Predominantly Occurring Phytoplankton in Ariyankuppam Coastal Waters, Southeast Coast of India

INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 9, ISSUE 03, MARCH 2020 ISSN 2277-8616 Predominantly Occurring Phytoplankton In Ariyankuppam Coastal Waters, Southeast Coast Of India

M. Punithavalli, K. Sivakumar

Abstract: The study were conducted for six months covering summer and pre monsoon seasons to analyze the seasonal variation on phytoplankton in relation with hydrological parameters in Ariyankuppam coastal waters, south east coast of India. The physico-chemical parameters such as atmospheric temperature from 32.16 to 34.23°C, water temperature from 31.13 to 33°C, pH from 8.0 to 8.3, salinity from 29.33 to 33.66‰, dissolved oxygen 3.7 to 4.06 mg/l and nitrate 0.06 to 0.095(mg/l). A total of 25 taxa were recorded dominated by Bacillariophyceae (19) followed by Dinophyceae (5) and Cyanophyceae (1). Further predominantly occurring marine phytoplankton were Coscinodiscus radiatus, Odontella mobiliensis, Navicula sp, Thalassiosira sp, Triceratium sp, Pluerosigma sp, Skeletonema sp, Ceratium furca, Ceratium sp, Dinophysis tripos and Protoperidinium depressum. Commonly occurred genera, Chaetoceros (Chaetocerotaceae), Coscinodiscus (Coscinodiscaceae) and Navicula (Naviculaceae), were subjected to Energy Dispersive Spectroscopic analysis (EDS). They were found to accumulate different, element such as Na, Mg, Si, Cl, K, Cu, Zn, Cr and Fe. Among these the member Chaetoceros contained Na, Mg, Si, Cl, K, Cu and Zn, Coscinodiscus Na, Mg, Si, Cl, Cu, Zn and Navicula Mg, Si, Cl, K, Cu, Zn, Cr and Fe. Thus these observations would determine the chemical dialogue between the cell structures and role of the elements. Further, it gives the clue about the phytoplankton growth requirements.

Keywords: Ariyankuppam, Physico-chemical parameters, Phytoplankton, SEM-EDS ————————————————————

1 INTRODUCTION The concept of employing plankton as indicator species and Phytoplankton act as important component of the marine species diversity is also the basis for functional compensation, ecosystem, liberate oxygen during photosynthesis and aid in which may leads to the effect of biodiversity on ecosystem exchange process [14] and are very sensitive to change in its functioning through compensatory dynamics. The species environment before it becomes visible on higher tropic levels and present in a community have functional roles, this may be excessive they live in any alteration in the environment leads to expected to increase total production and also include their the change in the plankton communities in terms of tolerance, abundance and biomass or growth (blooms) of algal species abundance, diversity and dominance in the habitat [6]. They play because of anthropogenic activities [1]. But still there are many a major role in global warming, by reducing the global corban such aquatic ecosystem that remains unexplored. The dioxide level [26]. The relationship between phytoplankton and Ariyankuppam seashore is one among them which has not nutrient is highly dynamics and always been the major focus received due attention. Hence, the present work is an attempt to among the explicate experimental ecology [8]. The various study the influence of physico-chemical parameters of water on anthropogenic activities are increased, which in turn to enhance phytoplankton population, species composition and community the nutrient concentration and the leads to high productivity in organization. The chemical composition of phytoplankton coastal environment [22]. The investigations of hydrological sample used for their calibration has been currently features are necessary for assessing the fertility and productivity determines by bulk analysis technique [5]. This leads to of any ecosystems [21]. The physico-chemical parameters estimation of elemental concentration for the whole samples quantity of nutrients studies of phytoplankton have been utilized without separation into separate constituents eg., algae, to assess the diversity, ecology and quantity of water of the detritus, small animal, etc., inevitably such data on natural plankton [20, 31]. Coastal ecosystem is highly vulnerable to bloom of various phytoplankton species could therefore pollution by anthropogenic activities. The human interventions, contain errors due to the presence of other species and or particularly increasing population, industrialization and tourism in detritus particles in the analyses samples. In the study, the the coastal areas are the major threats to the coastal marine elemental composition of phytoplankton is determines for environment. Such activities ultimately bring eutrophication in the individual species by X-ray micro analysis (XRMA), Thus marine environment and consequently into discrepancy of marine avoiding the above problems, XRMA has sufficient resolution ecosystem. Phytoplankton dynamics is effectively controlled by to analyze single phytoplankton cells with a mixed population few physical factors along with hydrographic properties and thus [9] and involves the simultaneous determination of a range of exhibit the seasonality pattern. elemental concentration for each cell-thus providing a comprehensive assessment of nutrient status. [27] This research represent first attempt to examine an microalgae with SEM-EDS.

