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A meridic diet for laboratory rearing of red palm weevil, Rhynchophorus

ferrugineus (Coleoptera: Curculionidae)

Article in Scientific Research and Essays · October 2013

DOI: 10.5897/SRE2013.5502

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 Vol. 8(39), pp. 1924-1932, 18 October, 2013
DOI: 10.5897/SRE2013.5502
Scientific Research and Essays
ISSN 1992-2248 © 2013 Academic Journals
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Full Length Research Paper

A meridic diet for laboratory rearing of Red Palm

Weevil, Rhynchophorus ferrugineus (Coleoptera:
Curculionidae)
H.A.F. El-Shafie1*, J.R. Faleiro2, M.M. Abo-El-Saad1 and S.M. Aleid1
1
Date Palm Research Center of Excellence, King Faisal University, Al-Ahsa 31982, P.O. Box 400, Saudi Arabia.
2
Mariella, Arlem –Raia, Salcette, Goa-403 720, India.
Accepted 28 May, 2013

The red palm weevil (RPW) Rhynchophorus ferruginous (Olivier) was fully developed and reared on a
meridic diet consisting of agar, distilled water, commercial yeast as well as laboratory produced amino
and fatty acid rich brewer’s yeast (Saccharomyces cerevisiae), wheat meal, corn flour, benzoic acid,
ascorbic acid, sorbic acid, vitamin mix and tetracycline hydrochloride. A group of weevils reared on
date palm trunk under the same laboratory conditions was used as control. Diet reared female weevils
laid fertile eggs which successfully hatched into healthy larvae in 3.6 days. Between the first and last
(eighth) larval instars there was a 2311 fold increase in the larval body biomass before pupation. Linear
regression analysis involving log of head capsule width of each instar and instar number resulted in a
2
straight line (R = 0.978) indicating that Dyar's rule is applicable in the case of RPW. Larval development
was completed in 43 to 47 days while the pupa reached the adult stage in 31 to 38 days with an average
of 35 day. The whole life cycle of the weevil from egg to adult, was completed in 78 to 85 days. The
average adult longevity for male and female weevils was 83.3 and 74.8 days, respectively. Besides the
above, important biological parameters including pre-oviposition period, incubation period, per cent
egg hatchability, fecundity and larval weight gain were recorded. A food-fiber pupation technique was
developed with 100% pupation efficiency.

Key words: Rhynchophorus ferrugineus, meridic diet, biological parameters, food-fiber pupation technique,
larval instar, Dyar's rule.

INTRODUCTION

The red palm weevil (RPW), Rhynchophorus ferrugineus planting material shipped for farming and landscape
(Coleoptera: Curculionidae), is an internal tissue borer gardening. In the Middle-East RPW was first reported
reported to have infested 26 palm species worldwide in from Rass-El-Khaima in the UAE in 1985 (Zaid et al.,
diverse agro-ecosystems (Malumphy and Moran 2009 2002) and reached the Eastern region of the Kingdom of
http://www.defra.gov.uk/fera/plants/plant). RPW is Saudi Arabia in 1987 from where it spread to other areas
originally from South and Southeast Asia, where it is a of the country (Abozuhairah et al., 1996; Al-Abdulmohsin,
key pest of coconut, Cocos nucifera, besides infesting 1987). In 1990s it was recorded in several Middle-
other palm species (Nirula, 1956; Wattanapongsiri, 1966; Eastern countries including Iran, Israel, Jordan, the
Li et al., 2009; Matsuura, 1993). Since the mid 1980s the Palestinian Authority Territories, Kuwait and Bahrain
spread of RPW has been rapid mainly through infested (Faghih, 1996; Kehat, 1999). The first report of RPW from

*Corresponding author. E-mail: [email protected]. Tel: (+27) 071 347 6014. Fax: (+27) 015 268 3491.
 El-Shafie et al. 1925

Table 1. Different feeding substrates reported by different authors as constituents of artificial diet for red palm weevil, R. ferrugineus

