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Contribution to the palynological, physicochemical and organoleptic characterization of Mulinum spinosum (Apiaceae) honeys from Patagonia, Argentina/Contribucion a la caracterizacion palinologica, fisicoquimica y organoleptica de las mieles de Mulinum spinosum (Apiaceae) de Patagonia, Argentina/Contribuicao a caracterizado palinologica, fisico-quimica e organoleptica dos meis de Mulinum spinosum (Apiaceae) de Patagonia, Argentina.

Introduction

Honey is a nutritious food produced by bees, mainly Apis mellifera L., from nectar or exudations excreted by some plant-sucking insects, gathered, modified and stored in honey combs (Codex Alimentarius Commission, 2001). The composition and properties of honey depend on the botanical origin of the nectar or secretion used. If the nectar from which the honey is derived is gathered mainly from flowers of one specific plant species, the product is called unifloral (White, 2005). These honeys are characterized by its particular pollen content and by their organoleptic, physical and chemical properties (Accorti et al., 1986; Bogdanov et al., 2004; Persano Oddo et al., 2004).

The interest in the production of unifloral honey is due to higher consumer preference, as well as the therapeutic or technological use of certain honey types (Ruoff et al., 2006). This recent interest increases the demand for reliable determination of the botanical origin, based primarily on pollen content. Although unifloral honeys have defined properties, flora from different geographical regions can result in variations. Therefore, it is important to characterize honeys by their botanical as well as their geographical origin, according to palynological analysis (Von der Ohe et al., 2004).

Botanical characterization also requires physical and chemical determinations, which provide a useful complement to palynological analysis. The parameters used in the physicochemical examination are: color, electrical conductivity, moisture, hydroxymethyl-furfural (HMF), optical activity, acidity, pH, sugar composition and enzymatic activity (Bogdanov et al., 2004).

Moreover, sensory analysis represents a complement for determining the botanical origin of honey, especially for those characteristics that cannot be studied by analytical methods (Piana et al., 2004; Von der Ohe et al., 2004).

The market demand for beekeeping products of differentiated origins, and the interest of beekeepers in Argentina to increase the added value of them, have led to a large number of studies aimed to determine botanical and geographical origin of honeys from different regions of Argentina (Telleria, 1988; Andrada and Telleria, 2002; Fagundez and Caccavari, 2006; Forcone, 2008; Forcone et al., 2009).

The advance of melisso-palynological studies in different areas of Argentina, particularly in those regions with an abundant native flora, rich in endemic species, has allowed the detection of new types of unifloral honeys. In the Patagonia region were detected 18 types of these honeys, two of whom are from native plants (Telleria and Forcone, 2000; Forcone, 2008; Forcone et al., 2009). These findings raise the need to characterize and determine in each case the pollen percentages established for the designation of unifloral.

Unifloral honey from Mulinum spinosum (Cav.) Pers. (Apiaceae) comes from a subshrub commonly known as 'neneo' which is an Argentine-Chilean endemic species (Zuloaga et al., 2008). The production area of Mulinum honey (Figure 1) is in the extra-Andean Patagonia, in the northwest of Santa Cruz province (Forcone. et al, 2009). The region extends from the sub-Andean foothills and descends eastwards to the Atlantic Ocean; its landscape consists mainly of plateaus with the shrub steppe as physiognomic predominant type, being the most southern apicultural area of Argentina. It is comprised in a transition region between the Patagonian Province and grass steppes of the Subantarctic Province (Roig, 1998). Among the most abundant species stand out: Senecio patagonicus Hook & Arn., Mulinum spinosum (Cav.) Pers., Colliguaja integerrima Gillies et Hook, Adesmia boronioides Hook., Berberis heterophylla Juss., Schinus marchandii Barkley and Acaena magellanica (Lam.) Vahl (Roig, 1998).

The objective of the present study was to characterize unifloral honey from M. spinosum from the palynological, physicochemical and organoleptic points of view, to broaden knowledge of unifloral honey types produced in Argentina and the world.

Materials and Methods

Sixteen honey samples produced by Apis mellifera and classified as unifloral from Mulinum spinosum by the Palynology Laboratory of the Universidad Nacional de la Patagonia San Juan Bosco (UNPSJB), were studied. The samples were collected directly from beekeepers in northwest of Santa Cruz Province, Argentina (46.5[degrees]S, between 70.5[degrees] and 71.7[degrees]W) (Figure 1) during the period 2005-2011 and kept at 4[degrees]C. Pollen samples were obtained by centrifugation of the honey as described below.

