INSECTICIDAL EFFICACY OF DIATOMACEOUS EARTH ON LARVAE OF AGRIOTES LINEATUS (COLEOPTERA; ELATERIDAE), POTATO PESTS IN TISSEMSILT, ALGERIA

Abstract

In order to evaluate the diatomaceous earth (DE) toxicity on the Agriotes larva Agriotes line-atus, concentration of 0.1mg/cm²; 0.2; 0.4; 0.6; 0.8 and 1 mg/cm² of DE were directly used in the powder form. Insects were exposed to diatomaceous earth through filter paper placed on Petri dishes at a temperature of 24°±2°C. An amount of 0.2086 of CL50 and 0.2192 mg/cm2 of CL90 were determined. The mortality tests showed that there is a strong correlation between the concentration increase and time increasing. The results showed that the diatomaceous earth can be used as an efficient larvicide against agri-otes, known by their deleterious effects on different cultures.

Full text

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9956
INSECTICIDAL EFFICACY OF DIATOMACEOUS EARTH
ON LARVAE OF AGRIOTES LINEATUS (COLEOPTERA;
ELATERIDAE), POTATO PESTS IN TISSEMSILT, ALGERIA
Bounouira Yassine
1
, Gaouar Benyelles Nassira
1,*
, Senouci Hanane
1
, M'saad Guerfali Meriem
2
1
Ecology and Management of Natural Ecosystems Laboratory, Department of Ecology and Environment, University of Tlemcen, Algeria
2
Laboratory of Biotechnology and Nuclear Technologies, LR16 CNSTN01, National Center of Nuclear Sciences and Technologies,
Technopole Sidi Thabet, Tunis, Tunisia
ABSTRACT
In order to evaluate the diatomaceous earth
(DE) toxicity on the Agriotes larva Agriotes line-
atus, concentration of 0.1mg/cm²; 0.2; 0.4; 0.6; 0.8
and 1 mg/cm² of DE were directly used in the pow-
der form. Insects were exposed to diatomaceous
earth through filter paper placed on Petri dishes at a
temperature of 24°±2°C.
An amount of 0.2086 of CL50 and 0.2192
mg/cm2 of CL90 were determined. The mortality
tests showed that there is a strong correlation be-
tween the concentration increase and time increas-
ing. The results showed that the diatomaceous earth
can be used as an efficient larvicide against agri-
otes, known by their deleterious effects on different
cultures.
KEYWORDS:
Diatomaceous earth, Agriotes, concentration, mortality,
toxicity, larvicide.
INTRODUCTION
Agriotes or wireworms, Agriotes lineatus, are
beetles through which larvae «wireworm» are well
known by their damages. Larvae exclusively live in
the earth where they develop during many years
according to the underground parts of cultivation:
grains, roots, collet and tubers [1].
Due to their underground life mode, wire-
worms can damage the productivity of cereals,
maize, vegetables, small fruits, and ornamental
plants and cause quality deterioration of carrots,
strawberries and potatoes [2, 3], which necessitates
insecticidal treatments in order to control the dam-
ages.
Among the diverse insecticides with retentive
effects, the organophosphorus and the pyrethroids
are the most used [4, 5, 6]. However, the residues of
these synthetic insecticides reduce the acceptability
of the consumer because there is an increasing
demand for foods without residues. Besides the
developing of resistance in the harmful insects to
the insecticides [7, 8], the environmental factors
and health risks necessitate the evaluation of new
substances, rather natural, and less harmful for the
management of the pests. Therefore, the alterna-
tives of chemical insecticides such as diatomaceous
earth are equally evaluated with good efficiency
insecticide [9, 10], the conservatives of the residual
grains continue to play an inevitable role in the
management of the stored pests [11].
The diatomaceous earth (DE) is originally a
natural substance that is certified as an organic
insecticide, which is non-toxic and ecologically
benign [12]. Silica dust of DE is the variable results
which are used against different targeted organisms
[13, 14].
The goal of this study is to evaluate for the
first time the efficiency of the diatomaceous earth
on the mortality of "iron wire". In order to do so,
the powder is sprayed on the larvae with different
concentrations: 0.0 mg; 0.1mg; 0.2mg; 0.4mg;
0.6mg; 0.8mg and 1mg/cm
2
on filter paper, in Petri
dishes of 9 cm in diameter, containing slices of
potatoes in order to guarantee their nourishment.
MATERIALS AND METHODS
Diatomaceous earth (DE) and its treatment
protocol. In our work, we are concerned with a
local material called the diatomite or the diatoma-
ceous earth which is rather abandoned in Algeria.
The diatomite is also called kieselguhr, is a sili-
ceous sedimentary rock, porous and friable made
almost entirely of skeletons of diatomaceous
(Fig. 1).
The diatomaceous or the Bacillariophyceae are
a part of the brown algae junction (Chromophytes).
They are very ancient as the very first fossils go
back to Cretaceous (120 million years) [15]. The
diatomaceous are eukaryotes cells enshrined inside
a hydrated silica wall (the frustula) which are com-
posed of two imbricated units: the epivalve and the
hypovalve. These two valves are linked by connect-
ing belts, composed of fine siliceous bands [16].
The accumulation of a siliceous deposit is
constituted after the degradation of the organic

