Melipona Beecheii Research Articles

Characterization and identification of lactic acid bacteria from Mexican stingless bees (Apidae: Meliponini)

Stingless bees (family Apidae; tribe Meliponini), native from Mexico, are essential in tropical ecosystems. They are responsible for the pollination of many crops, native flora, and honey production. Lactic Acid Bacteria (LAB) are a regular host of the bee microbiota. LAB provide beneficial effects such as decreasing bacterial and parasitic pathogens infections and enhancing beehive honey production. Four different stingless bee species were sampled in the southeast of Mexico (Veracruz State) and identified as Melipona beecheii, Scaptotrigona pectoralis, Plebeia llorentei and Plebeia jatiformis. Twelve LAB strains were isolated from the bee gastrointestinal tract and characterised by microbiologic features, carbohydrates fermentation profile, antibiogram and phylogenetic reconstructions through distance and Bayesian inference methods, selecting two genes with hypervariable regions (16S rRNA and pheS). The species were characterised as gram-positive and catalase-negative as rods and cocci. Moreover, most of the species identified were able to use diverse polysaccharides as the only carbon source. Lactobacillaceae resulted in resistance to ciprofloxacin and Leuconstaceae to cotrimoxazole. Finally, ten strains could be identified by both phylogenetic reconstructions as Lactiplantibacillus plantarum (2), Weissella paramesenteroides (3), Leuconostoc citreum (2), and Apilactobacillus spp. (3). This is the first report of LAB isolated from Mexican stingless bees to the best of our knowledge.

La meliponicultura en la Reserva de la Biósfera de Los Petenes en la península de Yucatán, México. Una iniciativa decolonial

There have been multiple project initiatives in the Yucatan Peninsula aimed to rescue the melipona bee (Melipona beecheii). Through an ethnographic study in the Los Petenes biosphere reserve in Campeche, Mexico, the present study registered the revitalization of biocultural elements associated with the melipona. From a decolonial approach, the study concludes that this revitalization promotes cultural identity, and the defense of the territory, complementing conservation efforts. Finally, the study recommends the implementation of an instrument to evaluate the biocultural dimension, with the purpose of contributing to the conservation of protected natural areas.

Characterization of a non-glycosylated fraction from honey proteins of Melipona beecheii with antimicrobial activity against Escherichia coli O157:H7.

To analyse the non-glycosylated protein fraction from Melipona beecheii honey for antimicrobial activity against Escherichia coli O157:H7. The proteins from M. beecheii honey were separated according to their degree of glycosylation using Concanavalin A-affinity chromatography. The total protein extract and its fractions were analysed by 1D and 2D electrophoresis. We also determined the antimicrobial and antihaemolytic activities of the total protein extract and the non-glycosylated fraction. Furthermore, we evaluated the effect of this non-glycosylated fraction for the expression of the Stx1, Stx2, EAE and HlyA pathogen genes. Melipona beecheii honey contained at least 24 proteins with molecular weights ranging between 7·6 and 95kDa and isoelectric points between 3 and 10, three proteins from the 24 are non-glycosylated. The non-glycosylated fraction had an MIC90 of 1·128µgml-1 , and this fraction inhibited the haemolytic activity of the pathogen, as well as reduced the expression of Stx1, Stx2 and HlyA. The MbF1-2 protein from the non-glycosylated fraction was sequenced and identified as a homologue of the royal jelly-like protein of Melipona quadrifasciata. The non-glycosylated protein fraction from M. beecheii honey greatly contributes to antibacterial activity and it is composed of at least three proteins, of which MbF1-2 provided over 50% of the antimicrobial activity. The study showed significant antimicrobial activity from several proteins present in the honey of M. beecheii. Interestingly, the non-glycosylated protein fraction demonstrated antihaemolytic activity and adversely affected the expression of virulence genes in Escherichia coli O157:H7; these proteins have the potential to be used in developing therapeutic agents against this bacterium.

