Mycopathologia vol. 58, 2, pag. 115 116, 1976
MYCOFLORA OF ACTIVATED SEWAGE SLUDGE U.L. DIENER, G. MORGAN-JONES, W.M. HAGLER, Jr. & N.D. DAVIS Department of Botany and Microbiology, Auburn University Agricultural Experiment Station, Auburn, Alabama 36830
Abstract Thirty-eight species of fungi were identified in pure culture after isolation from activated sewage sludge by serial dilution. Nine species and genera were identified that had not been previously reported. In 1963, Cooke (1) published an excellent laboratory guide on the identification of fungi from polluted water, sewage, and sewage treatment systems; of approximately 30 papers cited only one (2) dealt with fungi from activated sewage sludge. Later (1970), Cooke & Pipes (3) enumerated 47 fungi consisting of 4 genera of yeasts and 33 ,genera of filamentous fungi that had been isolated from activated sludge. This paper reports the mycoflora of anaerobically digested sludge from a residential area in Auburn, Alabama.
days after culturing. Pure cultures were transferred to test tube slants of yeast extract (YE) agar medium containing 5% dextrose, 0.7% yeast extract (Difco), and 0,5~o KH2PO4.
Results Dilutions of 1 : 20,000 and 1 : 200,000 were most suitable for isolating colonies to pure cultures averaging about 25 and 8-10 colonies per plate, respectively. Thirty-eight fungi isolated from sludge and identified to species, wherever possible, are listed in Table 1. Five species of Aspergillus and nine species of Penicillium predominate among the 38 species, most of which are members of the Fungi Imperfecti.
Discussion Materials and methods Samples of sludge were collected from drying beds at the North Auburn sludge plant by removing the top half inch of sludge with a spatula and taking the sample from the next 1-11/z inches. Sludge was placed in sterile 250 ml flasks and taken to the laboratory where serial dilutions were made and plated the next day. Ten g of sludge was added to 200 ml of sterile water and mixed for 6 rain in a Sargent Centrifugal Wet Mill to make a 1 : 20 dilution. Ten ml was added to 90 ml to make a 1 : 200 dilution. Each dilution was shaken 5 min at 100 rpm on a Burrell Wrist - Action shaker. Dilutions of 1 : 1000, 1 : 20,000 and 1 : 200,000 and 1 : 2,000,000 were made in a similar manner. One ml of each of the three highest dilutions was added to each of 3 sterile plates. About 10-12 ml of melted peptone - dextrose agar with rose - bengal and streptomycin (50C) was added to each plate (4). A second set of plates was made with potato dextrose agar acidified to pH 5 with 40% latic acid, Fungal colonies were transferred to plates of potato dextrose agar or Czapek - Dox agar 6 and 14
This list of fungi compares rather closely with that of Cooke & Pipes (3) when it is considered that they did not provide specific identities for all members of Aspergillus, Penicillium, Cladosporium, Fusarium and Phoma isolated. Additional genera and species were: Allereschia crocea, Chrysosporium pannorum, Dactylaria sp., Dactylomyces sp., Dichotomomyces albus, Gliomastix musicola,
Pseudoeurotium zonatum, Talaromyces spiculisporus, and Thamnidium elegans. Five of these genera were, however, cited by Cooke (1) as occurring in sewage systems.
Dactylomyces sp., Dichotomomyces albus, Pseudoeurotium zonatum, and Thamnidium elegans were not previously reported as occurring in this habitat.
Acknowledgment This research was supported in part by funds provided by the Office of Water Research and Technology of the U.S. Department of the Interior through the Water Resources 115
Table 1. Fungi isolated from North Auburn sewage sludge
Research Institute A-030-A 1a.
AUA Culture no.
Fungus
References
876 874 782 A-55 882 A-41 B 861
Allereschia [Petriellidium) crocea Aspergillus brunneo-uniseriatus A. flavus A. fumigatus A. parasiticus A. versieolor Acremonium sp. Chrysosporium pannorum Cladosporium herbarum C. cladosporioides Dae ty laria sp. Dactylomyees sp. Dichotomomyees albus Epieoecum purpuraseens Fusarium moniliforme F oxysporum Geotrichum candidum Gliocladium roseum Gliomastix musicola Mucor hiemalis Paeeilomyces niveus Penieillium brevicompactum P. corylophilum P. cylopium P. diversum P. implicatum P. lanosum P. steckii P. stoloniferum P. variabile Phoma eupyrena P. herbarum Pseudoeurotium zonatum Rhizopus arrhizus Sepedonium sp. Sporothrix sp. Talaromyces spieulisoprus Thamnidium elegans
781 790 859
A-12 779 567 857A A-49 860 A-5 789 867
5 A-33 871 A-3 869 857B 799 A-2 858 791 A-54 780 A-27 A-52 476 873 A-11 870 A-37
116
of
Auburn
University
Project
1. Cooke, W.B. 1963. A laboratory guide to fungi in polluted waters, sewage, and sewage treatment systems. Pub. Health Serv. Pub. 9 9 9 - W P - 1 , USDHEW, Cincinnati, Ohio. 2. Cooke, W.B., and F. Ludzack. 1958. Predacious fungus behavior in activated sludge systems. Sewage Ind, Wastes 30: 1490-1491. 3. Cooke, W.B., and W.O. Pipes. 1970. The occurrence of fungi in activated sludge. Mycopathol. Mycol. Appl. 40: 249-270. 4. Johnson, L.F., and E.A. Curl. 1972. Methods for research on the ecology of soil - borne plant pathogens, p. 196. Burgess Publishing Co., Minneapolis.
