United States
Environmental Protection
Agency
Water Engineering
Research Laboratory
Cincinnati, OH 45268
Research and Development
EPA/600/S2-88/024 August 1988
 Project  Summary

               of
 Sludge  on  Soilborne  Plant
 Pathogens

 H,A. J, Hoitink and A, F. Schmitthenner
   Efficacy of composted municipal
sludge (CMS) for suppression of
Phytophthora root rot of soybean In
field  soil and for suppression of
Rhizoctonta  and Pythium diseases of
ornamental plants  produced In
container        was  investigated
over a 3-year period.
   CMS increased yields of soybean
by Improving soil fertility and/or by
partial control of Phytophthora root
rot {PRR).  Disease-enhancing
effects of salt  in the CMS  were
controlled with a selective fungicide.
In the  absence of the fungicide,
beneficial effects were obtained only
by application of CMS at  least  3
months prior  to  planting  with
cultivars tolerant to PRR, Increased
yields of soybeans were observed
when high rates of CMS were applied
to corn  crops and soybeans were
grown in the next year without further
CMS application. No residual effects
were observed from high rates of
CMS applied to soybeans.
   All container media prepared
with CMS cured 4 months or more
and stored 4 weeks after formulation
became consistently suppressive to
Rhizoctonia and Pythium diseases of
ornamentals at  levels  adequate to
avoid losses under   commercial
conditions.  An unknown  beneficial
microflora involved In suppression, if
present, survived in the outer low-
temperature layer of curing  piles
only.
   Several bacterial  and fungal
isolates were identified that ef-
fectively  induced suppression to
 Rhizoctonia and  Pythium "damping-
 off" in CMS  media.  The relative
 contributions  of  these microbes to
 the overall  suppressive effects
 remain to be determined.  Most
 bacterial  strains  were more ef-
 ficacious  in  combination  with  an
 isolate of  Trichoderma. A synergistic
 interaction was found  among  an
 isolate of  Trichoderma ham a turn and
 of  flavobacterium  balustinum for
 suppression  of  Rhizoctonia  damp-
 ing-off. These isolates, if added to
 conducive CMS media, consistently
 rendered  them suppressive  in the
 absence of the  4-week incubation
 period required for  development of
 natural suppression.
    This  Project  Summary was
 developed by EPA's Water Engineering
 Research  Laboratory,  Cincinnati, OH,
 to  announce key  findings of the
 research  project that is fully doc-
 umented in a separate report of the
 same title (see Project Report order-
 ing Information at back}.
 Introduction
    A significant number of munici-
 palities in the United States are using the
 composting process for further treatment
 of municipal sewage sludge. Composted
 municipal sludge (CMS) has been utilized
 effectively as a source of piant nutrients,
 and the organic matter in CMS may
 improve physical  and biological
 properties of soils.
    Severity  of a number  of plant
 diseases can be affected by CMS. Some