Pollen analysis

For the pollen qualitative analysis the methods of Louveaux et al. (1978) were followed, slightly modified. Twenty grams of honey were dissolved in 100ml of distilled water, centrifuged at 1500g (3000rpm) during 10min at at room temperature, washed and acetolysed (Erdtman, 1960). The pollen sediment was mounted on glycerine-gelatin and sealed with paraffin. To determine the relative frequency (percentage of each pollen type in the pollen content of a sample) 500 pollen grains were counted in each case. Pollen types were classified into four categories: predominant pollen (>45% of the total number of pollen grains); secondary pollen (16-45%); important minor pollen (3-15%) and minor pollen (<3%). When only one pollen type represented >45% of the total number of pollen grains, the sample was classified as unifloral honey (Louveaux et al., 1978).

Pollen types were identified by comparing them with a reference pollen collection made from plants from the area surrounding the beehives. The pollen atlases of Heusser (1971) and Markgraf and D'Antoni (1978) were consulted. The reference collection was donated to the Palynotheque of the UNPSJB (Trelew campus). The herbarium specimens were deposited in the Trelew Herbarium (HTW).

Quantitative pollen analysis included the addition of tablets of Lycopodium clavatum L. spores (Stockmarr, 1971; Moar, 1985). Ten grams of honey were dissolved in 40ml of distilled water, and two tablets of L. clavatum spores (each containing 12000 [+ or -] 200 spores) were dissolved in 5ml of 5% hydrochloric acid and added. The sediment was concentrated by repeated centrifuging at 1500g (3000rpm), using a 10ml centrifuge tube. The centrifugation was continued until all the sediment was included in one tube. The sediment, without any chemical treatment, was mounted on glycerine-gelatin, and sealed with paraffin. Spores, pollen and honeydew were counted until 500 pollen grains were reached in each case.

Pollen concentration (NPG; number of pollen grains in 10g of honey) was calculated by applying the formula: (pollen counted/L. clavatum spores counted) x spores added. Samples were distributed into classes according to the pollen grain content of 10g honey (Maurizio, 1939) as: Group I (<20,000); Group II 20,000-100,000); Group III (100,000-500,000); Group IV (500,000-1,000,000); and Group V (>1,000,000). The honeydew index (Louveaux et al., 1978) was calculated as HDE/P (ratio of honeydew elements--HDE--to pollen grains of nectariferous plants--P).

Organoleptic properties

The method presented by Piana et al. (2004) was used in the analyses of organoleptic properties. The method is based on the evaluation of the olfactory-gustatory characteristics of honey to identify sensory stimuli on the basis of previously memorized standards.

Physicochemical analysis

The physicochemical analysis performed included color, moisture, hydroxymethylfurfural (HMF), pH, free acidity, ashes, electrical conductivity, diastase activity, total polyphenols and sugars determinations.

Color was determined by the measurement of absorbance at 635nm of 10g of honey diluted in distilled water to a volume of 20ml (Bianchi, 1990). Additional measurements were also carried out by optical comparison with a Lovibond[TM] tintometer using a Pfund scale (Aubert and Gonnet, 1983). Moisture was determined with an Abbe[TM] refractometer at 20[degrees]C by obtaining the corresponding value of moisture percentage from the Chataway table (AOAC, 1999). Acidity was determined by titration with 0.10M sodium hydroxide (NaOH) until pH reached 8.3 (AOAC, 1999). HMF was measured by spectroscopy (White, 1979). Measurement of electrical conductivity were conducted in a honey solution containing 20% of honey dry matter in 100ml distilled water (Bogdanov et al., 1999). The method of Schade modified by White (White, 1979) was used to determine the diastase activity and its results expressed as ml of 1[degrees]% starch hydrolyzed by the enzyme in 1g of honey in 1h (Codex Alimentarius Commission, 2001).

The levels of polyphenol compounds in the honey samples were estimated with spectrophotometric determination using a modified Folin-Ciocalteu method (Singleton et al., 1999). Briefly, 30ul of honey sample (0.1g x [ml.sup.-1]) was mixed with 150 [micro]l Folin-Ciocalteu's phenol reagent (0.2N). After 2min, 450ul sodium carbonate solution (0.2 g x [ml.sup.-1]) was added to the mixture. The reaction was kept in the dark for 120min, after which the absorbance was read at 765nm by a SP-300 Plus UVVIS spectrophotometer (Optima, Tokio, Japan). Gallic acid was used to calculate the standard curve. Phenolic compound levels were measured in triplicate. The results were expressed as mg of gallic acid equivalent (GAE)/100g honey.

