J. Agric. Food Chem.

1995, 43, 507-512 507

Neem Oil and Neem Oil Components Affect the Efficacy of

Commercial Neem Insecticides
John D. Stark*,+and James F. Walter'
Washington State University, Puyallup Research and Extension Center, 7612 Pioneer Way East,
Puyallup, Washington 98371, and W.R. Grace and Company, Washington Reseach Center,
7379 Route 32, Columbia, Maryland 21044

A series of toxicity studies with pea aphid, Acyrthosiphon pisum (Harris) and the neem insecticides
Margosan-0 (MO), MO devoid of neem oil, Azatin, RH-9999, Azatin with 5% neem oil, RH-9999
with 5%neem oil, and neem oil (5%)were conducted. We found that addition of neem oil increased
the efflcacy of neem insecticides that did not contain the oil, while removal of neem oil from MO
reduced its efficacy 62%. Neem oil was also extracted with methanol. When methanol-extracted
neem oil was added to MO devoid of neem oil, its efficacy was still 30% lower than MO. Addition
of canola oil gave a similar response. Six limonoids, nimbandiol, deacetylnimbin, 6-acetylnimbandiol,
deacetylsalannin, nimbin, and salannin, and two unidentified chemicals, believed t o be limonoids,
were identified in neem oil. Our data indicate that neem oil and other oils increase the efficacy of
neem insecticides, but a polar component(s) of neem oil also contributes to increased biological activity
of neem insecticides.

Keywords: Neem; oil; efficacy; insect control

INTRODUCTION interfered with molting and reduced longevity and

fecundity in a dose-dependent manner. The study
Environmentally benign pesticides, which we define presented here was originally conducted to compare the
as pesticides that are selectively toxic, do not bioaccu- toxicity of several neem insecticides to the pea aphid.
mulate, and exhibit relatively short persistence in the We expected to find little or no difference in toxicity
environment, are needed for modern integrated pest between three commercially developed neem insecti-
management programs. Neem insecticides appear to fit cides that contained azadirachtin as the active ingredi-
this definition because they have been shown to be ent. What we found was quite different and led us to
selective (Saxena et al., 1984; Mansour et al., 1987; Kale examine the relationship between neem oil and the
et al., 1986; Rossner and Zebitz, 1987; Stark et al., effectiveness of neem insecticides.
1992), have short persistence (Stark, unpublished re-
sults), and pose less negative impacts to ecosystems
than conventional insecticides (Stark, 1992). MATERIALS AND METHODS
Interest in neem insecticides has grown over the past Insects. Pea aphids were obtained from cultures main-
10 years as more pesticides are lost due to environmen- tained at Washington State University, Puyallup Research and
tal and food safety problems (Koul et al., 1990; Schmut- Extension Center, Puyallup, WA.
terer, 1990; Ascher, 1993). Two neem insecticides are Chemicals. The following neem insecticides were evalu-
now commercially available in the United States (Mar- ated: Margosan-0 (MO), an EC formulation which contains
gosan-0, W.R. Grace and Co., Columbia, MD, and 0.25%azadirachtin and 3-5% neem oil, formulated neem oil
Azatin, Agndyne Technologies Inc., Salt Lake City, UT) (90%AI EC), MO without neem oil which was specially made
and have recently received an exemption from residue for this study, and clarified neem oil, which is purified neem
oil that is unformulated (all from W.R. Grace and Co.,
tolerance on food crops by the U S . EPA. Several other Columbia, MD), Azatin, an EC formulation with 3% aza-
neem products may soon be available. There are dirachtin and no neem oil (Agridyne Technologies Inc., Salt
differences in formulation between the neem insecticides Lake City, UT), and RH-9999, a chemically modified (hydrox-
available in the United States. Margosan-0 (MO) has ylated) azadirachtin (20%AI wp) which also contains no neem
a 0.25%azadirachtin content and 3-5% neem oil while oil (Rohm and Haas Co., Philadelphia, PA). The azadirachtin
Azatin has 3% azadirachtin, but no neem oil. Both MO content of chemical standards was quantitated throughout the
and Azatin are emulsifiable concentrate formulations. study with the methods developed by Hull et al. (1993).
RH-9999, another neem insecticide in the experimental Experiment 1. Comparative Effects on Population
stage, produced by Rohm and Haas Co. (Philadelphia, Growth. In a previous study we determined that population
PA), is a wettable powder that contains chemically growth ofA. pisum reared on broad bean, Vicia faba L., which
modified (hydroxylated) azadirachtin (20% AI) and no had been treated with 100 mg of azadirachtifi was only 28%
of controls (Stark and Rangus, 1994). Following the same
neem oil. Neemguard (W.R. Grace and Co., Columbia, procedures presented in Stark and Rangus, we tested MO,
MD) is a formulated neem oil product from neem seed Azatin, and RH-9999 at the 100 mg of azadirachtifi level.
kernels (90%AI) which has insecticidal activity for some We used caged potted broad bean, V . faba L. var. Banner, and
species. aphids. Broad bean plants were planted seven per pot (10 cm
Stark and Rangus (1994) found that MO was toxic t o diameter) in potting medium. Plants and aphids were kept
the pea aphid, Acyrthosiphon pisum (Harris). MO *
in an environmental chamber at 25 =t0.5 "C and 78 5% RH
and a 16:8 light-dark regimen. When the plants were ca. 25
cm high, they were thinned to five per pot and sprayed t o
Washington State University. runoff with the equivalent of 100 mg of azadirachtifi of water
* W.R. Grace and Company. MO, RH-9999, or Azatin with a Thomas atomizer powered by
0021-8561/95/1443-0507$09.00/0 0 1995 American Chemical Society
508 J. Agric. Food Chem., Vol. 43, No. 2, 1995 Stark and Walter