2 MATERIALS AND METHODS ______

• M. Punithavalli, Ph.D. Research Scholar, Department of Botany, Phytoplankton analysis Annamalai University, Annamalai Nagar - 608 002, India. E-mail: The water samples were collected from Ariyankuppam [email protected] [latitude 1188’05.08‖N, longitude 7982’59.87‖E] coastal waters • K. Sivakumar, Division of Algal Biotechnology, Department of at monthly intervals for a period of six months from April to Botany, Annamalai University, Annamalai Nagar - 608 002, India. E-mail: kshivam69@gmail. September 2018 for the estimation of temperature, pH, salinity, dissolved oxygen and nitrate. Phytoplankton samples 6560 IJSTR©2020 www.ijstr.org INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 9, ISSUE 03, MARCH 2020 ISSN 2277-8616 were collected by plankton net (mesh size 20 μm) made up of The physico chemical factors and biological factors showed bolting silk cloth. The samples were collected in black variation between monthly during the six months study period. polythene bottles and immediately preserved with 4% formalin The physico-chemical parameters at Ariyankuppam coastal for quantitative and qualitative analysis. Some of the samples waters during the study period of April to September 2018 are were fixed in 3% glutaroldehyde in 0.1 M phosphate buffer (pH presented in Table.2. Among the physico-chemical 7.0) for scanning electron microscopic studies. Samples were parameters variations is temperature is one of the most then dehydrated through a graded series of alcohol 12-15 min important factors that have an influence on the distribution in interval at 4˚C. Then the dehydrated phytoplankton samples the marine ecosystems. In the present study the highest treated with critical point drier (CPD) were kept on a stub and the atmospheric temperature recorded during summer season samples were coated with gold palatium and examined in Joel 34.23˚C in the month of May and the lowest temperature JSM-56010 LV with SEM photomicrographs were taken 32.16˚C in the month of September Premonsoon season. selectively from the computer screen. Phytoplankton were Water temperature recorded during summer season 33˚C in identified using the previous works of [37, 36, 10, 32, 25, 2]. the month of May and lowest temperature 31.13˚C during Simultaneously selected portion of micrograph was subjected premonsoon season in the month of September. The to Scanning Electron Microscopy (SEM) and Energy observed high value of temperature in May was due to the Dispersive Spectroscopic analysis (EDS). This was conducted intensity of solar radiation and evaporation freshwater influx with an EDS 700 series interfaced with a data general NOVA2 and cooling and mix up with ebb and flow from adjoining computer and a Texas instrument silent 700 ASR. The EDS X- neritic waters [34]. The observed low temperature in the month ray spectrometer was interfaces with a scanning electron of September was due to strong land sea breeze and microscope (20 kv) stage. The area of different components precipitation [30]. Salinity is one of the great significant factors to such as cell wall and cellular inclusion was analyzed. To find marine life in general that regulates the quality of waters and out the fluxes of particular mineral, both the counts per second determines the extent of dissolution of gases and hydrogen ion (S-1 or CPS) value and the apparent relative atomic concentration. In the present study the highest salinity of 33.66 percentage of weight in different components of the cell wall (‰) was recorded in the month of May summer season which and cellular inclusion details were documented. can be attributed to low rainfall, decreased freshwater inflow, elevated atmospheric temperature and evaporation [23]. The lowest salinity of 29.33 (‰) was recorded in the month of September Premonsoon season may be due to heavy rainfall and large quantity of freshwater inflow as reported by [4, 33]. Salinity act as limiting factors in the distribution of living organisms and its variation caused by dilution and evaporation is most likely to influence the fauna in the coastal ecosystems [35]. The hydrogen ion concentration plays on important role in many of the life processes of aquatic organisms. In the present study minimum pH value of 8.0 recorded during the premonsoon season in the month of September may be due to the freshwater influx and subsequent dilution of seawaters, reduction of temperature, reduction of salinity and decomposition of organic matter [19]. The maximum values recorded during summer season 8.3 in the month of May may be due to the uptake of CO2 by photosynthesizing organisms etc., [12, 24]. The dissolved oxygen is a major component of aquatic ecosystem and

