Journal of Invertebrate Pathology 77, 198 –205 (2001)

doi:10.1006/jipa.2001.5015, available online at http://www.idealibrary.com on

The Effects of an Entomopathogenic Fungus, Beauveria bassiana

(Balsamo) Vuillemin (Hyphomycetes), on Prostephanus truncatus (Horn)
(Col.: Bostrichidae), Sitophilus zeamais Motschulsky
(Col.: Curculionidae), and Grain Losses in Stored Maize
in the Benin Republic
W. G. Meikle,* ,1 A. J. Cherry,* N. Holst,† B. Hounna,* and R. H. Markham‡
*Plant Health Management Division, International Institute of Tropical Agriculture, 08 B.P. 0932 Tri-Postal, Cotonou, Benin;
†Department of Crop Protection, Danish Institute of Agricultural Sciences, Flakkebjerg, 4200 Slagelse, Denmark; and ‡Plant Health
Management Division, International Institute of Tropical Agriculture, P.M.B. 5320, Ibadan, Nigeria
E-mail: wmeikle@ars-ebcl.org

Received September 11, 2000; accepted March 6, 2001

stores in sub-Saharan Africa (Hodges et al., 1983;

A fungal entomopathogen, Beauveria bassiana, was
used to treat maize ears placed in traditional grain
stores against Prostephanus truncatus in a field experi-
ment conducted from September 1997 to March 1998 in
the Benin Republic, West Africa. Treatments included
oil-based spray with and without conidia, maize stored
with and without the husk, and stores with and without
artificial infestation. Additional treated ears kept in in-
sect-proof cages under field conditions were sampled
weekly and exposed to insects to estimate the virulence
and persistence of the pathogen during the storage sea-
son. P. truncatus densities were significantly lower in
treatments that included conidia, although densities
were high in all artificially infested treatments and
grain losses were severe. The effect of the pathogen was
modeled with an exponential decay function and incor-
porated in a published P. truncatus simulation model.
The effects of hypothetical pathogens with different vir-
ulence and persistence characteristics were evalu-
ated in terms of insect density and percentage grain
loss. © 2001 Academic Press
Key Words: Prostephanus truncatus; Sitophilus zea-
mais; Teretrius nigrescens; Beauveria bassiana; stored
products; biological control; microbial control; spore
persistence.
INTRODUCTION
Prostephanus truncatus (Horn) (Coleoptera: Bos-
trichidae) is an important pest of traditional maize
1
To whom correspondence should be addressed at USDA, Agricul-
tural Research Service, European Biological Control Laborarory,
Campus International de Baillarguet, CS 90013 Montferrier Surlez,
34988 Saint Gely du Fesc Cedex, France.
Dick, 1988; Markham et al., 1994). In these stores
maize is usually kept on the cob and in the husk
(Dick, 1988). Recent surveys conducted by the Inter-
national Institute of Tropical Agriculture (IITA) in
Benin have shown that a large proportion of farmers
treat their maize stores prophylactically with insec-
ticides (Meikle et al., 2000). One of the principal
difficulties with use of grain protectants in tradi-
tional stores is that they are intended for admixture
with shelled grain and therefore may be less effective
when applied as a surface treatment to whole ears
stored with the husk in the traditional way. Further-
more, pesticides tend to remain where they were
originally applied, and if this is on the external
leaves of the maize husk, insects that have already
established themselves on the grain within the husk
may not come into contact with pesticide. Ento-
mopathogenic fungi, on the other hand, have the
potential to persist in the environment through sec-
ondary cycling and may be redistributed within a
store by the movement of the insects themselves,
although the activity of the fungi in the storage
environment may be limited by factors such as low
humidity.
A successful pathogen would exhibit sufficiently high
virulence against P. truncatus, with little or no ten-
dency to attack the introduced natural enemy of P.
truncatus, Teretrius (formerly Teretriosoma) nigre-
scens (Lewis) (Coleoptera: Histeridae), to prevent a
severe outbreak. The pathogen should be persistent
enough in the stored product system to control poten-
tial outbreaks for at least several weeks into the stor-
age season. The pathogen should also have the capacity
for transmission from an initial inoculum to and be-

0022-2011/01 $35.00 198

Copyright © 2001 by Academic Press
All rights of reproduction in any form reserved.
 EFFECTS OF B. bassiana ON P. truncatus AND STORED MAIZE 199

TABLE 1
Summary of Treatments in the Field Experiment
Infested by P. Presence Maize per No.
No. Treatment truncatus of husk replicate (kg) Storage structure reps.