Indicadores bioculturales en proyectos de gestión ambiental. El caso de la meliponicultura en Yucatán

El presente trabajo revela diferentes criterios y metodologías para el ejercicio de evaluación de proyectos de gestión ambiental. Aunado a la carencia de instrumentos de evaluación que incorporen indicadores de impacto de la dimensión biocultural, se constata la tendencia a privilegiar una valoración mercantilista de proyectos, incidiendo en la gestión ambiental. Con el apoyo teórico de la valoración relacional (relational values) y a través de la categorización de códigos emergentes de datos recabados a través de métodos etnográficos, el presente trabajo concluye con una propuesta para la aplicación de indicadores para conocer la incidencia en la dimensión biocultural en el caso de proyectos para el rescate de la meliponicultura, es decir, el cultivo de la abeja nativa comúnmente conocida como abeja melipona o Xunancab (en Maya) (Melipona beecheii), en la península de Yucatán.

Floral resources collected by four native bees species in southern Mexico

Melissopalynological analysis of 14 honey samples of Tetragonisca angustula (two), Scaptotrigona mexicana (six), Melipona beecheii (three) and Melipona solani (three) from Soconusco region in Chiapas, Mexico was performed. A total of 79 taxa were identified, being Asteraceae, Euphorbiaceae, Fabaceae, Melostomataceae and Rubiaceae the predominant pollen types. Melissopalynological analysis reported a monofloral composition for Melipona beecheii honeys, where the most predominant pollen type was Fabaceae (54.2%). T. angustula, Scaptotrigona mexicana and Melipona solani honey samples were multifloral and predominant pollen types were Asteraceae, Euphorbiaceae, Fabaceae, Melastomataceae and Solanaceae. The most frequent polliniferous pollen type were Asteraceae, Bernardia interrupta, Euphorbia heterophyla and Miconia. Shannon–Wiener diversity index (H′) for Melipona beecheii 2016 (2.21) and Melipona solani 2017 (1.64) were lower meanwhile honeys of Melipona beecheii 2017, S. mexicana 2017, Melipona solani 2016 and T. angustula 2017 were considered as fairly diverse, because they visited a wider range of plant species, by preferring shrub and wild strata such as: Asteraceae, Euphorbia, Euphorbiaceae, Miconia and Mimosa. Our results showed that the foraging behaviour of these bee species is polylectic, as no taxa was found to be more than 70% of the counted grains. Since the study of the collection of meliponine flower resources in the southern region of Mexico have not been studied yet, it is not possible to analyse the foraging behaviour, the use of resources and the management of these species in this region.

Effects of phenolic and protein extracts from Melipona beecheii honey on pathogenic strains of Escherichia coli and Staphylococcus aureus.

Antimicrobial effects of Melipona beecheii honey have been attributed to diverse factors, in this sense, certain components such as proteins and phenolics could explain relevant aspects of its antimicrobial activity. The aim of this study was to evaluate the antibacterial activity of phenolic and protein extracts from M. beecheii honey against two bacterial pathogens: Staphylococcus aureus and Escherichia coli. With respect to phenolic content, HPLC analysis allowed the identification of phenolic acids like chlorogenic acid, caffeic acid, and flavonoids like catechin, myricetin, quercetin and apigenin. On the other hand, seven bands with molecular weight from 7.6 to 95kDa were detected in protein extract by SDS-PAGE system. It was determined the antibacterial activity of both extracts, with MICs lower than 145µg/mL and 60µg/mL for the phenolic and protein extracts respectively. These results indicate that phenolic and protein components of M. beecheii honey contribute significantly to the antibacterial activity.

FROM MANAGING BEES TO SCHEDULING LIFE VIA DAY SIGNS: THE LOG HIVE AS A WINDOW FOR COMMUNICATION IN PRE-HISPANIC MAYA SOCIETY

Mayas from the pre-Hispanic period documented important facts and life experiences via a writing system consisting of hieroglyphs. To timetable these events, they used time units such as the so-called day signs, whose origins are largely unknown. Classic Maya civilization included a tradition of keeping native stingless bees in horizontally oriented, hollowed-out logs for the production of honey and wax. The legacy and value of this stingless-bee keeping, also called meliponiculture, can be found in their culture and religion. The most commonly kept species was Melipona beecheii , which the Mayas knew by the name of Xunan-kab . The principal way for Mayan beekeepers to access nest interiors and extract honey and wax was via the ends of the rustic log hives. Here I argue that the day signs Imix , Kib’ , Kab’an , Kawak and Ajaw represent cross sections of log hives that are visible when opened at their sides. The signs' interiors reflect extant but stylized bee nest elements that are important to beekeeping, such as food stocks, brood content, adult bees, and nest entrances. Similar to all other day signs and nearly all variants, their roundish, outer frames imitate a hollow log’s solid wall. In those cases where numerous hives were densely stacked together in bee sheds, Maya beekeeping must have become more complex in its organization. To tackle increasing complexity in bee management and sustain colony growth and optimize honey and wax production, Mayan beekeepers likely administered their work based on written figures related to beekeeping details. The five day signs were probably derived from a mnemonic system that originally was intended to identify individual log hives, to keep records of colonies and to share information with others involved in this activity.

