VETERINARY ENTOMOLOGY
Evaluation of Three Formulations of Beauveria bassiana for Control
of Lesser Mealworm and Hide Beetle in Georgia Poultry Houses
CHRISTOPHER J. GEDEN1
AND
DONALD C. STEINKRAUS2
USDAÐARS, Center for Medical, Agricultural and Veterinary Entomology, P.O. Box 14565, Gainesville, FL 32604
KEY WORDS Alphitobius diaperinus, Dermestes maculatus, Beauveria bassiana, poultry
THE LESSER MEALWORM, Alphitobius diaperinus (Panzer), and hide beetle, Dermestes maculatus DeGeer,
are among the most important pests of the poultry
industry throughout the world (Safrit and Axtell 1984,
Turner 1986, Geden et al. 1999). A. diaperinus is a
reservoir of numerous avian and human pathogens,
and their larvae are highly destructive pests in building
insulation and other soft construction materials
(McAllister et al. 1994, 1996; Vaughan et al. 1984;
Despins et al. 1994; LeTorcÕh and Letenneur 1983).
Damage to building materials occurs when larvae
reach critical population levels in the litter or manure,
resulting in dispersal of mature larvae seeking pupation sites (Ichinose et al. 1980, Geden and Axtell 1987).
Hide beetles are also commonly found in poultry
houses, where they feed on cracked chicken eggs,
dead birds and rodents, and manure itself (Cloud and
Collison 1986, Stafford et al. 1988). Hide beetle larvae
damage buildings in a manner similar to that of A.
diaperinus and, in addition, also tunnel into structural
timbers to the point of compromising the soundness of
building supports. Annual economic losses to these
insects, known collectively as litter beetles, have been
E-mail: cgeden@gainesville.usda.uß.edu.
Department of Entomology, University of Arkansas, Fayetteville,
AR 72701.
1
2
estimated at $16 million and $10 million in Virginia and
Georgia, respectively (Turner 1986, Riley et al. 1997).
Because insecticidal control of litter beetles is generally unsatisfactory and short-lived, there is a need
for alternative management approaches. Mechanical
barriers can be used to prevent beetles from emigrating from the manure to reach susceptible building
components (Geden and Carlson 2001), but this approach does not limit populations living in the manure.
The beetles have several natural enemies, including
mites and protozoans (Husband and Baker 1992,
Steinkraus and Cross 1993, Steinkraus et al. 1992,
Apuya et al. 1994, Bala et al. 1990). Lesser mealworms
are also susceptible to steinernematid and heterorhabditid nematodes, which can provide short-term control in the Þeld (Geden et al. 1985, 1987, Geden and
Axtell 1988).
In previous studies we found that larvae, pupae, and
adults of A. diaperinus were susceptible to the entomopathogenic fungus B. bassiana, and that there were
substantial differences in virulence among fungal
strains (Steinkraus et al. 1991, Geden et al. 1998).
Although direct treatment of infested poultry litter
and manure is not the most effective delivery method,
this is a simple application method that is likely to be
used by commercial producers. The objective of the
current study was to determine the effectiveness of
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J. Econ. Entomol. 96(5): 1602Ð1607 (2003)
ABSTRACT Initial screening of 12 Beauveria bassiana (Balsamo) Vuillemin isolates against larvae
of the lesser mealworm (Alphitobius diaperinus [Panzer]) resulted in the selection of two isolates,
GHA and 707, for further testing under Þeld conditions. Three formulations of each strain were
prepared: an EC, a ground corn granular formulation, and waste product of fungal propagation
containing spent media, mycelia, and unharvested conidia (“residue” formulation). Two Þeld trials
were conducted in commercial caged-layer houses in Georgia with 5Ð 6 mo of manure accumulation
and established populations of A. diaperinus and hide beetles (Dermestes maculatus DeGeer). In the
Þrst trial Þeld, B. bassiana was applied a single time to the manure surface at either 109 (EC and granular
cornmeal bait formulations) or 108 (residue formulation) fungal spores per square meter. In the second
trial, two successive weekly treatments were applied, using a total of 6X the rate of application used
in the Þrst trial, SigniÞcant treatment effects were short-lived and only detected 2 wk after treatment
in both trials. The granular formulations of both strains and the residue formulation of the GHA strain
provided the greatest degree of suppression (60 Ð90%) of beetle larvae. A laboratory bioassay conÞrmed that the granular bait was the most effective formulation. More frequent applications made
earlier in the manure accumulation cycle may be necessary to achieve satisfactory control of these
beetles.
