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Physicochemical characteristics and pollen spectrum of some Algerian honeys


Salim Ouchemoukh a, Hayette Louaileche
a

b,*

, Paul Schweitzer

partement de Biologie Physico-chimique, Faculte des Sciences de la Nature et de la Vie, Universite de Bejaia, De Route de Targa-Ouzemour 06000 Bejaia, Algeria b partement des Sciences Alimentaires, Faculte des Sciences de la Nature et de la Vie, Universite de Bejaia, De Route de Targa-Ouzemour 06000 Bejaia, Algeria nange, France CETAM Lorraine, Laboratoire dAnalyses et dEcologie Apicole, 1B, Rue Jeanne dArc 57310 Gue Received 1 September 2004; received in revised form 4 August 2005; accepted 7 August 2005

Abstract The characterisation of Algerian honeys was carried out on the basis of the microscopic (pollen analysis) and physico-chemical properties. The samples were analysed for parameters including refractive index, moisture, density, dynamic viscosity, pH, specic rotation, electrical conductivity, ash, sugars, proteins, proline and phenolic compounds contents. The results obtained in the present study show the variability of chemical composition of the honey samples. The botanical families Myrtaceae, Apiaceae and Ericaceae are most frequently found. The most pollen types are Eucalyptus which is present in ve samples. The samples are found to meet national and international honey specications. 2005 Elsevier Ltd. All rights reserved.
Keywords: Honey; Physico-chemical characterisation; Botanical origin; Pollen analysis

1. Introduction Honey is produced by honeybees from nectar of plants, as well as from honeydew. This latter is a sugar-containing substance excreted by some plant-sucking insects. Honey contains at least 181 substances (Louveaux, 1985; Sato & Miyata, 2000). Chemically, honey comprises sugars (70 80%), water (1020%) and other minor constituents such as organic acids, mineral salts, vitamins, proteins, phenolic compounds and free amino acids. The monosaccharides, fructose and glucose, are the main sugars found in honey (Jean-Prost, 1987; Nagai, Inoue, Inoue, & Suzuki, 2002; rez, Diez, & Heredia, 2001). Amino acids Terrab, Vega-Pe account for 1% and proline is the major contributor with 5080% of the total amino acids (Hermosin, Chicon, & Dolores Cabezudo, 2003).

Corresponding author. E-mail address: haylouaileche@yahoo.fr (H. Louaileche).

Studies have shown that honey has both antimicrobial and antioxidant properties, useful in stimulation of wounds and burns healing and gastric ulcers treatment (Gheldof & Engeseth, 2002). The antimicrobial properties of honey is well documented (Al-Somal, Coley, Molan, & Hancock, 1994; Brady, Molan, & Harfoot, 1996; Molan, 1992). Physical and chemical properties of dierent kinds of honey have been reported by many scientists (RodriguezOtero, Paeiro, Simal, Terradillos, & Cepeda, 1992; Cano, Felsner, Bruns, Whatanabe, & Almeida-Muradian, 2001; Persano Oddo, Piazza, Sabatini, & Accorti, 1995; Yilmaz & Yavuz, 1999). Pollen analysis has been the traditional method to determine the oral origin of the honey, but this technique is tedious and has some limitations (Hermosin et al., 2003; Von der Ohe, 1994). Usually, honey is considered unioral, if the pollen frequency of that plant is >45%. Some pollen grains, such as Citrus and members of the Lamiaceae family, are under-represented in the honey spectra, while others (Eucalyptus, Castanea and Myosotis) are over-represented

0956-7135/$ - see front matter 2005 Elsevier Ltd. All rights reserved. doi:10.1016/j.foodcont.2005.08.007

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(Terrab, Diez, & Heredia, 2003c). A minimum of 10% Citrus sp. pollen is enough to consider a honey as unioral (Terrab, Diez, & Heredia, 2003b). Mateo and Bosch-Reig (1998) suggested the use of physico-chemical criteria such as electrical conductivity and pH analyses complemented by pollen analysis for characterisation of unioral honeys. Electrical conductivity, specic rotation, ash content and pH are widely used for discrimination between honeydew and blossom honeys. Electrical conductivity is a good criterion of the botanical origin of honey. Blossom honeys and mixtures of blossom and honeydew honeys should have less than 0.8 mS/cm and honeydew and chestnut honeys should have more than 0.8 mS/cm. The measurement of specic rotation is currently used in Greece, Italy and UK to distinguish between blossom and honeydew honeys. Most of the honeydew honeys have positive values of specic rotation whereas nectar honeys have negative one. This is a consequence of the normal preponderance of fructose in oral honey, which shows a negative specic rotation over glucose (Bogdanov et al., 1999). Beja a is among the important honey producer in Algeria, since it is suitable for apiculture. In 2003, the local production is estimated at 85 tons. Therefore, the present study was undertaken to characterise the physico-chemical properties and the botanical origin (blossom and/or honeydew honeys) of Beja a samples honey.

