Biologia, Bratislava, 62/6: 774780, 2007

Section Cellular and Molecular Biology

DOI: 10.2478/s11756-007-0147-8

Total lipid and fatty acid compositions of Lertha sheppardi

(Neuroptera: Nemopteridae) during its main life stages

Ozlem Cakmak1, Mehmet Bashan2 & Ali Satar2

1
Department of Biology, Education Faculty, Dicle University, 21280 Diyarbakir, Turkey; email: ocakmak@dicle.edu.tr
2
Department of Biology, Science and Art Faculty, Dicle University, 21280 Diyarbakir, Turkey

Abstract: Total lipid and the fatty acid compositions of phospholipid and triacylglycerol fractions, prepared from eggs,
3rd instars of larvae, pupae, male and female adults of Lertha sheppardi, were analyzed by gas chromatography and gas
chromatography-mass spectrometry. The eect of diet (adults nutrition) on fatty acid composition of L. sheppardi adults
was also investigated. Total lipid of L. sheppardi considerably increased in adults compared with immature stages. There
was a signicant decrease in total lipid level in larval stage in contrast with egg stage. Qualitative analysis revealed the
presence of 14 fatty acids during all stages. The major components were C16 and C18 saturated and unsaturated components
which are ubiquitous to most animal species. In addition to these components, one odd-chain (C17:0) and prostaglandin
precursor fatty acids were found. The fatty acid proles of phospholipids and triacylglycerols were substantially dierent. In
phospholipid fraction, monounsaturated fatty acids were the major proportion of fatty acids in both sex of adults and pupae,
whereas polyunsaturated fatty acids were the most dominant fatty acids in eggs and 3rd instars. Results of triacylglycerol
fraction revealed that fatty acid composition of eggs had higher level of C16:1, C18:0 and C18:3n-3 content than that of 3rd
instars and pupae, which suggests accumulation of energetic and structural reserve materials during embryonic development.
At more advanced developmental stages, mainly in adult females, the amount of C16:1 increased once again, which may be
related to the need for accumulation of sucient energy and of carbon reservoir in the developing new vitellum. Percentages
of C18:1 were signicantly high in adult stages compared to other stages. These ndings indicate that the accumulation and
consumption of fatty acids uctuate through dierent development stages. Diet did not eect the fatty acid composition of
L. sheppardi adults.
Key words: developmental stage; fatty acid composition; Lertha sheppardi.
Abbreviations: FA, fatty acid; FAME, fatty acid methyl ester; MUFA, monounsaturated fatty acid; PL, phospholipid;
PUFA, polyunsaturated fatty acid; SFA, saturated fatty acid; TG, triacylglycerol; TLC, thin-layer chromatography.

Introduction et al. 1974), Dendroctonus frontalis (Hodges & Barras

Fatty acids (FAs) carry out various functions in insects.
They are the primary energy source during the periods
of non-feeding, such as diapause and long migratory
ights (Beenakkers et al. 1985), and during the non-
feeding stages of development (Downer 1985). FAs serve
as precursors in the biosynthesis of pheromones, waxes,
and eicosanoids. Moreover, FAs are structural compo-
nents of membranes and defensive secretions (Stanley-
Samuelson et al. 1988), and they are essential compo-
nents in the function of the cuticle (Blomquist & Dill-
with 1985). Because the relative importance of these
functions varies throughout the development, the rate
of FA biosynthesis also would be expected to uctuate
according to physiological needs.
FA proles have been documented and compiled
for a large number of insects, but the changes in FA
composition during development have been studied in
only a few species Periplaneta americana (Kinsella
1966), Heliothis zea (Lambremont 1971), Lyctus plani-
collis (Maulding et al. 1971), Dacus oleae (Madariaga
1974), Galleria mellonella (Janda 1975), Culex pipiens
quinquefasciatus (Albrecht et al. 1977), Ceratitis capi-
tata (Pagani et al. 1980), Acheta domesticus (Cripps &
De Renobales 1988), Manduca sexta (Ogg & Stanley-
Samuelson 1992), Eurygaster integriceps (Bashan et al.
2002) and Melanogryllus desertus (Bozkus 2003) most
of which show some developmental dierences. Parasad
et al. (1986) suggested that the signicant changes
in lipid composition and in the physical properties of
lipophorin occur during metamorphosis from larvae to
pupa in M. sexta. Thus, at the end of the 5th instar,
and at the initiation of the pre-pupa stage, there is a
decrease in lipid content, which is followed by a large
increase 12 h later. According to Akpnar et al. (2003),
there were no dierences between FA proles of eggs,
nymphs, male and female adults of Gryllus campestris.
Miller & Blankenhip (1973) found that larvae and
adult of Vitula edmandsae serratilineela contained rela-
tively high levels of C16:0 (palmitic acid), C16:1 (palmi-
toleic acid) and C18:1 (oleic acid) even when these acids
were not predominant in the dietary lipid.

