Proceedtngs of the 7th International Worktng Conference on Stored-product Protection - Volume 1
The relative toxicity of phosphine to eggs of the Angonmois
grain moth Sitotroga cerealella (OLIV .) (Lepidoptera:
Gelechiidae) and the almond moth Ephestia cautella
(Walker) (Lepidoptera: Pyralidae)
Mohamed E1 Hassan Shaza1i1 and Christoph Relchmuth2
Abstract
Investigations were carried out at 25"C and 70% r. h. to
assess the toxicity of phosphine to eggs of Sitotroga
cerealella (OLIV.) and Ephestia cautella (Walker). Three
phosphine concentrations (0. 1 mg/L, 0.5 mglL and 1. 0
mglL) , four egg age groups (one, two, three and four days
old) with 16 h, 24 hand 48 h exposure periods,
respectively, were tested.
The eggs of S. cerealella were found to be more tolerant
than the eggs of E. cautella over the range of exposure
periods and concentrations tested. Increase of exposure
period was more effective than mcrease of gas
concentrations to achieve complete mortality. The ct-
products for 95 % mortality of the two to four days old eggs
of S. cerealella ranged between 4 mg PH3 hlL and 40 mg
PH3 hlL and were higher than the values for the
corresponding eggs of E. cautella (3 mg PH3 hlL to 14 mg
PR3 hlL). One day old eggs required higher concentranons
and times for complete control.
Introduction
Sttotroga cerealella, commonly known as the Angoumois
gram moth, ISa very destructive insect pest of cereal grams
in the trOPICSwhere its development is faster. Eggs are
usually laid singly among the grains or may be laid together
in groups fastened together The eggs are rmlky white, pear
shaped, pomted anterioraly and rounded posterioraly. On
hatching, the larvae enter a grain of It choice and feed and
develop internally.
On the other hand, Ephestia cautella, commonly known
1 Gram Storage Research Unit, Shambat Research Station,
Box 30, Khartoum North, Sudan
2 Federal Biological Research Centre for Agnculture and Forestry,
Institute for Stored Product Protection, Korugm-Luise-Strape
-14195 Berlm, Germany
as the almond moth, IS a serious pest of many food
commodities m temperate regions. The eggs are creamy in
appearance and sphencal in shape They are laid singly and
on hatching the larvae feed externally.
Hydrogen phosphide is a commonly used fumigant for the
control of stored product insects However, the toxicity of
phosphine to insects vanes greatly with temperature
(Lindgren et aI., 1958); the species (Hole et aI., 1976)
and the developmental stage (Bell, 1976). The differences
between the tolerance of stages are extremely great (Howe,
1973) .
The immature stages, particularly the eggs and pupae are
reported to be more tolerant than adults. However, newly
laid eggs are most tolerant against phosphine for most of the
stored product insects.
The wide variation in tolerance of phosphme to the eggs
of some moths was reported by Bell (1976). The tolerance
of eggs of Rhyzopertha dominica (FAB.) and
Prostephanus truncates Horn was Investigated by Hashem
and Reichmuth (1989).
However, there have been no studies on the efficiency of
phosphine on eggs of S. cerealella. The present study was
designed to assess and compare the toxicity of phosphine to
the eggs of S. cerealella compared to E. cautella.
Materials and methods
Insect culture
The insect stocks used for the investigations are kept in
culture since many years at the Institute for Stored Product
Protection, Federal BIological Research Centre for
Agriculture and Forestry, Berlin, Germany.
To obtain the eggs, the adult moths of S. cerealella and
E cautella were reared on whole wheat kernels and wheat
bran, respectively, In 2 liter glass Jars. The Jars were
tightly covered with muslin cloth, kept in place by rubber
P 0
bands, and were kept m a CTH-room at 22"C and 10% r. h.
The laboratory stocks were cultured weekly.
Egg collection
19, D
About 100 adults of each species were collected by sucking
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 Proceedings of the 7th Internatwnal Working Conference on Stored-product Protection. - Volurne 1

