Members of Dimargaritales and Piptocephalida-
ceae (Zoopagales) are haustorial parasites of other
fungi, especially Mucorales;C. recurvatusPoitras is
the host of choice (Benjamin 1959 ). Species ofMortier-
ella,Umbelopsis,Penicillium, andChaetomiumalso
may be hosts for a few mycoparasites (Benjamin 1959 ,
1979 ) in the laboratory. The optimal isolation and
growth temperature for species ofPiptocephalis is
18 C; reproduction is normal, but growth is slightly
slower.
Cryogenic storage at196 to 80 Cin
ultracold mechanical freezers or in liquid nitro-
gen in special Dewar flasks is possible for nearly
all zygomycotan fungi and may be required for
some groups (especially Entomophthorales).
Fungi can also be maintained in pure axenic
(two-membered for parasites) culture, refriger-
ated after sporulation, and transferred periodi-
cally (every 6–12 months for most taxa but as
often as every 2–3 weeks for some Ento-
mophthorales, e.g.,BasidiobolusandConidio-
bolus). Many of the remaining zygomycetes can
be lyophilized or stored under liquid nitrogen,
sterile distilled water, or sterile mineral oil for
long-term preservation.
Most entomophthoraleans can be isolated
on pure culture on relatively simple media,
such as Sabouraud dextrose agar +1 % yeast
extract, or on a variety of media incorporating
coagulated egg yolk (Papierok and Hajek 1997 ).
Cultures of members of Entomophthorales can-
not be preserved by lyophilization but are gen-
erally readily storable in cryogenic freezers,
preferably at 196 C (immersed in liquid
nitrogen) using 10 % glycerol as cryoprotec-
tant. Humber ( 1994 ,1997b) discusses several
of the problems and techniques of culturing
and preserving entomophthoraleans.
Commercial-scale growth of ento-
mophthoraleans for use in biological control
may not be possible for taxa whose vegetative
stages are naturally protoplastic, such asEnto-
mophagaspp. that are pathogenic for caterpil-
lars, grasshoppers, and locusts. Taxa whose
vegetative growth is walled, such asZoophthora
spp. affecting a very wide range of hosts, are
more tractable subjects for large-scale aerobic
fermentation. The best strategy for the mass
production and use of entomophthoraleans
for insect biocontrol is by the controlled drying
of the mycelium (McCabe and Soper 1985 ),
which when ground into flakes, rehydrated,
and applied produces infective spores in the
midst of the target population. There is no
adequate or practical strategy for the produc-
tion and use of entomophthoralean conidia or
resting spores for biological control.
Entomophthoralean saprobes in soil and
plant detritus, mainly species ofConidiobolus
andBasidiobolus, are usually detected only by
using Drechsler’s ( 1952 ) canopy plating isola-
tion technique. Independent studies on terrico-
lous, saprobic entomophthoraleans in the USA
by C. Drechsler and D. King, and in India by
Srinivasan, Thirumalachar, and Narasimhan
(cited in King 1977 ) suggest that these fungi
may be taxonomically diverse and relatively
abundant wherever they are specifically sought.
The species that have been cultured are
primarily the entomopathogens and species of
ConidiobolusandBasidiobolus; no species of
the Meristacraceae (Meristacrum, Tabano-
myces) or the phytopathogenic generaAncy-
listesand Completoriahave been isolated in
axenic cultures.
Members of Harpellales currently in axenic
culture only consist of a number of taxa from the
hindguts of many types of aquatic insect larvae:
manySmittiumspp. from six families of dip-
teran larvae,Austrosmittiumsp. andTrichozy-
gospora chironomidarumLichtw. from midge
larvae, andCapniomyces stellatusS.W. Peterson
&Lichtw.,Genistelloides hibernus,andSimulio-
myces spica S.W. Peterson & Lichtw. from
winter-emerging stonefly nymphs.
Ordinary transfer procedures will not work
with members of Harpellales because they are
aquatic fungi. Details of the culture methods
and physiology of Harpellales can be found in
Lichtwardt ( 1986 ) and Lichtwardt et al. ( 2001 ).One of the best culture media is dilute brain-heart infu-
sion (one-tenth the normal concentration) agar with
vitamins (one-tenth BHIv) (Lichtwardt 1986 ). For rou-
tine culture maintenance, one-tenth BHIv can be
prepared as slants in screw-capped test tubes. Then
add a small amount of sterile, distilled water (1.5 cm)
at the bottom of the tube when it is upright. WATER IS238 G.L. Benny et al.