6). The first of them (I) concentrates samples collected in September, October and November 2007 (samples A12 to A8). These are characterized by a high presence of Holocalyx balansae, followed by the pollen types Calyptranthes concinna-type, Euphorbiaceae, Euterpe edulis, Actinostemon, Ruprechtia laxiflora, Myrcianthes pungens-type, Allophylus edulis, Ilex, Leonurus japonicus, Gouania latifolia-type, and Matayba elaeagnoides.

Figure 6
Pollen diagram of honey samples of Apis mellifera of northern Misiones.

Group II gathers samples from December 2007 (A11 and A10) and January 2008 (A11 to A14), where Euterpe edulis stood out for its high presence in the samples, accompanied by Calyptranthes concinna-type and Thinouia mucronata and to a lesser extent by Salix, Syagrus romanzoffiana, Parapiptadenia rigida and Euphorbiaceae.

On the other hand, three main groups were distinguished in the T. fiebrigi honey samples (Fig. 7, groups III, IV and V). In the first of them, the samples from 2006 (Y1, November) and 2007 (Y2 and Y3, January) presented a high abundance of the arboreal pollen type Syagrus romanzoffiana accompanied by Leonurus japonicus, Baccharis-type, Bignoniaceae and Rorippa hilariana-type. In the same group and with samples corresponding to several months of the year 2007 (samples Y10 to Y35) a high presence of the pollen type Holocalyx balansae was observed, accompanied to a lesser extent by the pollen types Schinus weinmannifolius-type, S. romanzoffiana, L. japonicus, Pouteria fragrans, P. gardneriana, Parapiptadenia rigida, Citrus, Gouania latifolia-type, Gomphrena perennis-type, Zanthoxylum, Rhamnaceae, Bignoniaceae, Euterpe edulis, Balfourodendron riedelianum-type, Aspidosperma, Anacardiaceae, Allophylus edulis and Actinostemon.

Figure 7
Pollen diagram of honey samples of Tetragonisca fiebrigi of northern Misiones.

In group IV, with samples from January, March and November 2008 (samples Y19 to Y37), a high presence of the pollen types S. romanzoffiana, Schinus weinmannifolius-type, Baccharis-type and to a lesser extent stand out Rhamnaceae, H. balansae, A. edulis, Citrus, and E. edulis were observed. While group V, with samples from November 2008 (Y38 and Y39), E. edulis stood out, followed for P. gardneriana, P. rigida, S. weinmannifolius-type and Euphorbiaceae.

Discussion

The pollen richness observed in the honey samples reflects the polylectic foraging habit of the studied bee species. These results agree with findings of studies conducted in Argentina for A. mellifera (Fagúndez & Caccavari 2006Fagúndez GA & Caccavari MA (2006) Pollen analysis of honeys from the central zone of the Argentine province of Entre Ríos. Grana 45: 305-320.; Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.; Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.; Reyes et al. 2019Reyes NJF, Asesor PN, Albarracín VN, García ME & Espeche ML (2019) Caracterización palinológica de la miel de un sector de la región chaqueña de la provincia de Tucumán (Argentina). Boletín de la Sociedad Argentina de Botánica 54: 367-379.) and T. fiebrigi (Flores & Sánchez 2010Flores FF & Sánchez AC (2010) Primeros resultados de caracterización botánica de mieles de Tetragonisca angustula Latreille (Apidae, Meliponinae) criadas en la localidad Los Naranjos - Orán - Salta. Boletín de Sociedad Argentina de Botánica 45: 81-91.; Vossler et al. 2014Vossler FG, Fagúndez GA & Blettler DC (2014) Variability of food stores of Tetragonisca fiebrigi (Schwarz) (Hymenoptera: Apidae: Meliponini) from the Argentine Chaco based on pollen analysis. Sociobiology 61: 449-460.; Fernández et al. 2015Fernández LC, Huk LH, Paul RM & Salgado CR (2015) Recursos florísticos usados por Tetragonisca angustula Latreille en algunas localidades de la provincia de Misiones. Ameghiniana 52: 46.). This behaviour was also observed in other members of the genus Tetragonisca, such as T. angustula in Bolivia (Saravia-Nava et al. 2018Saravia-Nava A, Niemeyer HM & Pinto CF (2018) Pollen types used by the native Stingless bee, Tetragonisca angustula (Latreille), in an Amazon-Chiquitano transitional forest of Bolivia. Neotropical Entomology 47: 798-807.), Brazil (Iwama & Melhem 1979Iwama S & Melhem TS (1979) The pollen spectrum of the honey of Tetragonisca angustula angustula Latreille (Apidae, Meliponinae). Apidologie 10: 275-295.; Novais et al. 2013Novais JS, Absy ML & Santos FAR (2013) Pollen grains in honeys produced by Tetragonisca angustula (Latreille, 1811) (Hymenoptera: Apidae) in tropical semi-arid areas of north-eastern Brazil. Arthropod-Plant Interactions 7: 619-632.), Colombia (Obregón et al. 2013Obregón D, Rodríguez C, Chamorro FJ & Nates-Parra G (2013) Botanical origin of Pot-Honey from Tetragonisca angustula Latreille in Colombia. In: Vit P, Pedro SRM & Roubik DW (eds.) Pot Honey. A legacy of stingless bees. Springer Verlag, New York. Pp. 337-346.), and Mexico (Martínez-Hernández et al. 1994Martínez-Hernández E, Cuadriello-Aguilar JI, Ramirez-Arriaga E, Medina-Camacho M, Sosa-Nájera MS & Melchor-Sánchez JE (1994) Foraging of Nannotrigona testaceicornis, Trigona (Tetragonisca) angustula, Scaptotrigona mexicana and Plebeia sp. in the Tacaná region, Chiapas, Mexico. Grana 33: 205-217.). In turn, it is a foraging mode commonly observed in stingless bees and A. mellifera of other Neotropical regions (Ramalho et al. 1990Ramalho M, Kleinert-Giovannini A & Imperatriz-Fonseca VL (1990) Important bee plants for stingless bees (Melipona and Trigonini) and Africanized honeybees (Apis mellifera) in neotropical habitats: a review. Apidologie 21: 469-488., 1991).

