Origin and sources of Aliphatic and Polycyclic

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Origin and sources of Aliphatic and Polycyclic aromatic. Hydrocarbons in Water, Sediment and Fish of Lake Burullus,. Egyptian Mediterranean Sea. Tarek O.
Origin and sources of Aliphatic and Polycyclic aromatic Hydrocarbons in Water, Sediment and Fish of Lake Burullus, Egyptian Mediterranean Sea Tarek O. Said* National Institute of Oceanography and Fisheries, Kayet Bay, Alexandria-Egypt. [email protected] Fax: +2 03 4801174 Handy Phone: +2 0105047184

Abstract Lake Burullus is one of the Delta lakes, connected with the Mediterranean Sea through El Boughaz opening. The compositions of PAH determined in the dissolved fraction of seawater were measured in order to use them as chemical markers for identifying different sources of pollution in the Lake. PAHs were determined in fish tissues for comparison with human health standards as consumption. ∑ALIP varied from, 289-4538, 99-574, 13-3000 and 29-7317µg/l, during winter, spring, summer and autumn, respectively. The sum of ∑PAHs concentrations varied from, 0.07-1.27, 1.01-42.72, 0.5-68.01 and 0.47-9.99µg/l during winter, spring, summer and autumn, respectively. The most dominant PAH fraction was InP with an annual average value of 10.04 µg/l. The calculated Phe/An ratio of 0.53 of Lake Burullus for all samples during the four seasons indicated that the major PAH input was from combustion of fossil fuel via pyrolytic process. The ∑COMB concentrations displayed values from 0.14 to 119.24 µg/l, representing average 70% of total anthropogenic PAHs. ∑PAH CARC ranged from 0.02 to 7.14 with an average value of 3.04 µg/l and an average of 13.28% of total PAHs. The average annual concentration of ∑THC was 959.8µg/l, resulted from all sources into Lake Burullus. The concentrations of individual PAH recorded in sediment of the present study ranged from non-detectable levels to 17556 ng/g, dry weight and were much lower than the ERM values. High concentration of DBA (above the ERM value) at some locations of the Lake might need a more detailed study. BaP recorded in all fish species of the investigated area exceeded a limit of 10ng/g. In addition, contamination by PAHs differs according to kind of fish species, where Clarries sp collected from the Lake have higher ∑PAHs than that recorded in Oresochromus sp. Keywords: PAHs; origin; Lake Burullus; Mediterranean; Egypt. 1. Introduction Estuaries and coastal areas have traditionally, been sites of industrial activity because of ease of transport and because the sea offers a convenient place for the disposal of waste substances. * corresponding author E-mail: [email protected]

GESAMP [1] stated that at least 40% of all marine contaminants are coming from land-based sources. Mediterranean Sea appears to suffer from high anthropogenic pressure due to inputs from; industrial, sewage effluents, storm water drains, shipping activities, spillage, rivers, atmospheric-fallout, coastal activities and natural oil seeps [2]. The annual inputs of petroleum hydrocarbons are about 750 × 103 tons, among which the land-based industrial inputs amount are about 221 × 103 tons per year [3]. Lake Burullus is the second largest natural lake in Egypt with respect to area and production. It is the main source of fish production in Kafr El-Sheikh Governorate, Egypt. Fish catch from the lake has been dropped from about 52.5 thousands tons in 1990 to about 43.6 thousands tons in 1993 or dropped by about 17% [4]. Untreated domestic wastewaters with agricultural and industrial wastes are still released through a number of drainages and outfalls along the coastal area of the Lake. Huge amounts of drainage water enter the lake from several drains with about 2.46x109 m3y-1 of brackish water through different drains: 1) El Burullus drain at the eastern side of the lake; (2) drains 7,8,9 and 11 (Figure 1) in the southern side of the lake; (3) Brembal canal in the western extremity of the lake receives fresh water directly from Rosetta branch during the flood periods; (4) western Burullus and (5) El Nasser and El Gharbia drains. The lake receives also about 100x106m3 y-1 of precipitation, compared with the present size of the lake, the residence time of water takes about 2.5 months [5]. The existing environmental problems of Lake Burullus are mainly related to natural and man-made influences. The direct effect of the increased wind speed on the lake is manifested by the introduction of the seawater into the lake through ElBoughaz channel by the northern wind, the magnitude of which depends on the wind velocity and duration. According to the shallowness of the lake, the increased wind velocity causes also a turbulence of water by stirring up surface sediments. In addition, the wind movements play an important role in the distribution of salinity in the lake. When it is easterly winds, drains of freshwater covers most of the lake and decreases the salinity largely. The northerly winds drive water southerly and the salinity increases even next to drains. Polycyclic aromatic hydrocarbons (PAHs) are a group of about 10,000 compounds, a few of which occur in considerable amounts in the environment. PAHs comprise fused aromatic rings and do not contain heteroatoms or carry substituents. PAHs containing up to four fused benzene rings known as light PAHs and those containing more than four benzene rings known as heavy PAHs. Heavy PAHs are more stable and more toxic than light ones. PAHs are lipophilic in nature; nevertheless, some of them can dissolve quite well in water [6]. Most PAHs in the environment derive from incomplete burning of carbon containing materials, such as oil, wood, garbage or coal. A maximum amount of PAHs is formed when materials burn at temperatures in the range 500–700 °C, as in wood fires or cigarettes. PAHs from fires can bind to ashes and move long distances through the air as some PAHs (especially the lighter ones) are water-soluble, they can also be found in rivers and groundwater [7]. PAHs are among the most carcinogenic and toxic contaminants found in aquatic systems [8]. There is much evidence that PAHs are responsible for massive deaths for habitats [9]. PAHs occur in crude oil, and formed during burning of oil have adverse health effect. They cause irritation to eyes and skin, cancer, possible reproductive effects, immune systems effects [10]. Therefore, monitoring of