Sugars contents were determined using HPLC according to IHC (Bogdanov, 2002) on a high pressure liquid chromatograph (Agilent, Series 1100, Germany) equipped with binary pumps (G1316A), a termostated autosampler (G1329A), a termostated column oven (G1316A) and a refractive index detector (RID) (G1362A), combined with Agilent Chem Station IL 50508 software. The column was Zorbax N[H.sub.2], 4.6 x 250mm, particle size of 5 [micro]m.

The amount of sample injected onto column was 5[micro]l. The separation was conducted at a temperature of 35[degrees]C with the mobile phase acetonitrile:water (83:17) at a flow rate of 0.65ml x [min.sup.-1]. For the qualification and quantification of the saccharides, the HPLC chromatograms of the samples were compared to those of commercial standards of fructose, glucose, saccharose, turanose, maltose, trehalose, erlose, melezitose, raffinose, maltotriose and maltotetrose.

All physicochemical results were compared with International Regulatory Standards (Bogdanov et al, 1999). Means and standard deviations were calculated by using the software Microsoft[R] Excel 2007.

Results

Pollen analysis

Qualitative analysis. The relative frequency of Mulinun spinosum pollen ranged from 52% to 87% (Table I). Forty six morphological types were determined in the accompanying pollen: 39 from insect-pollinated plants and seven from wind-pollinated plants. The most frequent entomophilous types were: Senecio spp. (Asteraceae), Brassicaceae and Melilotus spp. (Fabaceae); these types were found in 100% of the samples (Table I; Figure 2). Other frequent types were: Acaena spp. (RosaceaeJ, Astereae, Convolvulus arvensis (Convolvulaceae), Medicago sativa (Fabaceae) and Trifolium spp. (Fabaceae; found in 8090% of the samples), Taraxacum officinale (Asteraceae) and Rosaceae (50-60% of the samples). From these types Brassicaceae, Melilotus sp., Acaena spp. and Rosaceae were classified as secondary pollen. Senecio spp., M. sativa and Trifolium spp. were found as important minor pollen together with the tree first mentioned types of pollen, while the remaining types correspond to minor pollen. The most frequent anemophilous types were Plantago spp. and Poaceae (Table I).

Quantitative analysis. All the samples showed moderate pollinic richness. The values of pollen grains per 10g of honey were between 22,220 and 141,425 (Table II), with an average of 71,870.75. Of the samples, 62% belonged to Maurizio's (1939) Group II. The remaining samples were classified in Group III, most of them in the lower limit of that category.

Honeydew indicators were scarce or absent. The HDE/P ratio was <1 in all the samples (Table II).

Sensory analysis

Mulinum spinosum (Cav.) Pers. honey showed light color intensity and amber color tones. Its flavor, sweet and warm, resembled caramel, according to Piana et al. (2004). The honey was homogeneous and presented a creamy consistence with fine crystals.

Physicochemical analysis

All the honey samples presented very low values of HMF, with a maximum value of 4.5mg x [kg.sup.-1]. Water content ranged from 11.4 to 17.1% with an average value of 15.19%. Ash content and electrical conductivity were in agreement with international standards, with a mean value of 0.15% w/w and 0.41mS x [cm.sup.-1], respectively (Table III).

Values of free acidity were low (mean of 20.2mg x [kg.sup.-1]). The average pH value was 4.4 and ranged between 3.7 and 5.3.

With regard to color (Table III), 75% of the samples analyzed ranged mainly between extra light amber (34-45mm Pfund) and light amber (61.38-0.4mm Pfund). Only 12.5% were amber (>94mm Pfund) and 12.5% were white (<34mm Pfund).

The diastase activity recorded for the honey ranged between 5.3 and 30.5 DN with an average of 12.62. Most of the samples showed values between 9 and 19 DN. Only one sample showed a value of 30.5 DN.

