Humic acids extracted from the sediment of Cochin inshore areas exhibited impairment of mercury

toxicity to marine algae maintained in Walne's medium under laboratory conditions. Cultures of

marine micro alga /50chrysis galbana contaminated with sublethal concentration of 10 ppb of inorganic

mercury and 10 ppm humic acids registered 18%, 21 % and 37% increase in oxygen production for

two, four and six hours incubation in light respectively, when compared to the controls. Similarly

the daily growth rate of seaweed Ulva lactuca registered 26% increase over the control when the

seaweed was maintained in medium containing 30 ppb Hg and 30 ppm humic acids

Kaladharan, P

Leelabhai, K S

CMFRI Digital Repository

Fishery TechnologJ) 2007, Vol. 44 (1) pp : 93 - 98 Effect of Humic Acids on Mercury Toxicity to Marine Algae P. Kaladharan* and K.S. Leela Bhai Central Marine Fisheries Research Institllte, P.B. No.1603, Cochin-682 018, India Humic acids extracted from the sediment of Cochin inshore areas exhibited impairment of mercury toxicity to marine algae maintained in Walne's medium under laboratory conditions. Cultures of marine micro alga /50chrysis galbana contaminated with sublethal concentration of 10 ppb of inorganic mercury and 10 ppm humic acids registered 18%, 21 % and 37% increase in oxygen production for two, four and six hours incubation in light respectively, when compared to the controls. Similarly the daily growth rate of seaweed Ulva lactuca registered 26% increase over the control when the seaweed was maintained in medium containing 30 ppb Hg and 30 ppm humic acids. The results are discussed in the light of employing humic acids as growth promoter of marine algae as well as abetting agent of mercury toxicity. Key words: Humic acids, mercury toxicity, abetting agent, seaweeds Humified organic matter, existing in coastal waters either as particulate or as dissolved form is collectively known as humic substances. The oceans and seas contain 2.6 x 1012 tonnes of' organic matter which approximately equal to the world's resources of coal or peat (Skopintsev, 1950) and the predominant form in which organic matter is encountered in natural water is as dissolved matter or water humus. They include humic acid, fulvic acid, humin and hematomelonic acid. They impart yellowish brown colour to coastal water, hence the name yellow substances. Humic acids are considered natural chelators (Saphiro, 1964) and the inhibitor of toxic effects of certain pollutants (Barbera el. a/., 1997; Ying ct. al., 1996). • Corresponding author: E-mail: kaladharan59®rediffmail.com Humic acids assume key role in coastal ecosystem as they induce phytoplankton production, nutrient transport and sediment formation (Perurnal, 1985). Spatial and temporal variation in humic acid levels have been reported from the mangrove forests of Pichavaram (Perurnal, 1985), inshore areas of Cochin (Kaladharan el. al., 1999a) and from the Arabian sea reporting the existence of a north-south gradient of organic carbon and humic acids (Sardessai el. a/., 1999). In the present study attempts have been made to estimate the spatial and temporal variation of total humic acids (THA, particulate + dissolved) from the estuarine as well as the inshore areas of Cochin, the levels of certain heavy metals adsorbed to THA of the sediment and the effect of THA on 94 mercury toxicity to micro alga lsoehrysis galbana and to a seaweed Ulvn Inc/uen . Materials and Methods Sediment samples were collected from inshore areas of Codlln on board R.V. Cndnlmin -IX from four stations, two in the esruarine and Table 1. Details of the sampling sites Station No. Position Depth(m) 1 9' 572 303 N,76' 172 003 E 6 2 9' 582 133 N,76' 142 503 E 8 3 9' 582 033 N, 76' 102 213 E 10 4 9' 582 443 N, 76" 062 433 E 20 another two in inshore regions. The station particulars are given in Table 1. Total humic acids were analyzed by the method described by Martin & Pierce (1971) and as modified by Hair & Basset (1973). Triplicates of sediment samples and the THA crystals extracted from sediment were digested at 120° c in a sand bath with cone. HNO, and H,O, (1: 4, v Iv) for 12 h until dissolution (Dalziel & Baker, 1983). All the digests were oxidized with potassium permanganate and potassium persulphate followed by hydroxylamine hydrochloride. Mercury was determined by cold vapour atomic absorption