Degradation of Li gno cellulose Material by Marine Fungi 317 



of the wrapped ends extended beyond the lip of the flask, the foil 

 covering allowed the material to remain sterile after autoclaving 

 and throughout tlie gi'owth period. 



The flasks of the Manila yeast extract medium were steri- 

 lized at 121 C for twenty minutes. The sterilization process itself 

 had no measurable effect on the physical properties of the 

 Manila, nor was any deleterious effect noted when the sterilized 

 cordage remained in the yeast extract sea water medium for a 

 period of several months and longer. 



The inoculum for the culture vessels was 1 to 2 ml of spore 

 and mycelial suspensions from individual stock cultures, prepared 

 by flooding the slant cultures with sterile sea water. 



Inoculated flasks were incubated at 25 C from five days to 

 as long as several months on an intermittent reciprocating shaker 

 set at 60 strokes/min. Flasks were harvested when extensive 

 fungal growth was evident on the twine. Culture vessels were 

 examined for extent of reproduction, cellulolytic activity of the 

 cell-free filtrate, and tensile strength measurements of the in- 

 fested twine. 



Analyses of cellulolytic activity of the cell-free filtrates have 

 been described previously (1, 2). The Nelson-Somogyi method 

 (6) was used to determine the reducing sugars, RS, of the fungal 

 filtrates and the RS produced during the enzymatic tests. 



Treatment of tlie field-exposed cordage for tensile strength 

 properties was similar to the procedure followed with cordage 

 attacked in uniclonal cultures. Uniform periods of drying and 

 conditioning of the twine were maintained throughout the study. 

 Control pieces were tested regularly along with the material ex- 

 posed in the field to ascertain any possible effect of the drying 

 and conditioning processes on the Manila. Repeated use of such 

 controls has demonstrated that these latter processes do not af- 

 fect the strength readings or the regularity of the results ob- 

 tained. 



A Dillon Model K Universal Tester has been used in the 

 present work. Use of this instrument has permitted strength de- 

 termination on small sections of cordage, i.e., six to eighteen 

 inches long. The Scott Tester, used in earlier studies, required 

 a minimum length of five feet of cordage for each test and since 



