Influence of stingless bee genus (Scaptotrigona and Melipona) on the mineral content, physicochemical and microbiological properties of honey

Suelen Ávila; Marcelo Lazzarotto; Polyanna Silveira Hornung; Gerson Lopes Teixeira; Vivian Cristina Ito; Marcelo Barba Bellettini; Márcia Regina Beux; Trust Beta; Rosemary Hoffmann Ribani

doi:10.1007/s13197-019-03939-8

. 2019 Jul 23;56(10):4742–4748. doi: 10.1007/s13197-019-03939-8

Influence of stingless bee genus (Scaptotrigona and Melipona) on the mineral content, physicochemical and microbiological properties of honey

Suelen Ávila ^1,^✉, Marcelo Lazzarotto ², Polyanna Silveira Hornung ^1,⁵, Gerson Lopes Teixeira ³, Vivian Cristina Ito ⁴, Marcelo Barba Bellettini ¹, Márcia Regina Beux ¹, Trust Beta ⁵, Rosemary Hoffmann Ribani ¹

PMCID: PMC6801274 PMID: 31686706

Abstract

Stingless bees, important pollinating insects in the tropics, produce honey whose unique quality features differentiate their origin. The feasibility of multivariate data analysis for quality discrimination of stingless bee honey from different genera (Melipona bicolor, quadrifasciata, marginata and Scaptotrigona bipunctata) by mineral content, physicochemical and microbiological properties were investigated. The principal component analysis explained 72.12% of the total variance of the data, and the separation into two main groups in a scatter plot was observed. Group 2 was formed by Scaptotrigona genus, that showed the highest values of pH, ash, and soluble solids. Potassium was the most abundant mineral followed by calcium and sodium for both groups quantified by inductively coupled plasma optical emission spectrometry. This honey has higher acidity and moisture than Apis mellifera honey. Microbiological analyses showed that total aerobic mesophiles ranged between 2.00 and 4.77 log CFU/g. Salmonella spp. was not detected, while the mould and yeast content was above the maximum allowed under the Apis mellifera honey legislation. The evaluated honey samples presented the lactic acid bacteria, which are considered a benefit. The multivariate statistical analysis was efficient in discriminate stingless bee honey, contributing to approaches that can be used for standardization and regulation.

Electronic supplementary material

The online version of this article (10.1007/s13197-019-03939-8) contains supplementary material, which is available to authorized users.

Keywords: Melipona, Scaptotrigona, Brazilian honey, Chemometrics, Principal component analysis, Hierarchical cluster analysis

Introduction

Stingless bees belong to the Hymenoptera family and Meliponinae subfamily. They are placed in one of the four tribes of the Apidae family (Apinini, Bombini, Euglossini and Meliponini) (Michener 2013). Within the Meliponini tribe, attention has been given to Melipona bicolor (Guaraipo), Melipona quadrifasciata (Mandaçaia), Melipona marginata (Manduri) and Scaptotrigona bipunctata (Tubuna). The stingless bees produce honey with an unusual degree of sourness, sweetness, acidity and medicinal value (Chuttong et al. 2016). The chemical composition, flavour, and aroma of honey are strongly associated with its botanical source, geographical area, environmental conditions, bee species involved in its production and storage conditions. Although the production of honey by a stingless bee is lower than the traditional Apis mellifera, the added value for the former species is higher than the latter; prices can be 2–5 times the price of Apis mellifera honey (Ávila et al. 2019).

A serious problem that some producers are facing is that the available official methods for honey quality control have been developed exclusively for Apis mellifera bee’s honey. Due to limited knowledge about stingless bee honey, there are still neither international standards nor regulations created by the authorities for this food (Ávila et al. 2018a). Microbiological, along with physicochemical characteristics, constitute the quality indicators, which describe individual varieties of honey (Manzanares et al. 2011). Analytical techniques such as multivariate analysis and chemometric evaluations have been successfully applied in the quality control of stingless bees honey (SBH), which are able to (Ávila et al. 2018a, b).

The prospects of a broad market regarding natural and organic products, as well as the increased production of SBH, are important for the conservation and maintenance of the flora and native bees (Biluca et al. 2016). In this sense, the evaluation of the composition of honey, including the differences among species with higher production, determining possible conservation strategies and parameters to protect consumers against adulteration to ensure product quality, is necessary. This approach will assist authorities to create standards to evaluate SBH. Thus, in this study, the mineral content, physicochemical and microbiological properties of thirty-two honey samples from two south Brazilian stingless bee’s genera (Scaptotrigona and Melipona) were evaluated, and a method to discriminate SBH using chemometrics tools was obtained.

Materials and methods

Samples

Thirty-two honey samples were collected from four native Brazilian bee species, during the January 2016 in Mandirituba—Paraná (Brazil), according to Ávila et al. (2019).

