1.

INTRODUCTION Dinoflagellates are one of the most important members of the phytoplankton in

marine and freshwater ecosystems which represent a major component in food webs (Saldiarriaga & Hoppenrath, 2010). Their cell sizes were range from 2 m to 20 m. More than 2,000 existing species have been described, only half of which are photosynthetic. They include autotrophs, mixotrophs and grazers (Taylor et al., 2007). Most of the photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate symbionts, plus all those crucial to reef-building corals (Taylor et al., 2007). Some of them are also the important sources of bioluminescence in the ocean which can light up at night (Hallegraeff, 1995). Beside diatoms, over half of the present-day dinoflagellates are photosynthetic and are at the base of marine food web (Spector, 1984). Furthermore, dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters, mussels, scallops and clams (Faust, 2002). Dinoflagellates can be identified by the presence of dimorphic flagella; one flagellum oriented around the cell, and the other directed posterior which allow them to swim freely in water with a forward spiralling motion (Faust, 2002). Another characteristic of the dinoflagellates is the wall configuration and arrangement; that was based on the presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known as theca. The dinoflagellates have unique characteristics. It contains a distinctive nucleus, named the dinokaryon. The DNA is not systematized around nucleosomal histones, as a substitute, it forming fibrillar chromosomes that are always condensed and that divide via a closed mitosis with an external spindle (Taylor et al., 2007). When dinoflagellate proliferates to higher than normal concentrations, the water may discolour and appears red. This condition is called red tides. The blooms may be

either toxic or noxious which they often accumulate in shellfish or fish, and when these are eaten by humans they cause diseases like paralytic shellfish poisoning (PSP), neurotoxic shellfish poisoning (NSP), diarrhetic shellfish poisoning (DSP) and ciguatera (Hallegraff, 2004). Besides human health, Harmful Algal Blooms (HABs) also give negative impact to aquaculture, tourism industries and to the ecology of the water system. Various factors have been associated to the occurrence of HABs phenomenon such as wind and currents, light, temperature, oxygen, pH and nutrient uptake (Burkholder et al, 2006). Some species were not bloom forming but have potential toxin producers. A few studies have done on dinoflagellate in Sarawak River, including Santubong River. The study provides information on the species that available at Sarawak River but the composition of dinoflagellates in the area of aquaculture is limited. The aquaculture effluent might have an influence to the species composition of dinoflagellates in that area and the potentially toxin producer will gives negative impact not only to the river system but also to the consumer who consume the seafood on that area. The objective of this study was to evaluate the occurrence of dinoflagellates species composition at aquaculture area along Santubong River. The composition of dinoflagellates in Santubong River were also identified and documented. This study could provide an understanding on the documentation and all taxa of dinoflagellates presence on the study area. More over, the possible toxic producers and bloom forming dinoflagellates can be identified.

2.0

LITERATURE REVIEW

Dinoflagellates are one of the most important components of phytoplankton in marine and freshwater ecosystem (Spector, 1984). The phytoplankton represent major constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem. The dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living as plankton or attached to sediments, sand, coral, or on the other aquatic plants (Hoek et al, 1995).

2.1 Taxonomy of Dinoflagellates

Dinoflagellates are classified as Protists under the division Dinophyta in botanical system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath & Saldarriaga, 2010). This is because some of the species are motile and heterotrophic while other species have cell walls and are photosynthetic (Cabrini et al, 2010). Approximately about 4500 species, 550 genera taxonomic data of dinoflagellates have been described and more than half of the species are in fossil form. From 2000 living species, more than 1700 are marine and about 220 are freshwater (Taylor et al., 2007). Dinoflagellates show a great range of forms but most of this diversity can be resolved to five basic types (Taylor, 1980) including the harmful species.

2.1.1 Dinoflagellates Morphology

Dinoflagellates are predominantly unicellular, eukaryotic, flagellated organisms that posses both photosynthetic and non photosynthetic characters. Their morphology is very

diverse. They can be divided into two types; depending on the point of insertion of the flagella namely desmokont and dinokont (Taylor et al, 2007). Desmokont is a dinoflagellate where the two flagella emerge at the cell apex. While dinokont, the flagella emerge from the middle of the cell (Bellinger & Sigee, 2010). The two flagella are inserted ventrally; one flagellum is longitudinal and housed in a sulcus. The transverse flagellum provides propulsion and the longitudinal flagellum provides direction (Tomas, 1997). In desmokonts, the pole are directed towards the swimming direction is anterior, the opposite pole is posterior, and the broad surfaces of the cell are lateral. In dinokonts, the side form which the flagella arise is ventral, the opposite surface is dorsal. Some examples of dinokont are Gymnodinium and Dinophysis. The other group, desmokont is Prorocentrum (Taylor, 1987). A dinoflagellate lacking cellulose plates is said to be athecate and cell covered only by a membrane (Cabrini et al, 2010). The plates may be thick, or so thin that they cannot be seen with the light microscope. There is generally an inverse relationship between the number and the thickness of the plates (Taylor, 1980). The number, shape and arrangement of these plates form a distinctive geometry known as plate tabulation, which is the main means for classification and they are a most important taxonomic criterion (Taylor, 1980).

Figure 1 : Basic Anatomy of Thecate, Dinokont Dinoflagellate. Modified After Evitt, 1985. (Macrae, 2007)

Dinoflagellate usually has a cellulose cell wall, perforated by many pores. Most forms have an equatorial groove that contains a ribbon flagellum. This groove separates the dinophyte's cellulose cell wall into two portions, the epicone and hypocone. The number, arrangement and thickness of the thecal plates can vary significantly. The cell surface ornamentation can be observed to identify the species such as pores, depressions, spines, ridges, and reticulations (Faust, 2002). Spines, wings, and horns may decorate the cell wall. The thecate forms have covering composed of three parts: epitheca (covering epicone), girdle plate (covering the girdle) and hypotheca (covering the hypocone) (Lebour, 1925). Dinoflagellates are mostly having a single nucleus, called the dinokaryon. The DNA is not organizing around nucleosomal histones, instead forming fibrillar chromosomes that are always condensed and divide through mitosis with an external spindle (Spector, 1984).

2.2 Distribution of Dinoflagellates

Dinoflagellates can be found with 90% in marine and 10% in freshwater in the aquatic environment (Taylor et al, 2007). The distribution of dinoflagellates depends on wind and currents, light, temperature, oxygen, pH, and nutrient uptake. Winds play an important role in the development of freshwater dinoflagellate blooms; whether in their vertical and horizontal distribution (Taylor, 1987). Dinoflagellates are commonly studied during their motile, planktonic stage; cyst-forming dinoflagellates which are known from all oceanic habitats. Temperature can influence photosynthetic rates and division rates, uptake and respiration rates and cell size of the dinoflagellates (Steidinger, 1997).

