Properties of honey from Tetragonisca angustula fiebrigi and Plebeia wittmanni of Argentina Melina Araceli Sgariglia, Marta Amelia Vattuone, María Marta Sampietro Vattuone, José Rodolfo Soberón, Diego Alejandro Sampietro To cite this version: Melina Araceli Sgariglia, Marta Amelia Vattuone, María Marta Sampietro Vattuone, José Rodolfo Soberón, Diego Alejandro Sampietro. Properties of honey from Tetragonisca angustula fiebrigi and Plebeia wittmanni of Argentina. Apidologie, Springer Verlag, 2010, 41 (6), 10.1051/apido/2010028. hal-00892089 HAL Id: hal-00892089 https://hal.archives-ouvertes.fr/hal-00892089 Submitted on 1 Jan 2010 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Apidologie 41 (2010) 667–675 Available online at: c INRA/DIB-AGIB/EDP Sciences, 2010 www.apidologie.org DOI: 10.1051/apido/2010028 Original article Properties of honey from Tetragonisca angustula fiebrigi and Plebeia wittmanni of Argentina* Melina Araceli Sgariglia1, Marta Amelia Vattuone1,MaríaMartaSampietro Vattuone2, José Rodolfo Soberon´ 1, Diego Alejandro Sampietro1 1 Cátedra de Fitoquímica, Instituto de Estudios Vegetales “Dr. A. R. Sampietro”, Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Argentina 2 Cátedra de Antropología Biológica, Facultad de Ciencias Naturales e Instituto “Miguel Lillo”, Universidad Nacional de Tucumán Ayacucho 471 (4000) San Miguel de Tucumán, Argentina Received 16 September 2009 – Revised 13 February 2010 – Accepted 16 February 2010 Abstract – The composition of honey samples of Plebeia wittmanni (n = 10) and Tetragonisca angustula fiebrigi (n = 12) was analysed. The colours of all collected honeys were amber to dark amber and the pH varied. Moisture was lower than reported for other stingless bee honeys. Conductivity and ash content of P. wittmanni honey were higher than for T. angustula fiebrigi. Acidity of P. wittmanni honey was the highest ever mentioned for all other Plebeia species. Total sugars and sucrose were higher in T. angustula fiebrigi than in P. wittmanni honey. T. angustula fiebrigi honey showed the highest sucrose content ever mentioned and was rich in oligosaccharides. Both honeys split off sucrose, α-glucosides, trehalose, and amylose. The strongest hydrolytic activity was on sucrose, with high activity for T. angustula fiebrigi honey. Raffinose was also hydrolyzed. The honey of both bees inhibited bacterial growth. These properties support, at least in part, the medicinal use of the stingless bee honey by native people. Tetragonisca angustula fiebrigi / Plebeia wittmanni / Meliponini / composition / honey / antibacterial properties 1. INTRODUCTION produce a fine, delicate honey with a delicious flavor (Kent, 1984; van Veen et al., 1990a) More than 400–500 species of Neotropi- that allows their commercialization. The wax cal stingless bees occur in the tropical and of these bees is used for waxing thread, cov- subtropical regions of America (Wille, 1979; ering cheeses, making candles (Kent, 1984). Roubik, 1995; Camargo and Pedro, 2007). But traditionally, these honeys are not consid- Beekeeping with stingless bees, or meliponi- ered as food only. Mesoamerican aborigines culture, was an aboriginal pre-Hispanic prac- use honeys as medicine for treating skin erup- tice in Mesoamerica and their honey was tions (confluent smallpox), tongue sores, urine a well known food resource of the aborig- problems, as emollient for wounds, cracked ines in the North of Argentina (Alderete- lips, skin infections and bruises, and treatment Núñez, 1945). However, the introduction of of upper respiratory system disorders. Diluted Apis mellifera into the region produced a with water it is considered an ophthalmic rem- strong decline in meliponiculture. Different edy (Vit and Tomás-Barberán, 2004). Further- stingless bee species produce honeys with dis- more, it is used as purgative for women who tinct organoleptic properties. Some of them have just given birth and as aid during birth. Corresponding author: M.A. Vattuone, Because of the assigned properties, these hon- [email protected]; [email protected] eys are sold to local pharmacies at a higher * Manuscript editor: Klaus Hartfelder Article published by EDP Sciences 668 M.A. Sgariglia et al. price than A. mellifera honey (van Veen et al., 2.2.2. Acidity and pH 1990b). Several Meliponini species are recorded in Free acidity was determined by titration and pH the North of Argentina, and several are used as was estimated using a digital pH meter Model HI food and medicine. In Tucumán Province, Ar- 8519 (Hanna Instrument). gentina, one may frequently encounter, even in the cities, a small black bee known as “pusquiyo” or “pusquillo” that was identified 2.2.3. Refraction index as Plebeia wittmanni. Another stingless bee from the same region with the common name The measurements were done with an