รายงานประจำปี 2552 สถาบันวิทยาศาสตร์ทางทะเล

3Department of Biology, Faculty of Science, Faculty of Medicine, Chiang Mai University, Mahidol Unversity, Bangkok 10400 Thailand Chiang Mai 50200, Thailand. Abstract 3Institute of Marine Science, Burapha University, Little is known of reproductive patterns Bangsaen, Chon Buri, Thailand. of molluscs inhabiting mangroves. Yet this is one of the dominant environments in tropical intertidal Abstract areas and molluscs are one of the dominant Padina minor Yamada, a brown alga faunal groves in mangroves. Several species of which is found to be abundant on both sides of the gastropod genus Littoraria are adapted spe- the coastal area of the gulf of Thailand and the cifically to life in mangrove environments. The Andaman Sea was examined for gastroprotective present paper examines reproductive periodicity activity. The aqueous extract of P. minor in four species of Littoraria from Ang Sila, Thai- (Aq. P) given orally to rats at the doses land, from June 2003 to June 2004. Monthly of 100, 200 and 500 mg/kg significantly inhibited samples were made to measure reproduction gastric ulcer formation induced by (1) restraint and growth. Ripe or partly spawned occurred water immersion stress (2) acid ethanol (HCl / throughout study, suggesting that these species EtOH), (3) indomethacin and (4) histamine. In the are capable of either partially spawning frequently pylorus-ligated rat experiment, the Aq. P caused or rapidly returning to reproductive condition af- a decrease of the total acidity and an increase ter spawning. The major spawning occurred in in gastric pH. Additionally, the Aq. P could not August-September 2003 and January to Febru- preserve the mucus content in the gastric wall ary 2004. Littoraria palescens had a clear growth of rats with gastric ulceration induced by HCl / pattern with a one-year life cycle. In contrast, EtOH. Results were obtained from the isolated there was apparent recruitment failure in L. mel- guinea-pig right atrium experiment of which the anostoma and the combination of L. articulata inhibitory effects of cimetidine (Histamine H2- and L. strigata. antagonist) and the Aq. P on histamine-induced chronotropic responses were determined and Gastroprotective Activity of Padina these results suggest that they share a similar minor Yamada mechanism. The findings therefore indicate that P. minor possesses a gastroprotective activ- Doungporn Amornlerdpison1 , ity, which involves an anti-secretory mechanism Yuwadee Peerapornpisal1, Tawat Taesotikul2 , mediated via histamine H2-antagonism. The Thidarat Noiraksar3, and Duangta Kanjanapothi2 polysaccharide present in the P. minor is likely to contribute to the gastroprotective activity. 1Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand. 2Department of Pharmacology,

Apocyclops ramkhamhaengi sp. Biological properties of lectins in nov. (Copepoda: Cyclopoida) in marine sponges from a culture originating from brackish Chonburi Province, Thailand waters of Chang Island, Trat Province, Thailand Janjarus Watanachote1, Maywarin Chaichareon2 and Suriyan Tunkijjanukij3,4 Supawadee Chullasorn1, Pawana Kangtia1, Khwanruan Pinkaew2, and Frank D. Ferrari3 1Institute of Marine Science, Burapha 1Department of Biology, Faculty of Science, University, Chonburi, 20131, Thailand 2Department of Biology, Faculty of Science, Ramkhamhaeng University, Bangkapi, Burapha University, Chonburi, 20131, Thailand.D Bangkok 10240, Thailand Department of Marine Science, Faculty of 2Institute of Marine Science, Burapha University Fisheries, Kasetsart University, , Bangsaen, Chonburi Province 20131, Thailand Bangkok 10900, Thailand. 3IZ/MSC; MRC-534, National Museum of 4Center for Agricultural Biotechnology, Natural History, Smithsonian Institution, 4210 Kasetsart University, Kamphaeng Saen Campus, Nakorn Pathom 73140, Thailand. Silver Hill Rd., Suitland, MD 20746, USA Abstract Abstract In some invertebrate marine animals, A new brackish-water planktonic co- lectin or agglutinin possesses defense properties pepod belonging to the family Cyclopidae of the against other marine animals. We examined the Copepoda, Apocyclops ramkhamhaengi sp. nov., lectins toward hemagglutinating activity of human is described from a culture originating off Chang and animal erythrocytes, sugar-binding specifici- I., Trat Province, Thailand. The new species is ties, and effect of heat and divalent cations re- the first record of this genus from Thailand and quirement to elucidate the biochemical properties the 4th species of the genus from Asia. Antenna of the sponge lectins. The preliminary screening 2 with an endopod of 2 segments separates for the hemagglutinating activity in 48 marine A. ramkhamhaengi sp. nov. from A. dengizicus sponges was carried out by extraction of the fresh (Lepeshkin). A posterolateral, rounded projec- sponges by 0.85% NaCl. It was found that the tion of pediger 4, and a simple line of spinules protein extracts from nine sponges’ species gave ventrally on pediger 5 separates A. ramkham- more high agglutinating activity against trypsin or haengi sp. nov. from A. royi (Lindberg). A slight papain treated human erythrocytes more than osterolateral projection on pediger 3 and a se- normal erythrocytes. However, the protein ex- ries of ridges dorsally on the basis of maxilla 2 tracts obtained from three species of sponges , are unique for A. ramkhamhaengi sp. nov., and i.e., Chondrilla australiensis, Hyrtios erecta and separate it from A. borneoensis Lindberg. Callyspongia (Euplacella) joubini contained high hemagglutinin specific activity to normal human A erythrocytes with titers approximately at 12,049,

