Honey and Its Products. Chemical, Biological and Therapeutic Applications

Table of Contents

I. Introduction

II – REVIEW OF LITERATURE
2.1. Honeybee activity in collecting and producing some bee products:
2.2. Biological properties of some bee products :
2.3.Chemical properties and ATR-FTIR :

III- MATERIALS AND METHODS
3.1. Measurements of bee activity:
3.2. Biological assays
3.3 Chemical assays and ATR-FTIR

IV- RESULTS AND DISCUSSION
4.1. Honeybee activity in collecting and producing some bee products:
4.3. Chemical properties and ATR –FTIR of some bee products

VI- REFERENCES

I. SUMMARY

The present investigation was carried out at the department of bee research, plant protection institute, Sakha–Kafer EL-Sheikh province and department of pharmacognosy, faculty of pharmacy, Mansoura University during the period from the beginning of January 2014 to the end of December 2014.

The study includes measurement of bee activity in collecting and producing honey, propolis and bee venom according to bee race, in a comparative study between Carniolan hybrid and Italian hybrid. For one year and study of some biological and chemical properties of some bee products (Honey, propolis and bee venom) and their effects on three types of cancer cells (Hep-G2, Caco-2 and MCF-7) in a comparative study with the effort of two drugs (5Fu and Cis-pt) in different concentrations. The study dealt with the following points.

1. Activity honeybee in collecting and producing honey, propolis and bee venom according to bee race.
2. Biological properties of some bee products and their effects on some cancer cells.
3. Chemical properties of some bee products.

The obtained results can be summarized as follow:

I. Activity of honeybee in collecting and producing bee products according to bee race.

1.1 Honey:

1.1.1 Honey production:

- The average amount of clover honey yield/colony (9.57kg/colony) produced by Carniolan hybrid colonies was higher than that (8.30kg/colony) produced by Italian hybrid colonies.

- The amounts of cotton honey produced bya hybrid of Carniolan and Italian colonies were equal nearly and reached to 5.30 and 5.14 kg/colony, respectively.

- The average amount of clover honey per colony in a hybrid of Carniolan colonies ranged between 8.60 and 10.51 kg/colony, while in Italian hybrid colonies ranged between 7.28 and 8.99 kg/colony.

- In relation to cotton honey, the average of the honey amount per colony in Carniolan hybrid colonies ranged between 4.88 and 5.85 kg/colony, while in Italian hybrid ranged between 4.82 and 5.82 kg/colony.

1.1.2 Moisture and Total soluble solids:

- The highest moisture percentage was recorded from unripe clover honey. (Kafer EL Sheikh 2015) and ripe Brazilian papper honey; reached to 22.1% with total soluble solids 77.9%.

- The next highest moisture percentage recorded from ripe Banana honey, reached to 21.7% with total soluble solids 78.3%.

- The lowest highest moisture was recorded from rip Acasia honey, reached to 16.1% with total soluble solids 83.9%.

- The next lowest moisture was recorded from ripe clover honey (Asyut) and ripe Fennel honey reached to 17.1 and 17%, respectively.

- The highest total soluble solids were recorded from ripe Acasia honey, reached to 83.9% and from ripe clover honey (August), reached to 83%.

- The lowest total soluble solid was recorded from unripe clover honey (Kafr El-Sheikh) and rip Brazillian pepper honey, reached to 77.9% with moisture 22.1%.

1.1.3 Qualitative Microscopical analysis of Pollen in honey samples:

- Ripe clover honey Kafer Elsheik 2014 showed five plant spices of pollen grains were spread as followed, 45% Trifolium alexandrinum, 21% Medicago sativa, 28% Eucalyptus spp, 5% Phoenix dactylifera, 1% Fam.Mackinlayaceae.

- The Unripe clover honey Kafer Elsheik 2014(Trifolium alexandrinum) showed six plant spices of pollen grains were Spread as followed, 38 % Trifolium alexandrinum, 37% Medicago sativa, 3% Eucalyptus spp, 5% Phoenix dactylifera, 2% Casuarina spp, 15% Brassica rapa.

- While the ripe clover honey Kafer Elsheik 2015(Trifolium alexandrinum) samples showed six plant spices of pollen grains were Spread as followed, 65% Trifolium alexandrinum, 5% Medicago sativa, 15% Eucalyptus spp, 4% Phoenix dactylifera, 7% Casuarina spp, 4%Fam.Asteraceae.

- Unripe clover honey Kafer Elsheik 2015(Trifolium alexandrinum) showed seven plant spices of pollen grains were Spread as followed, 61% Trifolium alexandrinum, 13% Medicago sativa, 3% Eucalyptus spp, 7% Phoenix dactylifera, 5% Casuarina spp, 3% Cuminumcyminum, 8% Fam. Parsley.

- In Clover – Asyut honey (Trifolium alexandrinum) seven plant spices of pollen grains were Spread as followed, 77% Trifolium alexandrinum, 9% Medicago sativa, 1% Eucalyptus spp, 5% Phoenix dactylifera, 5% Fam.Mackinlayaceae, 2% Casuarina spp, 1% Medical plant.

- Clover (market honey) (Trifolium alexandrinum) showed seven pollen grains plant spices were Spread as followed, 45% Trifolium alexandrinum, 1% Medicago sativa, 19% Eucalyptus spp, 13% Phoenix dactylifera, 17%Fam.Mackinlayaceae, 4%Casuarina spp, 1%Fam.Asteraceae.

- The Banana honey showed ten pollen grains plant spices, were Spread as followed, 8%Trifolium alexandrinum, 1% Medicago sativa, 5%Eucalyptus spp, 42%Fam.Mackinlayaceae, 17%Medical plant, 7%Helianthus annuus, 3%Zea mays, 3%Fam.Cucurbitaceae, 7%Cyperus spp,7%Citrus spp.

- As in Acasia honey four pollen grains plant spices were found were Spread as followed, 93% Acacia spp, 5% Eucalyptus spp, 1% Phoenix dactylifera, 1%Ocimum basilicum,

- While the Fennel honey (Foeniculim vulgare) samples showed six plant spices of pollen grains were Spread as followed, 3% Trifolium alexandrinum, 5% Phoenix dactylifera, 60% Fam.Mackinlayaceae, 12% Casuarina spp, 3% Raphanus sativus,17% Pimpinella anisum.

- While the Sider honey (Ziziphus spp.) samples showed six different plant spices of pollen grains were Spread as followed, 1% Eucalyptus spp, 16% Phoenix dactylifera 75% Fam.Mackinlayaceae, 3% Casuarina pp, 2% Fam.Asteraceae,3% Nigella sativa.

- As the Brazillian papper honey (Schinus terebinthifolia) samples showed four different plant spices of pollen grains were Spread as followed, 2% Eucalyptus spp,5% Phoenix dactylifera, 3% Casuarina spp, 90% Schinus terebinthifolius.

- The Cotton honey (Gossypiumbarbadense) samples showed nine different plant spices of pollen grains were Spread as followed, 30% Trifolium alexandrinum, 3% Medicago sativa,11% Eucalyptus spp, 10% Phoenix dactylifera, 16% Casuarina spp,13%Fam.Asteraceae,1% Helianthus annuus,15% Zea mays, 1% Cyperus spp.

- The Medicinal plants honey, Coriander(Coriandrursativum)+Black cumin(Nigella sativa)+Anise (Pimpinellaanisun),(Gossypiumbarbadense) samples showed eight different plant spices of pollen grains were Spread as followed, 1% Eucalyptus spp,1% Phoenix dactylifera, 93% Fam.Mackinlayaceae, 1% Casuarina spp, 1%Fam.Asteraceae, 1% Fam.Cucurbitaceae, 1% Nigella sativa, 1% Brassica napus L.

- The Sunflower (Helianthus annuus)+Sesame(Sesamum indicum) samples showed six different plant spices of pollen grains were Spread as followed, 1% Eucalyptus spp, 9% Phoenix dactylifera, 2% Fam.Mackinlayaceae, 80% Helianthus annuus, 7% Zea mays, 1% Ocimumbasilicum.

