MELLIFERA 13-26:10-20 (2013) HARUMRESEARCH ARTICLE

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HONEY BEES AND THEIR PRODUCTS AS A BIO-IN-DICATOR OF ENVIRONMENTAL POLLUTION WITH

HEAVY METALSBAL ARILARI VE ÜRÜNLERİNİNİN AĞIR METALLERLE OLUŞAN ÇEVRE KİRLİLİĞİNDE

BİYO-İNDİKATÖR OLARAK KULLANILMASI

Yahya A. Al Naggar1,*, El-Saied A. Naiem2, Amal I. Seif3 and Mohamed H. Mona4

Summary: The aim of this study is to evaluate the effectiveness of honeybees and their associated pro-ducts as biological indicators of the presence of lead, cadmium, copper, iron and zinc in the environment. Samples were collected from four different Egyptian regions with different anthropogenic activities na-mely, Kafr El-Sheikh, El-Mehala El-kobra , Kafr El-Zayat and Al-Fayoum during spring and summer honey harvest 2010. Differences in the concentrations of heavy metals in fresh honey collected during spring and summer honey harvests were observed. Cadmium was found completely absent in fresh honey summer harvest collected from all apiaries. However, the values of Cd in honey samples harvested during spring from the four apiaries exceeded the maximum admitted level (0.05 ppm). In addition, the content of Pb in honey during summer exceeded the maximum admitted level (1.5 ppm) only at Kafr El-zayat. Bee gathered pollen heavy metal contents were higher during spring when compared with those during summer. Cadmium content in pollen collected during spring exceeded the maximum admitted level (0.3 ppm) at all locations except at El-Mehala El-kobra. The reported concentrations of heavy metals decre-ased in the following order: honey bee workers > pollen > honey. These results indicate that honeybees and, to a lesser extent, some of their products (pollen and honey), can be considered as bioindicator of environmental pollution with heavy metals.

Keywords: honey, pollen, bio-indicator, heavy metals, honeybees, environmental pollution.

Özet: Bu çalışmanın amacı, bal arıları ve ilgili ürünlerin kurşun, kadmiyum, bakır, demir ve çinko varlı-ğında oluşan çevre kirliliğinde biyolojik göstergeler olarak etkinliklerini değerlendirmektir.

Bal örnekleri farklı antropojenik aktivitelerin gözlendiği Mısır’ın Kafr El-Şeyh El-Mehala El-kobra, Kafr El-Zayat ve Al-Fayoum adlı dört farklı bölgesinden 2010 yılının ilkbahar ve yaz dönemlerinde hasat edil-miştir. Bahar ve yaz dönemlerinde toplanan taze balların ağır metal konsantrasyonlarında farklılıklar gözlenmiştir. Yazın toplanan taze balların hiçbirisinde kadmiyuma rastlanılmamıştır. Ancak, ilkbaharda hasadı yapılan dört bal örneğinin Cd değerleri maksimum kabul seviyesinden (0.05 ppm) yüksek bu-lunmuştur. Ayrıca, yaz aylarında Kafr El-Zayat bölgesinden alınan balda Pb içeriği maksimum kabul seviyesinden (1.5 ppm) yüksek bulunmuştur. Arıların topladıkları polenlerin ağır metal içerikleri yaz ay-larında bahar aylarındakinden daha yüksek olarak tespit edilmiştir. Bahar döneminde toplanan polenin kadmiyum içeriği El-Mehala El-kobra bölgesi hariç bütün bölgelerde maksimum kabul düzeyininin (0,3 ppm) üstünde bulunmuştur. Ağır metallerin bildirilen konsantrasyonları şu şekilde azalmaktadır: İşçi bal arısı> poleni> bal. Bu sonuçlar, bal arılarının ve daha az ölçüde de olsa ürünlerinin (polen ve bal), bazı ağır metaller aracılığıyla gerçekleşen çevre kirliliğinde biyo-indikatör olarak kabul edilebilir olduğunu göstermektedir.

Anahtar Kelimeler: bal, polen, biyo-gösterge, ağır metaller, bal arıları, çevre kirliliği.