1,965 and 1,932 titer/mg protein respectively. Biodiversity of marine invertebrates Results of sugar-binding specificities assays dwelling in the coral reefs along performed with several carbohydrates and gly- coproteins showed that the lectin from C. aus- the gulf of Thailand traliensis, H. erecta and C. (Euplacella) joubini specifically bound glycoprotein, porcine stomach Sumaitt Putchakarn mucin and fetuin more than mono- or oligosac- Institute of Marine Science, Burapha University, charide. The hemagglutinating activity of lectin from C. australiensis and H. erecta was stabilized Bangsaen, Chonburi 20130 Thailand at 25-50°C and C. (Euplacella) joubini stabilized at 25-35°C. The activity of lectin from H.erecta Abstract could be enhanced by magnesium ion. The Gulf of Thailand is located between latitudes 50 00/ and 130 30/ N and longitudes Fatty acid compositions in 990 00/ and 1060 00/ E, and constitutes a por- Thai marine sponges tion of the shallow Sunda shelf which opens to (order Hadromerida) the South China Sea. The Gulf is approximately 720 kilometers in length, with a coastline of 2,900 Rawiwan Watanadilok, Piyawan Srivilas and kilometers and its maximum depth of about 84 Kanpitcha Jaidee meters. The Gulf locates in the Indo-Malayan sub-region of Indo-west Pacific Zoogeographic Institute of Marine Science, Burapha University, Region, which may contain a very high diversity Chonburi, 20131, Thailand. of marine animals.The Gulf also provides many marine habitats especially coral reefs which dis- Abstract tributes through the Gulf. This paper shows some Seven specimens of marine sponges marine invertebrate diversity dwelling in the coral (Order Hadromerida; Family Spirastrellidae, reefs along the Gulf of Thailand and discusses Tethyidae and Clionaidae) were analyzed for on their needs for fulfill the marine biodiversity their fatty acid compositions. The predominant knowledge through the phyla from the previous compositions were 16:0, 20:5n3 and 22:6n3. surveys and checklist. Most of them were saturated (22.53-36.41%) The field surveys were conducted from and polyunsaturated (19.38-55.54%) fatty ac- 40 sites of 22 islands along the coast of Chon- ids. The essential fatty acids [eicosapentaenoic buri province, over the period January 1997 to acid; EPA, (20:5n3) and docosahexaenoic acid; March 2002. The investigations were carried DHA (22:6n3)] were found at the highest in out by skin and SCUBA diving during daytime. non-polar lipid of Spirastrella sp. (KODA-13; The specimens were collected by random sam- 11.85±0.35%) and Spirastrella solida (LSNG- pling from reef flat toward on outer reef in the 07; 24.36±0.52%). sampling sites. Out of specimens, 428 species were classified from 15 phyla as follow: Porifera 54 species, Cnidaria 151 species, Ctenophora 1 species, Platyhelminth 8 species, Nemertea