quality of water support aquatic life. In the present study the Figure 1: Map showing the study area high oxygen concentration was observed during premonsoon 4.06 (mg/l) in the month of September increase in oxygen concentration may be due to the influx of oxygen 3 RESULTS AND DISCUSSION concentration rich freshwater in to the coastal waters and A total of 25 phytoplankton species were recorded in the present cumulative effect of higher wind velocity coupled with heavy study from the water sampling stations of the Ariyankuppam rainfall. The increase in the phytoplankton density coast. They belong to three groups such as bacillariophyceae premonsoon contributed to the increase in oxygen (diatom), dinophyceae (dinoflagellates) and cyanophyceae (blue concentration [16]. Lowest oxygen concentration was green algae). The phytoplankton diversity was higher in May observed during summer season 3.53 (mg/l) in the month of (Table.1). The commonly occurring species were Amphora May. Nitrate is thermodynamically the most stable from the proteus, Cosinodiscus radiatus, Leptocylindrus danicus, Navicula combined inorganic nitrogen in well oxygenated waters. The sp, Nitzschia longissima, Rhizosolenia imbricata, Skeletonema maximum concentration of nitrate recorded during premonsoon costatum, Thalassiosira eccentrica and Trichodesmium thiebautii. seasons in the month of September. The recorded high nitrate Diatoms formed the most dominant group (76%) followed by values could be attributed to the increased phytoplankton dinoflagellates (20%) and blue green algae (4%); however, a excretion, oxidation and reduction by recycling of nitrogen and visible change was noticed during May the bloom of also due to bacterial decompression of planktonic detritus present Trichodesmium thiebautii occurred. The diatoms were the in the environment. Similar observation was reported by [13] in dominant phytoplankton prevailing in the coastal waters of the present study. The minimum concentration of nitrate Ariyankuppam, covering the summer and premonsoon periods. recorded during summer