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are decreased in  severity,  whereas
others are increased as a result of field
soil amendments. The mechanism of this
variability is not understood. However,
suppression  of  disease  has  been
associated  with  increased microbial
activity of treated soil. Severa! micro-
organisms  have  been  isolated from
composts  that are  involved   in  the
suppression of soilborne  diseases in
soils or  container  media amended with
composts. Their fate during composting
of municipal sludges is unknown.
Phytophthora Hoot Rot of
Soybeans
    Phytophthora root rot of soybean is
one of the most severe diseases of this
crop in  the  northern soybean  growing
region. Approximately  10 million  acres
are  infected  and  can  be severely
damaged in wet years. Presently, control
of root rot is  based on  resistance of
cultivars which has  been overcome  by
new  Phytophthora races; application of
the  fungicide  metalaxyt  (a potential
ground-water pollutant) to soil;  or  a
combination of good drainage, complete
soil tillage, rotation, and metalaxyl  seed
treatment  of  highly  tolerant {least
susceptible)  cuitivars. The latter  inte-
grated control package is not available to
farmers who wish to reduce soil erosion
through conservation tillage,
    Additional durable and nonpolluting
options are needed for control of this
important  disease.  Phytophthora
diseases  can be controlled by adding
certain organic amendments to  the sotl.
One  way to rapidly increase soil organic
matter is  with  manures  and  CMS.
Manure is not readity available to farmers
that  focus on  cash cropping.  On the
other hand, CMS, derived from municipal
sludge, is available in many areas of the
United  States where  soybeans are
produced, CMS can be spread  on  fields
with  a conventional manure spreader,
thus  making  widespread  utilization
possible.  However,  CMS  may have  a
high  salt  content,  which can create
adverse  effects.  Both  high  fertilizer
application  rates  and high  chloride
concentrations increase PRR of soybean.
    The experiments reported here were
designed to investigate three areas. First,
effects of various CMS rates  on root rot
severity  and  yield  of  soybean in
Phytophthora-infested  soil were exam-
ined. Second, the  effect of  application
timing was  determined.  Finally, the
residua! effects of  CMS applied  during
the previous season were evaluated.
Suppression of Rhizoctonia and
Pythium D/seases of
Ornamenfa/s Produced in
Container Media
    Composts prepared from a variety of
tree  barks are utilized  successfully  for
suppression of soilborne diseases of
ornamental plants  produced in container
media. These composts are particularly
effective  for control  of  Rhizoctonia and
Pythium  diseases when  microbial an-
tagonists  have recolonized the  compost
after peak heating has  occurred.  The
mechanisms  of  suppression  involved
vary with the pathogen,  the source or
type of compost used,  and the  level of
decomposition achieved  during  the
composting process. This portion of the
study  shows  that  container  media
amended  with CMS  can  be  used  ef-
fectively  for  suppression  of Thizoctonia
and Prrthjum diseases of greenhouse and
nursery crops if certain precautions are
taken  to overcome the detrimental
effects imposed on microbial antagonists
by  the  long-term  high  temperature
treatment (>55°C) necessary to ensure
fecal pathogen and parasite destruction.
In addition,  beneficial  microorganisms
involved  in the suppression of  Rhizoc-
tonia and  Pythium  diseases  were
identified,
Objectives
    The objectives of research reported
here were: 1} to determine the effects of
CMS on  soybean  PRR  severity  in  the
field;  2) to develop procedures for  the
formulation of container media with CMS
that consistently  suppress  diseases
caused by soilborne plant pathogens of
ornamental plants;  and 3) to  determine
the nature  of  the  beneficial microflora
involved  in suppression  of  these  dis-
eases,


Methods

Cftaracferisf/cs of the Compost
    Composted municipal  sludge  was
obtained  from the Columbus Southerly
Composting  Facility where the  aerated
static pile composting process has been
used on  a daily basis since 1980.  Raw
sludge cake, consisting of a 1:1 ratio (dry
wt  basis)  of primary sludge and waste
activated sludge,  was  dewatered  with
centrifuges  aided  by  polymer  as a
coagulant. Sludge cake,  averaging  16%
to  17%  total  solids,  was mixed  with
hardwood chips (1 in. to  2 in. chips) at a
ratio of 5 volumes of chips to 1 volume of
sludge.  Mixing  was accomplished  with
front-end loaders  in a covered  mixing
area. The composting  process generally
was complete after 21 days. Piles were
then processed and  stacked with  or
without  aeration  for  a  60-day  curing
period. Thereafter, the  mixture was dried
by passive solar  drying on  a concrete
pad.  Wood  chips were  recovered  by
screening  with  a  3'8-in. rotary  screen
Screened  compost was  then stored in
bays  in  a covered  shed-  A  weekly
composited  compost  analysis  was
associated  with each  bay of compost.
Compost  thus  prepared  was  used
throughout this work  A summary of the
chemical  properties  of  the  CMS is
presented in Table 1.


Soybean Phytophthora  Root
Rot
    Field experiments were  conducted
for 3  years at the Northwest Branch of
Ohio  Agricultural  Research  and
Development  Center  (OARDC)  of The
Ohio State  University  located at  Custar,
OH, in a Hoytville clay loam containing
50% clay, 40% si it, and 10% sand  The
plot area used  during the first  2 years
was  tiled, but gates  located at the tile
exits were closed to reduce drainage In
the third  year, the location was changed
so that the plots could be irrigated 3 days
after  planting  to  provide optimum
conditions  (or  Phytophthora infection
This  field was tiled, but the tiles were old
and nonfunctional
    Plots consisted of  8 rows, each 26 m
long and spaced 76 mm. Each plot  was
split  into two  treatments  of  four rows
each. In the first year the split treatments
were the cultivars Sloan (some tolerance
to PRR)  and OX 20-1 (no tolerance to
PRR). In the second and third years the
split  treatments consisted of the  cultivar
Sloan,  with  and   without  280  g
metalaxyl/ha (5.6 kg/ha of Ridomil  5G").
This systemic  soil fungicide  provides
good control of  Phytophthora  when
applied in  the  seed  furrow  at  planting
time.
    CMS application  rates were 0,  10,
20, arid  40 tonne,ha. Application times
were  preplant,  applied  within  1 week
before  planting to maximize  salinity
effects; and  early,  applied  at  least 3
months  before planting,   either  in
February  or  November,  to minimize
salinity. To determine  the fertility effects
        of trade names or cornrnercidl products
  does not constitute endorsement of recom-
  mendation (of use