Physicochemical analyses and microbiological contamination

All the physicochemical parameters (moisture, soluble solids (°Brix), electrical conductivity, colour, pH, total acidity, ash content, hydroxymethylfurfural, reducing sugars and sucrose) were determined in triplicate according to standard methods of the International Honey Commission (IHC 1997) and the Official Methods of Analysis of Association of Official Analytical Chemists—AOAC (2008). Ash content was evalueted by incinerating the samples in a muffle furnace (Quimis, São Paulo, Brazil) at 550 °C. The samples ashes (incinerated samples in a muffle furnace) were solubilized in an acid medium in a 25-mL flask. From the ash solution, the mineral content constituents were determined at wavelengths ranging from 167.019 to 766.491 nm using an inductively coupled plasma optical emission spectrometer (ICP-OES, Varian 720 ES-axial, USA). Twenty-two minerals molybdenum (Mo), nickel (Ni), iron (Fe), magnesium (Mg), manganese (Mn), tin (Sn), vanadium (V), zinc (Zn), phosphorus (P), lead (Pb), copper (Cu), boron (B), calcium (Ca), silver (Ag), chromium (Cr), cobalt (Co), aluminum (Al), barium (Ba), cadmium (Cd), lithium (Li), potassium (K), sodium (Na) were determined in honey samples (n = 32) in triplicate. Water activity (a_w) was determined using an electronic dew-point water activity meter AquaLab 3TE system (Decagon, Washington) at 25 ± 0.02 °C. Aerobic mesophilic bacteria, total coliforms and thermotolerant, lactic acid bacteria (Elliker agar) and Salmonella spp. were determined according to da Silva et al. (2010).

Statistical analysis

The results were presented as a mean ± standard deviation, and the normality was checked by the Kolmogorov–Smirnov test to all variables. Subsequently, the Brown–Forsythe test was carried out to check for homogeneity of all variances (Action Software v 2.5, Estatcamp, Brazil). One-way ANOVA was applied to verify significant differences among samples, and Duncan’s test was used to compare the means when significant differences were detected (P < 0.05). The Pearson correlation coefficients were calculated using the mean values (n = 32) to determine the degree of association between the pairwise variables. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were applied to describe the relationship between the physicochemical properties (n = 9) of honey (n = 32) and the stingless bee’s genera. In PCA, eigenvalues greater than 1.0 were adopted to explain the projection of the samples on the graph and two-dimensional analysis were based on linear correlations. In HCA, the Euclidean metric and Ward’s method were used to suggest groups of similar samples (Zielinski et al. 2014; Ávila et al. 2019).

Results and discussion

Physicochemical analyses

European legislation (EC Directive 2001/110) recommends a minimum of 60 g/100 g of reducing sugars for floral honey. The average reducing sugar in SBH samples was about 54.92 ± 7.18% (w/w). All SBH had more than 45 g/100 g of reducing sugars, but only six showed values above 60 g/100 g in accordance with the international regulations (Codex Alimentarius Commission 2001). The total sugar content averaged 56.32 ± 7.21 g/100 g. The mean percentage of sucrose was 1.40%, but values up to 4.78% were also recorded. The samples under investigation contained lower total sugar content compared to the Apis mellifera honey. The HMF content in SBH ranged from 0.15 to 3.19 mg/kg in agreement with the standard limits for Apis mellifera honey (Codex Alimentarius Commission 2001).

The a_w ranged from 0.77 to 0.91 (Table 1). As a result, a correlation between moisture and water activity (a_w) (Fig. 1a) was observed. Overall, the samples exhibited moisture values ranging between 26 and 40%, higher than the maximum allowed for Apis mellifera, which is 20%. As highlighted in Table 2, due to lower sweetness and higher moisture than the conventional Apis, SBH would require greater levels of caution during harvesting and processing. The acidity of SBH is another parameter which is higher than that exhibited by the Apis. This has been attributed to the enzyme activity over glucose, which has been shown to increase with the sugar dilution in honey (Abadio Finco et al. 2010). All SBH samples recorded electrical conductivity measurements below 0.8 mS/cm, suggesting that these honey samples were of floral origin (Feás et al. 2010). A total of 60% of the evaluated honey were in confirmity with the international regulations for acidity maximum limits (Codex Alimentarius Commission 2001) (Table 1).

Table 1.

Identity and quality parameters of stingless bee honey (SBH) from species of the genus Melipona and Scaptotrigona

