Fungi in and Near Streams Carrying Acid Mine-Drainage1 Wm
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FUNGI IN AND NEAR STREAMS CARRYING ACID MINE-DRAINAGE1 WM. BRIDGE COOKE, 1135 Wilshirc Ct., Cincinnati, Ohio 45230 Abstract. From streams carrying acid mine-drainage and from adjacent soils in 3 areas in Ohio and West Virginia, 189 species of fungi, including yeast like as well as filamentous types, were isolated using agar pour plates and shaken flask cultures. The more important members of this population were members of the Fungi Im- perfecti, 25% of these yeast like and the remainder filamentous fungi. Species with phialidic conidium production represented 80% of the filamentous fungi. Larger numbers of species and colonies were recovered during the mid-autumn sampling period. OHIO J. SCI. 76(5): 231, 1976 Continuing studies of the biology of fungi, previously indicated only by total acid mine-drainage carrying streams in numbers for a habitat, a location, a date, the soft coal producing regions of Ohio or the project, be listed. and West Virginia by units of the organi- zation then known as the Robert A. Taft METHODS Sanitary Engineering Center, and the Sample locations for the Ohio portion of the Ohio Basin Region of the Federal Water study were selected by the writer, who made Pollution Control Administration, indi- all Ohio collections with the help of his wife. Personnel from the Wheeling, W. Va., and the cated a need for information about fungal Elkins, W. Va., laboratories assisted in choos- populations of this habitat. A report on ing sites for the West Virginia samples. The the studies made in 1964 and 1965 was writer made all initial field collections and field published by Cooke (1966). In develop- personnel took the second and third sets using bus express for overnight shipment to Cin- ing a catalogue of the fungi of Ohio in cinnati. Data on pH, total Fe in ppm, Fe++ streams carrying acid-mine drainage, it in the presence of Fe+++ in ppm, and number of seems appropriate that the species of species and colonies of fungi in each sample on each sampling date were given in Cooke (1966). Manuscript received December 17, 1975 and Techniques of obtaining this information were in revised form April 20, 1976 (#75-74). given briefly in that report. Sample Locations — Ohio Series (O) 1. Sandy Run at King Hollow Bridge, off SR-278, Vinton Co., Ohio 2. Sandy Run below mine at mouth of Coal Hollow, Vinton Co., Ohio. 3. Sandy Run above mine at mouth of Coal Hollow, Vinton Co., Ohio. 4. Sunday Creek at junction of Township Road 291 and SR-13, Perry Co., Ohio. 5. Sunday Creek near Hemlock above bridge on SR-165, Perry Co., Ohio. 6. Bricked in spring on hillside berm of SR-216 near Murray City, Hocking Co., Ohio. 7. Creek from spring in ochre "cave", east of New Straitsville, along SR-216, Hocking Co., Ohio. 8. Crumley's Farm Creek, off US-33, near Groveport, Franklin Co., Ohio. 9. Ash Cave Falls, Ash Cave State Park, Hocking Co., Ohio. 9a. Wheeling Creek, Belmont Co., Ohio, just above Belmont Co. road No. 10, bridge below active coal mine. Sample Locations — Morgantown Series (M) 1. Fifty yards above mouth of Scott's Run, of W. Va. Rte. 100, Monongalia Co. 2. Decker's Creek below power plant, about one mile above mouth, Morgantown. 3. Booth Creek, 2.1 mi. above Weirton Steel Co. coal mine preparation plant. 4. Booth Creek at WTeirton Steel Co. coal mine preparation plant (abandoned), between gob piles and settling basin. 5. Lake Lynn on Cheat River at boat docks below State Park Lodge. 6. Near mouth of Robinson Run, 1.7 mi. n. of US-19 on right side of road. 7. Monongahela River, west bank, 2.5 mi. n. of US-19 bridge below Morgantown. 8. Monongahela River, east bank, 0.5 mi. S. of Morgantown Dam. 231 232 WM. BRIDGE COOKE Vol. 76 Sample Locations — Elkins Series (E) 1. Station R-l, near mouth of Roaring Creek, 100 ft. above bridge. 2. Station G-l, near mouth of Grassy Run, at old railroad bridge. 3. Station GT-6-1, Norton Mine # 15 drainway. 4. Station RT-5-3, Pond on White's Run, top of strip pit. 5. Station R-3, Roaring Creek at Coalton. 6. Station TR-6-17, Kittel's Run, including samples of material from aluminum and iron "travertine" deposits built up on leaf litter at springs. 7. Station R-C Special (R-9), Roaring Creek. 