2. Materials and methods 2.1. Honey samples Eleven samples of honeys produced in various regions of Beja a (Algeria) (Fig. 1) were collected from beekeepers in 2002. The samples were stored in a refrigerator in airtight plastic containers until analysis. The regions from which the samples of honey were collected are indicated in Table 1. Analyses were carried out at least in duplicate. 2.2. Pollen analysis Pollen analysis was carried out using the methods established by the International Commission of Bee Botany described by Louveaux, Maurizio, and Vorwohl (1978). 2.3. Physico-chemical parameters Water content (moisture) was determined by refractometry (Journal Ociel Franc ais, 1977) using Abbe-typ refractometer (RF 490, Euromexholland). Density and dynamic viscosity were determined according to Bogdanov et al. (1995). Density gravity was determined by dividing the weight of specic gravity bottle (10 ml) lled with honey to the weight of the same bottle,

Fig. 1. Distribution of the samples in the Beja a region (northeast Algeria).

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Table 1 Samples of Beja a honeys and their botanical origin Samples H01 H02 H03 H04 H05 H06 H07 H08 H09 H10 H11 Location Oued-Des Tizi-Nberber Kherrata Tala-Hamza Aokas Oued-Ghir Amizour El-kseur Souk-El-Tenine Amizour Ihaddaden Botanical origin Honeydew Blossom/honeydew Blossom (monooral) Blossom (monooral) Blossom (monooral) Blossom (monooral) Blossom (monooral) Blossom (polyoral) Blossom (polyoral) Blossom (monooral) Blossom (polyoral)

lled with distilled water. Dynamic viscosity was determined at 30 C using FENSK viscosimeter. The pH was assessed in a 10% (w/v) solution of honey in distilled water (Journal Ociel Franc ais, 1977) by mean of pH meter (CRISON 2000). Ash and proline contents, electrical conductivity, and specic rotation were determined by the methods of Bogdanov et al. (1997). Ash content was determined by heating 5 g of honey at 625 C in a mue furnace. Proline content was determined by the measurement of the absorbance at 510 nm of the resulting product between proline and ninhydrin in an acidic medium. Electrical conductivity was measured in a 20% (w/v) solution of honey in deionized water with low electrical conductivity (<14 lS/cm) using Leibohld model conductimeter. Specic rotation was measured in a polarimeter as follows: 12 g of honey sample was claried by Carrez reagents (I and II), and distilled water was added to get a nal volume of 100 ml. Then, the obtained solution was inserted into the polarimeter. Total reducing sugars, reducing and non-reducing sugars were determined by the methods of Journal Ociel , Figarella, and Zonszain (1984) Franc ais (1977), Audigie and Salgarolo (1990). Aldoses content was determined by the method of Gonnet (1986). The concentration of total phenolic compounds were determined by the Folin-Ciocalteu reagent (Al-Mamary, Al-Meeri, & Al-Habori, 2002) and expressed in mg/100 g of honey as gallic acid equivalent. The protein content was determined by the method of Azeredo, Azeredo, De Souza, and Dutra (2003). A volume of 0.1 ml of protein extract (honey sample 50% w/v) was added to 5 ml of Coomassie Brillant Blue. After 2 min of incubation, the quantity of proteins was estimated at 595 nm in relation to bovine serum albumin standard curve. 3. Results and discussion 3.1. Pollen analysis The results from the pollen analysis, summarised in Table 2, show that predominant pollen is found in 8 sam-

