Comparative psychophysics of Western honey bee (Apis mellifera) and stingless bee (Tetragonula carbonaria) colour purity and intensity perception.

Bees play a vital role as pollinators worldwide and have influenced how flower colour signals have evolved. The Western honey bee, Apis mellifera (Apini), and the Buff-tailed bumble bee, Bombus terrestris (Bombini) are well-studied model species with regard to their sensory physiology and pollination capacity, although currently far less is known about stingless bees (Meliponini) that are common in pantropical regions. We conducted comparative experiments with two highly eusocial bee species, the Western honey bee, A. mellifera, and the Australian stingless bee, Tetragonula carbonaria, to understand their colour preferences considering fine-scaled stimuli specifically designed for testing bee colour vision. We employed stimuli made of pigment powders to allow manipulation of single colour parameters including spectral purity (saturation) or colour intensity (brightness) of a blue colour (hue) for which both species have previously shown innate preferences. Both A. mellifera and T. carbonaria demonstrated a significant preference for spectrally purer colour stimuli, although this preference is more pronounced in honey bees than in stingless bees. When all other colour cues were tightly controlled, honey bees receiving absolute conditioning demonstrated a capacity to learn a high-intensity stimulus significant from chance expectation demonstrating some capacity of plasticity for this dimension of colour perception. However, honey bees failed to learn low-intensity stimuli, and T. carbonaria was insensitive to stimulus intensity as a cue. These comparative findings suggest that there may be some common roots underpinning colour perception in bee pollinators and how they interact with flowers, although species-specific differences do exist.

Floral Species Richness Correlates with Changes in the Nutritional Quality of Larval Diets in a Stingless Bee.

Bees need food of appropriate nutritional quality to maintain their metabolic functions. They largely obtain all required nutrients from floral resources, i.e., pollen and nectar. However, the diversity, composition and nutritional quality of floral resources varies with the surrounding environment and can be strongly altered in human-impacted habitats. We investigated whether differences in plant species richness as found in the surrounding environment correlated with variation in the floral diversity and nutritional quality of larval provisions (i.e., mixtures of pollen, nectar and salivary secretions) composed by the mass-provisioning stingless bee Tetragonula carbonaria (Apidae: Meliponini). We found that the floral diversity of larval provisions increased with increasing plant species richness. The sucrose and fat (total fatty acid) content and the proportion and concentration of the omega-6 fatty acid linoleic acid decreased, whereas the proportion of the omega-3 fatty acid linolenic acid increased with increasing plant species richness. Protein (total amino acid) content and amino acid composition did not change. The protein to fat (P:F) ratio, known to affect bee foraging, increased on average by more than 40% from plantations to forests and gardens, while the omega-6:3 ratio, known to negatively affect cognitive performance, decreased with increasing plant species richness. Our results suggest that plant species richness may support T. carbonaria colonies by providing not only a continuous resource supply (as shown in a previous study), but also floral resources of high nutritional quality.

Social bees are fitter in more biodiverse environments.

Bee population declines are often linked to human impacts, especially habitat and biodiversity loss, but empirical evidence is lacking. To clarify the link between biodiversity loss and bee decline, we examined how floral diversity affects (reproductive) fitness and population growth of a social stingless bee. For the first time, we related available resource diversity and abundance to resource (quality and quantity) intake and colony reproduction, over more than two years. Our results reveal plant diversity as key driver of bee fitness. Social bee colonies were fitter and their populations grew faster in more florally diverse environments due to a continuous supply of food resources. Colonies responded to high plant diversity with increased resource intake and colony food stores. Our findings thus point to biodiversity loss as main reason for the observed bee decline.

Revision of the genus Eueupithecia Prout, 1910 from Argentina (Lepidoptera, Geometridae, Sterrhinae).

A new species is described from Argentina: Eueupithecia vollonoides sp. n. and a differential diagnosis from E. cisplatensis Prout, 1910 is given. The genus Eueupithecia Prout, 1910 (Sterrhinae), so far having been retained to be monotypic, includes two species now.

Resources or landmarks: which factors drive homing success in Tetragonula carbonaria foraging in natural and disturbed landscapes?