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Fatty acid analysis of Lertha sheppardi
Neuroptera order is greatly underrepresented in
the literature on insect and lipid biochemistry. Very few
studies on the total FA composition of the Neuroptera
order have been performed (Fast 1970; Lemesle et al.
1997; Nelson et al. 2003). To the best of our knowl-
edge, nobody has investigated FA composition of phos-
pholipid (PL) and triacylglycerol (TG) fractions of an
insect belonging to this order.
Nemopteridae are unique insects by virtue of their
long lamentous or ribbon-like hind wings, which often
have broad dilations. Adults feed exclusively on pollen
and nectar facilitated by long specialized mouth parts,
whilst all larvae are carnivores (Mansell 1992). Lertha
sheppardi is endemic to Turkey (Satar & Ozbay 2004).
The aim of this study was to investigate the
changes in PL and TG FA compositions of L. shep-
pardi males and females in various development stages
of the insect such as eggs, 3rd instars, and pupae. We
also wanted to evaluate the eect of diet on FA compo-
sition of female adults. The results are compared with
reported changes in FA composition and with physio-
logical and biochemical events.
Material and methods
Insects
Eggs of L. sheppardi were obtained from gravid females col-
lected at dierent points of South East of Turkey (2004
2005). Gravid females were individually separated and
placed individually into plastic jars in order to obtain eggs
under laboratory conditions. Some drops of water, a few
grains of commercial pollen (Aksuvital), and fresh Lisea
strigosa with owers (belonging to the Apiaceae) were in-
troduced into each jar. Once laid, eggs were transferred
into small vials for larval development. The larvae were
kept in laboratory, using the methods of Monserrat & Mar-
tinez (1995). Larvae of L. sheppardi were fed on Tenebri-
onids, Tipulids and small arthropods in laboratory condi-
tions. Eggs, 3rd instars, pupae and adults were transferred
into chloroform/methanol (2:1, v/v). They were kept deep
frozen (20 C) until use. Adults of L. sheppardi feed on
owers ofL. strigosa and these tissues were prepared for
analysis.
Analysis of total FA of L. strigosa
Total lipids were extracted from three groups of 5 g of fresh
owers of L. strigosa by the method of Bligh & Dyer (1959).
Autoxidation of unsaturated FAs was minimized by addi-
tion of 50 L of 2% butylated hydroxytoluene in chloroform
to each sample. Saponication preparation of FA methyl
esters for gas chromatography was performed as described
(Stanley-Samuelson & Dadd 1981). The extracts were dried
under N2 and then taken up in 1.5 mL of 1.0 N KOH and
1.5 mL of methanol. The tubes were sealed under N2 then
reuxed at 85 C for 2 h. Non-saponiable lipids were sep- the FAMEs were determined by analysis of spectra and by
arated by adding 3 mL of water and then extracting with
5 mL of petroleum ether. The FA-containing aqueous layer
was acidied by addition of concentrated sulfuric acid. FAs
were then extracted three times with petroleum ether. The
samples were dried under N2 and FA methyl esters were
formed by reuxing in acidied methanol for 1 h. The tubes
were cooled and after adding 3 mL of water, the methyl es-