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9957
matter. The silica deposits of the diatomaceous
fossils constitute a rock called diatomite [17].
The studies have clearly shown that silicon
application was able to contribute in a significant
manner in the reduction of damages caused by
parasites and diseases [18, 19, 20]; In [21].
FIGURE 1
Microscopic structure of the diatomite
Mineralogical analysis by X-ray diffraction
(XRD). The device used in this study is the diffrac-
tometer MiniFlex 600, 40kV, 15mA, scan speed/
duration time 5.0000 deg/min, step width 0.0200
deg, scan range 79.99 deg.
Diatomaceous earth was removed from the
Bider Zone (Tlemcen-Algeria). The composition
of this material was determined by [22] who
showed that it comprises 55.47% SiO2, 6.17%
AL2O3, 13.34% CaO2, 0.87% K2O, 2.3% MgO,
3.29% Fe2O3, PF. at 15.73%, 0.82% Na2O, 0.18%
SO3 and 0.45% TiO2.
Collection and colonization of test insects.
The larvae of agriotes, wireworms (Coleoptera,
Elateridae) were collected at a potato orchard.
Shortly thereafter, the Agriotes were transported in
30 cm x 30 cm x 50 cm plastic containers contain-
ing soil and potato tubers to the laboratory.
For insecticide tests, the insects were exposed
to diatomaceous earth at different concentrations:
0.0 mg; 0.1mg; 0.2mg; 0.4mg; 0.6mg; 0.8mg and
1mg / cm2 on filtering paper in 9cm diameter petri
dishes, containing pieces of potato to feed the lar-
vae.
All the experimental Agriotes were put in the
laboratory at 24 ° ± 2 ° C, and their mortality rate
was monitored every 12h up to 96h.
Statistical analysis. Mortality was calculated
by grouping the number of dead and live beetles at
each replication [23]. Witness mortality was nil and
no correction was necessary.
Repeated measures analysis was performed by
population, with diatomaceous earth treatment as
the main effect, and adult mortality as the response
variable.
The results obtained were subjected to the
analysis of the variance test (ANOVA with 1 classi-
fication criterion). When this analysis reveals sig-
nificant differences, it is supplemented by the Tuk-
eye Kramer (HSD) test [24] at the α = 0.05 thresh-
old.
The lethal concentrations for the diatomaceous
earth were calculated: concentrations that determine
the 50℅ and 90% mortality for the Agriotes popula-
tion. Statistical analysis was performed with SAS
version 9.0.
RESULTS
Mineralogical analysis by X-ray diffraction
(XRD). The results of the mineralogical analysis
done by the XRD are presented in Figure 2.
The XRD rays have shown that the diatoma-
ceous earth which were taken from the Bider zone
(Tlemcen-Algeria) are originally formed of fresh
water containing the HP coesite, silicon dioxide
(SiO
2
) with 77%, Magnesium Oxide (MgO) 4.5 %,
Potassium Oxide (K
2
O) 1.0 %, Calcium Oxide
(CaO) 6.8 %, sulfur (VI) oxide (SO
3
) 0.6 %, phos-
phorus (V) oxide (P
2
O
5
) 1.0 %, iron(III) oxide,
hematite HP (Fe
2
O
3
) 4.1 %, Vanadium Phosphide
(PV) 0.4 %, Phosphorus Sulfide (P
4
S
7
) 1.0 %, Tita-
nium Oxide (TiO
2
) 0.2 %, Aluminum Oxide
(Al
2
O
3
) 2 %, Phosphorus (P) 1,0 % and Sodium
Oxide (Na
2
O) 0.4 %.
The XRD analysis has also shown that our di-
atomite sample contained 4 phases: the SiO
2
Amor-
phe with 32.19%, Ankerite in the form of Ca (Fe
+2
,
Mg) (CO
3
)2 with 31.9%, Calcite CaCO
3
with
26.9% and Quartz SiO
2
with 9.0%.