8. Station RT-6 Basin—Pond in strip pit by Coalton Dump. 9. Station RT-10-1. Brooks Run, tributary of Roaring Creek. In table 1, habitats are given for each species high-water mark where the habitat was rarely but no attempt has been made to indicate the affected by the stream water, (see table 1). date or dates on which a species was recovered, and the locations have been lumped for each Pour plates were prepared as in Cooke (1963). area. The three basic areas sampled were: For these, two media were used in parallel: Ohio, including stations on the Hocking River neopeptone-dextrose-rose bengal-Aureomycin, and upper Racoon Creek; West Virginia, in- and neopeptone-dextrose-Aureomycin agars, the cluding the Morgantown area, Monongalia Co., latter acidified to approximately pH 5.0. The with stations in the Monongahela River basin; third and fourth media were YNB-1% dextrose, the Elkins area, Randolph Co., W. Va., with and YNB-20% dextrose broths, prepared at stations in the Cheat River basin. For each room temperature and filter sterilized accord- basic area 8 or 9 locations were chosen ing to Cooke (1963). More discrete colonies as indicated above. At each location 4 appeared in the medium including rose bengal, habitats were sampled: flowing water (W) and yeasts grew better in broth media in in the stream in which spores were carried pas- shaken culture. Elements of the total popula- sively, or colonies of fungi could be actively tion, so far as it was obtained, appeared on growing; sediments (S) on the bottom of the each medium but no single medium gave a stream at arm's length from the shore from complete picture of either yeasts or filamentous which stream water could not be excluded but fungi. In a fifth technique, Oomycetes were in which fungal colonies could be present as obtained only when portions of samples were members of the benthos community; stream incubated with hemp seeds (Cooke, 1963). bank (B) soil taken at the interface of the Species were identified by the author based on stream and the bank so that the habitat was either his interpretation of monographic or always affected by the stream water; and up- mycologistic treatments, or by comparison land (U) soil usually taken above the obvious with known strains or subcultures. TABLE 1 Systematic list of Fungi Recovered from Acid Mine Drainage Sites in Ohio and West Virginia. No. of Isolations! Species Total West Virginia Ohio Samples* Morgantown Samples* Elkins Samples* Eumycola 2827 Mastigomycolina Oomycetes 11 Saprolegniales Saprolegniaceae Achlya spp.** 6 B-l.U-1. Aphanomyces sp.** 1 W-l. Diclyuchus sp. 1 S-1. Saprolegnia spp. 3 W-2.B-1. Zygomycotina Zygomycetes 252 Mucorales Mucoraceae Absidia cylindrospora 1 S-1. Absidia glauca 3 S-1,B-1 S-1. Actinomucor elegans 1 S-1. Gongronella butleri 5 S-J.U-l S-l.B-2. Mucor spp.** 102 34 40 Mucor alternans 2 S-1. U-l. Mucor hiemalis 66 34 28 No. 5 FUNGI IN ACID MINE-DRAINAGE 233 TABLE 1. Continued. No. of Isolations"!' Species Total West Virginia Ohio Samples* Morgantown Samples* Elkins Samples* Mucor racemosus 2 B-l. U-1. Rhizopus spp. 20 W-J,S-5,B- 5. S 1,B 1. W-l,B-2,U-4. Rhizopus arrhizus 1 W- 1. Rhizopus cliinensis Series 3 \Y- l.U-1. B-l. Zygorliyuchus moelleri 36 \Y 1.S-6.B (i.U-5. \Y 1,S 2,B-4,U-3. S-2.B-2.U-4. Mortierellaceae Morlierella angulisporus 18 \Y- 1,S 2,B-2,U-1. \Y-2,S-2,B-l,U-3. B-l,S-l,U-2. Cunninghamellaceae Cunninghamella elegans 1 S-1. Piploce phalidaccac Synce phalastrum racemosum 1 1 Ascomycotina 21 Saccharomycelaceae Endomycopsis burlonii—see Trichosporon behrendii Hansenula salurnus 1 S-1. Pichia guiltiermondii 7 S-1,B-l,U-1. B 2. B-l,U-1. Saccharomyces sp. 2 B-2. Saccharomyces cerevisiae 10 W-l.B-2. W-l.S-2. S-2,B-1,U-1. Saccharomyces rosei 1 B-l. Plectomyceles Eurotiales Melanosporaceae Chaetomium globosum 1 YV-1. Sordariaceae Pleosporales S porormiaceae Preussia multispora 1 B-l. Basidiomycolina*** Teliomyceles Ustilaginales Rhodotorula glulinis Deuteromycotina 2542 Coelomycetes Sphaeropsidales 152 Sphaeropsidaceae Chaetomella sp. 3 B-l.U-2. Coniothyrium sp. 4 S-1, 1. Coniothyrium fuckelii 1 1 Cytospora sp. 1 1 Phoma spp. 108 +30 +24 +30 Phoma herbarum 31 W-4,S-5,B-7,U-2. W-2,S-3,B-4,U-2. W-2. Selenophoma sp. 1 1 Septoria sp. 1 1 Sphaeronema spinella 1 1 Blastomycetes Cryptococcales Cryptococcaceae Candida curvata 26 W-l,S-3,B-3,U-2. S-2,B-2,U-1. W-2,S-3,B-3,U-4. Candida guiltier mondii 15 W-2.S-1.B-1. W-l,S-3,B-3,U-2. S-1, U-1. Candida humicola 14 S-3,B-2,U-1. S-2,B-4,U-2. Candida intermedia 6 U-1. B-3. B-l,U-1. Candida krusei 37 S-4,B-6,U-4. W-3,S-6,B-4,U-3.