ples and honey samples of El-Kseur (H08), Souk-El-Tenine (H09) and Ihaddaden (H11) are polyoral honeys. More than 13 pollen types are found in honey samples of Tizi-Nberber (H02), Kherrata (H03) and Souk-El-Tenine (H09), collected in the eastern part of Beja a, 11 13 in samples of Oued-Ghir (H06), Amizour (H10) and H11, and 610 in others. The samples are low in sediments: the number of pollen grains in 10 g of honey ranged between 20,000 and 40,000. The frequency classes of pollen grains are given as predominant pollen (>45%), secondary pollen (1645%), important minor pollen (315%) and minor pollen (13%). All the samples fell into class II of Maurizio. Pollen types of honey samples correspond to more seventeen families. The number of botanical elements is not determined but samples H01 and H02 show the presence of a lot of honeydew elements: fungal spores and mycelium. Also, these samples have pollen grains from anemophilus species (Olea, Pinaceae, Quercus, Cistus). Blossom (oral) honeys contain pollen from entomophilus species, while the pollen in the honeydew honeys, is from s, & Terrab, 2004). Myranemophilus species (Diez, Andre tus communis pollen is present at 79% in Oued-Des honey (H01), it is a honeydew honey. Tizi-Nberber honey (H02) contains the predominant pollen of Rosaceae family, Rubus (82%). This honey is a mixture of blossom and honeydew honey. Ericaceae are important melliferous plants in the Mediterranean area (Terrab et al., 2003c). The pollen of Erica arborea is predominant in Oued-Ghir honey (H06) and it is secondary pollen in Souk-El-Tenine honey (H09). It is generally accepted that a minimum content of 70% Eucalyptus pollen is necessary to classify an Eucalyptus honey as unioral. Eucalyptus honeys are considered among the best honeys and are very valuable from a consumers point of view (Terrab, Diez, & Heredia, 2003a). Therefore, honeys harvested in Amizour are monooral honeys Eucalyptus. This latter and Myrtus communis belong to Myrtaceae family. The pollen grains of Aokas honey (H05) are overrepresented. It is dicult for us to explain the pollen spectra of the Ihaddaden honey (H11); the majority of the pollen types of this honey are from tropical plants area (Mimosa pudica: 41%, Mimosa bimucromata: 6% and Myrtaceae: 24%). 3.2. Physico-chemical parameters The results of physico-chemical analyses of honey from dierent sources are summarised in Tables 3 and 4. Water content, a parameter related to the maturity degree, is between 14.64% and 19.04%, corresponding to refractive index between 1.4889 and 1.4999. Oued-Des honey (H01) has the highest moisture content as compared to other samples. There is a variation in water content among the investigated honeys. Nanda, Sarkar, Sharma, and Bawa (2003) stated that moisture content is aected by climate, season and moisture content of original plant. The values obtained are below 20%, and the maximum allowed by the European Community Directive (The Council of the European Un-

S. Ouchemoukh et al. / Food Control 18 (2007) 5258 Table 2 Pollen types present in the honey samples (in percentages) Samples H01 H02 Predominant pollen (>45%) Myrtus communis 79 Rubus 82 Secondary pollen (1645%) Minor pollen (315%) Erica arborea 14 Inula spp 8, Erica arborea 4 Important minor pollen (<3%)

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H03

Capparis spp 50

Prunus/Pyrus 19, Rubus 17

Apiace 6

H04 H05 H06

Hedysarum coronarium 53 Eucalyptus 63 Erica arborea 54

Eucalyptus spp 42 Hedysarum coronarium 9, Rubus 7, Apiace 4 Rubus 7

Lavandula stchas, Carduus, Prunus/Pyrus, Lamiace, Asterace type matricaria,. . . Hedysarium coronarium 2, Apiace, Lavandula stchas, type solidago, Asterace liguliore, Liliace, Malvace, Asphodelus spp, Scabiosa spp, Euphorbia spp, Prunus/Pyrus, Hmatoxylon campechianum Carduus, Brassicace, Inula spp, Asterace liguliore, Hedysarium coronarium, Eucalyptus spp, Asphodelus, Prunus dulcis, Allium spp, Trifolium spp, Echinops spp, Dipsacace, thymus Rubus, Carduus, Fabace, Trifolium spp, Acacia spp, Dipsacace, Convolvulus Carduus, Citrus Carduus, Brassicace, citrus, Lavandula stchas, Lamiace, Hedysarum coronarium, Acacia spp, Apiace, Convolvulus Malvace, Convolvulus, Acacia spp Asterace, Inula spp, Carduus, Apiace, Acacia spp