ters were extracted three times with 3 mL petroleum ether.
775
Analysis of insects FAs
The insects were processed for lipid extraction and analy-
sis following the methods described by Howard & Stanley-
Samuelson (1990). Total lipids were extracted by homog-
enizing groups of 30 3rd (last) instars, 25 pupae, 20
adult males, and 20 adults females per sample in chlo-
roform/methanol (2:1, v/v). Autoxidation of unsaturated
components was minimized by adding 50 L of 2% buty-
lated hydroxytoluene in chloroform to each sample during
the extraction process.
The total lipid extracts were dried under a stream of
N2 , then PL and TG fractions were isolated by thin-layer
chromatography (TLC), using Silica Gel G TLC plates (20
20 cm, 0.25-mm thick). After applying the total lipid ex-
tracts, the TLC plates were developed in petroleum ether:
diethyl ether: acetic acid (80:20:1, v/v). Lipid fractions
were made visible by spraying the TLC plates with 2,7-
dichlorouorescein (Supelco, Supelco Park, PA, USA), and
PL and TG fractions were identied by corresponding stan-
dards.
The PL and TG fractions were scraped into reaction
vials, and the associated FAs were transmethylated by re-
uxing the fractions in acidied methanol for 90 min at
85 C. The fatty acid methyl esters (FAMEs) were extracted
from the reaction vials three times with hexane, and con-
centrated.
Total lipids were conducted using the method of
Christie (1982).
Gas chromatography
The FAMEs were analyzed on gas chromatography using
an Ati Unicam 610 gas chromatograph equipped with a
SP-2330 capillary column (0.25 mmX 30 M, 0.2 m lm
thickness, Supelco), a ame ionization detector and an Uni-
cam 4815 recording integrator. The separations were con-
ducted with temperature programming from 180 to 200 C
at 5 C/min, after an initial 2min hold. FAMEs were identi-
ed by comparisons of retention times with authentic stan-
dards (Sigma Chemical Co., St. Louis, MO, USA). Individ-
ual FAMEs were identied by comparisons with the chro-
matographic behaviors of authentic standards.
Gas chromatography-mass spectrometry
The chemical structures of the FAMEs were conrmed by
capillary gas chromatography-mass spectrometry (GC-MS).
GC-MS analyses were made using a GC-MS equipment (HP
5890E series GC-System, Hewlett-Packard, Palo Alto, CA,
USA) with mass-selective detection. An Innowax column (30
m 0.25 mm i.d., 0.25 m lm thickness) was used, and
the temperature was programmed from 150 to 230 C at a
2 C/min increase with an initial hold of 36 min. The carrier
gas was helium (1 mL/min) and the split ratio was 1:50. The
injection port and the detector temperatures were 250 and
300 C, respectively. The mass spectrometer was operated
in the electron impact ionization mode (70 eV). Chemical
structures of the FAMEs were identied by comparison with
the Wiley 275 and Nist 98 databank. Chemical structures of
comparing obtained spectra with the spectra of authentic
standards.
Statistical analysis
Statistical analyses of percentages of FAs were tested by
analysis of variance (ANOVA) and comparisons between
means were performed with Tukey test. Dierences between
means were evaluated as signicant if P < 0.05.
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776 O. Cakmak et al.