Arecaceae was the most abundant family for T. fiebrigi and the second most abundant for A. mellifera, especially with the pollen types of Syagrus romanzoffiana and Euterpe edulis. Similarly, the relevance of S. romanzoffiana was indicated for the west and centre of Misiones province (Aquino et al. 2015Aquino D, Pellizer N & Miranda D (2015) Contenido polínico de mieles de Apis mellifera L. producidas en Misiones, Argentina. Revista Forestal Yvirareta 22: 8-12.) and the states of Rio Grande do Sul and São Paulo (southern Brazil) (Ramalho et al. 1991Ramalho M, Guibu LS, Giannini TC, Kleinert-Giovannini A & Imperatriz-Fonseca VL (1991) Characterization of some southern Brazilian honey and bee plants through pollen analysis. Journal of Apicultural Research 30: 81-86.; Hilgert-Moreira et al. 2014Hilgert-Moreira SB, Nascher CA, Callegari-Jacques SM & Blochtein B (2014) Pollen resources and trophic niche breadth of Apis mellifera and Melipona obscurior (Hymenoptera, Apidae) in a subtropical climate in the Atlantic rain forest of southern Brazil. Apidologie 45: 129-141.), and of E. edulis for the states of Bahia and São Paulo, Brazil (Barroso et al. 2010Barroso RM, Reis A & Hanazaki N (2010) Etnoecologia e etnobotânica da palmeira juçara (Euterpe edulis Martius) em comunidades quilombolas do Vale do Ribeira, São Paulo. Acta Botánica Brasílica 24: 518-528.; Bandeira & Novais 2020). Likewise, the high presence of Euterpe/Syagrus pollen type in honeys of A. mellifera was highlighted from Porto Velho, São Paulo (Bosco & da Luz 2018Bosco LB & Luz CFP (2018) Pollen analysis of Atlantic forest honey from the Vale do Ribeira Region, state of São Paulo, Brazil. Grana 57: 144-157.).

In general, the role of palm trees in supplying resources to bees is associated with entomophilia, the most widespread pollination syndrome of this angiosperms family (Henderson 1986Henderson A (1986) A review of pollination studies in the Palmae. The Botanical Review 52: 221-259.; Barfod et al. 2011Barfod AS, Hagen M & Borchsenius F (2011) Twenty-five years of progress in understanding pollination mechanisms in palms (Arecaceae). Annals of Botany 108: 1503-1516.). Likewise, the predominance of the family was also documented in studies on honeys, corbicular loads and pollen pots of A. mellifera, and stingless bees from Brazil (Oliveira et al. 2009Oliveira FPM, Absy ML & Miranda IS (2009) Recurso polínico coletado por abelhas sem ferrão (Apidae, Meliponinae) em um fragmento de floresta na região de Manaus, Amazonas. Acta Amazonica 39: 505-518. ; Rech & Absy 2011Rech AR & Absy ML (2011) Pollen sources used by species of Meliponini (Hymenoptera: Apidae) along the Rio Negro channel in Amazonas, Brazil. Grana 50: 150-161.; Freitas & Novais 2014Freitas WAT & Novais JS (2014) Melissopalynology in the Brazilian Amazon: a databank of pollen types cited in the literature. Boletín de la Asociación Latinoamericana de Paleobotánica y Palinología 14: 103-136.; Hilgert-Moreira et al. 2014Hilgert-Moreira SB, Nascher CA, Callegari-Jacques SM & Blochtein B (2014) Pollen resources and trophic niche breadth of Apis mellifera and Melipona obscurior (Hymenoptera, Apidae) in a subtropical climate in the Atlantic rain forest of southern Brazil. Apidologie 45: 129-141.; Bandeira & Novais 2020). Other palm trees, such as Copernicia alba Morong and Trithrinax schizophylla Drude, are important nectar and pollen resources for A. mellifera and T. fiebrigi in the Chaco ecoregion of Argentina (Vossler et al. 2014Vossler FG, Fagúndez GA & Blettler DC (2014) Variability of food stores of Tetragonisca fiebrigi (Schwarz) (Hymenoptera: Apidae: Meliponini) from the Argentine Chaco based on pollen analysis. Sociobiology 61: 449-460.; Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.).