Table 1. Final Population Density of A. pisum Exposed Table 3. Toxicity of Neem Insecticides to A. pisum
to Broad Bean Treated with Several Neem Insecticides Exposed as First Instars to Neem Insecticides at 100 mg
at the Equivalent of 100 mg of Azadirachtid of AzadirachtinlL or 5%Neem Oil (Experiment 3)"
(Experiment 1)"
% mortality f SD
no. of aphids i SD
Margosan-0 devoid
control Azatin MO RH-9999 control Margosan-0 of neem oil neem oil
1392.25 i 58.66a 654.75 i 41.3313 232.00 i 3 9 . 7 0 ~1378.50i 56.51a oc 92.5 f 5.00a 35.0 k 5.7713 oc
O1 ANOVA followed by lsd test. Means followed by the same a ANOVA followed by lsd test. Means followed by the same
letter are not significantly different. Based on four replicates. letter are not significantly different. Based on four replicates.
Table 2. Toxicity of Neem Insecticides to Immature A. with MO presented by Stark and Rangus (1994). The methods
pisum Exposed as First Instars to Neem Insecticides at used for topical toxicity tests are described in Stark and
100 mg of AzadirachtidL (Experiment 2Ia Rangus (1994).
L7c mortality i SD Experiment 6. Methanol Extraction of Neem Oil and
Addition of Canola Oil. To determine whether a component
control Margosan-0 Azatin RH-9999 of neem oil was responsible for the effects observed in the
2.0 & 4 . 4 7 ~ 90.0 f 7.07a 68.0 & 10.95b 8.0 & 1 3 . 0 4 ~ previous experiments or whether the oil itself imparted the
increase in toxicity observed, we compared MO, MO devoid of
a ANOVA followed by lsd test. Means followed by the same
neem oil, MO with methanol-extracted neem oil (5%),MO with
letter are not significantly different. Based on five replicates. canola oil (5%),and canola oil (5%). Canola oil was chosen
because, like neem oil, it is a plant oil (derived from oil seed
a n air compressor set a t 25 psi. Care was taken to ensure rape) and would therefore be a better comparison than
that both the upper and under surfaces of leaves were treated. insecticidal oils derived from petroleum. In this experiment,
When plants had dried, 30 young apterous adult female A. MO was made by adding clarified neem oil (5% by volume) to
pisum (ca. 24-48 h old) were placed in the soil a t the base of MO devoid of neem oil. MO with methanol-extracted neem
the plants. A Mylar cage was placed over the pots to contain oil was made by adding clarified neem oil (5% by volume)
the aphids. The caged plants were arranged in a completely which had been previously extracted with HPLC grade metha-
randomized design on tables in the environmental chamber. nol (sixtimes, 1:lby volume) to MO devoid of neem oil. Canola
Seven days after adult introduction, all aphids were removed oil was added t o MO devoid of neem oil such that the resultant
and counted. This experiment was replicated four times. product contained 5% oil. Canola oil (100%)was formulated
Experiment 2. Effects on First Instars. We also with deionized-distilled water by adding 2.5 mL of oil, 0.2
determined that first instars of A. pisum were the most mL Triton X-100, and 47.3 mL water. MO, MO devoid of neem
susceptible stage to MO (Stark and Rangus, 1994). As such, oil, MO with methanol-extracted neem oil, and MO with canola
we tested the toxicity of MO, Azatin, and RH-9999 to immature oil were compared at the 100 mg of a z a d i r a c h t i f i level as
A. pisum a t the 100 mg of azadirachtidl level. Plants were described in experiment 2 with first instar aphids and clip
treated as in experiment 1,but after drying, one adult aphid cages. Canola oil was tested a t the same level of neem oil
was placed in each of 10 clip cages attached t o the under contained in MO a t the 100 mg of a z a d i r a c h t i f i level.
surface of the leaves (total of 10 aphids per pot). Twenty-four Experiment 7. Identification of the Polar Compo-
hours after introduction of adults, all aphids were removed nents in Neem Oil. Clarified neem oil used in experiment 6