spectrophoto-metry (Hatch & Ott, 1968) using mercury analyzer (MA 5800 A, ECIL) with a detection limit of 0.02 ig. Prior to analysis, the instrument was tested for non-specific absorption. Remaining half of the THA digests were analysed for Cd, Pb, Cu and Zn using a AAS (Perkin Elmer, model 2380) in an air- acetylene flame with an accuracy within 10 % and the percent recovery of 94 %. Cultures of Isochrysis gnlbnnn and Ulvn Inctllen being maintained at the seaweed biotechnology lab of CMFRI (Kaladharan et nl., 1996) were utilized for the present srudy. LC 50 values (96 h, Probit analysis) of Ulvn Inctllen and lsoclm)sis gnlbmw for Hg was found to be 35 ppb and 25 ppb respectively . Sublethallevels(5, 10, 15 & 20 ppb for lsochrysis KALADHARAN AND LEELA and 5, 10, 20 & 30 PPb for lllvn) of inorganic mercury (HgCl,) were added to the culrures and incubated in 12 h light regime. The rate of photosynthetic oxygen released by the mercury treated lsoehrysis culrures with or without THA was srudied for a period of six hours and measured by the Winkler's procedure every two hour. Daily growth rate (DGR) of Ulvn lac/Ilen was calculated from the weekly gain/loss of wet weight from the initial weight of samples maintained in Walne's medium (Walne, 1974) using the formula where, Wl_ Weight (wet) gained or lost after culrure WO- Initial weight (wet) of samples prior to culture T - Dura tion of culrure in days Results and Discussion Total humic acids in the sediments of esruarine and inshore areas of Cochin showed maximum of 2.313 ppm during the postmonsoon in the esruarine (Stn I & II) and a maximum of 1.396 ppm during the SW monsoon months (Fig. 1) in the inshore area (Stn III & IV) . The observed levels agree well with the earlier report 3 1 [ 2 e ~ 15 • u E j' , 0.' o Premonsoon ",,"soon Postmonsoon Fig. 1 Humic acids in Cochin sediments EFFECT OF HUMIC ACIDS ON MERCURY TOXICITY TO MARINE ALGAE 95 Fig. 2 Metals extracted from the THA of sediments 80 70 60 E 50 "-Eo • '" 40 Q) > .330 20 • 10 0 Zn 200 E ~:~ 1 R 120 ..J f~~ l' 60 40 • 140 120 • • 20 o Zn (a) during pre monsoon • • • Inshore • Estuarine • • • Cd Pb Cu Hg (b) during S. W. monsoon • • •• , -.-. Cd Pb Cu Hg (c) during post monsoon • Inshore • • • Estuarine • • • • Cd Pb Cu Hg from Pichavaram mangrove swamps (Perumal, 1985) and from Inshore areas of Cochin (Kaladharan et al., 1999 a). Humic acids occur widely in various natural accumulations of organic matter including soils, peats, brown coals, sedimentary rocks and the range of recent subaqual deposits and the relative content of humic acids in sea sediment range up to 26 % (Bordovsky, 1964). Temporal variation of metals adsorbed to the THA of Cochin sediments are depicted in Fig.2. Levels of Pb and Zn were much higher than other metals studied. Cd and Cu were at lower levels both in the sediments from estuary as well as from the inshore regions and Hg remained the least. THA of marine sediment was known to contain high levels of Pb and Zn as in the sediment samples. However, levels of Hg were higher in THA extracted from the sediments than in the sediment itself both from the es tuary as well as from inshore areas (Fig. 3). Monsoon and Post monsoon seasons registered higher levels. of metals in the sediments than in sediment collected during the pre monsoon months. It is established that humic acids reduce the toxicity of tin (Sn II & Sn IV) towards a cyanobacterium, Synecocystis aquatilis incu1tures (Barbara et al., 1997). It is demonstrated that both the bioavailability and acute toxicity of Meothrin are decreased significantly by the presence of humic acids at the concentration of 5 and 10 ppm " '[ 08 j .3: 06 o I ") 02 D Estuarine 1 • Inshore Sediment THA of sediment Fig. 3 Levels ofHg in sediment and in THA extracted from the sediment 96 Table 2. Effect ofTHA on Mercury toxicity to lsochrysis galool/a Treatment Hg + THA (ppb)(ppm) 2hr % increase 0+0 252±0.076 5+0 2.5O±0.029 5+5 2.60± 0.018 104 10+0 2.30± 0.016 10+10 2.71±0.052 118 15+0 2.25±0.012 15+15 2.45±0.067 109 20+0 1.57±0.014 20+20 1.64+0.038 104 (Ying et al., 1996). The rate of oxygen production during photosynthesis by Isochrysis in the presence of sublethal levels of mercury at different time intervals influenced by the presence of THA are shown in Table 2. We observed that the mercury toxicity to IsochnJsis galbana could be impaired effectively (18 %, 21 % and 37 % for 2 hr, 4hr and 6 hrs respectively) by the THA up to 10 ppb of mercury; and beyond 10 ppb levels, the inhibitory effect of THA decreased considerably. Addition of THA beyond 20 ppm did not favour the oxygen production in IsochnJsis in culture (Table 2). However, the mercury toxicity to the seaweed Ulva lactuca (Table 3) was inhibited by THA up to 30 ppb (27 % higher than the control). Humic substances are known to form complex substances with metals( Barbera et al., 1997). This complex formation or binding with Hg may be one of the reasons for the inhibitory action of Table3. Effect of rnA on Mercury toxicity to Ulva ladi/ill Treatment Hg+THA (ppb)(ppm) 0+0 5+0 5+5 10+0 10+10 20+0 20+20 30+0 30+30 Growth rate (mg/ day) 69.17±0.97 62.98±0.968 60514±0.63 51.02 ±0.37 6O.04±1.20 43.19±1.81 54.47 ±0.56 30.33 ±0.65 38.23± 0.96 % increase 100 97 117 125 127 KALADHARAN AND LEELA 0, production (mI / l) 4hr % increase 6hr %increase 2.76±0.023 2.86±0.051 2.70±0.022 2.73±0.06 2.91±0.057 108 3.12±0.115 114 2.43±0.052 2.48±0.047 2.95±0.05 121 3.4±0.058 137 2.3O±O.045 2.3O±OO38 253±0.035 110 2.8O±0.047 122 228±0.032 2.31±0.039 2.48±0.031 109 2.64±0.032 114 THA on mercury. Greater effect of metal toxicity reduction (135 %) is reported (Rosko and Rachlin, 1975) due to chelation of citric acid and arninoacids on marine diatom Nitzchia c/osterium. Mercury pollution in India is going to be alarming if unchecked tha t India's annual mercury import between 1998- 2001 was 170-190 tonnes equivalent to 10% of total global consumption. Mercury consumption by the Indian chlor-alkali companies is at least 50 times higher than the average European consumption (Anon,2002). The total mercury pollution load in India is said to be about 125 tonnes of elemental mercury. Of this 40 % per annum is only due to operational loss of mercury to the environment by the existing mercury cell plants as a result of outdated technology they use. Certain hot spots of mercury contamination in the Arabian sea is reported by Sanzgiry et. al., (1988) and Kaladharan et. al., (1999 b). Mercury pollution would assume lethal proportions if efforts are not made soon to prevent its use. The increasing levels of mercury pollution in the coastal areas can be contained by certain bacteria (Magos et. al., 1964), by phytoplankton (Alavandi et. al., 1990) and also by THA as revealed by our present study. The help and assistance rendered by the crew, RV Cadn/mill IX , Shri A. Nandakumar and Shri L.R. Khambadkar in the laboratory and the encouragements received from Dr V.K.Pillai , Principal Scientist and Head FEMD (Retired) are gratehdly acknowledged. EFFECT OF HUMIC ACIDS ON MERCURY TOXICITY TO MARINE ALGAE 97 References Alavandi, S.V., Kaladharan, P. and Pillai, V.K. (1990) Vola tiliza tion of inorganic mercury by Isochrysis gnlbann Parke from aquatic system. Indinn J. Fish. , 37(2), pp163-165. Anon (2002) Down to Earth, 151h September 2002, pp27-28. Barbera, P .s., Daczorowska, R. and Skowronski, T. (1997) The impact of inorganic tin on the planktonic cyanobacterium Synechocystis nquatilis; The effect of pH and humic acid. Environ. Poll., 97(1-2), pp65-69. Bordovsky, O.K. (1964). Accumulation and transformation of organic substances in marine sediments. Mar.Ceol., 3, ppl-83. Dalziel, J. and Baker, C. (1983).Analytical methods for measuring metals by atomic absorption spectrophotometry, In: FAO Fish Tech. Pnp.,212, ppl4-20. Hair, M.E. and Bassett, C.R.(1973). Dissolved and particulate humic acids in an east coast estuary. Estunr. Coast. Mnr. Sci. , 1(1), pp107-111. Hatch, W. R. and Ott, W. L. (1968). Determination of sub micro gram quantities of mercury by atomic absorption spectrophotometry. Ani . Chem. , 40, pp2085- 2087. Kaladharan, P., Gopinathan, c.P. and Sridhar, N . (1996). Basic biochemical constituents in the laboratory cultures of six species of microalgae. In: The FOllrth Indiall Fisheries FOrll1ll Proceedillgs, 24-28 November 1996, Kochi, pp311-312. Kaladharan, P., Nandakumar, A. and Pillai, V.K. (1999 a). Total humic acids in the inshore waters of Cochin. In: Proc. Natn . Se m. Ocea. Fish nnd Fisheries, 1999, Irinjala- kuda, pp76-81. Kaladharan, P., Pillai, V.K., Nandakumar, A. and Krishnakumar, P.K. (1~99 b) Mercury in seawater along the wast coast of India. Indian J. mar. Sci. , 28, pp338-340. Magos, L., Tuffery, A.A, and Clarkson, T. W. (1964). Volatilization