A genetic and morphological survey to trace the origin of Melipona beecheii (Apidae: Meliponini) from Cuba

The stingless bee Melipona beecheii is extensively distributed across Mexico and Central America and the only Melipona found in the Greater Antilles. The aim of this work was to establish possible continental affinities of M. beecheii from the Antilles. We compared populations from the main island (Cuba) with two clearly separated genetic lineages of M. beecheii on mainland, namely the Yucatan peninsula (Mexico) and Costa Rica. We used morphometrics, the degree of cephalic maculation, and microsatellite variation. Results indicate a greater morphological and genetic similarity between populations from Cuba and the Yucatan peninsula with respect to those of Costa Rica. Based on our findings, we conclude that the origin of M. beecheii from Cuba is more likely the Yucatan peninsula (Mexico). We found evidence that isolation has not been large enough to result in a new genetic lineage.

Antifungal activity of honey from stingless bee Melipona beecheii against Candida albicans

At present there is a growing interest in the search of novel antifungals in natural products. Honey of Melipona beecheii, a stingless bee species, is used to treat a wide spectrum of diseases for centuries. Recently, M. beecheii honey is of great interest in Mexico and Central America due to its biological activities, especially antimicrobial effects. However, antifungal properties of Melipona honey remain poorly explored. The main purpose of this study was to establish the effects of M. beecheii honey treatment against Candida albicans, determining inhibition of growth and changes on cell wall. Agar diffusion assays and broth macrodilution method were employed to determine the antifungal effect of Melipona honey. Also, scanning electron microscopy was performed to study the effects on cell wall of C. albicans. As a result, we obtained that concentrations of 20% (v/v) and above inhibit the growth of C. albicans and induced dramatic changes in the structure and integrity of the cell wall. Remarkably, concentrations of 35% (v/v) caused fungicidal effect in vitro against this opportunistic pathogen. Therefore, this natural product may be an attractive source of new antifungal compounds, useful in treatments of recalcitrant infections.

Actividad antioxidante y quelante de metales de las mieles de Melipona beecheii y Frieseomelitta nigra originarias de Tabasco, México

La miel es un producto alimenticio con alto valor nutricional y potencial farmacológico. La mayoría de los estudios de este producto se han centrado en las propiedades de la miel producida por Apis mellifera, que se ha utilizado en medicina alternativa, destacando por sus actividades antioxidantes, antimicrobianas y antiinflamatorias, entre otras. En este trabajo, se identificó el origen floral, la concentración de proteína soluble, los compuestos fenólicos y la actividad antioxidante y quelante de metales de las mieles producidas por Melipona beecheii y Frieseomelitta nigra,originarias de San Marcos, comunidad de Tenosique en Tabasco, México. Los resultados muestran que la miel producida por F. nigra es de origen polifloral derivada principalmente de la especie Piper sp., aff. Brosimum, Asteraceae, Ziziphus sp., Haematoxylum campechianum, mientras que la producida por M. beecheii fue monofloral (Eugenia sp.). La miel de F. nigra presentó mayor concentración de compuestos fenólicos y mayor efectividad para atrapar los radicales superóxido y DPPH, además de un mejor potencial de quelación del cobre. Por su parte, la miel de M. beecheii presentó mayor capacidad de captación de los radicales ABTS y quelación del hierro; mientras que la capacidad de absorción del radical hidroxilo fue similar para ambas mieles. Este trabajo resalta la importancia de contar con análisis palinológicos y bioquímicos sobre las mieles de las abejas nativas sin aguijón por el potencial terapéutico que tienen y de las cuales, en el caso de algunas especies, no se tiene información.

"Population of Pups and Food Reserves in Hives of Melipona Beecheii Rational Bennett as Basic Factors for their Health"

The Meliponini or stingless form a group of about 500 species described worldwide number could be increased due to the existence of cryptic species (high morphological similarity) and the absence of detailed analysis in most genera.