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                       Table !      Characteristics of Municipal Sludge and
                                   from the Columbus Southwesterly Compost
                                                       Sludge"
                                 Composted Municipal Sludge
                                 Facility
                                      Composted Sludge

Volatile Solids %
Conductivity (mmhoS'Cmff
Cl (ng-mi
pH
W (°.'o, total Kjetdahl)
P (°A)t
K (%)
Ca (%}
Mg (%>
Cd (fflQtkrj)
Cr fmg.kg)
Cu (fflQ-kg)
Pb (mg-kg)
Ni (mg.kg)
Zn fmgftgj
Na Cmg'ht?;
Mean
71


5.8
6.5
3,2
? 5


8.8
131
218
170
45
1,057

flange
57-79


5.4-6.3
3.2-8,4
1, 6-5.8
0.2-2.2


2,7-26
72-290
1 30-270
92-560
18-74
550-1,900

Mean
59
10,5
370
7.5
4.5
3.3
1.6
3.6
0.86
20
262
255
274
65
1,664
776
Range
43-74
7.5-15,0
245-360
6.2-8.4
3.2-5.3
2.5-4.7
0.9-1.9
3.4-3.8
O.S4-0.8?
73-24
750-325
1 90-640
760-320
54-111
1.000-1,971
699-827
                       "The sludge was a 1:1 mixture of raw and waste activated sludges. Means are based on
                        64 samples. Analyses performed by City of Columbus Central Surveillance Laboratory.
                       tDetermined in a saturated pasts extract.
                       tAtt metal concentrations were determined after nitric perchloric ecid digestion.
of  CMS, plot soil was brought  into the
laboratory, mixed with the equivalent of
0,  10, 20, and 40 tonne dry wt CMS/ha,
planted  with soybeans under  conditions
that  promote  maximum Phytophthora
tosses, and,  after 2  weeks,  submitted to
the  Research   Extension  Analytical
Laboratory (REAL), QSU,  for  fertility
analysis {Table 2). Based on  this
analysis,  a  fertility  control preptant
treatment was   applied  consisting  of
superphosphate  (200  kg PgOs/ha) and
NaCI (327 kg/ha) which  was the  soluble
salt equivalent of 20  tonne/ha CMS.
    Disease analysis consisted of counts
of  healthy and diseased plants 5 weeks
after  planting   in  late October,  the
weights and  percentage moisture of the
seed from the two center rows  of  each
pfot were taken  to determine effects of

                          Tatjie 2,
    treatments on yield. Ail treatments were
    replicated  four  times.  Data  were
    subjected to an  analysis of variance with
    application times  and CMS rates as
    whole plots  and cultivars  of metalaxyl
    soil fungicide treatment as split pfots.


    Preparation of Container Media
        A CMS  container  medium  was
    prepared by mixing CMS with Canadian
    sphagnum peat  and perlite (1;2:1,v/v, pH
    5.5), This medium was not amended with
    fertilizer  since sludge compost media
    release  adequate  amounts of  plant
    nutrients for at least 6  weeks  after
    potting.  A peat container  medium was
    prepared from Canadian  sphagnum peat
    and perlite (1:1, v/v) and adjusted  to pH
    5.5 with  a 2;1 (w/w) mixture of cfolomitic
and  hydrated  lime. It was fertilized to
adjust fertility to levels identical to that in
the CMS medium. The peat medium is
conducive to Rhizoctonia  and Phthiurn
dam ping-off  and,  therefore, served  as  a
control in this work.
    Suppression  of Pythium  and
Rhizoctonia  damptng-off was  deter-
mined  with  cucumber  and radish
bioassays,  respectively. Inoeuta of
Rhizoctonia  so/an/  and  of  Pytftium
ultimum were added at rates  of 0.5 g/L
container medium.  This amount of
inoculum kilted 50% of  the plants  in  a
suppresstve  medium and all plants  in  a
totally  conducive  peat medium. Mean
disease severity ratings were determined
for various sludge  compost  and  peat
media  to compare efficacy of CMS for
suppression of these diseases.
Fertility Factors in HoyM/te Stity Clay Affected by Applications of
Composted Municipal Sludge
Rate
(dry tonngfha)
0
10
20
40
LSD (0.05)