The presence of Syagrus romanzoffiana and Euterpe edulis, key elements of the Upper Paraná Atlantic Forest, is probably associated with the abundance of these trees in areas surrounding family production units and the extended flowering period of these species. Approximately 5 months in E. edulis (Silva & Reis 2018Silva JZD & Reis MSD (2018) Reproductive phenology and production of fruits in Euterpe edulis (Martius). Ciencia Florestal 28: 295-309.) and 9 months in S. romanzoffiana (Garcia & Barbedo 2016Garcia VA & Barbedo CJ (2016) Estudo fenológico de Bactris gasipaes Kunth, Euterpe edulis Mart. e Syagrus romanzoffiana (Cham.) Glassman no Vale do Ribeira, SP, Brasil. Hoehnea 43: 135-149.). In particular, E. edulis is a profitable non-timber forest resource from which “palmito” or “cogollo” and fruit pulp -Jejy’a- are obtained for food purposes, and its seeds for ornamental purposes (Chediack 2008Chediack SE (2008) Efecto de la explotación forestal sobre la estructura, diversidad y composición florística de los palmitales de la Selva Atlántica en Misiones, Argentina. Revista de Biología Tropical 56: 721-738.; Campanello et al. 2019Campanello PI, Bellow JV, Hilgert NI, Cockle KL, Villagra M, Francescantonio DD, García DS, Jaramillo M, Gauto OA & Goldstein G (2019) ¿Es posible el uso sostenible del bosque en Misiones? Necesidades de manejo a diferentes escalas, investigación, intervenciones de alto impacto y más recursos económicos. Ecología Austral 29: 122-137.; Villagra et al. 2019Villagra PE, Hilgert NI, García D, Álvarez JÁ, Chamorro M & Marino G (2019) Conservación por el uso en nuestros bosques nativos: ¿una utopía teórica o una oportunidad productiva? Available at <https://www.argentinaforestal.com/2019/12/17/conservacion-por-el-uso-en-nuestros-bosques-nativos-una-utopia-teorica-o-una-oportunidad-productiva/?fbclid=IwAR2uD75rEFwPK_k6KFHNAW-KfPR6eML8n-NTm-DTGndc4L7RQR5fyLluihY>. Access on 14 August 2020.
https://www.argentinaforestal.com/2019/1...
).

Fabaceae was abundant in the honeys of both species, as indicated by the richness of pollen types and the index of botanical family importance. This high presence could be linked to the representativeness of the family in the province of Misiones (Zanotti et al. 2020Zanotti CA, Keller HA & Zuloaga FO (2020) Biodiversidad de la flora vascular de la provincia de Misiones, Región Paranaense Argentina. Darwiniana nueva serie 8: 42-291.) and in the Neotropics (Ramalho et al. 1990Ramalho M, Kleinert-Giovannini A & Imperatriz-Fonseca VL (1990) Important bee plants for stingless bees (Melipona and Trigonini) and Africanized honeybees (Apis mellifera) in neotropical habitats: a review. Apidologie 21: 469-488.). The predominance of Fabaceae was also observed in studies of A. mellifera conducted in other departments of Misiones (Miranda et al. 2018Miranda D, Molina R, Aquino D, Pellizer N, Berdún A, Fernández L & Huk L (2018) Flora utilizada por Apis mellifera L. y Tetragonisca fiebrigi Schwarz en 5 departamentos de la zona centro-norte de la provincia de Misiones, Argentina. Revista Forestal Yviraretá 26: 38-54.) and other areas of Argentina (Fagúndez & Caccavari 2006Fagúndez GA & Caccavari MA (2006) Pollen analysis of honeys from the central zone of the Argentine province of Entre Ríos. Grana 45: 305-320.; Malacalza et al. 2007Malacalza NH, Mouteira MC, Baldi B & Lupano CE (2007) Characterisation of honey from different regions of the Province of Buenos Aires, Argentina. Journal of Apicultural Research 46: 8-14.; Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.; Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.; Reyes et al. 2019Reyes NJF, Asesor PN, Albarracín VN, García ME & Espeche ML (2019) Caracterización palinológica de la miel de un sector de la región chaqueña de la provincia de Tucumán (Argentina). Boletín de la Sociedad Argentina de Botánica 54: 367-379.). In northwestern Argentina, this family was relevant in honeys of T. fiebrigi (Flores & Sánchez 2010Flores FF & Sánchez AC (2010) Primeros resultados de caracterización botánica de mieles de Tetragonisca angustula Latreille (Apidae, Meliponinae) criadas en la localidad Los Naranjos - Orán - Salta. Boletín de Sociedad Argentina de Botánica 45: 81-91.) and Plebeia intermedia Wille (Flores et al. 2015Flores FF, Lupo LC & Hilgert NI (2015) Recursos tróficos utilizados por Plebeia intermedia (Apidae, Meliponini) en la localidad de Baritú, Salta, Argentina. Caracterización botánica de sus mieles. Boletín de Sociedad Argentina de Botánica 50: 515-529.). In the Neotropics, Fabaceae is important for A. mellifera (Alves & Santos 2018Alves RF & Santos FAR (2018) Pollen foraged by bees (Apis mellifera L.) on the Atlantic Forest of Bahia, Brazil. Palynology 43: 523-529.) and Melipona scutellaris Latreille in Brazil (Carvalho et al. 2001Carvalho CAL, Moreti AC, Marchini LC, Alves RM & Oliveira PCF (2001) Pollen spectrum of honey of “Uruçu” bee (Melipona scutellaris Latreille, 1811). Revista Brasileira de Biología 61: 63-67.), and for T. angustula in Bolivia (Saravia-Nava et al. 2018Saravia-Nava A, Niemeyer HM & Pinto CF (2018) Pollen types used by the native Stingless bee, Tetragonisca angustula (Latreille), in an Amazon-Chiquitano transitional forest of Bolivia. Neotropical Entomology 47: 798-807.), different localities of Brazil (Cortopassi-Laurino & Gelli 1991Cortopassi-Laurino M & Gelli DS (1991) Analyse pollinique, propriétés physico-chimiques et action antibactérienne des miels d’abeilles africanisées Apis mellifera et de Méliponinés du Brésil. Apidologie 22: 61-73.; Carvalho et al. 1999Carvalho CAL, Marchini LC & Ros PB (1999) Fontes de pólen utilizadas por Apis mellifera L. e algumas espécies de Trigonini (Apidae) em Piracicaba (SP). Bragantia 58: 49-56.; Barth et al. 2013Barth OM, Freitas AS, Sousa GL & Almeida-Muradian LB (2013) Pollen and physicochemical analysis of Apis and Tetragonisca (Apidae) honey. Interciencia 38: 280-285.; Freitas & Novais 2014Freitas WAT & Novais JS (2014) Melissopalynology in the Brazilian Amazon: a databank of pollen types cited in the literature. Boletín de la Asociación Latinoamericana de Paleobotánica y Palinología 14: 103-136.; Novais et al. 2013Novais JS, Absy ML & Santos FAR (2013) Pollen grains in honeys produced by Tetragonisca angustula (Latreille, 1811) (Hymenoptera: Apidae) in tropical semi-arid areas of north-eastern Brazil. Arthropod-Plant Interactions 7: 619-632., 2015), and southern Mexico (Martínez-Hernández et al. 1994Martínez-Hernández E, Cuadriello-Aguilar JI, Ramirez-Arriaga E, Medina-Camacho M, Sosa-Nájera MS & Melchor-Sánchez JE (1994) Foraging of Nannotrigona testaceicornis, Trigona (Tetragonisca) angustula, Scaptotrigona mexicana and Plebeia sp. in the Tacaná region, Chiapas, Mexico. Grana 33: 205-217.).