To date, no study has investigated how landscape structural (visual) alterations affect navigation and thus homing success in stingless bees. We addressed this question in the Australian stingless bee Tetragonula carbonaria by performing marking, release and re-capture experiments in landscapes differing in habitat homogeneity (i.e., the proportion of elongated ground features typically considered prominent visual landmarks). We investigated how landscape affected the proportion of bees and nectar foragers returning to their hives as well as the earliest time bees and foragers returned. Undisturbed landscapes with few landmarks (that are conspicuous to the human eye) and large proportions of vegetation cover (natural forests) were classified visually/structurally homogeneous, and disturbed landscapes with many landmarks and fragmented or no extensive vegetation cover (gardens and plantations) visually/structurally heterogeneous. We found that proportions of successfully returning nectar foragers and earliest times first bees and foragers returned did not differ between landscapes. However, most bees returned in the visually/structurally most (forest) and least (garden) homogeneous landscape, suggesting that they use other than elongated ground features for navigation and that return speed is primarily driven by resource availability in a landscape.

Urban gardens promote bee foraging over natural habitats and plantations.

Increasing human land use for agriculture and housing leads to the loss of natural habitat and to widespread declines in wild bees. Bee foraging dynamics and fitness depend on the availability of resources in the surrounding landscape, but how precisely landscape related resource differences affect bee foraging patterns remains unclear. To investigate how landscape and its interaction with season and weather drive foraging and resource intake in social bees, we experimentally compared foraging activity, the allocation of foragers to different resources (pollen, nectar, and resin) and overall resource intake in the Australian stingless bee Tetragonula carbonaria (Apidae, Meliponini). Bee colonies were monitored in different seasons over two years. We compared foraging patterns and resource intake between the bees' natural habitat (forests) and two landscapes differently altered by humans (suburban gardens and agricultural macadamia plantations). We found foraging activity as well as pollen and nectar forager numbers to be highest in suburban gardens, intermediate in forests and low in plantations. Foraging patterns further differed between seasons, but seasonal variations strongly differed between landscapes. Sugar and pollen intake was low in plantations, but contrary with our predictions, it was even higher in gardens than in forests. In contrast, resin intake was similar across landscapes. Consequently, differences in resource availability between natural and altered landscapes strongly affect foraging patterns and thus resource intake in social bees. While agricultural monocultures largely reduce foraging success, suburban gardens can increase resource intake well above rates found in natural habitats of bees, indicating that human activities can both decrease and increase the availability of resources in a landscape and thus reduce or enhance bee fitness.

Determination of interglycosidic linkages in O-glycosyl flavones by high-performance liquid chromatography/photodiode-array detection coupled to electrospray ionization ion trap mass spectrometry. Its application to Tetragonula carbonaria honey from Australia.

RATIONALE: Tetragonula carbonaria pot-honeys are highly valued as a food source and for their biological activities in Australia, and there is a growing interest to know its composition. Phenolic metabolites, which could be related to their beneficial properties, have not been studied in depth yet. METHODS: Mass spectrometry (MS) coupled to liquid chromatography (LC) is an advanced technique for the study of complex flavonoids present in difficult food matrices that hampers their isolation and purification. This allows the tentative characterization of diglycosides/triglycosides establishing the position of the O-glycosylation on the sugar moiety by the study of the MS data in T. carbonaria pot-honeys from Australia. RESULTS: Their spectra obtained by high-performance liquid chromatography/photodiode-array detection/electrospray ionization ion trap mass spectrometry (HPLC/DAD/ESI-MS(n) ) revealed for the first time 19 quercetin, kaempferol and isorhamnetin O-glycosides. These compounds were clustered in flavonoid triglycosides, diglycosides and monoglycosides. The first cluster contained one flavonoid trihexoside, two -3-O-(2-hexosyl, 6-rhamnosyl)hexosides and their isomers and two -3-O-(2,6-di-rhamnosyl)hexosides. In the second cluster, eleven flavonoid diglycosides such as three -3-O-(2-hexosyl)hexosides, four -3-O-(2-rhamnosyl)hexosides and one -3-O-(6-rhamnosyl)hexoside as well as two -3-O-(2-pentosyl)hexosides and one tentative -3-O-(3-pentosyl)hexoside were detected. In the monoglycoside group, only one flavonoid -3-O-hexoside was identified. CONCLUSIONS: The occurrence of this large number of flavonoid glycosides could be due to the low glucosidase activity previously reported in stingless bee honey.

Bees at war: interspecific battles and nest usurpation in stingless bees.

