Bioactive properties of honeys from stingless bees and Apis mellifera bees in the food industry

Abstract The "medicinal" benefits of honey have been known for thousands of years, being used as an antiseptic, fortifier, soother, healer, laxative, among others. But in addition to its use as a therapeutic, there is evidence that since ancient Rome honey has been used as a food preservative. Currently, the food industry faces several challenges, among which are maintaining the quality and safety characteristics of their products with the minimum addition of artificial ingredients and additives due to the worldwide increase in demand for natural products and functional properties, as well as helping to reduce food loss and thus its ecological footprint. Currently, honey produced by stingless bees has been the subject of research, due to its nutritional value and its bioactive properties of great utility, both in the pharmaceutical industry and in food production. Therefore, a review of published articles describing the potential uses of the bioactive properties of honey, mainly that produced by stingless bees, in the food industry was carried out.


Introduction
Recently, consumers have begun to pay more attention to the components of their food, where they come from, and how they are made. Within the food sector, there is a growing attitude toward going back to basics. Both consumers and manufacturers are moving towards "less processed" foods made from easily understood ingredients without flavours, colours or synthetic additives (Nascimento et al., 2018), especially when these food additives are thought to be harmful to health, due to the large amount of information received through the media about the improper use practices of the additives that may be detrimental to the health of consumers (Dur an, 2001). Despite being safe and legal ingredients that have been used in food for decades without any objective evidence of adverse health effects (Cassiday, 2017). However, minimally processed foods, in other words, without chemical preservatives or other artificial additives, should remain of high quality and have a long shelf life (Nascimento et al., 2018).
Diversely, cases of obesity and diabetes are increasing. Chronic diseases cause approximately 41 million deaths worldwide each year (FAO-WHO, 2018). Consumers are looking for product innovation, preferring those that provide a health benefit. The market for ingredients is prospering, with the so-called functional ingredients gaining special prominence, as they provide nutritional value combined with a beneficial effect on health, and mainly with convenience. The most important challenge is to ensure that functional ingredients survive and remain "active" and "bioavailable" even after processing and storage (Vicentini et al., 2016). This is why there is currently a continued interest in incorporating honey into foods because of its "healthy" and natural image (Bakr et al., 2015).
Honey is a natural substance produced by bees from the nectar of flowers or secretions of living parts of plants or excretions of plant sucking insects which bees collect, transform and combine with specific substances of their own, and deposit, dehydrate, store and leave in the comb to mature or age (Codex Alimentarius, 2001).
In addition to the most popular use of honey as a natural sweetener, it is also used in traditional medicine because of its antimicrobial, antioxidant, antiviral, antitumor, and anti-inflammatory properties (Khan et al., 2018;Ruiz-Navajas et al., 2011;Viuda-Martos et al., 2008).
Apis mellifera honey may have more than 180 compounds (Gheldof et al., 2002), but it is essentially a supersaturated solution of different sugars, including fructose (38.3%), glucose (30.3%), maltose (7.1%), and sucrose (1.3%). Other substances found in honey include acids (0.5%), proteins (0.3%), minerals (0.2%), and trace components. The moisture content of honey is about 17.7 g/100 g, its total acidity 0.08 g/ 100 g and its ash content is about 0.18 g/100 g (Nagai et al., 2006). The wide variety of minor components includes phenolic acids and flavonoids, ascorbic acid, enzymes (catalase and peroxidase) carotenoids and products of the Maillard reaction, all of which are considered to be the main participants in the antioxidant potential of honey (Abu Bakar et al., 2017;Sarmento Silva et al., 2013).
Since stingless bee honey has a different composition than A. mellifera honey, and due to the great heterogeneity of stingless bee species, among other factors, their composition can change frequently, being the moisture the main difference in their composition, presenting a percentage higher than 20% with respect to that accepted for A. mellifera honey (Souza et al., 2006).