1,2,3,4 Department of Zoology, Faculty of Science, Tanta University 31527, Tanta, Egypt.*Corresponding author E-mail: yehia.elnagar@science.tanta.edu.eg

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IntroductionBiosphere pollution by chemicals and heavy metals ac-celerated dramatically during the last few decades due to mining, smelting, manufacturing, use of agricultural fertilizers, pesticides, municipal wastes, traffic emis-sions, industrial effluents and industrial chemicals etc. Wide occurrence of metal pollution exists worldwide now, including Egypt (Moussa and Abdelkhalek 2007). The extent of environmental pollution and resulting hu-man exposure to hazardous toxic heavy metals in the environment is difficult to assess. Chemical analysis of the environment matrix is the most direct approach to reveal the heavy metal status in the environment, while it cannot afford the powerful evidence in the integrated influence and possible toxicity of such pollution on or-ganisms and ecosystem.

One of the possible alternative approaches to this problem is the use of biological indicators to demon-strate environmental pollution. This approach appears to be particularly suitable for demonstrating exposure to potentially toxic trace elements. Biological moni-toring within a quality control program involves the systematic use of living beings for obtaining quantita-tive information on changes in the environment, often due to anthropogenic activities (Bargagli 1998).Insects provide the cornerstone of biological monitor-ing in aquatic systems, where there are well-developed procedures for using them to assess biological integ-rity. The use of insects as bioindicators in terrestrial ecosystem, in contrast, has been far less enthusiasti-cally embraced.

The honey bee, Apis mellifera L. (Hymenoptera: Api-dae), has been investigated as bio-indicator to moni-tor pollutants (Leita et al. 1996; Bogdanov 2006). Its attractiveness as an ecological detector depends on several features such as high reproductive rate, large flying range since they often forage to 2–3 km away from the apiary, the body is covered with hairs that collect various particles and increase by this mean, close contact with the surrounding environment, sen-sitive to toxic substances, and the possible use of bee products as indicators for environmental pollution (Porrini et al. 2003; Stark 2003). This last point is im-portant because bees and bee products can be used as accumulative and reactive indicators (Billalov et

al. 1992). Apicultural matrix analysis such as honey, wax, pollen or the bees themselves can provide useful indicators of pollution of soil, plant and air pollution by heavy metals in an area of some square kilometers (Porrini et al. 2003; Podgorski and Kanoniuk 2004).

The goal of the present study is to verify the effective-ness of a bio-indicator–based method involving the analysis of honey bee workers Apis mellifera L. (Hy-menoptera; Apidae) and their products (honey and pol-len) for the determination of the environmental pollu-tion with heavy metals by comparing data obtained by different sampling sites in different seasons in Egypt.

Material and MethodsStudy areasThe biomonitoring study was performed in spring and summer during the year 2010 on four sampling sites in Egypt “fig. 1”. Site one, Kafr El Sheikh (A), is lo-cated in the Nile Delta. The apiary selected for this study in Kafr El Sheikh was within agricultural area located near the highway connecting Tanta city and Kafr El Sheikh characterized by motor vehicle circu-lation. The apiary contains 120 bee hives belonging to a beekeeper. The second (B) and the third (C) sites are located in El Mahalla Al Kobra and Kafr El Zyat cities respectively. They represent two major cities in El Gharbia Governorate in the center of the Delta of the Nile. The apiary selected in El Mahalla Al Kobra for this research was located within agricultural field about 500 m away from ElGharbia drainage canal and about 500 m away from El-Mahala-Kafr Elseikh high way. This high way is characterized by intense mo-tor vehicle circulation. The apiary consists of 100 bee hives belonging to a farmer. In kafr El Zyat, the apiary selected for the present study consists of 70 bee hives belonging to a beekeeper. It is situated in agricultural area about 1 km far away from a fertilizer factory. The last one was in El- Fayoum Governorate (D). It is the largest oasis in Egypt which is a depression or basin in the desert to the west of the River Nile in the south of Cairo. The chief crops of the region are cereals, clo-ver, maize and cotton. The research materials for this investigation were obtained from a private beekeeper apiary consists of 150 bee hives inside an agricultural area. The apiary was located 1 km far away from Qa-roune Lake.