such contaminants must be performed using selected bio-indicator species whose biology, biochemistry and contaminant uptake characteristics are well known. This paper aimed to study the distribution aliphatic and polycyclic aromatic hydrocarbons fractions in water, sediment and two types of the most famous fish muscles in Lake Burullus. The effect of different drains water on the distribution of different hydrocarbons pollutants was studied. In addition, calculating the annual inputs of pollutants from the lake into the Mediterranean Sea. In addition, this work aimed to give a brief overview on the current legislative situation of PAH analysis in the environment and to highlight the needs for further investigations and research, especially PAH-monitoring databases as basis for further risk assessments. 2. Materials and methods 2.1 Area of study Lake Burullus is a shallow slightly brackish water situated along the Mediterranean Sea coast. It is one of the Nile delta lakes located between the two main delta promontories; Rosetta and Damietta. It lies on the eastern side of Rosetta branch of the River Nile, Egypt and occupies a central position along the Mediterranean coast of the Nile. It lies between Longitude 30o 30- and 31o 10- E and latitude 31o 21- and 31o 35- N. It has an irregular elongated shape and connected to the sea through a narrow passage (50 m width) called Al-Burg inlet or Boughaz Al-Burullus (figure 1). The present area of lake Burullus is about 420 km2 (100000 feddan) of which 370 km2 is open water. Former estimates of the area are 588 km2 (140000 feddan) in 1956 and 462 km2 (110000 feddan) in 1974 [5]. It seems that during the last 10 years there has been a reduction in the lake area by 30%. This decrease is due to continuous land reclamation projects along the southern and eastern shores of the lake and fish farming processes. The length of the lake is about 47 km, and the width varied between 6 and 16 km, with an average of about 11 km. The depth of the lake ranges between 0.42 and 2.07m. 2.2 Sampling activities Twelve surface water samples (1m depth) were collected seasonally from Lake Burullus on R/V Salsabil using a Nisken bottle during 2006. The studied area were represented by four regions inside the lake: Eastern region (stations 4, 5 and 6), western region (station 8), northern region (station 7) and southern region (2 and 3). In addition to stations 9, 10, 11 and 12 lie outside the lake (figure 1). Surface sediment (0-3 cm) samples were collected using a stainless steel grab sampler from 12 sites during August 2006 and stored in pre-cleaned aluminum containers and frozen in a deep freezer at−20°C until analysis. In addition, two types of fish species; Oresochromus niloticus and Clarries sp., were collected during the same period to study the accumulation of organic pollutants. The samples were analysed for aliphatic and aromatic hydrocarbons following, well established technique [11]. 2.2 Extraction 2.2.1 water Seawater samples were extracted three times with 60 ml of dichloromethane in a separating funnel. Sample extracts were combined and concentrated by rotary

evaporation to 5 ml. Finally, samples were concentrated under a gentle stream of pure nitrogen to a final volume of 1 ml [12].

Figure 1. Lake Burullus showing the sampling stations. 2.2.2 Sediment The sediments were freeze-dried, dry/wet ratios determined and then sieved through a stainless steel mesh (250 µm). Each sediment sample (30 g) was Soxhlet extracted with 250 ml of hexane for 8 hours and then re-extracted for 8 hours into 250 ml of dichloromethane [13]. Then the extracts were combined and concentrated down using rotary evaporation at 30°C followed by concentration with nitrogen gas stream down to a volume 1 ml. 2.2.3 Fish Fish tissue (10 g of wet weight of muscles) was placed in a blender and 30 g anhydrous sodium sulfate was added. They were manually homogenized to determine whether the samples were adequately dried. Samples were blended at high speed until the mixture was well-homogenized (2-3 min). The mixture was transferred to a precleaned extraction thimble and the dehydrated tissue was extracted with 200 ml (1:1) n-hexane-dichloromethane for 8hr in a soxhelt apparatus cycling 5 to 6 times per hr, anhydrous sodium sulfate (30 g) was extracted in the same fashion as the sample and used as the blank. The extracted solvents were concentrated with a rotary evaporator down to 2ml (maximum temperature: 35°C), followed by concentration with pure nitrogen gas stream down to a volume of 1ml. Clean-up and fractionation was performed by passing the extract through a silica/alumina column. The extracted volume (water, sediment and fish) was passed through the silica column prepared by slurry packing 20 ml (10 g) of silica, followed by 10 ml (10 g) of alumina and finally 1g of anhydrous sodium sulphate. Elution was