Experiment 1
1 Conidia ⫹ husk Yes Yes 50 Basket grain store 4
2 Conidia ⫺ husk Yes No 50 Basket grain store 4
3 Oil only Yes Yes 50 Basket grain store 4
4 No treatment Yes Yes 50 Basket grain store 4
5 No infestation (or treatment) No Yes 50 Basket grain store 4

Experiment 2
6 Conidia No (only at sampling) Yes 5 Insect-proof cage 4
7 No treatment No (only at sampling) Yes 5 Insect-proof cage 4

tween invading individuals and to subsequent gene-
rations following decay of the original inoculum.
Bourassa (1998) reported a strain of the entomopatho-
genic fungus Beauveria bassiana (Balsamo) Vuillemin
(Hyphomycetes) that met the criteria for virulence and
specificity.
The experiment reported in this article was con-
ducted at IITA’s Plant Health Management Division in
Calavi, Benin, West Africa as part of a wider project to
evaluate the potential of entomopathogens as alterna-
tives to chemical insecticides or as adjuncts to T. ni-
grescens for management of P. truncatus. B. bassiana
was evaluated as a control agent against P. truncatus
in a 7-month field experiment involving 20 grain
stores, with the fungi being applied as conidia sus-
pended in oil (Lomer et al., 1997) to artificially infested
ears.
In addition to the field evaluation of a biopesticide
for stored products, simulation models were used to
extrapolate our results to situations not encountered in
the field. Simulation models are practical tools often
helpful in developing integrated pest management
(IPM) strategies. Dobie (1974), Maier et al. (1996),
Arthur et al. (1998), and Meikle et al. (1999) used
models of Sitophilus zeamais (L.) (Coleoptera: Curcu-
lionidae) population dynamics to evaluate the effects of
different management strategies. Flinn and Hagstrum
(1990) simulated the effects of management practices
on Rhyzopertha dominica (F.) (Coleoptera: Bostrichi-
dae) populations. Thomas et al. (1996) examined the
effects of fungal pathogens on locusts and grasshoppers
in West Africa. Modeling in stored-product systems is
not limited to insect population dynamics. Holst et al.
(2000) linked P. truncatus density to grain weight loss,
and this relationship, together with a P. truncatus
simulation model developed by Meikle et al. (1998),
was used in the present study to provide an estimate of
the impact of the biopesticide on reducing grain weight
loss under a range of conditions.
The experiments were constructed and executed at
the IITA Benin campus from September 1996 to March
1997. Experiment 1, consisting of treatments 1 to 5,
was intended to compare the effects of treatment either
with conidia of B. bassiana formulated in an oil carrier,
with an oil carrier alone, or with no treatment on P.
truncatus and S. zeamais populations in grain stores
(Table 1). Experiment 2, consisting of treatments 6
(treated with conidia) and 7 (control), was intended to
measure the virulence and persistence of the original
fungal inoculum under field conditions.
METHODS AND MATERIALS
Maize. Maize ears of a local cultivar ‘Gbogbe’ were
harvested at the end of July, allowed to dry in the field
for several weeks to lower the grain moisture content
for storage, and subjected to fumigation (Phostoxin)
under large plastic sheets to eliminate any insect in-
festation from the field. A 3-week period was allowed
for pesticide residues to disperse. Ears were sampled
immediately prior to the experiment to check for insect
infestations and none were found.
Production, formulation, and application of conidia.
Bourassa (1998) had identified B. bassiana isolate
IMI330194 as one of a series of isolates pathogenic to P.
truncatus. Isolate IMI330194 was originally collected
from Hypothenemus hampei Ferr. (Coleoptera: Scolyti-
dae) in Kenya and was selected for this study because
of its superior production characteristics. Conidia were
produced by use of a standard two-stage system (Jen-
kins et al., 1998) with rice as the solid substrate. Ex-
tracted and dried conidia were sieved with a 106-␮m
sieve to remove large hyphal fragments. Dried conidia
powder contained 4.59 ⫻ 10 10 conidia/g and had a ger-
mination rate above 98%. Conidia were applied to ears
in treatments 1, 2, and 6 at a dose rate of 2 ⫻ 10 10
conidia/kg of ears in an oil suspension containing 100 g
conidia/liter of a 70% kerosene:30% peanut oil blend
200 MEIKLE ET AL.
(Lomer et al., 1997) via a Micron “Ulva” hand-held