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Sampling Honey, stored pollen and honey bee workers were sampled from each bee hive during spring and sum-mer seasons; the routine honey production seasons in Egypt. To avoid contamination, plastic gloves were used in sampling and handling honey and pollen.

HoneyIn Egypt, there are several types of honey. For each type there is four stages of maturation; freshly col-lected, sealed, extracted and ripened honey. In order to avoid any kind of metal contamination, fresh honey samples harvested from bee hives during spring and summer were used in this work. Fresh honey was squeezed out from the honey comb into a small dis-posable polyethylene container.

PollenPollen samples were collected by cutting a piece of comb (6 cm2) containing stored pollen using dispos-able plastic knife.

Honey bee workersHoney bee workers were carefully brushed directly into a disposable polyethylene bags after fuming by smoker to calm the bees. The bees were collected from the honey combs located on the farthest side walls of the hive. These were normally old bees probably more exposed to airborne pollution. After the collecting trip, all samples were frozen at - 4 oC in the laboratory (Fakhimzadeh and Lodenius 2000).

Samples preparationFresh honey samples were used directly without any kind of preparation. The pollen and bees were dried overnight at 105 oC on drying oven then finely ground by grinder and packed in plastic bags and stored for heavy metal analysis (Fakhimzadeh and Lodenius 2000).

Heavy metal analysis In this series of experiments, quantitative determina-tion of heavy metals in honey bee workers and bee products was performed by flame atomic absorption

Figure 1. Satellite view showing sampling sites A, B, C and D (Google Earth Gold Pro).

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spectrophotometer (FAAS). A Perkin-Elmer Model 2380 atomic absorption spectrophotometer (USA) was used. The instrumental conditions for each ele-ment were taken from the instrumental manual (Price, 1979; Welz and Sperling 1999).

The readings were taken from the device and were converted to actual concentration of metals in the samples using the equation;Heavy metal conc. (ppm) = R. conc. D. fWhere;R. conc. = Reading concentration, S. v = Sample vol-ume, S. wt = sample weightand D. f = Dilution factor.

Analytical procedure for heavy metals in honey bees and their productsHoneySamples were prepared according to 920,180 method of the AOAC (2000). Five mL of 75 (w/v) % HNO3 were added to each 2 g sample of honey within a por-celain crucible. The acid was then evaporated on an electrically heated hot plate at 100-120 ºC, and then it was gauged to 10 mL with distilled water (dilution 1:5). Finally, the sample solution was analyzed for their heavy metal concentrations by FAAS technique. PollenPollen samples were prepared according to method described by Nuorteva (1990). About 0.5 g of each finely ground and dried pollen sample was weighed and digested in 5 mL conc. HNO3 for 1 h at 75 ºC and 4 h at 105 ºC. Wet ash was filtered and diluted with distilled water to 25 mL. The sample solution was analyzed for its heavy metal concentrations by FAAS.

Honey bee workersHoney bee samples were prepared according to meth-od described by Nuorteva (1990). The ground dried bee samples were weighed to ≤ 0.5 g and dissolved into 5 mL HNO3 for 2h at 65 ºC, then 5 mL of H2O2

were added, and the samples were heated for addition-al 2h at 65 ºC. The samples were filtered and diluted with distilled water to 25 mL. The sample solutions were analyzed for their heavy metal concentrations by FAAS.

Maximum admitted levels of heavy metalsThe maximum admitted levels (MAL) of Cu, Zn, Cd, Pb and Fe in honey, bee gathered pollen and ani-mal tissues were compared according to Allen et al. (1974), Kabata-Pendias (1984, 1992), The German Federal Ministry of the Environment (1992), Kamran (2000), Piven et al. (2003), Luliana and Cecilia (2005) and Tuzen and Soylak (2005).

Results and DiscussionHoneyThe mean concentrations of Cu, Zn, Cd, Pb and Fe in honey samples obtained from bee hives in Al-fayoum, Kafr El-zayat, El-mehala El-kobra and Kafr El-sheikh are given in “tab. 1” and the values were ranged from 0.00 to 11.20 ppm in the samples.