October 2003
GEDEN AND STEINKRAUS: EVALUATION OF THREE FORMULATIONS OF B. bassiana
manure treatments with different strains and formulations of B. bassiana for control of established litter
beetle populations in commercial poultry houses.
Materials and Methods
the houses were delineated by placing ßags along the
sections of manure ⬇2-m wide by 5 m long. Manure
had accumulated for ⬇6 mo at the beginning of the
study. Two tube traps (Safrit and Axtell 1984) were
placed in each plot and adult and larval beetles of A.
diaperinus and D. maculatus were collected from the
traps and counted for three weeks before assigning
treatment groups to the plots. The two plots with the
lowest larval A. diaperinus counts were dropped, leaving 28 plots to allocate among the six fungal treatments
and controls (four plots/treatment). Plots were allocated by Þrst ranking them by counts of A. diaperinus
larvae and grouping them into four sets of seven plots
(i.e., plots no. 1Ð7 in abundance of beetle larvae, 8 Ð14,
etc.), in descending order of larval abundance. Treatments were assigned randomly within each group of
seven plots until all 28 plots had been assigned. By
doing this, plots were assigned to treatments so that
there were no signiÞcant pretreatment differences in
A. diaperinus larval densities among the treatment
groups.
B. bassiana was applied to the manure plots on 31
August, 2000, at 109 spores per square meter for the EC
and granular formulations. The residue formulation
was applied at a lower dose (108 spores per square
meter) because of the lower spore concentration inherent in this product. The EC was applied by diluting
50 ml of concentrated spore suspensions in 1000 ml of
water for each plot and spraying the manure with a
calibrated CO2 sprayer. The two dry formulations
were applied by hand with conventional restaurant
grated cheese shakers. Beetle populations were monitored weekly with tube traps as before for 3 wk after
treatment.
A second trial was conducted in a different poultry
house at the Hoboken farm in SeptemberÐNovember,
2000. Manure in the house had accumulated for ⬇5 mo
at the start of testing. Methods were the same as
described above, except that two applications of B.
bassiana were made, 1 wk apart, at three times the
spore dose used in the Þrst trial. Also, pretreatment
counts were made for 4 wk before allocation to treatment groups. Treatments were applied on 26 October
and 2 November. Beetle populations were monitored
weekly for 3 wk after the second treatment.
Data were analyzed by one-way analysis of variance
(ANOVA) separately for each week after performing
log (x ⫹ 1) transformations of trap counts, and treatment means were separated using the REGWQ algorithm of the GLM Procedure of the Statistical Analysis
System (SAS Institute 1998).
Laboratory Bioassays. Laboratory assays were conducted in Arkansas using the same rates of application
used in the Þrst Þeld trial (six fungal treatments plus
controls). In these tests, groups of 100 Þeld-collected
larvae were Þrst placed in screen-topped 3-liter plastic
tubs containing 800 cm3 of poultry litter from a broiler
house. Litter was frozen before testing to kill any
insects present at the time of collection. Treatments
were applied to the litter surface 1-h after addition of
the larvae and the tubs held at 25⬚C for three weeks.
Chicken feed was added to the tubs as needed and
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Strain Selection Bioassay. Twelve isolates of B. bassiana maintained by Mycotech Co. (Butte, MT) were
screened for virulence against larvae of A. diaperinus
in the laboratory: 706, 707, 708, 709, 711,712, GHA,
1201a, 1201b, 1202a, 1202b, 1202c. Spores were harvested from agar plates of live culture material, then
spore counts per mg of collected spores were determined by hemacytometer counts. Spore counts
ranged from 1.2 ⫻ 107 (isolate 1202c) to 1.2 ⫻ 108
(isolate 706) spores/mg.
Spore viability was determined for each isolate by
diluting 1 mg of spores in 1 ml of distilled water
containing 0.01% Tween 80, making a 1:10 dilution,
then plating 0.1 ml on a petri dish containing SDA agar
(two dishes/isolate). After 24 h, four coverslips were
placed on each plate over a drop of lactophenol-acid
fuschin stain, and the number of germinated and ungerminated spores in one Þeld of view under each
coverslip was determined at 400⫻. Spore viability
ranged from 62.0 to 99.1%.
Spores were weighed and mixed with corn starch to
prepare two concentrations of viable spores for each
isolate; 1.5 ⫻ 107 and 1.5 ⫻ 109 spores/g of starch.
Forced-contact bioassays were then conducted separately in our laboratories in Arkansas and Florida.