- The Citrus honey samples (Citrus spp.) showed eleven different plant spices of pollen grains were Spread as followed, 1% Trifolium alexandrinum, 1% Medicago sativa,16% Eucalyptus spp, 8% Phoenix dactylifera, 6% Fam.Mackinlayaceae, 25% Casuarina spp,1% Medical planet,1% Vicia faba, 6% Fam.Asteraceae, 35% Citrus spp, 1% Nigella sativa.

1.2 Bee venom:

- The total amount of bee venom produced by Italian hybrid colony was 1.2719 gm/colony/year, while the total amounts of bee venom produced by Carniolan hybrid were 1.2509 gm/colony/year.

- The highest amounts of bee venom were produced in autumn season 0.3915 and 0.3728 gm/colony in hybrid of Carniolan and Italian race, respectively. Under 25C0 and 60% RH.

- The second highest season for produce bee venom was in winter 0.3377 and 0.3279 gm/colony in a hybrid of Italian and Carniolan race, respectively under 21 C0 and 65 % RH.

- The lowest amounts of bee venom were produced in spring season 0.2584 and 0.2139 gm/colony in a hybrid of Italian and Carniolan race, respectively reached to lowest percentage 20.32 and 17.10%, respectively and 30 C0 and 62% RH.

1.3 Propolis:

- The total amount of propolis gathered by Carniolan hybrid colony was 46.43 g/colony/year, while the total amount of propolis gathered by Italian hybrid was 20.76 g/colony/year.

- The highest amount of propolis were collected in summer and spring seasons. In summer were 20.12 and 7.23 g/colony in Carniolan and Italian hybrid; respectively while in spring were 13.69 and 5.68 g/colony, respectively.

- The lowest amount of propolis was collected during Winter and Autumn, in winter in Carniolan and Italian hybrid were 8.77 and 3.67 g/colony, respectively, while in Autumn were 4.35 and 4.18 g/colony, respectively.

- Carniolan hybrid colonies collected amounts of propolis more than Italian hybrid colonies.

- Carniolan hybrid colonies gathered 69.33% from the total amounts of propolis of two races, while Italian races colonies gathered 30.67% from the total amount of propolis.

1.4 Correlation coefficient between quantities of propolis and bee venom and both temperature and relative humidity:

1.4.1 In craniolan hybrid colonies:

- It was found a strong positive signified correlation between propolis and temperature, while it was found insignificant correlation between temperature and both relative humidity and bee venom and between relative humidity and both propolis and bee venom. Also between propolis and bee venom.

1.4.2 In Italian hybrid colonies:

- It was founduninsignificant correlation between temperature and both relative humidity, propolis and bee venom and between relative humidity and both propolis and bee venom. Also between bee venom and propolis.

2.Biological properties of some honey bee products (Honey - bee venom - Propolis):

2.1.Honey :

- The highest cytotoxic activity was 52.53 in breast cancer with citrus honey, while the lowest cytotoxic activity was 0.75 and 1.75 in liver cancer with acacia honey and clover (supermarket honey), respectively.

- The highest toxicity of liver cancer cells was 32.92 with sunflower honey and sesame, while the lowest cytotoxic activity was 0.75 and 1.75 with acacia honey and clover (supermarket honey), respectively.

- The highest toxicity of colorectal cancer cells was 49.84 with basil honey (Assiut), while the lowest cytotoxic activity was 25.53 with fennel honey.

- It was also observed that the highest activity of breast cancer cells was 52.53 with honey citrus, while the lowest activity of cytotoxic was 2.53 with acacia honey.

2. 2. Bee venom:

- The highest cytotoxic activity was 93.92 in liver cancer with both Italian and Carniolan venom (anatomy), while the lowest cytotoxic activity was 86.28 in breast cancer with Carniolan bee venom (in electric shock).

- It was found that the highest activity of liver cancer cells was 93.92 with the venom of Carniolan and Italian (anatomy), while the least activity of cytotoxic was 91.85 with the venom of Carniolan (in electric shock).

- The highest toxicity of colorectal cancer cells was 93.23 with Italian bee venom (anatomy), while the lowest cytotoxic activity was 91.76 with Carniolan toxin (in electric shock).

- It was also observed that the highest activity of breast cancer cells was 90.21 with Italian bee venom (in electric shock)., While the lowest cytotoxic activity was 86.28 with the venom of Carniolan (in electric shock).

- The study showed that the drugs used in the treatment of cancer 5FU and Cis-pt gave the lowest results in different concentrations (6.25, 12.5, 25, 100.50) compared to bee venom, where the highest toxic activity of cancer cells was 90.24 in colorectal cancer with Cis-pt100, While the lowest cytotoxic activity was 6.95 in liver cancer with 5FU-6.25.

2. 3. Propolis:

- The highest cytotoxic activity was 84.99 in liver cancer with Libyan propolis, while the lowest cytotoxic activity was 7.87 in liver cancer with Egyptian propolis.

- The highest cytotoxic activity of hepatocellular carcinoma was 84.99 with Libyan propolis, while the lowest cytotoxic activity was 7.87 with Egyptian propolis.

- The highest cytotoxic activity of colorectal cancer cells was 79.63, while the lowest cytotoxic activity was 27.73 with Egyptian propolis.

- It was also observed that the highest cytotoxic activity of breast cancer cells was 83.57 with Libyan propolis, while the lowest cytotoxic activity was 73.92 with Egyptian propolis.

- The study showed that the cytotoxic activity of the cancer cells of some of the main propolis components, Cinnamic acid, Ferulic Acid and Caffeic acid had the lowest results compared with propolis. The highest cytotoxic activity of the cancer cells was 30.32 in colorectal cancer with Ferulic Acid, in liver cancer with Caffeic acid and Cinnamic acidrespectively.

3. Chemical and ATR-FTIR spectral analysis (infrared) for some honeybee products:

3. 1. Chemical analysis:

3. 1. 1. Honey:

3. 1. 1. 1. total phenolic:

- The highest total phenolic was 121.628 mg / 100 mg in cotton honey, while the lowest value of total phenolic was 31.666 mg / 100 mg in citrus honey.

3. 1. 1. 2. Invertase enzyme :

- The highest value of invertase was 8.8 in the honey of medicinal plants, while the lowest value of the enzyme was 3.6 in the honey of clover (unripening 2014).

3. 1. 1. 3. Diastase :

- It was found that the highest value of diastase was 19.7 in the honey of medicinal plants, while the lowest value of the enzyme diastase was 8.3 in honey clover (unripening 2014).

Invertase(I)

The percentage between the enzyme Diastase and the Invertase should be greater than 0.5 in the fresh honey.Diastase(D)

- The highest observed percentage of I/D was 60, 59, 0.52, 0.52 in clover (Assiut), Sidr, Acasia, Citrus and Sunflower + Sesame respectively found it the most freshness.

3. 1. 1. 4. Catalase :

- The highest value of the catalase enzyme was 24.8 in the honey of medicinal plants, while the lowest value of the catalase enzyme was 14.5 in market honey.

3. 1. 1.5. Hydroxymethylfurfural :

- The highest value of hydroxymethylfurval was 6.8 mg / kg in the market honey, while the lowest value of hydroxymethylfurval was 1.4 mg / kg in both acacia and citrus honey.

3. 1. 2. Bee venom:

3. 1. 2. 1. Melittin:

- The highest value of Melittin was 68.12% in Carniolan venom (dissected method), while the lowest value for Melittin was 29.82% in Italian bee venom (an electric shock)

3. 1. 2. 2. Phospholipase :

- It was noticed that the highest value of Phospholipase was 15.37% in theCarniolan venom (dissected method), while the lowest value of Phospholipase was 9.65% in the venom of Italian bees (electric shock).