Table 1: List of phytoplankton species at Ariyankuppam season 0.06 (mg/l) in the month of May be due to its utilization by coastal water during the Period of April to September 2018 phytoplankton as evidenced by high photosynthetic activity and Species name April May June July August September also neritic waters dominance, which contained only negligible Bacillariophyceae amount of nitrate. Seasonal variation of phytoplankton the high Amphora proteus + + + + - - abundance of phytoplankton starting from monsoon and (Gregory 1857) Asterionellopsis extending to summer months in the present study may be due glacialis (Round + + + + + + to the influence of pH, Salinity, temperature, elevated nutrient 1990) concentration and high level of light Penetration similar Bellerocha malleus + + + - + - observation was also made [17, 3]. The density of phytoplankton (Van Heurck 1885) increased considerably and reached the maximum during Bacillaria paradoxa - + + + - + summer and minimum during premonsoon [18]. The dominance (J.F. Gmelin 1791) Cyclotella sp. (A. de of diatoms during summer seasons has also been reported in - + - + - - Brébisson 1838) previous studies from the east coast of India [11]. The members Coscinodiscus of Bacillariophyceae were dominant among the diatoms during radiatus (Ehrenberg + + + + + + the present study. The dominant diatoms recorded during the 1840) study period in Asterionellopsis, Bellerocha, Coscinodiscus, Chaetoceros capense (Karsten, + + + + - - Chaetoceros, Leptocylindrus, Navicula, Nitzschia, Pleurosigma, 1905) Rhizosolenia, Skeletonema were observed. The present Ditylum brightwelli investigation on the hydrographic variations and abundance of + - + + + - (Grunow 1885) diatoms provides valuable and add a list of information on the Fragilaria sp. (H.C. + + - + + - seasonal variation of phytoplankton population in relation to Lyngbye 1819) Guinardia flaccida various environmental factors. - + - + - - (H. Peragallo 1892) Leptocylindrus Correlation matrix of physico-chemical parameters danicus (Cleve + + + - + + The correlation coefficient values of different physico chemical 1889) parameters data at Ariyankuppam coastal waters samples are Navicula sp. (J.B. presented in Table.2. The physico chemical parameters M.Bory de Saint- + + + + + + vincent 1822) showed a positive correlation with atmospheric temperature, Nitzschia water temperature, pH and salinity is measured because of its longissima + + + + + + influence of the distribution and diversity of many living marine (Brébisson) (Ralfs species and a negative correlation with dissolved oxygen and 1861) nitrate. Odontella mobiliensis - + - + + - (Grunow 1884) Description of the species Pleurosigma sp. (W. Scanning Electron Microscopic observation made on the two + + + + + + Smith 1852) different classes (Fig.2 and 3). Rhizosolenia imbricata (Brightwell + + + - + - 1858) a) Coscinodiscus radiates (Ehrenberg 1840): Coscinodiscus Skeletonema radiatus in the size of the marginal microrimoportulae, smaller costatum (Greville) + + + + + + and almost indistinguishable from the microrimoportulae and (Cleve 1873) large and easily distinguishable from the microrimoportulae in Triceratium favus - + + + - + Coscinodiscus radiatus in the morphology of the (Ehrenberg 1839) microrimoportulae long thick neck, trilobulate at the top and Thalassionema sp(Grunow ex C curved toward the mantle in Coscinodiscus radiatus the cribra - + + + + - Mereschkowsky pattern and morphology of the macrorimoportulae and 1902) microrimoportulae characteristic of Coscinodiscus radiatus valve Dinophyceae of small specimen without rosette. Arrowheads show scattered Ceratium furca rimoportula, valve of a large specimen with conspicuous rosette, (Ehrenberg) + + + + - + radial aredation (Fig.2.a). (Lachmann 1859) Ceratium lineatum (Ehrenberg) (Cleve - + + - + - b) Odontella mobiliensis (Grunow 1884) 1899) Cells single or rarely united in short straight chains by the long Dinophysis tripos + + + - + - spines. Two long spines are placed far apart, but about (Gourret 1883) equally far from the processes, directed obliquely outward, Protoperidinium depressum (Bailey + + + + + + straight or often bent abruptly in their outer part. Cells 1854; Balech 1974) relatively thin-walled, without a sharp constriction between Prorocentrum valve and girdle zone. Sculpturing fine, reticulate, 14-16 areolae micans (Ehrenberg + + + + - - in 10 µm of the frustules focusing of show of convex, with flat 1834) frustules on valve and valve mantle, 17-18 on girdle band. Cyanophyceae Auxospores formed as large bladders from the separation of the Trichodesmium thiebautii (Gomont + + + + - + valves, with much larger cells inside (Fig. 2.b). 1890) c) Navicula sp. (Bory de Saint-vincent 1822)