Stingless bee species	Sample	Colour (mm Pfund)	Moisture (g/100 g)	Ash (g/100 g)	Electrical conductivity (mS/cm)	pH	Total acidity (meq/kg)	HMF (mg/kg)	Water activity (aw)	°Brix	Reducing sugar (g/100 g)	Sucrose (g/100 g)
Melipona bicolor	MB1	0.67	30.90 ± 0.70^jk	0.09 ± 0.01^ijk	0.26 ± 0.01^r	3.47 ± 0.02^j	34.97 ± 3.08^no	0.19 ± 0.17ⁱ	0.83ⁱ	66.83 ± 0.76^f	56.89 ± 0.42^d	1.97 ± 0.60^efgh
	MB2	6.98	32.28 ± 0.23ⁱ	0.03 ± 0.02^mn	0.23 ± 0.01^s	3.32 ± 0.02^lm	29.43 ± 8.49^op	0.23 ± 0.16^hi	0.84^hi	65.27 ± 0.03^gh	59.72 ± 2.51^c	0.46 ± 0.13^klm
	MB3	0.67	35.58 ± 0.20^e	0.01 ± 0.00ⁿ	0.20 ± 0.01^t	3.35 ± 0.03^l	50.24 ± 1.21^ij	0.31 ± 0.15^ghi	0.88^cd	61.75 ± 0.25^j	77.69 ± 0.86^a	1.11 ± 0.63^ijkl
	MB4	0.30	34.00 ± 0.05^g	0.11 ± 0.02ⁱ	0.23 ± 0.01^s	3.29 ± 0.02^m	35.64 ± 3.50^no	0.87 ± 0.17^cde	0.86^defg	63.17 ± 0.52ⁱ	52.70 ± 0.63^gh	0.38 ± 0.26^lm
	MB5	13.29	32.68 ± 0.32ⁱ	0.02 ± 0.01ⁿ	0.28 ± 0.01^p	3.30 ± 0.02^m	39.80 ± 1.31^mn	–	0.87^def	63.67 ± 0.58ⁱ	48.08 ± 1.03^jk	0.39 ± 0.19^lm
	MB6	23.69	39.94 ± 0.20^ab	0.02 ± 0.02ⁿ	0.32 ± 0.01^l	3.10 ± 0.03^p	63.45 ± 3.05^fg	0.15 ± 0.00ⁱ	0.91^a	57.00 ± 0.25ⁿ	54.32 ± 1.43^defg	4.78 ± 0.16ª
	MB7	46.72	32.94 ± 0.23^ghi	0.09 ± 0.01^ij	0.32 ± 0.01^l	3.23 ± 0.03ⁿ	65.54 ± 7.78^f	–	0.85^ghi	64.67 ± 0.14^h	53.05 ± 0.72^fgh	1.50 ± 0.41^fghij
	MB8	48.58	37.37 ± 0.52^d	0.20 ± 0.01^f	0.31 ± 0.01^l	3.42 ± 0.01^k	58.03 ± 3.01^gh	0.66 ± 0.05^efg	0.90^ab	59.83 ± 0.80^kl	55.55 ± 2.36^def	0.47 ± 0.22^klm
Melipona quadrifasciata	MQ1	0.67	40.07 ± 0.64^a	0.18 ± 0.01^fg	0.29 ± 0.01^op	3.29 ± 0.04^m	45.58 ± 1.21^jklm	1.03 ± 0.04^bcd	0.90^ab	56.75 ± 0.66ⁿ	45.33 ± 1.45^l	0.41 ± 0.13^lm
	MQ2	14.78	35.25 ± 0.23^e	0.11 ± 0.02^hi	0.29 ± 0.01^op	3.09 ± 0.04^p	48.70 ± 3.60^jkl	0.62 ± 0.08^efgh	0.87^def	62.17 ± 0.29^j	55.39 ± 0.92^def	2.88 ± 0.35^cd
	MQ3	23.69	37.17 ± 0.86^d	0.01 ± 0.01ⁿ	0.38 ± 0.01^h	3.08 ± 0.05^p	73.25 ± 0.92^e	0.32 ± 0.21^ghi	0.90^ab	60.50 ± 0.66^k	50.71 ± 0.64^hij	0.33 ± 0.30^lm
	MQ4	28.15	38.82 ± 0.30^bc	0.04 ± 0.00^lmn	0.37 ± 0.01ⁱ	2.97 ± 0.04^q	83.21 ± 7.23^d	0.37 ± 0.18^ghi	0.90^a	58.25 ± 0.25^m	48.56 ± 1.82^ijk	3.95 ± 0.92^b
	MQ5	33.72	35.85 ± 0.23^e	0.11 ± 0.03ⁱ	0.34 ± 0.01^k	3.16 ± 0.01^o	84.66 ± 2.91d	0.75 ± 0.63^def	0.87^cde	61.50 ± 0.25^j	60.00 ± 1.05^c	0.18 ± 0.15^m
	MQ6	69.00	34.07 ± 0.11^fg	0.06 ± 0.01^jklm	0.29 ± 0.01ⁿ	3.22 ± 0.04ⁿ	45.64 ± 2.22^jklm	–	0.86^defg	65.67 ± 0.14^g	50.95 ± 0.35^hi	0.51 ± 0.34^klm
	MQ7	63.80	29.38 ± 0.82^lm	0.01 ± 0.00ⁿ	0.31 ± 0.01^m	3.14 ± 0.03^o	55.80 ± 2.10^hi	–	0.86^defg	68.42 ± 0.95^e	54.64 ± 0.38^defg	0.48 ± 0.24^klm
	MQ8	87.20	35.19 ± 0.73^ef	0.21 ± 0.02^f	0.44 ± 0.01^g	3.32 ± 0.02^lm	101.90 ± 0.57^c	1.34 ± 0.07^b	0.79^jk	63.00 ± 0.50ⁱ	48.07 ± 0.64^jk	0.26 ± 0.12^lm