Holocalyx balansae is a frequent tree of the medium stratum of the Atlantic Forest (Martínez-Crovetto 1963Martínez-Crovetto R (1963) Esquema fitogeográfico de la provincia de Misiones. Bonplandia 1: 171-223.) that provides important services to the local fauna; indeed, the natural hollows of this tree are used as nesting sites by stingless bees (Colleselli & Gómez-Olivera 2008Colleselli LMD & Gómez-Olivera NN (2008) Las yateí (Tetragonisca angustula) en el Refugio Privado de Vida Silvestre Yacutinga, Misiones, Argentina. Tesis de Tecnicatura Universitaria. Universidad Nacional de Misiones, Misiones. 52p.). In this study, its importance as a nectar resource for the studied bees was recorded.

Parapiptadenia rigida, with a frequency of occurrence higher than 60% in honey samples, is a conspicuous tree in Upper Paraná Atlantic Forest (Martínez-Crovetto 1963Martínez-Crovetto R (1963) Esquema fitogeográfico de la provincia de Misiones. Bonplandia 1: 171-223.) and intensely visited by A. mellifera in the northwest of Misiones (Miranda et al. 2010Miranda D, Keller HÁ, Silva F & Insaurralde C (2010) Flora apícola en Colonia Laharrague, Misiones, Argentina. Revista Forestal Yviraretá 17: 43-50.). The present results agree with findings reported for honeys of A. mellifera from other areas of Misiones (Aquino et al. 2015Aquino D, Pellizer N & Miranda D (2015) Contenido polínico de mieles de Apis mellifera L. producidas en Misiones, Argentina. Revista Forestal Yvirareta 22: 8-12.) and for honeys of Scaptotrigona jujuyensis (Basilio et al. 2013Basilio AM, Spagarino C, Landi L & Achával B (2013) Miel de Scaptotrigona jujuyensis en dos localidades de Formosa, Argentina. In: Vit P & Roubik DW (eds.) Stingless bees process honey and pollen in cerumen pots. Universidad de Los Andes, Mérida. Pp. 1-8.) from Formosa province. In addition, the botanical genus includes other species of melliferous importance, such as P. excelsa (Griseb.) Burkart for stingless bees (Flores & Sánchez 2010Flores FF & Sánchez AC (2010) Primeros resultados de caracterización botánica de mieles de Tetragonisca angustula Latreille (Apidae, Meliponinae) criadas en la localidad Los Naranjos - Orán - Salta. Boletín de Sociedad Argentina de Botánica 45: 81-91.; Flores et al. 2015Flores FF, Lupo LC & Hilgert NI (2015) Recursos tróficos utilizados por Plebeia intermedia (Apidae, Meliponini) en la localidad de Baritú, Salta, Argentina. Caracterización botánica de sus mieles. Boletín de Sociedad Argentina de Botánica 50: 515-529.) and A. mellifera in northwestern Argentina (Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.); and P. zehntneri (Harms) M. P. Lima & H. Lima C., a resource visited by A. mellifera and stingless bees from northeastern Brazil (Santos et al. 2006Santos FAR, Oliveira JM, Oliveira PP, Leite KRB & Carneiro CE (2006) Plantas do Semi-Árido importantes para as abelhas. In: Santos FAR (ed.) Apium plantae. IMSEAR, Recife. Pp. 61-86.).

The pollen characteristics observed for Schinus weinmannifolius-type suggest that it may correspond to Astronium fraxinifolium Schott, Schinus terebinthifolius Raddi or S. weinmannifolius Engl., three species present in the province of Misiones (Zanotti et al. 2020Zanotti CA, Keller HA & Zuloaga FO (2020) Biodiversidad de la flora vascular de la provincia de Misiones, Región Paranaense Argentina. Darwiniana nueva serie 8: 42-291.). Indeed, the family Anacardiaceae includes species that are essential nectar sources for bees in other areas of Argentina, such as Schinopsis balansae Engl., S. lorentzii (Griseb.) Engl. and Schinus fasciculatus (Griseb.) I.M. Johnst. in the Chaco province (Vossler et al. 2014Vossler FG, Fagúndez GA & Blettler DC (2014) Variability of food stores of Tetragonisca fiebrigi (Schwarz) (Hymenoptera: Apidae: Meliponini) from the Argentine Chaco based on pollen analysis. Sociobiology 61: 449-460.; Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.), Astronium balansae Engl. in Corrientes (Salgado & Pire 1998Salgado CR & Pire SM (1998) Análisis polínico de las mieles del noroeste de la Provincia de Corrientes (Argentina). Darwiniana 36: 87-93.), and Schinus areira L. and other Schinus and Schinopsis species in northwest of Argentina (Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.; Reyes et al. 2019Reyes NJF, Asesor PN, Albarracín VN, García ME & Espeche ML (2019) Caracterización palinológica de la miel de un sector de la región chaqueña de la provincia de Tucumán (Argentina). Boletín de la Sociedad Argentina de Botánica 54: 367-379.).