of mercury by bacteria. British J. Ind. med. , 21, pp294-298. Martin, D.F. and Pierce, R. A. (1971). A convenient method of analysis of humic acids in fresh water. Environ. Lett., 1(1), pp49-52. Perumal, P. (1985). Seasonal variations in humic acids in the Picha va ram Mangroves, South India. In: The Mangroves, Proc. Nat. Symp. Bioi. Util. Cons. Mangroves, Nov. 1985, pp369-371. Rosko, J.J. and Rachlin, J.W. (1975). The effect of copper, zinc, cobalt and manganese on the growth of the marine diatom Nitschia closterium. Bulletin of the Terrey Botanical Club, 102(3), pp100-106. Sanzgiry, S., Mesquita, A. and Kureishy, T.W. (1988). Total mercury in water, sediments and animals along the India coast. Mar. Poll. Bull. , 19, pp339-341. Saphiro, J. (1964) Effect of yellow organic acids on iron and other metals in water. J. Amer. Water Works Assn. , 56, pp1062-1082. Sardessai, S., Sarma, V.V.s.s. and Dileep Kumar, M. (1999) Particulate organic carbon and particulate humic material in the Arabian Sea. indian f. Mar. Sci., 28, pp5-9. 98 Skopintsev, B.A. (1950). Organic matter in natural water (water humus). Tr. CeDI. lnst. Acad. Nank S.S.S.R., 17(29). Walne, P.R. (1974).Cultures of bivalve mollusks, 50 Years' Experience at Conway. Fishing News (books) Ltd., England, ppl-173. KALADHARAN AND LEELA Ying, X., Wenzhong, W. and Yonyuan, Z. (1996). Effect of aquatic humic acids on Bioavailability and acute toxicity of Meothrin. Acta Hydrobiol. Sinica., 20 (2), pp160-163. Fishery TechnologJ) 2007, Vol. 44 (1) pp : 93 - 98 Effect of Humic Acids on Mercury Toxicity to Marine Algae P. Kaladharan* and K.S. Leela Bhai Central Marine Fisheries Research Institllte, P.B. No.1603, Cochin-682 018, India Humic acids extracted from the sediment of Cochin inshore areas exhibited impairment of mercury toxicity to marine algae maintained in Walne's medium under laboratory conditions. Cultures of marine micro alga /50chrysis galbana contaminated with sublethal concentration of 10 ppb of inorganic mercury and 10 ppm humic acids registered 18%, 21 % and 37% increase in oxygen production for two, four and six hours incubation in light respectively, when compared to the controls. Similarly the daily growth rate of seaweed Ulva lactuca registered 26% increase over the control when the seaweed was maintained in medium containing 30 ppb Hg and 30 ppm humic acids. The results are discussed in the light of employing humic acids as growth promoter of marine algae as well as abetting agent of mercury toxicity. Key words: Humic acids, mercury toxicity, abetting agent, seaweeds Humified organic matter, existing in coastal waters either as particulate or as dissolved form is collectively known as humic substances. The oceans and seas contain 2.6 x 1012 tonnes of' organic matter which approximately equal to the world's resources of coal or peat (Skopintsev, 1950) and the predominant form in which organic matter is encountered in natural water is as dissolved matter or water humus. They include humic acid, fulvic acid, humin and hematomelonic acid. They impart yellowish brown colour to coastal water, hence the name yellow substances. Humic acids are considered natural chelators (Saphiro, 1964) and the inhibitor of toxic effects of certain pollutants (Barbera el. a/., 1997; Ying ct. al., 1996). • Corresponding author: E-mail: kaladharan59®rediffmail.com Humic acids assume key role in coastal ecosystem as they induce phytoplankton production, nutrient transport and sediment formation (Perurnal, 1985). Spatial and temporal variation in humic acid levels have been reported from the mangrove forests of Pichavaram (Perurnal, 1985), inshore areas of Cochin (Kaladharan el. al., 1999a) and from the Arabian sea reporting the existence of a north-south gradient of organic carbon and humic acids (Sardessai el. a/., 1999). In the present study attempts have been made to estimate the spatial and temporal variation of total humic acids (THA, particulate + dissolved) from the estuarine as well as the inshore areas of Cochin, the levels of certain heavy metals adsorbed to THA of the sediment and the effect of THA on 

Effect of humic acids on mercury toxicity to marine algae

http://eprints.cmfri.org.in/7344/1/350-_FISHERY_TECHNOLOGY_2007.pdf

Effect of humic acids on mercury toxicity to marine algae

Abstract

Similar works

Full text

Available Versions

CMFRI Digital Repository