except for one first instar in each cage. This ensured that was analyzed for the presence of chemical constituents.
newly born aphids were exposed a t birth t o the treatments. Clarified neem oil was diluted (1:lO) with 90% methanol and
Plants and aphids were kept in a n environmental chamber at passed through a (2-18 solid phase extraction column (bond
25 "C and 78% RH. Previous data indicated that the adult spec 18, J. T. Baker) and then injected into a n HPLC for
stage was reached in 5 d under our conditions and that MO separation. The HPLC was operated using a 25 cm x 10 mm
a t the 100 mg of a z a d i r a c h t i f i level killed most aphids before C-8 column (Supelco). The elution pattern was operated in a
they reached the adult stage (Stark and Rangus, 1994). This gradient using acetonitrile and water as eluents. The gradient
test was replicated five times and was done to confirm the started a t 28% acetonitrile in water and increased to 95%
results of experiment 1 with the most vulnerable stage of A. acetonitrile in water over 65 min. The flow rate was 4.0 mL/
pisum. min and the UV detector was set a t 215 nm. Fractions from
Experiment 3. Removal of Neem Oil from MO. On the the HPLC were collected and then analyzed by a Finnigan
basis of the results of experiments 1 and 2, we hypothesized mass spectrometer to identify the components. Identifications
that neem oil or a component of the oil, enhances the activity were made by comparing the spectra to an existing data bank
of neem insecticides. This hypothesis was tested in a third as well as proposed structures from the literature. The neem
experiment. MO and MO devoid of neem oil were tested at oil sample was analyzed again after undergoing the methanol
the 100 mg of a z a d i r a c h t i f i level and were compared as extraction process described in experiment 6.
described in experiment 2 with first instars and clip cages. A Data Analysis. Data from all experiments were analyzed
solution of neem oil (5%AI) in water was also prepared. The with one way analyses of variance (ANOVA) and means were
oil solution was tested a t the same concentration of neem oil separated by least significant difference (lsd) (P < 0.05) (SAS
contained in MO at the 100 mg of a z a d i r a c h t i f i level. Institute, 1985) except for the topical tests involving MO
Experiment 4. Addition of Neem Oil to Neem Insec- devoid of neem oil. Here, the data did not meet the criteria of
ticides. This study was conducted to determine whether the normality or equal variances and was therefore analyzed with
addition of neem oil t o azatin and RH-9999 would increase Kruskal-Wallis one-way ANOVA on ranks followed by Stu-
the toxicty of these products. Azatin and RH-9999 were dent-Newmann-Keuls test for mean separation.
compared to Azatin and RH-9999 to which neem oil was added
such that each product contained 5% neem oil prior to dilution
for spray application. This study was conducted in the same RESULTS
manner as experiment 1 with adult aphids and final aphid Experiment 1. Results of experiment 1 indicated
density was the endpoint of interest. This test was replicated that MO, Azatin, and RH-9999 did not affect aphid
five times. population growth in the same manner when the same
Experiment 5. Topical Toxicity. To determine whether
the increase in activity of neem insecticides by the addition of concentration of azadirachtin was applied (Table 1).
neem oil occurs only when the insecticides are applied to plants Aphid population increase was inhibited the greatest
or whether this phenomenon occurs directly in insects we by MO, Azatin was the second most effective product,
conducted topical toxicity studies with MO devoid of neem oil but was much less effective than MO. RH-9999 had
and compared these data to a previously developed data set virtually no effect on aphid population growth.
Neem Oil and Neem Oil Components J. Agric. food Chem., Vol. 43, No. 2, 1995 509

