Insecticidal Effects of the Insect Growth Regulators

Methoprene and Pyriproxyfen on the Cat Flea

(Siphonaptera: Pulicidae)

HITOSHI KAWADA AND MASACHIKA HIRANO

Agricultural Chemicals Research Laboratory, Sumitomo Chemical Company Limited, Takarazuka,
Hyogo 665, Japan

J. Med. Entomol. 33(5): 819-822 (1996)

ABSTRACT Residual effectiveness of the insect growth regulators pyriproxyfen and meth-
oprene against cat flea, Ctenocephalides felis (Bouch6), larvae were evaluated under simulated
household conditions. Pyriproxyfen provided control of larvae for >12 mo when applied at 1
mg and 0.2 mg/m2 and S3 mo at the rate of 0.04 mg/m2. Methoprene applied at 1 mg/m2
provided control for >12 mo and 6 mo at 0.2 mg/m2. No significant mortality was observed
when methoprene was applied at 0.04 mg/m2.

KEY WORDS cat flea, pyriproxyfen, methoprene, juvenile hormone mimic, insect growth
regulator

THE CAT FLEA, Ctenocephalides felis (Bouche"), is
a major pest of humans and companion animals
because they are more synanthropic and have a
wider host range than other flea species. The lar-
vae are found in carpets and other areas of the
house and high populations of larvae are also found
in home yards, which provide a constant source of
flea infestation (Palma and Meola 1990). The lar-
vae appear to be the best life stage to target for
control because they are often distributed in the
pets bedding area (Osbrink et al. 1986).
Insect growth regulators (IGRs), such as juvenile
hormone mimics. or chitin synthesis inhibitors,
have been reported to be effective control agents
for flea larvae (El-Gazzar et al. 1986, Marchiondo
et al. 1990, Palma and Meola 1990, Henderson and
Foil 1993, Palma et al. 1993, Hinkle et al. 1995).
Pyriproxyfen (Nylar or Sumilarv) acts as a juvenile
hormone mimic and interrupts nymphal-adult or
pupal-adult metamorphosis (Hatakoshi et al. 1987,
1988). Its potential as a biorational insecticide cov-
ers a wide range of insect species of public health
or household importance, such as synanthropic
flies (Kawada et al. 1987, Langley et al. 1988, Mil-
ler 1989, Bull and Meola 1993), mosquitoes (Ka-
wada et al. 1988, Mulla and Darwazeh 1988,
Schaefer et al. 1988, Suzuki et al. 1989, Kamimura
and Arakawa 1991, Okazawa et al. 1991), chiron-
omid midges (Ali et al. 1993, Trayler et al. 1994),
cockroaches (Kawada et al. 1989, Koehler and Pat-
terson 1991), termites (Su and Scheffrahn 1989),
and fleas (Palma and Meola 1990, Hinkle et al.
1995). Palma and Meola (1990) reported that Ny-
lar, an emulsifiable concentrate formulation of
pyriproxyfen, used at the rate of 32 mg/m2 pre-
vented development of >80% cat flea larvae for 3
wk in home yard conditions. Total-release aerosol
0022-2585/96/0819-0822$02.00/0 © 1996 Entomological Society of America
formulation of pyriproxyfen used at the rate of
>0.39 mg/m2 (0.01% [Al] wt:wt for 0.18-liter aero-
sol can) significantly reduced adult flea emergence
by >80% for a >7-mo duration (Hinkle et al.
1995). In similar experiments using total-release
aerosols, we reported that 0.91 mg/m2 of pyripro-
xyfen (0.0375% [Al] wt:wt for a 0.18-liter aerosol
can) prevented the emergence of the cat flea for
>7 mo in the laboratory, and that 2.4 mg/m2 of
pyriproxyfen controlled a cat flea population > 1
mo in private houses infested with the cat flea
(Senbo et al. 1993). The objective of this study was
to compare the residual effectiveness of spray for-
mulation of pyriproxyfen and methoprene under
simulated household conditions.
Materials and Methods
Test Compounds. A 5% emulsifiable concen-
trate formulation of pyriproxyfen (4-phenoxyphen-
yl (RS)-2-(2-pyridyloxy) propyl ether [Sumitomo,
Takarazuka, Hyogo, Japan]) and methoprene (iso-
propyl-(2E,4E)-ll-methoxy-3,7,ll-trimethyl-2,4-
dodecadienoate [Sandoz, Des Plaines, IL]) was
used to prepare the dilution of test sprays.
Test Insects. Second-instar cat flea larvae were
used as test insects. The fleas were collected in
Sakai, Osaka, Japan, in 1991, reared with cats as a
host for several generations at Osaka Seiyaku (Hi-
gashi Osaka, Japan) and introduced to our labora-
tory in 1993. The larvae were reared in a medium
that contained a mixture of 80 g sand, 15 g pow-
dered animal food (CE-2, Clea Japan, Tokyo), 3 g
dried yeast (Ebios, Tanabe Pharmaceutical, To-
kyo), and 2 g dried bovine blood (Wako, Tokyo).
The strain showed normal susceptibility to pyre-
820 JOURNAL OF MEDICAL ENTOMOLOGY Vol. 33, no. 5

