J O U R N A L

O F

C O A S T A L

S C I E N C E S

JOURNAL OF COASTAL SCIENCES

Journal homepage: www.jcsonline.co.nr ISSN: 2348 6740 Volume 1 Issue No. 1 - 2014 Pages 27-32

An assessment of bacterial and biochemical characteristics in the placer mineral rich zones of southern coastal region, Tamil Nadu, India
V.K. Hridya*, K. Priyadharsini, Prince S. Godson, N. Chandrasekar
Centre for Geotechnology, Manonmaniam Sundaranar University, Tirunelveli, Tamil Nadu 627 012, India

ABSTRACT

ARTICLE INFO
Received 25 November 2013 Accepted 2 March 2014 Available online 6 March 2014 Keywords BiochemicalComposition, Marine sediments, Microbial, Placer minerals

`
The samples collected from Kanyakumari to Inayam on the coastal belt of low, mid and high tide regions of Tamil Nadu were studied for the distribution of biochemical variables such as carbohydrates, proteins, amino acids and lipids. This study area is a potential mining site of Placer Minerals. Carbohydrates constituted 15-50% of organic matter as compared with proteins (5-18%), amino acids (0.1-2.3%) and lipids (0.5-2.4%); high tide sediments values were higher than those of the low and mid tide sediments. Morphological examinations suggested that the microbial colonies are belonging to the genus Staphylococcus sp., Pseudomonas aeruginosa, Escherichia coli, Klebsiella sp., Bacillus sp., etc. The mining of placer mineral rich zone contributes to a mixed effect on bacterial and biochemical variables of the sediments. It has a negative impact on bacterial abundance and viability but positive impact on culturability. Viability and culturability could act as important indicators reflecting the disturbance in the system at short time intervals. Restoration of mining site can be done based on the results of culture test carried out in the laboratory. This baseline information about the potential mining site could help in developing rational approach towards sustainable harnessing of resources with minimum damage to the ecosystem.
*Corresponding author, E-mail address: vkhridya@gmail.com 2014 Journal of Coastal Sciences. All rights reserved

1. Introduction
The oceans cover more than 70% of the earths surface and little is known about the microbial diversity of marine sediments, which is an inexhaustible resource that has not been properly exploited. However, the full potential of this domain as the basis for biotechnology, particularly in India, remains largely unexplored. India with a long coastal line of over 7,500 km, an area of 2.02 million sq km in our exclusive economic zone, with very rich biodiversity, gives us an opportunity to investigate the mankind and ultimately for the economic uplift of India. The Tamil Nadu coastal region has diverse marine habitats such as seashore, hyper saline lakes, estuaries, saltpans and a variety of soil habitats. It is well known that sandy beaches form a major component of the coastal ecosystem and dominate temperate as well as tropical coastlines (Davies 1972). The long stressful abiotic factors such as the winds and waves make these beaches highly dynamic and ecologically sensitive (Brown and McLachlan 1990; Knox 2001). This stress is further compounded by human activities such as tourism and mining. Studies have shown that the sand mining activity causes an imbalance in the physical structure of the beach (Defeo et al. 2009). The most obvious effect of any type of mining is often the disturbance or displacement of large quantities of sediment, which in turn affects landforms and coastal processes (Hilton and Hesp 1996). Ecologically, the flora and fauna may also be affected (Lee and Correa 2005; Simmons 2005). However, little is known about the effect of mechanical disturbance on ecosystems especially at the microbial and biochemical levels in placer-rich-beach-sediment. Organic matter in marine sediments is derived from terrestrial and marine
27