According to the obtained data, differences in the concentrations of heavy metals in the fresh honey col-lected during spring and summer harvests at the four different locations were observed. The highest mean concentrations of Zn (1.43 ppm), and Pb (1.59 ppm) were recorded in honey harvested during summer from apiary in Kafr El-zayat. The highest level of Fe was observed in honey collected during spring from Al-Fayoum bee colony. Meanwhile, the maximum mean concentration of Cd (0.14 ppm) was in spring honey harvested from apiaries in both El-Mehala El-kobra and Al- Fayoum.

Interestingly, Cd was completely absent in fresh hon-ey summer harvest collected from apiaries in Kafr El-zayat, Kafr El-sheikh, El-mehala El-kobra and Al- fayoum. At these areas, honey was mostly produced by honey bees from nectar of cotton flowers. A simi-lar trend was observed for Zn. It was absent in spring honey harvest collected from apiaries in Al-fayoum and El-mehala El-kobra. Honey was produced by bees in these sites to a large extent from clover blossom nectar. The present study links pollution of area with honey. The contamination sources can be roughly divided into environmental and apicultural ones. In areas lo-cated near industrial areas and, indeed, the level of Pb in honey was higher than isolated areas like oasis Al-Fayoum. Therefore, because of the high risks of

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contamination the apiaries should be located in areas situated at long enough distance from any possible pollutant source and not near the pollutant areas. The results reported herein, indicated that the concen-trations of Cu, Cd and Fe in honey samples collect-ed during spring were higher than those reported in honey samples collected during summer. These find-ings were disagree with those of El-Sherbiny and Rizk (1979) who stated that, cotton honey during summer contained higher element concentration than clover honey during spring .These differences may be attrib-uted to the floral type of honey plants, floral density and composition of nectar and pollen (Abu- Tarboush et al. 1993; Torres et al. 2005; Silici et al. 2008). Since, during spring the main source for nectar and pollen was clover blossoms. Meanwhile, the main sources for nectar and pollen were cotton, maize and pumpkins during summer.

The concentrations of heavy metals in honey have been detected in several studies. Comparison between the present data and previous studies it was found that, the concentrations of heavy metals in honey samples were higher than those reported by Hassan et al. (2010) except for Pb since; they recorded a slightly higher level of Pb (0.007- 1.65 ppm). Similarly, heavy met-als concentrations in honey samples were higher than those reported by Staniškienė et al. (2006) and Tuzen and Soylak ( 2005) except for Zn since; they recorded a significantly higher level of Zn (1.1- 24.2 ppm). On the other hand, data reported herein for heavy met-als content in honey were lower than those reported by Belouali et al. (2008). These differences may be attributed to the different geographical and botanical sources of these metals in honey.

All values recorded for Cd and Fe in fresh honey sam-ples harvested during spring were higher than the MAL (0.05 ppm and 5.2 ppm) reported by Piven et al. (2003) and Tuzen and Soylak (2005) except of iron in honey samples from El-Mahala El Kobra apiary. Mean while, the concentrations of Pb exceeded MAL (1.5 ppm) reported by Piven et al. (2003) in honey samples harvested during summer from Kafr- el -zayat apiary only. These findings confirmed the elevated levels of environmental pollution with these metals and at the same time emphasized the possibility of using honey as

a bioindicator for environmental pollution with heavy metals (Porrini et al. 2003; Podgorski and Kanoniuk 2004; Tuzen and Soylak 2005; Bogdanov 2006).

PollenThe contents of heavy metals in honey bee gathered pollen collected from apiaries in agricultural areas were determined “tab. 2” Data obtained revealed that, the highest levels of Cu (24.18 ppm), Zn (42.42 ppm) and Fe (95.6 ppm) were noticed in bee pollen collected during spring from apiary in Kafr El-sheikh. Mean-while, the maximum levels of Cd (1.38 ppm) and Pb (14.23 ppm) were reported also during spring but in bee pollen collected from apiaries in Al-fayoum and El-mehala El-kobra respectively. On the other hand, the lower concentrations of Zn (0.18 ppm), Cd (0.03 ppm) and Pb (0.65 ppm) were recorded in pollen col-lected from Al-fayoum apiary. In general, heavy metal concentrations in pollen samples have their maximum level during spring; the season of clover cultivation.