2.3 Ecology and Growth of Dinoflagellates

Approximately 1316% of living dinoflagellates produce a dormant resting cyst (Head, 1996). Cyst formation for dinoflagellates was basically related to the formation of seed to initiate red tides or Harmful Algal Blooms (HABs), existence approach due to environmental stress, resources for genetic recombination, direct source of toxicity and factor in bloom termination (Anderson et al., 2003). Therefore, dinoflagellates cysts provide information of mechanisms of spreading and reoccurrence of HAB (Sidharta et al., 2008). Dinoflagellates are the second most abundant form of autotrophic life in the marine ecosystem. As such, they are at the base of the food chain and provide food for herbivorous zooplankton and sessile benthic suspension feeders. They can be found either as freefloating planktonic dinoflagellate to benthic habitats as attached with the bottom from freshwater to estuaries and to hypersaline waters (Faust, 2002). Dinoflagellates are typically large-celled organisms such as Ceratium, Peridinium and Peridiniopsis. Dinoflagellate can vertically migrate, up during the day when the light is strongest, and down at night (Rapport, 1996). Dinoflagellates are the only photosynthetic organisms capable of bioluminescence (Taylor, 1987). Some species will act as parasite on other organisms and some species also support coral reef ecosystems through symbiotic associations. This type of dinoflagellate is called zooxanthellae.

2.4 Harmful Algal Blooms (HABs)

Mass productions of phytoplankton are known as algal blooms. Such blooms contain high concentrations of algal biomass. HABs causing discoloration of water are commonly

known as red tides (Hallegraeff, 1995). Dinoflagellate species such as Dinophysis, Alexandrium and Prodinium can contaminate shellfish with toxins, even at very low cell concentration (Hallegraeff, 2004). Harmful dinoflagellates are basically indistinguishable from other dinoflagellates from the same habitats in their responses to temperature, salinity, and light (Taylor, 1987). Freshwater dinoflagellate has poor salinity tolerance, where estuarine and some marine species can tolerate a wide salinity range (Taylor, 1987). There are different types of harmful dinoflagellate blooms. Most of the information is adapted from Camacho et al., (2006). a) Species which produce basically harmless irritation conditions (odours and/or discoloration of water) in sheltered bays that causes oxygen depletion. Example: Gonyaulax polygramma, Gymnodinium sanguinum, Noctiluca scintilans. b) Species that produce potent toxins that can find their way through the food chain to humans, causing a variety of gastrointestinal and neurological illnesses. Some of the species can contaminated the shellfish at very low cell concentrations. i. Diarrhetic Shellfish Poisoning (DSP); Mostly found in mussles, scallops, clams and gastropod. The main toxins were Okadaic acid, dinophysis toxins (DTXs), yessotoxins (YTXs) and pectenotoxins (PTXs). Example: Dinophysis acuta, D. acuminata, D. rotundata, Prorocentrum belizeanum, P. faustiae, P.lima. ii. Ciguatera Fish Poisoning (CFP) Ciguatera fish poisoning is the most common marine toxin disease worldwide. The primary toxin involve is ciguatoxin and mostly been contaminated if consume reef fish such as barracuda, grouper, and snapper. Example: Gambierdiscus toxicus, Ostreopsis

mascarenensis, Prorocentrum sp.

iii. Paralytic Shellfish Poisoning (PSP). It is associated with saxitoxin

(STXs); Example: Alexandrium acatenella, A. catenella, A. cohorticula, A. fundyense, A. fraterculua, A. minutum, A. tamarense, Gymnodinium catenatum, Pyrodinium bahamense var. compressum. iv. Neurotoxin Shellfish Poisoning (NSP); the toxins contain was brevetoxins (PbTxs). Detected to who consume oyster, clams, mussels, cockles, and whelks. Example: Karenina breve, and K. cf. breve from New Zealand.

c) Species which are nontoxic to humans but harmful to fish and marine invertebrates, especially in intensive aquaculture systems. The cells may cause damage or clog the gills of these animals. It is mostly attacked mussels and oyster. Example: Gymnodinium mikimotoi.

2.4.1 Impacts of HABs

HABs are natural phenomena, but their occurrence, geographic range and intensity appear to have increased since the 1970s and their economic impact is larger now compare to the past (Camacho et al., 2006). The increased economic impact of HABs is probably linked with the increased consumption of seafood and growth in coastal populations (Anderson et al., 2003). Symptoms occur generally as a consequence of consumption of contaminated seafood and direct human exposure to HABs.

3.0 MATERIALS AND METHODS

3.1 Study areas The study was carried out at Santubong River between December 2011 and April 2012. Santubong River is an important river that located in Santubong Village which is about 35 km from Kuching City. There is a lot of other village which depend on aquaculture industries and also transportation in Santubong area. Phytoplankton samples were collected from three stations along the Santubong River. The following months, the sampling was done with two other stations. The stations were; fish cages culture, shrimp aquaculture discharge and jetty of Fisheries Department. The location of each site was coordinated using Global Positioning System (GPS). All the locations were recorded in Table 1.

Table 1 : Sampling sites (station) along the Santubong River

Station 1 2 3

Location Jetty of Fisheries Department Fish Cage Culture Shrimp aquaculture discharge

Coordinate N 0142 85, E 11019270 N 0142 873, E1020126 N 0142 457, E 1020432

Figure 2: Maps of Santubong River

10

3.2 Phytoplankton Sampling and Water Quality Collection

Eighteen water samples were collected using Van Dorn water sampler with 20m mesh size plankton net at the surface of the water column. Three stations were selected; jetty of Fisheries Department, shrimp farm discharge and fish cage culture. On the first two months of sampling; December 2011 and January 2012, only one stations of sampling were done. The next sampling February to April 2012 were done in three stations with varies tidal range. The water samples are kept in the 1L Whirl-Pak and kept in the cooler box. Insitu water quality namely temperature and pH using pH meter, turbidity using turbidity meter, salinity was estimated using salinometer, and dissolved oxygen (DO) using DO meter. The transparency of water column was determined with a Secchi disk. The depth of the river is measured in every sampling using depth finder. The means of temperature, pH, salinity, DO, turbidity for each sampling station were obtain from three replicate readings. The water samples were transported to the laboratory and preserved with Lugols acidic solution. Some fresh samples were observed under the compound microscope immediately after field sampling. Lugols solution is added at a concentration of 1 mL per 100 mL sample (APHA, 1976). An advantage of Lugols solution is that flagellates preserved with it retain their flagella (Htzel & Croome, 1999). The water samples were let to settle for at least 24 hour. After that, the upper layer of sediment samples were siphoned off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration (Bangheri et al., 2010). The nutrients of the water samples are refer to the previous study done by Ling et al. (2010).