We provide the first evidence for interspecific warfare in bees, a spectacular natural phenomenon that involves a series of aerial battles and leads to thousands of fatalities from both attacking and defending colonies. Molecular analysis of fights at a hive of the Australian stingless bee Tetragonula carbonaria revealed that the attack was launched by a related species, Tetragonula hockingsi, which has only recently extended its habitat into southeastern Queensland. Following a succession of attacks by the same T. hockingsi colony over a 4-month period, the defending T. carbonaria colony was defeated and the hive usurped, with the invading colony installing a new queen. We complemented our direct observations with a 5-year study of more than 260 Tetragonula hives and found interspecific hive changes, which were likely to be usurpation events, occurring in 46 hives over this period. We discuss how fighting swarms and hive usurpation fit with theoretical predictions on the evolution of fatal fighting and highlight the many unexplained features of these battles that warrant further study.

In vitro antibacterial phenolic extracts from "sugarbag" pot-honeys of Australian stingless bees (Tetragonula carbonaria).

Australian stingless bee honeys have been shown to exert antioxidant and in vitro antimicrobial properties; however their bioactive factors remained unidentified. This study investigated the antibacterial properties of phenolic extracts from Tetragonula carbonaria honeys. Honeys were harvested from beehives in three sites of South East Australia. Liquid-liquid extractions yielded the phenolic concentrates, for analyses by liquid and gas chromatography mass spectrometry. Antibacterial assays were conducted against Staphylococcus aureus and Klebsiella pneumoniae by in vitro agar diffusion and broth dilution assays. The phenolic extracts averaged to 5.87 mg/100 g of raw honeys, and constituents were 3-phenyllactic acid, lumichrome, diglycosylflavonoids, norisoprenoids. The honeys did not contain methylglyoxal, dihydroxyacetone or phenolics characteristic of Leptospermum nectars. Hydrogen peroxide content amounted up to 155.8 µM in honeys. Beside the bactericidal effects of hydrogen peroxide at 760 µM, other antibacterial factors were the phenolic extracts of "sugarbag" honeys that were active at minimum bactericidal concentrations of 1.2-1.8 mg/mL.

Tortricid moths reared from the invasive weed Mexican palo verde, Parkinsonia aculeata, with comments on their host specificity, biology, geographic distribution, and systematics.

As part of efforts to identify native herbivores of Mexican palo verde, Parkinsonia aculeata L. (Leguminosae: Caesalpinioideae), as potential biological control agents against this invasive weed in Australia, ten species of Tortricidae (Lepidoptera) were reared from Guatemala, Mexico, Nicaragua, and Venezuela: Amorbia concavana (Zeller), Platynota rostrana (Walker), Platynota helianthes (Meyrick), Platynota stultana Walsingham (all Tortricinae: Sparganothini), Rudenia leguminana (Busck), Cochylis sp. (both Tortricinae: Cochylini), Ofatulena duodecemstriata (Walsingham), O. luminosa Heinrich, Ofatulena sp. (all Olethreutinae: Grapholitini), and Crocidosema lantana Busck (Olethreutinae: Eucosmini). Significant geographic range extensions are provided for O. duodecemstriata and R. leguminana. These are the first documented records of P. aculeata as a host plant for all but O. luminosa. The four species of Sparganothini are polyphagous; in contrast, the two Cochylini and three Grapholitini likely are specialists on Leguminosae. Ofatulena luminosa is possibly host specific on P. aculeata. Host trials with Rudenia leguminana also provide some evidence of specificity, in contrast to historical rearing records. To examine the possibility that R. leguminana is a complex of species, two data sets of molecular markers were examined: (1) a combined data set of two mitochondrial markers (a 781-basepair region of cytochrome c oxidase I (COI) and a 685-basepair region of cytochrome c oxidase II) and one nuclear marker (a 531-basepair region of the 28S domain 2); and (2) the 650-basepair "barcode" region of COI. Analyses of both data sets strongly suggest that individuals examined in this study belong to more than one species.

Ambient temperature influences Australian native stingless bee (Trigona carbonaria) preference for warm nectar.

The interaction between flowers and insect pollinators is an important aspect of the reproductive mechanisms of many plant species. Several laboratory and field studies indicate that raising flower temperature above ambient can be an advantage in attracting pollinators. Here we demonstrate that this preference for warmer flowers is, in fact, context-dependent. Using an Australian native bee as a model, we demonstrate for the first time a significant shift in behaviour when the ambient temperature reaches 34 degrees C, at which point bees prefer ambient temperature nectar over warmer nectar. We then use thermal imaging techniques to show warmer nectar maintains the flight temperature of bees during the period of rest on flowers at lower ambient temperatures but the behavioural switch is associated with the body temperature rising above that maintained during flight. These findings suggest that flower-pollinator interactions are dependent upon ambient temperature and may therefore alter in different thermal environments.