Feeding substrates Reference

Sugarcane, bagasse, fresh coconut cake, brewer’s yeast, sugarcane, potassium hydroxide, methyl
Rahalkar et al. (1978)
parahydroxyl benzoate, and sorbic acid solutions

Agar, distilled water, brewer’s yeast, wheat germ, corn meal, M-nipagine, benzoic acid, ascorbic acid,
Barranco et al. (1997)
chloramphenicol, coconut fiber, vitamins and amino acids additive and crude protein

Oats, coconut cakes, coconut fruit pieces, vegetable oil, potatoes, soybean flours, date palm leaves, palm
Kaakeh et al. (2001)
fiber sheath, sugarcane fibers, bacto-agar, multi-vitamins, preservatives and water.

Potato, carrot, casein, agar, cereals, and vitamin B and D El-Sebey et al. ( 2003)

Oats, potato, pineapple, and palm fiber sheath Kaakeh (2005)

Agar, brewer’s yeast, wheat germ, corn flour, ascorbic acid, benzoic acid, amino acid-vitamin mix, Martin and Cabello
chloramphenicol and nipagin (2006)

Africa came from Egypt in 1992 (Cox, 1993) while in eradicated (Faleiro, 2006). Pheromone (Ferrugineol)
Europe it was detected in southern Spain in 1995 traps have been used to monitor and mass trap RPW in
(Barranco et al., 1996). The first report of RPW in the area-wide integrated pest management (IPM) programs.
American continent is from the Caribbean Island of However, these conventional food baited pheromone
Curacao (Dutch Antilles) in 2008 where the pest arrived traps (FBPTs) have to be periodically serviced (change
through date palms imported from Egypt for ornamental food bait and insecticide solution) which is labor
purposes http://www.redpalmweevil.com/newlook/RPW intensive. Therefore, a bait-free method to ‘attract and kill’
Report/Caribbean.htm. In USA RPW invasion occurred in RPW adults, has recently been developed for weevil
late 2010 in Laguna Beach, California, USA control in date palm (ElShafie et al., 2011).
(http://www.cdfa.ca.gov/egov/Press_Release ; NAPPO, The term artificial diet means a diet that does not
2010), where it was found infesting Phoenix canariensis include the insect’s natural food except in minute traces
hort. ex. Chabaud. However, morphological parameters and it is often conveniently divided into meridic and
may suggest that this invasion could be the related Asian holidic diets. In the holidic diet all the ingredients, with the
palm weevil, Rhynchophorus vulneratus and not R. common exception of agar, are more or less chemically
ferrugineus. These Rhynchophorus species of weevils defined. In the meridic diet, one or more of the
were synonymised as colour morphs by Hallett et al. ingredients cannot be defined chemically for example,
(2004). RPW is now present in almost all countries of the yeast or wheat germ (Mckinley, 1971). Ideal diet for
Gulf region of the Middle East where it has been insect mass rearing programs should supply all nutrients
designated as a category-I pest of date palm, Phoenix needed to produce acceptable insects, and should be
dactylifera L., by the Food and Agriculture Organization easy to prepare and store for long period, inexpensive
of the United Nations. Economic losses between 1 and and should produce an average yield of adults of at least
5% due to eradication of severely infested palms are 75% from initial viable eggs (Singh, 1983). Various
estimated to range from $5.18 to $25.92 million, feeding substrates were used by many investigators to
respectively (El-Sabea et al., 2009). In the Mediterranean rear RPW in the laboratory for experimental purposes
basin RPW has become a key pest of P. canariensis (Table 1). In many cases, the laboratory rearing and
which is extremely sensitive to its attack (Dembilio et al., mass production of RPW is very important to maintain
2009). Damage to date palm is mainly caused by the purity, age, physiological stage homogeneity and sex-
larval stage feeding within the trunk of palms. Adult based selection to carry out laboratory, semi-field and
females of RPW lay eggs in protected parts of the date field experiments. This improves understanding the life-
palm tree, including wounds on the trunk of established history parameters of RPW including development,
trees, at the base of fronds, the crown of the tree and survivorship and reproduction which will eventually lead
adjacent to offshoots. Early stages of attack are difficult to improvement of control strategies. The ability to
to detect in palms but can be cured with insecticide (stem produce quality insects in large quantity is critical for
injection). Palms in the later stages of attack often harbor many research projects. Rearing of RPW on meridic diet
several overlapping life stages of the pest with extensive could be an alternative to rearing on date palm tissues
tissue damage due to feeding by grubs, and have to be which is expensive and also difficult to follow the
1926 Sci. Res. Essays

