Plant and Soil 235: 2734, 2001.

2001 Kluwer Academic Publishers. Printed in the Netherlands.

27

Effect of chitin waste-based composts produced by two-phase composting

on two oomycete plant pathogens
Cinthia Labrie1 , Philippe Leclerc1 , Nathalie Cte1 , Sebastien Roy1 , Ryszard Brzezinski1 ,
Richard Hogue2 & Carole Beaulieu1,3
1 Centre

dEtude
et de Valorisation de la Diversite Microbienne, Departement de biologie, Universite de
Sherbrooke, Sherbrooke (Quebec), Canada, J1K 2R1. 2 Institut de Recherche et de Developpement en Agroenvironnement, Sainte-Foy (Quebec), Canada, G3P 3W8. 3 Corresponding author
Received 22 June, 2000. Accepted in revised form 4 May 2001

Key words: composting, disease suppression, fatty acids, Gram-positive bacteria, Phytophthora, Pythium

Abstract
Biphasic composts were prepared by first mixing peat moss and sawdust with a nitrogen-rich biomass such as
chitinous waste or cow manure and composting them until termination of the thermophilic phase. These partially
stabilized composts were then amended with shrimp waste inducing a second thermophilic phase. Filter-sterilized
water extracts obtained from two mature biphasic composts (SP2 W2 +S and MPW+S) reduced the growth of two
oomycete plant pathogens, Phytophthora fragariae var. rubi and Pythium ultimum. Both SP2 W2 +S and MPW+S
composts significantly reduce the incidence of cucumber damping-off caused by Pythium ultimum as compared
to a commercial brand of compost made from shrimp waste and peat moss. Hydrolysis products of chitin were
unlikely to be responsible for growth inhibition since no oligomeric forms of chitin were detected in SP2 W2 +S.
The shrimp waste amendment carried out after the first thermophilic phase modified the microbial populations
of biphasic composts. Following the amendment, the proportion of branched-chain microbial fatty acids typical
of Gram-positive bacteria increased considerably suggesting that this group of bacteria became more prevalent
within the total microbial population. These data suggests that the two-phase composting process promotes the
proliferation of Gram-positive bacteria antagonistic to oomycete plant pathogens.
Abbreviations: CFU colony forming units; MPW+S compost made of manure peat moss and sawdust in a 1:1:1
volumetric ratio and amended with one-third volume of shrimp waste after the first thermophilic phase; NHF N-acetylglucosaminidase-hydrolysable fraction; PDA potato dextrose agar; PDB potato dextrose broth; SDS
sodium dodecyl sulfate; SP2 W2 compost made of shrimp waste, peat moss and sawdust in a 1:2:2 volumetric
ratio; SP2 W2 +S compost made of shrimp waste, peat moss and sawdust in a 1:2:2 volumetric ratio and amended
with one-third volume of shrimp waste after the first thermophilic phase
Introduction
Environmental pressure has increased over the last
decade to reduce fungicide utilization in agriculture.
Compost products have been viewed as potential
substitutes for fungicides; several independent studies demonstrated that composts could reduce the incidence of plant diseases (De Ceuster and Hoitink,
FAX No: 1 819 821 8049.

E-mail: carole.beaulieu@courrier.usherb.ca

1999a). For example, strong indications exist that oomycete plant pathogens can be controlled by compost
amendment to plant growth substrate (Ben-Yephet
and Nelson, 1999; Craft and Nelson, 1996; Kim et
al., 1997; Theodore and Toribio, 1995). The pathogen suppressive property of compost-amended media
depends on several factors such as the type of biomass composted, composting process, stability of
the products, application time and compost dose (De
Ceuster and Hoitink, 1999b). Different mechanisms

