An Example from the Amoco Cadiz Oil Spill
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THE IMPORTANCE OF USING APPROPRIATE EXPERIMENTAL DESIGNS IN OIL SPILL IMPACT STUDIES: AN EXAMPLE FROM THE AMOCO CADIZ OIL SPILL IMPACT ZONE Downloaded from http://meridian.allenpress.com/iosc/article-pdf/1987/1/503/2349658/2169-3358-1987-1-503.pdf by guest on 27 September 2021 Edward S. Gilfillan, David S. Page, Barbara Griffin, Sherry A. Hanson, Judith C. Foster Bowdoin College Marine Research Laboratory and Bowdoin College Hydrocarbon Research Center Brunswick, Maine 04011 ABSTRACT: On March 16, 1978, the tanker Amoco Cadiz ran factors as possible in a sampling program to look at oil spill effects. aground off the coast of North Brittany. Her cargo of 221,000 tons of When using non-specific stress responses, it is important to be sure light crude oil was released into the sea. More than 126 miles of coast- just which Stressors are producing any observed responses. Multi- line were oiled, including a number of oyster (Crassostrea gigas j grow- variate statistical methods appear to be useful in associating body ing establishments. The North Brittany coastline already was stressed burdens of multiple Stressors with observed physiological effects. by earlier additions of oil and metals. In a field study by Gilfillan et al.,6 it was shown, using multivariate In December 1979, 21 months after the oil spill, measurements of techniques, that in populations of Mytilus edulis from a moderately glucose-6-phosphate dehydrogenase activity, aspartate aminotransfer- polluted estuary the animals were responding to Cu, Cr, and Cd at the ase activity, and condition index were made in 14 populations of C. same time. gigas. Five of the populations were outside the oil spill impact zone; In the present study, specific activities of glucose-6-phosphate de- nine populations were within it. At the same time the body burden of hydrogenase (E.C. #1.1.1.49) and aspartate aminotransferase (E.C. Cr, Ag, Pd, Zn, Cu, Cd, and Hg, as well as the body burden of #2.6.1.1), as well as condition index (dry weight of tissue/total aliphatic and aromatic hydrocarbons, was determined. Also at the weight x 100), were determined as indices of physiological well being. same time, the numbers of total and fecal coliform bacteria in the Glucose-6-phosphate (G-6-P) is important in the metabolism of car- ambient water were determined. bohydrate; aspartate aminotransferase (AAT) is important in the Results of stepwise multiple regression have shown that 21 months metabolism of protein. Condition index (CI) is taken as an integrated after the Amoco Cadiz oil spill, aromatic hydrocarbons were only one measure of the animals' nutritional status. of three factors adversely affecting C. gigas populations in North Brit- Because the Amoco Cadiz oil spill impact zone was clearly not tany. Multiple regression techniques can be extremely useful in identi- pristine before the spill, a number of potential Stressors were mea- fying those Stressors associated with physiological effects in populations sured. These were chosen on the basis of personal observation and of animals. information derived from the literature. Potential Stressors measured included body burden of Cr, Ag, Pb, Zn, Cu, Cd, and Hg, as well as body burden of aromatic and aliphatic hydrocarbons. Also deter- mined were the numbers of total and fecal coliform bacteria in the ambient water. The tanker Amoco Cadiz ran aground off the coast of Brittany, Physiological indices and Stressor levels were determined for 14 France, on March 16,1978. Her cargo, 221,000 tons of light crude oil, populations of C. gigas from the North Brittany coast. was released to the environment over the next 15 days.8 More than 126 miles of coastline were impacted. Within the impacted zone were a number of oyster culture establishments. The dominant species of oyster grown in these establishments was Crassostrea gigas. Materials and methods The North Brittany coastal environment was known to be subject to pollution from a number of sources, including metals, hydro- Sampling location and methods. The locations of the 14 populations carbons, and sewage.1 As a result the oysters could be expected to be of C gigas are shown in Figure 1. Nine populations were from within responding not only to hydrocarbons from the Amoco Cadiz, but also the Amoco Cadiz oil spill impact zone; five were not affected by the to pollutants from other sources. Amoco Cadiz oil spill. All populations were sampled during the sec- The measurement of various physiological parameters in marine ond and third weeks of December 1979. Oysters were collected from animals as biochemical sublethal stress indicators has been discussed the natural environment at each location. They were wrapped in clean in a recent review.9 Ideally, a biochemical stress indicator should aluminum