Performance of Lymantria xylina (Lepidoptera: Lymantriidae)

on Artificial and Host Plant Diets

Author(s): Tse-chi Shen , Chih-ming Tseng , Li-ching Guan , and Shaw-yhi
Hwang
Source: Journal of Economic Entomology, 99(3):714-721. 2006.
Published By: Entomological Society of America
DOI: http://dx.doi.org/10.1603/0022-0493-99.3.714
URL: http://www.bioone.org/doi/full/10.1603/0022-0493-99.3.714

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 ECOLOGY AND BEHAVIOR

Performance of Lymantria xylina (Lepidoptera: Lymantriidae) on

Artificial and Host Plant Diets
TSE-CHI SHEN, CHIH-MING TSENG, LI-CHING GUAN, AND SHAW-YHI HWANG1
Department of Entomology, National Chung Hsing University, Taichung, Taiwan 402

J. Econ. Entomol. 99(3): 714Ð721 (2006)

ABSTRACT Lymantria xylina Swinhoe (Lepidoptera: Lymantriidae) is a serious defoliator of hard-
wood and fruit trees in Taiwan. The larvae of L. xylina feed on ⬎63 species of host plants, belonging
to 29 families. Because a large number of larvae are needed for the production of nucleopolyhedrosis
virus (NPV) or other related studies, the development of a suitable artiÞcial diet is very important for
the mass rearing of this moth in the laboratory. In this study, eight artiÞcial diets, modiÞed from
different formulas, and one host plant, Liquidambar formosana Hance, were used to feed L. xylina
caterpillars. Through various bioassays (Þrst instar survival trial and long- and short-term feeding
trials), the most suitable diet for the L. xylina was selected by performance comparisons with L.
formosana. After the Þrst instar survival trial, two of the diets were discarded, because no larva survived
on these diets. The results of the long-term feeding trial indicated that the larvae grew successfully
on only three kinds of artiÞcial diet. Finally, results of the short-term feeding trial revealed that a diet
(diet A), modiÞed from the gypsy moth, Lymantria dispar (L.), formula diet, was the most appropriate
for the L. xylina. Larvae fed on diet A had better survival rate, pupal weight, adult size, efÞciency of
conversion, and relative growth rate than larvae fed on other diets; they did not grow as well as those
fed on L. formosana, however, except for pupal and adult weight, and approximate digestibility. In
summary, diet A was found to be the best of the artiÞcial diets for the L. xylina and is suitable for mass
rearing of this moth in the laboratory.

KEY WORDS Lymantria xylina, Liquidambar formosana, artiÞcial diet, feeding trial

Lymantria xylina (Lepidoptera: Lymantriidae) is a
major pest of casuarina, Casuarina equisetifolia L., and
acacia, Acacia confusa Merr., forests, natural areas, and
fruit tree orchards in Taiwan, Japan, India, and the
eastern coast of mainland China (Chao et al. 1996). L.
xylina is closely related to the gypsy moth, Lymantria
dispar (L.), which is an extremely serious forest pest
in North America. Similar to the gypsy moth, L. xylina
is univoltine. Shortly, after emergence and mating in
summer, females lay a single, hair-covered egg mass,
consisting of ⬇100 Ð1000 eggs (Shen et al. 2003). Like
L. dispar, eggs of the L. xylina undergo diapause as
pharate Þrst instars, before the complete consumption
of the extra embryonic yolk (Gray et al. 2001, Hwang
et al. 2004). The larvae hatch in April of the following
year, after an ⬇8 Ð9-mo dormancy (Hwang et al. 2004).
Larval stages last ⬇1.5Ð2 mo, having Þve to eight in-
stars (Chang and Weng 1985, Chao et al. 1996). The
pupal stage lasts ⬇2 wk. Adult males emerge Þrst,
followed several days later by adult females.
L. xylina is a polyphagous herbivore, and the num-
ber of recorded host plants for this moth includes and
most likely exceeds 69 species of trees and shrubs,
belonging to 29 families (Chang and Weng 1985, Chao
1 Corresponding author, e-mail: oleander@dragon.nchu.edu.tw.
et al. 1996). Outbreaks of L. xylina infestations have
only been found in casuarina plantations; however,
serious defoliation on fruit trees also has occurred in
several areas of central Taiwan (Chao et al. 1996).
Current infestations of fruit trees and other hard-
woods by L. xylina indicate the real threat posed by
this moth.
Although a potentially serious pest of hardwood and
fruit trees in Taiwan, little is known about L. xylina
(Chao et al. 1996, Gries et al. 1999, Shen et al. 2003).
For management purposes, more studies must be un-
dertaken to understand its population dynamics, the
relationship with its host plants, and potential con-
trolling agencies. To study this moth, mass production
in the laboratory is necessary. Generally, the food
sources used to rear herbivorous insects in the labo-
ratory can be divided into natural hosts and artiÞcial
diets (Kao 1995). Past studies have indicated that the
larvae of some insect species performed better on
natural host plants than on artiÞcial diets (Hirai 1976,
Hou and Hsiao 1986, Dosdall and Ulmer 2004). How-
ever, for research purposes, problems can arise when
rearing caterpillars on natural host plants. First, plants
are a suboptimal food for insect herbivores, because of
inadequate nutrient ratios and the occurrence of al-
lelochemicals (e.g., tannins and alkaloids) that need to