11

3.3 Phytoplankton identification and enumeration

One ml aliquot from each sample was poured in the counting slide with coverslips (24x20) and observed under compound microscope with 4X, and 10X objectives (Liliana, 2005). Phytoplankton were counted and identified to the lowest taxa under compound microscope Magnus Live model equip with connection to the laptop to capture the picture of phytoplankton found.

3.4 Statistical Data Analysis

Statistical analyses of data obtained from each station among months were determined using SPSS version 20 for Windows. Significant differences of these parameters at each sampling station monthly were tested with one-way ANOVA with P0.05 using Tukey statistical test (Liliana, 2005).

12

4.0 RESULTS

4.1 Qualitative Phytoplankton Composition

The phytoplankton checklist and contribution of different taxonomic groups to the total phytoplankton are presented in Table 2. A total of 89 taxa were identified in this study. From the list, 8 genera, consists of 17 species of dinoflagellates, 69 species of diatoms from 20 genera and blue-green algae. Diatoms were most occurred taxa during every months of sampling compare to dinoflagellates. Family Chaetoceraceae from centric diatom made up the higher number (2 genera, 13 species), followed by family from pennate diatom, Bacillariaceae (6 genera, 10 species).

70 60 50 Percentage % 40 Dinoflagellate 30 20 10 0 December January February March April Month Pennate Diatom Centric Diatom

Figure 3: Percentage Distribution of Phytoplankton

13

4.2 Dinoflagellates Assemblages

The species composition of dinoflagellates was lower than diatoms throughout the study period at all sampled locations. Some were recorded throughout the months and some were absent. Seventeen species of planktonic dinoflagellates belonging to 8 genera were recorded during this study. They occurred throughout the period of study with distinct monthly sampling. Ceratium is the common and distributed genus in the study areas with four species of were identified namely, Ceratium furca, Ceratium fusus, Ceratium trichoceros and Ceratium tripos. Meanwhile, the Dinophysis and Protoperidinium with three species each. The cell counts of C. furca were the highest during December 2011 and those Protoperidinium on April 2012 off Santubong River. Many species of potentially toxic dinoflagellates were found in this study. All of them occurred in low cell numbers. C. furca, C.fusus, Prorocentrum sigmoides, P. micans and Protoperidinium sp. can be found in every months of the sampling location. Individual of Pyrophacus, Pyrocystis lunula, and D.mile were less detected at all locations.

14

18 16 14 Number of Individual 12 10 8 6 4 2 0 December January February March April Month Figure 4: Number of Dinoflagellates Species identified in Monthly Sampling Ceratiaceae Dinophysiaceae Gymnodiniaceae Prorocentraceae Protoperidiniaceae Pyrocystaceae Pyrophacaceae

400 350 300 250 Individual per ml 200 150 100 50 0 Number of Individual

Family of Dinoflagellates

Figure 5: Number of dinoflagellates individual per ml along Santubong River.

15

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

Figure 6a: Representative Dinoflagellates (10X magnification). (a) Ceratium furca (b) Ceratium fusus (c) Ceratium trichoceros and Protoperidinium sp. (d) Dinophysis miles (e) Dinophysis caudata dividing (f) Dinophysis caudata (g,h) Protoperidinium sp.

16

(i)

(j)

(k)

(l)

(m)

(n)

(o)

(p)

Figure 6b: Representative Dinoflagellates (10X magnification). (i) Ceratium tripos (j) Gonyaulax sp (k) Gymnodinium sanguineum (l) Gymnodonium sp. (m) Prorcentrum micans (n) Prorcentrum sigmoides (o) Pyrocystis lunula (p) Pyrophacus sp.

17

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

Figure 7a: Representative Centric Diatoms. (10X magnification). (a) Chaetoceros sp. (b) Coscinodiscus sp. (c) Biddulphia Gray (d) Ditylum brightwelli (e) Hyalodiscus Ehrenberg (f) Rhizosolenia sp. (g) Skeletonema costatum (h) Cerataulina sp.

18

(i)

(j)

(k)

(l)

(h)

(m)

(n)

(o)

(p)

Figure 7b: Representative Pennate Diatom: (10X magnification). (i) Asterionella Hassal (j) Thallasiothrix Cleve & Grunow (k) Bacillaria paradoxa (l) Thalassionema sp. (m) Navicula sp. (n) Pleurosigma sp. (o) Pseudonitzchia sp. (p) Cylindrotheca closterium

19

Table 2: Taxonomic List and Occurrence of Dinoflagellates in Monthly Sampling

Dinoflagellates December Family Ceratiaceae Ceratium furca (Ehrenberg) Claparede & Lachmann, 1859 C. fusus (Ehernberg) Dujardin, 1981 C. trichoceros (Ehrenberg) Kofoid C. tripos (O.F. Muller) Nitzsch Family Dinophysiaceae Dinophysis sp. D. caudata Saville Kent,1881 D. miles var. Indica Bohm Family Gymnodiniaceae Gymnodinium sp. Akashiwo sanguineum Hirasaka 1922 Family Gonyaulacaceae Gonyaulax sp. Family Prorocentraceae Prorocentrum sigmoides Bhm P. micans Ehrenberg, 1833 Family Protoperidiniaceae Protoperidinium sp. P. pyriforme Peridinium Ehrenberg Family Pyrocystaceae Pyrocystis lunula Family Pyrophacaceae Pyrophacus sp. ++ + ++ + + ++ + + + + + +

Months of Sampling January February March ++ + + + + + + + ++ + + + + + + + + + + ++ + + + + + ++ +++ + + -

April +++ ++ +++ + ++ ++ + + + ++ + +++ ++ + +

Note: +: present ++: frequent +++: abundant

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Table 3: Other Phytoplankton Identified In The Samples along Santubong River.