Eucalyptus spp 27

H07 H08

Eucalyptus 87 Eucalyptus spp 44, Erica arborea 28 Erica arborea 33

H09

H10

Eucalyptus 85

Brassicace 7, Citrus 6 Citrus 11, Hedysarum coronarium 5, Lavandula stchas 4 Punica granatum 12, Hedysarum coronarium 9, Eucalyptus 7, Brassicace 7, Trifolium spp 4, type Genista 4 Citrus 5

Lavandula stchas, Apiace, Lamiace, Apiace type buplevrum, Carduus, Prunus dulcis, Asterace liguliore, Acacia spp,. . . Acacia spp, type buplevrum, Prunus/Pyrus, Rubus, ranunculace, Lamiace, Centaurea, Anthyllis, Vicia, Inula, Convolvulus. . . Type leptospermum, Trifolium spp, Carduus, Acacia spp, Anacardiace (mangifera spp)

H11

Mimosa pudica 41, Myrtace 24

Echium 10, Asterace type Bidens10, type Mimosa bimucromata 6, Hmatoxylon campachianum 3

Table 3 Some chemical characteristics of honey samples Samples Moisture (%) 19.04 0.56 17.04 0.00 14.64 0.28 18.44 0.28 18.64 0.56 17.88 0.00 16.80 0.22 16.54 0.14 16.64 0.28 15.84 0.00 16.84 0.42 pH Ash (%) Electrical Aldoses conductivity (%) (mS/cm) 1.61 0.09 0.60 0.03 0.21 0.01 0.54 0.04 0.65 0.05 0.61 0.05 0.41 0.03 0.47 0.03 0.51 0.03 0.33 0.02 0.23 0.02 36.90 0.00 35.10 0.73 34.80 0.85 37.50 0.42 39.00 0.42 42.00 0.42 40.50 0.73 39.90 0.73 39.30 1.12 40.80 0.42 28.50 0.42 Total reducing sugars (%) 73.00 2.12 71.25 4.59 77.00 0.70 73.00 2.12 81.00 0.00 75.50 7.77 79.50 4.24 77.50 0.00 77.00 0.70 84.25 6.71 80.75 4.59 Reducing sugars (%) 67.83 3.06 70.25 1.76 73.25 4.59 68.58 4.12 72.25 1.06 75.41 2.94 73.91 5.06 77.25 1.06 73.41 1.53 80.25 1.06 77.83 4.48 Sucrose Proteins (%) (mg/g) 4.91 0.95 3.56 4.19 8.31 0.08 5.31 0.23 3.41 3.80 2.77 8.6 0.3 6.3 0.4 3.8 0.6 6.0 0.1 6.6 0.4 9.4 0.1 5.5 0.3 6.9 0.1 5.6 0.3 3.7 0.2 3.7 0.0 Proline (mg/kg) Phenolic compounds (mg/100 g)

H01 H02 H03 H04 H05 H06 H07 H08 H09 H10 H11

4.43 0.01 3.98 0.05 3.79 0.04 3.49 0.01 3.50 0.04 3.69 0.03 3.64 0.01 3.60 0.02 3.87 0.05 3.56 0.03 3.76 0.04

0.54 0.00 0.23 0.01 0.06 0.01 0.15 0.01 0.09 0.00 0.19 0.041 0.20 0.00 0.18 0.04 0.21 0.01 0.16 0.02 0.09 0.00

680 17 1304 5 346 11 714 10 212 08 64 10 393 36 157 30 434 15 461 15 425 06 657 20 378 23 132 5 481 25 468 15 379 06 307 10 421 21 79 0 202 13 271 10

ion, 2002). The quality of the studied honey is good because honey with high water content is more likely to ferment (Bogdanov et al., 1999). Water content is highly important for the shelf-life of the honey during storage (Terrab et al., 2003b). The physical parameters density and dynamic viscosity values ranged from 1.4009 to 1.4505 and from 1.12 to

8.98 mPa s, respectively. Kherrata honey (H03) shows the highest density and viscosity. Generally, the honey samples having a high moisture content had the lowest density and vice versa. All honeys are acidic, having a pH in the range 3.49 4.43. The acidity of honey is due to the presence of organic acids, particularly the gluconic acid and inorganic ions