The Euphorbiaceae pollen type was found with high presence in the honeys of A. mellifera; those pollen types should be studied in depth to detect the plants that provide nectar, as Actinostemon in the honeys of both bee species. Other important Euphorbiaceae in the north of Argentina are Cnidoscolus loasoides (Pax) I. M. Johnst., Croton argenteus L., Croton bonplandianus Baill., Croton lachnostachyus Baill., Jatropha spp., Sapium haematospermum Müll. Arg. and Sebastiania brasiliensis Spreng. for T. fiebrigi (Flores & Sánchez 2010Flores FF & Sánchez AC (2010) Primeros resultados de caracterización botánica de mieles de Tetragonisca angustula Latreille (Apidae, Meliponinae) criadas en la localidad Los Naranjos - Orán - Salta. Boletín de Sociedad Argentina de Botánica 45: 81-91.; Vossler et al. 2014Vossler FG, Fagúndez GA & Blettler DC (2014) Variability of food stores of Tetragonisca fiebrigi (Schwarz) (Hymenoptera: Apidae: Meliponini) from the Argentine Chaco based on pollen analysis. Sociobiology 61: 449-460.) and A. mellifera (Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.).

The Baccharis pollen type involves species of the genus Baccharis, which are abundant nectar resources in Neotropical areas (Ramalho et al. 1990Ramalho M, Kleinert-Giovannini A & Imperatriz-Fonseca VL (1990) Important bee plants for stingless bees (Melipona and Trigonini) and Africanized honeybees (Apis mellifera) in neotropical habitats: a review. Apidologie 21: 469-488., 1991). In the north of Argentina (Salta and Chaco provinces), Baccharis species are sources of nectar for the stingles bee T. fiebrigi and Geotrigona argentina Camargo & Moure (Flores & Sánchez 2010Flores FF & Sánchez AC (2010) Primeros resultados de caracterización botánica de mieles de Tetragonisca angustula Latreille (Apidae, Meliponinae) criadas en la localidad Los Naranjos - Orán - Salta. Boletín de Sociedad Argentina de Botánica 45: 81-91.; Vossler et al. 2010Vossler FG, Tellería MC & Cunningham M (2010) Floral resources foraged by Geotrigona argentina (Apidae, Meliponini) in the Argentine Dry Chaco forest. Grana 49: 142-153., 2014), and for A. mellifera in different regions of country (Salgado & Pire 1998Salgado CR & Pire SM (1998) Análisis polínico de las mieles del noroeste de la Provincia de Corrientes (Argentina). Darwiniana 36: 87-93.; Fagúndez & Caccavari 2006Fagúndez GA & Caccavari MA (2006) Pollen analysis of honeys from the central zone of the Argentine province of Entre Ríos. Grana 45: 305-320.; Costas et al. 2013Costas MC, Vergara-Roig VA & Kivatinitz SC (2013) A melissopalynological study of artisanal honey produced in Catamarca (Argentina). Grana 52: 229-237.; Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.; Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.). Particularly in the northwest of Misiones, Baccharis-type is frequent (37%) in the honeys of A. mellifera (Aquino et al. 2015Aquino D, Pellizer N & Miranda D (2015) Contenido polínico de mieles de Apis mellifera L. producidas en Misiones, Argentina. Revista Forestal Yvirareta 22: 8-12.) and some species of Baccharis (Baccharis oxycodonta DC., B. tandilensis Speg.) are visited by A. mellifera during the summer (Miranda et al. 2010Miranda D, Keller HÁ, Silva F & Insaurralde C (2010) Flora apícola en Colonia Laharrague, Misiones, Argentina. Revista Forestal Yviraretá 17: 43-50.).

Other species abundantly represented in this study were Pouteria gardneriana, Ruprechtia laxiflora, Thinouia mucronata, and pollen types of the family Myrtaceae (Calyptranthes concinna-type, Myrcianthes pungens-type); this is the first record of these species in palynological studies in Argentina. The Myrtaceae family includes important nectar species for bees, as highlighted by Ramalho et al. (1990)Ramalho M, Kleinert-Giovannini A & Imperatriz-Fonseca VL (1990) Important bee plants for stingless bees (Melipona and Trigonini) and Africanized honeybees (Apis mellifera) in neotropical habitats: a review. Apidologie 21: 469-488..

The index of importance for Allophylus edulis estimated in this study was 5.4% for both bee species. This result highlights the importance of the tree as a nectar source, as reported for honeys of T. fiebrigi (Flores & Sánchez 2010Flores FF & Sánchez AC (2010) Primeros resultados de caracterización botánica de mieles de Tetragonisca angustula Latreille (Apidae, Meliponinae) criadas en la localidad Los Naranjos - Orán - Salta. Boletín de Sociedad Argentina de Botánica 45: 81-91.), Plebeia intermedia (Flores et al. 2015Flores FF, Lupo LC & Hilgert NI (2015) Recursos tróficos utilizados por Plebeia intermedia (Apidae, Meliponini) en la localidad de Baritú, Salta, Argentina. Caracterización botánica de sus mieles. Boletín de Sociedad Argentina de Botánica 50: 515-529.), and A. mellifera from northwestern Argentina (Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.). However, it is not as relevant in other areas of the country, as observed in the honeys of A. mellifera from the Chaco province (Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.).