Other Phytoplankton Centric Diatom Family Biddulphiaceae Biddulphia sp. B.Gray Family Chaetoceraceae Bacteriastrum hyalinum Lauder Chaetoceros capense C. constrictus C. curpiseris C. curvitus Cleve C. diversus Cleve C. decipiens Cleve C. Ehnrenberg C. lorenzianus Grunow C. peruvianus Brightwell C. pseudocurvisetus Mangin C. subtilis Cleve C. affinis Lauder Family Coscinodiscaceae Coscinodiscus sp. C. granii C. marginatus Family Thalassiosiraceae Cyclotella kutzing Skeletonema costatum Thalassiosira subtilis Family Lithodesmisceae Ditylum brightwelii D. Bailey Family Hemiaulaceae Eucampia zoodiscus Cerataulina sp. Family Rhizosoleniaceae Guinerdia sp. Rhizosolenia sp. R. segitera Proboscia alata Family Hyalodiscidae Hyalodiscus Ehrenberg December

Months of Sampling January February March April

++ ++ + + + + ++ + + + + + + + + ++ + + +++ + ++ + + + + 21

++ ++ + + + + ++ + + + + + + + + + + + + ++ + ++ + + + + + +

++ + + + + + + + ++ + ++ ++ + ++ + + + + + ++

+ + + + ++ ++ + + + ++ +++ ++ ++ + + ++ + + + + ++ + ++ +

+ ++ + + + + +++ + + + + + + + + + + + + +++ + ++ + ++ + ++ ++ + ++ +

Family Leptocylindraceae Leptocylindrus danicus Cleve Family Eupodiscaceae Odontella sp. O. mobiliensis Triceratium sp. Family Melosiraceae Paralia Heiberg Stephanopyxis Melosira cf. Spaerica M. granulata M. sulcata Family Streptotheca Streptotheca thamesis Family Corethraceae Corethron sp. Pennate Diatom Family Fragilariaceae Asterionella Hassal Diatoma. Fragilliariopsis Synedra Family Naviculaceae Amphiprora Diploneis Gyrosigma G. auminatum Navicula N. didyma N. miniscul Family Thalassionemataceae Thalassionema T.frauenfeldii T.nitzschioides Thalassiothrix Cleve n Grunow Family 18 Bacillariaceae Bacillaria paxillifer Cylindrotheca Closterium Nitzchia. N. sigma N. seriata Pleurosigma P. W. Smith

+ ++ ++ + + + ++ -

+ ++ ++ + + + + -

+ ++ ++ ++ ++ + + + -

+ ++ ++ + ++ + + + + -

++ ++ ++ + + + + + + + +

+ + + + + ++ ++ ++ + + + + +
22

++ + + + + + ++ + ++ +++ + + + + +

+ + + + + + + ++ + + ++ + ++ ++ + + + + + +

+ + + + + + + + + + + ++ + ++ +++ + + + + + +

+ + + + + + + + + ++ + ++ + +++ +++ + + ++ + + ++ +

Pseudo-nitzchia P. austrialis Family 19 Pinnulariaceae Pinnularia alpine Blue-Green Algae Family Dictyochaceae Dictuocha Ehrenberg Division Chlorophyta Scenedesmus spp. Oscillatoria princeps Spicule Anabaena Octatis Ehrenberg

+ +

+ + +

+++ + +

+ + -

++ + + -

++ + + + -

++ + + + +

+ + + +

Note: +: present ++: frequent +++: abundant

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4.2.1 Dinoflagellates Taxonomic Descriptions

Dinophyceae are unicellular eukaryotic organisms which some species that are able to move with its flagella. Dinoflagellates are categorised into two types: armoured and unarmoured (Taylor, 1987). The species can be observed in a dorsal side where the cingulum is visible. In a ventral side, where perpendicular to the cingulum, the longitudinal sulcus lies towards the antapex (Cabrini et al, 2010).

Family: CERATIACEAE Ceratium has been suggested as an excellent, if not the best, dinoflagellate genus to use for biogeographic study and as a tool for defining ocean currents and temperature ranges and may be valuable in studies of global change (Dodge & Marshall, 1994).

Ceratium furca (Ehrenberg) Claparde & Lachman The species is known to be found in warm to cold temperate waters. It is a large species with two unequal hypothecal horns which are serrated (Steidinger, 1997). It is covered by an armour-like cell wall or theca and composed of many plates. It also has one horn that shorter than the other. They are non-toxic and are necessary for the food web. However, they can cause a red tide if conditions allow for excessive blooming (Hardy, 2010).

Ceratium fusus (Ehrenberg) Dujardin It is a long thin shaped Ceratium and armored species of dinoflagellate. From a swollen body the apical horn is long, slightly tapers and has a slight curve (Steidinger, 1997). The long horn keeps it floating and prevents it to move quickly (Hardy, 2010). It is red-tide

24

species, but non-toxic. It can cause oxygen depletion when proliferate in high concentration.

Ceratium trichoceros (Ehrenberg) Kofoid Apical and hypothecal horns curve widely in parallel plane (Steidinger, 1997). Have small body. It can be found distributed in coastal and oceanic. It is also cosmopolitan in warm temperate to tropical waters.

Ceratium tripos (O.F. Mller) Nitzsch Large cell triangular epitheca that abruptly forms straight apical horn. The hypothcea is smooth and curved. The hypothcel horns are short; almost level with the cell body. Ceratium tripos is sometimes parasitized by other protists.

Family: DINOPHYSIACEAE

Dinophysis caudata Saville-Kent Dinophysis caudata is an armoured, marine, planktonic dinoflagellate species (Bennion et al, 2005). It is a bloom-forming species associated with massive fish kills (Faust & Gulledge, nd). D. caudata is a photosynthetic species with chloroplasts and a large posterior nucleus. Cell is medium sized and has a characteristic posterior finger-like process. Dorsal side is slightly concave in the anterior half, whereas the ventral margin in lateral profile is generally straight along the main body (Steidinger, 1997). The cells usually occur in pairs and attached at dorsally. This species is widely distributed throughout the world from temperate to tropical water.

25

Dinophysis miles var. indica Bhm The cells had two posterior projections that extended from the end of the hypotheca (Qiu et al., 2011). It is known as a toxic marine species. It is found accumulates in high level of diarrhetic shellfish poisoning toxins in the green mussel Perna viridis during a bloom at Saipan Bay, Panay Island, Philippines (Anderson et al., 2003). Toxic strains only reported from the Philippines (Marasigan et al., 2001). The species were widely distributed in tropical seas (Moestrup, 2010).

Family: GONYAULACACEAE Gonyaulax sp. Gonyaulax sp. is an armoured type of dinoflagellates. The body is covered within thecal plates. It has two distinct grooves, where the flagella are situated. Some of the species is known toxic species (Hardy, 2010).

Family: GYMNODINIACEAE Gymnodinium sp. Gymnodonium sp. is an unarmored species which covered with a thin pellicle, with both types of flagella located in sulcus and girdle (Lebour, 1925). It is single cell formed. The cingulum was usually equatorial with descending displacement (Steidinger, 1997). It can be found from oceanic to estuarine; warm temperate to tropical.