through a rubber tube into a SIeve placed inside a desiccator occurred. The cnterion for egg mortality was failure to
for egg laying. Air current was produced by a small pump. hatch into a larvae. Unhatched eggs, of both species, dry
After 24 h the eggs were collected in the bottom of the out and wnnkle. Treatments were replicated three times.
SIeve. With the atd of a bmocular microscope, 50 eggs Untreated controls were set up to each treatment.
batches were counted and placed on sticky paper stnps, 5 By use of the computer program TABLECURVE® and
ern long and 1 em Wide. WIre mesh cages 5.5 em m length unlmear regression L~s values were determined and in
and 1.5 cm in diameter, were used to contam the egg some cases (beyond 48 hours of lethal exposure, and less
stnps, which were kept in the expenmental room at 22"C than O. 1 mg PH31L or more than 1 mg PH31L lethal
and 10% r. h. until required for fumigation. concentration) extrapolated
Production of phosphine
Results
Phosphine was generated from magnesium phosphide
powder reacting to 100 % phosphine. The powder reacts The results of mortality of eggs of different ages, for both
WIth water according to the formula: moths, with different phosphine concentrations over 16 h,
M~P2 + 6H20 = 3Mg( OHh + 2PH3 25 hand 48 h exposure penods at 25"C and 10% r. h. are
The pnnciples of the standard FAO procedure (Anonym, presented m the Table 1.
1975) was followed The method provided a convenient At the highest tested concentration (1 mg/L) and 48 h
source of phosphine for dosing purposes over a penod of exposure nearly complete mortality of eggs occurred.
time, depending on the rate of removal. However, some one day old eggs of S. cerealella survived.
Fumigation chamber The eggs of E. cautella required, for 100 % mortality, 48
hat 1 mglL for one day old eggs, 48 hat 0.5 mglL for two
The fumigation vessels used consisted of Dressel flasks days old eggs, 48 hr at 0.1 mglL for three and four days old
(0.7 1) with ground glass stoppers. A recirculatory 3 flask eggs compared with 48 h at 1 mglL for two days old eggs
apparatus was set up to provide small fumigation chambers and 48 h at 0.5 mglL for three and four days old eggs. At
for one concentration and three exposure periods. The flasks 48 h exposure the mortality beyond 80% for one and two
were connected together and to a gas collecting tube of 1 .2 I days old eggs also occurred at a higher concentration for S.
by PVC tubing A small electric pump was set up to cerealella than E. cautella. The mortalities resulting from
recirculate the gas evenly throughout the apparatus. A each of the exposure penods of 16 h, 24 hand 48 h
silicon rubber septum fitted m the narrow tube, protruding increased With age in both species.
from the gas reservoir, was used to mject and Withdraw It is obvious (FIgure 1 and Figure 2) that the
phosphine with gas tight Hamilton syringe. Just before corresponding ct-products (LC9s x Ltgs) for eggs of a certain
injection, a volume of arr equal to that of phosphine to be age of S. cerealella are higher at all exposure periods than
injected was Withdrawn from the reservoir. The calculated for the eggs of E. cautella. ThIS implies that the eggs of
volume of gas, for each dose, was withdrawn from the gas S. cerealella are more tolerant to phosphme than the eggs
burette and than injected mto the gas reservoir, through the of E. cautella. Also Table 2 and Table 3 show this
self sealing rubber septum.
tendency. The Ltgs for 0.1 mg PH31L and one day old eggs
The time of injection was noted and the pump operated to of S. cerealella and for 1 mg PH31L and three and four
distribute the gas. After 10 minutes the concentration of days old eggs of E. cautella could not be extrapolated due
phosphine was determined twice by a phosphme gas to lack of sufficient data in the range (see Table 1, Table 2
analyser.
and Table 3 as well as FIgure 1 and Figure 2) Obviously,
The glass flasks containing the eggs were then the tolerance of two days old eggs of S. cerealella drops
disconnected without loosing any gas and Immediately below the values for one day old eggs of E. cautella for
transferred to the CT-room at 25"C , where they were kept concentrations greater than 0.6 mg PH31L, indicating the
through the exposure penod,
strong sensitivity towards phosphine in this age group. The
Post fumigation procedure LD9S values for three and four days old eggs of E. cautella
fall together at lower concentrations (Figure 2). At all
At the end of the exposure period the flasks were taken to
exposure penods, the ct-products to obtain 95% mortality
the fume cupboard and their lids removed for aeration. The
increased as the period of exposure decreased from 48 h to
small cages, con taming the egg strips, were taken out and
16 h (Table 4). It can be inferred that longer exposures
placed m Petri dishes and transferred on trays in the CT-
cause higher mortality than short ones, if the ct-product is
room at 25"C and 70% r. h. Observations on egg mortality
kept constant.
started a week later and continued until no further hatches

327
 Proceedings of the 7 th International Working Conference on Stored-product Protection - Volume 1

100

~~t
I ! : I I i
90 l..jfo
<,

-.
.- Silolroga,'~ day old ,
e::s 80 '" I
-1\1--- Ephestla, 1 day old .1

0
70 -,
i
-+- Sitotroqa, 2 days old'+-f-
.c !
I

--*- Ephestla, 2 days old L

.5
~ 60
1_____
" ~-~r~~f t , f-

'~---n
+4+I-H
0 4~~ __ I
.t: ""
i

50 ---.
I
Q)
Q. Jr\ i ~

'~:"
I
~
e~ 40 J -.;~
--II I
rn
0
Q.
30 I ~
~~
"',-, ~
<.
-~
>< ~ I",~ ,'1111
I
---+-+-
w 20

