Phuket Marine Biological Center Special Publication no. 19(1): 169-172 (1999) 169 EFFECTS OF EXPOSURE TO WATER-SOLUBLE FRACTIONS OF LUBRICANT OIL (MESRAN@ SAE 40 W) ON ATTACHMENT FORCE OF THE SNAIL LITTORARIA SCABRA (LITTORINIDAE : GASTROPODA) Markus T. Lasut & James Paulus Sub-Laboratory of Toxicology & Marine Pharmaceutics, Laboratory of Marine Sciences Faculty of Fisheries & Marine Sciences, Sam Ratulangi Uniuersity Jl. Kampus (Jnsrat Bahu, Manado 95115,lndonesia ABSTRACTAttachment force in gastropods has5 been used to measure responses of marine snailsLittoraria sgabra (shell length of 78-24 mm) to water-soluble fractions (WSF) of lubricantoil (Mesran@ SAE 40 W) in different concentrations. The ability to adhere to the substratumwas ineasured in two ways: 1) the weight required to lift up an animal when it attached toa substratum (termed tenacity). 2) the percentage of animals falling down from the wall ofthe experimental chamber (termed endurance). Higher concentrations of WSF and longerexposure times reduced the attachment force, both with respect to tenacity and endurance.This was related to the production of mucus as the first self-defence response ofZ. scabrato the oil. Production of mucus was the sublethal response, which required enerry thatfinally drained the resources available for attachment to the substrate. Statistical testsshowed that the effect of WSF on tenacity and endurance, and their interaction, weresignifrcant (p<0.05) in all experiments. INTRODUCTIONOiI is important for various refined B0-987o of the lubricants in use today (Bettonpetroleum product. InL992, the Institute of 1994). Saez et al. (1992) showed the negativePetroleum UK calculated that the crude oil effects of oil-derived hydrocarbons on theentering a refinery is converted primarily digestive gland of the snail Littorina littorea.to fuel (c.907o) while ca. 17o is converted to The ability to attach to a substrate islubricant oils (Betton 1994). Petroleum and essential for intertidal gastropods in relationits products enter the environment from to their activities (Imrie 1992) Attachmentseveral man-made sources: offshore could be used as a way to asses effects ofproduction, transportation, municipal and environmental pollution. We haveindustrial wastes. It is of global concern that investigated this possibility by studying thethe marine environment is vulnerable to oil attachment capability of the snail Zittorariapollution. scabra exposed to water-soluble fractions of Numerous reports are available, which lubricant oil. The ability to adhere to thedescribe the effects of oil pollution on marine substratum was measured in two ways: 1)environments and their organisms (Ahmad the tenacity measured as the weight 1988, Nybakken 1993). However, little is required to lift up an animal attached to aknown concerning of effects of lubricant oils substratum. 2) endurance measured as theincluding their water-soluble fractions percentage of animals falling down from the(WSF) on marine animals (Saezet al. L992). wall of the experimental chamber.The water-soluble fractions may comprise
170 Tlopical Marine Mollusc Programme QMMP) plastic chambers without lid (15 x 17 cm) MAIERIALS AND METHODS contained 0.8, 1.6, 2.4 PPm WSF and aL. scabrawas collected at Tongkaina, North control. Atotal of 60 snails (shell length 18-Sulawesi, Indonesia. The snails were stored 24 mm) were exposed to WSF fot t44 hoursin the laboratory fot a 2 days adaptation The number of animals falling down fromperiod before the experiment. All sea water the chamber wall was counted evety 24used was taken from the site where the hours. The measurements were done in 4specimens were collected. The salinity and replicates.temperature were 34 %o and 23-26 C The tenacity was analyzed by means ofrespectively. @ two-way ANOVA (Analyses of Variance) A lubricant for car engines (Mesran- while one-way ANOVA was aPPlied inSAE 40 W, produced bY PERTAMINA measurements of the endurance. A Ttrkey-Indonesia) was used as the test compound test was applied in both measurementsThe oil had been used in a car for up to 2000 (Sokal & Rohlf 1981; Fowler & Cohen 1990)km. The oil was mixed with sea water in achamber and left exposed to air for at least RESULTS30 days to release the water-soluble fractionThe WSF was separated from the water- Extraordinary amounts of mucus wereinsoluble fractions (WIF), which always form secreted by L. scabro exposed to WSF,the upper layer of the solution. The WSF indicating a stressed environment compared concentration was analYsed on a with the control. Further, the tenacity of spectrophotometer (wave length of 450 A) snails decreased as the concentrations and using t,t,t-Tbichloroethane