0022-0493/06/0714Ð0721$04.00/0 䉷 2006 Entomological Society of America

June 2006 SHEN ET AL.: L. xylina ON DIFFERENT DIETS 715

Table 1. Composition of artificial diets for rearing of L. xylina

ArtiÞcial diet
Component Ia IIb IIIc IVd
A B C D E F G H
Wheat germ 23 g 20 g 18 g 15 g 18 g 15 g 12 g 9g
Formosan sweet gum leaf powder 3g 3g 3g 3g
Soybean powder 6g 6g 5g 5g
Kidney bean powder 5g 5g
Corn powder 14.5 g 14.5 g
Yeast powder 5g 5g 3g 3g 3.6 g 3.6 g
Casein 5g 5g
Cholesterol 1g 1g 1g 1g 1g 1g 1g 1g
WessonÕs salt mix. 1.6 g 1.6 g 0.5 g 0.5 g 0.5 g 0.5 g
Ascorbic acid 0.5 g 0.5 g 0.5 g 0.5 g 0.5 g 0.5 g 0.5 g 0.5 g
Vitamin mix. 1.5 g 1.5 g 1.5 g 1.5 g 0.5 g 0.5 g
L-Cysteine 0.1 g 0.1 g
Methyl-p-hydroxybenzoate 0.4 g 0.4 g 0.3 g 0.3 g 0.3 g 0.3 g
Chloramphenicol 0.05 g 0.05 g 0.05 g 0.05 g 0.05 g 0.05 g
Sorbic acid 0.4 g 0.4 g 0.05 g 0.05 g 0.05 g 0.05 g 0.05 g 0.05 g
Agar 3g 3g 3g 3g 3g 3g 3g 3g
Distilled water 180 ml 180 ml 180 ml 180 ml 180 ml 180 ml 180 ml 180 ml

a
ModiÞed from ODell et al. (1985).
b
ModiÞed from Tang (unpublished).
c
ModiÞed from Tang (unpublished).
d
ModiÞed from Kao (1995).