Melipona marginata	MM1	0.67	32.61 ± 0.71ⁱ	0.15 ± 0.01^gh	0.21 ± 0.01^t	3.43 ± 0.03^k	26.15 ± 0.93^p	0.47 ± 0.08^fghi	0.85^fgh	65.08 ± 0.76^gh	49.82 ± 0.90^ij	1.88 ± 0.88^efghi
	MM2	5.12	38.03 ± 0.30^cd	0.04 ± 0.01^mn	0.29 ± 0.01^op	3.18 ± 0.07^o	49.82 ± 5.87^ijk	–	0.90^ab	59.42 ± 0.38^l	54.91 ± 1.06^defg	2.43 ± 0.84^cde
	MM3	0.67	31.82 ± 0.20^ij	0.11 ± 0.01^hi	0.15 ± 0.01^u	3.57 ± 0.04^gh	18.59 ± 1.47^q	0.80 ± 0.09^def	0.84^hi	65.75 ± 0.25^g	54.98 ± 0.69^defg	2.86 ± 0.29^cd
	MM4	13.67	33.87 ± 0.11^gh	0.12 ± 0.00^hi	0.34 ± 0.01^k	3.51 ± 0.03^ij	46.99 ± 0.87^jklm	–	0.86^defg	63.67 ± 0.14ⁱ	48.61 ± 0.21^ijk	2.25 ± 0.33^def
	MM5	11.07	37.43 ± 0.23^d	0.05 ± 0.01^klmn	0.28 ± 0.01^q	3.17 ± 0.01^o	43.23 ± 1.45^jklm	0.27 ± 0.02^hi	0.89^bc	59.83 ± 0.14^kl	55.60 ± 2.42^def	0.74 ± 0.64^jklm
	MM6	32.98	32.81 ± 0.34^hi	0.02 ± 0.02ⁿ	0.32 ± 0.01^l	3.24 ± 0.03ⁿ	28.88 ± 1.07^op	0.35 ± 0.09^ghi	0.86^efgh	64.75 ± 0.43^h	49.29 ± 0.53^ijk	0.81 ± 0.19^jklm
	MM7	90.54	34.07 ± 0.11^fg	0.33 ± 0.02^d	0.56 ± 0.01^e	3.53 ± 0.01^hi	119.64 ± 1.41^b	1.38 ± 0.09^b	0.86^defg	65.50 ± 0.00ⁱ	56.08 ± 1.04^de	1.33 ± 0.11^hijk
	MM8	152.57	39.81 ± 0.30^ab	0.41 ± 0.03^b	0.66 ± 0.01^a	3.59 ± 0.00^fg	140.36 ± 2.71^a	0.64 ± 0.04^efg	0.90^ab	57.17 ± 0.38ⁿ	46.90 ± 0.46^kl	0.77 ± 0.43^jklm
Scaptotrigona bipunctata	SB1	5.87	26.87 ± 0.40^op	0.08 ± 0.02^ijkl	0.29 ± 0.01^no	3.62 ± 0.02^f	35.75 ± 5.16^no	0.66 ± 0.09^efg	0.79^j	70.92 ± 0.14^b	48.68 ± 0.31^ijk	2.19 ± 0.22^defg
	SB2	14.41	28.39 ± 0.11^mn	0.19 ± 0.01^f	0.38 ± 0.01^h	3.74 ± 0.01^d	42.81 ± 1.29^klm	–	0.80^j	66.50 ± 0.00^f	55.09 ± 2.44^defg	3.14 ± 0.93^c
	SB3	49.32	27.93 ± 0.30^no	0.21 ± 0.00^f	0.32 ± 0.01^l	3.72 ± 0.02^de	41.40 ± 1.13^lmn	1.16 ± 0.16^bc	0.77^k	70.00 ± 0.43^c	55.52 ± 0.53^def	2.48 ± 0.39^cde
	SB4	60.46	26.80 ± 0.11^op	0.55 ± 0.05^a	0.64 ± 0.01^b	4.81 ± 0.01^a	22.70 ± 0.66^pq	3.19 ± 0.61^a	0.78^jk	71.08 ± 0.14^b	60.73 ± 1.64^c	1.06 ± 0.70^ijklm
	SB5	65.66	29.91 ± 0.23^kl	0.28 ± 0.05^e	0.35 ± 0.01^j	3.58 ± 0.02^g	40.66 ± 7.78^mn	1.08 ± 0.11^bcd	0.80^j	67.83 ± 0.14^e	56.08 ± 1.68^de	0.26 ± 0.02^lm
	SB6	93.51	25.88 ± 0.20^p	0.35 ± 0.04^cd	0.61 ± 0.01^d	3.89 ± 0.00^c	63.64 ± 4.78^fg	–	0.77^k	72.08 ± 0.14^a	63.03 ± 1.54^b	0.32 ± 0.02^lm
	SB7	99.09	26.80 ± 0.64^op	0.36 ± 0.02^c	0.61 ± 0.02^c	4.03 ± 0.01^b	46.62 ± 6.65^jklm	–	0.79^j	71.17 ± 0.52^b	54.09 ± 1.17^efg	1.01 ± 0.30^jklm
	SB8	118.03	28.65 ± 0.20^mn	0.42 ± 0.06^b	0.49 ± 0.01^f	3.68 ± 0.01^e	68.27 ± 3.19^ef	1.15 ± 0.08^bc	0.87^def	69.25 ± 0.25^d	76.23 ± 3.25ª	1.39 ± 0.14^ghij
P (Normality)*		–	0.86	0.69	0.09	0.15	0.81	0.66	0.05	0.99	0.14	0.18
P (Brown–Forsythe) **		–	0.55	0.53	0.30	0.86	0.80	0.07	0.54	0.89	0.84	0.66
P (ANOVA)***		–	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

