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Laboratory Rearing of Immature Stages of Oryctes

agamemnon arabicus under Three Constant Temperatures

Article · June 2012

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Regional centre for Agriculture research, Sidi Bouzid
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 Laboratory Rearing of Immature Stages of Oryctes
agamemnon arabicus under Three Constant Temperatures

Rasmi Soltani, Centre Régional de Recherches Agricoles de Sidi Bouzid, 9100 Sidi
Bouzid, Université de Kairouan, Tunisie

__________________________________________________________________________
ABSTRACT
Soltani, R. 2012. Laboratory rearing of immature stages of Oryctes agamemnon arabicus under
three constant temperatures. Tunisian Journal of Plant Protection 7: 35-42.

Oryctes agamemnon arabicus was a new exotic species signalled in the oasis of southwest Tunisia
since 1995 where it was specifically associated to date palm. Few bibliographies on the biology of the
species exist in the world. Thus, the aim of this work is to study the development of immature stages of
this pest under laboratory conditions at three temperatures 23, 27 and 30°C. Results concerning the
development of immature stages showed that means duration of the incubation period, the total larval
development and pupa were 14.16 ± 1.69, 227.62 ± 15.79, and 24.29 ± 2.75 days at 23°C, 11.84 ± 1.3,
199.2 ± 12.32, and 22.27 ± 1.92 days at 27°C, and 9.49 ± 1.17, 222.39 ± 10.43, and 21.16 ± 1.85 days
at 30°C, respectively. These facts indicated that O. agamemnon arabicus accomplished its total
immature stage development in 266.18 ± 15.96 days at 23°C, 229.93 ± 10.35 days at 27°C, and 253.03
± 10.94 days at 30°C.

Keywords: Date palm, life cycle, Oryctes agamemnon arabicus, rearing, temperature.
__________________________________________________________________________

