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Fungus Populations M Marine Waters and Coastal Sands of the Hawaiian, Line, and Phoenix Islands' CAROL WRIGHT STEE LE2

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Fungus Populations M Marine Waters and Coastal Sands of the Hawaiian, Line, and Phoenix Islands' CAROL WRIGHT STEE LE2 Fungus Populations m Marine Waters and Coastal Sands of the Hawaiian, Line, and Phoenix Islands' CAROL WRIGHT STEE LE2 ABSTRACT: Saprophytic and facultative parasitic fungi present in the coastal waters and adjacent pelagic areas of the Hawaiian Islands, and in coastal sands of the H awaiian, Line, and Phoenix islands, were isolated by plating methods. Isolates from all areas sampled indicate that abundant and varied fungus popu­ lations do exist in these environments. The number of fungi obtained from the inshore neritic zone was seven times that obtained from the oceanic zone. The fungus Anreobasidium pltllttla11S ( De Bary) Arnaud was isolated repeatedly from oceanic waters. A comparison is made between the genera and the average number of isolates per liter of water known from the Atlantic Ocean with those found in this study of the Pacific Ocean. The number of fungi isolated from sand samples of the different islands ranged from 2 to 1,600 per gram. Species diversity was evident throughout the samples. The leeward Hawaiian islands had a higher aver­ age number of isolates per gram than any other island group. In conclusion the problems of defining a marine fungus are discussed. OCEA NIC AREAS in different parts of the world also are very limited. These include a few rec­ have been shown to be habitats for marine ords of higher fungi collected in the Marshall fungi (Johnson and Sparrow, 1961) . Investi­ Islands (Rogers, 1947) , the Society Islands gators, however, have usually concentrated on (Olive, 1957, 1958) , and Raroia in the Tua­ particular groups of fungi by use of selective motu Archipelago (Cooke, 1961); Phyco­ isolation methods ( Barghoorn and Linder, mycetes recovered by plating soils of the atolls 1944; Moore and Meyers, 1959; Jones, 1962) . of Bikini, Eniwetok, Rongerik, and Ronggelap Only one extensive analysis of marine waters (Sparrow, 1948); and Ascomycetes and Fungi for a general fungus population is known, and Imperfecti from dung and soil samples col­ it was made in the northwestern subtropical lected by Olive in the Society Islands (Peter­ Atlantic Ocean (Roth et al., 1964) . References sen, 1960) . Consequently, as Cooke points out, to the occurrence of fungi in the Pacific Ocean the geographic distribution of fungi occurring are found (1 ) as incidental to studies of bac­ on the islands of the Pacific is poorly known. teria in marine water (ZoBell, 1946) ; (2) in Although studies have been initiated on the studies of specialized fungi such as lignicolous soils of the Hawaiian Islands ( Baker, 1964) fungi (Cribb and Cribb, 1955 , 1956, 1960; and the phyllosphere (Marsh, 1965) , no study Kohlmeyer, 1960; Meyers and Reynolds, 1960) has been made of fung i occurring in marine and those on algae (Cribb and Cribb, 1955 , waters and intertid al environments of these 1956, 1960); and (3) in studies of particular islands or elsewhere in the central Pacific. This kinds of fungi , e.g., Phycomycetes in Japanese investigation was undertaken to determine the waters (Kobayashi, 1953) and path ogenic spe­ occurrence and distribution of the saprophytic cies (Van Uden and Castelo Branco, 1961 ) . and facultative parasitic fungi which constitute Reports of fungi from terrestrial environments the fungal populations of these habitats. of islands in the central and southern Pacific 1 Prepared from a thesis submitted in partial ful ­ MATERIALS AND METHODS fillment of requireme nts for the Master of Science degree at the University of H awaii. Collection 2 Department of Botany, San D iego State College, San Di ego, Californ ia, 92115. Manuscript received Isolations for fungi were made from 59 June 20, 1966. water samples and 50 sand samples collected 317 318 PACIFIC SCIENCE, Vol. XXI, July 1967 from inshore areas of the Hawaiian Islands and the millipore filter method (Roth et al., and adjacent pelagic environments. Seventeen 1964) . sand samples were taken from the Phoenix and Standard materials and procedures were used Line islands. The water samples were taken in the pour-plate method. Amounts plated for from four zones: the surf or spray zone; the the water samples ranged from 0.5 ml to 5.0 inshore neritic zone, 2-200 m from shore; the ml. Dilutions of 1: 10 to 1: 100,000 were used offshore neritic zone, 300 m to 2 km from for sand samples. Some samples were plated shore; and the oceanic zone, 2 km or more off upon return to the laboratory; because of the shore. Of these water samples 56 were taken distance between most sampling sites and the at the surface and 3 were taken at depths laboratory, however, most of the plating was down to 600 m. The