The control of greater wax moth,

Galleria mellonella L., by chemicals

1. Susceptibility of the wax moth larvae and honey-bee workers to
certain chemicals’

By A. D. ALP, N. M. BAKRY’, M. A. ABDELLATIF4

s.
and K. EL-SAWAF’

With I Figure

Abstract

Toxicity of the 13 insecticides (allethrin, cidial, dimethoate, dioxathion, endosulfan, endq-

sulfan-fenitrothion, ethion, Fac, imidan, imidan-endosulfan, pyrethrins, toxaphene, tri-
chlorfon) mixed with the artificial diet on the third larval instar of Gallaeria mellonella
are determined.
Toxicities of the same insecticides on larvae fed on comb pieces dipped in these insecti-
cides are obtained.
The effects of the same insecticides when applied topically on newly emerged honey-bee
workers 24 hours after treatment are evaluated.
The residual effect of the insecticides in treated comb pieces after storage for different
periods are determined. The 120-hour mortality percentages of third instar larvae of combs
treated with LC;, concentration a n d stored for 6 months are: 25 for trichlorfon, 35 for
endosulfan, 43 for dimethoate and dioxathion, 50 for endosulfan-fenitrothion and Imidan-
endosulfan, 63 for toxaphene, 65 for allethrin and cidial, 68 for Fac, 70 for ethion, 80 for
pyrethrins and 90 for Imidan. These results indicate that most insecticides used in treating
the combs are quite toxic to the larvae and having a long residaual action.

Introduction

There are several species of moths which damage the wax combs either in
storage or in hives. The greater wax moth, Gulleviu mellonellu L., is the most
serious of them all, since its larvae may cause considerable damage. This
pest is well adapted to live in hives and weak colonies in warm regions.
Tropical and subtropical areas of low latitudes suffer greatly from its in-
festation. If colonies become weak through failing queens, application of
insecticides, or other reasons the combs are likely to be damaged or destroyed

Contribution of thc Plant Protccrion Dept., Faculty of Agriculture, Alexandria Uni-

versity, Egypt.
Plant Protection Dept., Ministry of Agriculture, Baghdad, Iraq.
:’ Assistant Prof. of Pesticide Chemistry, Fnc. of Agric.. Ales. Univ., Egypt.
’ Associate Prof. of Entomology, Fac. of Agric., Ales. Univ., Egypt.
Prof. of Entomology, Fac. of Agric., Alex. Univ., Egypt.
 T h e control of greater w a x moth, Galleria mellonella L., by chemicals 171

within a relatively short time. Combs stored in a building, unless properly

fumigated, are apt to be destroyed by the greater wax moth larvae.
This moth lays its eggs in crevices in the wooden body of the hive out of
reach of the bees. The newly hatched larvae burrow into the comb, forming
tunnels. The older larvae eat out the combs converting them completely
into frass, and the silken web they spin sticks neighbouring combs together
(Fig.).
Fumigation (KREBS1957) is the only practical method still used by
beekeepers in the apiaries to control different stages of this pest, especially
during early infestation to eliminate the loss in combs. Many fumigants are
used in this respect, such as carbon disulphide, sulfur, calcium cyanide, me-
thyl bromide, ethyl dibromide, ethylene oxide, and par2 dichlnrobenzene.

A comb heavily infested with the greater w a x moth

However, these fumigants will not prevent infestation, besides some are
harmful to man and bees. Moreover, they need to be applied several times
to the combs in storage to ensure their protection against any possible in-
festation.
There has been always a need for a new method of protection which
would be harmless to either bees or man, as well as being very effective
against the greater wax moth. The present work deals with evaluating some
chemical against this pest as well as on honey-bee workers, in hope to
develop a better, easier, safer effective method of control.

Materials and methods

1. Chemical insecticides

The following insecticides were used throughout the course of this work:
Technical 90 O/u and a formulated 40 O / o emulsifiable concentrate of dimethoate-0,0-
Diniethyl S-(N-niethylcarbamoylinethyl) phosphorodithioate;
172 A. D . A l i , N.M . Bakry, M . A. Abdellatif and S. K . El-Sawaf