M. spinosum honey (Table IV) was rich in fructose and its fructose/glucose ratio was 1.3 and glucose/water ratio 1.9. The values of fructose and glucose were in the range 36.3-40.4g/100g and 24.2-37.4g/100g, respectively. All the samples contained the dissacharides turanose (mean value 2g/100g) and trehalose (mean 0.8g/100g). In addition, the oligosaccharide melezitose was present in six samples and erlose in four samples. Maltotriose was recorded in two samples while raffinose was in only one case. Saccharose, maltose and maltotetraose were not detected.

The total amount of phenolic compounds ranged from 113 mg GAE 100 x [g.sup.-1] to 190 mg GAE 100 [g.sup.-1] with an average value of 143.1 mg GAE 100 [g.sup.-1] (Table III).

Discussion

In general, a honey is considered as coming predominantly from a given botanical origin (unifloral honey) if the relative frequency of pollen of that taxon exceeds 45%, and if the ratio of the number of honeydew elements (HDE) to that of pollen grain does not exceed three (Von der Ohe et al., 2004). The percentages of the Mulinum spinosum pollen reported in the present work (52-87%) greatly exceeds the minimum value established for classifying a honey as unifloral, and in all cases the ratio honeydew elements/pollen grains does not exceed 1[degrees]%. Moreover, the obtained results show a moderate pollen concentration, indicating that pollen from M. spinosum in the pollen content of honey is neither over-represented nor under-represented (Louveaux et al., 1978; Von der Ohe et al., 2004). These data together with sensory analysis allows inferring that the studied honeys are M. spinosum unifloral.

In these honeys, accompanying pollen comes from fodder crops, fruit trees, weed and native plants. The exotic secondary pollens with greater frequency of occurrence in the samples are: Brassicaceae, Melilotus sp. (Fabaceae) and Rosaceae, the latter mainly represented by fruit crops.

The most frequently native pollen come from Acaena spp. (Rosaceae), Astereae and Senecio spp. (Asteraceae); other frequently native pollens are Adesmia spp., Ochetophila trinervis and Phacelia secunda. All these taxa are present in the herbaceous shrub-steppe that characterizes the western district of the Patagonian Province.

Pollen of wind pollinated plants is abundant in M. spinosum honeys, most of the anemophilous pollen being from Plantago sp. The abundance of anemophilous pollen differentiates extra Andean Patagonian honeys from those originated in other Argentinean areas, where pollen of wind pollinated plants is scarce (Telleria, 1988; Fagundez and Caccavari, 2006).

The physicochemical characteristics studied in M. spinosum honeys could be related to the geographical origin and the climate of the production area, with annual precipitation ~200mm and an average annual temperature of 8[degrees]C (Forcone et al, 2009). The low values of moisture and HMF measured in the samples could be attributed to the extreme dryness of the region of origin. These results agree with those found in honeys of Chubut Province (Argentinean Patagonia) with similar climate conditions (Aloisi, 2010).

Regarding color, honeys from M. spinosum present light tones, varying from white to amber. The clearest samples showed higher percentages of dominant pollen than those that were darker. These results indicate that in these honeys the lightest tones would be related to a greater contribution of nectar from M. spinosum.

The characteristics of pollen content and the low values of conductivity indicate that the analyzed honeys originate mainly from nectar. The conductivity values are in the range establish for nectar honeys (Bogdanov et al., 1999).

Eight carbohydrates were identified, including two monosaccharides, two disaccharides and four oligosaccharides. The monosaccharides glucose and fructose were present in every sample and were the main sugars in all of them. The prevalence of these monosaccharides agrees with the sugar profile characteristic of blossom honeys (Bogdanov et al., 2004). The glucose levels detected in M. spinosum honeys correspond to the values determined for unifloral honeys (Persano Oddo and Piro, 2004).

Polyphenols are an important group of compounds with respect to the appearance and the functional properties of honey (Bogdanov et al., 2008). These phytochemical constituents are considered to be very important in the assessment of honey flavor quality and, hence, the overall quality of honey and its botanical origin. Different types and quantities of phenolic compounds may also vary according to the floral origin (Sant'Ana et al., 2012). The total amounts of polyphenols registered in the studied samples are within the values determined for others unifloral honeys (Bogdanov et al., 2008).

