-t/Phuket Marine Biological Center Special Publication no. 19(1): 165-168 (1999) 165

EFFECTS OF SALINITY-CYANIDE INTERACTION ON THEMORTALITY OF ABALONE HALIOTIS UARIA

(IIALIOTIDAE : GASTROPODA)

Markus T. Lasut.Sub-Laboratory of Toxicology & Marine Pharrnaceutics, Laboratory of Marine Sciences.Faculty of Fisheries & Marine Sciences, Sarn Ratulangi Uniuersity, JI. Kampus Unsrat

Bahu, Manado 95 11 5, Indonesia

ABSTRACTEffects of salinity (25-35 %o) interaction with cyanide (KCN and NaCN) on survival havebeen tested on abalone Haliotis uaria. Highest mortality occurred at a salinity of 25 %o

and 4 ppm of both forms of cyanide. The lowest salinity could have caused stress, makingabalone more sensitive. The effect of cyanide on survival was significant at 1 ppm of KCNand 2 ppm of NaCN Q?<0.05), while the salinity and the interaction with cyanide were not(p>0.05). Seemingly, the KCN was moretoxic than the NaCN.

INTRODUCTIONCyanide (CN-) occurs in marine environ-ments in combination with ions of salinewater, forming potassium sait cyanide acid(KCN) and sodium salt cyanide acid (NaCN)compounds. Cyanide also exists as HCN, avery toxic compound. It is naturally foundin the marine waters in very lowconcentrations. However, its concentrationmay increase due to pollution by cyanide-containing waste.

Cyanide is widely used in industries toextract metals, such as gold from ores, inmetal refining, metal cleaning andelectroplating operations, and in certainmineral processing operations.Around the Minahasa Peninsular (NorthSulawesi, Indonesia) the concentration ofcyanide has probably increased due to acontribution from a gold mining company.No specific measurements are available butthe company has dumped a hundred ton perday of cyanide containing wastewater to thesea below the tropical thermocline(approximately 82 m depth) through apipeline (Anonymous 1994). However, thedepth of the tropical thermocline is a matterof opinion because no precise data areavailable.

The objective of the present study is toinvestigate the effects of salinity-cyanide(KCN & NaCN) interaction on the mortalityof the intertidal abalone Haliotis uaria.Abalones have rarely been used as testorganisms for toxicity tests, although theyhave been used to study the interactiveeffects of pentachlorophenol and hypoxia(Tjeerdema et al. t99la), the sublethaleffects of pentachlorophenol (Tjeerdema efal.799lb), the effects ofgas supersaturationon the behaviour, growth and mortality(Leitman 1992), the interactive effects ofpentachlorophenol and temperature(TJeerdema et al. 1993), and the effects ofsalinity and diazinon (Kaligis & LasutL997).

MATERIALS AND METHODSH. uaria were collected from January toApril 1997 along the Likupang Beach,Minahasa, North Sulawesi. The abaloneswere kept in the Marine SciencesLaboratory. In accordance with Kaligis &Lasut (1997) they were acclimated for 2 to5 days at room temperature (24-25'C). Theseawater (salinity 33-34 %o), was well-aerated and unfiltered. It was changed

166 Tbopical Marine Mollusc Programme (TMMP)

every 24 hours. Abalones were fedmacroalgae Gracilarlo sp. during thisperiod. The pH was about 8 when measuredbefore and after the experiment. No foodwas supplied during experiments.Abalones with shell lengths ranging from33.12-50.18 mm were prepared for eachexperiment conducted in 3-litres plasticbowls, which contained 1 litre of seawater.Three abalones were put in each bowl. Thetoxicity of KCN and NaCN in four differentconcentrations (1, 2 and 4 ppm plus onecontrol without cyanide) was tested inrelation to 3 salinities (25, 30 and 34 %").The cyanide concentrations were based ona preliminary test, which showed that7.76and 9.45 ppm were the median lethalconcentration (LCSO) for 96 hours of bothKCN and NaCN. Abalones were scored asbeing dead according to the criteria bySinghagraiwan et al. (7992) and Kaiigis &Lasut '1997).A two-way ANOVA (Sokal & Rohlf 1981;Fowler & Cohen 1990) was applied toanalyse if different concentrations ofsalinit5r, cyanide and their interaction wereaffecting the mortality.