The differences in concentrations of heavy metals in bee pollen between the four study apiaries could be attributed to different local contamination/pollution sources. The main contaminants are heavy metals (Szczesna et al. 1993; Jablonski et al. 1995; Leita 1996; Conti and Botre 2001), pesticides originating from the environment (Fieche et al. 1997; Kubik et al. 1999) and from agricultural and apiculture practices (Zhou et al. 1994).

Surprisingly, high contamination of cadmium in pollen collected from apiary in Al-fayoum was detected. This may be due to the use of excess amount of cheap fer-tilizers and pesticides containing cadmium. Since, the phosphate fertilizers are generally the major source of trace metals among all inorganic fertilizers and much attention had been paid to the concentration of Cd in phosphate fertilizers (Lu et al. 1992). Roman (2009) reported that, pollen collected from the agricultural woodland in Poland had high concentration of cadmi-um. Similarly Morgano et al. (2010) reported elevated concentrations of Pb and Cd in the pollen in Brazil.

There are very few published studies on heavy metal contamination in bee gathered pollen. Migula and Kafel (1993) reported that Cd was present in pollen

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Table I. Heavy metal concentrations (ppm) (Mean ± SD) in honey samples collected from bee hives at El-Mehala El-kobra, Kafr El-Sheikh Kafr El-Zayat, and Al- Fayoum during spring and summer (2010).

Season Location

Element conc.(ppm)

Copper (n=3)

Zinc (n=3)

Cadmium (n=3)

Lead (n=3)

Iron (n=3)

Spring

El-Mehala El-kobra 1.65±0.07 0.00±0.00 0.14±0.04 0.35±0.05 3.53±2.67

Kafr El-Sheikh 3.23±3.23 0.05±0.05 0.13±0.13 0.32±0.32 6.56±0.55

Kafr El-Zayat 1.47±0.04 0.28±0.01 0.07±0.01 0.26±0.13 7.10±0.05

Al-Fayoum 2.37±0.05 ND 0.14±0.00 0.08±0.01 11.20±0.05

Summer

El-Mehala El-kobra 0.14±0.06 1.16±0.05 ND 0.11±0.03 3.73±0.16

Kafr El-Sheikh 0.12±0.04 1.32±0.16 ND 0.68±0.12 2.80±0.13

Kafr El-Zayat 0.15±0.05 1.43±0.05 ND 1.59±0.46 3.58±0.30

Al- Fayoum 0.11±0.01 1.02±0.03 ND 0.87±0.18 3.03±0.10

ND* (not detected)

Table 2. Heavy metal concentrations (ppm) (Mean ± SD) in bee pollen collected from bee hives at El-Mehala El-kobra, Kafr El-Sheikh Kafr El-Zayat, and Al- Fayoum during spring and summer (2010).

Season LocationElement conc. (ppm)

Copper( n = 3)

Zinc(n = 3)

Cadmium(n = 3)

Lead(n = 3)

Iron(n = 3)

Spring

El-Mehala El-kobra 6.07 ± 0.93 7.13±0.48 0.18 ± 0.28 14.23 ± 4.33 93.83 ± 2.08

Kafr El-Sheikh 24.18 ± 0.44 42.42 ± 0.95 0.75 ± 0.25 4.75 ± 0.10 95.6 ± 10.13

Kafr El-Zayat 22.28 ± 4.54 20.25 ± 0.31 0.82 ± 0.10 4.80 ± 0.10 85.68 ± 0.76

Al-Fayoum 11.53 ± 0.75 0.18 ± 0.28 1.38 ± 0.26 0.65 ± 0.10 43.33 ± 1.26

Summer

El-Mehala El-kobra 8.13 ± 2.71 19.21 ± 0.63 0.067 ± 0.03 10.53 ± 0.49 30.83 ±3.9

Kafr El-Sheikh 1.90 ± 0.41 21.87 ± 1.18 0.10 ± 0.05 13.85 ± 1.63 20.33 ± 1.53

Kafr El-Zayat 0.95 ± 0.18 20.40 ± 4.16 0.08 ± 0.03 10.13 ± 9.77 31.83 ± 5.03

Al-Fayoum 7.62 ± 0.61 31.30 ± 0.35 0.03 ± 0.06 2.85 ± 0.79 24.00 ± 1.32

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loads collected in the apiary near road. Konopacka et al. (1993) reported that in as many as 50% of pollen samples,