sources (Premuzic et al. 1982; Gagosian and Peltzer 1986). Since aquatic organisms like algae are more abundant in the marine environment, primary productivity is probably an important factor controlling the distribution of organic molecules in sediments (Deuser and Ross 1980; Betzer et al. 1984). Prokaryotic organisms (Bacteria and Archaea) that live in marine sediments participate in a variety of biochemical pathways involving both inorganic and organic compounds. Analyses of the microbial community composition of many types of sediment in different parts of the world have shown that the predominant phylogenetic groups in these habitats are often highly similar. Placer deposits are formed as a result of the selective concentration of valuable minerals from the disintegration and redeposit ion of the rock fragments. Beach sands contain the most economically important minerals accumulations; wave action deposits sand on the beach and the heavy minerals are concentrated when backwash carries some of the lighter minerals such as quartz back into the sea. Heavy minerals have important economic applications. Their use in paleo-geographic reconstructions, especially in elucidating sediment transport pathways, is of particular value in hydrocarbon exploration, and their use in correlation has important applications in hydrocarbon reservoir evaluation and production. Recent advances have made it possible to utilize the technique on a real-time basis at the well site, where it is used to help steer high angle wells within the most productive reservoir horizons. Heavy minerals may become concentrated naturally by hydrodynamic sorting, usually in shallow marine or
ORIGINAL ARTICLE

J O U R N A L

O F

C O A S T A L

S C I E N C E S

fluvial depositional settings. Naturally occurring concentrates of economically valuable minerals are known as placers, and such deposits have considerable commercial significance. Cassiterite, gold, diamonds, chromite, monazite, and rutile are among the minerals that are widely exploited from placer deposits. Recently heavy mineral studies widely used in distinguish the tsunami sediment deposits from modern beach sediments. The influence on bacterial community and biochemical variables through mechanical disturbance of sediments is the case to undertake this problem in a placer rich zone of southern coast of Tamilnadu. These studies have usually been ignored to understand the environmental issues in beach mining in India. Therefore, in the present study, placer enriched zone, Kanyakumari to Inayam on the coastal belt of low, mid and high tide regions of southern coast Tamil Nadu is selected to study the bacterial dynamics for the sustainable development and harvesting the resources.

3. Methodology
3.1. Sample collection Samples were collected from 10 stations using along the coast between Kanyakumari and Inayam during July 2012 by core sampling method. Sediments have been collected from low tide, mid tide and high tide areas. The heavy mineral distribution is keenly observed on the mode of distribution like disseminated, laminated and change in colour of the sand due to variation in the type of materials segregated. The total of 30 samples has been collected in the study area. 3.2. Heavy mineral separation 3.2.1. Pre-treatment of sediments The samples collected from stations were heated at 100C on overnight in hot air oven for removing moisture content. Weighed approximately 100g after repeated coning and quartering process (for homogenous distribution) of sediments. The samples were washed and decanted with distilled water several times and kept for drying. Treated with 30% H2O2 and a pinch of SnCl2 to remove the organic debris and treated with 1:1 HCl after drying to remove calcareous materials present in sediments. 3.2.2. Sieving and Heavy mineral separation Sieving was carried out in ASTM sieves at 1/4 intervals. The sieve sets, stacked in descending order of size, were shaken using a Ro-tap sieve shaker continuously for about 20 min. During sieving, proper attention was paid to minimize the sand loss from the sieve sets. The sieved materials were collected separately for weighing (dry weight) and kept the data for further studies. Then the sieved fractions of each station of various tides were mixed together and separated in to heavy mineral fractions with bromoform of specific gravity 2.89. Separated fractions were washed with methyl alcohol

2. Study area
The study was conducted from Kanyakumari to Inayam (Kanyakumari coastal belt) in the south coast of Tamil Nadu located between 8 05 30 to 80 34 30 N and 770 06 30 to 770 35 00 E (Fig 1). The basement of this study area consist of charnockite, granite gneiss, leptynite, leptynite gneiss, peninsular gneiss, laterite, Warkalai sand stone, variegated clay, sandy clay, river alluvium etc. Alluvium occurs as the upper layer and is characterized by sand, gravel and sandy clay. The alluvium and weathered crystalline charnockites function as an unconfined aquifer system. The geomorphic and land use patterns existing in the study area are the beach sand, pediments, sand dunes, beach terraces, mud flats, saline tracts, water bodies etc. Sand dunes form a characteristic geomorphic feature of the study area and exist as a narrow band of approximately about 100m width along the topographically high areas.
Fig. 1 Location map of the Study Area