10
I
I
i

I
'" "X
\
~t-+
i
f--+-

I i
~ .--

II
o
1 ! i I i I

0.01 0.10 1 00 10.00

Concentration in mg PH~I

Fig. 1. LD:.15 for one and two days old eggs of Sitotroga cerealella and Ephestza cautella and furmgation WIth phosphme at 2S'C
(The German program used 'mstead of' m the Ioganthrmc scale of the graph)

100
! I

90 I I

--.- Sitotroqa, 3 days old

I
80 ___ Ephesna, 3 days old

e:::J 70 -+-- Sitotroqa, 4 days old

0 -)E- Ephestia, 4 days old
..c :
.: 60
I
'tl
0
'C
~\ I
I
Q) 50 ---
Q. ,

e:::J 40
I
~~ ~ :
If)
<; ~ I
8-
><
w 30 n <, ~ -. I ! I

20
I

,
!

I
,
~-~-
'S~
<;

;----..
~
--..
~'-... '~
'r--1 ,"'.
\,

~..
~t
I I
i
-~-I---

i
10
~--..ll,'" I i
--
I

o I I I I i I
001 010 100 10 00
Concentration in rng PH:II

Fig. 2. L~5 for three and four days old eggs of Sitotroga cerealella and Ephestza cautella and fumigation WIth phosphme at
2S'C (The German program used 'mstead of' m the loganthrmc scale of the graph).

328
 Proceedings of the 7th International Worktng Conference on Stored-product Protectum - Volume 1

Table 1. Mortality of eggs of different age of Sitotrox: Table 3. LUJsin mg PH3IL to control one day, two days,
cerealella and Ephestia cautella after fumigation three days and four days old eggs of Sttotroga
at 25°C With phosphine with different cerealella and Ephestia cautella with fumigation
concentrations and exposure periods. at 25°C.

Mortality m % LUJs
Exposure in
Concentration of PH3 m mglL Exposure period m hours Egg age in days
hours
0.1 0.5 1.0
1 2 3 4
S. cerealella
1 day old eggs 16 3.3 6.1 12.8 Sitotroga cerealella
1 day old eggs 24 7.4 20.5 32.9 16 >10 2.45 1.18 0.88
1 day old eggs 48 16.5 85.0 95.9
24 5 -10 0.99 0.89 0.48
2 days old eggs 16 28.2 47.2 62.7
2 days old eggs 24 46.9 78.7 95.1 48 0.99 0.45 0.12 0.09
2 days old eggs 48 78.6 95.7 100 Ephestia cautella
3 days old eggs 16 51.0 64.1 85.6
16 >2.3 0.85 0.48 0.29
3 days old eggs 24 76.1 85.2 97.9
3 days old eggs 48 91.8 100 100 24 2 .3 0 .41 0 . 13 0 . 13
4 days old eggs 16 64.1 81.4 98.5 48 0.4 0 .08 < 0 . 1 < 0.1
4 days old eggs 24 90.0 95.1 100
4 days old eggs 48 95.9 100 100 Table 4. ct-products in mg PH3 hIL to control (LD9S) one
E. cautella
day, two days, three days and four days old eggs of
1 day old eggs 16 9.6 43.3 67.9 Sitotroga cerealella and Ephestui cautella with
1 day old eggs 24 16.5 77.2 84.2 fumigation at 25"C for 16 hours, 24 hours, or 48
1 day old eggs 48 80.0 96.5 100 hours, respectively.
2 days old eggs 16 71.3 87.1 97.9
2 days old eggs 24 86.1 95.7 100 Ct-product in mg PH3 hIL
2 days old eggs 48 97.1 100 100 Egg age in days
3 days old eggs 16 81.8 95.3 100
3 days old eggs 24 90.7 100 100 1 2 3 4
3 days old eggs 48 100 100 100 Sitotroga cerealella
4 days old eggs 16 84.0 97.9 100
16 39.2 18.9 14.1
4 days old eggs 24 93.1 99.3 100
4 days old eggs 48 100 100 100 24 23.8 21.4 11.5
48 21.6 5.8 4.3