3. 1. 2. 3. Hyaluronidase:

- It was also noted that the highest value of hyaluronidase was 2.94% in the venom of carnivores (electric shock), while the lowest value of hyalurindrias was 0.91% in the venom of Italian bees (dissected method).

3. 1. 3. Propolis:

3.3.2.3.1.Total phenolic compounds:

- The highest value of total phenolic compounds was 121.958 mg / 100g in Bulgarian propolis, while the lowest value was 58.508 mg / 100g in Egyptian propolis.

3. 1. 3. 2. Caffeic acid:

- The highest value of Caffeic acid was 2.56 mg / ml in Bulgarian propolis, while the lowest value was 0.75 mg / ml in Egyptian propolis.

3. 1. 3. 3. Cinnamic acid:

- It was also noted that the highest values ​​of cinnamic acid was 30.8 mg / ml in Bulgarian propolis, while the lowest value was 14.5 mg / ml in Egyptian propolis.

3. 1. 3. 4. Ferulic Acid :

- It was noted that the highest value of ferric acid was 2.65 mg / g in Bulgarian propolis, while the lowest value was 1.25 mg / g in the Egyptian propolis.

3. 1. 3. 5. Antioxidant activity:

- The highest antioxidant activity was 38.7 mg / ml in Bulgarian propolis, while the lowest value was 11.23 mg / mL in Egyptian propolis.

3. 2. ATR-FTIRSpectrum analysis (infrared):

3. 2. 1. Honey:

- The study showed that there are groups of hydroxyl, sugar, carbonyl and groups of ethraolifes in ripening and unripeningclover honey 2015, clover (Assiut), citrus and market honey.

3. 2. 2. Bee venom:

- The study showed that, there are groups of carboxylic, amide group, carbonyl, an indicator of the presence of betide, etherolivatic groups and aromatic compounds (in the range 668 and 679 cm -1) in the Carniolan venom (dissected method), craniolan (electric shock), Italian bee venom (dissected method)and Italian bees venom (electric shock).

3. 2. 3. Propolis:

- The study showed that there are groups of Carboxyls, proteins, betides, carbonyl group, aromatic components and flavonoids in the range of 668 and 650 cm -1 in the Egyptian Propolis.

I. Introduction

Apitherapy is the art and science of making therapeutic use of products of honey bee, including honey, bee pollen, propolis, royal jelly, bees wax and bee venom. apitherapy is defined as api, meaning bee and therapy from the greek word therapia, which literally means “curing” or “healing.” apitherapy is healing by using bee products.hippocrates, the greek physician known as the "father of medicine", recognized the hialing virtues of bee venom for treating arthritis and other joint problems. Since the ancient egyptians, cultures across the ages have revered the honey bee for its healing powers.Integrative medical apitherapy is the application of bee products like honey, pollen, royal jelly and propolis in healing therapies.apitherapy is an ancient alternative healing modality dating back several thousand years and used by hippocrates, mentioned by plato, and others throughout our history.

Honey is a natural product containing a complex mixture of sugars,minerals, proteins, vitamins, organic acids, flavonoids, phenolic acids, enzymes and other phytochemicals. It contains antioxidant molecules such as flavonoids, phenolic acids, catalase, carotenoids, peroxidase and catalase (Gheldof and Engeseth, 2002). Documented biological activities of honey includes antioxidant, immuno-modulatory, cancer prophylactic and curative properties (Al-yahya et al., 2013). In addition, experimental evidence indicates that honey from variety of floral and geographical sources could exert several health-beneficial effects, whichincludes gastroprotective, hepatoprotective, reproductive, hypoglycemic, antioxidant, antihypertensive, antibacterial, anti-fungal and anti- inflammatory effects (Erejuwa et al., 2012). Honey contains about 200 substances including sugars, phenolic acids, flavonoids, amino acids, proteins, vitamins and enzymes (Wang and Li, 2011). Phenolic compounds are considered among the main constituents contributing to the antioxidant and other beneficial properties of honey (Stephens et al., 2010; Aljadi and Kamaruddin, 2004). Studies show that the great majority of the bioactive compounds in honey consist of molecules with phenolic structures, such as phenolic acids, flavonoids, procyanidins and anthocyanins (Küçük et al., 2007; Sahin et al., 2011; Tezcan et al., 2011).

Bee venom (BV) therapy has been used to treat different ailments. The main component and the principal toxin of BV, which constitutes approximately 50% of its dry matter, is melittin (MEL) (Gajski and Garaj-Vrhovac, 2013; Garaj-Vrhovac and Gajski, 2009; Orsolic, 2012; Son et al., 2007). Several mechanisms of MEL toxicity have recently been reported on a variety of cancer cells. MEL exerts toxic effects by damagingcell membranes and DNA, eventually causing apoptotic or necrotic cell death (Gajski and Garaj-Vrhovac, 2013; Orsolic, 2012; Son et al., 2007). Bee venom is a natural toxin produced by the honey bee (Apis mellifera), and has been used as a traditional medicine for treating various diseases, such as, arthritis, rheumatism, cancerous tumors, and various skin diseases (Billingham et al., 1973; Son et al., 2007). Bee venom contains a large number of biologically active peptides, including melittin, apamin, adolapin, and mast cell-degranulating peptide (MCCP) (Kwon et al., 2002). Many previous studies have examined the biological and pharmacological activities of melittin such as the anti-bacterial, anti-viral, anti-inflammatory, and anti-cancer effects (Boman et al., 1989; Jo et al., 2012; Park et al., 2008). Honeybee venom possesses diverse biological and pharmacological activities (Son et al., 2007). Its effectiveness has been demonstrated in treating pathological conditions such as cancerous tumors (Russell et al., 2004; Putz et al., 2006). BV has anti-cancer activity (Liu et al., 2002). Melittin, a major polypeptide of BV, is thought to function as a lytic agent that has been used traditionally against chronic inflammation and cancer (Huh et al., 2012) and also used for the therapeutic agent of arthritis, rheumatism, atherosclerosis, and cancer in traditional medicine (Terwilliger and Eisenberg, 1982; Son et al., 2007).

Propolis, a natural honeybee product, has been extensively used as a folk medicine (Castaldo and Capasso, 2002) and in food and beverages to improve health and prevent diseases (Banskota et al., 2001). Recently, propolis has received the special interest in the areas of oncology research as a source for prevention and treatment of cancer. Accordingly, a large number of compounds possessing the anticancer activity such as phenethyl caffeate (CAPE) (Lee et al. 2005; Xiang et al. 2006) been reported from propolis. Its chemical composition is very complex, over 300 constituents have been found in it, such as polyphenols (flavonoids, phenolic acids and their esters), terpenoids, steroids and amino acids, among which polyphenols have been considered as the primary biologically active compounds (Bankova, 2005). Phenolic compounds such as flavonoids, aromatic acids and diterpenes are the main components that account for the numerous biological activities of propolis (Galal et al. 2008), which include antioxidant, antifungal, antiviral, antimutagenic and immunomodulatory activities (Ansorge et al., 2003; Jasprica et al., 2007;Paulino et al.,2008). There are a number of review articles on the anti-cancer and anti-inflammatory activity of propolis, which also include a discussion on individual components in Poplar propolis, particularly Chrysin and Caffeic acid phenethyl ester (CAPE) and also polyphenols found in honey and propolis, and polyphenols in general (Araújo et al., 2011; Watanabe et al., 2011). CAPE has been extensively investigated for its anti-cancer activity, particularly colon cancers (Borrelli et al., 2002; Rao et al., 1993; Liao et al., 2003; Xiang et al., 2006).

The aim of this study is to integrate the expertise of both bee research - plant protection research institute branch of Sakha - Kafr El-sheikh governorate and Department of Pharmacognosy, Faculty of Pharmacy, Mansoura University to investigate the following:

1- Honeybee activity in collecting and producing (honey, venom and propolis)according to bee races.
2- Biological properties of honeybeeproducts (honey, venom and propolis) and their effects on somedifferent cancercalls.
3- Chemical properties and ATR-FTIR of somehoneybee products (honey, venom and propolis).