P
134
214
353
575
93

K
498
501
545
642
55
kg/ha
Mn
19
20
28
39
8

Zn
96
105
139
222
58

8
1.8
2.1
2.4
3.1
0,26
WO3
340
370
532
S78
120

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                                Tafile 3.     Effect of Composted Municipal  Sludge  on Yield
                                           (kgltia) of Soybeans Under MM Phytophlhora Root
                                           Rot Pressure in 1984
                                                                     Variety
                                    Rate
                                (dry tonn&iba)
     Application Time
Sloan
(susc)
OX 20-8
  (very
  suscj
Means
                                                                   Yield (kg/ha)
                                     0

                                     ?0
                                                  F0584
                        2651

                        2590
          7959   230S

          1953   2272
20
40
0
10
20
40
Salt Control"
LSD =0.05
Feb84
Feb84
May 84
May 84
May 84
May 84
May 84

2849
3345
2779
2657
2923
308?
2583
499
1853
215?
2235
2076
1581
IS58
1612

2356
2751
2507
2366
2252
2322
2087
256
                                '200 kg of P^Os and 327 kg NaCtha, the soluble salt equivalent of
                                20 tortna CMS-ha
Interactions Among Beneficial
Microorganisms in Container
Media
    Bacteria and fungi were  isolated by
baiting with  radish and cucumber roots
produced  in  CMS  media  or with
Rhizoctonia and Pythium soit propagules
incubated  in  sandwiches  in  the
suppressive media. Pure cultures of 650
bacterial  isolates and 30  fungal isolates
were  then tested for their  abilities  to
induce suppression to  Rhizoctonia and
Pythium damping-off  using  the
bioassays described above. Spontane-
ous  rifampicin-resistant  mutants were
isolated from  the most effective bacterial
strains. The  root  colonizing  potential of
these mutants was  then  tested.  Finally.
various mixtures of bacterial antagonists
with  strain 382 of Trichoderma hamatum
were examined to reveal  synergistic
interactions among  antagonists  in CMS
media.
Results

Soybean PRR
    In 1984, the 40 tonne dry wt CMS/ha
treatment  applied  in  early  February
significantly increased  the  yield of the
moderately susceptible  cultivar  Sloan
(Table  3),  Yield  differences were not
found in  the  highly susceptible cultivar,
OX 20-8. There  were no differences in
stand or  plant death among any of the
variety-treatment  combinations.  The
lack of yield response  in  the highly
susceptible OX 20-8 could be explained
by  greater  hidden  root  rot  damage
offsetting a potential fertility response.
    High rates of CMS applied  preplant
(May)  did  not increase yield  of  the
moderately  susceptible cultivar Sloan
and  also significantly reduced  the  yield
of highly susceptible  OX  20-8. These
results  were not accompanied by
significant differences in stand  or  plant
death  from   PRR.  The salt-phosphate
treatment significantly reduced the yields
of both  cultivars. This  agrees  with
previous  findings  for effects of  chloride
on  soybean PRR  severity. The salt
content  of  CMS  applied preplant
probably increased hidden root  rot,
which prevented a  yield  response in
Sloan and decreased  yield in OX  20-8.
The yield of  Sloan in  the  high  CMS
treatment was significantly  better  than
that in  the   salt-phosphate treatment,
indicating a  response to organic matter
even in  the presence of  high  salts.
Cultivar OX  20-8  did  not  respond to
organic  matter in the presence  of sait.
Thus, the response of cultivars to organic
matter may  have been associated with
their  level  of  susceptibility  to
Phytophthora.
    In conclusion,  the  1984  season
demonstrated that  the yield  response of
soybeans may depend on susceptibility
                  to PRR  and application time.  Possible
                  negative effects of high salt in CMS could
                  be eliminated by early  application. To
                  separate the fertility and disease seventy
                  effects, experiments for  1985 and  1986
                  were changed  to incorporate control of
                  Phytophthora  with Ridomil, a  specific
                  fungicide, Ridomil applied  in the furrow
                  with the seed  at  planting  time  controls
                  PRR  damage  in all  but the  most
                  susceptible  cultivars,
                      In 1985 there were no differences
                  between Rrdomil-treated and untreated
                  plots  The  salt-phosphate treatment had
                  no effect on yield, Thus, in 1985,  PRR
                  did not occur  Both the 20 tonne ha and
                  40 tonne ha rates  of CMS significantly
                  increased yield (Table 4), The beneficial
                  fertility  effects  of CMS were clearly
                  evident  in  this year  in  the  absence of
                  Phytophthora dam age.
                      Residual effects of CMS  applied in
                  1984 were evaluated in 1985. There were
                  no differences among any  of the  CMS
                  rates  in soybean-soybean  plots (Table
                  4), All yield levels were low.  In the  corn-
                  soybean sequence, the high rale of  CMS
                  (40  tonne'ha)  applied to corn  in  1984
                  significantly  increased  the  yield  of
                  soybean in 1985 Thus,  the  residual
                  effects  of  CMS  were  only evident  if
                  applied to corn, which is the logical time
                  to apply CMS  in a cropping system.
                      In 1986, only two rates of  CMS,  0
                  and 40 tonne  dry wt'ha,  were compared.
                  Ridomil, application time,  and  salt-
                  phosphate treatments also were included.