Table 1. The percentage of total lipid of L. sheppardi at dierent
main life stages.
Main life stages Total lipid (%)
Eggs 15.38
3rd instars 8.60
Pupa 9.91
Female adult 13.77
Male adult 14.49
Results
The percentage of total lipid of L. sheppardi at dier-
ent development stages and sexes are shown in Table 1.
They were relatively high in eggs, but they sharply de-
creased in the 3rd instars. Total lipid content increased
in adults, especially in males.
The FA compositions of PLs and TGs prepared
from eggs, 3rd instars, pupa, males and females of L.
sheppardi are given in Tables 2 and 3. There were dif-
ferences between the lipid fractions. The major FAs
comprising the PL fraction were C18:1, C16:0, and
C18:2n-6 (linoleic acid). Small amounts of C12:0 (lau-
ric acid), C14:0 (myristic acid), C17:0 (heptadecanoic
acid), C20:1 (eicosenoic acid), and C20 PUFAs were
also observed in these fractions (Table 2). The TGs
are composed primarily of C16:0, C18:0 (stearic acid),
and C18:1. In addition to these components, quanti-
tatively minor FAs were present (Table 3). Odd-chain
FA and eicosanoid precursor C20 PUFAs were detected
by ame ionization gas chromatography only sporadi-
cally; they were in low titres. More sensitive analysis by
GC-MS conrmed that these components are present
in both fractions. Especially the PLs were higher than
TGs in C20 PUFAs.
There were uctuations in level of some FAs
at dierent development stages. In PL fraction, the
major saturated FAs of eggs were C16:0 and C18:0
(12.80% and 8.50%, respectively). The major unsatu-
rated FAs in eggs were C18:1 and C 18:2n-6 (31.25%
and 30.05%, respectively). Low proportions of C14:0,
C20:3n-6 (eicosatrienoic acid) and C20:4n-6 (arachi-
donic acid) were detected in eggs and all other main
life stages. However, C17:0 was detected only in adults
and pupae. The content of C18:2 showed signicant de-
crease starting from the 3th instars, but showed an in-
crease in adults. There were no important dierences
in both sexes, except C16:0 and C18:0.
All of the three classes of FAs, the saturated (SFAs;
F = 625; P < 0.001), monounsaturates (MUFAs; F =
1954; P < 0.001), and polyunsaturates (PUFAs; F =
1153; P < 0.001), varied signicantly through main life
stages. MUFAs constituted the major proportion of FAs
in both sexes of adults and pupae, whereas PUFAs were
the most dominant FAs in eggs and 3rd instars. The
proportion of SFAs was always the lowest in each stages
or sex, but they considerably increased in abundance in
3rd instars and pupae in comparison to that of egg or
adult stages.
In TG fraction, the FA proles from eggs and 3rd
instars were not the same; there was a decrease in MU-
FAs (Table 3). In addition, eggs diered from the other
stages with lower proportions of C18:2 and higher pro-
portions of C18:0 and C18:3n-3 acids. Percentage levels
of C18:1 were shown signicantly high in adult stages
in contrast to other stages. In this fraction, the pro-
portions of SFAs increased more signicantly than PL
fraction.
The FA compositions of PLs and TGs prepared
from female adults, and the total FA composition of
L. strigosa on which the adults were fed are presented
in Table 4. We selected female adults because there
were no signicant dierences between male and fe-