Values obtained for M. spinosum honey are consistent with the values for other Apiaceae honeys studied in other latitudes, specifically in Morocco (Ammi visnaga, Eryngium campestre, Ridolfia segetum; Terrab et al., 2003). Both Apiaceae Moroccan honeys and M. spinosum honey are characterized by clear tones ranging from white to amber and a moderate pollen concentration corresponding to Groups II and III of Maurizio's classification. Both types present similar pH, free acidity and electrical conductivity. However, the lower levels of HMF and water content detected in the Mulinum honeys can differentiate them from the Apiacea honeys produced in Morocco.

Received: 08/03/2013. Modified: 07/08/2013. Accepted: 22/08/2013.

Conclusions

The studied honeys were found to meet the requirements of the international honey standards and the results contribute to the knowledge of unifloral honeys produced in Argentina and in the world.

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Forcone A, Aloisi P, Munoz M (2009) Palynological and physicochemical characterization of honey from north-west of Santa Cruz (Argentina). Grana 48: 70-79.

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Markgraf V, D'Antoni H (1978) Pollen flora of Argentina. University Arizona Press. Tucson, AZ, USA. 208 pp.

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Persano Oddo L, Piana L, Bogdanov S, Bentabol A, Gotsiou P, Kerkvliet J, Martin P, Morlot M, Ortiz Valbuena A, Ruoff K, Von Der Ohe K (2004) Botanical species giving unifloral honey in Europe. Apidologie 35: 82-93.

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Pia V. Aloisi. Biologist, Universidad de Buenos Aires, Argentina. Magister in Food Technology, Universita degli Studi di Parma, Italia. Professor, Universidad Nacional de la Patagonia San Juan Bosco (UNPSJB), Argentina. Address: Laboratorio de Palinologia y Control de Calidad de Mieles y Productos Apicolas, Facultad de Ciencias Naturales, UNPSJB. Roca 115, 9100 Trelew, Chubut, Argentina. email: piaaloisi@gmail.com

Alicia E. Forcone. Biologist, Universidad Nacional de Cordoba (UNC), Argentina. Doctor in Biology, Universidad Nacional del Sur, Argentina. Professor, UNC and UNPSJB, Argentina. email: aeforcone@ yahoo.com.ar

Marisa Amadei. Biochemist, Universidad Nacional de la Plata, Argentina. Technical Director and Quality Manager, Laboratorios NEXCO S.A., Buenos Aires, Argentina. email: m.amadei@nexco-sa. com.ar

TABLE I

FREQUENCY CLASES

Family                                       Samples

                                        1    2    3    4

Anacardiaceae     Schinus marchandii
                  F. A. Barkley *

Apiaceae          Conium maculatum L.

                  Daucus spp.

                  Foeniculum vulgare
                  Mill.

                  Mulinum spinosum *    62   52   87   61

Asteraceae        Anthemideae           m    m

                  Astereae *            m    m         m

                  Cirsium vulgare            m
                  (Savi) Ten

                  Cichorium intybus     m    m         m
                  L.

                  Cichorieae                           m

                  Hypochoeris
                  radicata L.

                  Leucheria                  m
                  achillaeifolia *

                  Taraxacum             m    m
                  officinale
                  F. H. Wigg.

                  Senecio spp. *        m    m    m    m

Betulaceae        Betula spp.

Boraginaceae      Phacelia                   m
                  secunda J.F.
                  Gmel. *

Brassicaceae      Brassicaceae          M    m    m    M

Caprifoliaceae    Lonicera spp.

Caryophyllaceae   Caryophyllaceae

Chenopodiaceae    Chenopodiaceae (a)    m

Convolvulaceae    Convolvulus           m         m
                  arvensis L.

Cyperaceae        Cyperaceae (a)

Ephedraceae       Ephedra spp. *

Fabaceae          Fabaceae

                  Adesmia spp. *        m    m

                  Lotus spp.

                  Medicago sativa L.         m         m

                  Melilotus sp.         M    M    M    S

                  Trifolium spp.        m    m         m

                  Vicia spp.            m    m    m

Geraniaceae       Erodium
                  cicutarium
                  (L.) Aiton

Juglandaceae      Juglans spp. (a)

Malvaceae         Malva spp.

Myrtaceae         Eucalyptus spp.