2 species, Annelida 21 species, Siphuncula 1 Chonburi 20131, Thailand species, Echiura 1 species, Entoprocta 1 spe- bInstituto de Ciências Biomédicas de Abel cies, Phoronida 1 species, Mollusca 63 species, Salazar, Universidade do Porto, 4099-003 Arthropoda 21 species, Ectoprocta 3 species, Echinodermata 71 species, and Chordata, Uro- Porto, Portugal chordata 32 species. cCentro de Estudos de Química Orgânica, Fitoquímica e Farmacologia da Universidade A comprehensive review of the literature and field surveys on echinoderm recorded in Thai do Porto (CEQOFFUP), Faculdade de waters was carried out. It was found that the Farmácia, 4050-047 Porto, Portugal total numbers of species are 381 species, includ- ing 39 crinoids, 69 asteroids, 112 ophiuroids, 67 Abstract echinoids and 94 holothurians. There are 14 type The ethyl acetate extracts of twenty five species of echinoderms inhabiting in Thai waters species of marine sponges collected from the and four of them are endemic. The Andaman Gulf of Thailand were screened for anticancer Sea has a greater diversity of echinoderms than activity. The results showed that two sponge ex- those in the Gulf of Thailand. Most echinoderm tracts of Hyrtios erecta and Oceanapia sagittaria recorded is of shallow waters while deepwater presented the activity against the human cancer ones are remained relatively unexplored. cell lines. The ethyl acetate extract of the sponge O. sagittaria was found to contain seven known For the two case studies and other compounds, kuanoniamines A and C, together surveys, we found that the distribution of the with 24-methylcholestanol, p-hydroxybenzalde- dominant species of marine invertebrates dwell- hyde, p-hydroxybenzoic acid, phenylacetic acid ing in coral reefs along the Gulf of Thailand and 3-formylindole. Kuanoniamines A and C ex- depended on environment habits, such as water hibited in vitro against the breast cancer cell line turbidity, seasonal monsoon. Moreover, we found (MCF-7) and non-small cell lung cancer cell line that more surveys, we had high opportunity to (NCI-H460) with GI50 value of 0.12, 0.91 AM and find out new species or new recorded marine 0.81, 21.50 PM respectively. The extract of H. invertebrates for Thai Waters. Therefore, ma- erecta is undergoing our further investigation. rine invertebrates in Thai waters have still waited and needed for exploration in order to fulfill the Fatty acid compositions of Thai knowledge of marine invertebrates biodiversity marine sponge Clanthria reinwardi for the region. from four different locations Anticancer activity in ethyl acetate extracts from Thai marine sponges Piyawan Srivilas and Rawiwan Watanadilok Institute of Marine Science. Burapha University, Rawiwan Watanadiloka*, Anake Kijjoab, Maria São José Nascimentoc, Madalena M.M. Pintoc Thailand aInstitute of Marine Science, Burapha University,

Abstract effect of heat stability and divalent cations require- Analysis of fatty acid composition from ment and antibacterial activity of protein extract four different locations of Thai sponge Clath- from Hyrtios erecta. The preliminary screening of ria reinwardti was determined by GC-FID. The the hemagglutinating activity in forty eight marine predominant compositions were saturated fatty sponges was carried out by extraction of the fresh acid (16:0; 38.01- 46.79%) and polyunsaturated sponges by 0.85% NaCl. It was found that the fatty acids (18:2n6, 22:6n-3 and 20:5n-3; 26.44- protein extracts from nine sponges species gave 36.84%). The compositions of the essential fatty more high agglutinating activity against trypsin or acids, eicosapentaenoic acid [EPA] and docosa- papain treated human erythrocytes more than hexaenoic acid [DHA], in this sponge collected normal erythrocytes. However, the protein in Hy- from four locations were compared. The results rtios erecta contained high hemagglutinin specific from Chonburi, Chanthaburi and Trat showed a activity against normal human A erythrocytes to similar tendency, whereas the result from Rayong be 1,965 titer/mg protein. displayed in difference Sugar-binding specificities assays were Biological properties of lectins in performed with several carbohydrates and glyco- marine sponge Hytrios erecta from proteins. The result showed that the lectin from H. erecta specifically bound glycoprotein, porcine Chonburi Province, Thailand stomach mucin, bovine submaxiilary mucin and fetuin more than mono- or oligosaccharide. The Janjarus Watanachote1, hemagglutinating activity of lectin from H. erecta Chutiwan Dechsakulwatana1 and was stabilized at 4-50°C. The lectin activity or hemagglutinating activity from H. erecta could Suriyan Tunkijjanukij2,3 be enhanced by magnesium ion. In addition, H. 1Institute of Marine Science, Burapha University erecta (52.09 g protein) exhibited strong anti- bacterial effect against all tested strains of Vibrio , Chonburi, 20131, Thailand, spp. and gram positive bacteria Staphylococus 2Department of Marine Science, Faculty of aureus ranging from 26-76%. Moreover, the Fisheries, Kasetsart University, Bangkok 10900, protein extract from supernatant and bacterial cells of bacterial associate with H. erecta ex- Thailand. hibited antibacterial effect against V. harveyi, V. 3Center for Agricultural Biotechnology, mimicus and S. aureus ranging from 19-85%. Kasetsart University, Kamphaeng Saen Campus, Nakorn Pathom 73140, Thailand. Abstract In some invertebrate marine animals, lectin or agglutinin possesses defense property. Here, for elucidating the biochemical properties of the sponge lectins, we examined the lectins toward hemagglutinating activity of human and animal erythrocytes, sugar-binding specificities,

