TOXICOLOGICAL STUDIES ON BORIC ACID,

IMIDACLOPRID AND FIPRONIL AND THEIR BINARY

MIXTURES AS INSECTICIDES ON GERMAN COCKROACH
Blattellagermanica (L.) (DICTYOPTERA: BLATTELLIDAE)

By

FATMA SHERIF AHMED

B.Sc. Agric. Sci. (Pesticides), Fac. Agric., Cairo Univ., 2007

THESIS
Submitted in Partial Fulfillment of the
Requirements for the Degree of

MASTER OF SCIENCE

In

Agricultural Sciences
(Pesticides)

Department of Economic Entomology and Pesticides

Faculty of Agriculture
Cairo University
EGYPT

2015

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INTRODUCTION
The German cockroaches, Blattellagermanica (L.), (Dictyoptera,
Blattellidae) are the most common indoor pests, especially in multiple-family
housing and the most significant pest in many parts of the world (Goddard,
2003). German cockroaches prefer warm, wet locations with high humidity such
as kitchens, bathrooms and laundry areas. These conditions are available in
several places as homes, apartments, restaurants, supermarkets, hospitals and
other buildings where food are stored.
Cockroaches are not only corrupt food but also transfer pathogens such as
Salmonella, Shigella, Escherichia coli, Staphylococcus aureus and Bacillus
cereus (Baumholtz et al., 1997 and Tachbeleet al., 2006). Medically important
parasites such as bacteria, fungi and molds, protozoans, viruses were isolated
from external and internal surface of cockroach (Brenner, 1995). Cockroaches
can also transfer both gram-positive and negative bacteria (El-Sherbini and El-
Sherbini, 2011).
A large number of neurotoxic and non-neurotoxic insecticides were used
for German cockroach control, as this pest has a considerable ability to develop
resistance to a variety of chemical insecticides(Cochran, 1989 and 1995a; Scott
et al., 1990; Rust and Reierson, 1991; Rust et al., 1993; Holbrook et al., 1999;
Espinosa-Islas et al., 2002 and Rahayuet al., 2012). The toxicity of some new
neurotoxic insecticides is either very limited (i.e., neonicotinoids) or not well
established (i.e.,phenylpyrazole) against German cockroach in Egypt.
Boric acid (H3BO3) has been used as an insecticide since 1948 (See et al.,
2010). It was the standard chemical used for the control of some pests until the
chlorinated hydrocarbon pesticides become available (Lal and Srivastava,
1950). Moreover, it is not classified as carcinogen or skin irritant (Langford et
al., 1954). One advantage of using boric acid is that, it destroys the cellular
lining of the foregut. This effect induces death to the insects by starvation and

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the cockroaches have difficult to show any resistance to boric acid (Cochran,
1995b).
A phenylpyrazole insecticide fipronil discovered by Bayer Crop Science
in 1987, and introduced in 1993 as a form of solid (e.g., insect bait), liquid
spray, or as a granular product (Anonymous, 2000), and registered in the U.S. in
1996 (Ware, 2000 and Jeong Lee et al., 2010). It controls cockroaches,
mosquitoes, ticks, and fleas at both larval and adult stages (Chantonet al.,
2002). Cole et al. (1993) found that fipronil interferes with the δ-aminobutyric
acid (GABA) channel, where it disrupts normal nerve influx transmission (e.g.,
passage of chloride ions) this causes excessive neural excitation, paralysis, and
finally insect death (Gant et al., 1998). Fipronil gel baits have been widely used
to control German cockroach in America since several years due to german
cockroach resistance to several insecticides and fipronil gel bait consider more
safe in application than other treatment methods (Ahmed and Suliyat, 2011).
A neonicotinoid insecticide, imidacloprid belonging to the
chloronicotinyl compounds. It was first proposed in 1991. It exerts toxic effects
after oral ingestion; it is also suitable for use in bait formulations (Nasirian,
2007). The main factor of chloronicotinyl being favorable is their low affinity to
vertebrate compared to insect nicotinic receptors. They have high selectivity;
have a quick knock down effect and long residual activity. Worldwide, it is
considered to be one of the insecticides used in the largest volume (Cox, 2001).
Nasirian (2008) reported that fipronil and imidacloprid gel baits completely
killed the German cockroaches under laboratory conditions in ingested bait
method.
Poisoned baits, in the last two decades, have largely displaced other
formulations for controlling of German cockroaches and other pests (Anaclerio
and Molinari, 2012). The poisoned baits are much less translocatable, so it is
less hazardous than the spray formulations (Gore and Schal, 2004). So that the
tested insecticides used as a bait to be safer and environmentally friendly than