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Table 4 Some physical characteristics of Beja a honey samples Samples H01 H02 H03 H04 H05 H06 H07 H08 H09 H10 H11 Refractive index 1.4889 0.0141 1.4939 0.0000 1.4999 0.0008 1.4904 0.0007 1.4899 0.0014 1.4918 0.0000 1.4945 0.0005 1.4951 0.0003 1.4949 0.0007 1.4969 0.0000 1.4936 0.0010 Specic gravity 1.4073 0.0075 1.4106 0.0079 1.4505 0.0086 1.4068 0.0063 1.4009 0.0054 1.4107 0.0070 1.4152 0.0147 1.4191 0.0053 1.4197 0.0059 1.4095 0.0111 1.4165 0.0026 Dynamic viscosity (mPa s) 3.57 0.02 4.16 0.03 8.98 0.12 1.61 0.01 1.12 0.01 3.00 0.20 2.76 0.00 3.65 0.02 3.94 0.02 3.10 0.01 2.24 0.02 Specic rotation +2.83 0.06 2.33 0.43 +1.85 0.04 5.80 0.06 1.99 1.18 5.72 0.25 6.20 0.46 5.65 1.18 1.00 0.03 6.60 0.28 7.29 0.14

such as phosphate and chloride (Nanda et al., 2003). These results agreed with data reported by Azeredo et al. (2003). The pH of honeydew and blossom honey blends is between 3.5 and 4.5. The pH of honeydew honey is between 4.5 and 5.5 (Gonnet, 1986). Those of Switzerland, Moroccan honeydew honey, Eucalyptus and multioral honeys are 4.4 (Bogdanov, 1997), 4.28, 3.65 and 3.72 (Terrab, Diez, & Heredia, 2002), respectively. The ash content of the studied honey samples diers widely. It ranged from 0.06% to 0.54%. The maximum value is found for the sample H01, followed by the sample H02 (0.23%). These dierences in mineral content are dependent on the type of soil in which the original nectar bearing plant was located (Anklam, 1998). The ash content is a quality criterion for honey botanical origin; the blossom honeys have lower ash content than honeydew honeys. The mineral content of blossom honey is 60.6%; the value of honeydew or blends of honeydew and blossom honey or chestnut honey is 61.2%. This parameter is generally replaced by the measurement of electrical conductivity (Bogdanov et al., 1999). Popek (2002) demonstrated that the ash content honeydew honey is 0.56%. Honeydew and/or mixed honeys have the highest ash content (Diez et al., 2004). The electrical conductivity of ten honeys analysed is less than 0.70 mS/cm. Oued-Des honey has the highest conductivity (1.61 mS/cm). This parameter depends on the ash, organic acids, proteins, some complex sugars and polyols content, and varies with botanical origin (Terrab et al., 2003a). The conductivity measurement is easy, fast and needs only inexpensive instrumentation. It is widely used for discrimination between honeydew and blossom honeys and also for the characterisation of unioral honeys (Bogdanov et al., 1999). The honeydew honeys are characterised by their very dark colour and high values of pH, ash content and electrical conductivity (Thrasyvoulou & Manikis, 1995; Diez et al., 2004). Therefore, Oued-Des honey (H01) is a honeydew honey. In the present study, the coecient of correlation between electrical conductivity and ash content is 0.92. Specic rotation is levorotatory in most honey samples. Only two honey samples (H01 and H03) are dextrorota-