Table 2. The different ingredients of the meridic diet used in bioassay experiments.
rearing the red palm weevil, Rhynchophorus ferrugineus 2. Assurance of continuous supply of experimental
insects throughout the year.
Component Amount
Distilled water 875 ml
Agar 18.5 g MATERIALS AND METHODS
Brewer's yeast 45 g
Wheat grains 45 g The meridic diet
Corn flour 45 g
Sorbic acid 1.6 g The larval diet used for mass rearing of RPW was modified from
that developed by Martin and Cabello (2006). The amino acid
Ascorbic acid 4g
mixture used was replaced in our diet with yeast (Saccharomyces
Aminobenzoic acid 1.6 g cerevisiae Meyen ex. E.C. Hansen) produced from date syrup at
®
Pharmaton capsule (1.55g/capsule) 2 capsules DPRC. The main ingredients of our diet included agar,
Tetracycline (250 mg) 2 capsules commercially available baker’s yeast as well as produced from date
syrup at DPRC, wheat meal, corn flour, in addition to other basic
DPRC produced yeast (Saccharomyces cerevisiae) 25 ml ingredients (Table 2). The DPRC produced yeast is rich in amino
and fatty acids and substituted for the commercially available amino
acid mixture as recommended by Martin and Cabello (2006). The
amino acids and fatty acids profiles of the DPRC produced yeast
Table 3. Percentage of amino and fatty acids in the DPRC yeast, are shown in Table 3. PharmatonTM capsules were added to diet to
Saccharomyces cerevisiae used in the meridic diet for rearing of provide vitamins and minerals (Table 4). PharmatonTM is
Rhynchophorus ferrugineus. manufactured by Ginsana SA, Bioggio, Switzerland on behalf of
Boehringer Ingelheim international GmbH Germany. Tetracycline
Amino acid % Fatty acid % 250 mg, produced by Julphar, Gulf pharmaceutical industries, Ras
Al Khaimah, U.A.E. was also incorporated as an antimicrobial
Aspartic 0.40 Caprylic 0.010
agent.
Threonine 0.47 Capric 0.025
Serine 0.26 Lauric 0.175
Glutamic 0.79 Myristic 0.640 Baker's yeasts
Glycine 0.26 Myristoleic 0.140
Alanine 0.58 Pentadecanoic 0.065 The Baker's yeasts used in this study were a commercial Saf-
Valine 0.57 Palmitic 11.100 instant active dry Baker's yeast (S. I. Lesaffre 59703 Marcq,
Methionine 0.27 Palmitoleic 35.095 France) and a strain from Specialized Laboratory: A
Saccharomyces cerevisiae strain NCYC 1530 (laboratories of the
Isoleucine 0.26 Margaric 0.110 National Center Britain). The strain NCYC 1530 was provided in a
Leucine 0.49 Stearic 5.560 freeze-dried form and was kept in its original form until use. This
Tyrosine 0.72 Elaidic 0.140 yeast was produced using date syrup as described by Al-Eid et al.
(2009) and Al-jasass et al. (2010) and was used in the meridic diet.
Phenyalanine 0.42 Oleic 43.240
Fats and fatty acid profile in the yeast reported here were
Histidine 0.36 Linoleic 0.850 determined according to the A.O.A.C. (1995) official method No.
Lysine 1.02 Eicosanoic 0.080 996.01, amino acid composition of the proteins according to the
Arginine 0.52 Linolenic 0.135 method No. 994.12 and Tryptophan according to the method No.
988.15.
Tryptophan 0.09 Docosanoic 0.150
Cysteine 0.07 Tetracosanoic 0.095
Pentacosanoic 0.140
Diet preparation
Hexacosanoic 0.745
Heptacosanoic 0.070 The basic diet was prepared based on the protocol of Martin and
Octacosanoic 0.050 Cabello, 2006. Thirty seven gram of agar was mixed in 875 ml
distilled water in glass beaker. The mixture was put in a microwave
oven at maximum potency for 8 min and stirred using magnetic
stirrer at 2, 4 and 6 min to dissolve the agar. The mixture was
development of the larvae, since they are concealed removed from the oven to facilitate each stirring and finally removed
at 8 min. The main ingredients were mixed with agar jelly using a
inside the palm tissues. The present study was thus kitchen blender which was previously washed with distilled water
carried out to formulate a meridic diet for mass and surface sterilized with 0.5% sodium hypochlorite. The mixture
production of RPW with the following objectives: was homogenized at high speed for 2 to 3 min and left to cool till
60°C. As the diet cooled, two multi-vitamins (PharmatonTM)
capsules and one tetracycline tablet were added while stirring the
1. Develop a process and quality control system to
media. The diet was then poured into surface sterilized plastic
assure availability of high quality research insects containers while still warm and left to solidify and kept in the
including production of different developmental stages of refrigerator for further use. Before being used, the cups containing
the weevil of comparable physiological status for diets were kept at room temperature to warm up. Small hole was
 El-Shafie et al. 1927