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insecticide sprays. The toxicity of fipronil baits has been estimated from
mortality rates (Scott, 1991 and Ross, 1993) and consumption rate (Wang et al.,
2006).
The three tested compounds (boric acid, imidacloprid and fipronil) were
selected on the basis of their high efficacy against German cockroach. Although
fipronil hasn't been used in Egypt against German cockroach yet, it widely used
in all developed countries as a highly effective tool for controlling the German
cockroach especially in bait form. The present study evaluated the efficacy of
fipronil, imidacloprid and boric acid on the nymphal instars and adult males of
German cockroach. On the other hand, German cockroach didn't appear to be
resistant to boric acid compound although this compound is used against this
insect for ages. Also, the application of neonicotinoid insecticide imidacloprid
in bait form is easy to apply in out-of- way places and is stable under a wide
range of conditions.
The objectives of current study are:
1. Determination of the oral toxicity of boric acid, imidacloprid and
fipronil as poisoned baits against different stages of B. germanica (L.).
2. Evaluation of the palatability and the toxicity of fipronil incorporated
into different food baits on the 6th instar nymphs of German cockroach
B. germanica (L.).
3. Comparing the toxicity of imidacloprid as topical application and
stomach poisoned bait against the different stages of the tested insect.
4. Estimation of the lethal times (LT50) of the two tested compounds on
the different stages.
5. Evaluation of the effect of binary mixtures of the three tested
insecticides on 6th instar nymphs.

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MATERIALS AND METHODS

Tested insect (Rearing and Manipulation the Insect):

German cockroach, BlattellaGermanica (L.)
Family: Blattellidae
Order: Dictyoptera
Cockroaches are insects with three stages: eggs, nymphs, and adults,
males and females. About one week after mating, the female cockroach deposits
eggs into a protective covering called the oetheca or egg case which protects the
eggs from drying. The egg case looks like a tiny, brown, purse-shaped capsule,
it is about 8 mm long, 3 mm high, and 2 mm wide (Tanaka, 1976). The eggs
hatch into a small but fully-developed nymphs that grow through a series of
molts, shedding its old cuticle (exoskeleton) at each molt. The range of head
capsule widths in the lst to 6th instars nymphs are, respectively: 0.74-0.85 mm;
0.88-1.05 mm; 1.02-1.35 mm; 1.20-1.62 mm; 1.40-1.86 mm; and 1.84-2.25 mm
(Murray, 1967). Nymphal survival percentages from egg hatch to the adult stage
was estimated at 85- 87% (Willis et al., 1958 and Archboldet al., 1987).
The nymphs of cockroaches emerge as fully-winged adults after the final
sixth molt. Newly emerged adults appear white, but become fully pigmented
within 24 hours (Ross and Mullins, 1995). As the nymphs develop faster, they
gain more body mass and the adults are heavier, the females produce more
fertile oethecae (Gemenoet al., 2001).
German cockroaches are semi-social insects; all their stages and ages are
found at the same place. Colony of German cockroach BlattellaGermanica (L.)
used for the present study was obtained from Aburwash, Giza Governorate,
reared for two years and the all bioassay tests were carried out in the
Department of Economic Entomology and Pesticides, Faculty of Agriculture,
Cairo University, Egypt. The average number of (eggs per capsule, capsules per
female), period between egg case formation to hatching, number of molts,

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nymphal stage (in days), life cycle and average longevity of adults (in days) are
given in the following.