28
have been proposed to explain the suppression of plant
diseases by composts: induction of systemic acquired
resistance in plants (Zhang et al., 1998), lethal effect
of chemical components on plant pathogens (Cronin
et al., 1996) and stimulation of microbial communities
antagonistic to pathogens (Alvarez et al., 1995).
Peat moss-shrimp waste composts are commercially available. Such composts were shown to be of
high quality on the basis of criteria accepted by horticultural industries (Mathur et al., 1988). These could
be used as efficient fertilizers (Houtin et al., 1995).
However, water extracts made from a few commercial brands of shrimp waste-based composts exhibited
no inhibitory effect on the growth of oomycete plant
pathogens (Labrie, 1998). In such composts, no oligomeric forms of chitin could be detected suggesting
that the latter compound is completely degraded when
compost maturity is reached (Roy et al., 1997). Since
chitin derivatives can both inhibit the growth of several
plant pathogenic fungi and induce plant defense mechanisms (Kendra and Hadwiger, 1984), their presence
in mature composts could be an asset. Roy et al. (1997)
developed a two-phase composting procedure using
shrimp shells as an amendment to partly composted
biomass. The method consists of mixing peat moss,
sawdust and a nitrogen-rich biomass such as chitinous waste or cow manure and composting them until
the first thermophilic phase is terminated. The immature compost is then amended with shrimp waste,
thus inducing a second thermophilic phase. Roy et al.
(1997) showed that the chemical and physical properties of these biphasic composts were in agreement
with the criteria for the AA compost class (Bureau de
normalisation du Qubec, 1996). The purpose of this
work is to analyze the biological properties of biphasic
composts and to determine their suppressive capacity
against oomycete plant pathogens.

Materials and methods

Two phase composting
SP2 W2 +S compost was produced by mixing shrimp
waste, brown peat moss and pine sawdust in a 1:2:2
volumetric ratio as described by Roy et al. (1997).
After the thermophilic phase (57th day), maturing
composts were amended with one-third volume of
shrimp waste. Laboratory-scale composting was done
in adapted 4-liter Nalgene jars as previously described
by Roy et al. (1997). The jars, filled with biomass,

were placed in an incubator to impose a temperature pattern consisting of mesophilic phases (30 C)
separated by two thermophilic phases (50 C). Temperature was first set at 30 C for 14 days and was
then progressively raised to reach 50 C at the 20th
day of composting. The first thermophilic phase lasted 22 days after which the temperature progressively
decreased to 30 C. This temperature was maintained
between the 49th and 56th day of the composting process. After this second mesophilic phase, temperature
progressively increased to reach 50 C at the 64th
day. The second thermophilic phase lasted 14 days.
Finally, the temperature decreased to 30 C from the
79th to the 84th day and was maintained at 30 C
for 6 additional days. The jars were connected to a
forced aeration system (Roy et al., 1997). Throughout
the composting process, moisture content was measured and maintained around 50% to avoid drying.
Laboratory-scale composting was carried out to follow
microbial populations and chitin degradation since it
was previously shown that composts produced at both
laboratory and 20 m3 scales exhibited similar physical and chemical properties (Roy et al., 1997). To
analyze the effect of composts on plant diseases, largescale composting was carried out as described by Roy
et al. (1997). Briefly, 20 m3 piles containing either
shrimp waste, brown peat moss and pine sawdust in
a 1:2:2 volumetric ratio (SP2 W2 +S compost) or cow
manure, brown peat moss and pine sawdust in a 1:1:1
volumetric ratio (MPW+S compost) were established
outdoor on a concrete platform and protected from rain
by a roof. After the first thermophilic phase (at day
50), the partly composted biomass was amended with
shrimp shell-based waste in a proportion of 30% of the
initial dry weight of the composted primary biomass.
Oxygen content was monitored and maintained above
10% with a vacuum-induced forced aeration system.
Mechanical turning of the compost pile (at days 14,
30, 50, 62 and 74) also provided aeration. Moisture
content was measured every 5 days and maintained
between 45% and 60% by addition of water, to prevent
desiccation.
Determination of cellulolytic, chitinolytic and total
microbial numbers
Counts of cellulolytic, chitinolytic and total microbial populations were estimated throughout the composting process using serial dilutions and enumeration on cellulose agar, chitin agar and GPCA medium, respectively, as previously described by Roy