26

Akashiwo sanguinea K.Hirasaka, 1922

It is planktonic marine species of dinoflagellates. The cell is unarmoured which the size of the hypotheca and epitheca almost equal (Steidinger & Tangen, 1997). It has large, nucleus in the centre and numerous chloroplasts. It can roduce potent toxins that if consumed by humans, cause gastrointestinal and neurological illnesses (Camacho et al., 2006). Distribution: Cosmopolitan in temperate to tropical coastal and estuarine waters.

Family: PERIDINIACEA

Peridinium Ehrenberg

This genus is principally freshwater or brackish. Peridinium has thick, armored plates that are often lobed and ornamented. The hemisphere with the longitudinal groove ends with two projections. The other hemisphere ends with a single projection.

Family: PROROCENTRACEAE

Prorocentrum sigmoides Bhm

Elongated cell, sometimes in "S" shape. Has apical spine sharp and tiny; protruded from the anterior side. It is non-toxic but will harmful to marine organisms due to oxygen depletion at high cell concentration.

27

Prorocentrum micans Ehrenberg

It is an armoured, planktonic, photosynthetic species and also can cause bloom in mostly coastal water. Although P. micans is capable of forming extensive blooms, it is usually considered harmless (Taylor, 1987). The cell is in tear-drop shape, with broad in the middle. The adjacent to the flagellar pore is slightly small. There is no cingulum or sulcus and having well developed apical spine. This species are commonly found in neritic and estuarine waters, but it s also found in oceanic water (Dodge & Marshall, 1994). Thats mean; this species can tolerate various range of salinity. P. micans is one of the most common and varied species in the genus Prorocentrum (Faust & Gulledge, nd).

Family: PROTOPERIDINIACEAE

Protoperidinium sp.

Protoperidinium sp is an armored species (Cabrini et al, 2010). It is a round to polygon shaped cell with one apical horn and two antapical horns. The spines can be seen and it has two obvious grooves: the sulcus (vertical) and the cingulum (horizontal). The surface is reticulated.

Peridinium pyriforme Paulsen

The Peridinium cell divided by a cingulum or transversal sulcus that can be circular or elicoidal. The cell is pear in shape. The hypotheca is rounded bearing two winged spines. The sulcus is broadens and slightly posteriorly. It is mostly found in marine ecosystem.

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Family: PYROCYSTACEAE

Pyrocystis lunula J.Schtt

It is marine and bioluminescent type of dinoflagellates. It is unarmored, and has crescent moon-shaped cells. Pyrophacus cells are biconvex, lens-shaped and compressed anteroposteriorly (Balkis & Koray, 2001). Chloroplasts can be seen present in the cell. Distribution: Oceanic and coastal. Warm temperate to tropical waters. Family: PYROPHACACEAE

Pyrophacus sp.

An armored species and has large biconical, lens-shaped shapes (Steidinger, 1997). The thecal plates are thin, flat and compressed anteroposteriorly (Balkis & Koray, 2001). The species was well distributed from neritic to estuarine water.

29

4.3 Diatoms Assemblages

60.9 % of centric diatoms and 25.32% of pennate diatom identified in this research study. The most common taxa in centric diatoms were from genus Coscinodiscus and Melosira with thirteen and four species, respectively. Genus Thalassiothrix and Thalassionema were the most common pennate diatoms composed of four and eight species in those genera. The composition of centric diatom was higher than composition of pennate diatom in all the samples.

70 60 Percentage % 50 40 30 20 10 0 April March February January December

Figure 8: Percentage Abundance of Diatoms along Santubong River

30

4.3.1 Diatom Taxonomic Description

Diatoms are very distinct group of algae, identifiable under light microscope by their own yellow brown colouration and the presence of a thick silica cell wall (Bellinger & Sigee, 2010). Diatom occurs as single cells or chains or other loose aggregates. They are extremely abundant both in in marine and freshwater ecosystems thus, they play major role in primary productivity for aquatic organisms (Wetmore, n.d). They can be planktonic or benthic, as presented in variable ways liable on latitude, and occur in all the periods (Cabrini et al., 2010). The diatoms were classified into two types; centric and pennate diatoms and are mostly unicellular, with some can formed chain of cells and colonial aggregations (Fareha et al., 2010). They can be presented in two orders; Biddulphiales (centric) and Bacillariales (pennate) (Cabrini et al., 2010).

31

4.4 Physico-chemical parameters 4.4.1 pH The pH values in the Santubong River were relatively alkaline. The range of pH value is between 7.3 and 8.0. The highest mean of pH value was recorded in Station 1, 8.130.01 in month of April while the lowest is at Station 1 on the February with a pH value of 7.30.03. Statistical analysis ANOVA indicates that the pH were not significantly difference between months (p0.05).

Figure 9: The Mean pH Value of Three Sampling Stations From December 2011 To April 2012.

32

4.4.2 Temperature

Temporal variations of temperature were relatively warm in every months of sampling stations (29.1-32.2C). The temperature is the highest on April, 32.21.3C at Station 2 and lowest was on February, Station 1 with 29.01.24 C. The temperature of Station 1 is slightly decreasing from December, 30.4C and February, 29.4 C. In February, the sampling is conducted during the low tide, and the temperature recorded was slightly increases until April. Statistical analysis ANOVA shows that, there were significantly differences of temperature (P0.05) between January, February and March of sampling with April.

Figure 10: The Mean Value of Temperature (C) In Three Sampling Stations from December 2011 to April 2012

33

4.4.3 Salinity

Salinity varied from 27 to 31 PSU, monthly with slightly decreasing from March to April in each station. The salinity is the highest on December 2011 with 31.30.15 PSU and the lowest also in Station 1 on February, about 261 PSU. Statistical analysis indicates there is no significant difference (P0.05) of salinity between months (Table 5).

Figure 11: The Mean Value of Salinity in Three Sampling Locations from December 2011 to April 2012.

34

4.4.4 Dissolved Oxygen

Dissolved oxygen concentrations altered significantly throughout the sampling period (2.1-8.2 mgL-1) where the highest concentration is at Station 1 on April with 8.20 0.01 mgL-1 and the minimum is at Station 2 on the February with only 2.1 0.38 mgL-1. There is significantly difference of dissolved oxygen between months shows in statistical analysis. The sampling on February has significantly difference (P0.05) with other month of sampling.

Figure 12: The Mean Value of Dissolved Oxygen (DO) in Three Sampling Stations from December 2011 to April 2012.