Spore mixtures were tested against mature A. diaperinus larvae by exposing groups of 10 larvae to 0.1-g
aliquots of the test mixtures per group. After a 30-s
period of contact, the larvae were removed from the
inocula with soft forceps and transferred to clean petri
dishes lined with Þlter paper premoistened with distilled water. Cornstarch with no spores was used for
controls. Larvae were monitored for mortality on days
3, 5, and seven after exposure. The bioassays were
replicated either four (Arkansas) or Þve times (Florida), using beetle larvae collected from poultry houses
in Arkansas and Georgia, respectively. Data from the
two laboratories were pooled and two isolates were
selected for production, formulation, and Þeld evaluation.
Field Trials in Georgia. Three formulations of the
two selected isolates (GHA and 707) were prepared
by Mycotech for Þeld-testing. The Þrst was an EC
formulation containing 2 ⫻ 1011 spores/ml. The second was a coarsely ground granular corn bait consisting of 8.45 ⫻ 109 spores/g. The third formulation,
“residue,” was a waste product of fungal propagation
comprised of spores and mycelia remaining in spent
fungal growth medium after automated spore harvesting at MycotechÕs production facility. This product,
which was processed to the consistency of coarse
cornmeal, contained 6.5 ⫻ 108 spores/g.
Field tests were conducted at a commercial poultry
farm near Hoboken, GA, consisting of four layer
houses containing ⬇120,000 birds each. In the Þrst
trial, 30 ⬇10-square-meter plots of manure in one of
1603
1604
JOURNAL OF ECONOMIC ENTOMOLOGY
Table 1. Relative virulence of 12 strains of B. bassiana for
field-collected mature lesser A. diaperinus larvae after a brief
forced-contact exposure to spores mixed with corn starch
Mean (SE) percent mortality of A. diaperinus after
exposure to B. bassiana
Strain
Day 7 after exposure
to dose
1.5 ⫻ 106
1.5 ⫻ 108
1.5 ⫻ 106
1.5 ⫻ 108
10.0 (10.0)
14.5 (2.5)
8.5 (5.5)
5.5 (2.5)
6.5 (3.5)
49.0 (49.0)
60.0 (40.0)
3.5 (0.5)
6.0 (6.0)
5.0 (5.0)
4.0 (4.0)
13.5 (9.5)
5.0 (5.0)
33.5 (10.5)
60.5 (12.5)
36.5 (1.5)
41.0 (3.0)
16.0 (6.0)
100.0 (0.0)
100.0 (0.0)
95.0 (5.0)
76.5 (0.5)
10.0 (10.0)
9.0 (5.0)
23.5 (13.5)
5.0 (5.0)
67.0 (33.0)
28.5 (18.5)
54.0 (16.0)
58.5 (24.5)
84.0 (16.0)
96.5 (3.5)
100.0 (0.0)
38.5 (18.5)
60.5 (26.5)
20.5 (13.5)
61.5 (31.5)
63.0 (37.0)
10.0 (10.0)
95.0 (5.0)
77.0 (23.0)
83.0 (17.0)
86.0 (14.0)
97.0 (3.0)
100.0 (0.0)
100.0 (0.0)
99.0 (1.0)
98.5 (1.5)
60.5 (2.5)
80.0 (20.0)
87.0 (13.0)
10.0 (10.0)
Doses are spores per 0.1 g of corn starch used in the assays.
N ⫽ 4 (Arkansas) and 5 (Florida) replications, 10 larvae per strain
and dose.
Results are means of tests replicated in Florida using Georgia beetle
populations and in Arkansas using local beetle populations.
moisture levels maintained by periodically measuring
moisture loss from the tubs and adding water to compensate. Larvae were removed by hand from onethird of the tubs at 1, 2, and 3 wk after treatment and
numbers of live and dead larvae counted (N ⫽ 4
tubs/treatment/time interval for a total of 84 tubs).
Data were analyzed by ANOVA and treatment means
separated weekly as before.
Results and Discussion
All of the isolates tested were virulent for A. diaperinus larvae in forced-contact bioassays (Table 1).
Table 2.
The low concentration of 1.5 ⫻ 107 spores/g of corn
starch was selected because this was the approximate
LC50 observed using this bioassay method in a previous B. bassiana evaluation against this pest (Geden et
al. 1998). The 707 isolate was the most virulent in these
tests (100% mortality at the low dose on day 7 and
100% mortality at the high dose on day 5) and was,
therefore, selected for the Þeld tests. The GHA strain
ranked near the middle of the isolates tested in virulence for beetle larvae; however, it was included in the
Þeld trials because this isolate possesses desirable
characteristics with respect to production yield, harvesting efÞciency, and storage stability. Moreover, a
product containing this isolate (Mycotrol) is already
registered for use against whiteßies, aphids, thrips,
mealybugs, leafhoppers, weevils, and leaf-feeding insects in a variety of agricultural systems.