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*Probability values obtained second Kolmogorov–Smirnov test for normality; **Probability values obtained by the Brown–Forsythe test for homogeneity of variances; ***Probability values obtained by one-way ANOVA. Different letters in the same column represent results with statistical difference, according to the Duncan test (P ≤ 0.05)

Fig. 1 — Correlation between a water activity and moisture, and b pH and ash. c Scatter plot of PC1 × PC2 scores of the main sources of variability between Brazilian honey from species of the stingless bees. d Loadings plot of mineral content and physicochemical properties used in the PCA. e Dendrogram obtained for normalised mineral content and physicochemical properties and their respective stingless bee honey

Table 2.

Mean and ranges for stingless bee honey groups and limits for Apis mellifera honey quality parameters

Quality parameters	Mean ± standard deviation (n = 32)	Range (n = 32)	Cluster 1 (n = 24)	Cluster 2 (n = 8)	Limits Apis mellifera honey
Moisture	33.22 ± 0.46	25.88–40.07	35.08^a (29.38–40.07)	27.65^b (25.88–29.91)	Not more than 20 g/100 g*
Total acidity	54.73 ± 3,08	18.59–140.36	57.90 (18.59–140.36)	45.23 (18.59–140.36)	Not more than 50 meq/kg*
Ash	0.13 ± 0.02	0.01–0.55	0.07 (0.01–020)	0.30 (0.19–0.55)	Not more than 0.6 g/100 g*
pH	3.44 ± 0.02	2.97–4.81	3.29 (2.97–3.59)	3.88 (3.58–4.81)	3.3-4.6**
Water activity	0.85 ± 0.04	0.77–0.91	0.87 (0.79–0.91)	0.80 (0.77–0.87)	–
Brix	64.26 ± 4.40	56.75–72.08	62.40 (56.75–68.42)	69.85 (66.50–72.08)	–
Electrical conductivity	355.01 ± 131.70	154.17–660.67	319.90 (154.17–660.67)	460.33 (318.33–635.00)	–
Total mineral	346.84 ± 535.37	37.78–2723.85	359.51 (37.78–2723.85)	308.84 (126.20–670.51)	–
Color	39.86 ± 40.03	0.30–152.57	41.50 (0.30–152.57)	34.9 (30.67–99.09)	–
Reducing sugar	54.92 ± 1.16	45.33–77.69	–	–	Not less than 60 g/100 g*
Sucrose	1.4 ± 0.36	0.18–4.78	–	–	Not more than 5 g/100 g*
HMF	0.56 ± 0.29	0.00–3.19	–	–	Not more than 60 mg/kg*

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*Codex Alimentarius Commission (2001); **Brasil (1985)

The °Brix in the SBH was lower than that of Apis mellifera honey due to the higher moisture content. The samples showed an average pH of 3.44 ± 0.35 (Table 1). The pH values were in agreement with the Brazilian legislation (Brasil, 1985), which establishes a range of 3.3–4.6 for Apis mellifera honey (Table 2). A correlation between pH and mineral content from SBH samples was observed (Fig. 1b). Honey can be classified as blossom honey (nectar of plants) or honeydew honey (secretions of living parts of plants or excretions of plant-sucking insects) (Pita-Calvo and Vazquez 2017). In that context, all the SBH showed less than 0.60% ash content, which is recommended for blossom honey (Codex Alimentarius Commission 2001). Low ash content is a characteristic of light-coloured honey (Khalafi et al. 2015). The Pfund colour evaluation showed that 28.1% of the samples were classified as water white, 15.6% extra white, 15.6% white, 12.5% light amber, 12.5% amber, 9.4% extra light amber and 6.3% dark amber.