RESULTSNo mortality occurred in any of the controls.

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Concentration of KCN {ppm)

Figure 1. Interactive effect between salinity andcyanide (KCN) on the mortality of the abaloneH. uaria along 96 hours of experiment.

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0't 24Concentration of NaCN (ppm)

Figure 2. Interactive effect between salinity andcyanide (NaCN) on the mortality of the abaloneH. uaria along 96 hours of experiment.

At a salinity of 25 Voo, 1007o mortality wasrecorded in L,2, and 4 ppm (Figs. 1-2). Theresults of the statistical test (ANOVA)showed that the effects of KCN to themortality was significant at a concentrationof 1 ppm (p<0.05), while the salinity andthe interaction between salinity and KCNwere not affecting mortality (p>0.05).In the interactions between 1,2 and 4 ppmof NaCN and 25 %o S, tlne mortality was 11,44 and 66 7o respectively. In l, 2, and 4 ppmof NaCN and 34 %o S, mortality was 11,22and 44 % respectively. The NaCN from aconcentration of 2 ppm was significantlyaffecting the mortality (p<0.05), while thesaiinity and the interaction between salinityand NaCN were not affecting mortality(p>0.05).

DISCUSSIONKaligis & Lasut (1997) reported that thecombination between salinity and theinsecticide diazinon resulted in mortality ofthe abalone H. uaria; especially in lowsalinity (25 %"). The same trend was foundin the present study. Mortality was highestin25 %o and the mortality depended on theconcentration of KCN and NaCN (Figs. 1&D.

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phuhet Marine Biological Center Special Publication no. 19(1): 165-168 (1999) 167

H. uaria is thriving in salinities of 32'5 to35 %o (Fuse 1981). The abalone is mode-

rately tolerant to seawater of 25 %o S

without acclimation and to a salinity of 20

7oo S wit]n acclimation (Kaligis & Lasut7gg7). However, 25 %" S is lower than theoptimum and may cause stress making thespecies more sensitive to pollutants.KCN was relatively more toxic than NaCN'The biochemical action of cyanide is that itdeprives the body ofoxygen by acting as a

chemical asphyxiant. Cyanide inhibits an

enzyme involved in the oxidative phospho-rylation by which the body utilises oxygen'It has been found that mitochondriadeprived ofoxygen fail to show significantoxidative phosphorylation (Edwards &Hassall 1980; Manahan 7992)' Theinhibited enzyme is ferricytochromeoxidase, an iron-containing metalloproteinthat acts as a frnal acceptor ofelectrons.Cyanide bonds to the iron (III) of theferricytochrome enzyme, preventing itsreduction to iron (II). The result is thatferrouscytochrome oxidase' which isrequired to react with oxygen, is not formedand utilisation of oxygen in cells isprevented., Ieading to rapid cessation ofmetabolic processes (Edwards & Hassall1980; Manahan 1983; Bohinski 1987;Manahan t992).

Brachet (1957) reported that cyanideinhibited respiration in developing sea

urchin eggs. He, furthermore, stated thatmany cytologicai abnormalities were found,

such as signs of degeneration of thechromosomes. Lasut & Lintong (1998;

unpublished data) found that larvae of sea

urchins were malformed if the eggs hadbeen exposed KCN.Efforts to reduce the toxicity of cyanide inan effluent can be done bY alkalinechlorination or by catalytic oxidation'However, cyanide wastewater containingnickel or silver is difficult to treat byalkaline chlorination because of the slow

reaction rate of these metal complexes(Lankford 1990).

ACKNOWLEDGEMENTSSpecial gratitude is extended to Prof. Dr. R'M. Rompas for valuable advise. I want to

thank mv colleague Mr. M. R' Badu for hisassistance in all phases of the work' I amvery indebted to the Tropical MarineMollusc Programme (TMMP) sponsored byDANIDA for the chance to present thispaper in The Ninth International Workshop/Congress of Tropical Marine MolluscProgramme (TMMP),August 19-29, 1998 atLombok, Indonesia.

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