II – REVIEW OF LITERATURE

2.1. Honeybee activity in collecting and producing some bee products:

2.1.1. Honey

2.1.1.1. Honey production :

Shorert and Hussein (1993) found that coloniesfed with sugar syrup mixed with a protein supplement were given 100 g supplement produced significantly more honey than control colonies (fed only on sugar syrup). They noticed that soybean flour had the greatest effect on honey yield (14.3 kg/colony), followed by agwa date (13.0 kg/colony), pollen (12.0 kg/ colony) and chick pea (11.8 kg/colony).

Abbas et al. (1995) concluded that first group contained an average of 7.12 kg honey/box, where as colonies fed on the black gram diet had 8.62 kg/box. Colonies received no pollen substitute had only 1.87 kg honey/box. They used black gram (phaseolus mungo) 550 g/ kg diet instead of soybean meal as a substitute for pollen in the diet of honeybees (Apis mellifera L.) for three months in the summer rainy season.

Szymas and Przybyl (1995) indicated that colonies stored higher honey with feeding on pollen substitutes made from potato protein (32 %) ground soybeans, yeast, powdered skimmed milk, powdered hen's eggs, extruded maize and vitamins.

Kumova (1999) demonstratid that colonies fed on 1:1 sucrose-water + vitamins + minerals + antibiotics was the highest yield of honey 43.80 kg/colony, followed by 36.30 kg/colony from colonies fed on syrup (1:1 sucrose : water) and 19.20 kg/colony from colonies without feeding.

Mladenovic et al. (1999) showed that greatest honey area was recorded in the group additionally fed on syrup with yeast (37.8 % higher than the control) in the spring.

Taha (2007) stated that produced honey exceeded those in the apiary of the Faculty of Agriculture, Kafr El-Sheikh University by 100 (4.56kg/colony) with colonies located in banana farm during August and September (the peak of blooming period of banana Musa sp.)

Sande et al. (2009) found that the honey yield doubled in hives located less than one kilometer from the forost, but decresed at 3 Km proximity. honey yield per harvest (kg) and obtained samples from hives placed at varying distances from Arabuko Sokoke Forest (ASF). All the honey samples met internationally required quality standards, although sugar levels were at the lower limit.

Taha et al. (2009) found that, inKafr El-Sheikh region the highest mean honey yield (3.10 & 3.09 kg/colony) was recorded during clover blooming period in the two years respectively. They noticed that the highest mean honey yield(kg.)/colony (4.45 & 4.54) was recorded during citrus blooming period in Rasheed region, followed by clover blooming period (3.68 & 3.71 kg/colony) in Ammya region in the first and second years respectively.

Al-Ghamdi et al. (2016) assessed the appropriatecolonies number of honeybee in areas occupied by ziziphus spina-christi treas and Acacia tortilis shrubs at Al-Baha region,Kingdom of Saudia Arabia. Average numbers of bee colonies were 530 and 307 per square Kilometer. Numbers of foragers were 0.55 and 11.12 to ziziphus trees and Acacia shrubs respectively. The produced honey was 5.21 and 0.34 Kg per ziziphus and Acacia plants respectively. The study revealad that the colonies were overcrowded in relation to the sources of nectar.

2.1.1.2. Moisture and Total soluble solids (TSS):

De Bruijn and Sommeijer (1997) presented their first results on the composition, properties and antibiotic activity of honeys of different species of Melipona and compared them to those of Apis. They found that honey of M. favosa and M. trinitatis contained on average 23.5% moisture (N = 28).

Bijlsma et al. (2006) found that the moisture content (MC) value in honey from Plebeia tobagoensis (42.0%). The other MC values were 36.2% for Trigona nigra, 31.2% for Melipona favosa and 32.2% for Melipona trinitatis. The lowest MC was found in honey of Apis mellifera (20.2%). There was little variation between colonies of the same species at the same site, but honey of M. favosa from Trinidad had a higher MC (35.1%) than that from Tobago (30.2%). The finding that the MC of honeys of stingless bee species varies according to the species and to the area where it is produced.

Chirife et al. (2006) stated that correlation between water activity (major factor in preventing or limiting microbial growth and determining the type of microorganisms encountered in food) and % moisture in Argentine honeys was then experimentally determined and explained on the basis of the above analysis. They pointed that a very good straight line relationship (correlation coefficient 0.971) was found between both parameters in the range examined (15– 21% moisture), and also the goodness of fit of the regression equation was found to be quite satisfactory.

Yanniotis et al. (2006) measured viscosity of honey in two honeydew honeys (pine and fir) and four unifloral nectar honeys (thymus, orange, helianthus and cotton) at their initial moisture content as well as at 17%, 19% and 21% water content at 25, 30, 35, 40 and 45 C.They suggested that The initial moisture content of the samples varied from 15% to 17.1%. Moisture content of honey usually varies from 14% to 18%, but it must not exceed 20% according to the Greek law. They found that effect of moisture is more pronounced for moisture content up to about 19%. They showed above this level the effect is weak. They noticed viscosity of pine and fir honey was found to be substantially higher than the viscosity of thymus, cotton, helianthus and orange honeys at the same temperature and moisture content especially at the lower end of the moisture and temperature range tested (15–21% moisture and 25–45 C).

Farag, (2007) in Egypt, showed that water content the honey sample collected lower Egypt ranged between (19.0) and (24.5%) with the mean of (21.77%) while in samples collected from middle Egypt ranged between 19.5 and 21.5 % with the mean 20.33%, the moisture of clover honey collected from Lower Egypt ranged between 18.0 and (19.5%), with the mean of (18.63), while the moisture in clover honey collected from middle Egypt ranged between (18.0) and(20.5%)with the mean of (19.25%).

Abramovic et al. (2008) found that water content in honeydew honeys ranged from 13.4% to 18.0% and in flower honeys, the water content was between 14.0% and 18.6%. They noticed that statistically significant linear correlation between water activity (aw) and the water content of honeys was found. In honeydew honeys, the water activity at the same water content was higher than in flower honeys.

Eissa et al., (2010) in Egypt, showed that the total soluble (TSS) from (80.38 – 82.75) with mean (81.00%) of clover honeys.

Metwaly, (2011) in Egypt, stated that the total soluble solids (T.S.S) value of honey samples of the three Egyptian honeys (citrus and clover) was from (80.00 – 82.75%), the minimum value of (T.S.S) was detected in citrus honeys, while the maximum was detected in the clover honeys. The (T.S.S) of citrus honeys samples collected from Egyptian regions was ranged from (80.00-82.00%) with the mean of 80.95% considering the clover honey, the (T.S.S) range was from (80.75 – 82.75 %) for samples collected from 11 Egyptian regions with a mean of (81.06) %.

Escuredo et al. (2013) indicated that moisture content is one of the most important characteristics, influencing physical properties of honey such as viscosity and crystallization, as well as other parameters: color, flavor, taste, specific gravity, solubility, and conservation. They evaluated the water content of 187 honeys harvested in Northwest Spain and the sample's contents ranged from 16.9 % to 18.0 %, averaging 17.6 % .

Yücel and Sultanoğlu (2013) showed that water is the second largest constituent of honey. Its content may vary from 15 to 21 g 100 g-1 depending on the botanical origin of the honey, the level of maturity achieved in the hive, processing techniques and storage conditions.

Karabagias et al. (2014) found that water content of 39 pine honey samples in Greece, the samples showed values between 10.50% and 20.50%. The percentage of moisture in honey can also vary in regions with high relative humidity, or depending on the season, as honey is more likely to suffer a fermentation process in the rainy season rather than the dry season. They noticed moisture in honey can also increase during the processing operations of the product, as well as the inadequate storage conditions, because honey is hygroscopic and absorbs moisture from the atmosphere.