Potassium (K) was the most abundant element in all analyzed samples (Online Resource 1). It is considered the most important mineral (quantitatively) in the Apis mellifera honey (3.90–491.40 mg/100 g), representing about 50% of the total mineral content (Czipa et al. 2015). The second most abundant mineral was calcium (Ca) in all SBH samples. The other two minerals frequently present in SBH were Mg and Na. Mn, a heavy metal considered as an essential micronutrient was present in SBH. According to the number of minerals found, ingestion of a spoon (approximately 20 g) of SBH would supply: 4.55 mg of K, 0.64 mg of Ca, 0.59 mg of Na, 0.31 mg of Mg, 47.29 µg of Zn, 40.24 µg of Mn, 22.69 µg of Fe, 13.44 µg of Cr, 4.21 µg of Cu and 0.46 µg of V. According to the reference daily intake for adults, 2 tablespoons of SBH would correspond to the intake of 0.12% of the recommended for Ca, 0.24% of Mg, 1.14% of Zn, 3.5% of Mn, 3.2 of Fe, 76% Cr and 0.02% Cu.

Few samples of MB2, MB4 and MQ7 showed traces of tin (Sn) with values ranging from 0.02 to 0.06 mg/kg. The samples MQ8, SB2 and SB7 had values ranging from 0.01 to 0.02 mg/kg for cobalt. Pb, Cd, and Al are classified as environmental micro contaminants and are not necessary to the living organism in any quantity. MB3, MB7 and SB6 samples had traces of cadmium in levels ranging from 0.01 to 0.04 mg/kg.

In order to understand the data set, interrelationships, and the differences between the bee species groups, principal component analysis (PCA) was applied to the data on moisture, a_w, electrical conductivity, total acidity, °BRIX, pH, colour, ash and total mineral of Brazilian SBH (Tables 1, 2). The PCA explained 72.12% of total data variance (Fig. 1c). In the scores plot, the honey samples were successfully separated into two groups, Scaptotrigona and Melipona types. The Melipona genus group (Fig. 1c, d, right side) showed higher levels of moisture, water activity, colour, electrical conductivity, total acidity and total mineral in the loading plot. The Scaptotrigona genus group (Fig. 1c, d, left side), showed higher levels of °Brix, pH and ash. Hierarchical Cluster Analysis (HCA) was performed to verify the grouping of the samples according to their mineral content and physicochemical properties in the dendrogram shown in Fig. 1e. Two large clusters can be observed in the dendrogram, which separates the samples from species of Melipona genus (Cluster 1) and the samples from species of Scaptotrigona genus (Cluster 2). These findings corroborated the results obtained by PCA.

Microbiological evaluation

In natura honey usually present microorganisms that survive in conditions of concentrated sugars, low a_w, acidity conditions and antimicrobial properties, influencing their quality and safety for consumption. Due to their low pH and high acidity, honey from stingless bees create an inhospitable environment for microorganisms, especially the pathogens, therefore, they are considered a food with low risk for consumers (Estevinho et al. 2012). The microbiota that can grow in pH lower than 4.5 comprises the moulds and yeasts besides the lactic bacteria. Yeast and moulds were detected in SBH in high counts, with a mean value of 3.4 ± 0.61 log CFU/g. A value lower than 3 log CFU/g has been reported for yeast and moulds in Apis mellifera honey (Online Resource 2).

The bacterial presence could indicate contamination during handling, processing, and storage, and they can be avoided by the application of good manufacturing practices (Różańska 2011). However, there is an abundant flora of lactic acid bacteria (LAB) in honey that is generally recognized as benefic bacteria (GRAS status) and play an essential role in food fermentation and preservation. The LABs in the SBH ranged from 1.24 to 5.82 log CFU/g, suggesting the presence of a natural preservative. LAB improves the hygienic quality and safety of food by inhibiting the competing flora, which includes many spoilage and pathogenic bacteria. It also produces organic acids (mainly lactic acid) resulting in a lower pH as well as several antimicrobial compounds (Cintas et al. 2001).

Total aerobic mesophile counts ranged from 2.00 to 4.77 log CFU/g. Concerning safety, Salmonella sp. was not detected in any of 32 samples. For the coliform group at 35 °C, 78% of the analyzed honey samples had < 3 MPN/g, and 94% showed no coliforms at 45 °C. Thus, only two samples of the species Scaptotrigona bipunctata had thermotolerant coliforms. These bees may have an unhygienic habit when collecting material for the construction of the hive (Souza et al. 2009). This could also be attributed to some anthropic contamination that may be occurred during the collection of honey or even when transporting the samples, since those microorganisms were detected in less than 6% of the SBH samples, suggesting that they are not naturally present in the honey.

Conclusion

The honey collected from South Brazilian native stingless bees have a floral origin and are derived from nectars that show both low conductivity and ash content. Distinct from the Apis mellifera, the stingless bee honey (SBH) evaluated herein showed an acid character and hygroscopicity due to their slightly lower level of total carbohydrates and high values in moisture and water activity. Besides, SBH can be considered a good source of minerals, particularly K and Ca. This study also showed that the bee species of different genus impact the honey properties. The multivariate statistical analysis was effective to discriminate and to classify the SBH groups. Using the chemometric approach, Principal Component Analysis (PCA), which explained 72.12% of the data variability, was successfully able to distinguish the honey into the Scaptotrigona and Meliponina genera. Two clusters suggested by the Hierarchical Cluster Analysis corroborated with PCA, in which cluster 2 was formed by Scaptotrigona genus, that showed the highest values of pH, ash, and °BRIX. To create standards and to assure suitable treatment to reduce SBH microbiota, it is necessary to consider the significant variance that exists between honey production of different stingless bee’s genera. The increasing commercial production of these honey require official analytical methodologies and standards to be established aiming to help both producers to preserve sensorial and nutritional characteristics and analysts in the evaluation and prevention of adulteration of SBH.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 131 kb)^{(131KB, doc)}