Technical 95 Oil0 and formulated 20 O / o emulsifiable concentrate Fac(R)-0,O-Diethyl S-(N-

isopropylcarbamoylmethyl) phosphorodithioate;
Technical 90 O / o and formulated 50 O / o emulsifiable concentrate of cidial(R)-Ethyl mer-
captophenylacetate, 0,O-dimethyl phosphorodithioate;
Technical 90 O / o and formulated 35 O / o emulsifiable concentrate of endosulfan-6,7,8,9,10,
l0-Hexachloro-1,5,5a,6,9,9a-hexachloro-6,9-methano 2,4,3 benzodioxathiepin 3-oxide;
Technical 95 O / o of ethion-O,O,O’,O’-tetraethyl S,S’-methylenephosphorodithioate;
Technical 90 O / o of pyrethrins;
Technical 90 O / o of allethrin;
Technical 90 O / o and formulated 47 O / o emulsifiable concentrate of dioxathion-S,S’-p-
dioxane-2,3-diyl 0,O-diethyl phosphorodithioate (cis and trans isomers) ;
Technical 97.3 o/o of Imidan-0,O-dimethyl-S-phthalimidomethylphosphorodithioate;
Technical 90 Q / o and formulated 60 O/o emulsifiable concentrate of toxaphene-chlorinated
ramphene containing 67-69 O / o chlorine;
90 O / o wettable powder of trichlorfon-0,O-dimethyl (2,2,2-trichloro-l-hydroxyethyl)
phosphonate;
Endosulfan-fenitrothion mixture: emulsifiable concentrate containing 300 gm./L. endo-
sulfan, a n d 600 mg/l fenitrothion. Fenitrothion is -0,O-dimethyl 0-(4-nitro-m-tolyl) phos-
phorothioate;
Imidan-endosulfan mixture, wettable powder containing 30 “/o endosulfan and 20 O / o
Imidan.

2. Bioassay tests

Feeding w a x moth larvae on treated artificial diet

Solutions of insecticides were made by dissolving a certain weight of technical insecticide
in aceton. Different concentrations for each insecticide were made t o assess for susceptibility
levels (LC,, and LC,,). O n e ml of each concentration was added t o 5 grams of the artificial
diet (HAYDAK 1936); which consists of 4 parts fine corn meal, 2 parts whole wheat flour,
2 parts skim-milk powder, 2 parts standard wheat middling, and 1 part dried yeast, and
equal parts of this d r y diet and a liquid mixture containing one part honey and one p a r t
glycerine were mixed), and mixed thoroughly. These treatments were carried out in Petri-
dishes incubated a t 30’ C. Mortality counts were recorded 24 hours after treatment.

Feeding w a x m o t h larvae on treated w a x c o m b pieces

Pieces of combs of about 4 >.< 6 cm each were prepared. Technical insecticides were dissolved
in acetone (except for endosulfanfenitrothion and Imidan-endosulfan where formulated
mixtures were used). 200 ml of each concentration was put in a 403 ml beaker and the comb
pieces were dipped for one minute. The treated comb pieces were then left to d r y for 24
hours. Larvae of Galleria ntellonella were put on each piece in a Petri-dish and incubated
a t 30” C. Mortality counts were recorded 24 hours afler treatment.

Topical application of insecticides on h o n e y bees

i l d u l t honey-bee workers (1-2 day old) were treated individually by applying one micro-
liter drop of acetone solution of the technical insecticide on the dorsal surface of the thorax
(GRAVESand MACKENSEN 1965). The insecticide-acetone solutions were applied with a
micro syringe. Treated bees were placed in small cages supplied with candy and water and
kept at 3 0 ’ C. Mortality counts were made 24 hours after treatment.

Detevmination of residual toxicity of dipped c o m b pieces using

w a x m o t h larvae
A test was carried monthly for 6 months period on treated comb pieces t o determine the
decrease in toxicity by storage. Acetone solutions of technical insecticides were made at
the LC,,, and LC!,,, levels. Small comb pieces were dipped in 200 ml of every concentration
for one minute, dried and stored i n conditions similar to that in storage. The residual toxi-
city was determined by placing a treated comb piece in a Petri-dish with third instar
iarvae, and mortality counts were taken after 24, 72 and 120 hours after treatment.
 T h e control of greater w a x moth, Galleria rnellonella L., by chemicals 173

Results and discussion

Toxicity of insecticides mixed with artificial diet t o Galleria larvae

It is obvious from Table 1 that Imidan-endosulfan and endosulfan-feni-
trothion were the most toxic. Their LCBOvalues amounted to 0.0105 and
0.018 and LCgo values to 0.040 and 0.076 mg/g diet, respectively. However,
endosulfan alone was slightly less toxic than both mixtures. The slightly
higher toxicity of them could be due to a potentiation effect resulting from
the presence of an organophosphorous insecticide (Imidan in the first and
fenitrothion in the second).
Table 1
LC50 and LCgo values of the insecticides used against the third instar larvae of
Galleria rnellonella fed treated artificial diet