The index of importance of Salix was 5.3% for both bees, with this value being higher for the honeys of A. mellifera (Fig. 3). The pollen type possibly corresponds to the species Salix humboldtiana Willd., a tree associated with water sources (rivers, lagoons, estuaries), and documented as a frequent resource in honeys of A. mellifera from the Yungas (Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.) and the Chaco ecoregion of Argentina (Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.).

The Rhamnaceae pollen type was also abundant, showing a high frequency (> 50%) in the studied honeys. The members of the family are good sources of nectar for bees. For example, Condalia microphylla Cav., Discaria Americana Gillies & Hook, Sarcomphalus mistol (Griseb.) Hauenshild and Scutia buxifolia Reissek are floral resources visited for A. mellifera, G. argentina and T. fiebrigi in other ecoregions of Argentina (Tamame & Naab 2003Tamame MA & Naab OA (2003) Mieles monoflorales pampeanas de Condalia microphylla Cav. y Centaurea solstitialis L.: análisis melisopalinológicos relacionados con caracteres fisicoquímicos. Revista del Museo Argentino de Ciencias Naturales 5: 371-381.; Tellería et al. 2006Tellería MC, Salgado CR & Cuadrado AC (2006) Rhamnaceae asociadas a mieles fétidas en Argentina. Revista del Museo Argentino de Ciencias Naturales 8: 237-241.; Vossler et al. 2010Vossler FG, Tellería MC & Cunningham M (2010) Floral resources foraged by Geotrigona argentina (Apidae, Meliponini) in the Argentine Dry Chaco forest. Grana 49: 142-153., 2014; Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.; Reyes et al. 2019Reyes NJF, Asesor PN, Albarracín VN, García ME & Espeche ML (2019) Caracterización palinológica de la miel de un sector de la región chaqueña de la provincia de Tucumán (Argentina). Boletín de la Sociedad Argentina de Botánica 54: 367-379.). Likewise, Scutia/Condalia-type is the dominant pollen type in honeys of A. mellifera in northwestern Argentina (Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.). In Misiones and the state of Santa Catarina, Brazil, Hovenia dulcis Thunb. is a key resource for A. mellifera (Miranda et al. 2018Miranda D, Molina R, Aquino D, Pellizer N, Berdún A, Fernández L & Huk L (2018) Flora utilizada por Apis mellifera L. y Tetragonisca fiebrigi Schwarz en 5 departamentos de la zona centro-norte de la provincia de Misiones, Argentina. Revista Forestal Yviraretá 26: 38-54.) and T. angustula (Freitas et al. 2010Freitas AS, Barth OM & Luz CFP (2010) Análise polínica comparativa e origem botânica de amostras de mel de Meliponinae (Hymenoptera, Apidae) do Brasil e da Venezuela. Mensagem Doce 106: 2-9.). In fact, in this work a unifloral honey of Gouania latifolia-type was found; given the observed pollinic characters, it would correspond to Gouania latifolia Reissek or G. ulmifolia Hook. & Arn., two species present in Misiones province (Zanotti et al. 2020Zanotti CA, Keller HA & Zuloaga FO (2020) Biodiversidad de la flora vascular de la provincia de Misiones, Región Paranaense Argentina. Darwiniana nueva serie 8: 42-291.). In the centre of Misiones province, G. ulmifolia was found in honeys of T. fiebrigi (Miranda et al. 2018Miranda D, Molina R, Aquino D, Pellizer N, Berdún A, Fernández L & Huk L (2018) Flora utilizada por Apis mellifera L. y Tetragonisca fiebrigi Schwarz en 5 departamentos de la zona centro-norte de la provincia de Misiones, Argentina. Revista Forestal Yviraretá 26: 38-54.) and in the forests of Brazil, Gouania cf. latifolia is considered an attractive species for A. mellifera (Polatto et al. 2014Polatto LP, Chaud-Neto J & Alves-Junior VV (2014) Influence of abiotic factors and floral resource availability on daily foraging activity of bees. Journal of Insect Behavior 27: 593-612.), as Gouania lupuloides (L.) Urb. in Chiapas, Mexico for T. angustula (Martínez-Hernández et al. 1994Martínez-Hernández E, Cuadriello-Aguilar JI, Ramirez-Arriaga E, Medina-Camacho M, Sosa-Nájera MS & Melchor-Sánchez JE (1994) Foraging of Nannotrigona testaceicornis, Trigona (Tetragonisca) angustula, Scaptotrigona mexicana and Plebeia sp. in the Tacaná region, Chiapas, Mexico. Grana 33: 205-217.).

Among herbs, Leonurus japonicus had a high presence in the studied honeys. Individuals of this species grow scattered in disturbed sectors and their fragrant flowers are used by A. mellifera in the northwest of Misiones between February and April (Miranda et al. 2010Miranda D, Keller HÁ, Silva F & Insaurralde C (2010) Flora apícola en Colonia Laharrague, Misiones, Argentina. Revista Forestal Yviraretá 17: 43-50.). Studies of honeys from north Argentina reported this species as a minor resource for T. fiebrigi (Flores & Sánchez 2010Flores FF & Sánchez AC (2010) Primeros resultados de caracterización botánica de mieles de Tetragonisca angustula Latreille (Apidae, Meliponinae) criadas en la localidad Los Naranjos - Orán - Salta. Boletín de Sociedad Argentina de Botánica 45: 81-91.) and for A. mellifera (Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.).