Oleaceae          Ligustrum spp.        m

Onagraceae        Oenothera spp. *

Pinaceae          Pinus sp. (a)

Plantaginaceae    Plantago sp. (a)           m    m    m

Poaceae           Poaceae (a)                     m

Polygalaceae      Polygala spp. *

Polygonaceae      Rumex acetosella
                  L. *

Rhamnaceae        OchetophiIa           m              m
                  trinervis
                  (Gillies ex Hook
                  & Arn.) Poepp. *

Rosaceae          Rosaceae                   S

                  Acaena spp. *         m    m    m

Schoepfiaceae     Quinchamalium
                  chilense
                  Molina *

Typhaceae         Typha sp. (a)         m

Verbenaceae       Mulguraea-
                  Junellia spp. *

Family                                      Samples

                                        5    6    7    8

Anacardiaceae     Schinus marchandii    m    m
                  F. A. Barkley *

Apiaceae          Conium maculatum L.   m              m

                  Daucus spp.                     m    m

                  Foeniculum vulgare
                  Mill.

                  Mulinum spinosum *    61   64   63   64

Asteraceae        Anthemideae           m

                  Astereae *            m    m    m

                  Cirsium vulgare
                  (Savi) Ten

                  Cichorium intybus
                  L.

                  Cichorieae

                  Hypochoeris
                  radicata L.

                  Leucheria                       m    m
                  achillaeifolia *

                  Taraxacum             m    m    m    m
                  officinale
                  F. H. Wigg.

                  Senecio spp. *        m    m    M    m

Betulaceae        Betula spp.

Boraginaceae      Phacelia                        m
                  secunda J.F.
                  Gmel. *

Brassicaceae      Brassicaceae          m    M    M    M

Caprifoliaceae    Lonicera spp.

Caryophyllaceae   Caryophyllaceae

Chenopodiaceae    Chenopodiaceae (a)         m

Convolvulaceae    Convolvulus           m    m    m    m
                  arvensis L.

Cyperaceae        Cyperaceae (a)

Ephedraceae       Ephedra spp. *

Fabaceae          Fabaceae

                  Adesmia spp. *

                  Lotus spp.                           m

                  Medicago sativa L.    m    m    m    m

                  Melilotus sp.         S    S    M    S

                  Trifolium spp.        M    M         m

                  Vicia spp.

Geraniaceae       Erodium
                  cicutarium
                  (L.) Aiton

Juglandaceae      Juglans spp. (a)

Malvaceae         Malva spp.            m    m         m

Myrtaceae         Eucalyptus spp.            m

Oleaceae          Ligustrum spp.

Onagraceae        Oenothera spp. *

Pinaceae          Pinus sp. (a)

Plantaginaceae    Plantago sp. (a)      M    m    m    m

Poaceae           Poaceae (a)           m    m         m

Polygalaceae      Polygala spp. *

Polygonaceae      Rumex acetosella                m
                  L. *

Rhamnaceae        OchetophiIa                m    m
                  trinervis
                  (Gillies ex Hook
                  & Arn.) Poepp. *

Rosaceae          Rosaceae              m         m    m

                  Acaena spp. *         m    m    m    m

Schoepfiaceae     Quinchamalium
                  chilense
                  Molina *

Typhaceae         Typha sp. (a)

Verbenaceae       Mulguraea-            m
                  Junellia spp. *

Family                                       Samples

                                        9    10   11   12

Anacardiaceae     Schinus marchandii
                  F. A. Barkley *

Apiaceae          Conium maculatum L.

                  Daucus spp.

                  Foeniculum vulgare
                  Mill.

                  Mulinum spinosum *    65   58   60   69

Asteraceae        Anthemideae                     m

                  Astereae *                 m    m    m

                  Cirsium vulgare                 m
                  (Savi) Ten

                  Cichorium intybus     m         m
                  L.

                  Cichorieae

                  Hypochoeris                m
                  radicata L.

                  Leucheria             m
                  achillaeifolia *

                  Taraxacum             m    m    m    m
                  officinale
                  F. H. Wigg.

                  Senecio spp. *        m    m    m    m

Betulaceae        Betula spp.                          m

Boraginaceae      Phacelia              m    m    m
                  secunda J.F.
                  Gmel. *

Brassicaceae      Brassicaceae          M    M    M    M

Caprifoliaceae    Lonicera spp.

Caryophyllaceae   Caryophyllaceae

Chenopodiaceae    Chenopodiaceae (a)

Convolvulaceae    Convolvulus           m    m    m    m
                  arvensis L.

Cyperaceae        Cyperaceae (a)                       m

Ephedraceae       Ephedra spp. *

Fabaceae          Fabaceae

                  Adesmia spp. *        m              M

                  Lotus spp.            m

                  Medicago sativa L.    m    M    m

                  Melilotus sp.         S    S    S    M

                  Trifolium spp.             m    m    m

                  Vicia spp.