performed using 40 ml of hexane (aliphatic fractions), then 40 ml of hexane/dichloromethane (90:10) followed by 20 ml of hexane/dichloromethane (50:50) (which combined contain PAHs). Finally, eluted samples were concentrated under a gentle stream of purified nitrogen to about 0.2 ml, prior to be injected into GC/FID for different hydrocarbon fractions analysis [11]. 2.4 Gas chromatography All concentrated samples were analysed by a Hewlett Packard 5890 series II GC gas chromatograph equipped with a flame ionization detector (FID). The instrument was operated in split less mode (3μl split less injection) with the injection port maintained at 290 °C and the detector maintained at 300 °C. Samples were analysed on a fused silica capillary column HP-1, with 100% dimethyl polysiloxane (30m length, 0.32mm i.d., 0.17μm film thickness). The oven temperature was programmed from 60 to 290°C, changing at a rate of 3 °C min−1 and maintained at 290 °C for 25 min. The carrier gas was nitrogen flowing at 1.2 ml min−1. 2.5 Quantification A standard mixture of aliphatic fractions (50 mgl-1) contains C10-C30, in addition to pristine and phytane compounds using chloroflourobenzene and C40 as internal standards. A stock solution containing the following PAHs (10 mgl-1) was used for quantification: naphthalene (Naph), acenaphthylene (Acthy), acenaphthene(Ace), fluorine (Fl), phenanthrene (Phe), anthracene (An), fluoranthene (Flu), benzo(a)anthracene (BaA), chrysene (Chr), benzo(b)fluoranthene (BbF), benzo(k)fluoranthene (BkF), pyrene (Py), benzo(a)pyrene (BaP, dibenzo(a,h)anthracene (DBA), benzo(ghi)perylene (BghiP), and indeno(1,2,3-cd) pyrene (InP) by dilution to create a series of calibration standards of PAHs at 0.1, 0.25, 0.5, 0.75, 1.0, 2.0, 5.0, and 10 μgml−1. The detection limit was approximately 0.01μgml−1 for each PAH. For analytical reliability and recovery efficiency of the results, six analyses were conducted on PAH reference materials, HS-5 and 2974 (provided by EIMP-IAEA).The laboratory results showed a recovery efficiency ranging from 91 to 110% with a coefficient of variation (CV) of 9–14% for all studied pollutants (16 PAHs fractions). All solvents were of pesticide grade purchased from Merck, and appropriate blanks (1000-fold concentrates; two for each batch of analysis for both of water and fish samples) were analysed. 3. Results and discussion 3.1 Water Tables 1-4 present the total concentrations of the aliphatic hydrocarbons ranging from C10 to C30, as well as the isoprenoid hydrocarbon individual concentrations (pristine and phytane; Pr and Ph). The concentrations below their limits of detection were given a value of zero for the calculation. In the present study, the total aliphatic hydrocarbon concentrations (∑ALIP) were varied from, 289-4538, 99-574, 13-3000 and 29-7317µg/l, during winter, spring, summer and autumn, respectively. Higher concentrations occurred at station 12 averaged of 1916 µg/l, while lower concentrations occurred at station two averaged of 196 µg/l (Figure 2).

The isoprenoid hydrocarbons, pristane (Pr) (2,6,10,14-tetramethyl pentadecane) and phytane (Ph) (2,6,10,14-tetramethyl hexadecane), are products of geologic alteration of phytol and other isoprenoidyl natural products, and are not primary constituents of most terrestrial biota [14,15]. 25

2500 PAHs 2000

ALIP

15

1500

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1000

5

500

0

0

1

2

3

4

5

6

7

8

AILP (µg/l)

PAHs (µg/l)

20

9 10 11 12

Stations

Figure 2 Average distribution of ∑ALIP and ∑PAH in water of Lake Burullus during 2006. Pristane and phytane are presented in most petroleum oils in a ratio of pristine/phytane (Pr/Ph) lower than 1, so the detection of these two components is often used as good indicators of petroleum contamination. However, a high concentration of pristane alone can be derived from zooplankton [16]. Most samples had Pr/Ph ratios lower than 1 (Tables 1-4) indicating mainly a petrogenic hydrocarbon inputs to Lake Burullus. Highest ratio of Pr/Ph was recorded at stations 1,6,8 and 9, reflecting biogenic origins. The residues of 16 PAHs were identified in water (Tables 1-4). The total (∑16 PAHs) concentrations were varied from, 0.07-1.27, 1.01-42.72, 0.5-68.01 and 0.479.99µg/l during winter, spring, summer and autumn, respectively with an annual average of 43.93µg/l (Table 5). The maximum concentration of ∑PAHs was record ed at stations 9 and 11 while, the minimum ∑PAHs was recorded at station 10 (Figure 2). The most dominant PAH fraction was InP with an annual average value of 10.04 µg/l (Table 5). Total ∑16 PAHs recorded in Lake Burullus ranged from 0.53 to 153.24 µg/l with an average of 43.93µg/l are higher than that recorded in the Red Sea water ranged from 0.4-96.45µg/l with an average of 20.93µg/l [17], and that recorded in Suez Canal ranged from 0.28-39.57µg/l with an average of 10.78 µg/l [18]. Ratio values such as phenanthrene/anthracene (Phe/An) and fluoranthene/pyrene (Flu/Py) had been used by different workers to identify the origin of hydrocarbons [19,20,21]. Petroleum often contains more phenanthrene relative to anthracene as phenanthrene that is more a thermodynamically stable tricyclic aromatic isomer than anthracene, so a Phe/An ratio is observed to be very high in PAH petrogenic pollution, but low ratio in pyrolytic contamination cases. The calculated Phe/An ratio of 0.53 (Table 5) of Lake Burullus for all samples during the four