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                              Table 4.     Effect of Composted Municipal Sludge on Yield (kg/ha) of
                                         Soybean Under Phytophthora-Free Conditions in  19B5
                                                          Crop and Time of Application
Sludge
(tonne/ha)
0
10
20
40
Salt Control'
LSD(O.Q5} = 270 Kg
Feb 94
Soybean
3073
3086
3060
3154
-

May 84
Soybean
2893
2965
2979
3113
2979

May 84
Corn
3370
3423
3423
3558
3370

Nov 84
Soybean
3208
3235
3504
3693


May 85
Soybean
3208
3316
3693
3706
3383

                              '200 kg of P2O5 and 327 kg NaCUha, the soluble salt equivalent of 20
                              tonne CMSiha.
The crop was irrigated with tO cm water
over  a 2-day  period  approximately 3
days after planting when the seedling tap
roots  were  about 5 cm long  Flooding
induced conditions favoring PRR. Yield
levels were not high  in this experiment
because  of a  9-week drought between
July and  September and fertility benefits
were not  evident.  However, a  significant
disease  response to treatments  was
evident.
    Preemergence and postemergence
damping-off  occurred in  plots  not
treated with Ridomif  (Table  5), Stands
were poorest  in plots with  CMS applied
preplan! and  best  in plots treated with
CMS early, Ridomit controlled damping-
off and resulted  in  good stands and
improved yields in all sludge treatments.
In plots not treated with Ridomil, only the
early   CMS application  significantly
increased  yields compared  to  the
preplant  application  and  the  controls,
Under  this severe disease  pressure,
early  application  of  CMS  increased
yields  over the control  as  much as
treatment with Ridomil.  Response  from
CMS  and  Ridomil  was  not  additive,
possibly  because  the   late  season

                         Table 5.
   drought prevented a  fsrtiiity  response.
   The 1986 data confirm the importance of
   time of application of CMS.
       The following conclusions on PRR in
   soybeans can be drawn from these data.