be detoxiÞed (Bailey 1976, Schoonhoven et al. 1998,
Cohen 2004). Second, plant material is difÞcult to
standardize, because individual plants and plant parts
may greatly vary with season, developmental stage,
and so on. However, artiÞcial diet formulations allow
for precise management of nutritional factors, and
they have been developed for many insect species,
beginning in the 1950s (Schoonhoven et al. 1998).
Especially for L. xylina, mass rearing is a requisite in
the production of the microbial insecticide nucleo-
polyhedrosis virus (NPV) or can provide individuals
for other research. No artiÞcial diet, however, has yet
been developed for this moth. Thus, the objective of
this study was to assess the effects of some of the
artiÞcial diets of common lepidopteran pests on the
performance of L. xylina, in the hope of Þnding a
suitable artiÞcial diet for this moth. In addition, we also
compared the performance of L. xylina on artiÞcial
diets and on one of its natural host plants, Liquidambar
formosana Hance.
Materials and Methods
Source and Treatment of Egg Masses. In late May
2003, L. xylina egg masses were collected from two
heavily overrun areas in central Taiwan, according to
Shen et al. (2003). These areas were located at
Mingchien, Nantoa (23⬚ 49.1⬘ N, 120⬚ 39.3⬘ E) and
Erhshui, Changhua (23⬚ 50.0⬘ N, 120⬚ 36.4⬘ E). Ap-
proximately 200 egg masses were collected from each
site.
In the laboratory, the egg masses were surface-
sterilized, by soaking in 0.1% sodium hypochlorite
with 1% Tween 80 solution for 10 min., and rinsing
under running tap water for 5 min. Then, the egg
masses were air-dried. To obtain individual eggs, the
egg masses were broken up and the hairs removed, by
rubbing against sticky tape. The dehaired eggs, from
different egg masses, were mixed and placed into petri
dishes (⬇500 eggs/dish). These dishes were then
moved to a Percival growth chamber (Percival, Perry,
IA) to expose them to chilled conditions (10⬚C), to
terminate diapause (Hwang et al. 2004). After 120 d of
chilling, the eggs were sterilized again and placed in
the Percival growth chamber at 28⬚C:25⬚C (L:D) with
a photoperiod of 12:12 (L:D) h. The larvae, used in the
feeding bioassays, had hatched after 4 Ð7 d; the larvae
were reared in this environmental chamber, during all
feeding trials.
Composition and Preparation of Diets. In this study,
eight artiÞcial diets, in four diet categories (diets
AÐH), and foliage (diet I) from the host plant L.
formosana were used to feed L. xylina larvae. The
compositions of all artiÞcial diets used in this study are
listed in Table 1. Diets A and B were modiÞed from the
gypsy moth high wheat germ diet (ODell et al. 1985),
with the primary component being wheat germ. The
main composition of diets C and D was wheat germ
and soybean powder, the formulations of these two
diets being modiÞed from the Spodoptera exigua
(Hübner) diet (L. C. Tang, unpublished). Diets E and
F were modiÞed from the S. litura diet (L. C. Tang,
unpublished), with the basic components being wheat
germ, soybean powder, and kidney bean powder. Fi-
nally, wheat germ and corn starch made up the prin-
cipal ingredients of diets G and H, which also had been
modiÞed from the S. litura diet (Kao 1995). In addi-
tion, to test the necessary of adding feeding stimulant,
L. formosana, leaf powder was added to the artiÞcial
diets B, D, F, and H. To prepare diets A and B, agar was
added into 150 ml of distilled water in a 1-liter beaker,
covered with foil, and autoclaved (121⬚C; ⬇15 min).
716 JOURNAL OF ECONOMIC ENTOMOLOGY
Cholesterol was then dissolved in ethyl ether and
blended with the main component wheat germ. On
the side, we mixed the other ingredients with 30 ml of
distilled water as the mingling solution. After auto-
claving, the agar was poured into the mixer and stirred
with the wheat germ (and leaf powder for diets B, D,
F, and H). The mingling solution was added into the
mixture as the agar was chilled to ⬇50⬚C. Then, we
distributed the still soft diet mixture into plastic rear-
ing cups (3 cm in diameter by 2.5 cm in height); each
cup contained food up to ⬇0.3 cm in height (to feed
the Þrst, second, and third instars). Alternatively, we
also poured the soften diet mixture into petri dishes
(14.5 cm in diameter by 1.5 cm in height to feed fourth
and older instars). The diet mixture was allowed to
solidify at room temperature and was kept in the
refrigerator until needed.
The preparation procedures for the other diet cat-
egories were similar to those of diets A and B. How-
ever, for diets C and D, cholesterol was blended with
wheat germ and soybean powder; for diets E and F,
cholesterol was blended with wheat germ, soybean
powder, and kidney bean powder; and cholesterol was
blended with wheat germ and corn starch for diets G
and H. The mingling solution combinations were also
slightly different.
Feeding Trials of L. xylina. To evaluate the quality
of different artiÞcial diets, as well as that of the host
plant, we conducted three types of insect bioassays: a
Þrst instar survival trial, and a long- and a short-term
feeding trial. First instar survival trials were conducted
to assess diet quality on the survivorship of L. xylina
larvae. In this bioassay, 10 newly hatched larvae were
placed into each plastic rearing cup, containing arti-
Þcial diet. Nine diets (eight artiÞcial diets [diet AÐH]
and one host plant [diet I]) were used in this trial, with
six replicates (10 larvae per replicate) for each diet.
After the larvae had been placed in each rearing cup,
containing one of the artiÞcial diets, it was covered
with a lid, inverted, and placed into the Percival cham-
ber (ODell and Rollinson 1966, ODell et al. 1985). For
the foliage treatment, we placed the larvae into petri
dishes (9 cm in diameter by 1.5 cm in height) con-
taining an excised L. formosana leaf. The leaf petiole
was wrapped with wet cotton, to maintain leaf turgor,
and leaves were changed every 1 to 2 d. After the
larvae had molted to the second instar, we recorded
the survival rates for each treatment.
The long-term feeding trial was conducted to assess
the effects of diets on insect development and growth
over the entire larval feeding, pupal, and adult stages.
In this bioassay, we only used the diets in which the
Þrst instars had survival rates of ⬎60% from the Þrst
instar survival trial (diets AÐF and I). One hundred
larvae were tested on each diet. Each assay consisted
of one newly hatched and weighed larva, placed into
a plastic rearing cup, with either artiÞcial diet or fo-