Cockroaches insects were reared in plastic containers (1 liter capacity)

and maintained at 27 ± 1°C, 60±5% relative humidity in an incubator at a
photoperiod of 12:12 dark-light. A wet piece of cotton was placed in each jar as
a water source for the cockroaches with some of modifications as described by
(Nalyanya and Schal, 2001; Nasirianet al., 2006a and 2011; Sulaimanet al.,
2007 and Kilani-Morakchiet al., 2009). The cockroaches for rearing were fed on
a source of protein and a sugar solution coated on a piece of cotton, and a
cardboard or a plastic tubes as a shelter (Nasirianet al., 2006a,c).
The present investigations were carried out on the 2nd, 6th instar nymphs
(two days old) and adult males (2-3weeks old). Only adult males, not adult
females, were used in the experiments because their weight and physiology are
more uniform than adult female (Appelet al., 1983). The females were also
needed for further reproduction.
Insecticides tested:
Three commercial formulations were used in the toxicological study.
These compounds are representing boric acid (inorganic compound),
imidacloprid(neonicotinoid insecticide group) and fipronil (phenylpyrazole
group). The common name, trade name and properties of the tested compounds
(boric acid, imidacloprid and fipronil) are given in Table 2.
a. Boric acid insecticide:
Boric acid (H3BO3) was used as an insecticide for control of cockroaches,
termites, fire ants, fleas, silverfish, and many other insects(See et al., 2010). It is
considered to be safe to use in household kitchens to control cockroaches and
ants.The dry powder is abrasive to the insect and may also have some toxic
effects on the nervous system of insects (Habeset al., 2005). It acts as a stomach
poison (affecting the insect’s metabolism) and as a contact poison.

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b. Imidacloprid insecticide:
Imidacloprid is an insecticide belonging to the neonicotinoid class of
compounds. It was first proposed in 1991 and registered for use by the United
States Environmental Protection Agency (U.S. EPA) in 1994. It is used to
control a wide variety of agricultural, urban, public health, and veterinary pests
and is estimated to account for 11-15% of the total global insecticide market
(Tomizawa and Casida, 2005).
c. Fipronil insecticide:
Fipronil is a phenylpyrazole insecticide discovered by Bayer Crop
Science Company), and introduced as form of solid (e.g., insect bait), liquid
spray, or as a granular product (Anonymous, 2000). It was registered in the U.S.
in 1996 (Ware, 2000; Connelly, 2001 and Jeong Lee et al., 2010). It was made
available in 1999 in bait formulations for use against the German cockroach,
BlattellaGermanica(L.).
Tested application treatments:
a. Toxicity studies of fipronil, imidacloprid and boric acid against the
6thnymphal stage:
The three tested compounds fipronil, imidacloprid and boric acid were
tested to know their toxicity on the last 6thnymphal stage of German cockroach,
B. Germanica. Each compound was incorporated into the diet at different
gradual concentrations. Experimental baits with 0.1% sodium benzoate to
prevent fungal growth were placed in each glass jar (in control diet and treated
diet jars). All tested insecticides used as bait were prepared in water solution
that it is important to note that cockroaches drink regulatory and survive longer
in higher humidity than in lower humidity percentage (Cochran, 1983 and
Dambach&Goehlen, 1999).

Cockroaches were not fed before any experiments for 3 days in order to
increase their hungry level and optimize their feeding on the toxic baits

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(Durierand Rivault, 2000). According to Young et al. (1999) nymphs consumed
a large ration of food after being starved, then it return to the normal feeding
pattern, and quickly resume normal development.
Boric acid, imidacloprid and fipronil were separately incorporated with
diet and orally administrated to the tested cockroaches. Eight different gradual
concentrations of boric acid (1.25, 3.5, 7.0, 12.5, 19.0, 25.0, 37.5 and 50%)
were tested. Also imidacloprid was incorporated into diet and orally
administered at different diluted seven concentrations (1250, 2500, 5000,
10000, 15000, 20000 and 28000 ppm a.i.). Fipronil was also incorporated into
the diet at 9 serial diluted concentrations 1.5, 3.0, 6.0, 12.5, 25.0, 50.0, 75.0,
100 and 125 ppm a.i.
Each concentration of boric acid, imidacloprid and fipronil was applied
separately to 30 individuals of the last nymphal (6 th instar nymph) stage of B.
Germanica (L.) divided into three replications (10 individuals/ replicate). The
tested nymphs were exposed to the poisoned baits for 7 days in case of boric
acid and imidacloprid, but for fipronil the exposure period to the poisoned bait
was only 3 days.
The check cockroaches nymphs (control group) provided with untreated
diets (beef+ powdered milk+ sugar solution), coated on a piece of cotton and
cardboard on a plastic tubes as a shelter (Nasirianet al., 2006c). Thirty
untreated 6th instar nymphs (control groups) were divided into 3 replicates (10
individuals/ replicate) and provided with untreated food and lifted for 3 or 7
days, after these periods the dead nymphs were recorded. The mortality
percentages in the treated nymphs groups (i.e.fipronil, imidacloprid and boric
acid) were also recorded and corrected for the natural mortalities by using the
Abbott’s formula (Abbott, 1925) and calculated to obtain the Net % kill as
following formula:

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 X-Y
Net % kill = ---------------- × 100
100- Y

Where: X= Mortality percentage in treatment.