seasons indicated that the major PAH input was from combustion of fossil fuel via pyrolytic process. In addition, the calculated Flu/Py ratio 2.19 suggested that the origin of PAH was related to a pyrolytic origin. This is in accordance with Garrigues et al. [22] and Benlahcen et al. [23] who stated that Flu/Py ratio less than 1 suggested that the major PAH was from petrogenic inputs, and values greater than 1 were related to pyrolytic origin. The BaA/Chr ratio has also been suggested to identify PAH sources, and this ratio tended to increase as petrogenic contribution decreased. The ratio values for crude and fuel oil ranged from 0.24 to 0.4 [13, 24]. The BaA/CHR ratio in the present study was 2.54 (Table 5); suggesting that the main source of PAH contamination in Lake Burullus came from crude or fuel oil. The IARC probable and possible human carcinogens∑PAH CARC was ranged from 0.02 to 7.14 with an average value of 3.04 µg/l and an average % of 13.28 of total PAHs (Table 5). On the other hand, the low MW < 178 PAH includes Naph, Acthy, Ace and Fl can be derives mainly from fossil sources rather than from microbial activity. In this study, the ∑FPAH (Table 5) varied from 0.39 to 33.99 µg/l with an average of 9.33 µg/l, accounting for an average of 30% of the total anthropogenic PAHs. Contributions from combustion/pyrolysis of fossil fuel can be assessed by considering un-substituted PAH with MW > 178 includes Phe, An, Flu, Py, BaA, Chr, BbF, BkF, BaP, BghiP, DBA and InP. The ∑COMB concentrations displayed values from 0.14 to 119.24 µg/l, representing average 70% of total anthropogenic PAHs (Table 5). The Sum of five carcinogenic PAHs (∑PAH CARC ; sum of BaA+BbF+BaP+DBA+InP) recommended by IARC [25] was the highest during spring season with an average of 7.142 µg/l (Table 5). The maximum concentration of (∑PAH CARC was 23.352µg/l recorded at station 9 (El boghaz opening), (Table 2). Total hydrocarbon concentrations (THC= ∑ALIP+∑PAHs) were ranged from 407.60-1556.29µg/l with an annual average of 959.79 µg/l. This value is not safe and affect on marine organisms of the area. This is in accordance with conclusion derived from Mazmanidi et al. [26] who stated that the THC concentration in seawater, which can produce a harmful effect on the aquatic organisms, is about 50µg/l. Table 6 presents the monthly inflows of drainage discharges to Lake Burullus (106 m3) during 2006. The maximum content of drainage water was 423.7x106 m3 recorded during July 2006 (summer season), however the lowest content of 241.2x106 m3 was recorded during February 2006 (winter season). The annual average was ranged from 5.49 to 65x106m3 with a maximum of 20 and 18.5% recorded at Drains 9 and 11, respectively.

Table 1 Concentrations (µg/l) of hydrocarbons in water collected from Lake Burullus during winter 2006 Name 1 2 3 4 5 6 7 8 9 10 Naph 0.60 0.28 0.49 0.71 0.50 0.48 0.45 0.29 0.07 Acthy ND ND ND ND ND ND ND ND ND 0.04 Flu ND ND ND 0.57 ND ND ND ND ND 0.40 Chr ND ND ND ND ND ND ND ND ND 0.08 BaP ND 0.02 ND ND ND ND ND ND ND 0.05 DBA ND 0.03 ND ND ND ND ND ND ND ND B(ghi)P ND 0.08 ND ND ND ND ND ND ND 0.12 InP 0.08 ND ND ND ND ND ND ND ND ND ∑PAHs 0.683 0.414 0.494 1.274 0.501 0.480 0.451 0.285 0.073 0.681 ∑PAH CARC 0.084 0.050 NR NR NR NR NR NR NR 0.048 CARC% 12.31% 12.16% NR NR NR NR NR NR NR 7.02% ∑C10-C30 895.45 286.03 831.86 4484.28 568.93 3292.64 1555.73 842.00 772.35 1099.59 Pr 7.29 ND 2.95 7.34 1.56 42.88 13.09 1.10 480.23 ND Ph 10.88 3.16 14.08 46.93 10.23 38.20 60.68 21.86 166.30 ND ∑ALIP 913.61 289.18 848.88 4538.55 580.72 3373.72 1629.50 864.96 1418.89 1099.59 Pr/Ph 0.67 NR 0.21 0.16 0.15 1.12 0.22 0.05 2.89 NR Naph: naphthalene, Acthy: acenaphthylene, Ace: acenaphthene, Flu: fluoranthene, Chr: chrysene, BaP: benzo(a)pyrene, DBA: Dibenzo(a,h)anthracene, BghiP: benzo(ghi)perylene, InP: indeno(1,2,3-cd)pyrene, ND: non-detectable; NR: not recorded because one of the values equal zero; ∑ALP: aliphatic hydrocarbons; ∑PAH CARC : BaA+BbF+BaP+DBA+InP (IARC probable and possible human carcinogens); CARC: %PAHCARC/PAHs, Pr: pristine, Ph: phytane, ∑C10C30: total alkanes from C10 to C30.

