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Two Freshwater Fish Species (Liza JOURNAL OF INTERNATIONAL ACADEMIC RESEARCH FOR MULTIDISCIPLINARY Impact Factor 1.625, ISSN: 2320-5083, Volume 2, Issue 12, January 2015 TEMPORAL AND SPATIAL VARIATIONS OF TOTAL PETROLEUM HYDROCARBON CONCENTRATIONS IN TWO COMMERCIAL FISH SPECIES FROM AL-KAHLAA RIVER IN MISSAN GOVERNORATE/IRAQ PROF. DR.HAMID.T. AL – SAAD* SALIH.H. JAZZA** ABDUL -HUSSAIN.Y. AL -ADHUB*** *Marine Science Center, University of Basrah, Iraq **University of Missan /College of Science, Iraq ***University of Basrah /College of Science, Iraq ABSTRACT ` Two freshwater fish species (Liza abu and Carassiusauratus) were collected monthly, during the period November 2012 to October 2013 from four different sites distributed along Al–kahlaariver in Missan province( AL-Magideh, Treatment unit, AL-Husaichi and AL- Zubair), in addition to a reference station which lie on the Tigris river close to the Amarh district (about 25Km). Concentrations of Total Petroleum Hydrocarbons in the muscles of these species were determined spectrofluorometrically. The highest concentrations of L.abu species 37.3 µg/g.dry weight was recorded during the summer in the treatment unit station, while the lowest concentrations 4.81µg/g. dry weight was recorded during the spring in Reference station. Whereas for C.auratus species the highest concentrations 29.303µg/g.dry weight was recorded during autumn in the treatment unit station, while the lowest concentrations 2.51µg/g.dry weight was recorded during the spring in Reference station. Also, we found that L.abuspecies have more ability to accumulate TPHs in their tissues compared with C.auratus species. The levels of TPHs in these fishes were high considering the recommended value from the World Health Organization which is from 0-0.001 µg/g for human consumption. This indicated that the study area was generally contaminated with hydrocarbons and continuous consumption of this food may pose public health hazards. Introduction: Total petroleum hydrocarbon(TPH) is a term used to describe any mixture of hydrocarbons that are found in crude oil. They are toxic compound are relatively soluble in water. As a result they are serious aquatic hydrocarbon pollutants (Herasetal,1992).AL-Saad and AL-Asadi(1989)reported that some fishes have different abilities to accumulate certain pollutants from the aquatic environment. Fish may be exposed to spilled oil in different ways. They may come into direct contact and contaminate their gills, the water column may contain toxic and volatile components of oil that may be absorbed by their eggs, larvae, and juvenile stages and they may eat contaminated food and accumulate toxins in their adipose tissues 439 www.jiarm.com JOURNAL OF INTERNATIONAL ACADEMIC RESEARCH FOR MULTIDISCIPLINARY Impact Factor 1.625, ISSN: 2320-5083, Volume 2, Issue 12, January 2015 especially aromatic compounds with higher concentrations in the brain, glands and gonads.These compounds accumulate with lower concentrations in the gills,liver and muscles because of the presence of high amounts of fats in these organs. Fishes that are exposed to oil may suffer from changes in heart and respiratory rate, enlarged livers, reduced growth, fin erosion, a variety of biochemical and cellular changes, reproductive and behavioral responses. Chronic exposure to some chemicals found in oil may also cause genetic abnormalities or cancer in sensitive species (Ackmanet al.,1996 ; AL-Saadet al., 1997 ; Deb et al., 2000).The hydrocarbon pollutants have indirect effects on fishes as they cause the destruction or death of aquatic grasses,aquatic plants,coral reefs,zooplankton and phytoplankton which are used as a source of protection for the growth of larvae and small fishes or as a food for this animals(GESAMP,1993 ; AL-Saadet al., 2003). Reynaud and Deschaux (2006) stated that fishes are quite sensitive to hydrocarbon compounds, leading to several specific and non- specific response by their immune system. The specific responses may involve the production of antibodies, and the unspecific responses may involve effects on increased activities such as lysozyme and/ or phagocytosis. Present study provides information on temporal and spatial variations of TPHs in two commercial fish species from AL-Kahlaa River in Missanprovince. Materials and Methods: Collection of Samples Fish samples were collected monthly during the period November 2012 to October 2013 from Four stations namely AL-Magideh, Treatment unit, AL-Husaichi and AL-Zubair distributed along Al –kahlaa river with reference station which lies on the Tigris river close to the Amara district about 25 Kmas shown in Figure 1. Samples were collected by using the trawl net, washed thoroughly in the river water and were placed in aluminum foils before being transferred to the lab ice –packed. The samples were kept under refrigeration prior to analysis. Classification of fish species. Family: Mugilidae Sp:Liza abu Family: Cyprinidae Sp: Carassiusauratus 