Table 4. Vitamins complex, minerals and trace elements used in the preparation of 1000 g of diet
for rearing Rhynchophorus ferrugineus

Vitamins Amount Minerals and trace elements Amount

Vitamin A 5334 IU Copper 4.0 mg
Vitamin D3 400 IU Selenium 100.0 mg
Vitamin E 20.0 mg Manganese 5.0 mg
Vitamin B1 2.8 mg Iron 20.0 mg
Vitamin B2 3.2 mg Zinc 2.0 mg
Vitamin B6 4.0 mg Calcium 200 mg
Vitamin B12 2.0 mg lecithin 200 mg
Biotin 300 mg Magnesium 20.0 mg
Nicotinamide 36.0 mg
Vitamin C 120.0 mg
Folic acid 0.2 mg

made on the diet surface to facilitate the feeding and burrowing of to each 30 ml plastic cup containing 5 g diet with a fine camel hair
the neonate larvae. Diet cups were covered with a lid vented by brush (n=12). Diet cups with the larvae were checked daily for signs
several small holes. of moult (cast skins and head capsules). As the larvae grew, they
were transferred to larger diet cups as mentioned previously. The
larval instars were determined by the number of moults. Visible
Test insects exuviae including presence of head capsules in the diet were used
as evidence of moulting. The surviving larvae were transferred daily
Adult RPW females and males were captured using insecticide-free into new diet cups after inspection. The development of Larvae was
pheromone (Ferrolure TM) traps deployed by Directorate of followed until they reached pupation. The time periods between
Agriculture in Al-Ahsa, Saudi Arabia. Active weevils were carefully instars as well as the number of moults were recorded.
selected from freshly captured collections, placed in perforated
plastic containers and brought to the entomology laboratory of the
Date Palm Research Center (DPRC). Weevils were sexed based Increase in larval biomass
on the presence of a tuff of fine bristles on the dorsal end of the
rostrum in males and which were absent in females. In order to follow the increase in larval biomass, the mean weight of
larvae (n=24) was recorded every three days starting from the first
day of hatching till the last instar. The absolute increase in body
Collection and incubation of eggs biomass (weight/time) was determined using a sensitive four-digit
electric balance, AL 204 Mettler Toledo manufactured by Mettler-
The collected insects were paired and kept in small plastic Toledo Group.
containers for mating. They were provided with a piece of
sugarcane and left for at least 24 h. The males were removed
thereafter, and the females were observed for egg laying. For Width of larval head capsule
production of maximum number of eggs, the females were put in an
incubator at 27°C for 3 to 4 days. A digital vernier caliper was used to measure the width of larval
The pieces of sugarcane in each container were carefully head capsules. The head capsules of the first and second instars
inspected and peeled to search for eggs which were collected and were measured by using microscopy digital USB Camera
transferred into petri dishes using a fine camel hair brush. Only (OptikamTM) programmed with Optika vision pro 4.1 software as
clean ivory colored and shining eggs were collected. The petri these larval stages were too small to handle with a vernier caliper.
dishes contained cotton pads soaked in 10% sugar solution to Permanent microscopic slides of the first and second instars head
provide nutrients for the newly hatched larvae before being capsules were prepared for the measurements using Canada