liage. To facilitate feeding, larvae were grouped (10)
in a cup and were individually fed when they reached
the third instar (ODell et al. 1985). The cup was
covered with a lid and then inverted in the Percival
chamber. Because of increased body size, we trans-
Vol. 99, no. 3
ferred fourth instars into petri dishes (5.5 cm in di-
ameter by 1.2 cm in height) and Þfth instars into even
larger petri dishes (9 cm in diameter by 1.5 cm in
height). To separate feces from diets, a suitably sized
piece of the artiÞcial diet was segmented and stuck
onto the inner side of the lid. The feeding methods for
diet I (foliage of L. formosana) were similar to the
procedures used in the Þrst instar survival trial. All
larvae were fed with corresponding diets until pupa-
tion and eclosion. We recorded both male and female
data on larval period, pupal period, pupal weight, and
adult weight. In addition, we also recorded larvae
survival rates and calculated the growth rate, based on
data from this trial. The growth rate formula, giving a
relative growth rate, and representing the mean
weight gain per day was growth rate ⫽ [ln(pupal
weight) ⫺ ln(hatching weight)] ⫼ (larval period)
(Nylin et al. 1993, Gotthard et al. 1994, Fischer and
Fiedler 2000).
The short-term feeding trial was conducted to eval-
uate the effects of diet quality on growth rates, food
consumption rates, and food processing efÞciencies of
fourth instars. In this bioassay, only the diets with
which larvae could successfully pupate, in the long-
term feeding trial, were used (diets A, E, and F, and
I). A group of larvae was reared on the gypsy moth
wheat germ diet (Cat. no. 960294, ICN Biomedicals
Inc., OH) from hatching until molting to the fourth
instar. Each assay consisted of a newly molted and
weighed fourth instar placed into a petri dish (5.5 cm
in diameter by 1.2 cm in height) containing a suitably
sized artiÞcial diet disk. For diet I (foliage of L. for-
mosana), the larvae were placed into the larger petri
dishes, similar to those used in the Þrst instar survival
trial, with the leaves being added to each dish. One
larva was used for each replicate, with 30 replicates for
each diet. We changed the food every 1 to 2 d, or as
necessary, during the bioassay. Upon molting to the
Þfth instar, the survival rate and fourth instar duration
were recorded; then, the larvae were frozen, oven-
dried at 40⬚C for 1 wk, and reweighed. Frass and
uneaten diet material also were collected, dried at
40⬚C for 1 wk, and weighed. Nutritional indices were
calculated to evaluate insect growth, consumption,
and food utilization efÞciency (Haynes and Millar
1998, Schoonhoven et al. 1998). These indices were
calculated from standard formulas for approximate
digestibility (AD), efÞciency of conversion of di-
gested food (ECD), efÞciency of conversion of in-
gested food (ECI), and total consumption (TC), as
described by Waldbauer (1968) and Haynes and Mil-
lar (1998). We used the initial, rather than the average
weights of larvae, to calculate relative growth rate
(RGR) and relative consumption rate (RCR) (Farrar
et al. 1989). Initial dry weights were estimated, based
on a wet-to-dry weight conversion factor, and deter-
mined from 10 newly molted fourth instars. Likewise,
the initial dry weight of the diets fed to larvae were
estimated by dry weight conversion, by using the suit-
able-sized piece, segmented from each artiÞcial diet
(diets A, E, and F) or the foliage collected from the
host plant (diet I), and dried at the time of the bio-
June 2006 SHEN ET AL.: L. xylina ON DIFFERENT DIETS
Fig. 1. Survival rate of Þrst instars of L. xylina reared with different diets (mean ⫾ SE) (TukeyÕs studentized range test).
AÐH, artiÞcial diets; I, host plant leaves of L. formosana. * Survival rate values are transformed to arcsine values when analyzed
by ANOVA.
assay. During the bioassay, a few larvae died because
of a virus infection. Because infected larvae were eas-
ily distinguished from the others, they were excluded
from our analyses.
Chemistry of Diets in Short-Term Feeding Trial. To
reveal the chemical components of the diets, we mea-
sured water and nitrogen content (nutrient manifes-
tations) from diets used in the short-term feeding trial
(diets A, E, and F, and I) because water and nitrogen
were the primary components provided for the cat-
erpillars (Zalucki et al. 2002). Concurrent with the
feeding trials, Þve pieces of each artiÞcial diet were
randomly segmented (in diet I, Þve pieces of foliage
of L. formosana were collected), weighed, oven-dried
at 40⬚C for 1 wk, reweighed, ground, and stored in a
freezer (⫺20⬚C). Water content and total nitrogen
content were quantiÞed for each diet sample. We
determined water content by weight difference be-
tween the wet and dry weights of the diet samples.
Diet nitrogen content was veriÞed by standard micro-
Kjeldahl assays. Diet samples were digested in acid
(Parkinson and Allen 1975), and nitrogen content was
quantiÞed by the micro-Nesslerization technique
(Lang 1958). Glycine p-toluene-sulfonic acid (5.665%
N) served as a nitrogen standard.
Statistical Analyses. In all trials, means and standard
errors of testing values were calculated by descriptive
statistics (PROC UNIVARIATE, SAS Institute 1999).
717
For the Þrst instar survival trial, survival rates were
analyzed by analysis of variance (ANOVA) (PROC
ANOVA, SAS Institute 1999), followed by compari-
sons of means using TukeyÕs studentized range (hon-
estly signiÞcant difference) test. We analyzed the test-
ing values in long- and short-term feeding trials, and
the chemistry of the diets, by ANOVA (PROC GLM,
SAS Institute 1999), followed by comparisons of
means by using least-signiÞcant difference (least sig-
niÞcant difference) test.
Results
First Instar Survival Trial. Results of this trial in-
dicated that the Þrst instar survival rate of L. xylina
varied signiÞcantly among insects reared with differ-
ent diets (Fig. 1). Except for diets G and H, Þrst instars
survived well (⬎60%) on most diets. Caterpillars fed
on diets E and F had the highest survival rate (100%);
no caterpillar survived beyond Þrst instar on diets G
and H, which were modiÞed from the S. litura diet
(Kao 1995).
Long-Term Feeding Trial. Because all Þrst instars
died when fed on diets G and H, these two diets were
excluded from the long-term feeding trial. All cater-
pillars, fed on diets B, C, and D died during the trial,
so these data were excluded from the analysis. The
result of the long-term feeding trial revealed that,
718 JOURNAL OF ECONOMIC ENTOMOLOGY Vol. 99, no. 3