Table 2 Concentrations (µg/l) of hydrocarbons in water collected from Lake Burullus during spring 2006 Name 1 2 3 4 5 6 7 8 9 Naph ND ND ND ND 0.793 ND ND ND 1.385 Acthy ND ND ND ND 0.538 ND ND ND 1.158 Ace ND ND ND 1.267 0.558 ND 0.279 2.032 0.100 Fl ND ND ND 0.027 0.259 11.332 4.607 ND 5.565 Phe ND ND ND ND ND ND ND 0.222 1.048 An 0.151 ND ND 0.008 ND ND 0.006 3.254 1.094 Flu 0.066 ND ND 0.330 0.131 0.038 0.030 0.457 2.832 Py ND ND 0.287 0.020 1.093 ND ND 1.043 0.751 BaA Chr BbF BkF BaP DBA B(ghi)P InP ∑PAHs ∑PAH CARC CARC% ∑C10-C30 Pr Ph ∑ALIP Pr/Ph

ND ND ND ND ND 0.337 0.962 1.926 3.442 2.263 65.74 160.44 5.594 30.006 196.042 0.186

0.062 ND ND 0.046 4.704 0.075 0.295 ND 5.182 4.841 93.42 245.85 34.159 67.989 347.995 0.502

0.134 ND 0.203 0.338 1.765 0.249 0.993 5.978 9.947 8.329 83.73 429.50 ND 68.349 497.846 NR

ND 0.364 0.050 0.209 0.183 0.025 2.016 0.621 5.120 0.878 17.15 254.19 0.518 ND 254.710 NR

0.315 0.491 0.396 0.120 0.151 0.049 0.088 0.110 0.897 0.215 1.984 1.235 0.084 2.504 0.177 3.930 7.463 20.025 3.524 5.921 47.22 29.57 214.7 84.35 ND ND ND ND 214.697 84.352 NR NR

ND ND ND ND 0.293 0.265 ND 0.096 5.576 0.654 11.73 98.75 0.770 ND 99.515 NR

2.544 0.265 0.180 0.049 0.505 0.394 0.344 7.449 18.738 11.072 59.09% 341.03 8.064 21.493 370.582 0.375

11.259 2.903 1.628 2.007 2.849 3.754 0.521 3.863 42.715 23.352 54.67% 157.17 ND 14.391 171.556 NR

10 0.840 ND ND ND ND ND 0.119 ND

11 0.531 ND ND 1.965 ND ND 2.487 0.214

12 ND 0.761 ND ND 0.078 ND ND ND

ND ND 0.091 ND 0.427 0.100 ND 0.129 0.139 0.016 0.738 0.095 ND 5.118 0.240 ND 6.715 1.436 ND 0.497 1.294 0.036 1.640 9.326 1.010 20.460 13.561 0.036 13.602 11.232 3.53% 66.48% 82.83% 106.23 574.22 127.98 ND 0.539 0.209 5.160 ND 0.667 111.391 574.761 128.854 NR NR 0.314

Table 3 Concentrations (µg/l) of hydrocarbons in water collected from Lake Burullus during summer 2006 Name 1 2 3 4 5 6 7 8 Naph 2.166 2.480 ND ND ND 0.578 1.964 2.412 Acthy ND ND ND ND ND ND ND ND Ace ND ND 0.045 0.016 ND 0.194 ND ND Fl 0.2254 ND ND ND ND ND ND 0.1890 Phe ND ND 0.129 ND 0.336 ND 0.300 ND An ND 0.317 ND 1.036 ND ND ND ND Flu ND ND ND ND ND ND ND ND Py 0.109 ND ND ND ND ND 0.165 ND ND ND ND ND BaA ND 0.361 ND ND ND ND Chr ND 0.010 0.145 ND 0.139 13.686 ND BbF 0.328 ND ND 0.010 0.084 2.616 1.662 ND ND BkF 0.158 ND ND ND 0.511 0.089 ND ND ND ND ND BaP ND 0.068 ND ND ND ND ND ND DBA 0.156 0.268 0.696 ND ND ND ND B(ghi)P ND 0.101 ND 0.355 ND ND ND ND InP ND 0.799 11.66 0.456 ∑PAHs ∑PAH CARC