Abbe re- = fractometer (Atago, Japan), all taken at room tem- “angelito” or “rubiecita” ( little blond) was ◦ identified as Tetragonisca angustula fiebrigi. perature (26 C). Before measurements, the refrac- tometer’s sample compartment and window were Despite the frequent occurrence of these sting- first cleaned with acetone. less bees, there are no Argentinean regulations for Meliponini honeys. Some regulations re- Apis mellifera fer to honey (Bianchi, 1990), 2.2.4. Electrical conductivity and these are very similar to the European Di- rections and Regulations, or even more de- The electrical conductivity was measured at tailed, and in accordance with the Codex Al- 25 ◦CusingapH/Conductivity meter Model 20 imentarius, International Honey Commission (Denver Instrument). The instrument was calibrated (CODEX 1969, 1987, 2001). using 0.01 M KCl. In the present study we report on colour, sugar composition, protein content, enzyme activities and antibacterial activity of honeys 2.2.5. Moisture from T. angustula fiebrigi and P. wittmanni of San Miguel de Tucumán, Argentina. For the determination of the moisture content, 2.50 g of each sample was placed in a flat dish and oven-dried at 105 ◦C for three hours, covered, 2. MATERIALS AND METHODS cooled in a desiccator and weighed. The sample was then re-dried for one hour in the oven, cooled and reweighed. The process was repeated at one 2.1. Honey sources hour drying intervals until a constant weight was obtained. Values were expressed as moisture per- P. wittmanni (n = 10) and T. angustula fiebrigi centages. Each analysis was performed in triplicate. (n = 12) honeys were collected around the City of San Miguel de Tucumán, (26◦ 48 30 S65◦ 13 03 W) Argentina. The colonies were transferred from 2.2.6. Ash content logs to wooden boxes of 20 × 20 × 10 cm in the case of P. wittmanni, and 30 × 30 × 30 cm for T. For ash content determination, 2.50 g of each angustula fiebrigi, with removable bottom and lid sample was put in a crucible and dried in an oven parts. Honey sampled during December 2005 and at 105 ◦C for three hours to prevent loss by foam- January 2006 was taken from closed pots by suction ing. After cooling it was ashed overnight in a Muf- with a syringeand immediately processed. fle oven at 600 ◦C. After cooling it was weighed to a constant weight. 2.2. Physicochemical properties 2.3. Sugar determinations 2.2.1. Colour 2.3.1. Mono and disaccharides The colour of the samples was determined using Total neutral sugars were determined by the phe- the Lovibond comparator. nol sulfuric acid method (Dubois et al., 1956) with Studies on Meliponini honey from Argentina 669 glucose as standard. Reducing sugars were deter- buffer (pH 5.2), 40 μL distilled water, and 10 μL mined by the Somogyi-Nelson method (Nelson, of 0.6 M sucrose or 50 μLof0.6Mraffinose, 1944; Somogyi, 1945) using glucose (Sigma- trehalose or 10% amylase, respectively (Sigma- Aldrich, Saint Louis, Missouri, USA) as standard. Aldrich, USA) in a final volume of 0.1 mL. The as- Sucrose and fructose (Sigma-Aldrich, USA) were says were performed at 37 ◦C for 30 min. Reactions determined by the resorcinol method of Cardini were stopped by heating at 100 ◦Cfor2min. et al. (1955) using sucrose as standard. Glucose The enzymatic hydrolysis and quantification was determined by the glucose-oxidase-peroxidase of sugars was determined as reducing power re- (Sigma-Aldrich, USA) coupled assay (Jorgensen lease when sucrose or raffinose is used as sub- and Andersen, 1973) with glucose as standard. strate (Nelson, 1944; Somogyi, 1945); by the re- sorcinol method (Cardini et al., 1955), and/or by the specific measurement of glucose and fructose 2.3.2. Oligosaccharides produced by the glucose-oxidase-peroxidase assay (Sigma-Aldrich, USA) (Jorgensen and Andersen, A diluted honey sample (0.5 g of honey in 1973), and by the enzymatic assay of D-fructose 2 mL of distilled water) was adsorbed onto a 10 × dehydrogenase (Sigma-Aldrich, USA), respectively 2.2 cm carbon-Celite column. The column was (Ameyama, 1982). α-glucosidase activity was de- washed with 2 L of distilled water to eliminate solu- tected by reducing power release after enzyme ex- ble sugars and finally oligosaccharides were eluted tract was incubated with amylose as substrate. with 70% ethanol (Merck, Damstadt, Germany). Fractions of 1.6 mL were collected and pooled. Oligosaccharides were determined as total sugars 2.6. Antibacterial activity (Dubois et al., 1956). 2.6.1. Microorganisms 2.4. Proteins The microorganisms used included bacterial strains isolated from skin wounds at Hospital “Nicolás Avellaneda”, SIPROSA, Tucumán, Ar- Proteins were determined by the method of gentina. Gram negative bacteria: Escherichia coli Lowry et al. (1951) using bovine serum albumin (IEV301), Pseudomonas aeruginosa (IEV 305).