Supplementary material 2 (DOC 80 kb)^{(80KB, doc)}

Acknowledgements

The authors gratefully acknowledge Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-CAPES-Brazil, EMBRAPA Florestas, and Conselho Nacional de Desenvolvimento Científico e Tecnológico-CNPq (306930/2016-1) for the financial resources provided. We also thank the Associação de Meliponicultores de Mandirituba (Amamel) for kindly supplying the honey samples.

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interest to declare.

Footnotes

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Contributor Information

Suelen Ávila, Phone: +55-41-33613232, Email: suelenavila@gmail.com, Email: suelenavila@ufpr.br.

Marcelo Lazzarotto, Email: marcelo.lazzarotto@embrapa.br.

Polyanna Silveira Hornung, Email: polyanna14silveira@gmail.com.

Gerson Lopes Teixeira, Email: gerson775@gmail.com.

Vivian Cristina Ito, Email: vivianito@gmail.com.

Marcelo Barba Bellettini, Email: marcelobeletini@yahoo.com.br.

Márcia Regina Beux, Email: beuxmarcia@gmail.com.

Trust Beta, Email: Trust.Beta@umanitoba.ca.

Rosemary Hoffmann Ribani, Email: roseribani@gmail.com.

References

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Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supplementary material 1 (DOC 131 kb)^{(131KB, doc)}

Supplementary material 2 (DOC 80 kb)^{(80KB, doc)}

[CR1] Abadio Finco FDB, Moura LL, Silva IG. Propriedades físicas e químicas do mel de Apis mellifera L. Cienc e Tecnol Aliment. 2010;30:706–712. doi: 10.1590/S0101-20612010000300022. [DOI] [Google Scholar]

[CR2] AOAC (2008) Association of Official Analytical Chemists. Official methods of analysis of AOAC international

[CR3] Ávila S, Beux MR, Ribani RH, Zambiazi RC. Stingless bee honey: quality parameters, bioactive compounds, health-promotion properties and modification detection strategies. Trends Food Sci Technol. 2018;81:37–50. doi: 10.1016/j.tifs.2018.09.002. [DOI] [Google Scholar]

[CR4] Ávila S, Hornung PS, Teixeira GL, Beux MR, Lazzarotto M, Ribani RH. A chemometric approach for moisture control in stingless bee honey using near infrared spectroscopy. J Near Infrared Spectrosc. 2018;26:379–388. doi: 10.1177/0967033518805254. [DOI] [Google Scholar]

[CR5] Ávila S, Hornung PS, Teixeira GL, Malunga LN, Apea-Bah FB, Beux MR, Beta T, Ribani RH. Bioactive compounds and biological properties of Brazilian stingless bee honey have a strong relationship with the pollen floral origin. Food Res Int. 2019;123:1–10. doi: 10.1016/j.foodres.2019.01.068. [DOI] [PubMed] [Google Scholar]

[CR6] Biluca FC, Braghini F, Gonzaga LV, Costa ACO, Fett R. Physicochemical profiles, minerals and bioactive compounds of stingless bee honey (Meliponinae) J Food Compos Anal. 2016;50:61–69. doi: 10.1016/j.jfca.2016.05.007. [DOI] [Google Scholar]

[CR7] Brasil (1985) Portaria no 6, de 25 de julho de 1985. Normas Higiênico-Sanitárias e Tecnológicas para Mel, Cera de Abelhas e Derivados. Minist da Agric Secr Inspeção Prod Anim

[CR8] Chuttong B, Chanbang Y, Sringarm K, Burgett M. Physicochemical profiles of stingless bee (Apidae: Meliponini) honey from South East Asia (Thailand) Food Chem. 2016;192:149–155. doi: 10.1016/j.foodchem.2015.06.089. [DOI] [PubMed] [Google Scholar]

[CR9] Cintas LM, Casaus MP, Herranz C, Nes IF, Hernandez PE. Review: Bacteriocins of lactic acid bacteria. Rev Agaroquimica Tecnol Aliment. 2001;7:281–305. doi: 10.1106/R8DE-P6HU-CLXP-5RYT. [DOI] [Google Scholar]

[CR10] Codex Alimentarius Commission Codex Alimentarius Commission Standards. Codex Stan. 2001;12–1981:1–8. [Google Scholar]

[CR11] Czipa N, Andrási D, Kovács B. Determination of essential and toxic elements in Hungarian honeys. Food Chem. 2015;175:536–542. doi: 10.1016/j.foodchem.2014.12.018. [DOI] [PubMed] [Google Scholar]

