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Cut-lengths of perennial ryegrass leaf-blades influences in vitro fermentation by the anaerobic fungus neocallimastix frontalis.

Jimenez, Hugo R.; Edwards, Joan E.; Sanderson, Ruth; Kingston-Smith, Alison H.; McEwan, Neil R.; Theodorou, Michael K.


Hugo R. Jimenez

Joan E. Edwards

Ruth Sanderson

Alison H. Kingston-Smith

Neil R. McEwan

Michael K. Theodorou


Anaerobic fungi in the gut of domesticated and wild mammalian herbivores play a key role in the host’s ability to utilize plant biomass. Due to their highly effective ability to enzymatically degrade lignocellulose, anaerobic fungi are biotechnologically interesting. Numerous factors have been shown to affect the ability of anaerobic fungi to break down plant biomass. However, methods to reduce the non-productive lag time in batch cultures and the effect of leaf-blade cut-length and condition on the fungal fermentation are not known. Therefore, experimentation using a novel gas production approach with pre-grown, axenic cultures of Neocallimastix frontalis was performed using both fresh and air-dried perennial ryegrass leaf-blades of different cut-lengths. The methodology adopted removed the lag-phase and demonstrated the digestion of un-autoclaved leaf-blades. Fermentation of leaf-blades of 4.0 cm cut-length produced 18.4% more gas yet retained 11.2% more apparent DM relative to 0.5 cm cut-length leaf-blades. Drying did not affect fermentation by N. frontalis, although an interaction between drying and leaf-blade cut-length was noted. Removal of the lag phase and the use of un-autoclaved substrates are important when considering the biotechnological potential of anaerobic fungi. A hypothesis based upon sporulation at cut surfaces is proposed to describe the experimental results.


JIMENEZ, H.R., EDWARDS, J.E., SANDERSON, R., KINGSTON-SMITH, A.H., MCEWAN, N.R. and THEODOROU, M.K. 2020. Cut-lengths of perennial ryegrass leaf-blades influences in vitro fermentation by the anaerobic fungus neocallimastix frontalis. Microorganisms [online], 8(11): unleashing the hidden potential of anaerobic fungi, article ID 1774. Available from:

Journal Article Type Article
Acceptance Date Nov 4, 2020
Online Publication Date Nov 11, 2020
Publication Date Nov 30, 2020
Deposit Date Nov 12, 2020
Publicly Available Date Nov 12, 2020
Journal Microorganisms
Electronic ISSN 2076-2607
Publisher MDPI
Peer Reviewed Peer Reviewed
Volume 8
Issue 11
Article Number 1774
Keywords Neocallimastigomycota; Fermentation kinetics; Gas production; Perennial ryegrass; Neocallimastix frontalis
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