Table 1. Residual effectiveness of ICRs on carpet under simulated household conditions

Dosage, % survival at month after treatment

mg/m2 0 1 2 3 4 5 6 9 12
Control 93.3a 90.0a 93.3a 76.7a 93.3a 76.7a 96.7a 85.0a 83.3a
Pyriproxyfen 1 0b 3.33b 0b 3.33bc 0b 0b 0b 3.33b 0b
0.2 0b 10.0b 0b 0c 0b 0b 3.33b 6.67b 0b
0.04 10.0c 6.67b 0b 30.0ab 10.0b 66.7a
Methoprene 1 10.0c 6.67b 0b 0c 6.67b 0b 10.0b 16.7b 6.67b
0.2 36.7c 16.7c 3.33b 6.67bc 13.3b 16.7b 23.3b 50.0a
0.04 53.3ac 50.0c 50.0c 53.3ab 73.3a — — — —

Numbers followed by the same letter are not significantly different (P = 0.01; Tukey HSD test).

throids and carbamates and slight tolerance to or- completed. Adult emergence was observed, and in-
ganophosphates (Kobayashi et al. 1994a). hibition of emergence was determined by compar-
Evaluation Methods. Polyester carpet (0.4 cm ison with untreated test samples. Bioassays were
deep) was cut into square pieces (15 by 15 cm) conducted every 4 wk for 12 mo.
and glued to plywood backing. Twelve carpet sam-
ples were prepared for each IGR concentration. A
5% emulsifiable concentrate formulation of the Results and Discussion
IGR was diluted with water to the appropriate con- The untreated control survival was 76.7-96.7%
centration and the carpet was treated at 50 ml/m2. during the 12-mo period. Larval survival in the
Each sample was vacuumed with 4 strokes by a
pyriproxyfen treatment was significantly lower than
vacuum cleaner (MC-104H, Matsushita Electric,
Tokyo) and then 1 g of dried animal food (CE-2, that in the control for >12 mo at concentrations
Clea Japan) was distributed uniformly onto the of 1 and 0.2 mg/m2, and for 5:3 mo at 0.04 mg/m2
surface of the carpet. The carpets were maintained (Table 1). The effective duration of the metho-
at 25°C and 60% RH. A series of each concentra- prene treatments was for >12 mo at 1 mg/m2 and
tion of the test samples was prepared each week. for 6 mo at 0.2 mg/m2. No significant difference in
Three pieces of round plugs (5.5 cm diameter) larval survival was obtained even immediately after
were cut from each carpet test sample and then treatment when fleas were exposed to the carpet
placed in an aluminum cup (5.5 cm diameter, 5.5 samples treated with methoprene at 0.04 mg/m2.
cm high). Ten 2nd-instar fleas were released onto The LD50 values of pyriproxyfen and methoprene
each plug and the cup was covered with a plastic during the 12-mo period are shown in Table 2. The
lid that contained 4 small ventilation holes. A piece LD50s were constantly <0.04 mg/m2 by 4 mo and
of adhesive paper (3 by 3 cm) was stapled on the 0.04-0.2 mg/m2 by 12 mo after exposure of the flea
inside wall of the aluminum cup to trap adults that larvae to pyriproxyfen. The LD 50 of methoprene
emerged. The cups were stored in a desiccator that tended to increase gradually monthly. Hinkle et al.
provided 80—90% RH until adult emergence was (1994) reported that the survival of cat flea larvae
exposed to commercial total-release aerosol for-
mulation of methoprene (mixture of 0.075% [S]-
Table 2 . Estimated LD50 of pyriproxyfen and meth- methoprene, 1.0% propoxur, 0.47% dichlorvos) be-
oprene residues aged on carpet came insignificant when compared with the control
by the 3rd mo and that 0.025% pyriproxyfen
Month LD 50 (mg/m2) at month after treatment (95% FL)
after snowed a significant reduction in the survival by
treat- Pyriproxyfen Methoprene the 7th mo. In their experiment the concentrations
ment of 0.075% [S]-methoprene and 0.025% pyriproxy-
0 <0.04 0.076
fen were calculated to be SS2.41 mg/m2. This
amount is equivalent to CS4.82 mg/m2 of metho-