29
et al. (1997). Plates were incubated at 30 C (mesophilic micro-organisms) or 50 C (thermophilic microorganisms) for 25 days. Microbial counts were expressed as cfu per g of dry mass of compost.
Fatty acid analysis of compost microbial populations
Microbial cells present in compost samples (SP2 W2
and SP2 W2 +S) were separated from compost particles
using the protocol of Steffan and Atlas (1988) modified as follows. Compost samples (10 g) in 50 ml of 0.1
M sodium phosphate buffer (pH 5.7) were homogenized in a blender at low speed for 1 min. This step was
repeated three times and the mixture was cooled on ice
between each treatment. SDS was added at a final concentration of 0.8% (w/vol) and the mixture was mixed
in the blender for 5 s. The homogenized suspension
was then centrifuged 30 s at 500 rpm. The supernatant
was centrifuged again at 10 C for 30 min at 10 000
g. The cell pellet was resuspended and washed twice
in sodium phosphate buffer. Fatty acids were extracted
from the microbial cells according to Bligh and Dyer
(1959) and derivatized to their methyl esters according
to the protocol of Miller (1982). Main microbial fatty
acid methyl esters [iso 15:0, anteiso 15:0, 15:0, iso
16:0, 16:1, iso 17:0, anteiso 17:0, 17:0, 17:0 cyc, 18:0,
18:1, 18:2] (Pennanen et al., 1996) were identified on
a Hewlett-Packard HP 6890 series gas chromatograph
system equipped with a 30 m 0.25 mm silica SPBTM
column (internal diameter of 0.25 m) by comparing
their retention times to those of standards (Metraya,
PA, USA). Chromatography runs lasted 45 min with a
helium flow of 20 cm/s. Temperature was set at 4 C
for 4 min and increased at 4 C/min until reaching 250
C. Each experiment was carried out in duplicate.
Estimation of oligomeric chitin levels in composts
The content of the oligomeric form of chitin was estimated by measuring the -N-acetylglucosaminidasehydrolysable fraction (NHF) of compost samples
using the enzymatic technique described by Roy
et al. (1997). This method is based on the principle that chitin oligomers are soluble in water and,
therefore, can be hydrolyzed by the enzyme N-acetylglucosaminidase to generate N-acetyl--Dglucosamine monomers. The amount of monomers
liberated by such an enzymatic treatment from a
compost sample provides an estimation of the content of chitin in the oligomeric form. Briefly, 0.5
g samples of ground and dry compost were suspended in acetate buffer pH 5.5, vortexed to solubilize

the chitin oligomers and treated for 1 h at 50 C

with 0.1 unit of purified -N-acetylglucosaminidase
from Streptomyces lividans (Roy et al., 1997). After
centrifugation, the concentration of N-acetyl--Dglucosamine in the supernatant was determined using
the colorimetric method of Reissig et al. (1955). This
method took into account the naturally present N-acetylglucosaminidase by subtracting the average
optical density of the control tubes from that of the
test tubes. The NHF was expressed in g of N-acetyl-D-glucosamine liberated by g of dry compost.
Growth of oomycete plant pathogens in the presence
of compost extracts
Water extracts were prepared by adding 2 g of dry
compost to 25 ml of sterile water. Compost suspensions were incubated under darkness for 15 h with
shaking (300 rpm) at room temperature. They were
then centrifuged for 10 min at 3900 rpm. Supernatants
were sterilized by two serial filtrations, first on 0.45
m and then on 0.22 m pore diameter filters. These
compost extracts were tested for their effects on the
growth of two oomycete plant pathogens, Phytophthora fragariae var. rubi strain 390 (Valois et al.,
1996) and Pythium ultimum strain 447 (Laboratoire de
Diagnostic, MAPAQ, Sainte-Foy, Canada). Compost
extracts were added to potato dextrose broth (PDB)
(Difco, Detroit, USA) at 15% (vol/vol) final content.
Water was substituted for compost extracts in control
treatments. PDB mixtures (15 ml) were placed in 10
sterile petri dishes which were then inoculated with
a 6-mm-diameter agar plug excised from the edge of
a 2-week-old potato dextrose agar (PDA) (Difco, Detroit, USA) plate culture of Phytophthora or Pythium.
After a 3-day incubation at 30 C, fungal mycelia
were harvested by filtering through Whatman number
1 paper (Whatman International Ltd., Maidstone, England) and dried overnight at 60 C and weighed. Five
replicates by batch of compost were carried out. The
experiment was repeated twice.
Effect of compost-amended potting medium on
Pythium damping-off
The ability of compost-amended potting medium to
suppress Pythium damping-off of cucumber was analyzed. Mature SP2 W2 +S and MPW+S composts were
compared to a commercial brand of compost made
from shrimp waste and peat moss. The plant growth
substrate was composed of sand, potting soil and
vermiculite in a volumetric proportion of 2:2:1 and