28

ORIGINAL

ARTICLE

J O U R N A L

O F

C O A S T A L

S C I E N C E S

followed by distilled water. Dried fractions were weighed for 4. Results and discussion calculating average weight percentage of placer deposits in each station. India's coastline of 6800 km is hosted by placer deposits of various grades and size. Indian resources constitute about 35 % of world 3.3. Biochemical composition of sediments resources of ilmenite, 10 % of rutile, 14 % of zircon and 71.4 % of monazite (Rajamanickam et al. 2005). Loveson et al. (2008) observed 3.3.1. Carbohydrate the inferences from sudden changes in the sedimentological processes during the December 26, 2004 tsunami along the east Total carbohydrate estimation were analysed by Phenol-Sulphuric coast of India. In the present study, out of 10 selected stations, acid method reported by Kochert (1978) based on a procedure sediments collected from high tide regions showing high percentage developed by Dubois et al. (1956). 1mL distilled water, 2mL phenol of placers. The enrichment in high tide and its reduction in mid tide (5% w/v) and 5mL concentrated H2SO4 were added to known and low tide are accounted by oscillations as well as by differing amount of sediment samples. D-Glucose was used as the standard energy conditions in the segregation of heavy minerals. However, and absorption was measured at 490nm. Kanyakumari (KK) beach samples suggest the withdrawal of sea level to low tide zone from the existing high tide zone (Fig 2, 3 & 4). This is 3.3.2. Protein indicated by the presence of very low percentage of heavy minerals Proteins were analyzed by Lowrys method explained by Herbert et al. (1971). A weighed amount of sediment sample is homogenized in 1N NaOH, and heated for 30 minutes at 80C. After cooling 1mL aliquots were added to a test tube containing carbonate-coppertartrate solution, followed by 0.5mL Folin-Ciocalteu reagent. Bovine Serum Albumin was used as the standard, and absorption was measured at 660nm. 3.3.3. Amino acid Amino acid estimated with Ninhydrin Method by adding 80% ethanol to a soil suspension and centrifuged at 15,000rpm. The collected supernatant diluted with distilled water followed by 1mL Ninhydrin reagent and boiled. Glycine was used as the standard at 540nm 3.3.4. Lipids Lipids were extracted from freeze-dried sediment samples by direct elution with chloroform and methanol (5:10). Extract was dried under vacuum (Bligh et al. 1959) and oxidized with 0.15% acid dichromate (Parsons et al. 1984). Stearic acid was used as a standard and absorbance was measured at 440 nm. 3.4. Microbial abundance and culturability The samples were stored in aseptic condition is cultivated on Zobell Marine agar (Peptone 5.0g, Yeast Extract 1.0g, Ferric Citrate 0.1g, Sodium Chloride 19.45g, Magnesium Chloride 8.8g, Sodium Sulfate 3.24g, Calcium Chloride 1.8g, Potassium Chloride 0.55g, Sodium Bicarbonate 0.16g, Potassium Bromide 0.08g, Strontium Chloride 34.0mg, Boric Acid 22.0mg, Sodium Silicate 4.0mg, Sodium Fluoride 2.4mg, Ammonium Nitrate1.6mg, Disodium Phosphate 8.0mg, Agar 15.0g). The colonies obtained were individually streaked into different petriplates and identified biochemically and macroscopically. Visible colonies present on the agar surface were counted using Neubauer Colony Counter and the counted colonies were expressed as colony forming units (CFU) per gram of Soil. Each colony was identified separately. Colony forming units No. of colonies X dilution factor Volume of sample added
Fig. 2 Placer deposits in low-tide marine sediments

Fig. 3 Placer deposits in mid-tide marine sediments

in lower layer. Manakudy (MK), Pallam (PL), Sankuthurai (ST), Muttam (MU) and Midalalm (MI) stations are also placer enriched zones. Midalam is a potential mining site for placers. Rajakkamangalam (RA), Kadiapattinam (KA), Colachel (CO) and Inayam (IN) stations are not much rich in placer deposits. Scientific mining operations accompanied by ecological restoration and regeneration of mined wastelands and judicious use of geological resources, with search for eco-friendly substitutes and alternatives must provide sensational revelation to the impact of mining on human ecosystem (Chauhan 2010).