Antibacterial and cytotoxic P-388, KB, MCF-7, Lu-1, and/or ASK at < potentiality of sponge extracts and 4 to 15 µg ml-1. The extracts promising result theirs associated bacteria collected s from sponges were Mycale(Zygomycale) parishii, Raspailia(Raspaixilla) sp. new, and Tethya from Chang Islands and seychellensis. In addition the promising bacteri Kood Islands, Thailand a were identified as Pseudomonas spp., Vibrio sp., and Pseudoalteromonas luteoviolacea. The Chutiwan Dechsakulwatana1, se promising results certainly deserve attention i Pawinee Piyachaturawat2, Vichai Reutrakul2, n further study on analysis to identify the active constituents. and Sumaitt Putchakarn1 1Institute of Marine Science, Burapha Study on actinomycetes soluble University, Chonburi 20131, Thailand : pigments for suitable application [email protected] Rattanaporn Srivibool1 and 2Faculty of Science, Mahidol University, Udomluck Thitirakpanich2 1 Institute of Marine Science, Bangkok 10400, Thailand, Burapha University, Chonburi 20131 2) Faculty of Science, Burapha University, Abstract In an attempt to investigate antibacterial Chonburi 20131 and cytotoxic potentiality of sponge extracts an d theirs associated bacteria, the 44 sponges w Abstract ere collected from Chang Islands and Kood Isla Actinomycetes are gram positive bac- nds from the eastern coast of Thailand and 551 teria belonging to the class Actinobacteria which bacterial strains were isolated. These sponge is notably a rich source of biologically active methanol extracts and culturable bacteria were metabolites. Apart from antibiotics for pharma- tested against with 6 test bacteria. The results ceutical and agrichemical uses, low molecular indicated that 14 sponge methanol extracts weight enzyme inhibitors, immunomodifier and and 25 bacterial strains showed antibacteria enzymes for use in industrial applications, ac- l activity against test gram positive bacteria; Bacillus tinomycetes can produce various kinds of pig- subtilis ATCC6633 or Micrococcus luteus ATCC9 ments and soluble pigments. In our study, 284 341 while only 5 samples showed activity against actinomycetes were isolated from coastal areas Pseudomonas aeruginosa ATCC 27853, Vibrio. in Rayong and Chonburi Provinces , Thailand. alginoliticus or Escherichia coli ATCC 25922. The Most of 284 actinomycete isolates could produce evaluation of cytotoxic potentiality of bacterial spore mass in various colors: white, grey, brown, extracts were performed by using MTT assay red, pink, yellow, light yellow, yellow brown, gray- with 6 tumor/cancer cell lines. The promising ish green, and some produced soluble pigments active strains were cultured and partially extracted in yellow, violet, red, brown, green, light brown, with methanol:chloroform(2:1) and ethyl acetate. From 7 active bacterial extracts tested, 3 extracts presenting the most promising activity with

grayish green. Nine samples of crude pigment productivity of volatile acids and high productivity extracts were taken to test for toxicity by using of succinic acid and malic acid. And some Brine Shrimp Bioassay, the results revealed that strains of marine yeast demonstrated high one crude sample was toxic to the brine shrimp. productivity of 2-phenethyl alcohol which is a For suitable application, all the pigments could be beneficial property for white wine improving used for different purposes, more toxic pigments quality1. can be used for fine fabric dyes and non toxic pigments will be investigated further if can be A total of 7 surface seawater and 7 used for food dye. sediment samples from 1.0 -21.8 depth were taken to the laboratory of the Institute of Marin Isolation of marine yeasts from e Science,Burapha University to isolate marine sea water and marine sediments yeasts. One gram of each freezed-dry sediment was suspended in 10 ml sterile for alcoholic fermentation natural seawater, vortexed for 1 minute and setting for 15 minutes. Three isolation media Rattanaporn Srivibool1 Udomluck Thitirakpanich 2 were used for the isolation: Yeast Extract and and Yuji Teramoto3 Malt Extract Agar, Potato Dextrose Agar Agar and Czapex-Dox Agar. All isolation media wi 1Institute of Marine Science, Burapha University, ll be prepared by adding 50% of sterile natural Chonburi. 20131. Thailand. seawater. To prevent growth of other bacteria some antibiotics were used. One hundred 2Faculty of Science, Burapha University. microlitres each of the supernatant samples w Chonburi. 20131. Thailand. ere inoculated to the isolation medium plates in triplicate, including of sediment streak of each 3Faculty of Biotechnology and Life Science, sample in each medium. Seawater samples, Sojo University, Ikeda 4-22-1, Kumamoto 5 ml each, were filtered through 0.22 µm cellulose nitrate filter paper and placed on 860-0082. Japan the isolation plates. All isolation plates were incubated at 35o C for 3-7 days before Abstract observation. The yeast colonies appeared on For thousands of years that people in the isolation plates were then purified and the ancient know how to bake and how to preserved both at 4 o C and freezed dry. All of ferment alcoholic beverages from yeasts. At the yeast isolates were examined for present, yeasts still have been taking an morphological and biochemical study, including important role in alcoholic production, including for further study of molecular identification and alcoholic beverage. To improve better quality the study of alcohol with aromatic compound of beer, wine and other alcoholic production, production. the improving yeast strains is as important as the quality characteristics of the grape or fruit varieties. There was a report, some yeast strains isolated from marine demonstrated low