30
subsequently amended with 25% (w/w) test compost,
on dry weight-basis. Plant growth substrate (250 ml)
was added to 10 cm-diameter pots and then saturated with water. Ten cucumber seeds cv. Sweet Slice
were deposited on the plant growth substrate and then
covered with an additional 50 ml of plant growth substrate. The pots were covered with transparent lids.
Cucumber seeds were allowed to germinate 5 days in
a growth chamber kept at 25 C with 90% humidity
and a photoperiod of 16 h. The plant pathogenic Pythium ultimum 447 was then added to the seedlings.
The Pythium inoculum was prepared by adding a 6
mm-diameter agar plug taken from the edge of a 2week-old plate PDA culture to 60 ml of plant growth
substrate supplemented with 6 g of oat flakes. Inoculum was deposited immediately after preparation
at the surface of each pot. Control treatments were
done in the same way but a sterile agar plug was used.
Inoculated and control pots were incubated for 1 day
at 15 C and 6 additional days at 21 C during the
light period and 18 C during the dark period. Pots
were arranged as a completely randomized block with
six replicates. The proportion of dead seedlings was
recorded after 4 days. A 2 test was carried out to
compare the different treatments. This experiment was
carried out twice.
Results
Oligomeric chitin levels and oomycete growth
inhibition property of compost samples
The SP2 W2 +Scompost was compared to a commercial brand of compost made from shrimp waste and
peat moss for the effect of its water soluble elements on the growth of Phytophthora fragariae var.
rubi and of Pythium ultimum. Water extracts from
the SP2 W2 +S compost (85th day) were associated
with 16%1.10 and 9%1.07 growth inhibition of
Phytophthora fragariae var. rubi and of Pythium ultimum, respectively (Table 1). In contrast, the commercial compost had a positive effect on the growth of
both pathogens (Table 1). It was previously reported
that the commercial compost did not contain oligomeric form of chitin (Roy et al., 1996). Similarly,
no oligomer of chitin was detected in the SP2 W2 +S
compost at the end of the composting process. However, oligomers of chitin was detected during the
composting process (Figure 1).
Detection of oligomeric chitin began during the
first thermophilic phase. The NHF value of samples

taken at the 25th day of composting was of 1 g Nacetylglucosamine (NAG)/g of dry compost. A NHF
peak (9 g NAG/g compost) was reached on the 32nd
day. The NHF value decreased to 2 g NAG/g compost between the 32nd and 40th day of composting
and then a slow but constant decrease was observed.
NHF value reached a second peak 17 days after the
shrimp waste amendment occurred on the 57th day
(Figure 1). The samples of the SP2 W2 +S compost
taken out during the composting process also served
to estimate the effect of their water soluble elements
on the growth of Phytophthora fragariae var. rubi.
Phytophthora growth inhibition by compost extracts
follows a pattern similar to that of the NHF. Compost
extracts exhibited highest growth inhibition capacity
when NHF values reached peaks. However, NHF
values reached maximal values during the first thermophilic phase while growth inhibition was higher during
the second thermophilic phase (Figure 1).
Micro-organisms inhabiting composts
Total microbial counts of the SP2 W2 +S compost were
5 1011 cfu/g at the beginning of the composting
process. Chitinolytic and cellulolytic organisms each
represented some 2% of the initial population. Microbial counts decreased during composting to 1
109 cfu/g after 57 days (Figure 2A). Counts of cellulolytic and chitinolytic micro-organisms were similar
throughout the first composting phase. Counts for both
groups were 1 1010 cfu/g before the first thermophilic phase and subsequently stabilized at 1 109
cfu/g (Figure 2A). The shrimp waste amendment was
shown to cause an increase of total, chitinolytic and
cellulolytic populations. However, the increase of cellulolytic populations was not as important as that of
total or chitinolytic populations that increased 100 fold
during the 5 days following the amendment. Then,
chitinolytic population decreased rapidly while total
microbial counts stabilized at about 1 1010 cfu/g
(Figure 2B).
Fatty acid analysis was also carried out to evaluate the variations in microbial populations during
composting of shrimp waste, peat moss and sawdust.
The proportion of each of the 11 fatty acids analyzed did not significantly vary between the 8th and
the 56th day of composting; even the passage to the
thermophilic phase did not cause a noticeable perturbation of the fatty acid profile. However, a shrimp
waste amendment to the partially composted biomass
did modify the microbial fatty acid profile. After the