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9958
2-theta (deg)
Intensity (cps)
20 40 60
0
1000
2000
FIGURE 2
X-rays of diffraction obtained on our diatomite samples.
TABLE 1
Physic-chemical analyses of diatomaceous earth sample.
pH Electrical conductivity
µm/cm 1/5
organic
matter %
Limestone % assimilable
phosphorus ppm
total Active
7,61
12360
0,17
18,75
4,75
245,78
TABLE 2
ANOVA test for the mortality of larvae of ladybugs treated with diatomaceous earth (DE)
at 0.1mg / cm2; 0.2; 0.4; 0.6; 0.8 and 1 mg / cm 2.
Source DDL Anova SS F Pr > F
time
6
109.2108844
16.25
< 0.0001
Dose
6
451.1156463
67.11
< 0.0001
Repetition
2
0.0544218
0.02
0.9760
Statistical analysis was performed with SAS version 9.0.
TABLE 3
Susceptibility of populations of Agriotes larvae at different concentrations of diatomaceous earth (DE)
Dose Tukey Groupement Average mortality in each replicate
1
A
8,46
0,8
A
8,08
0,6
B
5,7
0,4
B
4
0,2
C
0,18
0,1
C
0
0
C
0
The physic-chemical analyses of our sample
have shown that the level of pH was 7.61, its con-
ductivity was 12360 µm/cm 1/5 and the percentage
of organic matter was 0.17. This diatomaceous
earth contains 18.75% and 7.75% total limestone
and active, respectively and 245.78 ppm of assimi-
lable phosphorus (Table 1).
Efficient DE concentrations. The main ef-
fects on the levels of mortality of the tested popula-
tion were significant. The tested diatomite was
toxic to the individuals of agriotes population.
According to the results of the analysis, the
mortality variance of agriotes population was high-
ly significant according to doses and time (F=67.11,
P< 0.0001, F=16.25, P< 0.0001, respectively) (Ta-
ble 2).
We know from previous work that different
strains of the same species have different suscepti-
bilities to DE [25], that the concentration of DE
affects mortality and that the DEs differ in their
efficiencies [26].