Date palm, Phoenix dactylifera, is The rhinoceros beetle Oryctes

a strategic crop of economic importance
in Tunisia. It is widely grown
successfully in the south of the country;
its cultivated area occupies about 40,000
ha with around 6 million trees (2).
Locally, wide range of insect species are
able to attack date palm trees causing a
significant and serious yield losses; the
most harmful are the carob moth
Ectomyelois ceratoniae, the white scale
Parlatoria blanchardi, Apate monachus,
the acarina called Boufaroua Oligonychus
afrasiaticus and others (2).
Corresponding Author: Rasmi Soltani
Email: rasmi.soltani@iresa.agrinet.tn
Accepted for publication 14 March 2012
Tunisian Journal of Plant Protection
agamemnon arabicus (Coleoptera,
Scarabaeidae) is signaled as a secondary
pest within orchards of date palm trees in
most Gulf countries as Kingdom of Saudi
Arabia (18), Irak, United Arab Emirates
(12), Sultanate of Oman (1) and lately in
Tunisia (9). This pest had been officially
signaled in Tunisia for the first time in
1995 after the sudden collapse of many
productive palm trees in Mrah Lahouar
oasis from Tozeur governorate (9, 13,
14). Inside oasis, it was considered as a
monophagous pest attacking only date
palm tree with its different varieties. After
this date, the pest was reported in several
other locations of the Djerid and on the
oases of Rjim Maatoug belonging to
Kebili governorate. Until 2006, the pest
invaded about 3,310 ha of date palm trees
located in Djerid zone and in the extreme
west of Kebili (15).
35 Vol. 7, No. 1, 2012 
 Development of immature stages
occurred inside hidden and well protected
breeding sites of the palm tree making its
management difficult. These breeding
sites are located almost on different part
of the palm tree mainly on respiratory
roots and external components of the
stem (matrixes of fibrilium, stem bark and
inside the basal part of dry petiole) and
the four oldest levels of green palms
situated at the base of the crown. After
several years of repeated attacks,
specifically on respiratory roots part, this
pest can causes significant damage that
result in the collapse of the plant as
already observed in Mrah Lahouar and
Rjim Maatoug (15, 16).
Little data are available on O.
agamemnon arabicus both in Tunisia and
worldwide. In order to develop better
techniques for its monitoring and
management strategies, understanding the
life history of this pest can constitute a
first basically step and a mainly key to
develop such techniques. Thus, the
present work aimed to study the duration
of developmental period of immature
stages of this pest under three controlled
temperatures conditions.
MATERIALS AND METHODS
Biological material. Adults of O.
agamemnon arabicus used in this study
were originally collected from infested
area of Rjim Maatoug in Kebili
governorate, located on the southwest of
Tunisia during the end of summer 2006.
A number of sixteen couples were used in
this survey. Females used in this study
started already their oviposition activity
in the field. After their catch, they were
also confined with male inside
rectangular plastic boxes (15 × 20 × 10
cm). These boxes were filled with a thick
layer of 3-5 cm of friable material,
collected from natural breeding sites
within oases, in the goal to create a
Tunisian Journal of Plant Protection
middle like inside infested parts of palm
trees where couples burrowed to copulate.
It is also important to mention that couple
formation, copulation, and oviposition
activity occurred inside obscure parts of
the plant.
Breeding and experimental
protocol. Breeding medium used in all
assays of this work was originally chosen
by Soltani et al. (17) to breed the same
species under laboratory conditions. This
substrate is of natural origin, collected
directly from the living sites of the
species in date palm trees. It consists of
fine wood granules, which are the bits of
food discarded on both sides of the
mouthparts of larvae during their diets on
the wood of hairy respiratory roots or on
dry petiole and fibrilum matrixes, of less
than 3 mm in size.
Initial relative humidity measured
for initially collected substrate depends
on the existing developmental stage. In
fact, from oviposition sites, it varies from
26 to 34% while for the larvae sites this
moisture ranges between 38 to 65%.
This substrate was mainly used for
females which burrowed themselves
inside to lay eggs on. The first part of the
study which interested couples was
carried under ambient temperature of the
laboratory. Boxes were daily controlled