MYCOFLORA OF ACTIVATED SEWAGE SLUDGE U.L. DIENER, G. MORGAN-JONES, W.M. HAGLER, Jr. & N.D. DAVIS Department of Botany and Microbiology, Auburn University Agricultural Experiment Station, Auburn, Alabama 36830
Abstract Thirty-eight species of fungi were identified in pure culture after isolation from activated sewage sludge by serial dilution. Nine species and genera were identified that had not been previously reported. In 1963, Cooke (1) published an excellent laboratory guide on the identification of fungi from polluted water, sewage, and sewage treatment systems; of approximately 30 papers cited only one (2) dealt with fungi from activated sewage sludge. Later (1970), Cooke & Pipes (3) enumerated 47 fungi consisting of 4 genera of yeasts and 33 ,genera of filamentous fungi that had been isolated from activated sludge. This paper reports the mycoflora of anaerobically digested sludge from a residential area in Auburn, Alabama.
days after culturing. Pure cultures were transferred to test tube slants of yeast extract (YE) agar medium containing 5% dextrose, 0.7% yeast extract (Difco), and 0,5~o KH2PO4.
Results Dilutions of 1 : 20,000 and 1 : 200,000 were most suitable for isolating colonies to pure cultures averaging about 25 and 8-10 colonies per plate, respectively. Thirty-eight fungi isolated from sludge and identified to species, wherever possible, are listed in Table 1. Five species of Aspergillus and nine species of Penicillium predominate among the 38 species, most of which are members of the Fungi Imperfecti.
Discussion Materials and methods Samples of sludge were collected from drying beds at the North Auburn sludge plant by removing the top half inch of sludge with a spatula and taking the sample from the next 1-11/z inches. Sludge was placed in sterile 250 ml flasks and taken to the laboratory where serial dilutions were made and plated the next day. Ten g of sludge was added to 200 ml of sterile water and mixed for 6 rain in a Sargent Centrifugal Wet Mill to make a 1 : 20 dilution. Ten ml was added to 90 ml to make a 1 : 200 dilution. Each dilution was shaken 5 min at 100 rpm on a Burrell Wrist - Action shaker. Dilutions of 1 : 1000, 1 : 20,000 and 1 : 200,000 and 1 : 2,000,000 were made in a similar manner. One ml of each of the three highest dilutions was added to each of 3 sterile plates. About 10-12 ml of melted peptone - dextrose agar with rose - bengal and streptomycin (50C) was added to each plate (4). A second set of plates was made with potato dextrose agar acidified to pH 5 with 40% latic acid, Fungal colonies were transferred to plates of potato dextrose agar or Czapek - Dox agar 6 and 14
This list of fungi compares rather closely with that of Cooke & Pipes (3) when it is considered that they did not provide specific identities for all members of Aspergillus, Penicillium, Cladosporium, Fusarium and Phoma isolated. Additional genera and species were: Allereschia crocea, Chrysosporium pannorum, Dactylaria sp., Dactylomyces sp., Dichotomomyces albus, Gliomastix musicola,
Pseudoeurotium zonatum, Talaromyces spiculisporus, and Thamnidium elegans. Five of these genera were, however, cited by Cooke (1) as occurring in sewage systems.
Dactylomyces sp., Dichotomomyces albus, Pseudoeurotium zonatum, and Thamnidium elegans were not previously reported as occurring in this habitat.
Acknowledgment This research was supported in part by funds provided by the Office of Water Research and Technology of the U.S. Department of the Interior through the Water Resources 115
Table 1. Fungi isolated from North Auburn sewage sludge
Research Institute A-030-A 1a.
AUA Culture no.
Fungus
References
876 874 782 A-55 882 A-41 B 861
Allereschia [Petriellidium) crocea Aspergillus brunneo-uniseriatus A. flavus A. fumigatus A. parasiticus A. versieolor Acremonium sp. Chrysosporium pannorum Cladosporium herbarum C. cladosporioides Dae ty laria sp. Dactylomyees sp. Dichotomomyees albus Epieoecum purpuraseens Fusarium moniliforme F oxysporum Geotrichum candidum Gliocladium roseum Gliomastix musicola Mucor hiemalis Paeeilomyces niveus Penieillium brevicompactum P. corylophilum P. cylopium P. diversum P. implicatum P. lanosum P. steckii P. stoloniferum P. variabile Phoma eupyrena P. herbarum Pseudoeurotium zonatum Rhizopus arrhizus Sepedonium sp. Sporothrix sp. Talaromyces spieulisoprus Thamnidium elegans
781 790 859
A-12 779 567 857A A-49 860 A-5 789 867
5 A-33 871 A-3 869 857B 799 A-2 858 791 A-54 780 A-27 A-52 476 873 A-11 870 A-37
116
of
Auburn
University
Project
1. Cooke, W.B. 1963. A laboratory guide to fungi in polluted waters, sewage, and sewage treatment systems. Pub. Health Serv. Pub. 9 9 9 - W P - 1 , USDHEW, Cincinnati, Ohio. 2. Cooke, W.B., and F. Ludzack. 1958. Predacious fungus behavior in activated sludge systems. Sewage Ind, Wastes 30: 1490-1491. 3. Cooke, W.B., and W.O. Pipes. 1970. The occurrence of fungi in activated sludge. Mycopathol. Mycol. Appl. 40: 249-270. 4. Johnson, L.F., and E.A. Curl. 1972. Methods for research on the ecology of soil - borne plant pathogens, p. 196. Burgess Publishing Co., Minneapolis.