Table 4. Population Density of A. pisum Exposed to Several Neem Insecticides at a Rate of 100 mg of Azadirachtink
(Experiment 4)"
no. of anhids f SD
control Azatin Azatin with neem oil RH-9999 RH-9999 with neem oil
1437.75 f 139.66a 625.25 f 62.09~ 281.75 f 133.79d 1416.0 i 21.34a 810.0 f 101.99b
a ANOVA followed by lsd test. Means followed by the same letter are not significantly different. Based on four replicates.

Table 5. Effects of Margosan-0 and Margosan-0 Devoid Table 6. Effects of Margosan-0 and Margosan-0 Devoid
of Neem Oil on Longevity of Adult A. pisum after Topical of Neem Oil on Reproduction of Adult A. pisum after
Application (Experiment 5) ToDical ADDlication lExDeriment 5)
longevity (days) 5 SEM no. of offspring f SEM
azadirachtin Margosan-0 devoid azadirachtin Margosan-0
concn (mg/L) Margosan-0" of neem oilb concn (mg/L) Margosan-0" devoid of neem oilb
100 10.43 f 1 . 2 4 ~ 16.23 f 0.74b 100 22.87 f 2.86d 75.53 5 3.71b
80 12.00 f 1.20bc 13.50 f 0.95b 80 31.33 5 3.37cd 62.23 f 4.71b
60 12.20 f 1.32bc 15.38 f 0.87b 60 37.03 i 4 . 3 6 ~ 72.03 f 4.00b
40 13.40 f 1.lObc 15.73 i 0.95b 40 46.93 2C 4 . 9 8 ~ 70.43 2C 4.971,
20 15.03 f 1.10b 17.05 f 0.94ab 20 59.07 f 4.24b 72.58 i 5.5713
10 16.00 f 1.00b 15.07 f 1.03b 10 61.70 f 3.6513 67.18 i 5.72b
0 23.10 f 0.98a 18.98 f 0.78a 0 94.37 i 3.73a 91.25 f 3.66a
QThese data are from Stark and Rangus (1994). ANOVA Q T h e ~data
e are from Stark and Rangus (1994). ANOVA
followed by lsd test. Means followed by the same letter are not followed by lsd test. Means followed by the same letter are not
significantly different (P > 0.05). Based on three replicates. significantly different (P > 0.05). Based on three replicates.
Kruskal-Wallis one-way ANOVA on ranks followed by Student- Kruskal-Wallis one way ANOVA on ranks followed by Student-
Newman-Keds test. Means followed by the same letter are not Newman-Keuls test. Means followed by the same letter are not
significantly different (P > 0.05). Based on four replicates. significantly different (P > 0.05). Based on four replicates.