transferred to the artificial diet. A 90 mm diameter Watman paper balsam as mounting medium. The postmoult/premoult ratios of
was then put on top of the cotton pad to retain the moisture. The head capsule width in successive larval ecdyses were tested using
eggs were then kept for three days in an incubator at 27°C and Dyar's rule (Klingenberg and Zimmermann, 1992).
50% RH for hatching. The newly hatched larvae were then carefully
transferred to cups containing artificial diet. At the early stage of
larval development, only one larva was placed into each cup to Food-fiber pupation technique
avoid cannibalism. From the 4th instar onwards, 3 to 5 larvae were
put together in one cup. Cups of three different sizes (30, 50 and One of the challenges in the successful rearing of RPW on a
200 ml) were used for small (neonate: 1 to 3rd instar larvae), meridic diet is to get the final instar larvae to pupate. Entomologists
medium (4 to 5th instar larvae) and large (6 to 8th instar larvae) have extensively used sugarcane stem for this purpose. However,
specimens respectively. the diameter of stem is a determining factor in the success of this
process. Sugarcane stem with narrow diameter usually leads to
small size adults in addition to its low efficiency (adult emergence).
Development of immature stages The food-fiber pupation technique as the name implies, consisted of
two layers of date palm fiber and a layer of sugarcane pieces (15
Immediately after hatching, one neonate was carefully transferred cm long), split longitudinally and stacked side by side to make a
 1928 Sci. Res. Essays
asas

scissor was used to separate those cocoons which were firmly

attached to the sugarcane pieces. The collected cocoons were put
each in a plastic cup with four holes on the lid. The cups with
cocoons were kept in an incubator at 27 ± 2 °C and 75 ± 5% R.H. in
complete darkness until adult emergence. The percentage of adult
emergence was then calculated.

Oviposition, egg hatchability, and adult longevity

Upon adult emergence individuals were weighed, sexed and 1

male and 1 female paired (10 pairs) were kept in 400 ml plastic
containers provided with sugarcane pieces. These insects were
used to study oviposition and longevity of RPW reared on the test
diet. Sugarcane pieces were replaced every three days after being
thoroughly examined for eggs which were collected and kept in
gg petri dishes to determine the incubation period, fecundity and
Figure 1. RPW larvae growing on the meridic diet. hatchability. To determine the pre-oviposition period, females were
observed every day after emergence from the pupae until the first
Saas egg was laid. The fecundity was evaluated by the number of eggs
laid during the life span of the female after the first egg laying. Adult
longevity was also recorded. The experiment was continued until
the weevil died.

Rearing of red palm weevil (RPW) on date palm trunk

Concurrently, individuals of RPW were reared on date palm (natural

host) bolts in the laboratory to allow for comparison with those
reared on the test meridic diet. The weevils were reared on the
trunk of 3 to 4 year old Khalas cultivar which is predominant in Al-
Ahsa region of Saudi Arabia. The rearing of RPW on date palm
trunk followed the protocol described by Al-Ayedh (2008).