1 <0.04 0.047 prene because [S]-isomer is thought to be twice as
2 <0.04 0.043
active as racemic methoprene (Chamberlain et al.
(-) (0.019-0.046) 1988) and 0.975 mg/m2, respectively. Our results
3 <0.04 0.062 indicate a significant difference in the effective
(-) (0.038-0.078) dosages for pyriproxyfen and methoprene between
4 <0.04 0.087 the treatment with total-release aerosol and that of
(-) (0.023-0.13)
5 0.04-0.2 0.04-0.2 emulsifiable concentrate. Actual amounts of IGR
in the carpet appear to be much higher using the
6 0.04-0.2 0.026 emulsifiable concentrate spray treatment than the
total-release aerosol treatment. Moreover the for-
9 0.04-0.2 0.28
(0.044-0.36)
mulation factors, such as inner pressure, liquid/gas
(-)
12 0.04-0.2 0.2-1.0 ratio, particle size, and discharge rate, may affect
the actual amount of IGR on the floor when the
September 1996 KAWADA AND HIRANO: EFFECTS OF
total release aerosol treatment is used. The relative
activity ratio of pyriproxyfen to methoprene was
approximately >5 times in the different treat-
ments. These results indicate the higher persis-
tence of pyriproxyfen on the carpet under house-
hold conditions than methoprene, although the
relative activity of pyriproxyfen had been reported
to be less than twice the activity of methoprene
(Kobayashi et al. 1994b). Hinkle et al. (1995) at-
tributed the lack of residual effectiveness in the
methoprene treatment to its high volatility, be-
cause the structure of methoprene is similar to hy-
droprene and is reported to translocate or move
from the point of application (Donahue and Young
1992). Their conclusions, however, are not consis-
tent with the vapor pressure of the 2 compounds
within the same range of 2.37 X 10~5 mmHg at
25°C for methoprene (Farm Chemical Handbook
1995) versus 2.8 X 10- 5 mmHg at 20°C for pyri-
proxyfen (Preiss and Untiedt 1994). Volatility,
therefore, is not a factor by which the difference
in residual activities of 2 chemicals can be ex-
plained. High stability of chemicals in the carpet
matrix or in the house dust is more likely to be the
reason for the long-term residual effectiveness in
the carpet treatments. Diflubenzuron (Uniroyal,
Amsterdam), a chitin synthesis inhibitor with a rel-
atively high stability like pyriproxyfen, was report-
ed to have a long residual effectiveness under sim-
ulated household conditions (Henderson and Foil
1993). The dosage of diflubenzuron, however, was
much higher (53.82 mg/m2) than the dose of pyr-
iproxyfen used in the current study. Such a high
dosage of diflubenzuron was required because of
its lower inhibitory activity than the other IGRs
such as methoprene (Moser et al. 1992).
Carpets seem to be one of the more difficult
substrates to be treated effectively with an insec-
ticide because of their increased surface area (Hin-
kle et al. 1995). Osbrink et al. (1986) reported the
lack of residual effectiveness of pressurized aero-
sols containing pyrethrins and tetramethrin against
cat flea larvae. They also found that the combina-
tion of IGRs with the chemicals noted above re-
inforced their residual effectiveness. The use of
IGRs with high stability and high inhibitory activ-
ity, such as pyriproxyfen, would minimize the total
amount of chemicals used in houses and reduce
the operational cost in the household flea control
program.
Acknowledgment
We thank S. Senbo (Sumika Life Tech Company Lim-
ited, Osaka, Japan) for rearing and providing the experi-
mental insects.
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Received for publication 13 December 1995; accepted
19 April 1996.