Experiment 2. When MO, Azatin, and RH-9999 devoid of neem oil at the 100 mg of a z a d i r a c h t i d level
were compared against first instar A. pisum, MO was was only 17% lower than controls. It is interesting to
the most toxic product followed by Azatin while RH- note that there was no dose response for MO devoid of
9999 exhibited no toxicity (Table 2). MO was 1.3 times neem oil.
more toxic than Azatin. Experiment 6. The extraction of neem oil with
Experiment 3. When MO, neem oil, and MO devoid methanol resulted in a reduction of activity, but the
of neem oil were compared, MO was again found t o be reduction was not as great as the complete removal of
the most effective product (Table 3). MO was 2.6 times neem oil (MO devoid of neem oil) (Table 7). MO was
more toxic than MO devoid of neem oil. much more toxic than MO devoid of neem oil, which was
Neem oil caused no mortality of A. pisum. However, seen before in experiment 3. MO with methanol-
individuals in the MO devoid of neem oil treatment and extracted neem oil was less toxic than MO, but more
the neem oil treatment were ca. 25% smaller than toxic than MO devoid of neem oil. The addition of
individuals in the controls. canola oil to MO devoid of neem oil resulted in activity
Experiment 4. The addition of neem oil to Azatin similar to MO with methanol-extracted neem oil while
and RH-9999 resulted in a significant increase in the canola oil alone did not cause mortality.
effectiveness of each product (Table 4). Population
growth of A. pisum exposed to Azatin was 43% of the Experiment 7. HPLCMS analysis of neem oil
control while population growth of aphids exposed to indicated that eight major chemicals were present
Azatin with neem oil was only 20% of the control group. (Figure 1). Six of these products, all limonoids, were
Azatin with the addition of 5% neem oil was twice as identified and two remain unknown. The eight products
toxic as Azatin alone. were (A) nimbandiol, (B) deacetylnimbin, (C-1)6-acetyl-
The addition of neem oil to RH-9999 also increased nimbandiol, (C-2) deacetylsalannin, (D) unknown, (E)
its toxicity to A. pisum (Table 4). RH-9999 with 5% nimbin, (F) salannin, and (GI unknown (Chart 1). The
neem oil was 1.75 times more toxic than RH-9999 alone. two unidentified products are believed to be limonoids.
Experiment 5. MO was still more toxic than MO Analysis of neem oil after methanol extraction and the
devoid of neem oil when applied topically (Tables 5 and methanol extract, revealed that all of these chemicals
6). Although longevity was significantly reduced by all were completely removed from the oil (Table 8).
concentrations of MO devoid of neem oil compared to The percent of the eight chemicals in neem oil by
controls, reductions were much lower than in individu- weight and their ratio to azadirachtin in MO and RH-
als exposed to MO. For example, at the equivalent of 9999 is presented in Table 8. Salannin accounted for
100 mg of azadirachtifi, the lifespan of A. pisum the greatest weight of the limonoids found in neem oil.
exposed t o MO-treated broad bean was only 45% of the Nimbin was the second most abundant chemcial fol-
control group while the lifespan of A. pisum exposed to lowed by deacetylnimbin, unknown D, and unknown G.
broad bean treated with MO devoid of neem oil was 85% In MO, salannin was also the major limonoid, unknown
of the control (Table 5). Production of offspring was also D was second, and nimbandiol was third. Limonoids
was significantly reduced by all concentrations of MO could not be detected in neem oil that had been
devoid of neem oil when compared to controls (Table 6). extracted with methanol. The ratio of limonoids in RH-
However, the reduction in offspring was much more 9999 was different than that found in MO. For ex-
pronounced in individuals exposed t o MO. Individuals ample, five times more deacetylnimbin was present in
exposed to MO a t the 100 mg of azadirachtifi level, RH-9999 than in MO and five times more nimbandiol
produced 76% fewer offspring compared to controls. was present in MO than in RH-9999. Salannin levels
Offspring production by individuals exposed to MO were about the same in both products.
510 J. Agric. Food Chem., Vol. 43, No. 2, 1995 Stark and Walter
Table 7. Toxicity of Neem Insecticides to A. pisum Exposed as First Instars to Margosan-0,Margosan-0 Devoid of Neem
Oil, Margosan-0 with Methanol-ExtractedNeem Oil (5%), and Margosan-0 with Canola Oil (5%)All at the Equivalent of
100 mg of Azadirachtia and Canola Oil (6%) (Experiment f3)=
% mortalitv f SD
Margosan-0 Margosan-0 with Margosan-0
control Margosan-0 devoid of neem oil methanol-extracted neem oil with canola oil canola oil
2.5 f 5.0d 97.7 i 5.0a 37.5 f 5 . 0 ~ 70.0 k 8.16b 65.0 i 12.9b 2.5 f 5.0d
a ANOVA followed by lsd test. Means followed by the same letter are not significantly different. Based on four replicates.

R=Ac 6.acelylnimbandiol cz n
deacelylsalannin

A /
R=H nimbandiol

I I I I ( I l I I 1 1 1 1 ( 1 1 1 1 ) 1 1 1 1 ( 1 1 1 1
2'0 40
T i m e [min]
Figure 1. 1. HPLC chromatogram of neem oil prior t o extraction with methanol.