Table 2. L1i)sof eggs of different age of Sitotroga cerealella Ephestia cautella

and Ephestw cautella for phosphme fumigation at 16 13.6 7.7 4.6
25"C With 0.1 mg/L, 0.5 mgIL and 1 mg/L, 24 55 .2 9.8 3. 1 3.2
respectively.
48 19.2 3.8
L1i)sm days
Concentration of PH3 in mgIL Discussion
0.1 0.5 1.0 The results confirmed the known tendency of high tolerance
S. cerealella of young eggs of stored product pest insects to phosphine
1 day old eggs > 90 53.3 47.5 (FIgure 1 and FIgure 2). However, it is for the first time
2 days old eggs 88.7 43.8 23.9 that this has been shown for one, two, three and four days
3 days old eggs 54.3 33.6 20.1 old eggs of S. cerealella in comparison to E cauiella .
4 days old eggs 45 23.8 11. 7 Bell and Glanville (1970) demonstrated that the eggs of
E. cautella Ephestia eluiella (HuBNER), E. kuehmieiui (ZELLER),
1 day old eggs 52.5 39.6 35.5 E. cautella ( WALKER) and Plodw interpunctella
2 days old eggs 38.5 22.8 11.4 ( HuBNER) were tolerant to phosphme at 25"C, If the
3 days old eggs 31.2 15.7 < 16 exposure did not exceed two days. Similar tolerance to 24 h
4 days old eggs 31.1 11.2 < 16 exposure to phosphine has been found by other researchers
329
 Proceedings of the 7th Internatwnal Working Conference on Stored-product Protection - Volume 1

for E. cautella (Baskaran and Mookherjee , 1991; Muthu, Anonym 1975. Recommended methods for the detection and
1973). Bell (1976) indicated the position and length of measurement of resistance. 14. Tentative method for
penod during which eggs of E. cautella, E. kuenniella ; adults of some major pest species of cereals, With methyl
E. elutella and P. interpunctella are tolerant to bromide and phosphme. FAO Plant Protection Bulletin.
phosphine at a range of temperatures. However, the 23, 12 - 25.
mortality obtained after short exposures was not always Baskaran, P. , and Mookherjee , P. B. 1971. Effect of food
proportional to concentration, probably different levels of on the susceptibility of Cadra cautella WALKER and
susceptibihtyoccur during the tolerant phase and variations Trogoderma granarium EVERTS to phosphine. Indian
in the exact age of eggs in different age groups may be Journal of Entomology 33, 23 - 39.
critical. Bell, C. H. 1976. The tolerance of developmental stages of
Bell (1976) suggested that the high tolerance to four stored product moths to phosphme. Journal of stored
phosphine m the egg stage is linked with metabohsm durmg Products Research. 12, 77 - 86.
the development of the embryo. The consumption of oxygen Bell, C. H., and Glanville, V 1973. The effect of
during the egg stages varies in different species concentration and exposure m tests with methyl bromide
(WIgglesworth, 1972). Bond (1968) showed that oxygen IS and phosphine on diapausing larvae of Ephestia elutella
essential for phosphine to be effective and stages which can (HuBNER) (Lepidoptera: Pyrahdae ). Journal of stored
survive without oxygen for a period of time are likely to be Products Research. 9, 165 - 170.
tolerant However, available data are not sufficient to Hashem, M. Y. , and Reichmuth, Ch. 1981 The efficiency
correlate low oxygen consumption and high tolerance to of phosphine against eggs of lesser gram borer
phosphine during the egg stage. The high efficiency of Rhyzopertha dominica (FAB.) and larger grain borer
phosphine after longer than after shorter exposures was Prostephanus truncatus (HORN) (Coleoptera:
attributed to continued insect development while under Bostrycludae ). Nachrichtenblatt des Deutschen
fumigation and that more time mcreases the chances of Pflanzenschutzdienstes. 41, 159 -163.
reaching a susceptible stage (Bell, 1976). Hole, B. D. , Bell, C. H. , Mills, K A., and Goodship, G.
1976. The toxicity of phosphme to all developmental stages
Acknowledgements of thirteen species of stored product beetles. Journal of
stored Products Research. 12, 235 - 244.
The DAAD (German Academic Exchange Service) is thanked Howe, R. W. 1973. The susceptibility of the immature and
for financial support of Dr. Shazali's short visit to the adult stage stages of Sitophilus granarius to phosphine.
Institute for Stored Product Protection in Berlin, Germany. Journal of stored Products Research. 8, 241 - 262
The authors are very grateful to MISS Verena Lemke for Lindgren, D. L., Vincent, L. E., and Strong, R. G.
technical assistance in takmg and analysis of gas samples. 1958. Studies on hydrogen phosphide as a fumigant
We also wish to thank Mrs. Sabine Berger for collectmg the Journal of Economic Entomology 51, 900 - 903.
eggs of different test insects. Muthu, M. 1973. Some aspects of phosphine as a fumigant.
In: S. K Majumder and J. S. Venugopal (Eds.):
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330