Nafea et al., (2014) in Egypt, results of physical analysis showed that the viscosity and total soluble solids (TSS) in all honey types were ranged between from (13.6 – 87.8), (77.0 – 83.2 %), respectively .

2.1.1.3. Quantitative microscopical analysis of pollen in honey:

Tsigouri et al. (2004) In Greecefound that 208 samples were unifloral with 178 of them representing the main types of unifloral honey produced in Greece; that is fir, pine, chestnut, cotton, orange and thyme honey. The pollen types identified in these honeys ranged from 11 to 45%. Chestnut nectar honey contained w90% chestnut pollen, had a total number of plant elements of w245,000/10 g, and low pollen diversity. Cotton honey contained 1.2 to 16.5% cotton pollen, belonged to Maurizio’s Class II, and had 22 pollen types, with Castanea sativa L. present in all samples. Orange honey contained 2.9 to 26.5% Citrus spp. pollen, belonged to Maurizio’s Class II, and was characterized by the presence of Brassicaceae, Fabaceae, Olea europea L., Quercus coccifera L. and Rosaceae. In thyme honeys Thymus capitatus Hoffm. & Link. pollen was secondary or predominant ranging from 18.3 to 69.3%. These honeys belonged to Maurizio’s Classes I or II and contained greater than 30 pollen types. Other Lamiaceae, Hypericum spp., Brassicaceae, Fabaceae, Rosaceae, and Cistus spp. pollen types appeared in the greatest number of thyme samples.

Sodre et al. (2007) In Brazilnoticed that identify the pollen types occurring in 58 samples of honey produced in two states of the northeastern region of Brazil, Piauí (38 samples) and Ceará (20 samples), and to verify the potential of the honey plants during the months of February to August. They showed samples submitted to both a qualitative and a quantitative analysis. The dominant pollen in the State of Ceará is from Mimosa caesalpiniaefolia, M. verrucosa, Borreria verticillata, Serjania sp., and a Fabaceae pollen type, while in the State of Piauí it is from Piptadenia sp., M. caesalpiniaefolia, M. verrucosa, Croton urucurana and Tibouchina sp. They reported that quantitative pollen analysis detected five different pollen types occurring as dominant pollen in the 20 samples of the State of Ceará that were analyzed: Mimosa caesalpiniaefolia (Mimosaceae) (50.0%), M. verrucosa (Mimosaceae) (5.0%), Borreriaverticillata (Rubiaceae) (10.0%), Serjania sp. (Sapindaceae) (5.0%), and Fabaceae type (Fabaceae) (5.0%). different pollen types were also identified as dominant pollen in the 38 samples of the State of Piauí: Piptadenia sp. (Mimosaceae) (68.4%), M. caesalpiniaefolia (Mimosaceae) (5.3%), M. verrucosa (Mimosaceae) (7.9%), Croton urucurana (Euphorbiaceae) (2.6%), and Tibouchina sp. (Melastomataceae) (2.6%).

Feás et al. (2010) investigated that some properties of artisanal honey samples (n = 45) collected from the Northwest of Portugal by using different honey analysis tests. 77.8% of the total exceeded the quality parameters and should be labeled. All of the samples showed an Erica sp. pollen percentage P15%, and 42% of the total were monofloral Erica sp. They showed that occurrence frequency of the 21 pollen types identified from the 45 studied samples. The Fabaceae and Rosaceae families provided the greatest number of pollen types with 8 (Acacia, Cytisus, Chamaespartium, Genista, Lotus, Medicago, Trifolium and Vicia) and 3 (Prunus, Pyrus and Rubus) pollen types each, respectively. Rubus and Trifolium are present as IP (minor pollen (1–3%)) in 33 and 23 samples, respectively, corresponding to 73% and 51% ot the total analysed samples in percentages, and as SP (secondary pollen (16–45%)) Rubus and Trifolium are present in four samples, respectively. Erica sp. pollen is present in all honey samples, as PP (predominant pollen (>45%)) (in 20 samples) and as SP (in 25 samples, corresponding to a 55% of the total honeys). Next, the Eucalypthus pollen type is present as PP in one sample and as SP and IMP (important minor pollen (3–15%)) in 26 and 9, honeys respectively. Monofloral honeys are made up of nectar belonging to a single plant in an extent of at least 45%. They indicated that 42% of all the samples were monofloral Erica sp. honey samples with an Erica sp. pollen percentage higher than 35% could be labeled with a specific ‘‘Mel de Urze” or ‘‘Mel de Queirós” denomination. All of the samples analyzed in the present study, have an Erica sp. pollen percentage higher than 15% and only eight samples fall below 35%.

Ramirez-Arriaga et al. (2011) mintined that melissopalynological analysis of 39 honey samples from Oaxaca, Mexico, a total of 64 taxa belonging to 29 families were recorded. They characterised subtropical honeys by their botanical origin as follows: (a) monofloral honeys of Bursera simaruba, Clethra mexicana, Cordia alliodora, Lonchocarpus sp., Mangifera indica, Miconia argentea, Orbignya cohune and Quercus sp.; (b) bifloral honeys with an association of Heliocarpus donnell-smithii and Ceiba sp., Lonchocarpus sp. and Mimosa pudica, H. donnell-smithii and Mangifera indica, Miconia argentea and Miconia tenuiflora; (c) oligofloral honeys of Asteraceae; and (d) multifloral honeys with three or four species ≥10%. Monofloral honeys were placed in classes I, II, III, IV and Vaccording to absolute quantity of pollen grains, 21%(6757–17 406),46% (22 497–98 076), 28% (108 757–443 987), 3% (795 163) and 3% (1 049 609)respectively. Oligofloral (when two or more secondary taxabelonged to one botanical family) were class II, bifloral (when two pollen types had secondary percentages) were classes I and II, and polyfloral (when three or more pollen types were registered withsecondary percentages) honeys were assigned to classes I, II and III. Honey samples of Apis mellifera had a diversity index range of 0.3 to 2.7.

Song etal. (2012) demonstratid that qualitative and quantitative melissopalynological analyses for 19 Chinese honeys were classified by botanical origin to determine their floral sources. A diverse spectrum of 61 pollen types from 37 families was identified. Fourteen samples were classified as unifloral, whereas the remaining samples were multifloral. Bee-favoured families (occurring in more than 50% of the samples) included Caprifoliaceae (found in 10 samples), Laminaceae (10), Brassicaceae (12), Rosaceae (12), Moraceae (13), Rhamnaceae (15), Asteraceae (17), and Fabaceae (19).They found in the unifloral honeys, the predominant pollen types were Ziziphus jujuba (in 5 samples), Robinia pseudoacacia (3), Vitex negundo var. heterophylla (2), Sophora japonica (1), Ailanthus altissima (1), Asteraceae type (1), and Fabaceae type (1). The absolute pollen count (i.e., the number of pollen grains per 10 g honey sample) suggested that 13 samples belonged to Group I (,20,000 pollen grains), 4 to Group II (20,000–100,000), and 2 to Group III (100,000–500,000).

Costa et al. (2013) suggested that honey samples from apiaries located in the province of Catamarca Argentina,a total of 86 pollen types, belonging to 39 families, were identified from the honey samples and 68 (79%) were native flora. Ten types were classified as predominant pollen types and were present in the monofloral honeys (n = 18), Prosopis spp., Cercidium praecox, Larrea divaricata and Tournefortia lilloi are those with the highest frequency. Pollen from Fabaceae and Asteraceae has the greatest representation in honey, with 14 and 11 types, respectively. Most of the honey samples correspond to groups II (normal pollen grain content, 39%) and III (rich in pollen grain content, 30%). They pointed that honeys analysed had a typical pollen array of arid Chaco and southern Andean Yungas flora.