DISCUSSION different between RH-9999 and MO which might influ-

Results of this study clearly show that neem insecti- ence efficacy.
cides are not equal in terms of pea aphid control. When The removal of neem oil from MO greatly reduced its
applied a t equivalent rates of azadirachtin, MO was toxicity while adding neem oil to Azatin and RH-9999
significantly more toxic than the other neem insecti- resulted in increased toxicity. Thus, neem oil andor a
cides. Azatin was the second most toxic product while component of the oil influenced insecticidal activity of
RH-9999 was virtually nontoxic. The reason that RH- azadirachtin, the active ingredient in these neem in-
9999 was not toxic to A. pisum may have t o do with the secticides.
fact that azadirachtin is hydroxylated in this product.
Hydroxylated azadirachtin may not penetrate plant When we topically applied MO and MO devoid of
tissue or insect cuticle as readily as unhydroxylated neem oil, we found that the lack of oil greatly reduced
azadirachtin. Other reasons for differences in toxicity the efficacy of MO. By eliminating the host plant in
might have to do with formulation and the content of this experiment we showed that the phenomenon we
other limonoids. RH-9999 is a wettable powder while observed with neem oil also occurred when aphids were
MO and Azatin, the more effective products, are emul- directly exposed. Thus, we showed that the increase
sifiable concentrates. Also, the ratios of limonoids were in activity of neem insecticides by the addition of neem
Table 8. Weight of Liminoids and Their Ratio to Azadirachtin in Various Neem Materials
wt % of limonoids (ratio limonoids/azadirachtin)
neem material azadirachtin nimbandiol deacetylnimbin 6-acetylnimbandiol unknown d nimbin salannin unknown G
neem oil 0.01 0.27 (27) 1.7 (170) 0.06 (6) 1.6 (160) 2.2 (220) 4.5(450) 1.3(130)
neem oil extracted NDa ND ND ND ND ND ND ND
with methanol
Margosan-O 0.25 2.4 (9.6) 1.4 (5.6) 1.1(4.4) 6.3 (25.2) l ( 4 ) 7.3 (29.2) 0.2 (0.8)
RH-9999 20.0 0.48 (0.02) 7.0 (0.35) 0.3 (0.01) 4.2 (0.21) 3.2 (0.16) 7.4 (0.37) 0.7 (0.04)
ND = ~ 0 . 0 1 %wt 96.
Neem Oil and Neem Oil Comoonents J. Agric. Food Chem., Vol. 43, No. 2, 1995 511
Chart 1 There may be other factors responsible for the in-
creases in efficacy of insecticides in the presence of oils
such as increased persistence on crops (Salt and Ford,
1984; Hesler and Plapp, 1986).
Xie and Isman (in press) found that tall oil, a product
of the pulp wood industry, enhanced the activity of
azadirachtin in the variegated cutworm, Peridroma
saucia. These authors concluded that tall oil worked
A R=H niinbniidiol C2 Deace!ylsnlannin by increasing cuticular penetration of azadirachtin.
In our study, the substitution of canola oil for neem
oil in MO resulted in a 33% loss of toxicity. The
substitution of neem oil extracted with methanol re-
sulted in a 30% loss of toxicity, but complete removal
of oil from MO resulted in a 62% loss in toxicity. So
the addition of oils increased the toxicity of MO, but
COiR CO2Me something that was removed from the oil during the
methanol extraction process resulted in a 30% loss in
B R=R'=Me deacetylnimbin E Niinbin toxicity. Neem oil alone did not kill A. pisum within
seven days after exposure, but individuals were much
smaller than controls indicating biological activity of the
oil.
Therefore, two processes are obviously at work. Oils
increase toxicity, probably by increasing penetration,
and a polar component(s)in neem oil also increases the
toxicity of MO.
Mass spectral evaluations of neem oil before and after
C1 R=Ac 6=acetylnimbandiol F Salannin methanol extraction revealed the presence of six li-
monoids and two unknown compounds. We hypothesize
oil occurs both directly in insects as well as when the that one or more of these products is responsible for 30%