Pentacyclic triterpenes and other constituents in propolis extract from Melipona beecheii collected in Yucatan, México

Thirteen pentacyclic triterpenes, methyl 3-oxours-12-en-23-oate, marsformosanone, taraxerone, β-amyrenone, α-amyrenone, lupenone, 24-methylencycloartan-3-one, moretenol acetate, β-amyrin acetate, germanicol acetate, 24-methylencycloartanyl acetate, β-amyrin, and α-amyrin were identified in a chloroform-methanol propolis extract from Melipona beecheii. Additionally, were identified in this propolis, hexadecanoic acid, methyl ester, octadecanoic acid, methyl ester and 1-triacontanol. The purification of the propolis extract was carried out using different chromatographic techniques, including vacuum liquid chromatography, gravity column chromatography and gel filtration chromatography Sephadex LH-20. The identification of the metabolites was performed using mass spectrometry.

In vitro antioxidant and anti-inflammatory activities of Melipona beecheii honey protein fractions

Proteins extracted from Melipona beecheii honey were assessed for its antioxidant and anti-inflammatory activity by in vitro assays. Protein content was 3.56 mg/mL and after reverse phase high-performance liquid chromatography fractionation eight distinctive peaks were detected with molecular weight ranged from 7.04 to 95.35 kDa. Low molecular weight fraction constituted by proteins with weights of 7.04, 14.07 and 24.16 kDa, exhibited the highest antioxidant activities with the lowest IC50 values for radical scavenging activity (0.4262 mg/mL), chelating effects (0.3763 mg/mL), and reducing power (0.3060 mg/mL). Same fraction exhibited the lowest IC50 values for inhibition of protein thermal denaturation (0.4115 mg/mL) and cell membrane stabilization (0.5563 mg/mL). For its part, the complete protein extract exhibited the lowest inhibition of proteolytic activity (4.3548 mg/mL). From the results, it is concluded that proteins of honey produced by Melipona beecheii are partially related with its antioxidant and anti-inflammatory activities. These findings validate the use that many Mesoamerican cultures have made of the honey produced by M. beecheii for the healing of lesions and dermic pathologies. Once deeper studies and the pertinent clinical trials have been carried out, the honey of M. beecheii and its proteins could be incorporated into products for pharmaceutical use.

Importancia de las abejas sin aguijón, en las familias del área rural de Guatemala

Las abejas sin aguijón, son originarias de las Zonas Tropicales y Subtropicales del planeta. En el continente Americano están distribuidas desde México hasta Argentina. En América Los Mayas produjeron tecnología, para el manejo de estas abejas, y por la abundancia de vegetación, la abeja real (Melipona beecheii), que produce la miel blanca, la trabajaron en gran escala, cada meliponario constaba de cientos de colonias de abejas. Durante la colonia se exportó la miel de abejas meliponas a España. En Guatemala se trabaja en forma rústicas las abejas de los géneros meliponas y trigonas, existen muy pocos meliponicultores tecnificados. Los Guatemaltecos en el área rural, utilizan las mieles de las diversas especies de abejas nativas, como medicina, aunque la producción de miel es poca los precios de la miel de estas abejas son más altos que la de Apis mellífera. La miel que producen los Meliponicultores de Guatemala, no satisface la demanda que existe, porque existen pocos meliponarios y la mayoría desconocen de las técnicas modernas para la crianza de estas abejas. Estas abejas se trabajan en pequeñas áreas y en el patio de la casa de la familia en el área rural, como son inofensivas, los hijos y la madre, pueden producir miel y polen. Para la nutrición de la familia y vender los excedentes, la miel es rica en proteína, por la presencia del polen, vitaminas, minerales enzimas y energía. En Guatemala los meliponicultores tecnificados utilizan dos tipos de colmena, la Noguera Neto y Portugal Araujo, la primera es utilizada para las abejas pequeñas , mientras que la Portugal Araujo es usada para abeja grande ambas facilitan el manejo de la abeja, durante el presente año se validará una colmena que propongo para las abejas del género Meliponas. Las abejas nativas pecorean flores pequeñas donde no pueden extraer néctar las abejas Apis mellífera, son selectivas y no extraen néctar en plantaciones donde han aplicado herbicidas o insecticidas, la miel es líquida. Las abejas Sin aguijón son resistentes a las enfermedades de las abejas Apis mellífera,los enemigos de las abejas nativas son las hormigas, lagartijas, los sapos y los pájaros y una mosca pequeña de color negro que se introduce a las colonias para ovopositar, y las larvas se alimentas de polen y miel, en Guatemala esta mosca se le llama Nemen.