31
Table 1. Growth of Pythium ultimum 447 and Phytophthora fragariae var. rubi in the presence of compost extracts
Compost extract a
Pythium ultimum
Control b
Commercial compost
SP2 W2 +S

68.1 b c
102.2 c
61.9 a

Weight (mg)
Phytophthora fragariae var. rubi
60.3 b
74.8 c
50.7 a

a Compost extracts were prepared by immersing 2 g of dry compost to 25 ml of sterile water for 15 h with shaking. They were filter-sterilized
prior their addition to potato dextrose broth at 15% (vol/vol) final content.
b Water was substituted for compost extracts in control treatments.
b Data are the mean of five replications. Numbers of a column accompanied by a same letter did not significantly differ (t test, p < 0.05).

Figure 1. Comparison between the NHF values of water extracts from SP2 W2 +S samples taken throughout the composting process and the
growth inhibition of Phytophthora fragariae var.rubi caused by these extracts. The arrow indicates the time of the amendment. The data
presented are the mean of two independent experiments. In each experiment, five replications of the inhibition assay were carried out while
NHF assay was done in triplicate.
Table 2. Table 2. Effect of composts on damping-off of cucumber
caused by Pythium ultimum 447
Compost type

Mortality rate (%)

Experiment 1
Experiment 2

Commercial compost
MPW+S
SP2 W2 +S

96.9 a a
83.6 b
62.9 c

73.5 a a
51.9 b
39.3 c

a Numbers accompanied by a same letter did not significantly

differ ( 2 test, p < 0.05).

amendment, branched fatty acids represented a mean

of 32% of the total amount, up from a mean of 17%
before the amendment. The proportion of branched
fatty acids remained stable (16%) when the shrimp
shell amendment was omitted (Figure 3).

Effect of composts on damping-off of cucumber

The effect of three composts (commercial compost,
SP2 W2 +S compost and MPW+S compost) on the survival of cucumber seedlings sown in the presence of
Pythium ultimum was evaluated in two separate experiments. The mortality rate of cucumbers raised in
non-infected substrates containing any of the three
compost was very low and varied between 0.0 and
1.2%. When the pathogen was added to the growth
substrates, the mortality rate increased considerably.
The mortality rate was highest in substrate containing
commercial compost (Table 2); 97% and 74% mortality was recorded for the first and second experiments,
respectively. Damping-off was less severe in the presence of SP2 W2 +S and MPW+S composts. SP2 W2 +S
was the most effective compost to reduce infection
caused by Pythium (Table 2). Observed mortality rates

32

Figure 3. Effect of a shrimp shell amendment on microbial population as determined by fatty acid analysis of compost micro-organisms. Primary biomass consisted of shrimp shells, peat
moss and sawdust in a 1:2:2 volumetric ratio. Primary biomass was
amended with one-third volume of shrimp shells at the 57th day of
the composting process. The arrow indicates the time of the amendment. The data presented are the mean of two experiments. Each
experiment consisted of two replications.

Figure 2. Enumeration of chitinolytic, cellulolytic and total micro-organisms throughout the composting of biomass consisting of
shrimp shells, peat moss and sawdust in a 1:2:2 volumetric ratio.
(A) Primary biomass was not amended; (B) Primary biomass was
amended with one-third volume of shrimp shells at the 57th day of
the composting process. The data presented are the mean of two
replications.

were 63 and 39%, for the first and second experiment,

respectively.

Discussion
Two independent experiments were conducted to analyze the effect of chitin waste-based composts on cucumber damping-off. In both experiments, SP2 W2 +S
and MPW+S composts reduced the incidence of
damping-off when compared to a commercial brand
of compost made from shrimp shells and peat moss
(Table 2). However, mortality rate of cucumber seedlings was lower during the second experiment. The
fact that each experiment was carried out in different
laboratories (Dpartement de biologie, Universit de
Sherbrooke and Institut de Recherche et de Dvelop-