rotary sprayer (Micron Sprayer Ltd., Bromyard, UK).
Conidia were formulated on the day of application.
In preparation for treatment, ears were individually
laid side by side in 3-m rows in a terrace protected from
wind. Treatment 3, containing blank oil plus Lumogen
UV tracer (Hays Chemicals, London, UK), was applied
first. Half of the blank oil was applied to the exposed
side of the ears. All ears were then turned over and the
remainder of the blank oil applied. After application,
10 ears, randomly selected, were examined under ul-
traviolet light to assess the extent of spray coverage.
Treatments 1, 2, and 6 were similarly applied, but
without the addition of Lumogen. Spore germination
immediately after applications was evaluated by appli-
cation of formulation diluted in peanut oil onto Sab-
ouraud dextrose agar in petri dishes and incubation for
24 h at 25°C. Maize was placed in grain stores on the
same day that treatments were applied.
Maize storage and sampling: Experiment 1. In Ex-
periment 1, maize ears were stored in basket-type
grain stores (50 kg ears per store). One store repre-
sented one replicate and each treatment was replicated
four times. Stores were constructed at sites throughout
the IITA campus, with treatments randomly assigned
to stores. Care was taken to minimize treatment inter-
actions by placing the stores as distant from each other
as possible (minimum 50 m) and at least 5 m from any
road. P. truncatus adults were introduced to treat-
ments 1, 2, 3, and 4 at the rate of 100 insects (2/kg) per
store immediately after the stocking of each store. The
introduction of such a large number of insects was
intended to ensure that sufficient insects would remain
after the first two sampling occasions. The sex ratio of
the 1600 insects collected from standard laboratory
cultures for placement in stores was assumed to be 1:1
(Scholz, 1997; Shires, 1979), and age distribution was
assumed to be random.
The sample unit for Experiment 1 was the maize ear.
Sampling began 2 weeks after the start of the experi-
ment and continued thereafter at 1-month intervals for
a total of seven sampling occasions. For each grain
store sample, 10 ears were randomly selected from the
surface of the store and to a depth of ca. 20 cm, taking
care to select ears that were not touching each other
and keeping each ear separate for subsequent evalua-
tions. Selected ears were dehusked when necessary,
shelled, and sieved. The live and dead P. truncatus, T.
nigrescens, and S. zeamais were counted and percent-
age weight loss was measured with the count and
weigh technique (Boxall, 1986). Grain moisture con-
tent was measured as the change in weight of 3 g of
kernels after 24 h at 70°C and was replicated three
times per store sample. The weights of the individual
ears were used to convert mean insect densities to per
kg of dry, undamaged grain (Holst et al., 2000) to
remove the confounding effects of grain weight loss
during the course of the season. Dead insects of all
species were washed in distilled water and incubated
in petri dishes on moist filter paper for at least 10 days
to determine rates of sporulation, which were taken as
an indication of fungal infection. Densities of sporulat-
ing and nonsporulating P. truncatus and S. zeamais
cadavers were calculated as above.
Treatments 1, 2, 3, and 4 were compared with re-
spect to log 10(x ⫹ 1)-transformed total numbers of live,
dead, and sporulating P. truncatus per ear with an
ANOVA repeated-measures model (SAS, 1997), with
the sample replicate being the store. Treatments 1 and
2 (those treatments with conidia) were compared to
treatments 3 and 4 (those treatments without) by
planned contrasts. Treatments 1, 2, 3, 4, and 5 were
also compared with respect to log 10(x ⫹ 1)-transformed
total numbers of live and dead S. zeamais by the same
ANOVA model.
Maize storage and sampling: Experiment 2. In
treatments 6 and 7, treated but uninfested maize was
held in wire-mesh-covered cages, kept in grain stores
similar to those in Experiment 1, so that inoculum was
subjected to ambient field conditions while excluding
insects. One store was constructed per treatment, and
each store held four cages of 5 kg maize each. Sampling
began immediately after treatment application and
continued weekly for 25 weeks. For each sample, 1 ear
per cage was removed and placed in a separate wire