Table 2. FA compositions, as proportions of total FAs, in phospholipids prepared from total lipid extract of whole L. sheppardi at the
indicated main life stages.a

Fatty acid Eggs 3rd instars Pupae Adult females Adult males

C12:0 0.50 0.08a 0.73 0.06a 0.75 0.07a 1.02 0.11b

C14:0 0.31 0.05a 0.20 0.08b 0.19 0.09b 0.36 0.09a 0.67 0.03c
C16:0 12.80 1.51a 15.47 0.80b 13.01 0.97a 16.19 3.04b 13.02 1.72a
C16:1 4.93 1.04a 2.33 0.41b 4.10 0.59a 4.81 0.53a 4.14 0.54a
C17:0 0.11 0.06a 0.30 0.07a 0.57 0.09a
C18:0 8.50 1.10a 11.18 0.65b 13.10 1.09c 6.53 0.54d 9.05 0.93a
C18:1 31.25 2.54a 25.64 2.26b 36.08 2.46c 38.35 3.15d 39.03 2.84d
C18:2n-6 30.05 2.81a 38.13 3.20b 27.02 2.12c 28.10 1.16c 29.02 1.03c
C18:3n-3 4.31 0.33a 2.89 0.14b 3.09 0.77b 2.59 0.20b 1.63 0.45b
C20:1n-9 1.03 0.07
C20:2n-6 0.19 0.06a 0.03 0.08b 0.20 0.06a
C20:3n-6 2.56 0.71a 2.10 0.77b 1.50 0.12c 0.50 0.31d 0.70 0.23d
C20:4n-6 2.50 0.27a 1.30 0.12b 1.60 0.57b 1.12 0.30b 1.29 0.33b
C20:5n-3 0.06 0.03a 0.40 0.08b 0.22 0.23b

SFA 23.12 2.33a 27.58 2.11b 26.41 1.65b 24.13 2.60c 24.31 1.02c
MUFA 37.21 3.47a 27.97 2.75b 40.18 3.77c 43.16 3.65d 43.19 3.50d
PUFA 39.48 2.13a 44.45 3.08b 33.41 1.94c 32.69 3.23d 32.87 3.08d

a The values are shown as mean SD. Means are the averages of three replicates using 30 3rd instars, 25 pupae, 20 adult males, and
20 adult females per replicates. Means with the same letter in each row do not signicantly dierent from each other, P > 0.05.

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Fatty acid analysis of Lertha sheppardi 777
Table 3. FA compositions, as proportions of total FAs, in TGs prepared from total lipid extract of whole L. sheppardi at the indicated
main life stages.a

Fatty acid Eggs 3rd instars Pupae Adult females Adult males

C12:0 0.15 0.02a 0.44 0.05b 0.23 0.04a 1.02 0.07c 0.70 0.11c
C14:0 3.75 0.13a 4.58 0.05b 1.02 0.11c 0.74 0.02d 2.08 0.02e
C16:0 29.23 2.47a 31.02 3.85b 34.94 1.80c 23.03 3.07d 19.90 1.69e
C16:1 2.32 0.22a 1.10 0.91b 0.90 0.04b 3.82 0.33c 5.63 0.42d
C17:0 0.15 0.08a 1.02 0.02b 1.15 0.14b
C18:0 35.83 2.14a 33.18 2.77b 26.95 1.09c 13.82 0.63d 14.36 0.22d
C18:1 20.50 2.40a 16.13 1.61b 24.28 2.54c 45.61 3.82d 45.56 3.66d
C18:2n-6 5.20 1.01a 12.68 1.28b 10.94 1.14b 7.43 0.36c 8.73 1.09c
C18:3n-3 4.31 0.33a 2.89 0.14b 3.09 0.77b 2.47 0.20b 1.38 0.45c
C20:1n-9 1.01 0.05
C20:2n-6 0.08 0.01
C20:3n-6 0.20 0.30
C20:4n-6 0.35 0.12a 0.12 0.30b 0.25 0.33c
C20:5n-3 0.08 0.01a 0.11 0.03a

SFA 68.81 4.01a 69.22 3.47a 63.29 3.31b 39.63 2.13c 38.18 2.09d
MUFA 23.83 2.40a 17.23 1.71b 25.18 2.04a 51.54 3.83c 52.84 3.92c
PUFA 7.21 0.94a 13.53 1.28b 11.12 1.57c 8.92 0.88d 9.43 1.05d

a The values are shown as mean SD. Means are the averages of three replicates using 30 3rd instars, 25 pupae, 20 adult males, and
20 adult females per replicates. Means with the same letter in each row do not signicantly dierent from each other, P > 0.05.