insecticide is applied t o plants. of the toxicity of MO. The importance of this finding
Some oils are known to increase insecticidal activity cannot be determined until further work is done.
(Sun, 1968). It is generally believed that the increase For future studies the following questions should be
in efficacy caused by oils is mediated by increasing addressed:
penetration into crops and insects and by increasing 1. Does the observed enhancement by neem oil occur
coverage and persistance (Anderson et al., 1986; de only in the pea aphid or is it manifested in other aphid
Licastro et al., 1983; Treacy et al., 1986, 1991). There- species and other pest and beneficial species?
fore, the increase in toxicity caused by oils is not 2. Does neem oil or canola oil actually increase
considered synergism, which is a situation where the penetration of azadirachtin or is some other mechanism
effect of two compounds with similar modes of action, involved? Toxicodynamic studies with MO and MO
applied together or consecutively, is greater than would devoid of neem oil should be conducted to prove whether
be expected from the sum of the individual effects (Bliss, greater penetration of azadirachtin into plants and
1939). Another term, potentiation, is used to describe insects is occurring.
a situation where synergism occurs, but the two com- 3. What component(s)of neem oil is (are) responsible
pounds have different modes of action. Sun and Johnson for the enhanced activity we observed?
(1972) used the term quasisynergism for cases of Results of this study may have implications for the
increased cuticular penetration in insects and went on development and use of neem insecticides in the future.
to define "total synergism" as the product of enzymatic Neem oil is phytotoxic to some crops and thus the
inhibiton and synergism due t o an increase in penetra- decision to eliminate it from a commercial insecticide
tion. may be very prudent depending upon the crops that are
An example of an oil increasing the efficacy of an targeted for registration. However, our results show
insecticide was shown by Sun (1968). He found that that addition of neem oil to neem insecticides increases
carbaryl was much more toxic to the house fly, Musca their efficiacy at least with the pea aphid. If the same
domestica L., when it was applied with kerosene than thing occurs with other pest species, addition of neem
with acetone. Anderson et al. (1986) showed that sun oil t o future neem insecticide formulations may result
spray oil acted as a potentiator of avermectin in in better control.
Spodoptera eridania (Cramer), but not in Heliothis
virescens (F.) after these insects ingested insecticide- ACKNOWLEDGMENT
treated Sieva bean, Phaseolus lunatus L., foliage. We thank Theresa Rangus for her assistance with this
Mineral oil synergized the toxicity of cyfluthrin study, Cliff Hull for HPLC analysis and quantification
compared to water in field tests with the boll weevil, of limonoids, and Abdul Mabud for NMR identification
Anthonomus grandis grandis Boheman, feeding on of the limonoids. We also thank Agridyne Technologies
insecticide-treated cotton (Treacy et al., 1991). Paraffin- Inc., Salt Lake City, UT, and Rohm and Haas Co.,
ic oils increased penetration of insecticides in the Philadelphia, PA, for supplying Azatin and RH-9999,
conenose bug, Triatoma infestants (de Licastro et al., respectively.
1983). However, a study by Southwick et al. (1983)
showed just the opposite in cotton. The addition of LITERATURE CITED
cotton seed oil and soy bean oil to permethrin actually Anderson, T. E.; Babu, J. R.; Dybas, R. A.; Metha, H.
slowed penetration into cotton leaves compared to an Avermectin B1: Ingestion and contact toxicity against
aqueous formulation. Spodoptera eridania and Heliothis virescens (Lepidoptera:
512 J. Agric. Food Chem., Vol. 43, No. 2, 1995 Stark and Walter