In the Þrst Þeld trial, the only signiÞcant treatment
effects on A. diaperinus larvae were observed at 2 wk
after treatment (Table 2), when the granular formulations of both isolates and the residue formulation of
GHA resulted in ⬎50% reduction in the number of
larvae collected in tube traps (means of 15.5Ð18.3
larvae/trap) compared with the controls (40.3 larvae/
trap) (F ⫽ 3.32; df ⫽ 6,21; P ⫽ 0.019). This suppression
was short-lived, however, and no signiÞcant treatment
effects were observed on week 3. Variation in adult A.
diaperinus counts was high and prevented detection of
signiÞcant treatment effects at P ⫽ 0.05 (Table 2).
However, as was observed with the larvae of this
species, there were numerically fewer adult beetles
collected from plots treated with the two granular
formulations and the residue formulation of GHA
(means, 28.0 Ð 43.3 adults/trap) compared with the
control plots (129.8 adults/trap).
Hide beetle populations, although relatively small,
were affected by the treatments in much the same
way as A. diaperinus (Table 3). Treatment effects on
Mean (SE) A. diaperinus per tube trap before and after manure treatment with B. bassiana in trial no. 1
Weeks
posttreatment
B. bassiana strain 707 formulation
EC
Granular
⫺3
⫺2
⫺1
Pre-trt mean
0
1
2
3
5.5 (2.9)a
4.0 (1.1)a
30.3 (10.1)a
13.2a
12.5 (4.3)a
26.8 (12.9)a
24.5 (5.4)ab
31.0 (6.0)a
3.0 (1.1)a
9.0 (5.0)a
35.5 (19.5)a
15.8a
27.0 (8.8)a
31.3 (12.6)a
15.5 (5.5)b
51.3 (24.8)a
⫺3
⫺2
⫺1
Pre-trt mean
0
1
2
3
36.0 (8.6)a
49.3 (33.4)a
150.5 (66.5)a
78.6a
144.8 (73.7)a
104.5 (60.3)a
86.0 (31.5)a
150.3 (92.6)a
37.3 (16.0)a
38.8 (14.2)a
105.3 (28.9)a
60.5a
93.0 (58.1)a
58.3 (24.9)a
43.3 (9.4)a
73.0 (28.5)a
Residue
B. bassiana strain GHA formulation
EC
Control
Granular
Residue
A. diaperinus larvae
5.5 (1.0)a
5.8 (2.8)a
17.0 (7.0)a
6.0 (2.5)a
16.8 (5.3)a
25.5 (7.0)a
13.1a
12.4a
55.3 (28.1)a
49.3 (18.3)a
18.8 (3.8)a
62.0 (14.6)a
33.5 (10.6)ab
59.3 (8.0)a
59.5 (39.7)a
29.5 (6.1)a
7.0 (2.3)a
10.3 (4.9)a
22.0 (11.6)a
13.1a
7.3 (2.2)a
8.0 (2.9)a
18.0 (5.1)b
27.0 (6.7)a
7.0 (3.2)a
18.5 (5.6)a
13.3 (4.4)a
12.9a
41.0 (8.8)a
69.5 (27.3)a
18.3 (4.3)b
38.3 (15.7)a
2.8 (2.1)a
2.3 (0.5)a
32.5 (11.4)a
12.5a
38.3 (7.2)a
26.0 (6.0)a
40.3 (15.6)a
29.5 (8.9)a
A. diaperinus adults
42.3 (8.5)a
25.8 (7.3)a
78.5 (20.3)a
35.8 (13.2)a
85.3 (22.3)a
91.8 (39.3)a
68.7a
51.1a
92.3 (31.5)a
113.3 (40.5)a
50.8 (7.0)a
68.5 (16.9)a
89.8 (49.2)a
83.8 (24.3)a
234.0 (183.4)a
66.5 (22.9)a
64.0 (30.8)a
32.0 (11.8)a
72.5 (24.5)a
56.2a
32.3 (18.0)a
20.3 (9.7)a
28.0 (4.6)a
42.3 (2.3)a
33.0 (8.1)a
84.8 (48.1)a
90.0 (40.8)a
69.2a
70.8 (21.9)a
80.5 (17.5)a
37.0 (8.9)a
117.0 (71.3)a
37.5 (11.7)a
31.5 (12.8)a
131.8 (19.8)a
66.9a
33.5 (4.5)a
31.0 (12.2)a
129.8 (75.8)a
73.3 (22.1)a
Treatments applied on 31 Aug. (week 0) to 10 m2 manure plots at 109 (EC and granular) or 108 (residue) B. bassiana spores per m2. N ⫽
4 plots/treatment, two tube traps/plot. Means within rows followed by the same letter are not signiÞcantly different at P ⫽ 0.05 (REGWQ).