With respect to the antimicrobial potential of A. mellifera honey, it has been determined that several elements are involved (Cooper et al., 2002). The antimicrobial activity of many types of honeys has been predominantly attributed to their hydrogen peroxide concentration (Irish et al., 2008), as this activity is observed to be decreased after the addition of the enzyme catalase to these honeys. However, other types of honeys, such as Manuka honey and honey produced by stingless bees, remain active after treatment with catalase, indicating that other substances than hydrogen peroxide contribute greatly to the antimicrobial activity of honey (Boorn et al., 2010;Cooper et al., 2002).
Currently, the food industry uses A. mellifera honey in its processes as an ingredient because it favors some of the characteristics required during food manufacturing such as moisturization, viscosity, flavor, color, hygroscopicity, miscibility and spreadability.
In meat products such as hams, bacon and sausages, A. mellifera honey enhances the flavours of the meat and spices when mixed with brine, as it allows the ingredients to be bound together and is a growing medium in cured products. In meat cuts intended for roasting it reduces the formation of heterocyclic aromatic amines and their mutagenic effects. Similarly, A. mellifera honey can improve cooking performance in poultry meat, since because meat products are sold by weight, yields are very important for the meat industry (Otles, 2006).
However, A. mellifera honey is not only used in the meat industry, but also as a sweetener in beverages, moisture retainer in bakery and pastry, stabilizer in ice cream, spice and herb intensity enhancer in marinades, and acidity neutralizer in sauces and dressings, among others (Bogdanov, 2016).

Honey as an antioxidant
Honey contains both aqueous and lipophilic antioxidants, substances that have as their main mechanism of action the donation of an electron to free radicals, neutralizing, decreasing or eliminating their ability to damage cells and the main biomolecules, such as nucleic acids, proteins and lipids (Lobo et al., 2010); the interaction between them makes honey an ideal product as a natural antioxidant that would allow the replacement of synthetic compounds in food systems to improve consumer perception (Ruiz-Navajas et al., 2011).
Oxidative deterioration is one of the main culprits in the reduction of quality and acceptability of food products. This process can be initiated by the action of high temperatures, ionizing radiation, light, metal ions, chlorophyll or other pigments, exposure to the enzyme lipoxygenase or to metalloprotein catalysts. The effect of lipid hydroperoxides and their breakdown products has been reported as harmful on food since it implies a significant decrease in the nutritional value of food when the loss of vitamins and essential fatty acids is observed. Oxidation also affects sensory quality by changing colour, texture and taste, which shortens shelf life and may result in consumer rejection. In addition, the accumulation of these substances is related to damage to biological tissues and the presence of degenerative diseases.
In order to control the oxidation processes of foods, technologists have adopted a series of strategies that involve an appropriate choice of raw materials, processing, packaging material and storage conditions (Delgado, 2004) and it is often necessary to use antioxidants, such as nucleic acids, proteins and lipids (Lobo et al., 2010), so when they are added to the product, they contribute to maintaining the organoleptic characteristics and preserving the nutritional quality of the food, as well as helping to considerably preserve the health of the individuals who consume them (Guerrero, 2015).
The use of antioxidants has been widely discussed because they are quite volatiles, and decompose easily at high temperatures and because of their potential toxic effect on health (Valenzuela & P erez, 2016). Consequently, there has been a growing interest in the search for natural antioxidants to replace synthetic substances, honey being one of them, as well as royal jelly and propolis, mainly due to their content of phenolic compounds (Viuda-Martos et al., 2008).
Phenolic compounds or polyphenols are secondary metabolites produced by plants, whose concentration can be observed in different structures: fruits, flowers, stems, leaves, roots, seeds and barks (Valencia-Avil es et al., 2017), in which their biosynthesis is regulated by different enzymes, depending on the species of the plant, its needs and the oxidative stress to which it is subjected (da Silva et al., 2016;Dezmirean et al., 2017).
There are about 8000 identified phenolic compounds and most of these have a 3-ring structure, two aromatics and one oxygenated heterocycle (Mercado-Mercado et al., 2013). In general, phenolic compounds have been classified into two major groups: flavonoids and non-flavonoids (Valencia-Avil es et al., 2017).