mesh cage. Fifty adult P. truncatus were placed on
each ear and the cages returned to the field store. After
2 weeks, ears were destructively sampled, grain mois-
ture content was measured as above (three samples per
treatment), and the number of live and dead insects
were counted and treated as described above for Ex-
periment 1. The mortality of P. truncatus individuals
and the density of sporulating individuals used in
treatments 6 and 7 were calculated as a proportion of
the 50 insects used in each replicate.
Modeling. Population simulation models of P. trun-
catus (Meikle et al., 1998) and S. zeamais (Meikle et al.,
1999) were used to evaluate the observed and potential
roles of the entomopathogens in this system. The mod-
els were driven by daily minimum and maximum tem-
perature, obtained from the weather station on the
IITA campus, and daily grain moisture content, which
was calculated by use of linear interpolation between
sampling occasions. The P. truncatus model was pa-
rameterized at the rate of 2 insects/kg at the time of
stocking (100 insects/50-kg store) with a background
immigration rate of 0.01 insects/kg/day (P. truncatus
were common at the experimental site). The actual
immigration rate was unknown but this rate has pro-
vided a good fit for simulating the observed population
dynamics of pests in other stores at this site. The
 EFFECTS OF B. bassiana ON P. truncatus AND STORED MAIZE
proportion of sporulating P. truncatus adults measured
in each sample of treatment 6 was considered equiva-
lent to the proportion of insects dying due to fungal
infection and the data were fitted to an exponential
decay function
R t ⫽ R 0 exp 冉⫺t 䡠 ln共0.5兲
h
冊 ,
where R t is the proportion of the adult population
killed on day t, R 0 is the proportion killed on the day
that the mortality factor first takes effect (the initial
kill rate), and h is the half-life of the mortality factor.
This function was applied only to adult insects in the
P. truncatus model.
The P. truncatus model (without the pathogen) also
calculated the number of adults/kg dying of old age
each day. The cumulative total of dead adults/kg was
used as an estimate of the maximum density of cadav-
ers, and, by multiplication of the daily cumulative total
of dead adults by a coefficient, the simulated total was
fit to cadaver densities observed in treatment 4. The
coefficient represented the proportion of cadavers of
one day surviving to the next day. With the decay
function described above, the half-life of a cadaver was
calculated.
To parameterize the immigration rate of the S. zea-
mais model, the change in S. zeamais density, aver-
aged over all treatments, between day 1 and the first
sampling occasion on day 13, was divided by 12 for a
daily immigration rate. The developmental rate of S.
zeamais under these conditions precludes the possibil-
ity of egg-to-adult development in fewer than 15 days
(Meikle et al., 1999), so all insects observed on day 13
were known to be immigrants.
FIG. 1. Average densities of live P. truncatus/kg for treatments
1–5 of a field experiment conducted from September 1997 to April
1998 at IITA, Calavi, Benin.
201
FIG. 2. Average densities of dead P. truncatus/kg for treatments
1–5 of a field experiment conducted from September 1997 to April
1998 at IITA, Calavi, Benin.
RESULTS AND DISCUSSION
Mean P. truncatus density (per kilogram), and mean
densities of sporulating and nonsporulating dead in-
sects per ear are shown for Experiment 1 (Figs. 1–3).
Average P. truncatus densities in the treatments with
fungal conidia were significantly lower than those
without and were significant with respect to linear,
quadratic, and cubic trends (Table 2). Nevertheless,
densities of P. truncatus in all artificially infested
treatments were very high within 4 months of the start
of the experiment, and stores in treatments 1
(conidia ⫹ husk), 3 (oil only), and 4 (no treatment)
achieved densities in excess of 3500 insects/kg (natural
infestations in excess of 2000 insects/kg have been
observed for P. truncatus [see Meikle et al., 1998]). The
densities observed here were far beyond those accept-
FIG. 3. Average densities of P. truncatus cadavers/kg which sub-
sequently sporulated under laboratory conditions for treatments 1–5
of a field experiment conducted from September 1997 to April 1998 at
IITA, Calavi, Benin.
202 MEIKLE ET AL.