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1201a
1201b
1202a
1202b
1202c
706
707
708
709
711
712
GHA
control
Day 5 after exposure
to dose
Vol. 96, no. 5
October 2003
Table 3.
GEDEN AND STEINKRAUS: EVALUATION OF THREE FORMULATIONS OF B. bassiana
1605
Mean (SE) D. maculatus per tube trap before and after manure treatment with B. bassiana in trial no. 1
EC
Granular
⫺3
⫺2
⫺1
Pre-trt mean
0
1
2
3
11.5 (3.7)a
4.5 (2.6)a
24.8 (10.3)a
13.6a
18.3 (7.4)a
8.5 (2.5)a
15.3 (3.7)a
10.8 (6.3)a
12.0 (4.6)a
8.0 (3.1)a
33.0 (13.0)a
17.7a
6.3 (1.6)a
7.5 (2.9)a
2.0 (0.0)bc
1.0 (0.7)a
D. maculatus larvae
13.8 (4.2)a
24.5 (10.2)a
8.0 (1.3)a
15.8 (6.6)a
20.0 (5.8)a
40.8 (13.2)a
13.9a
27.0a
10.5 (3.4)a
23.0 (6.8)a
9.8 (6.3)a
22.0 (7.2)a
5.5 (2.3)ab
10.8 (5.7)ab
2.3 (0.9)a
7.5 (3.1)a
⫺3
⫺2
⫺1
Pre-trt mean
0
1
2
3
0.5 (0.5)b
1.0 (0.7)a
5.5 (1.8)a
2.3a
6.5 (3.0)a
4.3 (1.4)a
4.3 (1.3)a
5.5 (2.9)a
1.5 (0.6)ab
1.8 (1.2)
12.0 (9.4)a
5.1a
1.8 (0.9)a
2.5 (1.2)a
1.5 (1.0)ab
0.0 (0.0)b
D. maculatus adults
2.8 (0.6)ab
8.8 (3.7)ab
3.3 (0.9)a
7.3 (3.7)a
7.0 (6.0)a
18.8 (13.5)a
4.4a
11.6a
1.5 (0.6)a
8.0 (5.2)a
3.5 (1.4)a
8.0 (4.6)a
0.5 (0.3)ab
2.3 (0.5)ab
0.0 (0.0)b
2.8 (0.9)ab
B. bassiana strain BB-707 formulation
Residue
B. bassiana strain GHA formulation
EC
Granular
Residue
11.0 (4.0)a
3.8 (2.8)a
13.8 (5.4)a
9.5a
8.3 (5.0)a
5.8 (2.3)a
1.3 (0.9)c
3.5 (1.3)a
10.5 (2.2)a
4.8 (1.3)a
12.5 (3.1)a
9.3a
5.3 (2.9)a
5.5 (0.9)a
1.3 (0.3)c
5.3 (3.0)a
3.3 (2.3)ab
1.0 (0.6)a
2.8 (1.5)a
2.4a
1.3 (1.3)a
1.3 (0.8)a
0.0 (0.0)b
0.0 (0.0)b
2.3 (1.0)ab
2.0 (0.7)a
5.5 (2.2)a
3.3a
2.3 (0.9)a
3.3 (1.7)a
0.0 (0.0)b
2.8 (2.1)b
Control
10.5 (5.5)a
8.8 (3.7)a
34.8 (17.0)a
18.0a
19.5 (16.8)a
12.8 (11.4)a
11.0 (6.4)a
7.0 (3.4)a
13.5 (5.7)a
4.3 (2.0)a
3.8 (2.2)a
7.2a
4.5 (3.6)a
5.3 (4.6)a
1.0 (1.0)ab
3.0 (1.2)ab
Treatments applied on 31 Aug. (week 0) to 10 m2 manure plots at 109 (EC and granular) or 108 (residue) B. bassiana spores per m2. N ⫽
4 plots/treatment, two tube traps/plot. Means within rows followed by the same letter are not signiÞcantly different at P ⫽ 0.05 (REGWQ).
larvae were only signiÞcant on week 2 after treatment,
with the granular formulation (both isolates) and
GHA residue plots having 12Ð18% as many larvae as
the control plots (F ⫽ 5.15, df ⫽ 6.21; P ⫽ 0.002).