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FIGURE 3
Cumulative mortality rate of Agriotes larval populations exposed to diatomaceous earth (DE) at 0.1mg /
cm
2
; 0.2mg / cm
2
; 0.4mg / cm
2
; 0.6mg / cm
2
; 0.8mg / cm
2
and 1 mg / cm 2 during 96 hours. The values are
the averages of three repetitions.
TABLE 4
Estimated values of lethal concentration
(CL) after diatomaceous earth (DE) treatment of larval beetles
Mortality percentage (%) lethal dose (mg/cm
2
)
01
0.1880
10
0.1985
20
0.2021
30
0.2046
40
0.2067
50
0.2086
60
0.2105
70
0.2126
80
0.2152
90
0
.2192
99
0.2314
To compare the efficacy of different concen-
trations of diatomaceous earth, a Tukey test was
performed (Table 3), a similar efficacy was found
between the 1mg and 0.8mg / cm2 concentrations
with a mortality rate of 100 % in less than 60 hours
of exposure; between 0.4 and 0.6 mg/ cm2 with a
mortality rate of 100% in less than 96 hours of
exposure and between 0.1 and 0.2 mg/ cm2 a near-
zero mortality rate after 96 hours of exposure. The
control at 0 mg/ cm2 had a zero mortality rate after
96 hours of exposure.
Our study gives the opportunity to directly
compare the sensitivity of the Agriotes population
to our DE formulation; comparisons were based on
observed mortality. It would be interesting to know
the lethal concentrations to control this population.
Estimation of lethal concentration (LC)
values for the agriotes population. We calculated
the lethal concentrations to determine diatomaceous
earth concentrations that determine mortality for the
Agriotes population over time. The table 4 shows
the lethal concentrations for our diatomaceous
earth, it allowed us to conclude that the diatoma-
ceous earth (DE) has a very important toxic effect
on the population of Agriotes at the 0.2086 LC50
and the CL90 of 0.2192 mg / cm2.
Cumulative mortality. The mortality curve of
the Agriotes during the 96h of exposure remains nil
for the controls and the concentration 0.1 mg / cm2;
for the concentration 0.2 mg / cm 2 the mortality
reaches 20% in 96 hours of exposure. As concentra-
tions increase, mortality reaches 100% with de-
creased exposure times; 96h for the concentration
0.4 mg / cm2, 84h for the concentration 0.6 mg /
cm2, 60h in the concentration 0.8 mg / cm2 and 48h
for the concentration 1 mg / cm2 (Fig. 3).
DISCUSSION
The effectiveness of DE in controlling stored
product pests depends on various factors such as
insect species, products, moisture, and temperature
[26].
[25] found that insects of the same species but
of different origins had different levels of
susceptibility to DE.
The effectiveness of the natural insecticide DE
varies according to the geological origin of the
mines from which it is extracted [27], probably

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because of the differences in the physical and
morphological properties of diatoms [28].
The effects of DE on a wide range of beetles
have been reported by many studies, including
Rhyzopertha dominica [26, 10], Tribolium
castaneum [25, 29], Tribolium confusum [30],
several Sitophilus species [31, 26, 30],
Callosobruchus maculatus [31], and Plodia
interpunctella [30].
Other than from the beetles mentioned above,
DE was used as a feed additive to control internal
parasites and fly larvae, including house flies,
sedentary flies and flies in manure [32].
In addition, it has been reported to be effective
against cockroaches, silverfish, bed bugs, mites,
ants and fleas [33], gastropods such as slugs [34]
and also against infestations [35].
To understand the basis of variability, it was
necessary to determine the dust's mode of action.
Different theories have been proposed: (1)
superficial enlargement of the integument following
dehydration [36]; (2) alteration of the digestive tract
[37]; (3) blockage of spiracles and tracheas [38]; (4)
ad- or lipid absorption of the cuticle [39] and / or
(5) damage to the protective wax layer [40]. It is a
mechanical insecticide, insects cannot develop
immunity or resistance to DE. Therefore, it can be
used to control insects for a long time without the
manifestation of insecticide resistance that is often
reported for other insecticides
CONCLUSION
These results allow us to suggest this natural
product as a plausible alternative to insecticides
previously used to fight against Agriotes lineatus,
and this will be better to preserve our health and
environment.
ACKNOWLEDGEMENTS
The authors thank Gérard DUVALLET
professor emeritus at the University of Paul-Valéry
of Montpellier for his help, and we thank
HAOUALA Rabiaa Professor at the Higher
Agronomic Institute of Chott Meriem of Tunisia for
helping us with the corrections.
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Received: 15.08.2019
Accepted: 18.09.2019
CORRESPONDING AUTHOR
Gaouar Benyelles Nassira
Ecology and Management of
Natural Ecosystems Laboratory,
Department of Ecology and Environment,
University of Tlemcen – Algeria
e-mail: gaouarn@yahoo.fr