29

ORIGINAL

ARTICLE

J O U R N A L

O F

C O A S T A L

S C I E N C E S

Fig. 4 Placer deposits in high-tide marine sediments

The biochemical composition of sediments such as amino acids, proteins, carbohydrates and lipids give the ratio of organic matter present in coastal mining sites. All the organic matter deposited in the sea is derived ultimately from some form of plant life, because, in the ocean, as on land, the animals depend directly or indirectly upon plants for food (Brandt 1900; Hensen 1887). The total carbohydrate percentage varies from 15-50% in each station (Fig. 5).

Fig. 6 Percentage of aminoacid in marine sediments

Fig. 7 Percentage of proteins in marine sediments

Fig. 5 Percentage of carbohydrate in marine sediments

The percentage of carbohydrate in low tide sediments shows high value compared to mid regions. This suggests that the acid and alkali soluble carbohydrates are either utilized preferentially in the water column during sedimentation (terrestrial or marine) or by benthic organisms. Acid- and alkali-soluble carbohydrates of particulate matter generally decrease with water depth (Hitchcock 1977). High tide value of carbohydrate is also high because of South West monsoon period. Vascular plants are known to contribute to sedimentary carbohydrates (Bhosle and Dhople 1988). Protein concentrations are high in all the stations compared to its free amino acids, and both high and low tide show a higher percentage than mid tide regions (Fig. 6 & 7). The presence of amino acid is probably due to the decomposition by microbial activity, and hydrolysis because of the redox conditions in the sediments or their adsorption to the sediments (Henrichs and Farrington 1987). The increased sedimentary protein suggests increased bacterial activity due to decay of other organisms. Lipid variation in different stations (Fig. 8) shows a similarity to proteins and carbohydrates.
30

Fig. 8 Percentage of lipids in marine sediments

The total biochemical composition in Kadiapattinam (KA) can clearly see as high, this may be due to the sewage effluent from human populated area. Kanyakumari (KK), a well known tourist place, shows the lowest biochemical composition for carbohydrate, protein, amino acids and lipids, because relatively little organic matter derived from land seems to be deposited in the sediments in the open ocean. And there may not have the chance for mixing fresh water nutrients from estuaries. Grain size of the sediments also plays a significant role in the distribution of biochemical components due
ORIGINAL ARTICLE

J O U R N A L

O F

C O A S T A L

S C I E N C E S

to hydrodynamic activities or mining. Seasonal variation of erosion and accretion processes are noticed along the entire study area except for Midalam and Inayam where the hydrodynamic activities are interfered by revetments, sea walls and groins (Prince et al. 2014). The sediments in the open ocean, on the other hand, are

Fig. 9 Microbial colonies in low, mid & high tides Stations Kanyakumari Manakudy Pallam Sankuthurai Rajakkamangalam Muttam Kadiapattinam Colachel Midalam Inayam low-tide Coliforms, Bacillus, Staphylococci Bacillus, E.cloi, Klebsiella Coliforms, Bacillus Staphylococci, Bacillus, E.cloi Bacillus, E.cloi, Pseudomonas Bacillus, E.coli, Klebsiella Coliforms, Aspergillus, Actinobacter Staphylococci Coliforms, Bacillus, Staphylococci Bacillus, E.cloi

The most obvious effect of any type of mining is often the disturbance or displacement of large quantities of sediment which in turn affects landforms and coastal processes (Hilton and Hesp 1996). Thus, any mechanical stress in the ecosystem is reflected in the distribution pattern of this parameter and its relationship with other physico-chemical and biological parameters. The potential mining site Midalam shows better cell abundance and culturability (Fig. 9) and its biochemical components are not much lesser compared to other stations. Examination of interrelationships between bacterial and biochemical parameters suggested the extent of interdependence between them. The high tide region of Kadiapattinam station shows a higher value of culturability than all other stations. The biochemical composition of this area in its low tide was the highest. The placer rich zones and other stations dont show any exhibited variations in both bacter ial and biochemical properties. The common marine microbial colonies are belonging to the genus Staphylococcus sp., Pseudomonas aeruginosa, Escherichia coli, Klebsiella sp., Bacillus sp., etc. can see in both placer rich and unrich zones of the study areas (Table 1). Christabelle et al. (2012) has studied before and after effect of a particular mining site, and reported as the mechanical disturbances could affect short term life span of bacterial community. This could clearly see in both the biochemical bacteriological composition of beach sediments in placer rich zones. Thus, the study in a sand mining beach ecosystem assesses the short-term effect on bacterial and biochemical parameters over short term periods. While total counts and viability decreased, culturability increased and improved the liability of the available substrates.
High-tide Coliforms, Bacillus, Aspergillus Staphylococci, Bacillus, E.cloi E.cloi, Pseudomonas Coliforms, Staph.albus Bacillus, Coliforms, Staph.albus Aspergillus, Staph.albus Staphylococci, Klebsiella, E.coli,Pseudomonas Pseudomonas, Klebsiella Bacillus, E.cloi, Pseudomonas Pseudomonas, Actinobacter