Y= Mortality percentage in control group
The LC50 and LC90 values were calculated.
b. Palatability of tested baits treated with fipronil on 6thnymphal stage:
In this study, four laboratory prepared baits were used. These baits
contain different components and administrated to the cockroaches to choice the
most palatable and toxic baits depending on their consumption rate and
mortality percentages. Such baits were:
a. Beef bait: (beef, powdered milk and sugar 2:1:1, weight/ weight).
b. Onion bait: (onion, sugar, flour and industrial margarine 2:1:1:2, w/w).
c. Strawberry gel bait: (strawberry gel, sugar and flour 2:1:1, w/w).
d. Sweet honey bait: (sweet honey, rusk and flour 2:1:1, w/w).

The palatability of the four tested different baits (i.e. beef, onion,
strawberry gel or sweet honey bait) treated with fipronil was estimated by
calculating consumption rate and mortality percentage. The consumption rate
was conducted by weighing the tested bait before and after treatment. Fipronil
was selected on the base of being highly effective insecticide against German
cockroach depending on the bioassay tests in the present study. The mortality
was conducted from adding the LC50 and LC90 value of fipronil, 22.5 and 107.5
ppm, respectively, as calculated by the Probit toxicity line.
Thirty insects of last nymphal stage or adult males of B. Germanica were
transferred to each treatment and divided into three replicates (10 individuals for
each) for the different concentrations of fipronil were conducted.
The control insects were divided into two groups, the first one had each
type of the tested diets with cockroaches, and the second control group had the
diet without cockroaches for estimation of water loss to be excluded from the
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consumption rate (Wang et al., 2006). Consumption rate of bait was calculated
by the following:

(W1- W2) - WC2

Consumption rate % = --------------------- X 100
Wc1

Where, W1 and W2 are the weight of bait treatment groups before and
after exposure to cockroaches, respectively. Wc1 is the weight of bait in control
groups with untreated bait, whereas Wc2 is the weight of tested diets without
cockroaches for estimation of water loss.

c. Toxicity of the three tested insecticides on different nymphal stage and

adult males:

As mentioned before the selected individuals, nymphs and adult males

were left to habituate for 3 days before the treatments. Fipronil was evaluated
on 3 different instars of nymphal stage (2nd, 4th and 6th instar) or adult males of
the German cockroach. The methods of treatments were the incorporation into
diet technique. Fipronil was prepared at 9 serial concentrations (1.5- 125 ppm)
against 2nd, 4th and 6th instar nymphal stage and adult males. Thirty individuals
were divided into three replicates, 10 cockroaches for each. Control groups
received only untreated diet. The mortality percentages were recorded after 24,
and 72 hours. The LC50 and LC90 were conducted after 24 and 72 h.
Imidacloprid and boric acid compounds were tested against different
stages, 2nd and 6th instar nymphs and adult males of B. Germanica. Thirty
individuals of cockroaches were transferred to each treatment in 3 replicates.
Each replicate of 10 cockroaches was hold in a glass jar covered with muslin
and rubber band.
Boric acid was incorporated only as a stomach poison into the diet and
orally administrated at eight different concentrations from 1.25 to 50.0%.
١٠
 Imidacloprid was evaluated on three different stages of the German
cockroach (2nd and 6th instar nymphs and adult males), using two techniques 1)
orally treatment by incorporating into the treated food baits, and 2) obligatory
contact with treated glass-jars. Thirty of 2nd and 6th instar nymphs or adult males
of B. Germanica were selected to each treatment, divided into three replicates
(10 individuals for each) of the different concentrations.
For application of imidacloprid insecticide as a stomach poison, it was
incorporated into the diet and orally administrated at 7 different diluted
concentrations (2500- 28000 ppm) against 2nd and last nymphal stage and adult
males of the German cockroach. The insects were exposed to the insecticide for
7 days, after that the insects was allowed to eat untreated food for other 7 days.
The LC50 and LC90 were estimated after 7 and 14 days, respectively.
For application of imidacloprid insecticide as a contact poison, the inside
surface of jars (1 liter capacity) was covered with imidacloprid by residual film
technique. Different concentrations of imidacloprid were prepared by dilution
with acetone (10 ml for each replicate), the treated jars were rolled horizontally
until all of the acetone had evaporated, so that the insecticide was deposited
evenly over the inner surface of the jar (Nasirianet al., 2006c). The insects were
exposed to the treated jars with 7 different diluted concentrations (1250- 28000
ppm). The insects were exposed to the treated jars for 7 days, then the survival
individuals were moved to clean jars and exposed to untreated diet for other 7
days. The LC50, LC90 were estimated after 7 and 14 days, respectively.