Table 4. Proportions of FAs, as percentage of total FAs, in the phospholipid and TG fractions of total lipid extracts from female adults
of L. sheppardi and their respective adult diet.a

Fatty acid Phospholipid Triacylglycerol Total FAs of Diet

C12:0 0.75 0.07a 1.02 0.07a

C14:0 0.36 0.09a 0.74 0.02a 4.01 0.08b
C16:0 16.19 3.04a 23.03 3.07b 27.92 1.19c
C16:1 4.81 0.53a 3.82 0.33b 1.20 0.03c
C17:0 0.30 0.07a 1.02 0.02b 0.42 0.10a
C18:0 6.53 0.54a 13.82 0.63b 1.93 0.23c
C18:1 38.35 3.15a 45.61 3.82b 3.18 0.87c
C18:2n-6 28.10 1.16a 7.43 0.36b 36.63 3.16c
C18:3n-3 2.59 0.20a 2.47 0.20a 24.71 2.07b
C20:3n-6 0.50 0.31
C20:4n-6 1.12 0.30a 0.12 0.30b
C20:5n-3 0.40 0.08a 0.08 0.01b

SFA 24.13 2.60a 39.63 2.13b 34.28 3.31c

MUFA 43.16 3.65a 51.54 3.83b 4.38 2.04c
PUFA 32.69 3.23a 8.92 0.88b 61.34 1.57c

a The values are shown as mean SD. Means are the averages of three replicates using 20 adult females per replicates. Means with

the same letter in each row do not signicantly dierent from each other, P > 0.05.

male adults. The major components in L. strigosa in-
clude C16:0, C18:2n-6, and C18:3n-3. We observed im-
portant dierences between insect FA proles and their
dietary proles. In particular, the L. strigosa had higher
proportions of C18:2n-6 (37% vs. 28%) and C18:3n-3
(25% vs. 2%), but lower proportions of C18:1 (3% vs.
38%) compared to PLs. The same dierences were also
detected in TG fraction. We also note the evidence for
the biosynthesis of C20 PUFAs from their dietary C18
counterparts. The PL fatty acids of female adults in-
cluded 20:3n-6, 20:4n-6, and 20:5n-3 (eicasopentaenoic)
acids, which are characteristically absent from plant
lipids.
Discussion
We observed that the amount of total lipid changed ac-
cording to main life stages and sexes of L. sheppardi.
Results revealed that the total lipid amount is signif-
icantly higher in eggs than that in 3rd instars, which
suggests an accumulation of energetic and structural
reserve materials during vitellogenesis and a late con-
sumption during embryonic development. Several stud-
ies (Cripps et al. 1988; Nestel et al. 2003) have shown
that insect lipids increase because of absorption or syn-
thesis from food material during larval stages. Pupae of
many insects contain large amounts of lipids deposited
during larval life, and these large stores are metabolized
slowly during the pupal adult transformation to provide
energy for new synthesis and cellular changes (Agrell &
Lundquist 1973; Beenakkers et al. 1985). Changes in
lipid stores occur because of ight, mating activities,
and oviposition (Gilby 1965) in the adult stage. In this
study, the total lipid amount increased during the pu-