twice (in the morning and in the
afternoon); during these operations
content of each box was sieved separately
to collect fresh eggs laid inside the
substrate. Then, substrate was returned
inside box with female to continue its
oviposition activity.
Eggs collected in the same time,
never directly handled, were transferred
together, by group of 10 maximum, to
small rectangular opaque plastic boxes (5
× 10 × 5 cm) used as incubator and
containing a 3 cm thick layer of the same
friable material used for oviposition. Eggs
36 Vol. 7, No. 1, 2012 
were two third burrowed on the surface of
substrate to facilitate developmental
survey; then boxes were moved under
controlled conditions. Experiences were
initially established using 70, 100 and 60
eggs in the same previous order of
temperature. Holes were made in the
cover of each box to decrease the
humidity inside.
Larval development occurred
through three stages which differentiation
occurred based on the dimensions of
cephalic capsules and the body size as
already determined by Soltani et al. (17).
They were reared by group of three larvae
inside opaque plastic boxes (15 × 20 × 10
cm) on the same food substrate used
previously. Boxes were filled with a thick
layer of substrate of 5-7 cm. Larvae
reared inside the same box are of the
same age i.e., they hatched or molt in the
same day.
Rearing of O. agamemnon
arabicus occurred in the laboratory within
three different obscure climate chambers
and at three constant temperatures: 23 ±
1°C, 27 ± 1°C and 30 ± 1°C. Humidity
depended on the breeding stage; in fact,
incubation took place under a mean of
relative humidity of 30 ± 4%. However,
all larval development and pupae
occurred under an average of relative
humidity of 50 ± 8%. Photoperiod, which
mainly interest larvae, was nil because
they are blind without eyes.
Biological parameters. Pest
development was daily examined until
adult emergence. The following
parameters were surveyed:
- The incubation time: time
elapsed between egg laying and egg
hatching.
- The larval development: larval
development completed through three
distinct stages including pre-pupa
(including the third stage). The duration
Tunisian Journal of Plant Protection
of each one of them was measured.
During experiments, specimens that died
were discarded but the duration of each
biological stage completed before death
was used in the calculation of means.
Each one of the three larval stages was
studied separately, then the total duration
of the larval development was estimated.
- Pupa: time elapsed between third
larval molting and adult emergence.
Once development of these
instars achieved, the developmental
period of immature instars was
determined for each temperature.
Statistical analyses. Collected
data were subject to statistical analyses
using the SPSS software for windows
(version 16). Analyses of variance
(ANOVA) applied to each stage
permitted testing the effect of
temperatures on the period of incubation,
larval development and pupae. Means
were compared and separated into subsets
using the LSD test at 5% level.
RESULTS
Immature stage development.
The mean duration of development of
immature stages (measured in days) at
various constant temperatures with
standard deviation are given in Table 1.
Incubation. The incubation
period, as illustrated in Table 1,
constituted the shortest stage of
development of the pest. In fact, eggs
were hatched in 14.16 ± 1.69, 11.84 ± 1.3
and 9.49 ± 1.17 days at 23, 27 and 30°C,
respectively. Analysis of variance showed
significant differences between
incubation periods under the tested
temperatures. The LSD-test allowed the
classification of means into three
homogeneous subsets represented by the
letter (a, b, and c) inside Table 1.
37 Vol. 7, No. 1, 2012 
 Table 1. Developmental time of immature stages of Oryctes agamemnon
arabicus reared in laboratory at three constant temperatures regimes
Days required (mean ± SD) to complete each life stage
23°C 27°C 30°C
Initial eggs Ni = 70 Ni = 100 Ni = 60
14.16 ± 1.69 a 11.84 ± 1.3 b 9.49 ± 1.17 c
Incubation
n = 61 n = 87 N = 53
35.42 ± 3.23 a 34.85 ± 3.13 a 35.78 ± 2.27 a
L1
n = 48 N = 78 N = 45
Larval 50.82 ± 4.87 a 43.66 ± 3.88 b 45.19 ± 3.14 c
L2
stages N = 38 N = 74 N = 42
140.85 ± 15.23 a 118.22 ± 10.05 b 141.84 ± 11.78 a
L3
N = 34 N = 69 N = 39
24.29 ± 2.75a 22.27 ± 1.92 b 21.16 ± 1.85 c
Pupa
N = 34 N = 55 n = 38
Means in the same line followed by the same letter (a, b or c) are not
significantly different according to the LSD test (at P ≤ 0.05,); Ni: Initial
number.