     1. CMS  applied just  prior to  planting
       may increase Phytophthora damage.
       Yields  will  be  increased  or
       decreased depending upon  severity
       of Phytophthora.
     2. A salt-phosphate mixture equivalent
       to fertility  in CMS,  has  the same
       erfect as CMS applied immediately
       before planting,
     3. CMS appfied 3 to  6  months before
       planting reduced  Phytophthora  and
       increased yield.
     4. CMS  applied in  the  absence of
       Phytophthora increases yields  even
       in very susceptible cultivars.
     5 Response   to  40 tonne  dry  wt
       CMS ha is  better  than to 20 or 10
       tonne dry wt ha.
     6. Ridomil applied  at  recommended
       rates reverses the  negative effects of
       CMS  applied immediately  before
       planting.
Effect of Compost  Curing Time
on Suppressiveness of CMS
    All batches of CMS stored in curing
piles with  temperatures  in the center of
the pile higher than 60°C and  formulated
into a  CMS medium, were consistently
suppressive  to  Pythium  damping-off.
However,  these  CMS  media varied in
their effects  on  Rhizoctonia  damping-
off- CMS cured 4 months or more in piles
and  formulated into  media, became
consistently  suppressive  to Rhizoetonia
damping-off within an  additional  4
weeks of storage.
    The  environment in which  the
compost   was   cured   affected
Suppressiveness. CMS stored  next to
piles of bark  compost  suppressive to
Rhizoctonia  damping-off,  became  sup-
pressive  earlier than  CMS  stored in
isolated locations.  Microorganisms  iso-
lated from the  suppressive  CMS  that
could cause this  effect were Trichoderma
spp.  and  a number  of  bacterial
antagonists.  Without this recolonization of
CMS  with  antagonists  of  R. solani,
Suppressiveness did not develop.
Effect of Composted Municipal  Sludge on Stands (ptants.26 m of
Row  Spaced  76 cm) and Yields (kgtha) of Sloan Soybean Under
Severs Phytophthora Root Hot Pressure in 1986
                                                Stand (plants 26 m rows)
                                       Yield (kg ha)
Sludge
(tonne; ha)
0
40
0
40
Salt Control"
Time of
Application
Feb 86
Feb 86
May 86
May 86
May 86
No
Rid omit
211
226
176
105
207
Ridomil
352
318
346
312
354
Mean
281
272
261
209
280
No
Ridomil
984
2056
627
512
815
Ridomil
2420
2615
2251
2736
2440
Mean
1705
2332
1442
1624
1624
                          LSD (0.05)
               89
                                                                            607
                                                                                        337
                          "200 kg of P2&5 and 327 kg NaCl^ha, the soluble salt equivalent of 20 tonne
                          CMS'ha

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                     Table 6      Suppress we ness of a Nursery Sludge Compost and a Conducive Peat Container
                                 Medium to Pythium and Thizoctoma Damping-Off Over a 2-year Period
                                                           Disease Seventy Rating?
Medium
Sludge
Compost
Peat
Control
Treatment
Chech
Infested
Check
infested
May
7.7
1.8
1.2
3.9
July
1.0
2.6
1.1
4.0
Nov
1.0
23
17
3.5
LSD (P =0.05)
Oct
1
2.
1
3.
0.8
7
7
2
a

May
1.5
3.7
r.3
3.9

July
1
2
7.
3

2
6
3
9
0.3
Nov
12
23
1 7
36

Oct
1 r
29
! 1
38

                     "Infested with 0.5 g Rhizoctonia solan! or Pythium ulttmum soil inoculum per L container medium.
                     fOisease seventy ratings: 1 = symptomless; 2 ^diseased but not damped-off.
                      3 =postemergence damp/rrg-otf; 4 =pre emergence damping-off, based on five pots each
                      planted with 32 radish seed. May, July and November represent 1984. October represents
                      1985 values.
Suppression on Diseases During
the Production of Potted Crops

    Pythium  diseases  caused  by P.
ultimum or  P.  aphanidermatum in
floricultural pot crops were suppressed in
CMS  media  throughout a  5-month
production  period.  Similarly,  Pythium
root  rot caused  by P.  irregulare in
nursery  crops was suppressed over a
2-year  production  period  (Table  6),
             Diseases caused by R. solani were also
             suppressed in both floricultural (Figure 1}
             and nursery crops  (Table  6), but  only
             consistently  if the formulated container
             medium  was  stored  4  weeks  before
             utilization.

                 It was concluded that CMS becomes
             naturally  suppressive to Pythium damp-
             ing-off but that 4 months  of curing  is
             required  followed  by  an  additional  4-
week  storage  period  of  the  formulated
medium  required to ensure suppres-
siveness  to  Rhizoctonia dampmg-off