sand samples were done 24 to 48 hours after collection. All sam­ taken from a depth of 2 inches in the supra­ ples were kept under refrigeration until plated. tidal, intertidal, and subtidal zones as delimited A control plate, uninoculated, was set up for by Hedgpeth ( 1957) . The sample sites were each medium for every plating. Plates were selected to include a variety of shore environ­ also exposed to the air during plating opera­ ments: leeward, windward, or areas of special tions to determine the level of air contamina­ interest (e.g., South Point, Hawaii, the south­ tion in the laboratory. ernmost point in the Hawaiian Islands; W ai­ Materials for the millipore filter method kiki Beach, probably the most frequently used included sterile cellulose-ester membranes with beach on Oahu; and Midw ay Island, the north­ 0.45 !! porosity (Millipore Filter Corp., Bed­ ernmost inhabited island of the Hawaiian island ford, Massachusetts) . Th e samples were run chain.) Other samples were obtained as oppor­ through the millipore filter apparatus using a tunity offered : those from Kure Island, the vacuum pump. The membranes with retained Phoenix Islands, the Line Islands, and the fungal elements were cultured on selective me­ open ocean. Figure 1 gives the geographical dia in presterilized pastic petri dishes. The location of the water and sand collection sites. amount put through the filter ranged from The surface water samples were collected in 100 ml to 600 ml per sample. Controls were sterile 600-ml glass bottles with plastic screw included for testing agar sterility and air con­ caps. The closed bottles were immersed in the tamination. water, then opened and allowed to fill with Several kinds of media were used: sodium water, closed underwater and brought to the caseinate agar (BBL 01-549, Fred and Waks­ surface. The depth samples were taken by send­ man, 1928, modified by Potter, 1957), Roth's . ing a plastic water sampler of the van Dorn isolation medium (Roth et al., 1964), Fell's ( 1956) type to the designated depths. After yeast agar (Fell et al., 1960) and mycobiotic the sampler was brought to the deck of the agar ( DIFCO 0689-02) . All media except the ship, a sterile 600-ml bottle was filled with mycobiotic agar were made with sea water col­ water from the sampler. lected at the sample sites. Bacterial growth was All sand samples were collected with sterile controlled by the incorporation of 0.05% chlor­ implements and placed in sterile 100-ml jars amphenicol (Chloromycetin, Steri-Vial N o. 65, with plastic screw caps or in sterile poly­ Parke, Davis and Co.) . Dilutions were made ethylene bags (NASCO Whirl-Pak, Hydro Prod­ with McIlvaine's buffer solution, pH 7.0 ucts Co., San Diego, California) . (Machlis and Torrey, 1956). The mycobiotic Salinity measurements were obtained by use agar was made with 250 ml sea water and of Quantabs S051 (Linayer Corp., Detroit, 750 ml of distilled water in order to retain Michigan) . Temperature was determined by the selectivity of this medium in which chloro­ standard centigrade thermometer, and depth mycetin is already incorporated. by standard oceanographic methods. The plates were placed in paper bags, sealed with adhesive tape, and incubated at 20°_ Isolation 24°C. Pour-plates were incubated for three Two principle means of isolation were em­ weeks and the millipore filter plates for 10 ployed: the pour-plate method (Salle, 1954) days at room temperature (200-24°C). Marine Fungi from Central Pacific-STEELE 319 .,. 110' CENT RAL PACIFIC OC EA N 160" so- '"'"i ~ H IV K i"~ . ""A I. •M L I<>HE"'E. lEU f "i:l fv LISlA SKll ~LAY ANI . I I---~ .- ER NNAC Es () RENe FRO' n 5"0"1.5 ' NE KERl NIHO , ~AI P , ~J., pu' U' .....A ~ fi .; " 0 OK" In '" m~ .:- "A Ul KA. foLAwt 1 20' N' ~X 1-1 WAil . - --_.. i: ""NS e-. -$AN O l - - - 10" '0 ' I A~R K'N( EF -II '-0 . PALMYRA! - - -- I--- - I---I--- . i's "~TON ' r-- \. ANNING l . I - - I--- -- CH 15TM 5 1. " -I-- -- - - U' --- e-- - - - . - CI -;j;Q <\.' ii ,-- -- O. ER , 0" 0" 0 JARVIS l _. _- _ . 1-- -- ' -- -- -~- I--- - I- f---. - I--- - _._. ._._~ AN. .L - _. - --_. " I--- --- .- --- . ~N \ . IS 1- - --- - - - 1'1"1 TON, --_.. - 1-- -- 0 -"". ">T'e..,' ~ 0 _~--~_f~-__l 0 RNiE IC? .. N,. -- , ;.~, L , - <lYONE "Al """ - - ( .: _.. ..•_- .. .._- ---- ,- STA BUCKI. - --- - -_. __. " _...- ._.. - .. _-- ---- -- II> 1-'\SL/Wo -- . ' I ~\l '---- _ . • A AfUl TONG> EVA - - -- <f I--- - NU~ = ~H "iO' I ""'" IHEllO" ,0- TATUn: l5O"~JI-I ~WE5T - 'L , - FIG. 1. General geographical location of marine water and coastal sand collection sites. X , Marin e water collection sites; 0 , coastal sand collection sites. 320 PACIFIC SCIENCE, Vol. XXI, July 1967 Oth er methods of isolation, such as baiting successfully were Czapek-Dox agar (DIFCO (Johnson and Sparrow, 1961) , spread-plate 0339-01) and "V-S" juice agar (Wickerham technique (Bu ck and Cleverdon, 1960) and et aI., 1946).
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