Mid-tide Bacillus, E.cloi, Staphylococci Bacillus, E.cloi Bacillus, Staphylococci Pseudomonas, Klebsiella Bacillus, E.cloi, Staphylococci, Klebsiella Pseudomonas, E.cloi, Klebsiella Bacillus, E cloi Bacillus, Pseudomonas Bacillus, E.cloi Bacillus, E.cloi

Table. 1 Bacterial abundance in various stations of study area

deposited, according to Schott (1933), at a rate of about 1cm in 1000 years and they contain about 1% organic matter. This means that 0.2g of organic matter would accumulate on a square meter of sea bottom each year, or 0.02% of the original supply of plankton in the water above. Thus, when the rate of deposition of marine sediments is rapid, about one-fiftieth of the original organic matter seems to be deposited, and, when it is slow, about one-five thousandth accumulates in the sediments. The small proportion that seems to be laid down in sediments that are deposited very slowly may be due to better chances for oxidation or destruction of the organic matter before it is buried effectively. Thus the mining periods gap should be higher for conserving the biochemical compounds effectively (Christabelle et al. 2012). Bacterial community also drives the spatial distribution metabolism and dynamics of most benthic organisms. Further this community is involved in prime utilisation and net mineralisation of sedimentary liable organic matter which is primarily composed of carbohydrates, lipids and proteins.
31

5. Conclusions
The selected study area has both placer rich and poor stations with negligible variation in its distribution of both biochemical and bacteriological data. The mechanical disturbances like mining activity improved the culturability of bacteria, though it may have a negative impact on the total bacterial abundance. Spatio-temporal observations along the mining stretch can provide a better outlook to the biochemical factors responsible for the existence of microbial community. The influence of active sand mining on a larger microbial community and its long-term impact can be studied further on sedimentary microbes other than bacteria like the protozoan on a longer time scale.

References
Betzer, P.R., Showers, W.J., Laws, E.A., Winn, C.D., Diiullio, G.R., Kroopnick, P.M., 1984. Primary productivity and particle fluxes on a transect of the equator at 153W in the Pacific Ocean. Deep-Sea Research 31, 1-11. ORIGINAL ARTICLE