Flavonoids are classified according to the degree of unsaturation and oxidation of the C-ring. Flavonoids in which the B ring is linked in position 3 of the C ring are called isoflavones. Those in which the B ring is linked in position 4 are called neoflavonoids, while those in which the B ring is linked in position 2 can be further subdivided into flavones, flavonols, flavanones, flavanonols, flavanols or catechins, anthocyanins and chalcones (Panche et al., 2016). The most abundant flavonoids in honey are: flavones, flavonols and flavanones, such as apigenin, chrysin, galangin, genistein, kaempferol, luteolin, pinobanksin, pinocembrine and quercetin (Table 1) (Cianciosi et al., 2018). While in the non-flavonoid group there are hydroxybenzoic acids, hydroxycinnamic acids, volatile polyphenols, stilbenes and various compounds (lignans and coumarins) (Valencia-Avil es et al., 2017), among the most commonly identified in honey are caffeic and benzoic acids (Moniruzzaman et al., 2014).
The antioxidant activity of flavonoids results from a combination of iron chelating and free radical scavenging properties, together with the inhibition of enzymes responsible for producing superoxide anions (lipoxygenase, cyclooxygenase, myeloperoxidase, nicotinamide adenine dinucleotide phosphate oxidase and xanthine oxidase), thus avoiding the generation in vivo of reactive oxygen species, as well as organic hydroperoxides. Similarly, it has been established that another way in which flavonoids interfere with the formation and propagation reactions of free radicals is through the inhibition of enzymes not directly involved in oxidative processes, such as phospholipase A2, as well as the stimulation of others with recognized antioxidant properties such as catalase and superoxide dismutase (Clappini et al., 2013;).
The Folin-Ciocalteu test is one of the most widely used to determine the concentration of total phenolic compounds in foods, which is based on the fact that phenolic compounds reduce the phosphomolibdotungstic acid present in the Folin-Ciocalteu reagent, at basic pH, giving rise to a blue coloration that can be quantified spectrophotometrically at 765 nm based on a standard straight line of gallic acid (Garc ıa et al., 2015).
To determine the antioxidant activity there are several methods, in vitro or in vivo, but one of the most applied to know the total antioxidant capacity of a compound, mixture or food in vitro, consists of quantifying this antioxidant activity in relation to the loss of color of chromogenic substances of a radical nature, where ABTS (2,2 0 -Azino-bis(3-ethylbenzothiazolin-6-sulphonic acid), DPPH (2,2-diphenyl-1-picrylhydrazyl), FRAP (ferric reduction antioxidant) and ORAC (oxygen radical absorbance capacity) are the most used (Ahmed et al., 2018;Kuskoski et al., 2005;Moniruzzaman et al., 2012).
It has been reported that honey produced by stingless bees has a high content of total phenolic compounds and therefore a higher antioxidant capacity compared to other types of honeys (Rodr ıguez et al., 2007), which is confirmed by Alvarez-Suarez et al. (2018) when observing that Melipona beecheii honey showed higher values regarding the antioxidant capacity, total phenolic compounds, flavonoids, carotenoids, ascorbic acid, free amino acids and proteins in comparison with A. mellifera honey, as shown in Table 2.