I Insecticide
LC50
mg per
g diet
LC80
mg per
g diet

Alethrin 0.420 1.500

Cidial 0.058 0.190
Dimethoate 0.210 0.390
Dioxathion 1.400 3.000
Endosul fan 0.034 0.090
Endosulfan-fenitrothion 0.018 0.076
Ethion 0.700 5.400
Fac 1.600 3.300
Imidan 1.600 3.000
Imidan-endowl fan 0.011 0.040
Pyrethrins 0.092 0.260
Toxaphene 0.680 3.600
Trichlor ion 0.240 0.500
Control - -

Cidial follows endosulfan having LCx and LCw of 0.058 and 0.19 mg/g
diet, respectively. Pyrethrins came after. Dimethoate and trichlorfon showed
almost the same level of toxicity.
Alethrin was far less toxic than pyrethrins, with LCjo and LCw of 0.42
and 1.50 gm/g diet, respectively. Both toxaphene and ethion gave nearly the
same level of toxicity.
The other three insecticides, dioxathion, Imidan and Fac were the least
toxic among compounds tested.
The above results do not show clear relationship between the toxicity of
any compound and the group of insecticides to which the compound belongs.

Toxicity of dipped comb pieces in different insecticides t o

Galleria larvae
The same insecticides were used to evaluate their insecticidal protecting
action on wax combs. Dipping pieces of comb in insecticides solutions were
made at various levels of concentrations and lor one minute.
The results in Table 2 show that endosulion-fenitrothion was the most
toxic, with L Cx of 0.001 and LCw of 0.005 "/n.
174 A. D. A l i , N . M . Bakry, M . A . Abdellatif and S.K . El-Sawaf

As it is clear from Tables 1 and 2, the insecticides showed almost the

same descending pattern of toxicity in both methods, although the con-
centrations according to the LC values were different for some insecticides.
Endosulfan and Imidanendosulfan showed almost the same levels of tox-
icity.
Table 2
L C d s and LCgo’s of the insecticides used against the third instar larvae of
Galleria mellonella fed treated old comb pieces for one minute

Insecticide LCjo (percent) LCBo(percent)

Alethrin 0.580 1.050

Cidial 0.008 0.034
Dimethoate 0.250 0.400
Dioxathion 0.700 1.900
Endosul fan 0.002 0.010
Endosulfan-fenitrothion 0.001 0.005
Ethion 1.880 4.300
Fac 1.030 1.520
Imidan 0.010 0.060
Imidan-endosulfan 0.003 0.012
Pyrethrins 0.270 0.960
Toxaphene 0.004 0.012
Trichlorfon 0.080 0.150
Control - -

The differences found in the toxicity pattern between the two methods
are expected. I n the dipping method the amount of insecticide absorbed by
the wax comb pieces will depend on the lipophilic property of each in-
secticide. This means, the more the lipophilicity, the more the amount ab-
sorbed of insecticide.

Toxicity of insecticides to honey-bee workers

Susceptibility of the honey-bee workers to the insecticides were determined.
Table 3 shows that cidial and dimethoate are quite toxic with LDx’s of
0.05 and 0.07 and LDso’s of 0.09 and 0.10 ug./bee, respectively. Following
them Imidan and pyrethrins, their LDJ,’s are 0.12 and 0.25, and their
LD IU’S are 0.17 and 0.90 ug./bee, respectively.
Imidan-endosulfan and endosulfan-fenitrothion are having quite similar
toxic action and toxicity is almost twice that of endosulfan alone. However,
the toxicity of Imidan-endosulian could be due to Imidan alone knowing
that the mixture at the LDio level contains the same amount of the LDw of
Imidan alone (0.12 ug./bee). Also, the amount of endosulfan present at
the LDsl level of the mixture does not add any toxicity. The former six
insecticides could be classed in one group with a LDx range from 0.05 to
0.3 u g h e e and considered relatively toxic group.
Endosulfan alone follows the former group with LDx and LDw of 0.65
and 1.10 ug./bee, respectively. Fac and ethion show similar toxic effect with
LD-)o’sof 0.8 and 0.85 and LDw’s of 1.25 and 1.60 ug.’bee, respectively.
Trichlorfon and allethrin are less toxic having LDiu’s of 1.2 and 1.56 and
LDtIll’sof 1.6 and 2.6 ug./bee, respectively. The five former insecticides with
 The control of greater wax moth, Galleria mellonella L., by chemicals 175
Table 3
LDso’s and LDgo’s of the insecticides applied topically against newly
emerged honey-bee workers