Treatment effects on hide beetle adults were erratic, presumably because of the small numbers collected.
In the second Þeld trial, populations of A. diaperinus
were higher overall than in the Þrst trial (Table 4). The
only signiÞcant treatment effects on larvae were obTable 4.
Mean (SE) A. diaperinus per tube trap before and after manure treatment with B. bassiana in trial no. 2
Weeks posttreatment
(date)
EC
Granular
⫺4
⫺3
⫺2
⫺1
Pre-trt. mean
0
0
1
2
3
8.8 (1.4)a
15.0 (3.3)a
19.0 (8.2)a
46.5 (18.2)a
22.3a
36.3 (7.0)a
16.8 (4.8)ab
74.5 (9.3)abc
33.3 (4.8)a
23.0 (14.2)a
10.3 (3.2)a
5.0 (1.4)a
7.0 (2.9)a
49.8 (7.2)a
18.0a
62.3 (16.0)a
35.0 (28.0)ab
57.3 (10.0)bc
40.3 (12.3)a
40.5 (16.7)a
23.8 (6.9)a
37.0 (6.6)a
12.5 (3.0)a
61.8 (26.7)a
33.8a
86.5 (23.6)a
110.3 (33.0)a
229.8 (112.7)a
178.5 (75.7)a
96.5 (46.6)a
⫺4
⫺3
⫺2
⫺1
Pre-trt. mean
0
0
1
2
3
served on week 1 after the second treatment (2 wk
after the Þrst fungal application). As in the Þrst trial,
the two granular formulations and the GHA residue
plots had signiÞcantly lower beetle populations
(mean, 41Ð57.3 larvae/trap) compared with the controls (144.0 larvae/trap) (F ⫽ 6,36, df ⫽ 6,21; P ⫽
0.001), but the suppression was short-lived. No significant treatment effects were observed among A. diaperinus adults (Table 4) or hide beetle adults or larvae
in the second trial (Table 5).
B. bassiana strain BB-707 formulation
31.3 (5.6)a
51.8 (24.5)a
21.8 (6.2)a
14.5 (5.0)a
29.9a
62.8 (8.4)a
82.5 (7.3)a
187.5 (53.8)a
174.8 (48.1)a
160.8 (77.2)a
Residue
B. bassiana strain GHA formulation
EC
Control
Granular
Residue
A. diaperinus larvae
15.5 (3.2)a
10.5 (4.1)a
10.5 (1.0)a
4.5 (2.1)a
15.5 (6.3)a
3.5 (0.6)a
33.5 (3.7)a
42.0 (9.1)a
18.8a
15.1a
24.5 (4.4)a
51.8 (21.8)a
7.0 (1.4)ab
15.3 (5.7)ab
86.5 (12.8)abc
116.0 (30.8)ab
22.5 (6.6)a
44.8 (13.3)a
13.3 (5.5)a
25.3 (13.3)a
16.5 (9.0)a
23.8 (12.8)a
6.0 (3.7)a
41.8 (7.9)a
22.0a
66.0 (19.4)a
2.8 (1.3)b
41.0 (4.7)c
37.8 (13.0)a
40.0 (16.6)a
7.8 (1.5)a
5.8 (4.1)a
3.8 (1.9)a
60.0 (12.9)a
19.3a
20.5 (6.5)a
11.3 (0.6)ab
47.8 (8.0)c
15.8 (3.8)a
6.8 (2.6)a
2.8 (0.8)a
11.0 (4.0)a
10.8 (4.2)a
42.0 (10.2)a
16.6a
35.0 (5.7)a
38.3 (18.5)a
144.0 (13.4)a
82.5 (39.3)a
68.8 (30.0)a
A. diaperinus adults
34.3 (8.6)a
38.3 (11.5)a
93.5 (51.1)a
36.5 (8.1)a
64.0 (40.9)a
21.5 (1.9)a
46.3 (21.4)a
66.8 (40.0)a
59.5a
40.8a
70.8 (44.8)a
142.0 (107.9)a
110.8 (19.0)a
189.5 (119.7)a
191.3 (29.0)a
202.8 (48.8)a
125.0 (38.4)a
97.8 (16.5)a
36.0 (6.0)a
54.5 (41.9)a
31.5 (7.0)a
45.8 (16.9)a
22.3 (7.7)a
69.5 (33.6)a
42.3a
58.0 (17.2)a
168.0 (90.7)a
148.3 (30.5)a
252.0 (56.3)a
148.0 (139.4)a
32.0 (10.4)a
23.8 (7.4)a
15.8 (4.8)a
60.8 (33.6)a
33.1a
92.0 (25.2)a
192.3 (18.5)a
225.8 (44.7)a
113.3 (58.2)a
57.0 (29.0)a
32.3 (6.0)a
74.8 (26.1)a
34.5 (15.3)a
57.8 (39.0)a
49.9a
143.3 (44.1)a
147.8 (42.0)a
343.8 (89.4)a
237.5 (46.4)a
172.3 (80.4)a
Treatments applied on two consecutive weeks (26 Oct. and 2 Nov.) to 10 m2 manure plots at 3 ⫻ 109 (EC and granular) or 3 ⫻ 108 (residue)
B. bassiana spores/m2. N ⫽ 4 plots/treatment, 2 tube traps/plot. Means within rows followed by the same letter are not signiÞcantly different
at P ⫽ 0.05 (REGWQ).