Geraniaceae       Erodium                              m
                  cicutarium
                  (L.) Aiton

Juglandaceae      Juglans spp. (a)

Malvaceae         Malva spp.                 m    m

Myrtaceae         Eucalyptus spp.                      m

Oleaceae          Ligustrum spp.

Onagraceae        Oenothera spp. *

Pinaceae          Pinus sp. (a)

Plantaginaceae    Plantago sp. (a)      m    m    M    m

Poaceae           Poaceae (a)                     m    m

Polygalaceae      Polygala spp. *            m

Polygonaceae      Rumex acetosella
                  L. *

Rhamnaceae        OchetophiIa           m    m
                  trinervis
                  (Gillies ex Hook
                  & Arn.) Poepp. *

Rosaceae          Rosaceae                        m    m

                  Acaena spp. *              m    m    m

Schoepfiaceae     Quinchamalium
                  chilense
                  Molina *

Typhaceae         Typha sp. (a)                   m

Verbenaceae       Mulguraea-                 m
                  Junellia spp. *

Family                                       Samples

                                        13   14   15   16

Anacardiaceae     Schinus marchandii
                  F. A. Barkley *

Apiaceae          Conium maculatum L.

                  Daucus spp.                m

                  Foeniculum vulgare    m
                  Mill.

                  Mulinum spinosum *    55   76   62   73

Asteraceae        Anthemideae

                  Astereae *            m    m         m

                  Cirsium vulgare       m              m
                  (Savi) Ten

                  Cichorium intybus
                  L.

                  Cichorieae

                  Hypochoeris
                  radicata L.

                  Leucheria
                  achillaeifolia *

                  Taraxacum                       m
                  officinale
                  F. H. Wigg.

                  Senecio spp. *        m    m    m    m

Betulaceae        Betula spp.

Boraginaceae      Phacelia                             m
                  secunda J.F.
                  Gmel. *

Brassicaceae      Brassicaceae          M    M    M    S

Caprifoliaceae    Lonicera spp.         m

Caryophyllaceae   Caryophyllaceae       m

Chenopodiaceae    Chenopodiaceae (a)

Convolvulaceae    Convolvulus           m    m         m
                  arvensis L.

Cyperaceae        Cyperaceae (a)                       m

Ephedraceae       Ephedra spp. *             m

Fabaceae          Fabaceae                   m    m

                  Adesmia spp. *             m    m

                  Lotus spp.                 m         m

                  Medicago sativa L.    M    m    m

                  Melilotus sp.         S    M    M    M

                  Trifolium spp.        m    m    m

                  Vicia spp.                 m

Geraniaceae       Erodium
                  cicutarium
                  (L.) Aiton

Juglandaceae      Juglans spp. (a)           m

Malvaceae         Malva spp.            m

Myrtaceae         Eucalyptus spp.

Oleaceae          Ligustrum spp.

Onagraceae        Oenothera spp. *      m

Pinaceae          Pinus sp. (a)                        m

Plantaginaceae    Plantago sp. (a)      M    m    m    m

Poaceae           Poaceae (a)           m              m

Polygalaceae      Polygala spp. *                 m

Polygonaceae      Rumex acetosella      m    m
                  L. *

Rhamnaceae        OchetophiIa
                  trinervis
                  (Gillies ex Hook
                  & Arn.) Poepp. *

Rosaceae          Rosaceae              m              m

                  Acaena spp. *         m    M    S    m

Schoepfiaceae     Quinchamalium                   m
                  chilense
                  Molina *

Typhaceae         Typha sp. (a)

Verbenaceae       Mulguraea-                      m
                  Junellia spp. *

For the predominant (>45%) pollen (Mulinum spinosum) the numbers
refer to found percentages. *: native plants; (a): Pollen from
wind-pollinated plants; S: secondary pollen (16-45%); M: important
minor pollen (3-15%); m: minor pollen (<3%).