Puusepp and Koff (2014) concluded that the pollen content of 325 honey samples was analysed with an average of 400 pollen grains counted in a sample. The floristic spectrum of plants and on the identification of the most common and important plant sources for honey. They observed that more than 120 pollen types were identified in the examined honey samples. The pollen of Apiaceae, Fabaceae, Asteraceae, Poaceae, Fagopyrum esculentum, Frangula alnus and Calluna were present in more than 25% of samples. Typical Estonian honey is polyfloral, the average number of species is 13 taxa per sample. They found that the concentration of pollen grains per gram of honey varies from 100 to 700 000. The current information provides new insights into the pollen composition of Estonian honey and could be used to develop analytical standards for the pollen content of Estonian honey. They indicated that the proportion ofRosaceae pollen type and Calluna has decreased during the study years 2000–2010 (from 30% to 16%and 6% to 2%, respectively) and has been replacedby Brassicaceae and Salix pollen (from 19% to 40%and 6% to 20%, respectively).

2.1.2. Bee venom:

Morse and Benton, (1964a) stated that,bee venom collection from Africanized honeybees or the more defensive races in other parts of the world by standard electro-shock method cannot be recommended. Theyfound that mass reaction of Africanized honey bees may result in contamination of the collected venom. Colony arousal can become so overwhelming that bees start killing each other and alert other colonies or attack the beekeeper and bystanders. Theyshowed that venom is collected by this method in Brazil and Argentina, with only minor modifications. However, Morse and Benton (1964b) found no such evidence for reduced productivity.

Gunnison (1966) indicated that used a standard electro-shock collecting apparatus with the cooling system in order to preserve more of the volatile compounds.

Mitev (1971) suggested that bee venom which collected every three days produce 14% less honey. Even European colonies remain disturbed for up to a week.

Suzuki (1970) indicated that alkaline glands is an opaquely whitish, minutes organ by contacting directly with air.pointed that Alkaline gland produces acid phosphatase being highly active in acid media of pH 5.6 - 6.0 without secreting alkaline phosphatase. Claimed that organ becomes slightly sticky and easy to break, and its function is lubricant for moving lancet, a neutralizer of remains of acid secretions and an adhesive for sticking the eggs to the com chamber.

Gary, (1974) indicated that the venom apparatus of the honey bee, like that of many social insects, has a prime role of defense to the colony and stinging behavior is most commonly observed in the proximity of the hive or nest. Concluded that Pheromones secretion is considered to be one of the main stimuli for inducing an aggressive attitude amongst defending worker bees.

Hsiang and Elliott, (1975) suggested that, electroshock method andvenom collected from surgically removed venom sacs showed different protein contents from this collecting method.

Pence, (1981) stated that,different extraction or collection methods of bee venom.found result in different compositions of the final products. Whoever venom collected under water avoid evaporation of very volatile compounds seems to yield the most potent venom.

Croft, (1988) concluded that, the production of potent bee venom requires good nectar, honey and pollen sources. Consequently, bees have more potent venom during the summer.

Hider, (1988) found that, a higher Venom production during summer months in which there is a peak of activity in the colony and when the relatively young individuals are beginning their defense behavior.

Lensky, and Cassier, (1995) observed that, bee sting apparatus and its associated glands, including the Koschewnikow glands, sting sheaths and setaceous membrane. They showed that more than 40 compounds have been identified in extracts of the worker sting apparatus, and 6 major compounds are releasers of alarm behavior.

Kaviani et al., (1995) concluded that,designed an electrical shock device on a new method of milking Apis mellifera for venom, consisting of 2 min intervals between each period of shock. They found that total of 9203 mg of dried honeybee venom (HBV) was collected during spring and summer from 8 hives in Iran. They noticed that purity percentage of collected HBV was 2.8%.

Rybak et al., (1995) indicated that, techniques used for honeybee venom collection using several types of devices. They reported that apparatus developed consists of an electro stimulator (generator) which passes current through electrodes mounted every 5 mm in venom collecting frames fitted in one of the hive bodies. They found frames include a glass screen on which the venom is deposited. They pointed to venom collection was carried out every 14 days, for lh (early morning, before bee flight) or 2h(when foraging was occurring), with the collection frames in the upper body. They found Mid-July was the best period for venom collection.

Skubida et al., (1995) suggested that,compared the amount of venom collected when venom collecting frames were inserted (1) in the lower hive body, or (2) in the upper hive body, or (3) in an empty body placed between the upper and lower bodies. They Opined that fourth technique involved a super with a fixed set of 6 venom collecting frames (incorporating removable glass plates for scraping off the venom). They reported that,venom collection had no adverse effects on colony strength, brood rearing and productivity of honey (pollen and bees wax). They showed affected wintering performance, with colonies in group (3) most affected, and those in group (1) least affected. They demonstrated that colonies in group (2) gave the best results for total colony productivity.

Simics, (1995) suggested that a modem frame, which is placed on top of the frames in a hive when collectors have been put in each hive, 20 - 40 are connected together and electric impulses are passed through for 30 min. They found that venom, which is scraped from the device in dried form, is claimed to be uncontaminated. They rported that,Colonies are relatively unaffected by procedure, an observation during the collection period showed that, on average 68 bees died per colony.

Asmaa Anwar, (2000) reported that production of bee venom ranged between 33.5 - 37.0 mg during spring season and 30.0 - 35.0 mg during summer season. Found That Italian bees have the heavies while, the Craniolan have the lowest amount of bee venom production. showed mean weights were 37.0, 34.0 and 33.0 mg during spring for Italian, Midnite and Craniolan, respectively. These means were 35.0, 30.0 and 30.0 mg during summer.

Simics, (1995a) indicated that, the quality of collected bee venom (BV) is determined by several factors such as: honey flow, chronological âge of the bees in the hive, weather condition, venom collecting technology and the technical parameters of the collector device. The device should not kill more than 5-15 bees per hive during 30 min. of collecting time.

Nentchev, (2001) come to the conclusion that,Bee venom was obtained through electro stimulation twice a month using the schedule 30 min stimulation and 60 min pause. He noticed that, the annual yield of bee venom from a bee colony for 15 sessions at 14-day intervals between March and October was 3.804 g. showed most bee venom was obtained in June and July, and the least in March and October.

Mohanny, (2005) come to the conclusion that, the general yearly amount of bee venom collected by the two honeybee hybrids was 2.8237 g/colony. He found that According to different months of the year, the collected quantities fluctuated from 0.8387 g/colony in August (the greatest) to 0.0386 g/colony in December (the lowest). He reported that,According to the different seasons, the collected quantities could be arranged descending as follows; summer, spring, autumn and winter.

Al-Sharawy, (2008) stated that,collected honeybee venom all the four seasons of the year. They showd that, spring season recorded the highest in bee venom collection weights between the other seasons and the winter season was the lowest in bee venom quantity. They found that, the best seasons to collect the largest quantity of the bee venom are the spring followed with the summer season.

2.1.3. Propolis :

Starostensko (1968) indicated that, some races of honey bees collect propolis more active than others. They pointed that Grey mountain Caucasian bee collect rather more than dark forest bees, whereas Italian, Ukrainian and Far East bees collect very little. While Czechoslovakia, A. mellifera carnica bees are most widely spread,' it produces less propolis that native bee under the conditions in this country, A. mellifera caucasia produces more propolis.

Spangler and Taber (1970) reveald that, propolis is soft and sticky at warm temperatures and can be molded to fill holes and gaps or spread over surfaces. They noticed that at cool temperatures and as it ages, propolis becomes brittle and hard.

Krupicka (1972) suggested that, some races of honey bees actively collect propolis than others. They opined that Grey mountain Caucasian. A. mellifera Caucasia bee collect rather more than dark forest bees, whereas Italian Ukrainian and Far East bees collect very little.

Taber and Barker, (1974) showed that, Propolis is gathered by bees from trees and other vegetations, either from buds (such as poplar) or from bark (particularly conifers).they found that bees may collect resinous substitute materials such as chalking compounds.