In the current study, all the highest mean concentra-tions of heavy metals recorded in the pollen were dur-ing spring. These differences may be related to the bo-tanical source of pollen in each season since, in spring the main source of pollen is clover while in summer the pollen may be collected from many plants as cot-ton, maize, eucalyptus and pumpkins. In the present study, Pb levels in pollen collected from beehives in El-mehala El-kobra, Kafr El-sheikh and Kafr- El-zyat considerably exceeded permissible standard (3 ppm), but the content of Pb in pollen was below permissi-ble standard in Al-fayoum .Mean while, Cd levels in pollen collected during spring from beehives in Kafr El-sheikh, Kafr- El-zyat and Al-fayoum were higher than those of the maximum admitted levels (MAL) (0.3 ppm) reported by Allen et al. (1974).These observations as previously mentioned emphasize the elevated level of environmental pollution with these toxic metals.

Honey bee workersThe present study revealed that the highest mean val-ues of Cu (29.17 ppm) and Pb (16.67 ppm) were re-ported in worker honey bees during winter at Kafr El-sheikh. Meanwhile, the maximum mean concen-trations of Cd (3.20 ppm) and Fe (447.17 ppm) were noticed also during winter but in honey bees collected from apiary in Al-fayoum. On the other hand, the high-est mean concentration of Zn (77.95 ppm) in bees was recorded during spring at kafr El-Zayat “Tab. 3”. The results of the present study reinforce the idea that high concentration of heavy metals reflected in the honey bee worker due to local environmental exposure. Several investigators commend upon the possibility to use bees to monitor the purity of the environment (Crane 1984; Muszynska 1995; Zarski et al. 1996; Ro-man 1997, 2000,). Birge and Price (2001) carried out a study to determine the heavy metals concentrations in honey bee workers. The concentrations of heavy metals ranged between 3.53- 6.26, 27.65- 30.80, 0.05- 0.19, 375.4- 446.5 and 3406.35- 5161.25 ppm for Cu, Zn, Cd, Pb and Fe respectively. Similarly, Conti and Botre (2000) found the concentrations of Cd and Pb

in honey bee workers ranged between 2.87- 4.23 ppm for Cd and 0.61- 1.25 ppm for Pb. The concentrations of Cu, Zn and Cd reported herein were higher than those reported by Birge and Price (2001) and Pb was higher than those reported by Conti and Botre (2000). These differences may be attributed to the varying de-grees of heavy metals contaminations at each location (Zhou et al. 1994).

According to the results obtained in the present study, some of the recorded values for Cd, Pb and Fe in honey bee workers collected from aparies at El-mehala El-kobra, Kafr El-sheikh, Kafr El-zayat and Al- fayoum during spring and summer (2010) were higher than MAL) Cd (0.5 ppm), Pb (3 ppm) and Fe (400 ppm) reported by Allen et al. (1974).This finding further emphasize the elevated level of environmental pollution with these metals at such locations and the possibility of using honey bee workers as a bioindica-tor for environmental pollution with heavy metals.

Comparison of heavy metals conc. (ppm) in honey, pollen and honey bee workers “Fig. 2” showed a comparison between the total means of heavy metal concentrations (ppm) of honey, pollen and honey bee workers. Heavy metal concentrations (ppm) took the following order: honey bee workers > pollen > honey. These findings were in a good agreement with Höf-fel (1982) and Altmann (1983) who reported that, the degree of contamination with heavy metals decreased in the following order: bees ≥ propolis > comb wax > honey.