Da  identified taxifoline (C 15 H 12 O 7 ) in samples of honey produced by Melipona (Michmelia) seminigra merrillae, collected in the central and southern region of Amazon's state in Brazil, a flavonoid that had not been previously described in honeys from stingless bees. Biluca et al. (2017) reported values between 10.3 and 98.0 m/100 g gallic acid equivalent (GAE) of total phenolic compounds and 1.41 and 18.5 mg/100 g ascorbic acid equivalent (EAA) as antioxidant activity (DPPH and FRAP), in honey samples from 10 stingless bee species, including the genera Melipona, Scaptotrigona, Tetragonisca, Trigona and Tetragona. While out of 26 phenolic compounds detected, relevant amounts of some phenolic and flavonoid acids (mandelic acid, caffeic acid, chlorogenic acid, rosmarinic acid, aromadendrin, isoquercitrin, eryiodicthiol, vanillin, umbeliferone, syringaldehyde, synapaldehyde, carnosol) were described for the first time, showing a significant correlation with the antioxidant capacity of honey. Oliveira et al. (2017) contrarily, pointed out to have detected a range between 30.71 mg/100 g and 854.02 mg/100 g gallic acid equivalent (GAE) in a study where honey samples from six Melipona species were included. Also, in all the honey samples analyzed, high content of total flavonoids was found, ranging from 30.24 mg/100 g quercetin equivalent (QE) to 279.73 mg/100 g. Antioxidant activity measured by the DPPH method showed values ranging from 25.39 mg/mL to 51.44. Of relevance in this study, three important flavonoids were identified for the first time as major compounds in the samples of Table 1. Most common phenolic compounds identified in honey (Cianciosi et al., 2018 Several researchers have observed that the content of total phenolic compounds, flavonoids and carotenoids is higher in darker honeys (Montenegro & Mej ıas, 2013) and therefore, their antioxidant activity is also higher (Alvarez-Suarez et al., 2010;Bertoncelj et al., 2007;Ku s et al., 2014b;Vela et al., 2007). Similarly, single-flower honeys have a higher phenolic content than multi-flower honeys (Ruiz-Ruiz et al., 2017). S anchez-Chino et al. (2019) described that polyfloral honey (Piper sp., aff. Brosimum, Asteraceae, Ziziphus sp. and Haematoxylum campechianum) from Frieseomelitta nigra presented higher concentration of phenolic compounds and soluble proteins than unifloral honey (Eugenia sp.) from M. beecheii, which could be due to higher quantity and diversity of pollen; therefore, it could be stated that stingless bees monofloral honeys do not have higher phenolic content, but rather specific phenolic profiles related to the plant of origin (Habib et al., 2014a), as it has been reported by Muñoz et al. (2014) when determining the content of total phenolic compounds, flavonoids and antioxidant activity of 12 A. mellifera honey samples of different floral origin.
The chemical composition and antioxidant properties of honey are directly related to its geo-botanical origin, environmental conditions, post-harvest, processing and storage, which has produced interest in identifying and quantifying these bioactive compounds, not only to determine the quality of the honey (Biluca et al., 2016;Montenegro & Mej ıas, 2013), but also to use them as markers of geographical origin (Alqarni et al., 2014;Alvarez-Suarez et al., 2012), as demonstrated by Montenegro and Mej ıas (2013) with Chilean Quillay A. mellifera honey and Jandaira honey of Brazilian origin.
However, it has been demonstrated the role of phenolic compounds as responsible for the antioxidant properties of honey, it has been proved that, after the extraction of these compounds, there are others like proline (Saxena et al., 2010) that maintain the antioxidant activity of honey, which explains why the extracts have less antioxidant capacity (Ferreira et al., 2009). According to Aljadi and Kamaruddin (2004), the antioxidant capacity of honey is mainly due to the phenolic compounds it contains, although a synergic action between several compounds, such as enzymes, amino acids and  carotenoids also contributes to this capacity (Bouayed & Bohn, 2010). Due to its antioxidant activity A. mellifera honey is used as an additive in a variety of foods and beverages, as it has been shown to act against lipid oxidation of ground turkey meat (Johnston et al., 2005;McKibben & Engeseth, 2002), beef jerky (Nagai et al., 2006); sausages (Mohammed et al., 2013); enzymatic browning of fruits and vegetables (Chen et al., 2000;Jeon & Zhao, 2005) and juices (Gacche et al., 2009), dressings and oil-rich foods (Rasmussen et al., 2008); prevention of heterocyclic aromatic amine formation in beef and chicken subjected to high cooking temperatures (Shin & Ustunol, 2006).