∑C10-C30 Pr Ph ∑ALIP Pr/Ph

9 1.110 0.091 ND ND 0.378 3.634 25.494 4.956 10.382 0.686 0.432 1.944 0.25 0.226 0.574 ND

10 ND 0.117 ND ND ND 0.328 ND ND 0.051 ND ND ND ND ND ND ND

3.142

3.169

1.487

1.062

11.992

0.923

6.151

19.088

50.157

0.495

11 5.836 ND 4.510 ND 17.600 4.490 12.800 5.560 ND ND 3.160 1.930 2.780 2.520 6.220 0.602 68.008

0.485 2684.85 74.79 240.40 3000.04 0.31

0.361 53.92 5.66 ND 59.58 NR

1.067 13.15 ND ND 13.15 NR

0.01 96.63 ND 35.79 132.42 NR

11.656 228.93 ND 9.79 238.72 NR

0.152 302.49 45.83 ND 348.32 NR

3.072 325.42 138.85 164.34 628.62 0.84

2.358 174.17 ND 17.36 191.53 NR

11.29 238.42 11.63 280.33 530.37 0.04

0.0506 65.47 467.65 ND 533.12 NR

9.062 93.74 ND 59.22 152.95 NR

12 1.394 ND ND ND ND ND ND ND ND 1.120 0.216 ND ND ND ND

1.096 168.14 49.01 ND 217.15 NR

0.880 3.610

Table 4 Concentrations (µg/l) of hydrocarbons in water collected from Lake Burullus during autumn 2006 Name DL 1 2 3 4 5 6 7 8 9 Naph 0.04 ND ND ND ND 2.677 ND 1.110 ND ND Ace 0.01 ND ND ND ND ND 0.116 ND ND ND Fl 0.01 ND ND 0.303 0.032 ND ND 0.0002 ND 0.155 Phe 0.02 ND ND 0.188 ND ND ND ND 0.168 0.128 An 0.01 ND ND 0.042 ND 0.080 ND ND ND ND Flu 0.03 0.669 ND ND ND ND 0.132 ND ND ND Pyr 0.03 ND 0.018 0.184 ND ND ND 0.0002 ND ND BaA 0.04 0.019 0.010 ND ND 0.091 0.180 ND 0.088 ND Chr 0.04 ND ND 0.281 ND 0.328 0.057 0.0002 ND ND BbF 0.05 ND 0.026 0.038 ND 0.029 ND 0.0001 3.702 0.329 BkF 0.05 ND 0.009 0.531 ND 0.013 ND ND 1.467 0.152 BaP 0.05 0.052 0.178 1.494 8.849 ND ND 0.0004 0.038 ND DBA 0.06 0.066 0.217 0.023 ND ND ND ND 1.260 ND B(ghi)P 0.08 0.248 0.015 0.070 0.357 ND 7.543 ND ND ND InP 0.10 ND ND ND 0.055 ND 1.965 ND 0.418 0.130 ∑PAHs 1.055 0.471 3.154 9.294 3.217 9.993 1.111 7.142 0.893 ∑PAH CARC 0.138 0.430 1.554 8.904 0.120 2.145 0.0005 5.507 0.458 CARC% 13.06 91.22 49.29 95.81 3.71 21.46 0.04 77.11 51.29 ∑COMB NR NR 0.303 0.0321 2.6771 0.116 1.1099 NR 0.1545 ∑C10-C30 460.11 70.44 795.41 23.69 3674.1 287.27 2402.9 224.14 207.75 Pr 67.751 ND ND 5.514 ND 19.170 26.831 43.078 ND Ph 24.035 16.75 ND ND 11.381 ND 35.579 14.932 ND ∑ALIP 551.89 87.190 795.41 29.21 3685.4 306.44 2465.4 282.15 207.75 Pr/Ph 2.82 NR NR NR NR NR 0.75 2.88 NR DL = detection limit (µg/ml) for each individual PAH.

10 3.108 0.012 0.182 0.121 ND ND 0.044 ND ND ND 1.134 ND ND ND ND 4.601 NR NR 3.3017 387.84 14.601 ND 402.45 NR

11 0.206 0.645 0.146 ND ND 1.443 ND ND 0.069 0.111 0.507 0.063 ND ND ND 3.190 0.174 5.45 0.997 52.89 ND 10.82 63.71 NR

12 2.357 ND ND 0.041 ND ND 0.374 ND ND ND 0.069 ND ND ND 0.056 2.898 0.056 1.93 2.3571 7311.3 5.760 ND 7317.1 NR

Table 5. Seasonal distribution of individual PAHs and the % of different ratios for hydrocarbons in water samples collected from Lake Burullus during 2006 Compound winter spring summer autumn Average Naph 0.387 3.549 1.495 0.788 1.555 Acthy 0.004 2.457 0.017 0.064 0.636 Ace ND 4.236 0.397 0.068 1.175 Fl ND 23.756 0.035 0.054 5.961 Phe ND 1.348 1.562 0.010 0.730 An ND 4.513 0.817 0.187 1.379 Flu ND 6.489 3.191 0.052 2.433 Py 0.097 3.408 0.899 0.032 1.109 BaA ND 14.896 0.899 0.061 3.964 Chr 0.008 4.575 1.316 0.353 1.563 BbF ND 2.529 0.709 0.324 0.890 BkF ND 3.695 0.386 0.890 1.243 BaP 0.007 16.769 0.258 0.130 4.291 DBA 0.003 16.468 0.322 0.686 4.370 B(ghi)P 0.020 9.509 0.604 0.219 2.588 InP 0.008 35.041 1.199 3.918 10.042 ∑PAHs 0.534 153.240 14.107 7.836 43.929 Phe/An NR 0.299 1.912 0.054 0.529 Flu/Py NR 1.904 3.549 1.600 2.194 BaA/Chr NR 3.256 0.684 0.174 2.537 ∑PAH CARC 0.02 7.142 3.39 1.62 3.043 CARC% 3.75% 4.66% 24.03% 20.67% 13.28% ∑FPAH 0.391 33.998 1.944 0.975 9.327 ∑FPAH/∑PAHs 73% 22% 14% 12% 30% ∑COMB 0.143 119.242 12.163 6.862 34.602 ∑COMB/∑PAHs 27% 78% 86% 88% 70% ∑FPAHs/∑COMB 2.738 0.285 0.160 0.142 0.831 ∑ALIP 1555.76 254.358 503.831 1349.5 915.863 ∑THC 1556.294 407.598 517.938 1357.34 959.79 Thus the total annual average of drainage water is 32.5x106m3 (i.e. 32.5x109 l y ) from all drainage sources of Lake Burullus. The average annual concentrations of ∑THC were 959.8µg l-1. Therefore, the total annual inputs of hydrocarbon from all sources of drainage waters of Lake Burullus were 31 tons during 2006. Consequently, these annual inputs may affect the Mediterranean Sea by exchanging the water through El-Boughaz opening. -1