The infertile eggs are recognizable
four days after oviposition by their color
turning on brownish and their aspect
becoming dry and shriveled. However,
alive eggs changed their volume
progressively and their color initially
white tends to transparence when
hatching approached. The registered
hatching rates were 87.14% at 23°C, 87%
at 27°C and 88.33% at 30°C. These
results showed a high rate of fertility
under these different temperatures.
Larval development. The mean
duration of the first and the second larval
stages were illustrated in Table 1.
Statistical analysis using the one-way
ANOVA reveal the presence of a
significant differences for the second
larval stage only; the relative means were
classified in three distinct subsets based
on the LSD-test (Table 1).
The third larval stage completed
its development in 140.85 ± 15.23, 199.2
± 12.32 and 141.84 ± 11.78 days at 23°C,
27°C and 30°C, respectively. Analysis of
variance showed significant differences
between these periods (Table 1). The use
of LSD-test permitted the classification of
temperature means into two subsets as
follows: 23°C/30°C and 27°C.
Tunisian Journal of Plant Protection
The larval period relative to each
temperature varied between minimum
and maximum of 193-274, 173-228 and
201-245 days at 23, 27 and 30°C,
respectively, with respective means of
227.62 ± 15.79, 199.2 ± 12.32 and 222.39
± 10.43 days (Figure 1). Statistical
analysis revealed the existence of a
significant difference between these
periods of development under the
different temperatures; means were
separated into two subsets i.e. 23°C/30°C
and 27°C.
Advanced interpretation of these
results revealed that the third larval stage
dominates both the larval development
and the total development of immature
stages. In fact, it represented 61.88%,
59.35% and 63.78% of larval
development and 52.92%, 51.42% and
56.06% of immature stages duration at
23°C, 27°C and 30°C, respectively.
Before achieving the pre-pupae
stage, larvae migrate deeply inside the
substrate and reduce progressively their
feeding activity. Their migration was
stopped by the basal and lateral inner
sides of the box, which is hard and
difficult to perforate, stimulating, thus,
the entry to pre-pupal stage. In fact, each
larva kept its position inside the room
38 Vol. 7, No. 1, 2012 
situated at the extremity of its feeding
tunnel, repels the substrate using its legs
and compresses it using its abdominal
extremity; by this way, walls of the room
became more cemented and compact. The
volume of the room was estimated to four
times the volume of the larva. At the end,
larva gains a dorsal position; the legs
suspended in the air and initiate the pre-
pupal period.
Pre-pupa. Pre-pupa constitute an
intermediate pseudo-stage between the
larval life and pupa. In fact, larvae which
gain a dorsal position become sluggish
wrinkled with suspended feeding and
movement (legs in air). The duration of
this pseudo-stage was not noticed in
Table 1; it was merged with the duration
of third stage. However, it lasted on
average 17.91 ± 2.45 days at 23°C, 15.86
300
a
250
b
Duration (days)
200
150
100
50
0
Larval Development
Fig. 1. Duration of the larval development and the complete cycle of immature stages of Oryctes agamemnon
arabicus recorded under three constant temperatures (Means of the same group followed by the same letter (a, b
or c) are not significantly different according to the LSD test (at P ≤ 0.05)).
Tunisian Journal of Plant Protection
± 2.86 days at 27°C and 15.84 ± 2.72
days at 30°C.
Pupa. Pupa took minimum and
maximum period of 19-29, 18-27 and 17-
25 days at 23°C, 27°C and 30°C,
respectively. The use of ANOVA-test
reveals a significant difference between
the registered periods which were divided
into three subsets based on the LSD test
(Table 1).
The total mean durations of
immature stages relative to each
temperature were illustrated in Figure 1
with larval development. It shows a total
domination of larval development as
compared to the other stages (Table 1);
the relative percentage of this duration
ranged between 85.51% and 87.89%
under these temperatures. Analysis of
variance showed a significant difference
between these periods (Figure 1).
a
c
a b
23°C
27°C
30°C
Complete Cycle
39 Vol. 7, No. 1, 2012 
 As shown in Figure 1, the
immature stage developmental duration
varies depending on temperatures tested.
Significant differences were observed in
total life cycle of immature stages of O.
agamemnon arabicus developed under
the three constant temperatures. In fact,
the duration was longer when the insect
was reared at 23°C and 30°C in
comparison with 27°C. Means durations
were 266.18 ± 15.96, 229.93 ± 10.35, and
253.03 ± 10.94 days at 23°C, 27°C and
30°C, respectively. This result was
confirmed by the LSD-test which allowed
the classification of temperature means in
three homogeneous subsets (Figure 1).
Table 1 also shows that incubation period,
the second and third larval stages and
pupa contributed significantly to this
difference.
DISCUSSION
As shown in above presented
results, the immature stages
developmental time varies greatly
depending on temperatures. In fact, the
optimum temperature for breeding,
permitting the lowest duration of
development, is situated at 27°C. It was
clear that the immature stages and
particularly the larval development
dominates the life cycle of this pest.
Generally, this cycle is similar to that of
other species belonging to the same genus
characterized, as shown in Table 2, by the
dominance of larval developmental time
as compared to the duration of the total
cycle.
In the present study, it is important
to mention that the total life cycle of this
pest was completed in less than 8 months
at 27°C and less than 9 months both at
23°C and 30°C. Thus, O. agamemnon
arabicus could produce three
generations/two years. In comparison to
the bibliography, it is mentioned that the
developmental cycle within Oryctes
genus was well diversified (Table 2).
However, some species have less than
one year life cycle as reported for O.
rhinoceros (11, 18) while others have one
year life cycle as is the case of O. elegans
(11) and O. agamemnon (10). The
development of O. nasicornis occurs
either on one year in Ivory Coast or on
two to three years in Ukraine (3). Life
cycle duration in this case was dependant
on existing conditions.
Obtained results at 23°C were
similar to those reported by Soltani et al.
(17) under 23°C and a RH of 55 ± 6 %
which advanced a mean developmental
time of 274 days for O. agamemnon on
the same substrate.