35

4.4.5 Turbidity

The turbidity shows a fluctuation between months and the stations. There is very high in turbidity in the month of February (64 13 NTU). The water is very turbid due to the low tide. The lowest (5.9 2.5 NTU) turbid value occurs on the January at Station 1. Statistical analysis ANOVA indicates that there was significantly difference (P0.05) of turbidity in every month of sampling.

Figure 13: The Mean Value of Turbidity in Three Sampling Stations from December 2011 to April 2012.

36

5.0 DISCUSSION

Aquaculture is the fastest growing sector in the food industry (Kutty, 2005). In Malaysia, fish cage farm and pond culture activities are mostly carried out in coastal aquatic environments, especially river mouths, estuaries, mangroves or any other types of brackish water bodies (Lokman et al., 1996). Santubong River, where the study site conducted, is in the Kuching City. It was one of the important rivers that provide opportunity for aquaculture activities and the establishment of many fish cage farm. Anthropogenic activities such as construction, sand dredging including other traditional activities compiled with aquaculture discharge contributes to eutrophication process in the area. There is high demand of oxygen level, high concentration of nutrient and low oxygen dissolved reported by Ling et al. (2010). There was also high number of chlorophyll-a (Chl-a) concentration in that area especially in shrimp farm discharged (Ling et al., 2010). Human activities such as sewage discharge and run-off from agriculture and aquaculture are common sources of these nutrients (Anderson et al., 2002). Three stations were selected in this study; (i) (ii) (iii) Fisheries Department jetty, Shrimp farm discharge and Fish cage farm.

Shrimp farm that were constructed at the estuaries that contributes the major effluent to the river. For fish culture, the main impacts were the increasing nitrate, phosphorus, and organic matter that enrich water and underlying sediment. Wastewater discharge at these stations increased nutrient loading which stimulated the growth of algae and bacteria (Ling et al., 2010), thus, increased the amount of Chl-a concentrations.
37

Phytoplankton compositions in all stations were consisted majorly by diatoms, followed by dinoflagellates and other phytoplankton. The growth of phytoplankton is influenced by several factors: light, temperature, nutrients and grazing (Burford, 1997). Eight genera of dinoflagellates were recorded in this study. They were Ceratium, Dinophysis, Prorocentrum; P. micans, Protoperidinium, Gymnodinium, Gonyaulax, Pyrocystis and Pyrophacus. The occurrence of potentially toxic species such as Dinophysis ;D.caudata, Prorocentrum; P. micans, Protoperidinium and Gymnodinium were lower number of individual during the study. The other common species encountered during the study period was Prorocentrum micans and it was found in every month of sampling. The genus Ceratium has shown more diverse in species composition (Table 2). The four common species were Ceratium furca, C. fusus, C. trichoceros and C. tripos. The genus Ceratium were frequently dominated the coastal phytoplankton community and contributes significantly to annual primary production in coastal water (Dodge & Marshall, 1994). Ceratium represents the important food sources for zooplankton compared to other dinoflagellates (Nielson, 1991). Ceratium furca is the common species found in every sampling month and showed the highest number of species was on the April 2012. Ceratium does not produce toxin but recently, extensive blooms of the species were reported along the Pacific coast of central Japan in 1997 and cause severe damage to local fisheries (Machida et al., 1999). The most famous was a bloom of Ceratium tripos in New York Bright in 1976, which was related with prolonged bottom-water anoxia and fish/shellfish kills (Mahoney & Steimle, 1979). In December; the salinity showed the highest value with 31.1 PSU. Ceratium furca is the highly-euryhaline species (Alkwari & Ramaiaah, 2010). It can tolerate high salinity water and wide range of temperature (Baek et al., 2005). In addition, Dinophysis caudata was also identified in dividing form in the samples. Dinophysis caudata is a very
38

distinctive species and reproduce by binary fission. There was red tides associated with mass mortality of fish have been reported in the Gulf of Thailand and Seto Inland Sea in Japan related to this species (Okaichi, 1967). But in this sampling location, the numbers of species were small and might not causing red-tide in the area. The depth of water was about 6.5 meters with the secchi disk transparency recorded about 6.0 meters. The transparency increase the amount of sunlight penetrates into the river allows the growth of phytoplankton. Most of the dinoflagellates were photosynthetic and manufactured their own food using the energy from sunlight, and providing a food source for other organisms (Waggoner & Speer, 2006). Skeletonema costatum and Coscinodiscus sp. were very abundant in this month. Skeletonema were usually found in neritic water and were identified as a harmful species. It is most abundant in brackish water (National Science Council, 2004). The number of species was rather low to be harmful for other aquatic life. Diatoms are one of the most important groups of producer organisms in the marine ecosystem (Bennion et al., 2005). It contributes major composition of phytoplankton besides dinoflagellates as shown in Figure 3. Diatoms have the larger number of genera in all sampling station and months. Centric diatoms seem more identified in the study period than pennate diatoms. The most appeared diatoms were Chaetoceros. Chaetoceros is mainly characterized by its chain-forming cells and their setae structures. (Sang & Jin, 2011). Six species of Chaetoceros were identified in this study (Table 3). On January 2012, the sampling was conducted in the morning with mean temperature was about 29.2C. Dissolved oxygen was the highest (8.53 mg/l). This might because of the high tide. The number of Ceratium species quite low in the February as compared to January and December, while the number of Dinophysis was higher on the

39

both month (Table 2). Dinophysis plays dual roles in the marine ecosystems: as primary (photosynthetic) and secondary (heterotrophic) producers (Qiu et al., 2011). Fortunately, Dinophysis miles were identified only on January and were not found in the other sampling months. The species were known as a toxic species which contain okadaic acid and characterized by elongated dorso-posterior region equal to or longer than the posterior process (Sadaff & S.M., 2010). During low tide, the number of phytoplankton was the lowest. This was shown in Table 2, on the month of February. Most of dinoflagellates cannot be identified in the sampling stations because the water was very turbid with small amount of dissolved oxygen concentration. High turbidity cause low penetration of microalgae that influence the growth of phytoplankton. During low tide or dry season, the flow of water decrease and the nutrient supplies were limited (Anderson et al., 2002). Moreover, the weather on that day was raining, thus it will lead to run-off from the land to the river. This will expose the soil to erode and increased the sedimentation. Rains also have significant lowered the salinity of the river and dinoflagellates species have tolerate changes of salinities (Liliana, 2005). As shown in Figure 11, dissolved oxygen (DO) drop promptly (2.1-2.8 mgL-1), which was the lowest value recorded. At this DO level, it causes adverse affect to the functioning and biological communities (Chapman, nd). There were significant different of turbidity in all month of sampling. High turbidity reduces the salinity, decrease the temperature, DO and pH (Perumal et al., 2008). Pyrocystis were rare species in this study period. This species can only be identified in this month of sampling. Unlike other phytoplankton, diatom from the genus Odontella was frequently identified in this month of sampling. On March sampling, there were two more stations included; fish cage farm and shrimp farm discharge. Statistical analysis shows that all the physico-chemical parameters
40