Determination of the lethal time concentrations (LT50 and LT90 values):

The LT50 and LT95 values were estimated every two days for the different
concentrations after treatments along two weeks (i.e. 3, 5, 7, 9, 11 and 14 days)
and recoded to measure the effect of the period of exposure (lethal time
concentrations) in the tests of boric acid and imidacloprid insecticides against
the 2nd and 6th instars nymphs and adult males of B. Germanica (L.). The insects

١١
were exposed to the treated baits for 7 days, after that the survival individuals
were transferred to clean jars and exposed to untreated diet. The mortality
responses were recorded after 7 and 14 days and the LC50, LC90 were estimated.
Control groups were fed only on untreated diet.

Effect of binary mixture of the three different tested insecticides against the
6thnymphal stage of German cockroach:

In order to assess the joint action of the tested compounds (fipronil,

imidacloprid and boric acid), different binary combinations of these compounds
at their LC25 values were tested on the 6th instar nymphs. The binary mixtures
were prepared before treatment at ratios of 1: 2, 1: 1 and 2: 1. For each
treatment, there were 30 insects in three replicates (10 individuals for each)
were used. The mortality percent of each mixture were recorded after 6 days as
observed mortality.
The combined action of the different mixtures was expressed as the co-
toxicity factor which was estimated according to the equation given by Mansour
et al., (1966).
Observed mortality – Expected mortality
Co- Toxicity factor = ------------------------------------------------ X100
Expected mortality

Generally, the co- toxicity factor of value 20 or above indicates

potentiation effect, while the negative co-toxicity values from -20 or less
indicates antagonism (inhibition) effect. The co- toxicity factor values between
> -20 and < 20 indicates an additive effect.
Statistical analysis of the obtained data
The mortalities of tested insects were recorded for fipronil after 24 and 72
h, but for boric acid and imidacloprid after 7 and 14 days. All mortalities were

١٢
corrected using Abbott’s formula (Abbott, 1925). The LC50 and LC90 were
estimated after treatments of tested compounds. Also, consumption rate of the
untreated and poisoned baits was calculated by the formula of (Wang et al.,
2006).
The lethal time (LT50 and LT90) were also estimated for the different
concentrations of the tested insecticides, boric acid and imidacloprid and
recoded after 14 days. The LC50 was estimated every two days after the
treatment of boric acid and imidacloprid through two weeks after treatments and
plotted to measure the significance of the increased period of exposure.
The combined action of the different binary mixtures was expressed as
the co-toxicity factor which was estimated according to the equation given by
Mansour et al. (1966). Results are expressed as means ± standard error (SE).
The significance between control and treated series was estimated using t-test at
5% level.
The toxicity lines were plotted according to Finney analysis (Finny,
1971). Results from all treatments were tested with χ2 (calculated Chi – Square
test) for Heterogeneity. The software program Micro origin was used for
plotting the histograms.

RESULTS

1. Evaluation of different poisoned baits on the last nymphal stage (6th) of

german cockroach B. germanica

a. Determination of the toxicity (Bioassay tests) of fipronil, imidacloprid

and boric acid against the last nymphal stage of B. germanica(L.)