as a standard the duration of exposure increased Gig 1) Three concentrations (08 1.6 and 24 ppm) of WSF were prepared by dilution with sea water. Chambers without lid containing 15 n oto* snails (shell length 18-23 mm) were exposed I 4ho*t for 0, 4 and 8 hours to the three I gtto* concentrations of WSF and a control Both concentration and duration ofthe exposure were variables of the test. After exposure to WSF, each animal was rinsed in clean sea water for 2to 3 minutes before being placed under a modifred wooden tool with glass as a substratum. Tenacity was measured in terms of the pull required to detach the animal from the substralum. A cotton line was attached to the snail. A small plastic chamber was fixed at the opposite end of the line running over conttol 0,8 1,5 24 a block. Water was added to the chamber Concennaion ofWSF ofoil (PPm) until the animal detached. The volume of water was measured (1 mI = 1 g) The Figure 1. Tenacity of individual L scobr6-exposed-to WbF prepared from lubricant oil (Mesran* SAE 40 W) measurements were done in 1-0 replicates The tenacity is expressed in terms of the weight (g) t Endurance was measured as the sd needed to pull the snails -of offthe substratum after 0, percentage of snails, which fell down frgm a 4, and 8 hrs e*posrrre to WSF at concentrations of ppm. The control was clean sea water chamber wall when they crawled up Four 0.8 to 2.4
Publication no 79(1): 169-172 (1999) 771 Phuhet Marine Biotogical center special had been used in a car engine forThe effect was statistically significant lubricant km prior to the test on the snails 2000(p<0.05). Interactiorr"r u"lr, conclerrttation up toand duration was also significantly (P<0.05). However the composition of the oil was notftt" "ffu"t of the WSF was significant analysed vv vvu F----(P<0.05)ataconcentrationof0.8ppmandTheWsFwasprobablytakenupbythe4hours of exposure.t/r1urwvr snails via the gills in connection with respitation (McMahon L992) Nelson-Smith Og72) iru Laws (1993) suggested that the looft--r-1-+-. toxic hydrocarbons apparently exert their ----. ;i;;;"p.rtbybecomingincorporatedinto ihtt makesup the interior of ^80 s -.-o ppm iftu futtv iuv"t As a result the1Tli,ll: cell membranes. ts l - -,-;;;, irarropt"aandceasestoproperlyregulate -_::::: F *l t^..- .- - substances between the ;;:;: tr" """i"ge of exterior of the cell I I tt -:r ini""iot utta zg I also interact with proteins in F *t ^ -.--....-.-. Hvdrocarbons plants and animals. Both a ,rariety of enzymes and structural proteins upp"?: t? 3 I r "- ^--.- jeneray ug"""d thut E ," I --*--^.-^ be affected. It is aromatic hydrocarbons are the most toxic | iotto*"abycycloalkanes olefins andlastly o l2o 144 ^Ikanes a (Laws 1993) zo 48 72 90 TIME (HouR) The first responses we observed in L scabra was a high production of mucus during exposure to ttre WSF Production of Figure 2. Endurance or L. scabra6::qT*::^HF *rr.oi drains energy of the snail (Davies ei orEou."a from lubricant oil (Mesranw SAE 40 w). Tle ot. tggZl. The decreasing ability to stick to il;;; tt expressed as the percentage of snails ifr"-r"frir"t"ttt (tft" teriacity test) or stay *hi.h *"r" ableio stay attached to the walls of tan *ul attached to the wall (the endurance test) containing from 08 to 24 ppm wsF The "o"ttol clean sea water. The was estimated every may be the visible signs of a drain of energy 24 hrs over a period of 144 hrs ""a"It"u""t ioi"rr"r, no relevant comparative studies unable to rhe endurance decreased all over time in :,il:"lr"Jili?:X1.:ltt}ir.u" concentrations,includingthecontrol(Fig-z) aLJ" .ffl*s of oil are not observed below Ho*"-n"" aft,er 144 hrs a total of 87Vo of the ;;;;;;;ions of 10 ppb. However, Gerlach tttuitt were still attached to the wall in the ;i;;il;;ested thai i pg/of water-soluble .o"t"of while onlv 67o marra$ed to do the ;;i;"-;";;;ts in water can harm sensitive 24 ppm same in the highest concentration of ;;;;;i*". as fish larvae hatching from fish on the endurance ; healthy. EPA (1976) inLaws (1993) from a ;; WSF The effect of the WSF of the snails was significant a reiuction in the chemotactic concentration of 1.6 ppm (P<0.05). ";i""""a of food by the snail Nassarius p""."ption DrscussroN * be ,n:lfutiT6tTltffii6i:lJi: gastropods was 1-100 mg/I The toxic effects of oil are believed to "ilydrocarbons.on Grazing mollusc are more tow-molec;i;r:;;igh Qp-CzD (Betton 1994) than filter-feeding mollusc caused by compounds, which are soluble in *uiJ" r aru Dvruv sensitive to oil which can separate oil from food particles (Laws 1gg3). The WSF in the present study may have (Cormack 1983) n""t tff""ted bV heating because the
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