pement en Agroenvironnement, Sainte-Foy) might explain the discrepancy between the two experiments.
Different lots of seeds as well as of culture media
have been used in each laboratory. Moreover, environmental conditions encountered by the plants and by
the pathogen might have slightly differed in the two
locations.
It was previously reported that some commercial
brands of compost contain no oligomeric forms of
chitin (Roy et al., 1997). The presence of oligomeric
chitin in a compost could confer it with suppressive properties since oligomer derivatives are known
to protect plants against pathogens by inhibiting the
growth of fungi and/or by inducing the plant defense
mechanisms (Kendra and Hadwiger, 1984). Nevertheless, the lower incidence of damping-off of cucumbers
in the presence of SP2 W2 +S is unlikely due to the
presence of oligomeric chitin in the composts. Even
though oligomeric chitin was detected during the two
thermophilic phases of the composting process, NHF
decreased progressively during the consecutive mesophilic phases to become nil. During the first phase of
the composting, the level of growth inhibition exerted
by compost water extracts followed a pattern similar to
that of the NHF suggesting that the presence of oligomeric chitin was responsible for Phytophthora growth
inhibition. However, inhibition caused by samples
of the second phase of composting could not only
be attributed to the presence of oligomeric chitin in
water extracts, since the inhibitory effect recorded

33
during this second phase did not increase proportionally to NHF values. Furthermore, NHF was nil at
the end of the second phase while the water extract
still inhibited Phytophthora and Pythium growth. The
living components of SP2 W2 +S compost are not directly responsible for this growth inhibition since the
extracts were filter-sterilized. Rather, chemical compounds present in compost would have affected the
development of Phytophthora and of Pythium as previously suggested by Cronin et al. (1996) for other plant
pathogens.
The suppressive property of these composts might
result from the two-phase composting process. This
study provides evidence that shows the microbial
properties of compost were modified when shrimp
waste was added to partly composted biomass. First,
counts of microbes in an amended compost were
higher than in a non-amended compost using the
same primary biomass. Second, the proportion of
branched microbial fatty acids increased following
the amendment. Iso and anteiso fatty acids are characteristic of Gram-positive bacteria (Suzuki, 1988).
The shrimp shell amendment thus appears to promote
the proliferation of Gram-positive bacteria, a group
of bacteria producing more than 70% of known antibiotics (Strohl, 1997). Several reports established
that numerous Gram-positive bacteria could act as
biological tools against oomycetes (Crawford et al.,
1993; Emmert and Hadelsman, 1999; Hadelsman et
al., 1990; Sutherland and Papavizas, 1991; Toussaint et al., 1997; Valois et al., 1996) and production of antibiotics is an important asset for most
of these biocontrol agents (Emmert and Hadelsman,
1999; Toussaint et al., 1997) Antibiotics as well as
antibiotic-producing Gram-positive bacteria such as
Bacillus and actinomycetes have been previously isolated from composts (Craft and Nelson, 1996; Fiddaman and Rossall, 1994). The fact that water extracts
of SP2 W2 +S samples taken during the second composting phase exhibited a higher level of Phytophthora
growth inhibition than the ones from the first phase
might result from a higher concentration of antibiotics or other toxic molecules produced by proliferating
Gram-positive bacteria.
MPW+S, a compost for which chitinous matter
was omitted from the first composting phase, also controls damping-off of cucumber, although to a lesser
extent than SP2 W2 +S. This suggests that chitinous
compounds are not essential in the first composting
phase to develop a suppressive compost. It is not yet
known if any type of composted primary biomass,

once amended with chitinous material to induce a

second thermophilic phase, will produce a compost
capable of suppressing diseases caused by oomycetes.
Except for shrimp waste or cow manure, the primary
biomass of SP2 W2 +S and MPW+S composts were
identical. Primary biomass of both composts are rich
in lignocellulosic matter. Composts prepared from material with high cellulose content such as sawdust are
not usually suppressive to plant disease (Hoitink et
al., 1987). This work suggests that a two phase composting process consisting in a chitin amendment to
partly composted lignocellulosic biomass might confer suppression properties to the resulting compost.
The suppression mechanisms of two-phase composts
have to be further elucidated, but apparently involve
the proliferation of Gram-positive bacteria within the
microbial populations of compost. The occurrence of
proliferation of Gram-positive bacteria simultaneoulsy
with the growth inhibition property of compost extracts suggests to us that suppression might be due
to the production of compounds restricting oomycete
proliferation by Gram-positive bacteria.

Acknowledgements
The authors gratefully acknowledge the financial support of the Conseil des Recherches en Pches et en
Agro-alimentaire du Qubec (CORPAQ). We thank A.
Gauthier for critically reading the manuscript and L.
Ct for his assistance in compost preparation.

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