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

OBJECTIVE: 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. METHOD: 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. RESULTS: 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. CONCLUSION: 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.

Evaluación de la posible adulteración de mieles de abeja comerciales de origen costarricense al compararlas con mieles artesanales provenientes de apiarios específicos / Evaluation of the potential adulteration of commercial honey distributed in Costa Rica compared with artesanian honey samples coming from specific apiaries

Honey is the principal apiculture product, produced by Apis mellifera bee. This, as any other food product, has to accomplish certain quality standards, including physicochemical, organoleptic and microbiological properties. Within these parameters, different measures are considered as adulteration indicators, including hydroxymethylfurfural and diastase enzyme activity which are associated to overheating, and the sucrose, glucose and fructose content. In this study, a determination of previous parameters, additional to glucose, fructose and fructose/glucose index was performed to 35 artisan samples, obtained directly from beekeeper, previously characterized as having good productive practices and 25 commercial samples. Same time, the results obtained were compared and interpreted, in order to determine the kind of adulteration present in the honey sample. The 89% of artesian samples accomplished the parameters established by the Codex Alimentarius for HMF content, diastase activity, sucrose and simple sugars content. Nevertheless, only 20% (5) of the commercial samples accomplished the international and national normative. 24% of these samples presented succrose addition, 32% overheating or inverted sugar syrup addition, and 24% were adulterated with inverted sugar syrup.

La miel es el principal producto de la apicultura, generado por la abeja Apis mellifera. Éste, como cualquier producto alimenticio, debe cumplir con ciertas normas de calidad, propiedades fisicoquímicas, organolépticas y microbiológicas. Entre estos parámetros se encuentran el contenido de hidroximetilfurfural (HMF), indicador de adulteración, sobrecalentamiento y envejecimiento de la miel, la actividad de la enzima diastasa, indicador de sobrecalentamiento, y la determinación de contenido de sacarosa y azúcares simples, indicadores de adulteración. En el presente estudio, se procedió a determinar los anteriores parámetros, además del contenido de glucosa, fructosa e índice fructosa/glucosa a 35 muestras de mieles artesanales obtenidas directamente del apicultor, previamente catalogado como productor con buenas prácticas y 25 de mieles distribuidas en expendios. A su vez, se compararon los resultados poblacionales obtenidos y se les interpretó, con el objetivo de determinar el tipo de alteración que presentara la miel. Se determinó que 89% de las mieles control cumplieron con los parámetros establecidos por el Codex Alimentarius para su contenido de HMF, actividad de diastasa, contenido de sacarosa y de fructosa y glucosa. Contrario, sólo el 20% (5) de las mieles comerciales logró cumplir los estándares internacionales y la normativa nacional. Se destaca que un 24% de las muestra comerciales presentan adición de sacarosa y glucosa, 32% sobrecalentamiento o adulteración con sirope de azúcar invertido, y 24% adición de jarabe de azúcar invertido.