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778
pal stage relative to the larval stage. However, we did
not observe a signicant increase in lipid content, be-
cause we assayed the last larval stage before pupation
in treatments. The adult stage had higher lipid con-
tent than that of immature stages. The accumulation
of lipid in adults may be related to ight and reproduc-
tive activities. Candy & Kilby (1975) reported that the
signicant increases in lipid content of males relative
to females might be related to higher ight activities
of males in search of females for mating. Also, females
might have laid their eggs, and consumed some of their
lipid storage before extractions were performed. During
ovarial development, lipids synthesized in the fat body
are transported to the developing ovary, and stored for
use in embryogenesis.
The FA compositions during main life stages of
male and females of L. sheppardi were also studied
in this work. The data presented here indicate that
the quantitatively major FAs associated with PLs and
TGs prepared from L. sheppardi include C16:0, C18:0,
C18:1, C18:2n-6 and C18:3n-3, as reported for many
other Neuropthera and most other insect orders (Fast
1970). Although these components are common to most
insects, the FA compositions of various species are
considerably variable. Certain groups of insects stand
out because they exhibit unusual and characteristic
FA proles (Stanley-Samuelson et al. 1988). Dipterans
are characteristically high in C16:1 (Fast 1970). The
dipteran pattern of high C16:1 was also seen in four
heteropterans: Blissus leucopterus leucopterus, Blissus
iowensis (Spike et al. 1991), Eurygaster maura L. (Kil-
incer et al. 1987) and Eurygaster integriceps (Bashan
et al. 2002). However, proportion of C16:1 is low in
most Heteroptera (Thompson 1973; Bashan & Cakmak
2005). Some aphids have up to 80% of C14:0, and coc-
cids are characterized by high proportions of C10:0 and
C12:0 (Fast 1970). The waxmoth Galleria mellonella
diers from other insects with high proportions of C20:1
in TGs of males but not females (Stanley-Samuelson
1984). Aquatic insects stand out with very high propor-
tions (> 20%) of C20:4n-6 and C20:5n-3 components in
their tissue lipids (Hanson et al. 1985). C20 PUFAs were
also detected in Collembola specimens from a decidu-
ous woodland at proportions up to 29.7% of total FA
composition (Chamberlain & Black 2005). We did not
observe unusual FA prole in L. sheppardi unlike the
certain insects.
The FA prole in insects varies considerably. Many
factors inuence the FA composition, such as enviro-
mental temperature, stage of development, diet, sex,
diapause, and migratory ight (Beenakkers et al. 1985).
In particular, development and diet exert strong inu-
ences on the shape of FA proles. This is the case for the
mosquito Culex torsalis where larvae appear to be far
lower than adults in C18:1 (Takata & Harwood 1964).
The FA proles of 20day-old larvae of the beetle Lyc-
tus planicollis are very dierent from those of 60day-
old larvae in terms of C18:1 (Maulding et al. 1971). PL
FA proles dier among eggs, larvae, pupae, and adults
of the pine beetle Dendroctonus frontalis (Hodges &
O. Cakmak et al.
Barras 1974). Ogg & Stanley-Samuelson (1992) showed
that FA proles of PLs and TGs from Manduca sexta
are dierent in each stage of development. Total FA
compositions of developmental stage of Apanteles gal-
leria were also signicantly dierent from each other
(Nurullahoglu et. al. 2004).
The data reported here similarly indicate that the
FA proles of PLs and TGs from whole insects of L.
sheppardi are dierent at each stage of development.
For example, in PL fraction, MUFAs constituted the
major proportion of FAs in both sexes of adults and
pupae, whereas PUFAs were the most dominant FAs
in eggs and 3rd instars. The proportion of SFAs was
always the smallest in each stages or sex, but they con-
siderably increased in abundance in 3rd instars and pu-
pae in comparison with that of egg or adult stages. In
TG fraction, the FA proles from eggs and 3rd instars
were not the same; there was a decrease in the MUFAs.
In addition, eggs diered from the other stages, with
lower proportions of C18:2n-6, and higher proportions
of C18:0 and C18:3n-3 acids. Results of TG fraction
revealed that FA composition of eggs had higher level
of C16:1, C18:0 and C18:3n-3 content than that in 3rd
instars and pupae, which suggests an accumulation of
energetic and structural reserve materials during em-
bryonic development. At more advanced developmen-
tal stages, C16:1 increases once again, mainly in adult
females, which may be related to the need for accumu-