Navicula species were abundant, occurring as solitary or two flagella, chloroplast numerous yellow brown, Mixotrophic, colonial forms, enclosed in mucilaginuous plates. sexual and asexual, bloom due to coastal eutrophication, cause Naviculaceae differs from Achanthaceae and toxic red tides, coastal, estuaries and ocean. This species has a Phaeodactylaceae by being isovalvar; both valves of a cell straight body which is 70-200 µm long and 30-50 µm wide, have a ―naviculoid‖ raphe not subtrended by the fibulae with in the epitheca gradually tapering into an anterior horn present in Bacillariophyceae. Many of the few marine planktonic (Fig.3.i). Navicula species were transferred to other genera, especially after Cox (1979) typified and emended the description of j) Ceratium sp. (Schrank 1793) Navicula sensu strict (Fig.2.c). They are easily distingued from other types of dinoflagellates because of their morphological characteristics, described d) Pleurosigma sp. (Smith1852) below. They are a relatively harmless group of organisms that Solitary pennate diatom with a gentle sigmoid shape. play an important role as both predators and prey in their Pleurosigma sp were observed crossing the slides. Cell size environment. The epitheca tapering into apical horn and the width 28-75 cm, length 90-600 cm. Valves are broadly antapicals are strong, unequal and straight; large, diverse lanceolate. Ends are blunt with sigmoid raphe. Length and genus. Around, gonyaulacoid body, two to four hollow horns. breadth of the cell vary between respectively (Fig. 2.d). Horns open or closed (Fig.3.j). e) Thalassiosira sp (Cleve 1873) k) Dinophysis tripos (Gourret 1883) single valve of Thalassiosira species with process pattern. Cell Species in this genus are laterally compressed with a small, in valve diameters is 19.47 µm and structure annulus show Cap-like epitheca and a much larger hypotheca (dorsoventral (arrow), scale 10 µm. Cells discoid to cylindrical solitary or depth of epitheca is 1/3 to 1/2 hypotheca). D. tripos is a very joined by treads or valve to valve to form loose; chains or in distinctive species. Cells are large, anterio-posteriorly mucilage masses. Plastids numerous discoid. Areolae usually elongated and asymmetrical with two posterior hypothecal loculate arrange in radial rows, tangential rows, or arcs; projections; a longer ventral process and a shorter dorsal one. varying size and prominence. Fultoportulae occur also on the The V shaped process are often toothed on their posterior valve face. 13-15 areolae in 10 µm. marginal strutted. ends (Small knob-like spines) (Fig.3.k). Scattered strutted process (arrow) and process pattern (circle). Copulae numerous spilt and ligulate. (Fig.2.e). l) Protoperidinium depressum (Balech 1974) Cells elongated, with rounded or quadrangular body with slightly f) Triceratium sp. (Ehrenberg 1839) convex or almost straight margins of both epitheca and Cells free living or attached; usually triangular in valve view and hypotheca. Ortho-quadra. Cingulum planozone, descending, narrowly oblong in girdle view; with elevation at the corners at a with about 1.5–2.0 cingulum width offset (depends much on slight central convexity. Valve face flat of slightly convex, mantles the angle of view). Apical horn is conical, not separated. The vary shallow, Areolae loculate, opening externally via large hypothecal pore is absent. Antapical horns are conical, the foramina; the bases of the loculesare formed by a continuous right one being slightly larger, ending in short strong divergent sheet of silica with rows of pores ladiating from a central annulus. spines. Length 132-165 µm, width 102-135 µm , height 78- Pores often eroded but in vivo with domed coverings; internally 102.5 µm. Many species have apical and antapical horns the pores are clustered but they still clearly radiate from the and/or antapical spines. Chloroplasts is almost absent (Fig.3. centre of the valve (see also Miller and Collier 1978). Corners l). elevations present, ending in ocelli. In some species there is a conspicuous rimoportula opening adjacent to each ocellus. Valve Table 2: Correlation matrix of Physico-chemical parameters mantle often with a ridge below which there is a fine row of pores variables at Ariyankuppam coastal waters April- September 2018 Dissol Nit just above the recuvred mantle edge. (Fig.2.f). Atmosph Water ved rat eric Tempe Salinit Parameters pH oxyge e h) Skeletonema sp. (Greville 1865) Tempera rature y (‰) n (m ture (°C) (°C) Cells joined by long marginal processes to form filaments, (mg/l) g/l) which appear in the light microscope like short beads joined by Atmospheric numerous fine threads. Plastids disc-like or cup-shaped. Living Temperature 1 in the coastal marine plankton valves circular, valve face (°C) convex to flat; mentles deep. A single ring of processes occurs Water around the top of the mantle. These are closely associated Temperature 0.984** 1 (°C) with a ring of fultoportulae, the external opening of which are pH 0.946** 0.906* 1 short tubes hidden in the bases of the processes. Occasional 0.955* Salinity (‰) 0.984** 0.932** 1 valves produce flattened spinulose processes and on these * Dissolved - - there is also a central rimoportula, with a tubular external -0.880* -0.849* 1 opening. Occasional rimoportulae also occur around the valve oxygen (mg/l) 0.906* 0.812* mantle, scale bar 20µm (Fig.3.h). Nitrate (mg/l) -0.617 -0.647 -0.583 -0.707 0.461 1 i) Ceratium furca (Lachmann 1859) Ceratium furca has a wide girdle and a prominent straight apical horn. C. furca also has two unequal posterior horns. The right horn is shooter than the left. There is a thin bar that connects. connection none (solitary), covering cellulose theca,