Relationship between honey, pollen and honey bee workers heavy metals concentrations (ppm) as envi-ronmental variables The most effective environmental variable of heavy metals contents of honey, pollen and bees samples were indicated on the ordination diagram produced by principal component analysis (PCA). Since, CANOCO ordination axes represent the relationship between the heavy metals concentrations variables of honey, pol-len and honey bee workers “Fig. 3A-C”. Iron was the most effective environmental variable of heavy metal contents followed by Zn, Cd, and Cu in fresh honey samples collected during spring and summer harvests

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at the four different apiaries. In contrast, Pb was the least effective environmental variable of heavy metal contents of fresh honey samples “fig. 3A”.

Mean while, lead was the most effective environment variable of pollen heavy metals contents than Cd, Cu and Fe. In contrast, Zn was the least environmental variable of the pollen heavy metals concentrations

“fig. 3B”. On the other hand, Iron was the most effec-tive environmental variable in the honey bee workers heavy metal concentrations then Cd, Cu and Pb. On the other hand, Zn was the least environmental vari-able in the honey bee’s heavy metals concentrations collected from aparies at El-mehala El-kobra, Kafr El-sheikh, Kafr El-zayat and Al- fayoum during spring and summer (2010) “fig. 3C”.

Table 3. Heavy metal concentrations (ppm) (Mean ± SD) in honey bee workers collected from aparies at El-Mehala El-kobra, Kafr El-Sheikh Kafr El-Zayat, and Al-Fayoum during spring and summer (2010).

Season LocationElement conc. (ppm)

Copper( n= 3)

Zinc( n= 3)

Cadmium( n= 3)

Lead ( n = 3)

Iron ( n= 3)

Spring

El-Mehala El-kobra 10.85±0.75 72.17±4.04 0.25±0.20 11.15±8.46 201.17±8.14

Kafr El-Sheikh 13.83±3.23 13.80±2.99 1.60±0.10 2.32±0.24 134.00±2.78

Kafr El-Zayat 17.22±0.56 77.95±1.87 0.92±0.23 10.58±4.50 336.33±13.7

Al-Fayoum 18.17±1.95 35.63±0.76 0.72±0.21 8.18±10.67 225.33±16.0

Summer

El-Mehala El-kobra 3.62±0.79 46.57±0.48 0.10±0.05 5.67±0.76 73.00±1.80

Kafr El-Sheikh 6.62±1.33 44.25±2.86 0.10±0.05 8.50±0.50 74.00±2.29

Kafr El-Zayat 7.77±0.65 46.42±8.28 0.07±0.03 11.23±6.52 77.50±8.50

Al-Fayoum 7.50±1.78 67.17±7.14 0.15±0.05 2.95±2.13 147.00±18.6

0102030405060708090

100110120130140150160170180

Fe

Con

c. (

ppm

)

Honey Pollen Honey bee workers 048

1216202428323640444852

cu zn cd pb

Conc

. (pp

m)

Figure 2. Comparison of heavy metals contents (ppm) in honey, pollen and honey bee workers.

MELLIFERA 18

ConclusionData presented in this work are part of a study aimed to verify whether honeybees and their products can be considered reliable biological matrices for the as-sessment of heavy metal pollution. Specifically, the present work designed to verify whether it could be possible to detect significant variations in the values of heavy metals concentration among samples col-lected from sampling sites characterized by markedly different environmental impacts. For this reason, the collection and analysis of samples refer to freshly pro-duced materials. Finally, our results indicated that the contents of heavy metals in some collected samples of honey, pollen and honey bee workers exceeded the

maximum admitted levels of these metals. Therefore, these findings emphasize the elevated level of envi-ronmental pollution with these metals at such loca-tions and the possibility of using honey bee workers and to a lesser extent, some of their products (pollen and honey) as a bio-indicator for environmental pollu-tion with heavy metals.

AcknowledgmentWe thank all the apiarists who participated in the pre-sent study for their time. Heavy metals analysis has been done at flame atomic absorption spectrophotom-eter (FAAS) unit, Central laboratory, Tanta Univer-sity, Egypt, which is gratefully acknowledged.

Figure 3. CANOCO ordination plot indicating the relationship between heavy metals concentrations (ppm) as environ-mental variables of Honey (A), Pollen (B) and honey bee workers (C) collected during spring and summer 2010 at the four different apiaries.

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