According to the LC50 and LC90 values, it could be generally concluded

that fipronil proved to be highly toxicagainst German cockroaches, which gave
the lowest LC50 and LC90 (22.5 and 107.5 ppm, respectively) followed

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ascending by imidacloprid (9228 and 24980 ppm, respectively) and then boric
acid (9.87% and 46.3 %, respectively).

b. The food consumption rates and mortality percentages of different

components baits:
Four different baits contains different food constituents were tested for
evaluate their attractancy and mortality percentages resulted by adding either
the LC50 or LC90 of fipronil. At the LC90 level, the beef bait with fipronil caused
the highest rate of consumption (97.3%) and the highest mortality percentage
(93.3%). According to the calculated mortality percentage, the baits
effectiveness at the LC90 level of fipronil could be arranged in a descending
order as follow: beef bait (93.3%) ˃ onion bait (80.0%)> strawberry gel bait
(60.0%)> sweet honey bait (46.7%). Finally, it could be concluded that the beef
bait had the highest consumption rate and mortality%. So beef poisoned bait
could be recommended to control german cockroaches especially in high
infested area.

2. Toxicity of fipronil as a poisoned bait against different nymphal instars

(2nd,4th and 6th) and adult males of german cockroach B. germanica after
24 and 72 hours of exposure.

For 2nd instar nymph, the LC50 and LC90 values were 9.5 and 44.9 ppm
after 24 hours, these values were 2.3 and 24.7 ppm after 72 hours, respectively.
For 4th instar nymphs, the LC50 and LC90 values after 24 hours were 15.9 and
69.8 ppm and they were 6.5 and 56.7 ppm after 72 hours, respectively. For the
last 6th instar nymph, the LC50 and LC90 were 22.5 and 107.5 ppm after 24 hours
and these were 12.5 and 53.9 ppm after 72 hours, respectively.
The LC50 and LC90 of fipronil against German cockroach adult males
were 38.9 and 221.5 ppm after 24 hours, and they were 14.2 and 107.6 ppm
after72 hours, respectively.

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 The median lethal time LT50 of fipronil against 2nd instar nymph was 37.4,
12.9 and 6.6 hours for the concentrations of 6, 12.5 and 25 ppm, respectively.
For the 4th instar nymphs, The LT50 value of concentrations 12.5, 25 and 50 ppm
were 42.9, 16.7 and 8.4, 33.9 h, respectively. For the 6th instar nymphs, The
LT50 value of concentrations 25, 50, 75 and 100 was 22.7, 15.05, 13.02, and
6.63 h, respectively.
For adult males, The LT50 values of concentrations 25, 50, 75 and 100
ppm were 42.16, 28.4, 20.23 and 15.15 hours, respectively.

3. Toxicity of imidacloprid against different stages of german cockroach B.

germanica
a. Assessing mortalities after 7 days from treatment.
(1) as a stomach poison
The LC50 and the LC90 of imidacloprid against 2nd instar nymphs were
4604 and 16261 ppm, respectively. These values against 6th instar nymphs were
9228 and 24980 ppm, respectively.
The LC50 and LC90 values of imidacloprid as a stomach poison against
adult males were 5441 and 17777 ppm, respectively.
(2) as a contact poison
The LC50 and the LC90 of imidacloprid against 2nd instar nymphs were
3740 ppm and 13108 ppm, respectively. These values against last nymphal
stage were 5646 and 32350 ppm, respectively.
The LC50 and LC90 of imidacloprid as a contact poison against adult
males were 4350 and 14009 ppm, respectively.
b. The mortalities of treated cockroaches after 14 days.
(1) as a stomach poison
For the 2nd instar nymphs, the LC50 and LC90 after 14 days were 1539 and
5413 ppm, respectively. These values against the 6 th instar nymphs were 3656
and 10659 ppm, respectively.

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 The LC50 and LC90 of imidacloprid as a stomach poison against adult
males were 2881 and 10565 ppm, respectively.
(2) as a contact poison
The LC50 and LC90 of imidacloprid against 2nd instar nymphs after 14
days were 920 and 3714 ppm, respectively. These values against the last
nymphal stage after 14 days were 2566 and 11618 ppm, respectively.
The LC50 and LC90 of imidacloprid as a contact poison against adult
males were 1844 and 5090 ppm, respectively.

4. Toxicity of boric acid against different stages of german cockroach B.

germanica.