El-Sarrag (1977) showed that, Sudanese bees and their hybrids collected more propolis during the summer (9.0-12.7g) than in the winter (2.5 - 9.0 g). Jachimowicz (1978) reveald that the bees collect propolis on warm days with temperature of more than 20°c and only from 10am to 3 pm especially in latesummer and in autumn.

Jachimowicz (1978) suggested that, it is possible dung harvesting that some mandibular sécrétions may be of the labial glands. Penetrate propolis as it is in the case of wax (in order to be better mixed). He indicated that, the amount of propolis collected by the bees depend on the characteristics of the place where bees are located on the climatic conditions.

Mizis (1978) reported that, production of propolis in this State was about 1.5, 2, 3, and 2.5 and from 3 to 3.5 tons at 71, 72, 73, 74 and 1978, respectively. They observed that, in one work-day a beekeeper can collect from 1 to 1.5 kg propolis. He pointed that each colony irrespective of race and method of collection can produce from 50 to 700 g propolis, and that with the Caucasian Mountain Grey bees, 2-3 times more propolis was obtained.

Caillas, (1978) demonstratid that, Propolis is a natural bee product and according to propolis has a two-form origin:

1. An internal origin Propolis might be a resin residue coming from the first phase of pollen digestion in a small organ placed between the sac and lower gut of bees. All comb cells, and especially the newly built one, are varnished with this internal propolis before the queen lays eggs them. He observed that,the greatest quantity of propolis produced by the bees seems to have this origin. It recognized under the microscope because of the pollen grains it contains.

2. An external origin it was believed that the forager bees harvested propolis as it is from trees buds, particularly from poplar and alder, but it has been 6 found that they harvest it also from other trees.

Ghisalberti, (1979) concluded that,honey and other hive products have been utilized in nutrition and remedy since ancient times and regarded as a symbol of power and health in folk remedy.Noticed that natural product derived from plant resins and collected by honeybees to seal the walls and entrance of the hive and contributes to protect the colony against different pathogens.mintined that In the north temperature zone, the propolis is collected from different plant sources, particularly species of poplar, Birch, Alder, Peach and Horse chestnut trees. He found that Western Australia, bees may collect some of its needs of propolis from grass trees, when the corbicula have filled, the bees finally deliver the propolis to the hive. The bees might have to wait on the wall of the hive-records ranged from one hour to two days before the propolis load is removed by other bees, which immediately utilized it.

Ayoub (1982) come to the conclusion that, the lowest amount of propolis was collected during winter season. They pointed total amount of propolis gathered in winter average 6.3 g per colony representing 15.2% of the total yields. They found that in spring the collected amounts of propolis was average 10.4 g/colony, representing 26.3% of the total yields. They proposed that total amount collected during summer was average 14 65 g/ colony, about 37.0 % of the total yield. Autumn season was 8.5 g/ colony, representing 21% of the propolis production/ year.

Malkov and Sadovnikov (1985) concluded that, to increase propolis collection, strong colonies are needed and an availability of suitable woodlands. They showed that best months are May, June and August, and fine cloth (jute in best) should be used to collect the propolis from the hive, whereas only a few hives are involved, special frames can be used. They opined that cloths should be turned through 90 once or twice a week to prevent blockage of the flight holes. Nectar gathering in central RSFSR is not affected by propolis collection. They reveald that, average amount of propolis varies from 20 to 40 g per cloth. Theyreported that, bees most active in propolis gathering are Caucasian. They showed that, Central Russian bees also actively gather propolis. They found some colonies that collected much propolis (200 - 300 g/season), whereas others collected propolis hardly at all.

Battagiini et al., (1987) demonstrated that, process of collection of propolis by bees takes a long time and it may be interrupted by visits to the hive for feeding. They observed that, amount of propolis collected depends on the colonies strength, the seasonal period and zone.

Ashour (1989) come to the conclusion that, the amounts of propolis collected to increase during the warm and hot seasons (1.4-3.9 g/hive/ month) could be obtained during (May and September) , while (0.185 - 0.582 g/ hive/ month) were obtained in cold weather (December/ Januaiy)

El-Shaarawy (1989) indicated the amount of propolis collected by two hybrids (FI Italian and FI Camiolan). They found that FI Italian bees gathered 72.29 g /colony /year, while FI Camiolan bees gathered only 46.60 g /colony /year. They noticed that highest amounts of propolis were gathered during July and August and lowest in December and January.

Rybak et al. (1992) mintined that, the seasons of collection (Spring, Summer, Autumn) and the strength of the colony had significant effects on the contents of both types of impurities. They noticed that tendency of a colony to founding propolis produced in summer and autumn by strong colonies that collected much propolis.

Donia (1994) showed that, the mean quantity of propolis during 1990, 1991 and 1992 in winter seasons were 5.63, 5.30 and 5.20 g/ colony, respectively with mean 5.37 g/colony. He noticed in spring seasons the mean amount of propolis was 10.6, 11.4 and 11.5 g/colony, respectively with mean 11.16 g/colony. He observed that, in summer the mean of propolis were 12.8, 16.8 and 18.3g respectively, with mean 15.6g/colony and in autumn the mean amount of propolis were 9.6, 10.9 and 10.7 g/colony respectively with mean 10.3 g/colony.

Ghazala (1998) demonstratid that, Carniolan hybrid colonies (FI) collected monthly amounts of propolis significantly more than each of Carniolan honey bee and Carniolan hybrid (F2) colonies all over the year months of (1993, 1994 and 1995). They found that mean amounts of propolis were 9.41, 6.25 and 5.36 g/colony for the three strains, respectively.

de Jager et al. (2001) stated that, the Propolis quality and production was significantly higher in hives with the traps producing 75.6 ± 1.5 % resin and 361.87 ± 0.18g propolis compared to 64 ± 1.25 % resin and 36.1 ± 0.18g propolis in the control group. They noticed that hives containing propolis traps were more profitable than the control group when honey and propolis income were pooled (385.09 ± 0.40 Rand vs 324.04 ± 0.42 Rand).Therefore the increased propolis production improved profitability of the hive without affecting overall hive productivity. They found that maximum propolis (13.4 g and 192.8 g) and honey production (14.03 g and 16.3 kg) was obtained at 270 days for both control and treatment groups, The amount of honey produced increased and it seems that propolis production has a positive effect on honey production. Propolis production did not adversely affect hive size over the period.

Bradbear (2003) noticed that, gums and resins that bees gather from plants for propolis are the very substances exuded by plants for their own protection and healing.

El-Morsy (2003) reported that, the amount of collected propolis was increased by increasing the temperature through spring and summer seasons. He showed that collected amount every month was nearly 1.2 - 3.4 g/hive through summer season, while in winter the obtained amount was about 0.186 - 0.656 g/hive.

Mohanny (2005) indicated that, factors affecting quantities of collected propolis. he found that general yearly mean amount of propolis collected by bees was 76.07 g/colony. According to the different seasons, the collect quantities could be arranged discerningly as follows: summer, spring, winter and autumn with quantities of 33.458, 30.494, 7.240 and 4.885 g/ colony, respectively.

Ghazala (2006) concluded that, the effect of different seasons during the year on the collected amounts of propolis. They pointed that General seasonally mean was 23.428 g / colony. The highest amounts of propolis were gathered during autumn which were 37.55 and 19.83 g/colony for Fj and F2, respectively. He showed that significant differences between seasons during this year. They noticed that amounts of collected propolis in winter were 11.79 and 7.30 g / colony for FI and F2, respectively. There were significant differences between seasons during this year.

Hassan (2007) indicated that, the total amount of propolis collected by strong colony was 85.15 g/colony/year and moderate colony was 42.1, while the total amount of propolis collected by weak colony was 10.94 g/colony through the year. While the total amount collected during summer was higher than the amount collected during spring.