Table 6. Monthly inflows of drainage discharges to Lake Burullus (million m3) during 2006 Burullus Drain Gharbia Brimbal Month Tera East West 7 8 9 11 drain canal Jan 32.9 4.9 7.4 27.8 29.8 65 45.1 32.2 13.7 Feb 36.8 4.6 7.4 31.8 28.6 65 39.9 21.4 5.7 March 40.1 5.7 10.5 32.2 29.6 65 56.7 36.4 16.8 April 41.8 4.9 9 39.1 28.6 65 51.2 33.1 14.4 May 55.6 5 12.7 36.6 32.7 65 65.5 31.5 16.4 June 60.6 4.4 16.6 44.5 37.3 65 78.2 48.5 15.5 July 72.6 5.9 18.2 51.3 48.5 65 85 60.2 17 August 72.3 6.3 17.4 52 49.6 65 77.6 52.4 18 Sep 64.1 6.8 14.1 48.2 41.9 65 71.9 60 23.3 Oct 46.7 5.8 9.8 39.5 33.1 65 56.7 44 19.1 Nov 42.9 5.6 9.9 36.6 33.2 65 53.8 32.4 20.3 Dec 43.1 6 6.8 36.5 33 65 41.5 33.6 18.8 Annual 609.5 65.9 139.8 476 426 780 723 485.7 199 Average 50.792 5.4917 11.65 39.7 35.5 65.00 60.3 40.475 16.583 % 15.61 1.69 3.58 12.19 10.91 20.0 18.52 12.44 5.10 3.2 Sediment The ∑ALIP concentrations were varied between 31.34 to 331.45 ng/g (dry weight). Higher concentrations occurred at sites 2,3, 10 and 12. Lower concentrations occurred at sites 6, 8 and 9 (Table 7). The ∑n-alkanes concentrations (sum C10-C30) ranged from 31.34 to 331.45 ng/g (dry weight) was less than the recorded level for clean urban sites in Scotland, UK; with an average value of 3000 ng/g wet weight and ranged from 400-7100ng/g [27]. In addition, the recorded level for Black Sea ranged from 1200 to 240000 ng/g of sediment [28]. However, the present level of total nalkanes was higher than that recorded in sediment of the Suez Gulf ranged from 0.5 to 81.7 ng/g (dry weight), recorded by El Nemr et al. [29]. Total PAHs (∑PAHs) in sediments varied significantly among the studied locations. The values ranged from 388.8 to 19783.3 ng/g (dry weight). The maximum concentration of ∑PAHs was detected in samples of station 5 (in front of the outlet of drain 7). This drain discharges about 476 million m3 of untreated domestic wastewater with agricultural and industrial wastes into Lake Burullus. This is in accordance with El Nemr et al. [30] who stated that sediment samples collected near the sewage outlet, cities and harbor appeared to have extremely high concentrations of total PAHs. These suggest that PAHs accumulated in Mediterranean Sea sediments came from different sources such as sewage discharge from nearby human activities and fuel combustion emissions. The degree of sediment contamination by PAHs in this study is moderate in comparison with other aquatic systems in other countries (Table 8).

Table 7 Concentration (ng/g; dry wt) of hydrocarbons in sediment samples collected from Lake Burullus during 2006 Name 1 2 3 4 5 6 7 8 9 10 11 12 Naph ND 314.15 109.16 610.10 ND ND ND ND ND ND ND ND Acthy ND ND ND ND 85.71 2.07 7.29 11.00 19.79 ND 13.94 ND Ace 4.99 81.17 ND ND 65.27 47.50 ND 5.00 26.26 7.55 20.33 ND Fl ND 38.57 1.01 36.74 4.09 13.01 ND ND 2.82 ND ND 129.49 Phe 46.58 26.49 201.50 13.56 51.00 ND 212.46 42.20 23.22 ND 52.59 63.72 An 37.28 34.34 ND ND 29.31 ND 156.83 ND ND ND 53.79 176.57 Flu ND 38.07 ND ND 12.56 24.27 37.09 ND ND ND ND 142.83 Py 43.98 72.31 45.24 29.70 ND 68.76 ND 5.20 ND 18.75 2.37 60.00 BaA 164.00 41.09 ND 8.67 314.39 13.12 8.24 2.50 ND 10.96 3.73 ND Chr 57.78 49.35 96.34 224.42 784.84 167.13 39.95 ND 154.12 64.64 ND 43.84 BbF 21.27 176.03 74.73 83.78 136.75 ND 36.33 ND 32.06 14.54 ND ND BkF 687.66 1206.9 76.56 70.44 454.75 3293.1 94.27 10.00 55.68 179.64 ND 335.15 BaP 32.47 225.36 339.74 3.70 ND 249.94 46.83 1200.0 ND ND 390.50 ND DBA ND 216.80 ND 34.07 288.20 806.74 38.39 ND ND ND ND 837.98 BghiP ND 278.02 ND ND ND ND ND ND ND 279.86 ND ND InP ND 485.78 ND ND 17556 ND ND 3205.0 74.82 1822.0 428.22 ND ∑PAHs 1096.0 3284.5 944.3 1115.2 19783 4685.7 677.69 4480.9 388.78 2397.9 965.46 1789.6 80.69 331.44 139.12 129.64 36.21 31.34 89.72 39.55 45.78 119.91 66.54 218.48 ∑C10-C30 Pr ND ND 1.65 ND 75.78 ND ND ND 0.85 24.97 ND 0.68 Ph 56.42 ND ND ND ND ND ND ND ND ND 6.33 83.61 ∑ALIP 137.11 331.44 140.77 129.64 112.00 31.34 89.72 39.55 46.63 144.88 72.87 302.78 ∑THC 1233.1 3615.9 1085.0 1244.8 19895 4717.0 767.4 4520.4 435.4 2542.8 1038.3 2092.4