Diversely, Krushna et al. (2007) confirmed the activity of A. mellifera honey as a safe and effective preservative in milk samples, since honey provides hydrogen peroxide as a substrate for the lactoperoxidase enzymatic complex to catalyze the oxidation of the thiocynate ion (SNC-), originating several products with antimicrobial activity, such as hypothiocyanite (OSNC-), thus promoting the preservation of milk. This is confirmed by Dzomba et al. (2013) having fortified raw cow milk with Hypotrigona squamuligera honey extracts and observed that its antioxidant properties were maintained for 5 days, thus maintaining its nutritional integrity.

Honey as an antimicrobial
The antibacterial effect of honey, both bacteriostatic and bactericidal, is mainly against Gram-positive bacteria, many of which pathogenic (Saranraj & Sivasakthi, 2018) are and antimicrobial resistant (Aween et al., 2014;Maddocks & Jenkins, 2013;Murugan, 2012). This has been proven not only in the therapeutic area but also in the food industry. Mundo et al. (2004) concluded that A. mellifera honeys from different floral sources and geographical locations, mainly from the United States (Table 3), inhibited the growth of seven spoilage microorganisms (Alcaligenes faecalis, Aspergillus niger, Bacillus stearothermophilus, Geotrichum candidum, Lactobacillus acidophilus, Penicillium expansum, Pseudomonas fluorescens) and foodborne pathogens (Bacillus cereus, Escherichia coli O157: H7, Listeria monocytogenes, Salmonella enterica Ser. Typhimurium and Staphylococcus aureus).
Several investigations have attributed the antimicrobial activity of honey to its inherent physical-chemical characteristics such as high sugar concentration, high viscosity and osmotic pressure (Libonatti et al., 2014), however, when comparing the antibacterial activity of A. mellifera honey with a sugar-saturated solution in the same proportion, the result showed that the sugar solution exhibited a lower degree of antibacterial activity against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella Typhi, Shigella shiga, Klebsiella aerogenes, Proteus vulgaris and Proteus mirabilis in relation to the observed activity of the honey (Mulu et al., 2005). Besides the fact that the water activity (aw) values of bee honey are between 0.56 and 0.62, which prevents the growth of almost any microorganism except for some yeasts and osmophilic bacteria, the honey has an acid pH, 3.5-4.5, due to the presence of organic acids, which also represents an important antimicrobial factor in food (Estrada et al., 2005;Mothershaw & Jaffer, 2004).
However, there are other components involved in the antibacterial effect of honey, which have been classified into two types of "inhibitors" according to the way they exert their activity: hydrogen peroxide (H 2 O 2 ) and non-peroxide compounds (Taormina et al., 2001).
H 2 O 2 is released like gluconic acid from the breakdown of glucose molecules by the action of the glucose oxidase enzyme secreted by the hypopharyngeal glands of bees, which breaks down glucose molecules. Some researchers have determined that H 2 O 2 is the main antibacterial agent in honey. However, others found that the anti-bacterial activity not related to peroxide is more important because the enzyme responsible for the production of H 2 O 2 is sensitive to light, heat and the presence of oxygen (Bogdanov, 1997), so during the handling and storage of honey this factor could be diminished and therefore the amount of H 2 O 2 released would be insufficient to inhibit bacterial growth (Libonatti et al., 2014). It has also been suggested that the presence of catalase coming from pollen, nectar or released by microorganisms present in honey could contribute to the decrease of H 2 O 2 concentration (Kwakman & Zaat, 2012). Contrarily, the factors not related to H 2 O 2 , such as pH, osmolarity, presence of lysozyme and other substances, as they are not sensitive to light or heat, remain intact even during the storage of honey for long periods (Bogdanov, 1997).
With regard to non-peroxide compounds, it has been proved that substances such as methylglioxal (MGO C 15 H 12 O 7 ), defensin-1 and phenolic compounds are also involved in antimicrobial activity.
Methylglioxal is a compound detected in Manuka honey, native to New Zealand, which is produced by a non-enzymatic conversion of dihydroxyacetone (DHA) present in exceptionally high concentrations in the nectar of manuka tree (Leptospermum scoparium) flowers (Adams et al., 2008) and with activity correlated between its concentration in honey and bacterial inhibition.