Data analysis

Results of biological parameters are given as mean ± SD.

Figure 2. Food-fiber pupation technique developed to facilitate Regression and correlation analysis, Chi-square, and t-test were
pupation by Rhynchophorus ferrugineus reared on meridic diet performed whenever necessary using MINITAB statistical
software, version 13.30, Copyright 2000; Minitab Inc.
gg
mat 20 cm wide. The sugarcane pieces were sandwiched between RESULTS
the layers of date palm fiber and wrapped with fine plastic mesh.
The sugarcane-date palm fiber mat was rolled into a cylindrical
bundle resembling date palm offshoot (Figures 1 and 2). The date Table 5 shows different biological parameters of RPW
palm fiber and the sugarcane there by simultaneously provided reared on the meridic diet. The Pre-oviposition period
material for the construction of the cocoon for fully matured larvae lapsed about 4.8 day while the incubation period was
that wanted to pupate and nutrients for those larvae that wanted to 3.66 day. The total number of eggs laid per female
feed and require additional time to complete their development. The (fecundity) was on average 198 eggs with an egg
sugarcane stem also provide a solid support that is required by the
larvae during the cocoon spinning process. The food-fiber bundle hatchability or viability of 88.10%. The larval weight of the
was placed in 30 × 20 × 14 cm plastic container specially first instar was 1.7 mg compared with 3.93 g for the last
fabricated with 20 holes (5 mm diameter; 8 on the lid, 4 at the instar (Figure 3). Thus, the biomass increase of the last
bottom and 2 on each side below the container rim) to provide instar relative to the first one was 2311 times. Pupal
aeration and to conserve the required humidity. The plastic period was 33.66 day with a pupation per cent of more
container was kept in a ventilated glass cupboard and was checked
than 97% and adult emergence of 93%. The total life
after three weeks for cocoons and thereafter, regular inspection
was carried every three days. The inspection process was done cycle (egg to adult) was completed in about 82.4 day.
carefully to avoid disturbing the pupating larvae. The efficiency of The average larval period was 45 day while the average
the technique was calculated and expressed as percentage of weight of last larval instar was 3.42 g. The mean
mature larvae that pupated. longevity of the female was 74.8 day while that of the
male was 83.3 day. The mean adult life weight was 0.54
Percentage of adult emergence
g for the male and 0.57 g for the female. There was no
significant difference between the sexes with respect to
Cocoons were carefully collected by hand. Whenever needed, life weight (t= 1.049, df = 1, p = 0.05).
 El-Shafie et al. 1929

Table 5. Important biological parameters of Rhynchophorus ferrugineus reared on meridic diet and
date palm trunk under laboratory conditions.

Mean ± SD
Sl. No. Biological parameter
Meridic diet Date palm trunk
1 Pre-oviposition period (days) 4.80 ± 1.03 (10) 3.80 ± 0.78 (10)
2 Incubation period (days) 3.66 ± 0.88 (12) 3.08 ± 0.79 (12)
3 Fecundity (number of total eggs/female) 198 ± 24.00 (10) 206.30 ± 14.64 (10)
4 Hatchability or viability of eggs (%) 88.10 ± 5.17 (10) 93.10 ± 2.72 (10)
5 Total larval period (days) 45.09 ± 1.44 (22) 36.06 ± 3.24 (20)
6 Number of larval instars 8 ± 0.00 (25) ___
7 Total pupal period (days) 33.66 ± 2.30 (12) 22.5 0 ± 1.78 (12)
8 Total life cycle from egg to adult (days) 82.41 ± 1.54 (10) 61.64 ± 1.93 (10)
9 Weight of last larval instar (g) 3.42 ± 0.21 (10) 5.31 ± 0.32 (10)
10 Head capsule width of last instar (mm) 5.43 ± 0.27 7.42 ± 0.12 (10)
11 Adult male longevity (days) 83.30 ± 1.63 (10) 91.58 ± 2.23 (10)
12 Adult female longevity (days) 74.80 ± 2.20 (10) 81.80 ± 3.55 (10)
13 Adult male life weight (g) 0.54 ± 0.08 (16) 1.02 ± 0.08 (16)
14 Adult female life weight (g) 0.57 ± 0.11 (16) 1.16 ± 0.04 (16)
Figures in parentheses are number of individuals tested.