The Citrus pollen type represents wild and cultivated species present in farms (Stampella et al. 2013Stampella PC, Lambaré DA, Hilgert NI & Pochettino ML (2013) What the Iberian conquest bequeathed to us: the fruit trees introduced in Argentine Subtropic - their history and importance in present traditional medicine. Evidence-Based Complementary and Alternative Medicine 2013: 1-17.), which are sources of nectar and pollen for bees, as confirmed for different regions (Ramalho et al. 1990Ramalho M, Kleinert-Giovannini A & Imperatriz-Fonseca VL (1990) Important bee plants for stingless bees (Melipona and Trigonini) and Africanized honeybees (Apis mellifera) in neotropical habitats: a review. Apidologie 21: 469-488., 1991; Martínez-Hernández et al 1994; Flores & Sánchez 2010Flores FF & Sánchez AC (2010) Primeros resultados de caracterización botánica de mieles de Tetragonisca angustula Latreille (Apidae, Meliponinae) criadas en la localidad Los Naranjos - Orán - Salta. Boletín de Sociedad Argentina de Botánica 45: 81-91.; Miranda et al. 2010Miranda D, Keller HÁ, Silva F & Insaurralde C (2010) Flora apícola en Colonia Laharrague, Misiones, Argentina. Revista Forestal Yviraretá 17: 43-50.; Novais et al. 2013Novais JS, Absy ML & Santos FAR (2013) Pollen grains in honeys produced by Tetragonisca angustula (Latreille, 1811) (Hymenoptera: Apidae) in tropical semi-arid areas of north-eastern Brazil. Arthropod-Plant Interactions 7: 619-632.; Aquino et al. 2015Aquino D, Pellizer N & Miranda D (2015) Contenido polínico de mieles de Apis mellifera L. producidas en Misiones, Argentina. Revista Forestal Yvirareta 22: 8-12.; Flores et al. 2015Flores FF, Lupo LC & Hilgert NI (2015) Recursos tróficos utilizados por Plebeia intermedia (Apidae, Meliponini) en la localidad de Baritú, Salta, Argentina. Caracterización botánica de sus mieles. Boletín de Sociedad Argentina de Botánica 50: 515-529.; Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.; Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.; Reyes et al. 2019Reyes NJF, Asesor PN, Albarracín VN, García ME & Espeche ML (2019) Caracterización palinológica de la miel de un sector de la región chaqueña de la provincia de Tucumán (Argentina). Boletín de la Sociedad Argentina de Botánica 54: 367-379.). Another abundant Rutaceae genus in the honeys was Zanthoxylum, which was also be observed in the honeys of A. mellifera (Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.) and Plebeia intermedia (Flores et al. 2015Flores FF, Lupo LC & Hilgert NI (2015) Recursos tróficos utilizados por Plebeia intermedia (Apidae, Meliponini) en la localidad de Baritú, Salta, Argentina. Caracterización botánica de sus mieles. Boletín de Sociedad Argentina de Botánica 50: 515-529.) from northwestern Argentina, and in corbicular and stored pollen of Tetragonisca angustula from Bolivia (Saravia-Nava et al. 2018Saravia-Nava A, Niemeyer HM & Pinto CF (2018) Pollen types used by the native Stingless bee, Tetragonisca angustula (Latreille), in an Amazon-Chiquitano transitional forest of Bolivia. Neotropical Entomology 47: 798-807.). In Argentina, the genus includes botanical species that are frequent sources of nectar for A. mellifera, such as Zanthoxylum coco Gillies ex Hook. F. & Arn. in Yungas and Chaco ecoregions (Costas et al. 2013Costas MC, Vergara-Roig VA & Kivatinitz SC (2013) A melissopalynological study of artisanal honey produced in Catamarca (Argentina). Grana 52: 229-237.; Sánchez & Lupo 2017Sánchez AC & Lupo LC (2017) Pollen analysis of honeys from the northwest of Argentina: Province of Jujuy. Grana 56: 462-474.), Z. rhoifolium Lam. in the Chaco ecoregion (Salgado-Laurenti et al. 2017Salgado-Laurenti CR, Tellería MC & Coronel JM (2017) Botanical and geographical origin of honey from the dry and humid Chaco ecoregions (Argentina). Grana 56: 450-461.), and Z. petiolare A.St.-Hil. & Tul. in the Atlantic Forest (Miranda et al. 2010Miranda D, Keller HÁ, Silva F & Insaurralde C (2010) Flora apícola en Colonia Laharrague, Misiones, Argentina. Revista Forestal Yviraretá 17: 43-50.). A particular case was that of Balfourodendron riedelianum (Engl.) Engl., which was only found in T. fiebrigi honeys (with 8% frequency of occurrence), but is considered a tree intensely visited by A. mellifera (Miranda et al. 2010Miranda D, Keller HÁ, Silva F & Insaurralde C (2010) Flora apícola en Colonia Laharrague, Misiones, Argentina. Revista Forestal Yviraretá 17: 43-50.).

Another important crop for local people, such as Ilex paraguariensis A. St.-Hil., is possibly also important for the studied bees. The presence of the pollen type Ilex and an index of importance of 2.8% in honeys of A. mellifera support our assumption and the potential nectar contribution of the species. In previous studies, Aquino et al. (2015)Aquino D, Pellizer N & Miranda D (2015) Contenido polínico de mieles de Apis mellifera L. producidas en Misiones, Argentina. Revista Forestal Yvirareta 22: 8-12. found similar results in honeys of A. mellifera from Misiones and a frequency of occurrence of 74%.