Although bees have been shown to produce four types of antimicrobial peptides (himenopecin, defensin-1, apidaecin and abaecin), Kwakman et al. (2010) identified for the first time in honey, the Table 3. Bacterial sensitivity and geo-botanical origin of A. mellifera honey (Mundo et al., 2004 antimicrobial peptide defensin-1 (royalisin), previously identified in royal jelly, as well as in haemolymph, head and hypopharyngeal glands of A. mellifera bees; peptide of which a potent antimicrobial activity is observed, but only against Grampositive bacteria, including B. subtilis, S. aureus and Paenibacillus larvae (Bachanov a et al., 2002). However, it has also been shown that even after neutralizing H 2 O 2 and degrading the protein compounds in honey, it continues to exert antibacterial activity, related to the presence of phenolic compounds (Kwakman et al., 2010). Several researchers have concluded that the antibacterial behavior of honey is mainly associated with such content, since when analyzing the methanolic extract of honey from Ulmo (Eucryphia cordifolia), Quillay (Quillaja saponaria), Avellano (Gevuina avellana) and Tiaca (Caldcluvia paniculata); a better antibacterial capacity was observed than the honeys of origin (Montenegro & Mej ıas, 2013). In addition, an inverse relationship was found between the polyphenol content and the H 2 O 2 concentration (Al-Waili et al., 2013).
Several phenolic compounds have been identified in honeys, however, the activity of these isolated compounds is too low to contribute substantially to the antibacterial effect and it is thought that it is the combination of these compounds that could exert a better effect (Kwakman & Zaat, 2012). Ciulu et al. (2016) presented a review of some research in which the detection of phenolic compounds in different types of honeys has been carried out through the technique of high performance liquid chromatography (Supplementary Table S1).
Although the best characterized honeys are unifloral, multifloral honeys have also been found to have antibacterial activity, highly correlated to their phenolic content, against Staphylococcus aureus, Enterococcus faecalis, Pseudomona aeruginosa, Escherichia coli, Morganella morganii and Klebsiella pneumoniae (Isla et al., 2011). Similarly, as observed in their antioxidant activity, the effectiveness of phenolic compounds against bacteria can be affected by geobotanical origin, climate, unintentional or deliberate heat treatment during processing, and the age of the honey and the method of extraction of the same phenolic compounds during processing (Silici et al., 2010;Stephens et al., 2010).
It should be mentioned that one of the strategies to inhibit bacterial growth in food is to prevent interaction between bacteria and the matrix, thus decreasing food spoilage. An analysis of European monofloral honeys showed an interesting correlation between the antibacterial effect of A. mellifera honey and its ability to inhibit bacterial cell communication (quorum sensing) (Montenegro & Mej ıas, 2013), retarding the expression of, MvfR, and rhl regulons, as well as their associated virulence factors (Ahmed et al., 2018). However, in this case, total phenolic compounds were not directly related to this capacity (Montenegro & Mej ıas, 2013).
As for stingless bee honeys, it has been confirmed that they show a more efficient antimicrobial capacity compared to A. mellifera (Alvarez-Suarez et al., 2018;Chan-Rodr ıguez et al., 2012;Cooper et al., 2002), which may be due to the fact that meliponid honeys tend to be more acidic than apis honey, but mainly to the differences in chemical composition between both (Zamora & Arias, 2011). The latter has been related to the fact that meliponid honey is stored in pots made of cerumen (Abd Jalil et al., 2017), a material with which the bees build and seal their hives, made from the mixture of propolis collected by the bees and wax secreted from specialized glands that the workers have in the dorsal part of their abdomen (Arnold et al., 2018;Nates-Parra, 2001), which makes some researchers think that the antimicrobial effect can be increased, since propolis, being resins from trees (Dard on & Enr ıquez, 2008), contains substances such as phenolic compounds that give it different bioactive properties (Aljadi & Kamaruddin, 2004;Estevinho et al., 2008;Ka c aniov a et al., 2009;Rao et al., 2016), which explains their widespread use in popular medicine (Mercês et al., 2013).