TABLE 2
F Ratios of Repeated-Measures ANOVA of Treatments 1, 2, 3, and 4 with Respect to log 10(x ⫹ 1)-Transformed
Total P. truncatus Numbers per 10-Ear Sample
No. P. truncatus per 10 ears

Factor N.d.f. D.d.f. Alive Dead Spor.

Treatment 3 12 11.17* 8.87* 1.60

T1 ⫹ T2 vs T3 ⫹ T4 1 12 24.32* 11.85* 3.50
Date 6 72 228.40* 89.05* 3.13*
Treatment ⫻ Date 18 72 7.02* 3.98* 1.24
Linear 1 72 1204.90* 476.59* 6.37*
Quadratic 1 72 54.75* 9.22* 0.87
Cubic 1 72 228.82* 9.27* 0.01

Note. “Spor.” indicates the number of cadavers that subsequently sporulated under laboratory conditions; N.d.f. and D.d.f. indicate
numerator and denominator degrees of freedom, respectively.
* Significant at P ⬍ 0.05.

able to farmers, and indeed no differences of practical In Experiment 2, insect mortality in treatment 6
importance were observed between the treated and the (conidia) was, on average, about 36% higher than that
untreated grain stores. Average P. truncatus densities in treatment 7 (control) for the first 49 days (Fig. 4).
in treatment 2 (conidia ⫺ husk) reached over 900 in- This period corresponds to the peak activity period of
sects/kg by the fifth sampling occasion and then de- the fungus in treatment 6, as measured by sporulation
clined to a level much lower than those in treatments 1, and, since cadavers were not surface sterilized, it
3, or 4. The number of dead insects recovered in treat- should be considered the maximum estimate. As mor-
ment 2 also declined, suggesting that factors other tality in treatment 6 declined, presumably due to de-
than adult mortality were responsible. Treatment 2 creasing quantities of viable spore residues, so did mor-
was the only treatment in which the kernels them- tality in treatment 7. No cadavers from treatment 7
selves were treated with the conidia. Population dy- sporulated, other than a small number infected by the
namics observed in treatment 3 were similar to those saprophytic fungi Aspergillus spp. and Fusarium spp.
in treatments 1 and 2 which included conidia. P. trun- Furthermore, mortality for both treatments 6 and 7
catus densities in treatment 5 (no infestation or treat- increased again, after the spore viability could be as-
ment) never became severe. No cadavers were found sumed to be negligible. This suggests that other factors
externally sporulating under field conditions, and played an important role in insect mortality. Grain
sporulation data pertains to cadavers which sporulated moisture content decreased throughout the season,
under humid conditions in the laboratory. The sporu- and the two lowest grain moisture contents recorded,
lation in treatments 3 and 4 may indicate the back- 10.5 and 12.5%, were both associated with peaks in P.
ground infection rate (in high-density situations) or
pathogens which had spread with the hosts to the other
stores. Because cadavers were washed with distilled
water only, all sporulation data should be considered
maximum estimates.
Densities of S. zeamais, which was the only other
primary pest found in the stores, were on average
lower than those for P. truncatus (the maximum mean
densities did not exceed 1000 insects/kg for any treat-
ment). These densities are similar to those that have
been observed previously in West Africa (Meikle et al.,
1999). The most sporulating S. zeamais cadavers, 10.6/
kg, were observed in treatment 2 (10 individuals were
found in a single ear). Although T. nigrescens was not
found in any store until the third sampling occasion
and did not exceed 2/kg until the sixth sampling occa-
sion (152 days after stocking), their densities exceeded
30/kg in all artificially infested treatments by the end FIG. 4. Average proportion of dead and sporulating P. truncatus
cadavers, from a group of 50 insects per sample, in treatments 6 and
of the experiment. Few dead T. nigrescens were found, 7 of a field experiment conducted from September 1997 to April 1998
and none sporulated. The effects of T. nigrescens on at IITA, Calavi, Benin. All sporulation was observed under labora-
P. truncatus density were not considered further. tory conditions.
 EFFECTS OF B. bassiana ON P. truncatus AND STORED MAIZE
FIG. 5. Exponential decay curve fit through raw data on propor-
tion of sporulating cadavers in treatment 6 of a field experiment
conducted from September 1997 to April 1998 at IITA, Calavi, Benin,
as shown in Fig. 4.
truncatus mortality in both treatments. However, Bell
and Watters (1982) showed that the minimal grain
moisture content necessary for P. truncatus larval sur-
vival is about 9% and probably lower for adults.
The exponential decay function fit the sporulation
data from treatment 6 adequately (adjusted r 2 ⫽ 0.73;