This study presents the first case of a unifloral honey of Gomphrena perennis-type, which includes pollens of species present in Misiones, such as Gomphrena celosioides Mart., G. elegans Mart., G. graminea Moq., Hebanthe eriantha (Poir.) Pedersen, Pfaffia glomerata (Spreng.) Pedersen, P. gnaphaloides (LF) Mart. and P. tuberosa (Spreng.) Hicken (Zanotti et al. 2020Zanotti CA, Keller HA & Zuloaga FO (2020) Biodiversidad de la flora vascular de la provincia de Misiones, Región Paranaense Argentina. Darwiniana nueva serie 8: 42-291.). Likewise, in Brazil the predominance of Gomphrena demissa has been reported in a honey sample of T. angustula (Novais et al. 2015Novais JS, Garcez ACA, Absy ML & Santos FAR (2015) Comparative pollen spectra of Tetragonisca angustula (Apidae, Meliponini) from the Lower Amazon (N Brazil) and caatinga (NE Brazil). Apidologie 46: 417-431.). However, it is necessary to investigate the contamination of nectar or operculated honey by pollen, since many members of the Amaranthaceae family have anemophilous pollination (Recio et al. 1998Recio M, Trigo MM, Toro FJ & Cabezudo B (1998) Incidencia del polen de Chenopodiaceae-Amaranthaceae en la atmósfera de Málaga y su relación con los parámetros meteorológicos. Acta Botánica Malacitana 23: 121-131.).

The pollen types of wind-pollinated or nectarless plants found in the samples belong to species that provide pollen to bees (Barth 1989Barth OM (1989) O pólen no mel brasileiro. Editorial Luxor, Rio de Janeiro. 150p.; Silva et al. 2014Silva CI, Imperatriz-Fonseca VL, Groppo M, Bauermann SG, Saraiva AM, Queiroz EP, Evaldt ACP,Aleixo KP, Castro JP, Castro MMN, Faria LB, Ferreira-Caliman MJ, Wolff JL, Paulino-Neto HF & Garófalo CA (2014) Catálogo polínico das plantas usadas por abelhas no campus da USP de Ribeirão Preto. Holos, Ribeirão Preto. 153p.) and their presence in nectar or honey probably is associated with the secondary and quaternary enrichment proposed by Persano-Oddo et al. (2007)Persano-Oddo L, Piana ML & Ricciardelli-D’Albore G (2007) I miele regionali italiani. Caratterizzazione melissopalinologica. Ministero delle Politiche Agricole Alimentari e Forestali. C.R.A.-Istituto Sperimentale per la Zoologia Agraria, Sezione di Apicoltura, Roma. 139p.. Likewise, several of them were registered in melisopalinological studies carried out in other localities in Misiones (Fernández et al. 2015Fernández LC, Huk LH, Paul RM & Salgado CR (2015) Recursos florísticos usados por Tetragonisca angustula Latreille en algunas localidades de la provincia de Misiones. Ameghiniana 52: 46.) and Brazil (Barth 1989Barth OM (1989) O pólen no mel brasileiro. Editorial Luxor, Rio de Janeiro. 150p.; Novais et al. 2013Novais JS, Absy ML & Santos FAR (2013) Pollen grains in honeys produced by Tetragonisca angustula (Latreille, 1811) (Hymenoptera: Apidae) in tropical semi-arid areas of north-eastern Brazil. Arthropod-Plant Interactions 7: 619-632., 2015; Freitas & Novais 2014Freitas WAT & Novais JS (2014) Melissopalynology in the Brazilian Amazon: a databank of pollen types cited in the literature. Boletín de la Asociación Latinoamericana de Paleobotánica y Palinología 14: 103-136.).

In this study, we have observed that bees use nectar resources, belonging to wild and cultivated plants. On the other hand, it was possible to establish an association of the pollen types observed depending on the temporal origin of the honey sample. Associations that will allow us to predict the pollen content of samples in a future. For example, in the samples of both bees Holocalyx balansae was a frequent pollen type in the period studied and abundant in those samples collected in the spring season, accompanied by the pollen types Actinostemon, Euterpe edulis, Leonurus japonicus, Matayba elaeagnoides and Gouania latifolia-type. Additionally, other pollen indicators in that period were Calyptranthes concinna-type, Euphorbiaceae, Ilex, Myrcianthes pungens-type, Salix and Ruprechtia laxiflora in the samples of A. mellifera and Allophylus edulis, Aspidosperma, Citrus, Parapiptadenia rigida, Pouteria fragrans, P. gardneriana, Rhamanceae and Zanthoxylum in the samples of T. fiebrigi.

Likewise, Syagrus romanzoffiana, Schinus weinmannifolius-type and Baccharis-type make up an abundant group associated with the summer period, despite being found to a lesser extent and frequently in other samples of this study. Linked to them, it was possible to observe the presence of Salix and Thinouia mucronata, as well as other pollen types reported for the spring season (A. edulis, C. concinna-type, Citrus, E. edulis, Euphorbiaceae, P. rigida and Rhamnaceae). Lastly, Gomphrena perennis-type was an abundant pollen type in the winter T. fiebrigi samples, accompanied by Actinostemon, Bowlesia, P. rigida, and Pouteria fragrans.

The list of recorded nectariferous plants is a substantial contribution for the management of apiaries and meliponaries, allowing the local producer to select which plants to grow as well as to conserve vegetation patches containing these resources.

Acknowledgements

We thank CONICET, for the Post-Graduate Scholarship granted to Fabio Flores; to the CIES, of National Parks Administration, for allowing the researchers to working their premises in Iguazú at the beginning of these studies; to the Yacutinga Private Reserve, which partially funded this work and provided logistic support in honey collection; to the Rufford Foundation, for financial support to F. Zamudio. A special thanks to the residents of the north of Misiones who kindly provided the honey samples; and the reviewers who contributed to improving the work.

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Supplementary Material

See supplementary material at <https://doi.org/10.6084/m9.figshare.16689481.v1 >

Publication Dates

  • Publication in this collection
    22 Oct 2021
  • Date of issue
    2021

History

  • Received
    22 May 2020
  • Accepted
    04 Sept 2020
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