(Chaetocerotaceae) cell wall was 35% wt. compared to 27% wt of Mg. The Coscinodiscus (Coscinodiscaceae) six chemical elements in the following order: Si ˃ Cu ˃ Cl ˃ Zn ˃ Na ˃ Mg. contribution of Si being maximum 33% wt. Navicula (Naviculaceae) contained eight chemical elements in the following order : Cl ˃ K ˃ Cu ˃ Mg ˃ Si ˃ Zn ˃ Fe and Cr. Thus is constituent of different chemical elements in Chaetoceros, Coscinodiscus and Navicula varied not only by quality but also in quantity (Figure 4). The major element detected on phytoplankton cells correspond to these seen in other marine water algae, including the presence of Si. Thus element is generally regarded as being cell wall associated and has been detected by X-ray microalgae (XRMA) in a range of algae cells including blue green algae [7, 28, 15, 29]. In the present study the element Mg ˃ Si ˃ Cl ˃ Cu and Zn is commonly present in Chaetoceros, Coscinodiscus and Navicula. Among which the Chaetoceros seven chemical elements in the following order Cl ˃ Mg ˃Cu ˃ Zn ˃ Si ˃ Na ˃ K. Coscinodiscus Si ˃ Cu ˃ Cl ˃ Zn ˃ Na ˃ Mg and Navicula Cl ˃ K ˃ Cu ˃ Mg ˃ Si ˃ Zn ˃ Fe ˃ Cr.

Table 3: SEM-EDS (Energy Dispersive Spectroscopy) analysis of Chaetoceros, Coscinodiscus and Navicula

Chaetocerotacea Coscinodisca Naviculacea Name of the S.n e ceae e Chemical Figure 2: Scanning electron microscopic observation of o phytoplankton species elements Chaetoceros (wt. Coscinodiscu Navicula (wt. %) s (wt. %) %) 1 Na 4.08 6.15 - 2 Mg 27.23 4.11 12.64 3 Si 6.81 33.74 12.51 4 Cl 35.36 16.65 39.29 5 K 2.04 - 14.50 6 Cu 16.11 24.87 12.68 7 Zn 8.37 14.47 7.12 8 Cr - - 0.37 9 Fe - - 0.88 All the parameters are in triplicate values.

Figure 3: Scanning electron microscopic observation of phytoplankton species Figure 4: SEM-INSA-EDS analysis of Chaetoceros Table 3 shows the presence of different chemical elements in (Chaetocerotaceae), Coscinodiscus (Coscinodiscaceae) and the Chaetocerotaceae, Coscinodiscaceae and Naviculaceae Navicula (Naviculaceae) of marine water algae. The Chaetoceros (Chaetocerotaceae) contained seven elements in the following order: Cl ˃ Mg ˃ Cu The role of the chemical elements Sodium (Na) Sodium is a ˃ Zn ˃ Si ˃ Na ˃ K. The contribution of Cl in the Chaetoceros viable strategy to increase cellular abundance and 6564 IJSTR©2020 www.ijstr.org INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 9, ISSUE 03, MARCH 2020 ISSN 2277-8616 concentration of pigments and lipids in some microalgae as dynamic allow simultaneous consideration of a wide range of well as the rate of lipid accumulation in nitrate deplete chemicals. The approach presented here could be easily cultures. 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