R 0 ⫽ 0.62 with SE ⫽ 0.034) and showed a half-life of
about 16.6 days (SE ⫽ 1.600) (Fig. 5). Model outputs
with and without the mortality factor differed, partic-
ularly during the first three sampling occasions, but
these differences would have been difficult to detect at
such low densities without larger samples (Fig. 6).
Overall, the model without the mortality factor simu-
lated densities observed in treatments 1, 3, and 4 well,
and the model with the mortality factor underesti-
mated observed densities in those treatments for most
of the season. Both models overestimated densities in
treatment 2 at the end of the season. Although the
immigration rate, 0.01 insects/kg/day, was considered
low, omitting the immigration rate considerably re-
duced the fit of the model to the field data.
The mortality factor was incorporated in the model
as a daily “kill rate.” In treatment 6 we assumed that
transmission from the substrate to the insect was
100% (insects were placed directly on the treated ears)
and that the kill rate was determined largely by viru-
lence and persistence. The transmission rate is of par-
ticular interest here because the ability of the patho-
gen to cause disease outbreaks over a long period of
time is postulated to be a major advantage of a patho-
gen over a pesticide. In this study cadavers were ob-
served to sporulate, a necessary step for secondary
cycling, only after being placed under conditions of
high humidity. It is possible that internal sporulation
may have occurred, in which case spores would have
been released only when the cadavers disintegrated.
203
The release rate of viable spores, which would be ex-
pected to affect the transmission rate, is a function of
the disintegration rate of the cadavers and the persis-
tence of the spores within the cadavers. The daily dis-
integration rate of cadavers per day as a proportion of
available cadavers was estimated to be 0.63, and indi-
vidual cadavers were estimated to have a half-life of
about 0.70 days (with an assumed adult life expectancy
of 60 days [Shires, 1979]). These estimates apply to
cadavers of uninfected insects; the disintegration rate
of infected cadavers may be different. The disintegra-
tion rate was assumed to be constant during the course
of the season (although it may change as densities of
secondary pests and saprophytic organisms change)
and dead and dying insects were assumed to remain in
the ear and not to leave or fall out. Little is known
about the persistence of fungus within insect cadavers
in field situations. Internal sporulation is likely to oc-
cur when external conditions are unfavorable, and
there is evidence to suggest that, in the case of control
of locusts and grasshoppers in the Sahel by Metarhi-
zium anisopliae var. acridum, it may contribute to the
long-term persistence of fungal inoculum in otherwise
harsh environments (Thomas et al., 1996).
The significantly lower density of P. truncatus in
stores treated with conidia compared to untreated
stores and the sporulation of cadavers collected from
grain stores well after the period of peak fungal activ-
ity give rise to the possibility that transmission leading
to secondary cycling may have occurred. Of the 119
cadavers recovered, 92% were found on sampling occa-
sions four, five, and seven. The large number of sporu-
lating cadavers collected on sampling occasion seven,
was largely due to the 35 cadavers found on a single
ear in treatment 1. If the ear with 35 cadavers is
excluded, 72% of the cadavers were recovered in sam-
FIG. 6. Simulated P. truncatus densities, with and without the
mortality factor shown in Fig. 5, compared to average densities of
live P. truncatus observed in a field experiment conducted from
September 1997 to April 1998 at IITA, Calavi, Benin.
204 MEIKLE ET AL.
pling occasions four and five, and most cadavers in all
treatments were recovered on sampling occasions four
and five. Although no more than 45 sporulating
cadavers recovered from any artificially infested store,
and therefore the possibility that all cadavers belonged
to introduced insects infected by the original fungal
inoculum cannot be excluded, the estimated cadaver
half-life and the low probability of finding so many
such cadavers across all treatments within such a
short period of time suggest that some transmission
occurred.
Another explanation for the significantly lower P. trun-
catus density in stores treated with conidia is the impact
of the fungus on larval P. truncatus. This isolate of B.
bassiana caused very high and rapid mortality among P.
truncatus larvae in laboratory tests (Bourassa, 1998).