440 www.jiarm.com JOURNAL OF INTERNATIONAL ACADEMIC RESEARCH FOR MULTIDISCIPLINARY Impact Factor 1.625, ISSN: 2320-5083, Volume 2, Issue 12, January 2015 Preparation of fish samples Fish samples were rinsed thoroughly with distilled water in the lab, their muscles were cut in to small parts and dried in the freeze – dryer. These were then grounded and sieved using a 63 μ metal sieve and then placed in a clean glass vial, ready for analysis. Extraction of hydrocarbon compounds from fish muscles. The procedure of Grimalt and Oliver (1993) was used for the extraction of hydrocarbon compounds from fishes. Five grams of dried fish muscles were placed in a pre–extracted cellulose thimble and soxhlet extracted with 150 ml methanol: benzene ( 1 : 1 ratio ) for 24 - hours. They were then saponified for 2 - hours with a solution of (4 N) KOH. After that, the extract was left to cool and placed in a separating funnel, 50 ml of n-hexane was added to the separating funnel, the content were mixed and the mixture was left for some time until it was stable forming two layers. The unsaponified layer which contains hydrocarbons was obtain and passed through a column chromatography filled with glass wool at the bottom followed by a layer of silica gel ( 100 – 200 mesh ), a layer of alumina (100 – 200 mesh ) and then of anhydrous Na2So4 was placed to isolate the aliphatic fraction, 50 ml of benzene was added to isolate the aromatic fraction.These fractions were reduced to a suitable volume prior to analysis. Spectrofluorometershimadzu RF – 540 equipped with recorder type Shimadzu DR- 3 was used to determine total petroleum hydrocarbons in fish samples. The basis quantitative measurements were made by measuring thee mission intensity at 360 nm with excitation set at 310 nm. TPHs were measured after calibrating this apparatus.Standard solutions prepared from crude oil was obtained from Missan Oil Company. Figure ( 1) : map of AL-Kahlaa river showing the position of stations. 441 www.jiarm.com JOURNAL OF INTERNATIONAL ACADEMIC RESEARCH FOR MULTIDISCIPLINARY Impact Factor 1.625, ISSN: 2320-5083, Volume 2, Issue 12, January 2015 Results and discussion: Using fish muscle is very important for the determination of levels of hydrocarbons and due to the analysis of their levels in muscles would give more information about petroleum hydrocarbon (Melancon and Lech, 1987; Hantoushet al.,2001).Results of this study summarized in figures (2 and 3) revealed that the mean concentrations of TPHs (µg/g) in muscles of C. auratus species during four seasons ranged from 3.67 to 8.88 during the winter, from 2.51 to 2.51 during the spring, from 9.03 to 24.46 during the summer and ranged from 8.63 to 29.303 during the autumn,while in muscles of L. abu the mean concentration ranged between (6.3-10.52 ), (4.81-21.12), (16.2-37.3) and (9.91 -32.71) during the winter,spring,summer and autumn respectively.Therefore ,results of statistical analysis in tables (1,2) showed that there was a significant seasonal and spatial variations. These levels of TPHs indicate that the fishes found at AL-Kahlaa river have higher TPHs values.This means that these animals have accumulated more hydrocarbons indicating a higher levels of pollution in these areas which comes from surrounding cities into the river,therefore transported into the tissues of these fishes via water intake (Jack et al.,2005).Ability of these fresh water fishes to accumulate TPHs varies and follow the order L. abu>C. auratus this is due to different lipid content, food habit, temperature, size, weight, age and sex of these fishes (Hellouet al 1993; Quintero and Diaz,1994 ; Nasir,2007 ;AL-Shwafi, 2008).Since L. abu and C. auratus constitute the major food fish of the population of Missan, the danger of bio-accumulation of TPHs in humans through the food chain is a great possibility and need to be checked. Therefore elevation of their levels in these commercial fishes calls for concern because the level of TPHs in these fishes are high considering the recommended value from WHO which is from 0-0.001 µg/g for human consumption. This indicated that the study area was generally contaminated with hydrocarbons and continuous consumption of food from this area may pose public health hazards (FMENV,1992). There is therefore the need to develop a management plan to ensure that petroleum hydrocarbon contamination of this area is prevented in order to achieve good aquatic life and to avoid possible health problems through the consumption of contaminated fresh water resources such as fishes by the local communities. From table (3) we noticed that the levels of TPHs in two fish species in AL- Kahlaa river were within the extent which were recorded in other areas from Iraq such as Shatt AL-Arab, Khor AL-Zubair, North West Arabian Gulf and Iraqi marine waters but their levels were more than those recorded in fishes from Iraqi southern marshes (Hor AL – Howaiza and Hor AL–Hammar) and the Euphrates river.
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