foil and held frozen (-18° C) pending analysis. relate to a given Stressor in a dose-response relationship, as has been Coliform bacterial analysis. Samples of ambient water were col- 3 demonstrated in a number of systems. This approach breaks down in lected in one-liter, clean glass bottles and held ice-cold. These water the field situation, where there is no overwhelmingly large input of a samples were analyzed for total and fecal coliform bacteria content by single Stressor or pollutant. the membrane filter technique.2 Results are reported as the number Animals and plants respond to their entire environment and not of viable bacteria per 100 ml of ambient water. just to any petroleum that may be present. Oil spills seldom occur in Tissue metals analysis. Concentrations of Cr, Ag, Pb, Zn, Cu and pristine environments. Therefore, it is essential to include as many Cd were determined in the tissue samples using flame atomic absorp- 503 504 1987 OIL SPILL CONFERENCE I " JERSEY é¡j>¿¿? ψ:;: à ENGLISH CHANNEL fe !2 IO ,Tregastel *L· liedeBrehat ||2,3 P MANCHE , / \ I / eWiy^/r-^-J^Pφimpol / V.: AMOCO / Rosc0ff^V / ^•';-' ·« i %;; ^LΏ^^^ . -k -^^tMa,<*^ Ouessant /portsali '' 9 J '' vKly:'." ' ' % V ;"\J Downloaded from http://meridian.allenpress.com/iosc/article-pdf/1987/1/503/2349658/2169-3358-1987-1-503.pdf by guest on 27 September 2021 MiedeMolene^jh-^ FINISTERE COTES-DU-NORD { ) VT^\. ^^'V·.. -N y· i ILLE-ET-VILAINE I v · V-feA-r^iconcarneau / ·. / ¿¿•'^^^\x::r.:yi.m.- :¿ ..· MORBIHAN \Λ / ^v7 ^^^4 ^^^;:;AΏ^ ^ /' Ώ, FS·» <^Hv · /^: XP,nnes \-*" *"" ' — Boundry of Brittany \^0^%J^^ LOIRE-ATLANTIQUE Department Boundry ^ ^ ¿Ö^· ■ . \ ."·" .—' 0 10 20 30 \^g <Á£Á·^^^ /' Km. ^^^^^I^;·/^ \ ·■-·.·.; ^'* J^& " [ ' <^!v"·« 1 Figure 1. Map of North Brittany showing sampling locations—1-3, Cancale; 4, lie Tudy; 5, Trieux River; 6, Dourduff; 7, Keramel; 8, Pen ar Lan; 9, Ty Nod; 10, Carantec; 11, St. Pabu; 12, Paluden; 13, Baie des Anges; 14, Anse de Broennou. Sampling locations 1 to 4 are not within the oil spill impact zone. The Trieux River sample was from an oyster park that was not oiled by the Amoco Cadiz oil spill. tion spectrophotometry on nitric acid digestates of the wet tissue The hexane eluate contained the aliphatic (nonpolar) fraction of hy- samples. Mercury was determined spectrophotometrically on a sub- drocarbons in the sample. Elution with 50/50 hexane/methylene chlo- sample of the digestate using the cold vapor technique. ride yielded an "aromatic" (polar) fraction of hydrocarbons contain- Each field sample consisted of at least five oysters that were sub- ing any aromatic hydrocarbons present plus other substances that sampled on a random basis in triplicates for chemical analysis and behave as aromatics in the liquid chromatography procedure. Each physiological measurements. Frozen subsamples of each of the tripli- fraction was quantified by weighing in tared containers on a micro- cate oyster homogenates used for the enzyme analyses were thawed balance. and a weighed aliquot was taken for dry weight determination. The The aliphatic fraction was analyzed by injection of a known aliquot samples were then weighed and digested in three aliquots (10 ml/10 of the sample into a Perkin-Elmer 3920B flame ionization detection ml/5 ml) of concentrated nitric acid, with the digestate taken to dry- (FID) gas Chromatograph equipped with an MS-41 sample injector, ness after the addition of each aliquot. The residue was dissolved in using a 15 m by 0.32 mm ID fused silica capillary column coated with 25 ml of 5 percent v/v nitric acid and stored in acid-washed containers SE-30. The instrument was further modified at the detector to allow pending analysis. The results of the analyses are expressed as parts the end of a capillary column to be inserted directly into the FID jet. per million on a dry weight of sample basis. The aromatic fraction was similarly analyzed on a Perkin-Elmer The recovery of the method was determined using homogenates Sigma 3B GC (FID), also equipped with an MS-41 injection acces- spiked with known amounts of the metals. With the exception of Ag, sory, using a 15 m by 0.32 mm ID fused silica capillary column coated all of the metals tested gave recoveries in the range of 70 to 95 percent. with SE-52. Tissue hydrocarbon analysis. Oyster tissue samples were extracted The gas chromatography conditions were the same for both instru- by refluxing a homogenized sample of 60 to 100 grams of shucked ments. The carrier gas was helium with a linear velocity of approxi- meats with 2 M KOH in methanol for three hours to extract and mately 40 cm/s at 80° C. The temperature program was 80° C for saponify the tissues. The cooled homogenate was centrifuged to re- 1 minute, then 4° C/minute to 280° C, with a final hold at 280° C for move solid matter and the supernatant was extracted with three por- 16 minutes.