are significantly difference between months (Table 4). By station comparison, there are no significant differences (P0.05). This might be due to the sampling location was not far from the others. The dominant species on this month was Protoperidinium sp. (Figure 4). Pyrophacus was also identified during this period. The sampling was done during high tide. The depth of the water was the highest on the station 2 with 13.6 meter. High tide cause water from the ocean to move in and out and it will dilute and carry away contaminants such as the ammonia excreted from the fish culture. In the month of April, the amount of phytoplankton is the highest. The temperature and the pH value were the highest in this month. Yoo (1991) stated that pH is the leading factor correlating with the abundance of dinoflagellates in Masan Bay, Korea. He also suggests pH is an important factor contributing to the regulation of dinoflagellate blooms in a coastal marine environment. Most of freshwater dinoflagellates preferred alkaline water bodies and only few of them prefer acidic water bodies (Pollingher, 1978). Several dinoflagellates can tolerate in the high pH value, for example Gymnodinium (Figure 4). The number of Gymnodinium was increasing from first sampling (December 2011) but drop in the last sampling (April 2012) might be due to the increasing value of pH. The salinity decreasing from March to April because of the river is high tide and lot of water inputs from river both months. In this study, the possible main sources of nutrients were: fertilizers, shrimp food (nitrogen and phosphorus) and shrimp excreta (ammonia) (Burford, 1997). According to Ling et al., the concentration of phosphorus in the river was in the range of 0.16 to 0.2mgL-1. The urine and feaces from the aquatic animals can cause high content of ammonia. Ammonia is the main nitrogenous waste that is produced by fish via metabolism and is excreted across the gills (Cao et al., 2007). The effect of organic loading influence that changes in phytoplankton biomass, composition, structure and dominance.
41

Outbreaks of shellfish poisoning cases due to microalgal blooms have caused serious economic losses in the fisheries industries in many countries. Public health worries arise with increasing poisoning cases due to the consumption of contaminated bivalve. Although Malaysian waters, especially Sarawak have no record bout this phenomenon, the awareness and the proper management ad mitigation should be measured on the water quality of Sarawak River to avoid disease and maintain healthy aquaculture industry. As mitigation to safeguard the aquaculture industries in the country, base line data on the occurrence of toxic microalgae should be documented.

42

6.0 CONCLUSION

Santubong River shows a variety of phytoplankton present in water bodies especially for diatoms. Eight dinoflagellate genera namely Ceratium, Dinophysis, Gymnodinium, Gonyaulax, Prorcentrum, Protoperidinium, Pyrocystis and Pyrophacus were recorded. Another 72 genera were belongs to diatoms. Ceratium furca and a diatom Chaetoceros sp. are the most common species found. A few potentially toxic dinoflagellates species have been detected. They were Dinophysis, Prorocentrum and Gymnodinium. While, dinoflagellates that were identified showed a diverse community. The genus Ceratium shows high species composition in all sampling location. All the taxonomic data showed that the presence of potential toxin producers and bloom-forming species of dinoflagellates is in low concentration. The water physico-chemical parameters range from 29.1-32.2C for temperature, 26-31.3 PSU for salinity, 7.3-8.0 for pH, 2.18.2mgL-1 for dissolved oxygen (DO) and 5.9-64 NTU for turbidity of Santubong River. Without proper management, the nutrient loading can cause eutrophication occurrence and it will lead to red tide phenomenon. This phenomenon might affect the aquatic ecosystem in the river and the water quality in the area will be worsen and cause negative impact to aquaculture industry in Santubong River.

LIMITATION AND RECOMMENDATION The observations were taken only during six months, therefore, the correlation of water parameters and number of species could not be described. During the species identification process, some of the species were hardly identified. It consume lots of time. However, species like Dinophysis and Ceratium were easily identified due to their morphological characteristics. There were also some problems occurred on the first
43

sampling where we cannot found the boats available at the sampling area. Furthermore, the boat rental was very costly and tooks time to get approval of financial from the faculty. It tooks time to find the boats and causing some data were not available in this research study which was conducted in only a few months.

44

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Sang, D.L. & Jin, H.L. (2011). Morphology and taxonomy of the planktonic diatom Chaetoceros species (Bacillariophyceae) with special intercalary setae in Korean coastal waters. The Korean Society of Phycology. Algae 2011, 26(2): 153-165 Siddartha, B.R., Panggabean, M.G.L., & Mizushima, K. (2008). Horizontal Distribution of Dinoflagellates Cysts in Surface Sediments of Jakarta Bay: Some Preliminary Result. Marine Research Indonesia. Vol 33, No.1. Spector, D.L. (1984). Dinoflagellates. Academic Press, Orlando Steidinger, K.A. (1997). Dinoflagellates. In: Tomas, C.R. (eds.) 1996, Identifying Marine Phytoplankton. Academic Press Inc. p.387-584 Taylor, F.J.R. (1980). Review on Dino Evolution. Biosystem. pp 65-108. Taylor, F.J.R. (ed.) (1987). The biology of Dinoflagellates. Vol 21. Blackwell Scientific Publication, Boston. pp 399-529. Taylor, F.J.R., Hoppenrath M., Saldarriaga J.F (2007). Dinoflagellate Diversity and Distribution, Biodiversity Conservation, Vancouver, Canada. Tomas, C.R. (1997). Dinoflagellates: Identifying Marine Phytoplankton. Academic Press. USA. Waggoner,B. & Speer, B.R., (2006). Introduction to Dinoflagellata. University of California. Retrived on October 11, 2011, at http://www.ucmp.berkeley.edu/protista/dinoflagellata.html. Yoo, K. I. (1991). Population Dynamics of Dinoflagellate Community in Masan Bay with a Note on the Impact of Environmental Parameters. Marine Pollution Bulletin. Korea.