Dead larvae would have been difficult to identify and
were not counted. A period of high larval mortality would
be one way to simulate the population dynamics observed
in treatment 2 and would also account for the lack of dead
adult insects as the adult density decreased in that treat-
ment. The incorporation of a larval-stage transmission
factor into the simulation model was not seen as practical
in this study, since it would require several assumptions
and would be impossible to validate without additional
field data. However, with a better understanding of the
relationship of insect density and weather to factors such
as sporulation rates, spore production per cadaver, and
insect movement within the store, incorporating trans-
mission into a model would present little difficulty.
Model results with and without the mortality factor
calculated from the observed parameters were com-
pared to results with versions of the mortality factor
calculated from modified parameters. The modifica-
tions included (1) increased initial kill rate, R 0 , from
FIG. 7. Simulated P. truncatus densities per kilogram, with (sol-
id line) and without (dotted line) the mortality factor and with
modified mortality factors: a greater R 0 of 0.8 (short-dashed line), a
longer half-life h of 30 days (dashed and dotted line), and both a
greater R 0 and a longer h (long-dashed line).
FIG. 8. Simulated densities compared to observed average den-
sities of live S. zeamais in a field experiment conducted from Sep-
tember 1997 to April 1998 at IITA, Calavi, Benin.
0.62 to 0.8 of the population, (2) increased half-life, h,
from 16.6 days to 30 days, and (3) increased R 0 and h
(Fig. 7). The modifications taken singly did not have
much effect on the simulated P. truncatus population
dynamics, although together they showed a pro-
nounced effect.
Since P. truncatus and S. zeamais were by far the most
common insect pests in the grain stores, grain weight
losses were estimated with simulation models of each
species (Holst et al., 2000). The immigration rate for the
S. zeamais simulation model was estimated at 0.8 in-
sects/kg/day. Simulated S. zeamais densities overesti-
mated observed densities for the last four sampling occa-
sions, and average S. zeamais declined among treat-
ments 1 through 5 in the last two sampling occasions
(Fig. 8). The reasons for this decline are unknown; com-
petition with P. truncatus is unlikely to be involved since
the decline was also observed in treatment 5 with a
comparatively low P. truncatus density.
Grain weight losses exceeded 55% in treatments 1, 2,
3, and 4 by the end of the experiment, far in excess of
what is acceptable to farmers. Grain from stores with
losses above 15% is difficult to sell, and the mean losses
among treatments 2, 3, and 4 exceeded that level
within 4 months. Simulated grain weight loss, by use of
models without pathogens, compared well with ob-
served values, although losses for the last sampling
occasion were slightly underestimated (Fig. 9). Simu-
lated losses with a P. truncatus model with the patho-
gen effect underestimated losses much more. By inclu-
sion of the mortality factor with increased virulence
and persistence in the insect model, expected losses in
this high-density situation would not have exceeded
15% until the 6th month.
Further development of an effective stored-product bio-
pesticide will involve identification of an acceptable ap-
plication technique and carrier (kerosene, used here as
 EFFECTS OF B. bassiana ON P. truncatus AND STORED MAIZE
FIG. 9. Simulated percentage weight losses, calculated from sim-
ulated P. truncatus (with and without a mortality factor) and S.
zeamais densities shown in Figs. 6 and 8, compared to average
weight losses observed in a field experiment conducted from Septem-
ber 1997 to April 1998 at IITA, Calavi, Benin.
part of the carrier formulation, should never be used to
treat food) and selection of more effective pathogen iso-
lates. A better understanding of sporulation behavior and
the mechanism of horizontal and vertical transmission in
grain stores will help us to understand how isolates with
particular characteristics may perform as biopesticides.
Simulation modeling might, in due course, be useful for
selection of the most promising isolates for further field
testing, after preliminary bioassays have been carried
out, by showing how a “profile” of characteristics might
combine in the action of a biopesticide. Simulation mod-
eling, as it was conducted here, was useful in organizing
information and clarifying assumptions about pathogen
activity, and it may have a role in clarifying the role of