Table 2. Developmental time of three species of the genus Oryctes

Time required in days
Larval development Pupa References
Species Eggs
L1 L2 L3
10-
115-175 20-28 (10)
O. 16
agamemnon 11- 31- 51-
98-162 20-29 (13, 17)
16 38 68
11 21 21 35-63 21 (8)
O. 10- 12-
8-12 60-165 17-28 (4, 5, 19)
rhinoceros 21 21
12 72-130 20 (6)
O. elegans 2-8 270-300 21 (11)

Tunisian Journal of Plant Protection 40 Vol. 7, No. 1, 2012 

 Concerning O. agamemnon constitute a basic step in further
arabicus collected from Tunisia, larvae understanding the species. In fact, it
represented the harmful stage of the pest confirmed in one hand the mastery of
within date palm oases (13, 14, 16). breeding techniques and on the other
Consequently, the knowledge of such hand provided important and fundamental
biological information will constitute in information on its developmental and
the future a base to start a management immature stages cycle under three
program against this pest. constant temperatures. By this way, it
In conclusion, the present work became possible to start management
focused on the immature stages of O. experiments on this pest under laboratory
agamemnon arabicus obtained under conditions before being applied in the
controlled laboratory conditions oases ecosystem.
__________________________________________________________________________
RESUME
Soltani R. 2012. Elevage au laboratoire du scarabé rhinocéros Oryctes agamemnon arabicus sous
les conditions de trois températures constantes. Tunisian Journal of Plant Protection 7: 35-42.

Oryctes agamemnon arabicus est une espèce exotique qui a été signalée pour la première fois dans les
oasis du sud ouest de la Tunisie en 1995 et où elle se trouve inféodée au palmier dattier. Peu de
bibliographies sur la biologie de l’espèce existent à travers le monde. D’où, l’objectif de ce travail est
d’étudier le développement des stades immatures de l’espèce sous les conditions du laboratoire et à
trois températures constantes 23, 27 et 30°C. Les résultats concernant le développement des stades
immatures montrent que les durées moyennes obtenues pour la période d’incubation, le développement
larvaire complet et la nymphose sont de 14.16 ± 1.69, 227.62 ± 15.79 et 24.29 ± 2.75 jours à 23°C, de
11.84 ± 1.3, 199.2 ± 12.32 et 22.27 ± 1.92 jours à 27°C et de 9.49 ± 1.17, 222.39 ± 10.43 et 21.16 ±
1.85 jours à 30°C. Ces résultats indiquent qu’O. agamemnon arabicus accomplit son cycle de
développement jusqu’à l’émergence du stade imago en une durée moyenne de 266.18 ± 15.96 jours à
23°C, 229.93 ± 10.35 jours à 27°C et 253.03 ± 10.94 jours à 30°C.

Mots clés: Cycle de vie, élevage, Oryctes agamemnon arabicus, palmier dattier, température
__________________________________________________________________________
‫ﻣﻠﺨﺺ‬
‫ ﺗﺤﺖ ﺛﻼث‬،Oryctes agamemnon arabicus ‫ اﻟﺘﺮﺑﻴﺔ اﻟﻤﺨﺒﺮﻳﺔ ﻟﺨﻨﻔﺴﺎء وﺣﻴﺪة اﻟﻘﺮن‬.2012 .‫ رﺳﻤﻲ‬،‫ﺳﻠﻄﺎﻧﻲ‬
Tunisian Journal of Plant Protection 7: 35-42. .‫درﺟﺎت ﺣﺮارة ﺛﺎﺑﺘﺔ‬