Composition and antioxidant activity of Trigona carbonaria honey from Australia.

Stingless bees (Tribe Meliponini) are a diverse group of highly eusocial bees distributed throughout the tropics and subtropics. Trigona carbonaria honey, from Australia, was characterized by traditional physicochemical parameters (acidity, sugars, diastase, electrical conductivity, hydroxymethylfurfural, invertase, nitrogen, and water content) and other compositional factors (flavonoids, polyphenols, organic acids, and water activity), as well as total antioxidant capacity and radical scavenging activity. For the Australian T. carbonaria, the traditional analytical parameters were similar to those previously reported for neotropical stingless bee honey and confirm that honeys produced by Meliponini bees possess several physicochemical properties that are distinctly different from Apis mellifera honey, with higher values of moisture (26.5 +/- 0.8 g of water/100 g of honey), water activity (0.74 +/- 0.01), electrical conductivity (1.64 +/- 0.12 mS/cm), and free acidity (124.2 +/- 22.9 mEq/kg of honey) and a very low diastase activity (0.4 +/- 0.5 diastase number) and invertase activity (5.7 +/- 1.5 invertase number). The sugar spectrum was quite different from that of A. mellifera honey, with 20.3 +/- 2.9 g of maltose/100 g of honey. The values of pH (4.0 +/- 0.1), lactonic acidity (4.7 +/- 0.8 mEq/kg of honey), sucrose (1.8 +/- 0.4 g/100 g of honey), and fructose/glucose ratio (1.42 +/- 0.13) fell in the same ranges as those of A. mellifera honey. Citric (0.23 +/- 0.09) and malic (0.12 +/- 0.03) acid concentrations (in g/kg of honey) of T. carbonaria honeys were in the range described for A. mellifera honey. D-Gluconic was more concentrated (9.9 +/- 1.3 g/kg of honey), in the range of Italian Castanea, Thymus, Arbutus, and honeydew honeys. Flavonoid content was 10.02 +/- 1.59 mg of quercetin equivalents/100 g of honey, and polyphenol contents were 55.74 +/- 6.11 mg of gallic acid equivalents/100 g of honey. The antioxidant activity, expressed as percentage of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) cation (ABTS(*+)) decolorization, was 233.96 +/- 50.95 microM Trolox equivalents, and free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH(*)) depletion was 48.03 +/- 12.58 equivalents of ascorbic acid. All reported values are averages +/- standard deviation. The antioxidant activity can represent an important added value for T. carbonaria honey, to initiate a medicinal approach for both nutritional and pharmaceutical applications, besides further physicochemical characterization.

Composition of stingles bee honey: setting quality standards

Se compilaron datos de composición de 152 mieles de abejas sin aguijón (Meliponini) en estudios realizados desde 1964, y se evaluaron para proponer requisitos de calidad para este producto. Dado que la miel de abejas sin aguijón tienen una composición distinta a la de Apis mellifera, algunos parámetros físico-químicos fueron presentados según la especie abejas sin aguijón. El origen entomológico de la miel se asignó a 17 especies de Meliponini de Brasil, una de Costa Rica, seis de México, 27 de Panamá, una de Surinam, dos de Trinidad & Tobago, y siete de Venezuela, mayormente del género Melipona. Los resultados variaron así: humedad (19,9-41,9g/100g), pH (3,15-4,66), acidez libre (5,9-109,0meq/Kg), cenizas (0,01-1,18g/100g), actividad de la diastasa (0,9-23,0DN), conductividad eléctrica (0,49-8,77mS/cm), HMF (0,9-78,4mg/Kg), actividad de la invertasa (19,8-90,1IU), nitrógeno (14,34-144,00mg/100g), azúcares reductores (58,0-75,7g/100g) y sacarosa (1,1-4,8g/100g), El contenido de humedad de las mieles de abejas sin aguijón es generalmente superior al máximo de 20 por ciento establecido para la miel de A. mellifera. Las directrices ofrecidas pueden ayudar a la expansión consistente de la base de datos físico-químicos de miel de abejas sin aguijón, para establecer sus requisitos de calidad en un futuro. El análisis de polen debería dirigirse hacia el reconocimiento de las mieles uniflorales producidas por las abejas sin aguijón, a fin de obtener productos estandarizados según las especies botánicas. Se necesita una campaña de control de calidad de miel tanto para los recolectores de miel de abejas sin aguijón como para los meliponicultores, junto con la armonización de los métodos analíticos