Regarding the action spectrum of stingless bee honeys, some researchers have reported that Grampositive bacteria may be more sensitive compared to Gram-negative bacteria ( Avila et al., 2018;Sgariglia et al., 2010). However, other studies have shown that honeys such as Melipona compressipes manaosensis exhibit different effects on their antibacterial activity with Gram-positive and Gram-negative bacteria, depending on the season in which the honey was collected (wet or dry); presenting a higher activity in honey collected in the dry season (Pimentel et al., 2013). Likewise, it has been shown that the action spectrum varies significantly among species (Nweze et al., 2016).

Honey as a probiotic and prebiotic
Although honey has been considered a microbiologically safe food due to its composition, it is important to note that it is not sterile, as microorganisms such as bacteria, yeasts and, in particular, molds have been found to eventually enter the food chain at an early stage of honey production, for example through pollen (Grabowski & Klein, 2017). However, most of these microorganisms cannot proliferate, remaining dormant due to the antibacterial activity of honey (Olaitan et al., 2007). In some studies, the pathogenic microorganisms that have been isolated have been few, emphasizing the presence of Clostridium botulinum spores, associated with infantile botulism.
For the production of honey, bees ingest nectar and transform it with the help of enzymes, incorporating some symbiotic microorganisms associated with their gastrointestinal system that act as probiotics (Luchese et al., 2017;Salgado et al., 2017), that is, when administered in adequate quantities they exert a beneficial effect on the health of the host, improving the function of the intestinal barrier, digestive processes and stimulating the immune system (Garrote & Bonnet, 2017). Bacteria such as Gluconobacter oxydans, Lactobacillus spp., Pseudomonas spp. and Bacillus spp. have been isolated from Apis honey harvested directly from the hive (Salgado et al., 2017;Tajabadi et al., 2013).
Besides the probiotic effect, microorganisms associated with honey have been studied and have been identified as responsible for producing substances with antimicrobial potential: bacteriocins, substances capable of reducing or eliminating pathogenic competing microorganisms (Salgado et al., 2017). In 2013, a study was carried out with a new strain of bacteria isolated from A. mellifera honey, capable of producing bacteriocine fungicides, showing an inhibitory character against A. niger, Fusarium oxysporum, Pythium and Botrytis cinerea . Lactobacilli such as Lactobacillus helsingborgensis and L. kunkeei showed effect against E. coli and Salmonella enterica (Veress et al., 2016). While Pajor et al. (2018) confirmed that Polish honeys of different floral origins are a potent source of bacteria belonging to the bacillus group, producing metabolites against S.aureus, S. epidermidis, E. coli, L. monocytogenes, P. aeruginosa and C. albicans, which have a potential in the area of food biopreservation.
To increase the development of probiotics, it is necessary to consume substances that promote their development, known as prebiotics, which are indigestible food components that selectively stimulate the growth and/or activity of beneficial intestinal microbiota (probiotics such as lactobacilli and bifidobacteria) for the host (Creus, 2004;Quigley, 2019;Ulrich et al., 2016).
Regarding honey produced by stingless bees, Bogdanov (1997) stated that in Melipona honeys (M. compressipes compressipes, M. favosa favosa and M. trinitatis) maltose was the main disaccharide detected, while Trigona honeys had small but measurable amounts of turanose, trehalose and erlose. Due to its polysaccharides, honey has been the focus of attention in many researches in relation to its potential prebiotic activity. Shamala et al. (2000) showed that the number of Lactobacillus acidophilus and Lactobacillus plantarum increased from 10 to 100 times more in the gut of rats in the presence of A. mellifera honey compared to sucrose. El-Arab et al. (2006) described that A. mellifera monoflower cotton honey increased the amount of endogenous probiotic bacteria (bifidogenic effect) which inhibited the harmful and genotoxic effects caused by mycotoxins, so honey could be a good substitute for sugar in processed foods. Bezerra et al. (2018) concluded that consumption of honey produced by Mimosa quadrivalvis L. and Melipona subnitida D, improved glucose tolerance and reduced total cholesterol and low density lipoprotein (LDL) levels in dyslipidemic rats, as well as increased Bifidobacterium spp. and Lactobacillus spp. counts and preserved epithelial integrity of the colon, as well as the cellular structure of the liver.