TABLE II

RESULTS OF THE QUANTITATIVE POLLEN ANALYSIS

Samples     NPG       Group      HDE/P
                    (Maurizio)

1          22,220       II       0.01
2          35,111       II       0.01
3         110,666      III       0.00
4         112,838      III       0.00
5         101,443      III       0.00
6         136,000      III       0.00
7          44,959       II       0.00
8          32,621       II       0.01
9          58,750       II       0.01
10        117,030      III       0.01
11         30,973       II       0.01
12         42,491       II       0.00
13         40,452       II       0.00
14        141,425      III       0.00
15         51,325       II       0.81
16         71,628       II       0.01

NPG: Number of pollen grains in 10g of honey;
HDE/P: Honeydew index (ratio of honeydew
elements to pollen grains of nectariferous
plants) Group according to Maurizio (1939).

TABLE III

PHYSICO-CHEMICAL PARAMETERS OF Mulinum spinosum HONEYS

Sample   Moisture     Color       pH        Free          Elect.
           (%)      (mm Pfund)            acidity         cond.
                                           (meq x         (mS x
                                        [kg.sup.-1)]   [cm.sup.-1)]

1          11.4        61.3      3.7         18            0.29
2          17.1        94.3      4.3         21            0.37
3          14.9        74.1      4.6         17            0.41
4          16.2        79.8      4.6         24            0.34
5          15.5        52.5      4.4         25            0.33
6          15.6        80.4      4.5         20            0.32
7          16.6         30       4.5         19            0.43
8          15.1        71.5      4.5         31            0.5
9          15.3        71.5      4.7         17            0.45
10         14.3        97.1       5          22            0.65
11         14.7         40       5.3         19            0.41
12         16.1         45       4.1         15            0.34
13         14.5         40       4.2         19            0.57
14         14.4         25       4.1         19            0.39
15         14.6         35       4.1         20            0.44
16           0          45       4.1         17            0.34
Mean      15.19        58.9      4.42       20.2           0.41
Max        17.1        97.1      5.3         31            0.65
Min        11.4         25       3.7         15            0.29
SD         1.35        22.9      0.39        0.1           0.07

Sample    Ashes    HMF-1 (mg x    Diastase    Polyphenols
         (% w/w)   [kg.sup.-1)]     (DN)     (mg GAE/100 g)

1         0.086        0.15         16.8          190
2         0.132        4.19         18.6          130
3         0.155        1.65         17.5          158
4         0.15         3.44         14.1          134
5         0.11         4.34          9.9          130
6          0.1         4.19          5.9          133
7         0.17         4.49          10           146
8           0          4.34          7.1          130
9         0.178        3.76          7.4          130
10        0.293        3.64          9.8          163
11        0.155        2.25          5.6          141
12        0.115        3.74          5.3          169
13        0.247        3.52         30.5          113
14        0.144         0           14.5          152
15        0.172        1.2          14.5          119
16        0.115         0           14.4          151
Mean      0.15         2.81         12.6         143.1
Max       0.293        4.49         30.5          190
Min         0           0            5.3          113
SD        1.67         6.52          20            1.7

TABLE IV

COMPOSITION AND TOTAL AMOUNT (g/100g) OF MONO-DI AND
OLIGOSACCHARIDES IN Mulinum spinosum HONEYS

Samples *   Fructose   Glucose   Turanose   Trehalose

2             39.4      26.3       2.9         1.1
3             40.3      27.0       2.1         0.7
4             40.0      24.2       2.4         0.9
5             37.7      31.4       2.2         0.8
6             38.3      28.0       2.3         0.7
7             40.3      28.2       2.1         0.6
8             37.9      29.8       2.0         0.7
9             37.8      29.0       2.1         0.9
10            38.0       8.8       2.1         1.0
11            40.4      29.1       2.1         1.0
12            40.0      25.8       2.4         0.7
13            36.3      31.3       2.4         1.4
14            39.4      34.0       1.8         0.6
15            38.3      32.3       1.8         0.7
16            37.8      37.4       1.4         0.6

Samples *   Erlose   Melezitose   Raffinose   Maltotriose

2                       0.6
3
4
5                                    0.1
6
7
8
9            0.2        0.2
10                      0.7                       0.6
11           0.4
12                      0.2
13           0.4        0.5                       0.3
14           0.1
15                      0.7
16

* Sample 1 was not considered for this analysis because
the amount of honey insufficient to perform the assays.
Saccharose, maltose and maltotetraose were not detected.
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Title Annotation:articulo en ingles
Author:Aloisi, Pia V.; Forcone, Alicia E.; Amadei, Marisa
Publication:Interciencia
Date:Jul 1, 2013
Words:5496
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