Insecticide LD,, ug./bee LDg, ug./bee

Alethrin 1.560 2.600

Cidial 0.050 0.090
Dimethoate 0.070 0.100
Dioxathion 4.300 7.300
Endosulf an 0.650 1.100
Endosulfan-fenitrothion 0.306 0.648
Ethion 0.850 1.600
Fac 0.800 1.250
Imidan 0.120 0.170
Imidan-endosulfan 0.300 0.550
Pyrethrins 0.250 0.900
Toxaphene 3.700 5.800
Trichlorfon 1.200 1.600

LDjo ranging from 0.65 to 1.56 ug./bee could be considered moderately toxic
group.
Toxaphene and dioxathion are the least toxic of the group. Their LDao
values are 3.7 and 4.3 and LDgo values are 5.8 and 7.3 ug./bee, respectively.
Both insecticides are considered relatively non-toxic.
Laboratory screening tests by ANDERSON and ATKINS(1958 and 1967)
and TODDand MCGREGER (1960) showed the insecticides could be tabu-
lated according to their toxicity to honey bees into 3 groups. The first group
included highly toxic pesticides, and such compounds are not used in this
work. The second group included moderately toxic pesticides such as ethion
and endosulfan. The third group included relatively nontoxic pesticides such
as allethrin, pyrethrins, toxaphene, trichlorfon and dioxathion. Most of
the insecticides used in this work were chosen from the third nontoxic group.

Toxicity of the insecticides residue in treated stored

c o m b pieces t o Galleriu larvae
ARer evaluating the effect of the insecticides on the larvae it was found
necessary to determine the residual effect of these insecticides in treated
combs.
The data in Table 4 represent the residua1 toxicity given as percent
mortality 24,72 and 120 hours after treatment, for treated comb pieces at the
LCno and LCgo levels (Table 2), stored at different periods O and 6 months.
The results show a decrease in toxicity by storage for 24-hour percent
mortality at both concentration levels, LCX and LCw. However, the de-
crease is not pronounced for longer time of administration especially for the
LCWconcentration after 6 months of storage.
The 120-hour mortality percentages for combs treated with the LCX con-
centration and stored for 6 months are 25 for trichlorfon, 35 for endosulfan,
43 for dimethoate and dioxathion, 50 for endosulfan-fenitrothion and Imi-
dan-endosulfan, 63 for toxaphene, 65 for allethrin and cidial, 68 for Fac,
70 for ethion, 80 for pyrethrins and 90 for Imidan. In other words the
insecticides which give 50 or less percent mortality 12O-hours after treatment
176 A. D. Ali, N.M . Bakry, M . A . Abdellatif and S. K . El-Sawaf
Table 4
Residual toxicity data for treated comb pieces at LC5o and LCgo levels against
third instar larvae of GuUeriu meUoneUu L.

LCso level LCgo level

Storage-
Insecticide period 010 mortality for different @Inmortality for different
hours after treatment hours after treatment
24 72 120 24 72 120

Alethrin 0 46 83 100 95 100 -

6 8 20 25 30 45 65
Cidial 0 55 95 100 90 100 -
6 33 58 80 58 80 100
Dimethoate 0 52 85 100 95 100 -
6 35 70 90 78 95 -
Dioxathion 0 57 80 100 90 100 -
6 20 45 65 58 78 100
Endosulfan 0 42 68 90 85 100 -
6 5 20 35 25 45 68
endowl fan-fenitrothion 0 50 85 100 95 100 -
6 23 38 50 30 45 70
Ethion 0 45 78 100 88 100 -
6 23 45 75 45 70 90
Fac 0 55 78 95 88 100 -
6 20 43 68 48 73 100
Imidan 0 52 85 100 95 100 -
6 35 70 90 78 95 100
Imidan-endosulfan 0 47 65 85 85 100 -
6 16 35 50 40 65 78
Pyrethrins 0 55 95 100 90 100 -
6 33 58 80 68 80 100
Toxaphene 0 50 85 100 95 100 -
6 33 45 63 68 90 100
Trichlorfon 0 45 83 100 95 100 -
6 8 20 25 30 45 65

for 6 months storage are trichlorfon, dimethoate, dioxathion, endosulfan,

endosulfan-fenitrothion, and Imidan-endosulfan. And those which give more
than 50 O / o are pyrethrins, Imidan, cidial, Fac, ethion, allethrin and toxaphene.
These results indicate that most insecticides used are quite toxic to the
larvae and have a long residual action. Although these insecticides could give
a very good protection to combs in storage, they could not be recommended
for usage inside the hive after storage time. Thus recommendation of such
insecticides might be limited to those with shorter residual action.