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APPENDIX

Table 4: ANOVA Table for Physico-chemical Parameters

ANOVA Sum of df Squares pH Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total 1.252 .512 1.764 18.674 15.024 33.699 65.469 26.453 91.922 5698.003 1806.128 7504.130 90.689 9.079 99.768 4 28 32 4 28 32 4 28 32 4 28 32 4 28 32

Mean Square .313 .018 4.669 .537 16.367 .945 1424.501 64.505 22.672 .324

F 17.119

Sig. .000

8.701

.000

Temperature

17.324

.000

Salinity

22.084

.000

Turbidity

Dissolved Oxygen

69.926

.000

49

Table 5: Multiple Comparisons for Physico-Chemical Parameters

Tukey HSD Dependent Variable

(I) Month

(J) Month

Mean Difference (I-J) .1733 .3233* -.0178 -.1822 -.1733 .1500 -.1911 -.3556* -.3233* -.1500 -.3411* -.5056* .0178 .1911 .3411* -.1644 .1822 .3556* .5056* .1644 .9000 .8667 .5444 -.9556 -.9000 -.0333 -.3556 -1.8556* -.8667 .0333 -.3222 -1.8222* -.5444 .3556 .3222 -1.5000* .9556 1.8556* 1.8222* 1.5000* .2667
50

Std. Error

Sig.

December

January

pH

February

March

April

December

January

Temperature February

March

April Salinity December

January February March April December February March April December January March April December January February April December January February March January February March April December February March April December January March April December January February April December January February March January

.1104 .0902 .0902 .0902 .1104 .0902 .0902 .0902 .0902 .0902 .0637 .0637 .0902 .0902 .0637 .0637 .0902 .0902 .0637 .0637 .5981 .4883 .4883 .4883 .5981 .4883 .4883 .4883 .4883 .4883 .3453 .3453 .4883 .4883 .3453 .3453 .4883 .4883 .3453 .3453 .7936

.528 .010 1.000 .282 .528 .471 .240 .004 .010 .471 .000 .000 1.000 .240 .000 .102 .282 .004 .000 .102 .568 .408 .797 .313 .568 1.000 .948 .006 .408 1.000 .882 .000 .797 .948 .882 .001 .313 .006 .000 .001 .997

95% Confidence Interval Lower Upper Bound Bound -.148 .495 .061 .586 -.280 .245 -.445 .080 -.495 .148 -.113 .413 -.454 .072 -.618 -.093 -.586 -.061 -.413 .113 -.527 -.155 -.691 -.320 -.245 .280 -.072 .454 .155 .527 -.350 .021 -.080 .445 .093 .618 .320 .691 -.021 .350 -.843 2.643 -.556 2.289 -.878 1.967 -2.378 .467 -2.643 .843 -1.456 1.389 -1.778 1.067 -3.278 -.433 -2.289 .556 -1.389 1.456 -1.328 .684 -2.828 -.816 -1.967 .878 -1.067 1.778 -.684 1.328 -2.506 -.494 -.467 2.378 .433 3.278 .816 2.828 .494 2.506 -2.046 2.579

January

February

March

April

December

January

Turbidity

February

March

April

February March April December February March April December January March April December January February April December January February March January February March April December February March April December January March April December January February April December January February March January February March April December February March April December

3.6889* 1.5111 3.5778* -.2667 3.4222* 1.2444 3.3111* -3.6889* -3.4222* -2.1778* -.1111 -1.5111 -1.2444 2.1778* 2.0667* -3.5778* -3.3111* .1111 -2.0667* 5.7833 -28.0278* .5633 .9533 -5.7833 -33.8111* -5.2200 -4.8300 28.0278* 33.8111* 28.5911* 28.9811* -.5633 5.2200 -28.5911* .3900 -.9533 4.8300 -28.9811* -.3900 .5600 3.7211* .2322 -.3011 -.5600 3.1611* -.3278 -.8611 -3.7211*
51

.6480 .6480 .6480 .7936 .6480 .6480 .6480 .6480 .6480 .4582 .4582 .6480 .6480 .4582 .4582 .6480 .6480 .4582 .4582 6.5577 5.3543 5.3543 5.3543 6.5577 5.3543 5.3543 5.3543 5.3543 5.3543 3.7861 3.7861 5.3543 5.3543 3.7861 3.7861 5.3543 5.3543 3.7861 3.7861 .4649 .3796 .3796 .3796 .4649 .3796 .3796 .3796 .3796

.000 .165 .000 .997 .000 .330 .000 .000 .000 .000 .999 .165 .330 .000 .001 .000 .000 .999 .001 .901 .000 1.000 1.000 .901 .000 .864 .894 .000 .000 .000 .000 1.000 .864 .000 1.000 1.000 .894 .000 1.000 .749 .000 .972 .930 .749 .000 .908 .185 .000

1.801 -.377 1.690 -2.579 1.534 -.643 1.423 -5.577 -5.310 -3.513 -1.446 -3.399 -3.132 .843 .732 -5.466 -5.199 -1.224 -3.402 -13.322 -43.628 -15.036 -14.646 -24.889 -49.411 -20.820 -20.430 12.428 18.211 17.560 17.950 -16.163 -10.380 -39.622 -10.641 -16.553 -10.770 -40.012 -11.421 -.795 2.615 -.874 -1.407 -1.915 2.055 -1.434 -1.967 -4.827

5.577 3.399 5.466 2.046 5.310 3.132 5.199 -1.801 -1.534 -.843 1.224 .377 .643 3.513 3.402 -1.690 -1.423 1.446 -.732 24.889 -12.428 16.163 16.553 13.322 -18.211 10.380 10.770 43.628 49.411 39.622 40.012 15.036 20.820 -17.560 11.421 14.646 20.430 -17.950 10.641 1.915 4.827 1.338 .805 .795 4.267 .778 .245 -2.615

December Dissolved Oxygen January February

January -3.1611* March -3.4889* April -4.0222* December -.2322 January .3278 March February 3.4889* April -.5333 December .3011 January .8611 April February 4.0222* March .5333 *. The mean difference is significant at the 0.05 level.

.3796 .2684 .2684 .3796 .3796 .2684 .2684 .3796 .3796 .2684 .2684

.000 .000 .000 .972 .908 .000 .298 .930 .185 .000 .298

-4.267 -4.271 -4.804 -1.338 -.778 2.707 -1.315 -.805 -.245 3.240 -.249

-2.055 -2.707 -3.240 .874 1.434 4.271 .249 1.407 1.967 4.804 1.315

Table 6: The Depth of Water Column and its Transparency

Months December January February Station 1 Station 2 Station 3 March Station 1 Station 2 Station 3 April Station 1 Station 2 Station 3

Water depth (m) 6.5 5.8 5.4 11 5.2 9.7 13.6 5.8 7.05 12.9 2.7

Secchi depth (m) 6.23 5.1 3.5 4.56 4.5 7.3 5.05 4.44 6.05 9 0.53

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