Table 6. Chi-square comparison (observed and expected) of larval investigation, the amino acids were incorporated in the
head capsule width in Rhynchophorus ferrugineus reared on diet through the yeast (Saccharomyces cerevisiae)
meridic diet
produced from date at DPRC. This yeast was mainly
considered as a protein source, it also contains
Head capsule width (mm)
Instar carbohydrates, lipids, minerals and vitamin B-complex
1 2 3 4 5 6 7 8 (Robinson, 1986; Hamad, 1986). The yeast is essential
Observed 0.68 1.05 1.5 2.0 2.98 3.2 3.9 5.4 component in any artificial diet and can make it either
Predicted 0.82 1.07 1.40 1.82 2.36 3.0 4.0 5.4 successful or non-successful (Cohen, 2003). Eight larval
Dyar’s ratio = 1.3. Chi-square statistic was not significant (χ = 0.08, df= instars were recorded on the meridic during our study
1, p=0.05) indicating that there was no significant difference between and it was technically difficult to trace the number in the
the observed and predicted head capsule measurements. date palm trunk. However, the life cycle of the weevil was
completed in about 62 d on date palm trunk compared
with 82 day on the meridic diet. The shorter life cycle on
This study revealed that in all there were 8 larval date palm could indicate less number of instars. This is in
instars (Table 6). Linear regression analysis showed line with the findings of Ju et al. (2010) who stated that on
significant relationship between larval instars and head suitable host plants, larvae may have fewer instars and
2
capsule width (R =0.978; Figure 4). Plotting the logarithm thus its development can be shortened. The number and
of the head capsule width measurements against the nature of molts can be frequently altered by external
respective instar generated a straight line indicating the factors, mainly temperature, diet and their interaction
conformity to the law. Therefore, head capsule width (Wigglesworth, 1954; Stamp, 1990) and accordingly the
could be used for estimation of larval instar in the number of instars will also be changed. Large
laboratory population of the RPW. A food-fiber pupation discrepancies occur in the literature concerning the
technique was developed in this study with 100% number of larval instars of RPW reared on different diets
pupation efficiency (Figure 1). and under different laboratory conditions. Ju et al. (2010)
reported eight instars on Canary Island (Phoenix
canariensis) and Washington palms (Washingtonia sp).
DISCUSSION They also reported nine instars on Chusan palm
(Trachycarpus fortune), Pindo palm (Butia capitata), and
The suitability of an artificial diet can be compared with silver date palm (Phoenix sylvestris). Seven instars on
respect to development, survival, longevity and fecundity sugarcane and nine instars on coconut palm (Cocos
(Ki-Sang and Lee, 2004). Artificial diets for rearing RPW nucifera) were recorded in Philippines by Jaya et al.
previously reported had the amino acid mixture as a vital (2000) and Viado and Bigornia (1949) respectively. Nilura
component of the diet (Barranco et al., 1997). In this (1956) reported only three instars on coconut in India
1930 Sci. Res. Essays

Figure 3. Increase in biomass during larval development of Rhynchophorus ferrugineus

reared on meridic diet and date palm trunk

0,75
Log (Head capsule width)

LogeY= 0.95X-0.001
R2= 0.978
0,50 P0.05

0,25

0,00

1 2 3 4 5 6 7 8

Instar

Figure 4. Relationship between head capsule width and instar of Rhynchophorus ferrugineus
reared on meridic diet.