Another study with A. mellifera honey demonstrated the stimulating effect of honey on the growth of bifidobacteria and lactobacilli in fermented milk (Macedo et al., 2008). With the addition of sunflower (Helianthus annuus) honey in yogurt, an increase in the values of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus was observed (Sert et al., 2011). While acacia (Acacia) and chestnut (Castanea) honeys had a stimulating effect on the growth of Lactobacillus casei Lc-01 in cows and goats (Sla canac et al., 2011) Pandiyan et al. (2012) demonstrated that A. mellifera honey maintained the level of L. acidophilus above the therapeutic level (10 6 CFU/mL) during 15 days of storage in ice cream. Sla canac et al. (2012) in addition to having observed the proliferation of Bifidobacterium lactis and Bifidobacterium longum during the fermentation of soy (Glycine max) milk, detected an inhibitory activity against L. monocytogenes. Bakr et al. (2015) reported that the addition of 5% fennel (Foeniculum vulgare) honey in bioyogurt increased the count of B. bifidum and Str. thermophilus until the end of the 14-day storage period, in addition to inhibiting the growth of coliforms and molds.
The incorporation of the honey produced M. scutellaris in goat yogurt increased the L. acidophilus La-05 counts until 21 days of storage, maintaining the stability characteristics: color, syneresis, viscosity and water retention capacity, for 7 more days (Machado et al., 2017). Jata ı (Tetragonisca angustula) and Africanized (Apis mellifera) bee honeys supported the viability of probiotic cultures obtained with Lactobacillus acidophilus LA-5 and Bifidobacterium BB-12 in bioyoghurt (Caldeira et al., 2018). And it is due to the fact that the oligosaccharides in honey vary according to their floral origin, that the prebiotic effect of the different honeys is different (da Silva et al., 2016).
Several studies have shown that honey-derived oligosaccharides have similar Prebiotic Index (PI) values to commercial oligosaccharides. Kajiwara et al. (2002) reported duplication times of different species of intestinal bifidobacteria grown in the presence of honey, commercial oligosaccharides and inulin, observing a decrease in replication times for Bifidobacterium breve ATCC 15700 TM with 3.8 ± 0.6 for honey, 2.8 ± 0.2 for fructo-oligosaccharides (FOS), 2.9 ± 0.3 for galacto-oligosaccharides (GOS) and 2.8 ± 0.3 for inulin, while in Bifidobacterium infantis ATCC 15697 TM were obtained 19.5 ± 9.4, 14.7 ± 1.1, 15.8 ± 2.5 and 34.1 ± 3.3, respectively. In addition to demonstrating that the production of lactic and acetic acid by the bifidobacteria studied was increased in a similar way to commercial FOS, GOS and inulin.
Contrarily, Sanz et al. (2005) stated that honey oligosaccharides seem to present a potential prebiotic activity (IP values between 3.38 and 4.24), increasing the bifidobacteria and lactobacillus populations, although not up to the FOS levels (IP of 6.89).

Conclusions
Honey as an ingredient in the food industry, in addition to physicochemical properties, has been shown to act as an antioxidant, mainly for the content of phenolic compounds; as a probiotic, due to the lactic acid bacteria transferred by the bees to the honey; and as a prebiotic, because contains fructose and oligosaccharides. The antibacterial effect is the result of several inherent attributes, such as high sugar concentration, high viscosity and osmotic pressure. These properties which can be altered by the thermal treatments to which the honey will be subjected during the production process. The stingless bees' honey presents high antioxidant and antimicrobial capacity equal to or even better than that of A. mellifera, which together with the increase in its production and the prospect of a wider market for natural products suggest the need to continue researching the composition of this honey. However, there are still few articles published that analyze the aforementioned properties, so this stingless bee honey should stimulate new lines of research to consolidate its consumption and application as an alternative in the food industry.