lation of sucient energy and of carbon reservoir in the
developing new vitellum. In general, TG FA proles do
not change through the excursion of the life cycle. How-
ever, in L. sheppardi, the TG FA proles changed. TGs,
the typical lipids of energy reserve, are the major lipids
consumed before eclosion of the eggs, probably to sup-
port the metabolic eort implied in embryogenesis. The
carbon needed in carbohydrate and protein synthesis
during embryogenesis could also be supplied by degra-
dation of TGs. TGs whose major repository is the fat
body in insects are frequently associated with species
that undergo lengthy periods of metabolic activity (e.g.,
diapause or migratory ights) or that have non-feeding
stages of development (e.g., pupae, adult). However,
PLs function as structural components of membranes
(Downer 1985).
The unsaturated FAs to saturated FAs ratio for
PLs of L. sheppardi was signicantly higher than the ra-
tio calculated for TGs, mainly because of the high pro-
portion of C18:1 and C18:2n-6 acids. One of the promi-
nent patterns in insect FA compositions is that PUFAs
are generally found in higher proportions of PLs, as
opposed to neutral lipids (Stanley-Samuelson & Dadd
1983). C18 PUFAs, especially C18:2n-6 acid, tend to
occur preferentially in insect PLs (Howard & Stanley-
Samuelson 1990; Stanley-Samuelson et al. 1990); and
our data are in accord with this general observation, as
C18 PUFAs, C18:2n-6, and C18:3n-3 (linolenic acid),
are found in considerably higher proportions for the
PL FAs than that for the TGs. High levels of these 18
PUFAs in the PL could be based on a number of phys-
iological factors. One of the major functions of C18:2n-
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Fatty acid analysis of Lertha sheppardi
6 acid and PUFAs, in general, is that they serve as a
structural component of membranes to maintain proper
uidity and permeability. C18:2n-6 acid is also a precur-
sor to arachidonic acid (C20:4n-6), a PUFA precursor to
the eicosanoids, including prostaglandins, leukotrienes,
and thromboxanes (Stanley 2006). The FA composi-
tions can be modied by hydrolysis of some FAs, to-
gether selective reacylation of others, and also by chang-
ing the existing components (Stanley-Samuelson et al.
1992).
There are a number of studies which observed the
inuence of dietary lipids on insect lipid composition.
Lambremont et al. (1964) found that the tissue FAs in
adult Anthonomus grandis approximately matched its
diet; however, some FAs, not detected in the diet, were
found in larval and adult insects. Trichopluia ni (cab-
bage looper) larvae can drastically alter the composi-
tion of FAs available in the diet (Nelson & Sukkestad
1968). Heliothis zea larvae reared on a wheat-germ
diet have a very dierent FA composition than those
which are reared on broad-beans (Schaefer 1968). The
TG composition of Apis mellifera appears to be inu-
enced by the diet. The triglycerids of both species of
Diptera (Sarcophaga bullata and Phormia regina) con-
tained large amount of C16:1 at all development stages,
in contrast to the composition of the diet (Harlow et al.
1968). The FA composition of body lipid of the house
cricket, Acheta domesticus, is much the same whether
it is fed on wheat germ oil or not, and males resem-
ble females, except for linolenic acid (McFarlane et al.
1984). Howard et al. (1992) found that six FAs occurred
in signicantly dierent proportion between the labo-
ratory and feral population of Tenebrio molitor.
In this study, a comparison of the total FA com-
position of the diet, and PL and TG FA compositions
of female adults indicate that the FA composition of L.
sheppardi lipids is not identical to the FAs of the di-
etary lipids. In female adults, the major FA was oleic
acid which made up about 38% in PL and 45% in TG
fractions, but this component was only 3% of the di-
etary. We infer that L. sheppardi are able to produce
18:1 de novo, as seen in most animals. L. sheppardi
seem to store 18:1 or utilize two PUFAs, 18:2n-6, and
18:3n-3, preferentially. Since the insect lipids, generally
TGs, are higher and lower respectively in these acids
than is the insect food. Also, adults of L. sheppardi
are able to biosynthesize C20 PUFAs from their C18
counterparts, as reported for a few other insect species
(Stanley- Samuelson et al. 1988). We suggest that L.
sheppardi is able to modify its FA compositions, prob-
ably to suit the physiological requirements.
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Received February 23, 2007
Accepted August 3, 2007
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