Efficacy of Microbial
Antagonists
    T.  hamatum  382  added   as an
antagonist by  ttself to a conducive  CMS
medium did  not induce a significant level
of suppression to Rhizoctoma dampmg-
off (Table 7). However, a combination of
                 Table 7.
Suppression of Rhizoctonia Damping-Off Induced by Bacterial Antagonists Alone and in
Combination with Tnchoderma hamatum in a Sludge Compost Container Medium
                                      	Disease Severity Patmgt	
                                 Bacterial Antagonist"
                 LSD 005
                                                                    Bacterial Antagonist
                                                                         Alone
                                                          Bacterial Antagonist
                                                           with T, hamatum
Wone
Bacillus cereus 106
Enterobacter cloacae 127
0. cloacae 3)3
Flavobacterium balustinum 299 (ATCC53198)
Janthinobactenum livid urn 275
Pseudomonas ftuorescens biovar til, A9i
P. (lucrescens biovar V, Ai
P. fluorescons biovar V, A498
P. putida 305
P. putida 375
P. putida 37J
P. stut2ert280
Xanthomonas maltophilia 76 (ATC53199)
2.9
3.0
2.6
21
3.1
2.7
2.9
2.4
2.7
22
23
23
24
30
26
20
2.6
2 J
2.1
25
2.4
26
23
1 3
2 1
25
27
2.4
                                                                                     04
                 "Bacterial antagonists were adcfed to the heated (60"C, 5 days) sludge compost container medium at
                   initial population levels of 10SCFU g dry wt T. hamatum was added af TO4 CFUg dry wt.
                 flnfested with 0.5 g Rriizoctonia solatu soil inoculum:L container medium. Mean disease seventy rating
                   determined 7 days after incubation at 26"C from Five pots planted with 32 radish seeds each.
                   1 -symptomless^ 2 -diseased but not damped-off: 3 -postemergence damping-off; and
                   4 -preemergence damping-off Mean disease severity in the container medium not infested with R.
                   solani was  1.2

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                                            8.O-
    Aug  Sept   Oct    Nov   Dec
               Sampling Date
Figure  1.    Suppresstveness of s peat and
            a sludge compost  medium
            {CMS} to Rhizoctonia damping-
            off during production  of poin-
            settta plants. Disease severity
            rating: 1 ~symptomtess; 2-dis-
            eased but not damped-ofl,
            3=post-emergence damping-
            off; and 4-^preemergence damp-
            ing-olf. Infested  with 0.5 g
            Rtzoctonia inoculum  per liter
            container medium. ISOoos for
            all  sampling  times  com-
            b/t-ied-0.4.

Flavobacterium  balustinum  strain  299
with  T.  hamatum  382  significantly
(P = 005) reduced damping-off, even
though each bacterial antagonist by ttseif
was not  effective.  This bacterial antag-
onist  significantly enhanced  population
development of the T.  hamatum strain
early  after its addition  to the compost-
amended medium  (Figure 2).  A syner-
gistic effect  was established, therefore.
This effect occurred in compost cured 4
months that was conducive if  not infested
with  the antagonists   Several  other
combinations of bacterial antagonists and
T. hamatum 382 were identified  that had
similar synergistic  interactions  in  sup-
pression of  Pythium damping-off in the
CMS medium.
    It  was  concluded that  microbial
antagonists  can  be utilized  to  develop
predictable   levels of  suppression to
Rhizoctonta  damping-off  in  container
media amended  with  conducive  CMS.
This process eliminates  the need  for
storage of formulated CMS media.
    Tho full report was submitted in
fulfillment of Cooperative Agreement No,
CR-810581   by  the  Ohio  State
University under  the sponsorship of the
U.S. Environmental Protection Agency.
   4.0 z.	
     0 ?            8              15
    Days After Inoculation with Antagonists

Figure 2.   Population development of Fia-
           vobactenum balustinum 299ft
           f—	) and T. hamstum 382
           ( 	  I in a container medium
           planted with cucumber and
           amended with compost  F.
           baiustinum and T. hamatum
           were inoculated individually in
           a container medium amended
           with (1) conducive compost from
           the high temperature center of
           the curing pile { 01; (2) svppres-
           stve edge compost { B I; of 13)
           were introduced as a combina-
           tion (reaiment in the conducive
           center compost medium ( © )
           Vertical bars indicate standard
           error.

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  H.AJ. Hoitink, and A.F Schmrtthenner are  with the  Ohio State University, Ohio
        Agricultural Research and Development Center, Woosier, OH 44891
  James A. Ryan is the EPA Project Officer (see below).
  The  complete report, entitled  "Effects  of Composted  Municipal Sludge  on
        Soiihorne  Plant Pathogens," (Order  No.  PB  88  195  714-AS; Cost
        $14.95, subject to cfhange) will be avtitabte only from:
           National Technical Information Service
           5?85 Port Hoya/ Road
           Spring field, VA22161
           Telephone: 703-4S7-4SSO
  The EPA Project Officer can be contacted at:
           Water Engineering Research Laboratory
           U.S. Environmental Protection Agency
           Cincinnati, OH 45268
United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH
Official
Penalty for Private Use $300

EPA/6QO/S2-88/024

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