Species diversity of marine Identification of Vibrio sp. cellulose sponges dwelling in coral reefs in utilizing symbiont bacteria of Had Khanom – Mo Ko Thale Tai bloody cockle National Park, Nakhon Si Thammar (Tegillarca granosa) by 16S rRNA at province, Thailand sequences Sumaitt Putchakarn Kasom Roongcharoen1 , Chuta Boonphakdee2, Institute of Marine Science, Burapha University, Thanomsak Boonphakdee3, Chutiwan Dechsak Bangsaen, Chonburi 20130 Thailand ulwatana4 and Kashane Chalermwat3 Abstract 1Graduate School of Environmental Science, The species diversity of demosponges Faculty of Science, Burapha University, dwelling in the coral reefs at Had Khanom–Mo Ko Bangsaen, Chon Buri, 20131 Thailand Thale Tai National Park, the southern of the Gulf 2Department of Biology, Faculty of Science, of Thailand was investigated, with field surveys Bangsaen, Chon Buri, 20131 Thailand undertaken at 14 sites in November 2006 and 3Department of Aquatic Science, Faculty of May 2007 using SCUBA and random observa- Science, Bangsaen, Chon Buri, 20131 Thailand tion. 47 species of demosponges from 10 orders, 4Institute of Marine Science, Burapa University, 24 families and 34 genera were recorded. Order Bangsaen, Chon Buri, 20131 Thailand Haplosclerida including 15 species was the most abundance species followed with 9 species of Abstract Poecilosclerida and 6 species of Dictyoceratida In this study, we aim to identify symbiont respectively. The massive sponge was the most bacteria of bloody cockle (Tegillarca granosa), dominant growth form of the study area. The which are able to digest/assimilate cellulose, most abundant and common sponges in this the typical terrestial organic matter. DNA was area are: Oceanapia sagittaria; Neopetrosia sp. separately prepared from 12 cellulose agar “blue”, Xestospongia testudinaria; and Haliclona bacterial isolates which were isolated from (Gellius) cymaeformis. Most species are common digestive tract of the bloody cockle, and their representatives of the Indo-Pacific fauna found 16S rRNA genes were subsequently amplified. throughout the Gul fofThailand. Analysis of the 16S rRNA nucleotide sequences revealed that all 12 bacterial strains were highly homologous to Vibrio specie. These results indicate that symbiont Vibrio specie of bloody cockle is able to produce cellulytic enzyme(s). It implies that this bivalve may plays a role in removal of terrestrial organic matter from anestuary system. This work leads us to unders- tand more about biological interaction between land and sea.

คณะผจู ัดทำรายงานประจำป 2552 สถาบันวทิ ยาศาสตรทางทะเล คณะผจู ัดทำ มุธวุ รรณ ประธานคณะทำงาน นายวรเทพ ธนาพงศสมนึก คณะทำงาน นายภัทรพงศ สวสั ด์พิ ีระ คณะทำงาน นางสาวเสาวภา ทองระอา คณะทำงาน นางสาวแววตา กำบุญเลิศ คณะทำงาน นางเออื้ งนภา ลีโทชวลติ คณะทำงาน นางสาวสพุ รรณี กงั วาลโชคชัย คณะทำงาน นายกำพล ทับพร คณะทำงานและเลขานกุ าร นางสาวเบญจวรรณ จันทรด ำ คณะทำงานและผูช ว ยเลขานุการ นางสาวปาณสิ รา ออกแบบและพิมพ เกท็ กูดครเี อชั่น โทร. 038-443202 แฟกซ 038-444931