biopesticides within a comprehensive IPM strategy for
stored product pests in Africa.
ACKNOWLEDGMENTS
We thank S. Korie, J. Langewald, C. Nansen, P. Neuenschwander,
F. Schulthess, and two anonymous reviewers for helpful comments.
The research was conducted at the Plant Health Management Divi-
sion of the International Institute of Tropical Agriculture and sup-
ported by the Danish International Development Agency (DANIDA)
and the German Agency for Technical Cooperation (GTZ).
REFERENCES
Arthur, F. H., Throne, J. E., Maier, D. E., and Montross, M. D. 1998.
Feasibility of aeration for management of maize weevil popula-
tions in corn stored in the southern United States: Model simula-
tions based on recorded weather data. Am. Entomol. 44, 118 –123.
Bell, R. J., and Watters, F. L. 1982. Environmental factors influenc-
ing the development and rate of increase of Prostephanus trunca-
tus (Horn) (Coleoptera: Bostrichidae) on stored maize. J. Stored
Products Res. 18, 131–142.
Bourassa, C. 1998. “Utilisation de Champignons Entomopathogènes
Contre le Grand Capucin du Maı̈s, Prostephanus truncatus (Horn)
205
(Col.: Bostrichidae), en Afrique de l’Ouest.” MSc thesis, University
of Quebec at Montreal.
Boxall, R. A. 1986. “A Critical Review of the Methodology for Assess-
ing Farm-Level Grain Losses after Harvest.” Report of the Tropical
Development and Research Institute, G191.
Dick, K. 1988. “A Review of Insect Infestation of Maize in Farm Storage
in Africa with Special Reference to the Ecology and Control of Pro-
stephanus truncatus.” Overseas Development Natural Resources In-
stitute Bulletin 18, NRI, Chatham Maritime, Kent, UK.
Dobie, P. 1974. The laboratory assessment of the inherent suscepti-
bility of maize varieties to postharvest infestation by Sitophilus
zeamais Motsch. J. Stored Products Res. 10, 183–197.
Flinn, P. W., and Hagstrum, D. W. 1990. Simulations comparing the
effectiveness of various stored-grain management practices used
to control Rhyzopertha dominica (Coleoptera: Bostrichidae). Envi-
ron. Entomol. 19, 725–729.
Hodges, R., Dunstan, W. R., Magazini, I., and Golob, P. 1983. An
outbreak of Prostephanus truncatus (Horn) (Coleoptera: Bostrichi-
dae) in East Africa. Protect. Ecol. 5, 183–194.
Holst, N., Meikle, W. G., and Markham, R. H. 2000. Grain injury
models for Prostephanus truncatus (Coleoptera: Bostrichidae) and
Sitophilus zeamais (Coleoptera: Curculionidae) in rural maize
stores in West Africa. J. Econ. Entomol. 93, 1338 –1346.
Jenkins, N. E., Heviefo, G., Langewald, J., Cherry, A. J., and Lomer,
C. J. 1998. Development of mass production technology for aerial
conidia for use as mycopesticides. Biocontrol News Inform. 19,
21N–31N.
Lomer, C. J., Prior, C., and Kooyman, C. 1997. Development of
Metarhizium spp. for the control of grasshoppers and locusts.
Mem. Entomol. Soc. Can. 171, 265–286.
Maier, D. E., Adams, W. H., Throne, J. E., and Mason, L. J. 1996.
Temperature management of the maize weevil, Sitophilus zeamais
Motsch. (Coleoptera: Curculionidae) in three locations in the
United States. J. Stored Products Res. 32, 255–273.
Markham, R. H., Bosque-Pérez, N. A., Borgemeister, C., and Meikle,
W. G. 1994. Developing pest management strategies for Sitophilus
zeamais and Prostephanus truncatus in the tropics. FAO Plant
Protect. Bull. 42, 97–116.
Meikle, W. G., Holst, N., and Markham, R. H. 1999. A simulation model
of Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae) in
rural grain stores in Benin. Environ. Entomol. 28, 836 – 844.
Meikle, W. G., Holst, N., Degbey, P., and Oussou, R. 2000. An
evaluation of sequential sampling plans for the larger grain borer
(Coleoptera: Bostrichidae) and the maize weevil (Coleoptera: Cur-
culionidae) and of visual grain assessment in West Africa. J. Econ.
Entomol. 93, 1822–1831.
Meikle, W. G., Holst, N., Scholz, D., and Markham, R. H. 1998.
Simulation model of Prostephanus truncatus (Coleoptera: Bos-
trichidae) in rural maize stores in the Republic of Benin. Environ.
Entomol. 27, 59 – 69.
SAS Institute Inc. 1997. SAS/STAT Software: Changes and En-
hancements, through Release 6.12, SAS Institute Inc., Cary, NC.
Scholz, D. 1997. “Dispersal and Host-Finding Behaviour of Pro-
stephanus truncatus (Horn) (Col.: Bostrichidae).” Ph.D. thesis,
University of Hanover, Germany.
Shires, S. W. 1979. Influence of temperature and humidity on sur-
vival, development period and adult sex ratio in Prostephanus
truncatus (Horn) (Coleoptera, Bostrichidae). J. Stored Products
Res. 15, 5–10.
Thomas, M. B., Langewald, J., and Wood, S. N. 1996. Evaluating the
effects of a biopesticide on populations of the variegated grasshop-
per, Zonocerus variegatus. J. Appl. Ecol. 33, 1509 –1516.