while thirteen instars in Canary Island palm (Phoenix generations of economically important insects entirely on
canariensis) were recently estimated by Dembilio and an artificial diet, in many cases there is loss of both
Jacas (2011). This variation in the number of instars fitness and reproductive potential which cause longer
could be attributed to the rearing conditions and development times and lower fecundity (Coudron et al.,
diet/rearing media used by previous workers. Dyar's law 2002). In the present study, RPW was successfully
has been reported to be applicable to other insects by reared on the meridic diet for one generation without any
several workers where head capsule width was used to apparent deformity or difficulty in rearing as compared
determine the number of instars (Klingenberg and with individual insects reared on date palm as natural
Zimmermann, 1992). Differences in estimates of life host. Thus, the meridic diet employed in this work can be
parameters of RPW reared on meridic diet and date palm a suitable substrate for mass rearing of RPW in the
as natural host may have been due to food-type, and laboratory. However, further evaluation is required to
rearing conditions (Joem and Behmer, 1997). Although determine the effect of the test diet on the weevil survival
there is some success in efforts to rear successive and biological parameters over more than one generation
 El-Shafie et al. 1931

as compare to individual insects reared on date palm as Curculionidae) on artificial diet. Egyptian J. Agric. Res. 81:551-554.
Faghih AA (1996). The biology of red palm weevil, Rhynchophorus
a natural host.
ferrugineus Oliv. (Coleoptera, Curculionidae) in Savaran region
(Sistan province, Iran). Appl. Entomol. Phytopathol. 63:16-18.
Faleiro JR (2006). A review of the issues and management of red palm
ACKNOWLEDGMENTS weevil Rhynchophorus ferrugineus (Coleoptera: Rhynchophoridae) in
coconut and date palm during the last one hundred years. Int. J.
Trop. Insect Sci. 26(3):135-154.
The authors wish to thank Prof J. A. Jacas, Departament Hallett RH, Crespi BJ, Borden JH (2004). Synonymy of Rhynchophorus
de Ciències Agràries i del Medi Natural; Unitat Associada ferrugineus (Olivier), 1790 and R. vulneratus (Panzer), 1798
d’Entomologia Agrícola UJI-IVIA;Universitat Jaume I (Coleoptera, Curculionidae, Rhynchophoridae). J. Nat. Hist.
38(22):2863-2882.
(UJI); Castelló de la Plana , Spain for providing valuable Hamad SH (1986). Screening of yeast associated with food from the
insight on the rearing of RPW on artificial diet and also for Sudan and their possible use for single cell protein and ethanol
critically reviewing this manuscript. production. Ph.D. thesis, TU-Berlin, Germany.
The authors acknowledge with thanks the assistance Jaya S, Suresh T, Sobhitha-Rani RR, Sreekumar S (2000). Evidences
of seven larval instars in the red palm weevil, Rhynchophorus
provided by Ibrahim A. Bou-Khowa, Abul-Aziz Al-Mutari,
ferrugineus Oliv. reared on sugarcane. J. Entomol. Res. 24:27-31.
Magid abdel-Aziz and Waleed Alsinien with thanks in Joem A, Behmer ST (1997). Importance of dietary nitrogen and
laboratory rearing of the weevil. The financial support carbohydrates to survival, growth and reproduction in adult of the
provided by Saudi Aramco, through the Saudi Aramco- grasshopper, Ageneotettix deorum (Orthoptera: Acrididae).
Oecologia Berlin 112(2):201-208.
King Faisal University project on the red palm weevil, is
Ju RT, Wang F, wan FH (2010). Effect of host plant on development
appreciated and sincerely acknowledged. and reproduction of Rhynchophorus ferrugineus (Olivier) (Coleoptera:
Curculionidae). J. Pest Sci. DOI 10.1007/s10340-010-0323-4,
Springer-Verlag 2010.
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