overall, caterpillars grew better on their host plant

12.36 ⫾ 0.97d
15.89 ⫾ 0.97b
14.35 ⫾ 0.29c

17.69 ⫾ 0.15a
Growth rate
than on artiÞcial diets. The survival rate and growth

⬍0.001
(%/d)

3, 130
33.46
rate of L. xylina were signiÞcantly higher on the host
plant than on the artiÞcial diets (Table 2). Caterpillars
fed on the host plant (diet I) had the shortest larval
period and the highest growth rate (Table 2). The

760.87 ⫾ 106.95b
results also indicated that, except for the pupal period,

647.52 ⫾ 17.70b
1215.70 ⫾ 75.16a

566.90 ⫾ 0.00b
larvae performance varied signiÞcantly among the dif-
Female

⬍0.001
ferent diets (P ⬍ 0.001) (Table 2). Pupal and adult

41.02
3, 60
weights varied by 1.5- and 3-fold, respectively, among
Adult wt (mg)

the different diets (Table 2). In summary, larvae grew

well on the foliage of L. formosana (diet I), with the
highest survival and growth rates. Moreover, caterpil-
lars fed on diet A had the longest larval period and the
82.00 ⫾ 13.06b
239.52 ⫾ 21.79a

121.92 ⫾ 22.18a
199.26 ⫾ 6.54a

highest pupal and adult weights.