Adedoyin et al. (2010) concluded that a total of 456 g of propolis was harvested from two hives in twelve months. in hive 1, the propolis yield in the honey flow period (December– March) was 5 g, 3 g, 5 g and 10 g respectively. They showed that lower than yield in other periods, except the yield in March that was not different from propolis yields in April, May and November. The highest propolis harvest (36 g) took place in August and it was higher than harvest in the other months except in June and July. They found in hive 2, the lowest propolis harvest of 4 g was in December and January. The propolis yield during these months (December–February) was lower than yield in other periods. The highest mean propolis harvest from two hives was in August and it was higher than the harvest in other months except in June and July. A significantly high propolis (74 g) was gathered by the bees from botanical sources during the build up period.

2.2. Biological properties of some bee products :

2.2.1. Honey :

Hakim et al. (2014) assured that, both ginger and Gelam honey have antagonistic effects on colorectal cancer. Joint application of both substances highly the IC50 of Gelam honey to 22mg/ml instead of 75mg/ml when wed alone. Further analysis on the induction of cellular apoptosis indicated that combined treatment of ginger and Gelam honey .resulted in higher apopotosis than 5-FU alone.

Sadeghi‑Aliabadi et al. (2015) revealed the cytotoxic effects of Astragalus honey, ethanol extract of propolis and a sugar solution (as control) against HepG2, 5637 and L929 cell lines evaluated by the MTT assay. Propolis IC50 values were 58, 30 and 15 µg/ml against L929, HepG2 and 5637, respectively. Values for honey were 3.1%, 2.4% and 1.9%, respectively. Propolis extract has increased the expression of the Bcl‑2 gene in all cell lines whereas the honey reduced that significantly (P < 0.05). Honey and propolis reduced p53gene expression in HepG2 and 5637significantly but not in L929 cells. The sugar solution increased the expression of p53 in two cancer cell lines but no significant changes were observed in the expression of this gene in L929 as normal mouse cell.

Tahir et al. (2015) showed that, the IC50 of ginger and Gelam honey alone were 5.2 mg/ml and 80 mg/ml,respectively, whereas the IC50 of the combination treatment was 3 mg/ml of ginger plus 27 mg/ml of Gelam honey witha combination index of < 1, suggesting synergism. Cell death in response to the combined ginger and Gelam honey treatment was associated with the stimulation of early apoptosis accompanied by downregulation of the KRAS, ERK, AKT,Bcl-xL, NFkB (p65) genes in a synergistic manner.Thus, The combination of ginger and Gelam honey may be an effective chemopreventive and therapeutic strategy for inducing the death of colon cancer cells.

Muhammad et al. (2016) concluded that, Acacia honey is of high quality by virtue of the applied melissopalynology. It’s highly nutritional with strong antioxidant and immunomodulatory potentials which may therefore be considered a potential candidate for both cancer prevention and treatment. As a viable therapeutic neurologically, it may be considered agent in the management of Alzheimer’s disease.

2.2.2. Bee venom:

Hamedani et al. (2005) indicated that, the responses of various cell lines against bee venom were different. The increasing amounts of bee venom to human monocyte cell line (K562) revealed a significant increase in proliferative response. The bee venom had no influence on IFN-α production in cell culture media, whereas, adding the BV to K562 cell line could significantly increase the production level of IFN-β only on day 8 post-treatment.

Lee et al., (2007) Showed that, cells were treated with bee venom (BV) at concentrations of 1 or 5 g/ml, BV induced cell death in a time-dependent manner until 24 h, but these cytotoxic effects ended thereafter. They found cells were treated with BV at a concentration of 10 g/ml, however, viability decreased until 72 h, which may have been due to the half-life of BV. Whole BV also inhibited proliferation in these cells. BV induced DNA fragmentation and micronuclei in HL-60 cells and DNA fragmentation in human lymphocytes. Phosphate and tensin homolog (PTEN) up-regulation in HL-60 cells may induce S-phase cell cycle arrest. Forkhead transcription factor (FKHR and FKHRL1) up-regulation in human lymphocytes by whole BV treatment may be involved in the repair of damaged DNA and reduce genotoxicity.

Son et al., (2007) Suggested that, bee venom (BV) has anti-cancer activity. The cell cytotoxic effects through the activation of phospholipase A2 (PLA2) by melittin have been suggested to be the critical mechanism for the anti-cancer activity of BV. They pointed that conjugation of cell lytic peptide (melittin) with hormone receptors and gene therapy carrying melittin can be useful as a novel targeted therapy for some types of cancer, such as prostate and breast cancer.

Ip et al., (2008) showed that, bee venom (BV) induced cell cycle arrest and apoptosis in human breast cancer MCF7 cells. After MCF7 cells were incubated with 10 Ìg/ml BV for 0, 24 and 48 h, they were isolated for examining the effects on cell cycle and apoptosis. BV treatment led to ROS production up to but after treatment led to a decrease in the levels of ROS, which may be associated with the observations of BV affecting glutathion S-transferase (GST), Zn-superoxide dismutase (ZnSOD), Cu/Zn-superoxide dismutase (Cu/Zn-SOD) and catalase. They showed that BV induced DNA damage while DAPI staining also confirmed that BV induced apoptosis in examined MCF7 cells. They found that BV increased the levels of AIF and EndoG in MCF7 cells.

Jeong et al. (2014) showed that, melittin inhibits EGF-induced tumor invasion by suppressing MMP-9 expression and FAK phosphorylation. Mechanistically, the inhibitory effects of melittin on tumor motility and invasion may involve regulating MMP-9 and FAK by suppressing thePI3K/Akt/mTOR signaling pathway, which suggests that melittin is a potential anti-tumor invasion agent for breast cancer therapy. These results can be used for the further study of the development of melittin as a pharmacological agent.

Kohno et al. (2014) investigated the cell entry mechanism of the membrane-lytic peptides K8L9 and melittin in cancer cell lines. K8L9 and melittin interacted with the highly expressed endocytic receptors neuropilin-1, low-density lipoprotein-related protein receptor 1 (LRP1),and transferrin receptor. Silencing of these receptors by small interfering RNAs (siRNAs) reduced the cytotoxic activity of K8L9 in four cancer cell lines.Intracellular K8L9 and melittin triggered enlargement of the lysosomal compartments and cytosolic translocation of cathepsin B. Hsc70 was identified as a melittin-interactive molecule using coimmunoprecipitation and mass spectrometry, and Hsc70-siRNA reduced the cellular uptake of K8L9 and cytotoxic activity by K8L9 and melittin. They suggested that K8L9 and melittin can enter cancer cells via receptor endocytosis following subcytotoxic treatment and subsequently affect lysosomal compartments.

Nittner-Marszalska et al., (2015) showed that, 9a,11b-PGF2 is actively involved in the early allergic response to BV (bee venom) and can be measured during in vivo provocation with BV being potentially a marker useful for monitoringMC (mast cell)activation. They pointed that results of 9a,11b-PGF2 release measurements obtained statistically differentiate BVA (bee venom anaphylaxis) patients from healthy subjects, the method is not promising as a diagnostic instrument due to the results’ high inter-subject variability. They reported that it might be used in the process of monitoring allergic patients, for instance, during VIT (venom immunotherapy). They observed that very interesting is the finding of extremely high response to allergen challenge in some patients, which has not been clinically reflected in the course of immunotherapy. They opined that phenomenon requires further studies conducted in a larger group of patients with their long-term prospective observation.

Zheng et al. (2015) found that, Bee venom (BV) inhibited growth of colon cancer cells through induction of apoptosis. They also found that the expression of death receptor (DR) 4, DR5, p53, p21, Bax, cleaved caspase-3, cleaved caspase-8, and cleaved caspase-9 was increased by BV treatment in a dose dependent manner (0–5 μg/ml). Consistent with cancer cell growth inhibition, the DNA binding activity of nuclear factor kappa B (NF-κB) was also inhibited by BV treatment. Besides, they found that BV blocked NF-κB activation by directly binding to NF-κB p50 subunit. Moreover, combination treatment with BV and p50 siRNA or NF-κB inhibitor augmented BV-induced cell growth inhibition.

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