Zusammenfassung

Zur Bekampfung dcr Grollen Wachsmotte Galleria mellonella L. mit Chemikalien

I . Empfindlichkeit d e r Wachmottenraupen sowic Honigbienen auf bestimmte Chemikalien
Es wurde die Toxizitat von 13 Insektiziden (Allethrin, Cidial, Dimethoate, Dioxation,
Endosulfan, Endosulfan-fenitrothion, Ethion, Fac, Imidan, Imidan-endosulfan, Pyrcthrine,
Toxaphen, Trichlorfon) gemischt mit der kiinstlichen Diat bei der L:4-Raupe von G . m e l k -
nella bestinimt. Fur dieselben Insektizide wurde dic Toxizitat fur die an naturlicher N a h -
rung frosenden Raupen fcstgestellt. Des weiteren wurde die Wirkung dieser Imektizide
auch auf Honigbienen-Arbeiterinnen 24 Std. nach der Behandlung dcr Waben ermittelt.
 The control of greater wax moth, Galleria mellonella L., by chemicals 177

Schliealich wurden die Ruckstandewirkungen der Insektizide in den behandelten Waben fur
bestimmte Zeiten untersucht. 6 Monate nach Behandlung der Waben zeigten die Bienen fol-
gende Mortalitatswerte: 25 O/o bei Trichlorfon, 35 O / o bei Endosulfan, 43 O / o bei Dimethoate
und Dioxathion, 50 O/o bei Endosulfan-fenitrothion und Imidan-endosulfan, 63 O / o bei Toxa-
phen, 65 O / o bei Allethrin und Cidial, 68 O / o bei Fac, 70 O / o bei Ethion, 80 O / o bei Pyrethrine
und 90 O / o bei Imidan. Diese Ergebnisse zeigen, da8 die meisten Insektizide sowohl fur die
Schadlingsraupen als auch fur die Bienen sehr toxisch sind und einen langen Riickstands-
effekt zeigen.

References

ANDERSON, L. D.; ATKINS,E. L. jr., 1958: Toxicity of pesticides to honeybees in laboratory

and field tests in Southern California. J. Econ Entomol. 51, 103-108.
ATKINS,E. L. jr.; ANDERSON, L. D., 1967: Toxicity of pesticides to honeybees in the
laboratory. Intern. Apicult. Congr., Zlst, Ann. Rev. Entomol. 13, 213-231.
GRAVES,T. B.; MACKENSEN, O., 1965 : Topical application and insecticide resistance studies
on the honeybees. J. Econ. Entomol. 58, 990-993.
HAYDAK, M. H., 1936: A food for rearing laboratory insects. Jour. Econ. Ent. 29, 1026.
KREBS,H. M., 1957: Ethylene-dibromide-death to the wax moth control. Clearing in Bee
Culture 90, 682-684.
TODD,F. E.; MCGRAGER, S. E., 1960: The use of honeybees in the production of crops.
Ann. Rev. Entomol. 5, 265-278.

Aus dem Institut fur Angewandte Zoologle der Universitat Wurzburg

Untersuchungen uber den Einflufi des Beutetiers auf den

Erbeutungsvorgang bei der Waldameise, Formica polyctena
Foerster (Hymen., Formicidae)
Von K. BUTTNER

Mit 17 Abbildungen

Abstract

Studies on the influence of the prey on predation by the wood ant, Forrnira polytena rcerst.
The influence of factors such as mobility, size, consistence, defensive behaviour and
pattern of the prey on the predation of insects by Fovmica polyctena is investigated. The
phase of orientation is studied by means of the time and direction which the ants need for
orientation; the phase of fight by means of the intensity of the attacks, the number of
attacking ants, the duratioq of attacking actions and by the degree of spurted formic acid;
the phase of transport by the speed and manner of transportation. In addition to cxperi-
ments with living insects studies with sham models of the same size as the prey were
carried out.

1. Einleitung

Die Beziehungen eines Raubers zu seiner Beute werden meist einseitig von
der Seite des Raubers untersucht. Insbesondere bei der Kleinen Waldameise

%. a n g . E n t . 74 (19731, 177-196
&> 1973 Verlng P a u l Parey H a m b u r g u n d Berlin
ASTXI-Cuden: ZANFAF.