Noctuidae) and potentiation by oil and piperonyl butoxide. Nilaparvata lugens CHomoptera: Delphacidae). J . Econ.
J . Econ. Entomol. 1986,79,197-201. Entomol. 1984,77,502-507.
Ascher, K. R. S. Nonconventional insecticidal effects of pesti- Schmutterer, H. Properties and potential of natural pesticides
cides available from the neem tree, Azadirachta indica. from the neem tree, Azadirachta indica. Annu. Rev. Ento-
Arch. Insect Biochem. Physiol. 1993,22,433-449. mol. 1990,35,271-297.
Bliss, C. I. The toxicity of poisons applied jointly. Ann. Appl. Southwick, L. M.; Clower, J. P.; Clower, D. F.; Graves, J. B.;
Biol. 1939,26,585-615. Willis, G. H. Effects of ultra-low-volume and emulsifiable-
de Licastro, S. A.; Zerba, E. N.; Casabe, N. The relation concentrate formulations on permethrin coverage and per-
between viscosity and penetration of some diethyl p-sub- sistence on cotton leaves. J . Econ. Entomol. 1983,76,1442-
stituted phenyl phosphorothionates and oil carriers into the 1447.
cuticle of Triatoma infestants. Pestic. Biochem. Physiol. Stark, J . D. Comparison of the impact of a neem seed kernel
1983,19,53-59. extract formulation, Margosan-0 and chlorpyrifos on non-
Hesler, L. S.;Plapp, F. W., Jr. Combinations of oils and similar target invertebrates inhabiting turf grass. Pestic. Sci. 1992,
compounds with insecticides: effect on toxicity and on leaf 36,293-299.
residues. Southwest. Entomol. Suppl. 1986, 11, 75-82.
Hull, C. J.; Dutton, W. R.; Switzer, B. S. Quantitation of Stark, J. D.; Rangus, T. M. Lethal and sublethal effects of the
azadirachtins in insecticidal formulations by high perfor- neem insecticide formulation, Margosan-0, on the pea
mance liquid chromatography. J . Chromatogr. 1993, 633, aphid. Pestic Sci. 1994,41,155-160.
300-304. Stark, J. D.; Wong, T. T. Y.; Vargas, R. I.; Thalman, R. K.
Kale, R. D.; Bano, K.; Vinayak, K.; Bagvaraj, D. J. Suitability Survival, longevity, and reproduction of Tephritid parasi-
of neem cake as a n additive in earthworm feed and its toids (Hymenoptera: Braconidae) reared from fruit flies
influence on the establishment of microflora. J . Soil Biol. exposed t o azadirachtin. J. Econ. Entomol. 1992,85,1125-
Ecol. 1986,6,98-103. 1129.
Koul, 0.; Isman, M. B.; Ketkar, C. M. Properties and uses of Sun, Y-P. Dynamics of insect toxicology-A mathematical and
neem-Azadirachta indica A. Juss. Can. J . Bot. 1990, 68, graphical evaluation of the relationship between insect
1-11. toxicity and rates of penetration and detoxication of insec-
Mansour, F.;Ascher, K. R. S.; Omari, N. Effect of neem seed ticides. J . Econ. Entomol. 1968,61,949-955.
kernel extracts from different solvents on the predacious Sun, Y-P.; Johnson, E. R. Quasi-synergism and penetration
mite Phytoseiulus persimilis and the phytophagous mite of insecticides. J . Econ. Entomol. 1972,65,349-353.
Tetranychus cinnabarinus as well as on the predatory spider Treacy, M. F.; Benedict, J. H.; Schmidt, K. M. Toxicity of
Chiracanthium mildei. In Natural pesticides from the neem insecticide residues to the boll weevil: comparison of ultra-
tree and other tropical plants, Proceedings of the 3rd low volume/oil vs. conventionaVwater and water-oil sprays.
International Neem Conference, Nairobi, 1986; Schmut- Southwest. Entomol. Suppl. 1986,11, 19-24.
terer, H., Ascher, K. R. S., Eds.; German Agency for Treacy, M. F.; Benedict, J. H.; Schmidt, K. M.; Anderson, R.
Technical Cooperation (GTZ): Eschborn, 1987; pp 577-587. M. Mineral oil: Enhancement of field efficacy of a pyrethroid
Rossner, J.; Zebitz, C. P. W. Effects of soil treatment with neem insecticide against the boll weevil (Coleoptera: Curculion-
products on earthworms (Lumbricidae). In Natural pesti- idae). J. Econ. Entomol. 1991,84, 659-663.
cides from the neem tree and other tropical plants, Proceed- Xie, Y. S.; Isman, M. B. Tall oil: enhancement of neem and
ings of the 3rd International Neem Conference, Nairobi, azadirachtin toxicity to the variegated cutworm, Peridroma
1986; Schmutterer, H., Ascher, K. R. S., Eds.; German saucia. J . Appl. Entomol. 1994,in press.
Agency for Technical Cooperation (GTZ): Eschborn, 1987;
pp 611-621.
Salt, D. W.; Ford, M. G. The kinetics of insecticide action. Part
Received for review June 28, 1994. Accepted November 22,
111: the use of stochastic modeling to investigate pick-up of 1994.@
insecticides from ULV-treated surfaces by larvae of
Spodoptera littoralis Boisd. Pestic. Sci. 1984,15,382-410. JF940342M
SAS Institute. SAS user’s guide: statistics; SAS Institute:
Cary, NC, 1985.
Saxena, R. C.; Justo, H. D., Jr.; Epino, P. B. Evaluation and @ Abstract published in Advance ACS Abstracts, Janu-
utilization of neem cake against the brown planthopper, ary 1, 1995.