tory. The specic rotation of the Oued-Des sample conrms that this honey is a honeydew honey. Although Kherrata honey has positive specic rotation, pollen analysis, ash content, pH and electrical conductivity indicate that this honey is a blossom honey. Data about pH, ash content, electrical conductivity and specic rotation indicate that the samples H01 (Oued-Des) and H02 (Tizi-Nberber) are honeydew honey and mixed honey, respectively; others are blossom (oral) honeys. The major sugars present in honey are fructose and glucose. The amounts of total reducing sugars and reducing sugars vary from 71.25% to 84.25% and from 67.83% to 80.25%, respectively. These results conrm that sugars represent the major constituents of honey. Honeydew honeys contain less monosaccharides and more di, tri and higher oligosaccharides than blossom (nectar) honeys (Diez et al., 2004). Honeydew honeys have the highest content of melezitose (Mateo & Bosch-Reig, 1998). Sample H01 has a low amount of reducing sugars (67.80%), it is a honeydew honey. Glucose is the main aldose in the honey. Aldoses values ranged from 29.4% to 42.0%. Ihaddaden honey has the lowest aldoses content compared to others. Therefore, it has more fructose than other samples. This result is conrmed by specic rotation, Ihaddaden honey is more levorotatory. Sucrose (saccharose) contents ranged from 0.08% to 5.31%. Only the honey sample of Aokas (H05) has 8.31% of sucrose, an important sugar from the legislative point of view. The limit of sucrose content for Eucalyptus honey allowed by the European Community Directive (The Council of the European Union, 2002) is 610%. The majority of the pollen types of sample H05 are Eucalyptus (44%). Studied honeys are authentic, because the obtained results complied with requirements of the Algerian Quality Standards (Anonymous, 1995). The sucrose level can be increased if the beekeeper has over-fed the bees with sugar during the spring (Anklam, 1998). Moreover, a high content of this sugar means an early harvest of the honey (Azeredo et al., 2003). The sucrose contents for the samples H02, H06 and H08 are 0.95%, 0.08% and 0.23%, respectively. These values could be attributed to the action of

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invertase because the amount of saccharose can be decrease during honey storage due to the presence of the enzyme (Anklam, 1998). Proline comes mainly from the salivate secretions of Apis mellifera during the conversion of nectar into honey (Bergner & Hahn, 1972). The concentrations of proline ranged between 202 and 680 mg/kg. Some authors report that high values of proline are typical for honeydew honeys. Proline content is a criterion of honey ripeness and in some cases, also of sugar adulteration. A minimum value for guenine honey of 180 mg/kg honey is accepted in honey control laboratories (Bogdanov et al., 1999). Consequently, studied honeys are ripened and not adulterated. Amino acids (proline, histidine, glycine, alanine) are one of the antioxidant components in honey. The correlation between radical scavenging activity (RSA) and proline content is higher than between RSA and total phenolic content (Meda, Lamien, Romito, Millogo, & Nacoulma, 2005). The protein content of honey is normally less than 5 mg/ g (Anklam, 1998). The protein contents of analysed honey samples were between 3.7 and 9.4 mg/g. The results of this study are higher than those obtained by Azeredo et al. (2003) for honeys of Borreria verticillata (2.23 mg/g), known in Brazil as vassourinha and recommended as alimentary complements for the population of the northeast area of the country. Honeys contain a great number of phenolic compounds, the nature and the quantity of which vary widely according to the oral origin (Amiot, Aubert, Gonnet, & Tacchini, 1989). The total phenolic compounds ranged from 64 mg/ 100 g for Kherrata honey to 1304 mg/100 for Oued-Des honey. Many researchers found that honeys with dark color has a higher amount of total phenolic compounds. Gheldof and Engeseth (2002), Meda et al. (2005) have demonstrated a correlation between antioxidant activity and the total phenolic content. Meda et al. (2005) found that honeydew honey has the highest amount of total phenolic compounds (114.75 mg/100 g). According to Al-Mamary et al. (2002), the determination of the total phenolic content of honey is a good parameter for the assessment of its quality and possible therapeutic potential. Oued-Des honey would present the best antioxidant and antimicrobial activities. Investigations reveal that the analysis of avono ds and other phenolic compounds constitute a very promising technique to study the oral origin of honey. For instance, sunower honeys contained an important relative amount of quercetin (Soler, Gil, Garcia-Viguera, & Tomas-Barberan, 1995). 4. Conclusion The present study describes the variability of some physicochemical characteristics of eleven Algerian honeys. The results obtained agreed with requirements of European Community Directive. They also demonstrate, that botanical origin (blossom and/or honeydew honeys) can be determined by physico-chemical parameters. Studied sam-

ples are found to be low in moisture and therefore safe from fermentation. Oued-Des honey shows the best honeydew honey characteristics with high pH, ash content, electrical conductivity and low reducing sugars content. The large variety of melliferous sources enables Algeria to produce characteristic honeys. For instance, predominant pollen in Kherrata honey is Capparis spp and no previous studies for this type are reported. Extensive research is required to establish therapeutic (antioxidant and antimicrobial) properties of Algerian honeys. Acknowledgements We are grateful to beekeepers that collaborated with us in providing honey samples. We thank Miss Razika Louaileche for kindly correcting the English version of the manuscript and the referees for their comments. References
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