After 7 days of exposure, the LC50 and LC90 values of boric acid against the
2nd instar nymphs were 8.82 % and 35.38 %, respectively. These values against 6th
instar nymphs were 9.87% and 46.34 %, respectively. The LC50 and LC90 (%) of
boric acid against the adult males were 12.3% and 77.3%, respectively, after seven
days of inspection.

The LC50 and LC90 of boric acid against the 2nd instar nymph After 14 days
of exposure were 3.14% and 11.96%, respectively. These values against the 6th
instar nymph were 4.7% and 17.6 %, respectively. The LC50 and LC90 of boric acid
against adult males were 6.97% and 23.71%, respectively.

The toxicity of boric acid after 14 days could be arranged at a descending

order as follow, 2nd instar nymphs˃ 6th instar nymphs ˃ adult males.
5. The lethal effect (LT50) of boric acid and imidacloprid during 14 days
after treatment.
a. The lethal effect (LT50) of boric acid
The concentrations of 19.0, 25.0 and 37.5 % of boric acid lasted 2.99,
2.63 and 2.48 days to kill 50% of the treated 2nd instar nymphs.

١٦
 The concentrations of 19.0, 25.0 and 37.5 % caused 50 % mortality to 6th
instar nymphs after 3.76, 2.95 and 2.62 days, respectively.
Boric acid lasted 5.42, 3.22 and 2.77 days to kill the half treated adult
males for the concentrations of 19.0, 25.0 and 37.5 %, respectively.
b. The median lethal time (LT50) of imidacloprid
The LT50 of imidacloprid as a contact poison against 2nd instar nymph
after 14 days were 4.72, 2.99, and 1.94 days for the concentrations of 5000,
10000, and 15000 ppm. These values were 5.99, 3.98 and 3.21 days for the
same concentrations of imidacloprid as a contact poison against 6 th instar
nymphs, respectively. The LT50 values of imidacloprid as a contact poison
against adult males were 5.75, 3.25 and 2.65 days, respectively.
The LT50 of imidacloprid as a stomach poison against 2nd instar nymph
after 14 days were 5.30, 3.85 and 3.16 days for the concentrations of 5000,
10000, and 15000 ppm, respectively. These values were 10.65, 5.81 and 3.79
days, respectively, when imidacloprid treated as a stomach poison against 6 th
instar nymphs. These values were 96, 4.28 and 3.38 days, respectively, when
imidacloprid treated as a stomach poison against adult males.
The toxicity of Imidacloprid as a stomach poison could be arranged by
ascending order as follow: 2nd instar nymphs˃ adult males˃ 6th instar nymphs.
The same order of toxicity could be followed when imidacloprid treated as a
contact poison.

6. Joint action of binary mixtures of the three tested insecticides against the
last nymphal stage of German cockroach

The joint action of the three compounds was tested by adding the LC25 of
each according to the following ratio 1:1, 1:2 and 2:1. The co-toxicity factor
was calculated for all mixtures to know their joint action.

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 a. Boric acid and fipronil mixtures:
The mixture of boric acid and fipronil at ratio of 1:1 show a potentiation
effect. On the other hand this mixture showed an additive effect at ratio of 1:2,
but it produced antagonism effect at ratio of 2:1.
b. Boric acid and imidacloprid mixtures:
The ratio of 1: 2 of boric acid+ imidacloprid mixture showed an additive
effect. The other two tested ratios of boric acid and imidacloprid mixtures (1:1
and 2:1) showed an antagonism effect, the co-toxicity factors were -42.86 and -
52.4, respectively.
c. Fipronil and imidacloprid mixtures:
The joint action of the obtained results of that all fipronil and
imidacloprid mixtures at ratios of 1:1, 2:1 and 1:2 led to an antagonism effect.
The co-toxicity factor was calculated for all mixtures to know their joint
action. Out of the nine tested mixtures the fipronil+ boric acid mixture at ratio
1:1 had a potentiation effect. The rest mixtures; two of them had an additive
effect however six mixtures had an antagonism effect.
In conclusion, our findings showed the efficacy of fipronil for control of
cockroaches in laboratory after 24 hrs of exposure. The application of fipronil
bait was not only found more convenient but was also easy to apply (and was
very stable under a wide range of conditions) and there was minimal disruption
of the home or workplaces. Treatment with binary mixture of fipronil with boric
acid 1:1 as a cockroach bait was found to be highly efficacious and safe for the
user and the local environment.

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