J O U R N A L

O F

C O A S T A L

S C I E N C E S

Bhosle, N.B., Dhople, V.M., 1988. Distribution of some biochemical compounds submitted to Dept. of Ocean Development, Govt. of India, New Delhi, 220 in the sediments of the Bay of Bengal. Chemical Geology 67, 341352. pp. Bligh, E.G., Dyer, W.J., 1959. Rapid method of total lipid extraction and Schott, W., 1933. Die jungste Vergangenheit des quatorialen Atlantischen purification. Canadian Journal of Biochemistry and Physiology 37(8), 911Ozeans auf Grund von Untersuchungen an Bodenproben der 'Meteor' 917. Expeditions," Sitz. Ber, u. Abhandl. Naturforsch. Gesell. Rostock, Dritte Brandt, K., 1900. Beitrge zur Kenntniss der chemischen Zusammensetzung Folge, Bd. 4, pp. 48-59. des Plankton, Wiss. Meeresunt., Kiel, N. F., Bd. 3, pp. 43-90. Simmons, R.E., 2005. Declining coastal avifauna at a diamond mining site in Brown, A.C., McLachlan, A., 1990. Ecology of Sandy Shores. Elsevier, Namibia: comparisons and causes. Ostrich 76, 97103. Amsterdam, Netherlands. Chauhan, S.S., 2010. Mining, Development and Environment: A Case Study of Bijolia Mining Area in Rajasthan, India. Journal of Human Ecology 31(1), 65-72. Christabelle, E.G.F., Anindita Das, Nath, B.N., Daphne G.F., Loka Bharathi, P.A., 2012. Mixed Response in Bacterial and Biochemical Variables to Simulated Sand Mining in PlacerRich-Beach-Sediments, Ratnagiri, West Coast of India. Environmental Monitoring and Assessment 184, 2677-2689. Davies, J.L., 1972. Geographic variation in coastal development. Longmans, London. Defeo, O., McLachlan, A., Schoeman D.S., Schlacher T.A., Dugan, J., Jones, A., Lastra, M., Scapini, F., 2009. Threats to sandy beach ecosystems: A review. Estuarine, Coastal and Shelf Science 81, 1-12. Deuser, W.G., Ross, E.H., 1980. Seasonal changes in the flux of organic carbon to the deep Sargasso Sea. Nature, 283, 364-365. Dubois, M., Gillies, K.A., Hamilton, J.K., Reebers, P.A., Smith, F., 1956. Calorimetric method for determination of sugars. Analytical Chemistry 28, 350-356. Gagosian, R.B., Peltzer, E.T., 1986. The importance of atmospheric input of terrestrial organic material to deep sea sediments. Organic Geochemistry 10, 661-669. Henrichs, S.M., Farrington, J.W., 1987. Early diagenesis of amino acids and organic matter in two coastal marine sediments. Geochimica et Cosmochimica Acta 51, 1-15. Hensen, V., 1887. ber die Bestimmung des Planktons oder desim Meere treibenden Materials an Pflanzen und Tieren, Kommission zur wiss. Untersuchung der deutschen Meere, in Kiel, 1882-1886, Bericht , Vols. 1216, pp. I-107. Herbert, D., Phipps, P.J., Strange, R.E., 1971. Chemical analysis of microbial cells. In: J.R. Norries and D.W. Ribbons (Editors), Methods in Microbiology, Vol. 5B. Academic Press, London, pp. 244-252. Hilton, M.J., Hesp, P., 1996. Determining the limits of beach-nearshore sand systems and the impact of offshore coastal sand mining. Journal of Coastal Research 12(2), 496519. Hitchcock, G.L., 1977. The concentration of particulate carbohydrate in a region of the West Africa upwelling zone during March 1974. Deep-Sea Research 24, 83-93. Knox, G.A., 2001. Soft shores. In: The Ecology of seashores. Kennish, M.J. (ed.). CRC Press, LLC, Boca Raton. Kochert, G., 1978. Carbohydrate determination by phenol-sulfuric acid method. In:Hellebust, J. A., Craige, J. S. (eds.), Handbook Of Phycological Methods - hysiological and Biochemical methods. Cambridge University Press, Cambridge. Lee, M.R., Correa, J.A., 2005. Effects of copper mine tailings disposal on littoral meiofaunal assemblages in the Atacama region of northern Chile. Marine Environmental Research 59, 118. Loveson, V.J., Angusamy, N., Gujar, A.R., Chandrasekar, N., Rajamanickam, G.V., 2008. Observed inferences from sudden changes in the sedimentological processes during the December 26, 2004 tsunami along the east coast of India. Science of Tsunami Hazards, 27(4), 43 52. Parsons, T.R., Maita, Y., Lalli, C.H., 1984. A Manual of Chemical and Biological Methods for Seawater Analysis. Pergamon, Oxford. Premuzic, E.E., Benkovitz, C.M., Gaffney, J.S. and Walsh, J.J., 1982. The nature and distribution of organic matter in the surface sediments of world oceans and seas. Org. Geochem., 4: 63-72. Prince S. Godson, Chandrasekar, N., Joe Vivek, V., Krishna Kumar, S., 2014. Seasonal variability in sediment distribution along the southwest coast of Tamil Nadu, India, International Journal of Recent Scientific Research 5(1), 44-53. Rajamanickam, G.V., Angusamy, N., 2005. Exploration of placer deposits between Vedaranyam and Pondicherry, Tamil Nadu, India, Technical report 32

ORIGINAL

ARTICLE