Estudio cualitativo de las abejas nativas sin aguijón en Jalapa, Guatemala

La meliponicultura, o crianza de las abejas nativas, una actividad asociada a la mayoría de los pueblos indígenas en Mesoamérica. Aunque en los últimos años se han realizado estudios sobre los meliponinos en Guatemala, ninguno de estos se han realizado en Jalapa. El objetivo de este estudio fue documentar la diversidad de abejas utilizadas por los agricultores en Jalapa, los usos de los productos de la colmena y el grado de tecnificación de la crianza. Esta es información básica necesaria para promover la producción de meliponinos en la región de manera racional. Esto se realizó por medio de colecta de las especies, ubicación de las colmenas y realización de encuestas a los agricultores,en los 7 municipios del departamento de Jalapa. Se registraron un total de 163 nidos de abejas sin aguijón en 31 localidades.Se reportan un total de 9 especies de meliponinos: Melipona beecheii, Trigona (Tetragonisca) angustula, Nanotrigonaperilampoides, Partamona bilineata, Scaptotrigona pectoralis, Plebeia pulchra, Melipona yucatanica, Trigonacorvina y Trigona (Geotrigona) Acapulconis. La meliponicultura en Jalapa, no cuenta con tecnificación y se reduce auna meliponicultura artesanal y llena de misticismo. Las especies con mayor importancia en cuanto a producción son M. beecheii, T. angustula y P. bileneata. En los municipios de Jalapa y Mataquescuintla se encontró una mayor diversidad de abejas y de colmenas. Las abejas nativas están bajo amenaza por la pérdida de su hábitat, por lo que su manejo debe ser sostenible y responsable. La meliponicultura es una actividad amigable con el ambiente, importante para la seguridad alimentaria y se relaciona con la cultura popular, por lo que se debe realizar acciones a nivel local para la protección y utilización responsable de las abejas nativas sin aguijón.

Apis mellifera vs Melipona beecheii Cuban polifloral honeys: A comparison based on their physicochemical parameters, chemical composition and biological properties

Several polifloral honeys from the Apis mellifera and Melipona beecheii bee were analyzed to evaluate their pollen profile, physicochemical parameters, chemical composition and their antioxidant and antimicrobial activity. Flavonoids and other phenolic derivatives, 16 compounds in Apis mellifera honeys and 19 compounds in Melipona beecheii honeys, were identified using the HPLC-DAD-ESI-MS/MS analysis. The physicochemical parameters were within the ranges reported for these honeys, showing some of the distinctive characteristics of M. beecheii honeys compared to A. mellifera honeys, such as their high moisture and acidity. M. beecheii honey showed the highest values of total antioxidant capacity and total phenolic, flavonoid, carotenoids, ascorbic acid, free amino acid and protein contents compared to A. mellifera honey. M. beecheii honey also exhibited a higher antimicrobial activity. Our result shows that M. beecheii honeys is an important source of bioactive compounds with relevant biological properties compared to A. mellifera honey.

Antioxidant and anti-inflammatory activities of phenolic compounds isolated from Melipona beecheii honey

ABSTRACTMethanolic extract was obtained from Melipona beecheii honey and quantitative analysis of components like total amount of phenolic compounds, flavonoids and flavonols were estimated. Phenolic acids and flavonoids were fractionated by reverse phase HPLC and assessed for antioxidant and anti-inflammatory activities by in vitro methods. Total phenol content was estimated as 63.22 mg of Gallic acid equivalents/100 g of honey. Total flavonoids and flavonols were found to be 3.61 and 3.16 mg of Catechin or Naringenin equivalents/100 g of honey, respectively. The flavonoids fraction exhibited the highest antioxidant activity, with IC50 values of 0.33, 0.92 and 0.36 mg/mL for radical scavenging activity, chelating effects and reducing power, respectively. Inhibition of albumin denaturation, membrane stabilization and proteinase inhibition assays indicated that flavonoids extract showed anti-inflammatory activity. Phenolic compounds like phenolic acids and flavonoids present in the M. beecheii honey could be partially responsible for their antioxidant and anti-inflammatory activities.