‫ﻼ أﺷﻴﺮ إﻟﻰ وﺟﻮدﻩ ﻓﻲ واﺣﺎت اﻟﺠﻨﻮب‬ ً ‫ ﻧﻮﻋًﺎ دﺧﻴ‬Oryctes agamemnon arabicus ‫آﺎﻧﺖ ﺧﻨﻔﺴﺎء وﺣﻴﺪة اﻟﻘﺮن‬
‫ وﺗﻌﺪ اﻟﻤﺮاﺟﻊ اﻟﻌﻠﻤﻴﺔ اﻟﻤﻬﺘﻤﺔ ﺑﺸﺄن‬.‫ ﺣﻴﺚ ارﺗﺒﻂ وﺟﻮدهﺎ ﺑﺸﻜﻞ ﺧﺎص ﺑﻨﺨﻴﻞ اﻟﺘﻤﺮ‬1995 ‫اﻟﻐﺮﺑﻲ اﻟﺘﻮﻧﺴﻲ ﻣﻨﺬ ﺳﻨﺔ‬
‫ ﻓﺈن اﻟﻬﺪف ﻣﻦ هﺬا اﻟﻌﻤﻞ هﻮ دراﺳﺔ ﺗﻄﻮر اﻟﻤﺮاﺣﻞ ﻏﻴﺮ اﻟﺒﺎﻟﻐﺔ ﻟﻬﺬﻩ اﻵﻓﺔ‬،‫ ﻟﺬا‬.‫ﺑﻴﻮﻟﻮﺟﻴﺔ هﺬا اﻟﻨﻮع ﻧﺎدرة اﻟﻮﺟﻮد ﻓﻲ اﻟﻌﺎﻟﻢ‬
‫ ﺑﻴﻨﺖ اﻟﻨﺘﺎﺋﺞ أن ﻣﻌﺪل ﻓﺘﺮة اﻟﺤﻀﺎﻧﺔ‬.‫س‬°30 ‫ و‬27 ‫ و‬23 :‫ﺗﺤﺖ اﻟﻈﺮوف اﻟﻤﺨﺒﺮﻳﺔ ﻋﻨﺪ ﺛﻼث درﺟﺎت ﻣﻦ اﻟﺤﺮارة‬
‫ ﻳﻮﻣﺎ‬2.75 ± 24.29 ‫ و‬15.79 ± 227.62 ‫ و‬1.69 ± 14.16 ‫واﻟﺘﻄﻮر اﻟﺸﺎﻣﻞ ﻟﻠﻴﺮﻗﺎت واﻟﻌﺬارى آﺎﻧﺖ ﻋﻠﻰ اﻟﺘﻮاﻟﻲ‬
± 222.39 ‫ و‬1.17 ± 9.49 ‫س و‬°27 ‫ ﻳﻮﻣﺎ ﻋﻨﺪ‬1.92 ± 22.27 ‫ و‬12.32 ± 199.2 ‫ و‬3.1 ± 11.84 ‫س و‬°23 ‫ﻓﻲ‬
‫ وﺗﺸﻴﺮ اﻟﻨﺘﺎﺋﺞ أﻳﻀﺎ إﻟﻰ أن ﻓﺘﺮة ﺗﻄﻮر اﻷﻃﻮار ﻏﻴﺮ اﻟﺒﺎﻟﻐﺔ ﻟﻠﺨﻨﻔﺴﺎء ﺗﺪوم‬.‫س‬°30 ‫ ﻳﻮﻣﺎ ﻋﻨﺪ‬1.85 ± 21.16 ‫ و‬10.43
‫ ﻳﻮﻣﺎ ﻋﻨﺪ‬10.94 ± 253.03 ‫س و‬°27 ‫ ﻳﻮﻣﺎ ﻋﻨﺪ‬10.35 ± 229.93 ‫س و‬°23 ‫ ﻳﻮﻣﺎ ﻋﻨﺪ‬15.96 ± 266.18 ‫ﻓﻲ اﻟﻤﻌﺪل‬
.‫س‬°30

Oryctes agamemnon arabicus ،‫ ﻧﺨﻴﻞ اﻟﺘﻤﺮ‬،‫ دورة ﺣﻴﺎة‬، ‫ درﺟﺔ ﺣﺮارة ﺛﺎﺑﺘﺔ‬،‫ ﺗﺮﺑﻴﺔ ﻣﺨﺒﺮﻳﺔ‬:‫آﻠﻤﺎت ﻣﻔﺘﺎﺣﻴﺔ‬
__________________________________________________________________________

Tunisian Journal of Plant Protection 41 Vol. 7, No. 1, 2012 

LITERATURE CITED
1. Al-Sayed, A.E. and Al-Tamiemi, S.S. 1999.
Seasonal activity of the fruit stalk-borer,
Oryctes agamemnon (Burm.) (Coleoptera-
Scarabaeidae) in Sultanate of Oman. Egypt. J.
Agri. Res. 77: 1597-1605.
2. Anonyme, 2010. C.R.D.A Tozeur, 2010. Rapport
d’activités.
3. Balachowsky, A.S. 1962. Entomologie appliquée
à l’agriculture. Tome I. Coléoptères. Premier
Volume. Caraboidea- Staphylinoidea-
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