Table 8 Comparison of ∑PAHs and ∑AIP concentrations (ng /g; dry wt) in sediment measured in this study with those in other areas allover the world Area ALIP PAHs Lake Burullus, Mediterranean 31-331 389-19783 Egyptian Mediterranean coast 1.3-69.9 88-6338 Suez Gulf, Red Sea 0.52-88.38 158-10463 Kuwait, Gulf 200-280 Saudi Arabia, Gulf 100-1200 11000-6900000 Oman, Gulf 100-900 Crete, Eastern Mediterranean 500-5700 Western Mediterranean 180-3200 Safax Area, Tunisia 1121-5217 Gulf of Naples, Southern Italy 92-12561 Lazaret Bay, France 1600-48090 Gironde estuary, France 622-4888 Santander Bay, Spain 20-25800 Boston Harbor, USA 7300-358000 Washington coast, USA 29-460 Casco Bay, USA 16-21000 Kitimat Harbor, Canada ND-10000000 Sochi, Black Sea, Russia 700-3400 Hsin-ta Harbor, Taiwan 1155-3382 Kyenonggi Bay, Korea 9.1-1400 Beijing, china 14-4238 Xiamen Harbor, China 2900-61000

Reference this study [30] [29] [31] [32] [31] [33] [34] [35] [36] [23] [20] [37] [38] [39] [40] [41] [28] [42] [43] [44] [45]

3.2.3 Origin of PAHs in sediment Benzo(a)pyrene (BaP), the most potent carcinogenic PAHs, and the sum of six carcinogenic PAHs (∑PAH CARC ) [25] were highest at stations number; 5 and 8 with a concentration of 18296 and 4408 ng/g, respectively (Table 9). BaP was ranged from non-detected at stations 5, 9, 10 and 12 to 1200 ng/g at station 8 with a mean of 240.7 ng/g (Table 7), falling in the concentration range between rural and urban areas as mentioned by Menzie et al. [46]. The aromatic compound distributions differ according to the production sources, and on the chemical composition and temperature combustion of the organic matter [47]. One difficulty in identifying PAH origins is the possible coexistence of many contamination sources, and the transformation processes that PAHs can undergo before deposition in the analyzed sediments. Nevertheless, some compounds could exhibit comparable evolution kinetics that could be used to identify the origin of organic matter in the environment [13]. The compounds of Py, Phe and BbF are components of fossil fuels and a portion of them is associated with their combustion [48]. Benzo(a)pyrene is usually emitted from catalyst and non-catalyst automobiles. Benzo(a)anthracene and chrysene are often resulted from combustion of both diesel and natural gas [49]. The ratio of sum of major combustion specific compounds (∑COMB) to the sum of ∑16 EPAPAHs (∑COMB/∑PAHs) were ranged from 0.42 to 1.0 and the ∑COMB concentrations displayed values from 340 to 4623 ng/g (Table 9), representing average of 91% of total anthropogenic PAHs. This ratio was 1 at stations 1, 5, 6, 8 and 10, which indicated that the PAHs at these sites are mainly coming from combustion origin.

Table 9 Total hydrocarbons (∑THC), total aliphatic (∑ALIP), total PAHs (∑PAHs), pyrolytic PAHs ∑COMB), ( fossil PAHs (∑FPAH), carcinogenic PAHs (∑PAH CARC ), Phe/An ratio, flu/Py ratio for sediment samples collected from Lake Burullus during 2006 Site No. 1 2 3 4 5 6 7 8 9 10 11 12

Concentration, ng/g; dry weight

Ratio

∑THC ∑ALIP ∑PAHs ∑COMB ∑FPAH ∑PAH CARC Phe/An %COMB 1233.1 137.11 1096 1091.0 4.99 218.00 1.25 1.00 3615.9 331.45 3284.5 2850.6 433.89 1145 0.77 0.87 1085 140.77 944.3 834.1 110.17 414 NR 0.88 1244.8 129.64 1115.2 468.4 646.84 130 NR 0.42 19895 112 19783 19628 155.07 18296 1.74 0.99 4717 31.34 4685.7 4623.1 62.58 1070 NR 0.99 767.41 89.72 677.7 670.4 7.29 130 1.35 0.99 4520.4 39.55 4480.9 4464.9 16 4408 NR 1.00 435.4 46.627 388.8 339.9 48.87 107 NR 0.87 2542.8 144.88 2397.9 2390.4 7.55 1847 NR 1.00 1038.3 72.87 965.5 931.2 34.27 822 0.98 0.96 2092.4 302.78 1789.6 1660.1 129.49 838 0.36 0.93

NR: not recorded, because one of the values equal zero, %COMB: ∑COMB/∑PAHs. The sources of PAHs, where from fuel-combustion (pyrolytic) or from crude oil (petrogenic) contamination, may be identified by ratios of individual PAH compounds based on peculiarities in PAH composition and distribution pattern as a function of the emission source [13,24]. Ratio values such as phenanthrene/anthracene (Phe/An) and fluoranthrene/pyrene (Flu/Py) had been used by previous workers [19,20,21,50,51]. Petroleum often contains more phenanthrene relative to anthracene as phenanthrene that is more a thermodynamically stable tricyclic aromatic isomer than anthracene, so a Phe/An ratio is observed to be very high in PAH petrogenic pollution, but low ratio in pyrolytic contamination cases [24, 21,52]. Low Phe/An ratio values (10 were mainly contaminated by petrogenic inputs and Phe/An