An insight into the antibiofilm properties of Costa Rican stingless bee honeys.

There is an increasing search for antibiofilm agents that either have specific activity against biofilms or may act in synergy with antimicrobials. Our objective is to examine the the antibiofilm properties of stingless bee honeys. Meliponini honeys from Costa Rica were examined along with Medihoney as a reference. All honeys were submitted to a screening composed of minimum inhibitory concentration, inhibition of biofilm formation and biofilm destruction microplate-based assays against a Staphylococcus aureus biofilm forming strain. Dialysis led to the isolation of an antibiofilm fraction in Tetragonisca angustula honeys. The honey antibiofilm fraction was evaluated for protease activity and for any synergistic effect with antibiotics on a Staphylococcus aureus biofilm. The active fraction was then separated through activity guided isolation techniques involving SDS-PAGEs, anion exchange and size exclusion fast protein liquid chromatographies. The fractions obtained and the isolated antibiofilm constituents were tested for amylase and DNase activity. A total of 57 Meliponini honeys from Costa Rica were studied in this research. The honeys studied belonged to the Tetragonisca angustula (n=36) and Melipona beecheii (n=21) species. Costa Rican Tetragonisca angustula honeys can inhibit the planktonic growth, biofilm formation, and are capable of destroying a Staphylococcus aureus biofilm. The antibiofilm effect was observed in the protein fraction of Tetragonisca angustula honeys. The biofilm destruction proteins allowed ampicillin and vancomycin to recover their antimicrobial activity over a Staphylococcus aureus biofilm. The antibiofilm proteins are of bee origin, and their activity was not due to serine, cysteine or metalloproteases. There were 2 proteins causing the antibiofilm action; these were named the Tetragonisca angustula biofilm destruction factors (TABDFs). TABDF-1 is a monomeric protein of approximately 50kDa that is responsible of the amylase activity of Tetragonisca angustula honeys. TABDF-2 is a protein monomer of approximately 75kDa. Tetragonisca angustula honeys from Costa Rica are a promising candidate for research and development of novel wound dressings focused on the treatment of acute and chronic Staphylococcus aureus biofilm wound infections.

Beekeeping from Antiquity Through the Middle Ages.

Beekeeping had its origins in honey hunting-the opportunistic stealing of honey from wild honey bee nests. True beekeeping began when humans started providing artificial cavities within which the bees could build comb for the queen to lay her eggs and the workers could process honey. By 2450 BCE, the Egyptians had developed sophisticated apiculture, and, within two millennia, beekeeping with horizontal hives had spread throughout the Mediterranean. During Europe's Middle Ages, honey and wax became important commodities for trade, and beekeeping in skep, log, box, and tree hives flourished to meet the demand. Other species of honey bees contributed to the development and spread of beekeeping in Asia beginning around 300 BCE. Meanwhile, beekeeping evolved independently in Mesoamerica with the stingless bee Melipona beecheii, as documented by archaeological finds and written accounts that survived Spanish conquest.

Pollination services of Africanized honey bees and nativeMelipona beecheiito buzz-pollinated annatto (Bixa orellanaL.) in the neotropics

Africanized honey bees (AHBs) are the predominant flower visitors of many plants in the neotropics, although little evidence is available on their efficiency as pollinators on native crops. Annatto (Bixa orellana) is a buzz-pollinated neotropical tree. We compared the pollination service provided by AHBs and native Melipona beecheii (Mb) to annatto in the Yucatan. As a result of the different abilities of both species to sonicate, a prediction of the present study is that AHBs on individual visits would result in less efficient pollinators on this crop. A higher frequency of AHBs on flowers (73.8%) compared with Mb (21.3%) was found. However, AHBs deposited significantly less pollen on the stigma and produced less fruits, with fewer seeds and weight, than Mb. A higher pollination index efficiency was obtained for Mb (0.9) compared with AHBs (0.6). AHBs did not sonicate annatto and gleaned the pollen released after Mb visits, which suggests that they act as commensals of the latter. By acting as commensals, AHBs, despite their high abundance, appear to marginally contribute to the pollination of annatto. Studies conducted under scenarios with a differential abundance of AHBs and efficient sonicating species are necessary to test this hypothesis on annatto and other buzz-pollinated plants in the neotropics.