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Weeks posttreatment
(date)
1606
JOURNAL OF ECONOMIC ENTOMOLOGY
Table 5.
Vol. 96, no. 5
Mean (SE) D. maculatus per tube trap before and after manure treatment with B. bassiana in trial no. 2
B. bassiana strain BB-707 formulation
EC
Granular
⫺4
⫺3
⫺2
⫺1
Pre-trt. mean
0
0
1
2
3
1.3 (1.3)
0.3 (0.3)
1.5 (0.9)
0.3 (0.3)
0.9
3.5 (1.0)
5.3 (3.1)
7.0 (4.0)
7.3 (2.0)
4.0 (2.1)
0.8 (0.5)
0.3 (0.3)
1.5 (0.6)
2.3 (1.1)
1.2
2.5 (0.3)
9.3 (1.8)
7.5 (3.2)
4.8 (3.4)
6.0 (5.7)
⫺4
⫺3
⫺2
⫺1
Pre-trt. mean
0
0
1
2
3
0.0 (0.0)
0.0 (0.0)
0.0 (0.0)
0.3 (0.3)
0.1
1.0 (1.0)
2.8 (2.4)
3.8 (2.1)
3.3 (2.0)
0.5 (0.5)
0.0 (0.0)
0.3 (0.3)
0.3 (0.3)
1.3 (0.6)
0.5
1.5 (1.2)
6.5 (2.9)
8.5 (6.0)
5.0 (3.0)
1.5 (1.5)
Residue
B. bassiana strain GHA formulation
EC
Control
Granular
Residue
D. maculatus larvae
2.3 (1.7)
0.8 (0.5)
1.0 (0.6)
0.5 (0.5)
2.5 (1.0)
1.0 (0.6)
1.8 (0.9)
1.0 (0.6)
1.9
0.8
7.8 (3.7)
4.3 (1.7)
7.8 (3.3)
5.8 (3.2)
8.3 (3.1)
5.3 (2.1)
10.0 (4.1)
4.5 (0.9)
2.3 (0.5)
5.3 (1.8)
6.3 (5.9)
4.0 (2.7)
8.8 (6.8)
2.8 (1.5)
5.5
3.8 (2.8)
4.0 (2.7)
6.8 (2.2)
12.0 (4.3)
1.8 (0.9)
2.5 (1.0)
0.5 (0.5)
3.5 (2.2)
0.8 (0.3)
1.8
4.0 (0.4)
10.3 (2.6)
6.5 (1.3)
7.0 (2.1)
3.3 (1.4)
0.0 (0.0)
0.3 (0.3)
3.3 (1.6)
1.0 (0.4)
1.2
7.8 (5.5)
6.8 (2.3)
8.5 (2.2)
8.8 (4.4)
4.0 (0.9)
D. maculatus adults
1.3 (0.6)
0.0 (0.0)
0.3 (0.3)
0.3 (0.3)
1.8 (1.4)
0.5 (0.3)
0.0 (0.0)
0.0 (0.0)
0.9
0.2
2.8 (1.8)
2.8 (2.1)
3.3 (2.0)
6.0 (3.5)
5.5 (3.0)
4.3 (2.5)
3.5 (1.7)
7.0 (1.7)
0.0 (0.0)
0.8 (0.5)
3.8 (3.8)
0.0 (0.0)
4.0 (3.7)
0.3 (0.3)
2.0
1.5 (0.6)
4.5 (3.1)
7.5 (5.2)
3.3 (1.1)
0.5 (0.5)
0.0 (0.0)
0.0 (0.0)
0.5 (0.5)
0.5 (0.3)
0.3
4.5 (2.2)
12.0 (8.1)
4.0 (2.3)
2.5 (2.2)
1.5 (1.2)
0.0 (0.0)
0.0 (0.0)
0.5 (0.5)
0.3 (0.3)
0.2
9.0 (8.3)
1.8 (1.1)
7.0 (2.9)
22.0 (19.0)
1.8 (0.9)
Treatments applied on two consecutive weeks (26 Oct. and 2 Nov.) to 10 m2 manure plots at 3 ⫻ 109 (EC and granular) or 3 ⫻ 108 (residue)
B. bassiana spores/m2. N ⫽ 4 plots/treatment, 2 tube traps/plot). There were no signiÞcant treatment effects on any week at P ⫽ 0.05
(REGWQ).