[CR12] da Silva N, Christina V, Ferraz N, Hiromi M, Francisco R, Abeliar R. Manual de Métodos de Análise Microbiológica de Alimentos e Água. 4. Sao Paulo: Editora Blucher; 2010. [Google Scholar]

[CR13] Estevinho LM, Feás X, Seijas JA, Pilar Vázquez-Tato M. Organic honey from Trás-Os-Montes region (Portugal): chemical, palynological, microbiological and bioactive compounds characterization. Food Chem Toxicol. 2012;50:258–264. doi: 10.1016/j.fct.2011.10.034. [DOI] [PubMed] [Google Scholar]

[CR14] Feás X, Pires J, Estevinho ML, Iglesias A, de Araujo JPP. Palynological and physicochemical data characterisation of honeys produced in the Entre-Douro e Minho region of Portugal. Int J Food Sci Technol. 2010;45:1255–1262. doi: 10.1111/j.1365-2621.2010.02268.x. [DOI] [Google Scholar]

[CR15] IHC (1997) Harmonized methods of the International Honey Commision. In: IHC responsible for the methods: Stefan. Switzerland: Swiss Bee Research Centre FAM, Liebefeld (CH-3003), pp 10–16

[CR16] Khalafi R, Goli SAH, Isfahani MB. Characterization and classification of several monofloral Iranian honeys based on physicochemical properties and antioxidant activity. Int J Food Prop. 2015;19:1–32. doi: 10.1080/10942912.2015.1055360. [DOI] [Google Scholar]

[CR17] Manzanares AB, García ZH, Galdón BR, Rodríguez ER, Romero CD. Differentiation of blossom and honeydew honeys using multivariate analysis on the physicochemical parameters and sugar composition. Food Chem. 2011;126:664–672. doi: 10.1016/j.foodchem.2010.11.003. [DOI] [Google Scholar]

[CR18] Michener CD. Pot-honey. New York: Springer; 2013. [Google Scholar]

[CR19] Pita-Calvo C, Vazquez M. Differences between honeydew and blossom honeys: a review. Trends Food Sci Technol. 2017;59:79–87. doi: 10.1016/j.tifs.2016.11.015. [DOI] [Google Scholar]

[CR20] Różańska H. Microbiological quality of polish honey. Bull Vet Inst Pulawy. 2011;55:443–445. [Google Scholar]

[CR21] Souza B, Marchini L, Tadeu C, Oda-souza M, Alfredo C, Carvalho M, Alves R. Avaliação microbiológica de amostras de mel de trigoníneos (Apidae: Trigonini) do Estado da Bahia. Cienc e tenologia Aliment Campinas. 2009;29:798–802. doi: 10.1590/S0101-20612009000400015. [DOI] [Google Scholar]

[CR22] Zielinski AAF, Ávila S, Ito V, Nogueira A, Wosiacki G, Haminiuk CWI. The association between chromaticity, phenolics, carotenoids, and in vitro antioxidant activity of frozen fruit pulp in Brazil: an application of chemometrics. J Food Sci. 2014;79:C510–C516. doi: 10.1111/1750-3841.12389. [DOI] [PubMed] [Google Scholar]

PERMALINK

Influence of stingless bee genus (Scaptotrigona and Melipona) on the mineral content, physicochemical and microbiological properties of honey

Suelen Ávila

Marcelo Lazzarotto

Polyanna Silveira Hornung

Gerson Lopes Teixeira

Vivian Cristina Ito

Marcelo Barba Bellettini

Márcia Regina Beux

Trust Beta

Rosemary Hoffmann Ribani

Abstract

Electronic supplementary material

Introduction

Materials and methods

Samples

Physicochemical analyses and microbiological contamination

Statistical analysis

Results and discussion

Physicochemical analyses

Table 1.

Fig. 1.

Table 2.

Microbiological evaluation

Conclusion

Electronic supplementary material

Acknowledgements

Compliance with ethical standards

Conflict of interest

Footnotes

Contributor Information

References

Associated Data

Supplementary Materials

ACTIONS

PERMALINK

RESOURCES

Cite

Add to Collections

PERMALINK

Influence of stingless bee genus (Scaptotrigona and Melipona) on the mineral content, physicochemical and microbiological properties of honey

Suelen Ávila

Marcelo Lazzarotto

Polyanna Silveira Hornung

Gerson Lopes Teixeira

Vivian Cristina Ito

Marcelo Barba Bellettini

Márcia Regina Beux

Trust Beta

Rosemary Hoffmann Ribani

Abstract

Electronic supplementary material

Introduction

Materials and methods

Samples

Physicochemical analyses and microbiological contamination

Statistical analysis

Results and discussion

Physicochemical analyses

Table 1.

Fig. 1.

Table 2.

Microbiological evaluation

Conclusion

Electronic supplementary material

Acknowledgements

Compliance with ethical standards

Conflict of interest

Footnotes

Contributor Information

References

Associated Data

Supplementary Materials

ACTIONS

PERMALINK

RESOURCES

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