These Þeld trials represented rather strenuous test
conditions for the fungus. The manure pack was already large (5Ð 6 mo accumulation) and robust populations of A. diaperinus were already present at the
beginning of the trials. Given these circumstances and
the rapid burial of the applied spores under a constantly falling carpet of fresh manure deposits, our
tests may be seen as representing a worst-case scenario for B. bassiana poultry house applications. The
effects of manure conditions such as pH, moisture, and
feed additives on fungal efÞcacy are unknown. Nonetheless, the results indicate that reductions in larval
densities of both A. diaperinus and hide beetles can be
achieved with fungal treatments and that granular and
residue formulations are more effective in poultry
manure than an EC formulation. Further support for
Table 6. Mortality of A. diaperinus larvae in laboratory arenas
treated with B. bassiana using 109 (EC and granular) or 108
(residue) B. bassiana spores/m2
Strain
Control
707
707
707
GHA
GHA
GHA
Formulation
EC
Granular
Residue
EC
Granular
Residue
Mean (SE) percent mortality of A.
diaperinus larvae after exposure to
B. bassiana treatment
Week 1
Week 2
Week 3
5.1 (1.8)a
3.8 (0.5)a
5.2 (1.6)a
4.6 (2.7)a
7.8 (2.9)a
6.8 (3.1)a
8.1 (1.8)a
15.8 (3.0)c
28.1 (4.0)bc
57.2 (3.2)a
33.8 (4.2)b
26.8 (4.3)bc
28.5 (8.5)bc
19.3 (5.0)c
34.3 (5.0)c
45.2 (7.1)c
81.5 (6.6)a
48.7 (4.0)bc
41.5 (4.4)c
65.9 (9.8)ab
48.1 (5.7)bc
N ⫽ 4 arenas containing 800 cm3 of used broiler litter and an initial
population of 100 A. diapreinus larvae. Means within columns followed by the same letter are not signiÞcantly at P ⫽ 0.05 (REGWQ).
this is provided by results of the laboratory experiment, where the granular formulations provided
greater control than the EC (Table 6).
In summary, the results of the current study indicate
that single or two weekly treatments of B. bassiana
were not effective in providing long-term control of
established resident litter beetle populations in a large
manure mass. However, the results are sufÞciently
encouraging to warrant further evaluations under test
conditions more favorable to the fungus. For example,
weekly treatments during the Þrst few weeks after
manure clean out would increase the probability of
the fungus impacting the target population in time to
prevent the beetles from reaching outbreak levels.
Incorporation of food attractants or semiochemicals
into granular baits or waste residue products could
improve the efÞcacy of these materials. Carefully
timed applications of infective baits could also be
integrated with other bio- and behavior-based management approaches such as mechanical barriers,
mites, protozoans, and fungal treatments directed at
migrating larvae (Bala et al. 1990, Steinkraus et al.
1992, Steinkraus and Cross 1993, Apuya et al. 1994,
Geden et al. 1998, Geden and Carlson 2001).
Acknowledgments
The authors thank Richard DeMaio and Stefan Jaronski for
their support of this work and for providing material for
testing. Thanks also to Henry McKeithen and Haze Brown for
assisting with Þeld work. The research was supported in part
by a grant awarded to Mycotech, Co. from the USDA SBIR
program.
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Weeks posttreatment
October 2003
GEDEN AND STEINKRAUS: EVALUATION OF THREE FORMULATIONS OF B. bassiana
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1607
Christopher Geden