⬍0.001
19.98
Male

3, 58

Short-Term Feeding Trial. In this bioassay, we only

Performances of L. xylina reared with different diets in long-term feeding trails (mean ⴞ SE) (28°C:25°C; photoperiod of 12:12 关L:D兴)

tested those diets, with which the larvae could pupate

and eclose well, during the long-term feeding trial
(diets A, E, and F, and I). Results of the short-term
feeding trial revealed that performance (duration,
1,240.90 ⫾ 32.72b
1,213.11 ⫾ 27.08b
2,207.78 ⫾ 92.31a

1,141.60 ⫾ 0.00b

growth rate, and food processing efÞciency) of fourth

instars of L. xylina varied substantially among the
Female

⬍0.001
71.14
3, 62

different diets (Table 3). L. xylina larvae had RGR and

RCR values but a shorter duration on the host plant
Pupal wt (mg)

diet (diet I) than on artiÞcial diets (diets A, E, and F).

In contrast, the digestibility (AD) and conversion ef-
Þciencies (ECD and ECI) were higher for larvae fed
331.21 ⫾ 36.98d

613.10 ⫾ 15.05b
476.19 ⫾ 54.99c
961.53 ⫾ 81.71a

on artiÞcial diets. Of the three artiÞcial diets, A, E, and

⬍0.001

F, larvae fed on diet A had the best performance, with

Male

3, 67
32

the highest approximate digestibility (AD) and efÞ-

ciency of conversion of ingested food (ECI). Addi-
tionally, the survivorship of fourth instars of L. xylina
was ⬎75% in all diets tested. In general, the perfor-
mance of L. xylina, in this short-term study, paralleled
9.92 ⫾ 0.19

10.00 ⫾ 0.00
9.33 ⫾ 0.67
9.51 ⫾ 0.09
Female

Within a column, means bearing the same letter are not signiÞcantly different (P ⬎ 0.05).

that of insects in the long-term study, with faster

0.1986
3, 60
1.6

growth and higher consumption on host plant foliage.

Pupal period (d)

Diets and Host Plant Chemistry in Short-Term

Growth rate values are transformed to arcsine values when analyzed by ANOVA.

Feeding Trial. Water (P ⬍ 0.001) and nitrogen (P ⫽

0.0031) content of diets varied signiÞcantly (Table 4).
11.67 ⫾ 0.21

11.57 ⫾ 0.43
11.50 ⫾ 0.27
11.15 ⫾ 0.07

All artiÞcial diets had higher water (1.4-fold) and

0.0898
Male

3, 61
2.27

nitrogen (⬎2.4-fold) content than found in the host

plant. There was no signiÞcant difference in water and
nitrogen content among the artiÞcial diets.
46.00 ⫾ 0.00b
49.50 ⫾ 2.99b
47.80 ⫾ 0.41b
60.67 ⫾ 1.51a

Discussion
Female

⬍0.001
41.28
3, 62

In this study, it was clearly demonstrated that the

Larval period (d)

AÐF, artiÞcial diets; I, host plant L. formosana.

performance of L. xylina varied signiÞcantly among

different diets. Caterpillars fed on diets in which the
principal ingredient is corn starch cannot survive. In
48.50 ⫾ 3.94b

addition, our results also indicated that the necessity

40.22 ⫾ 0.55c
55.50 ⫾ 1.73a

59.56 ⫾ 4.11a

⬍0.001

of adding feeding stimulant (L. formosana powder) on

20.15
Male

3, 67

artiÞcial diets is still unclear.

The current literature indicates that some herbiv-
orous insects, such as Leucania separate Walker, Plute-
lla xylostella (L.), and Mamestra configurata Walker,
rate (%)
Survival

survived and grew better on host plants, than on ar-

29.51

8.14
14.06
87.76

tiÞcial diets (Hirai 1976, Hou and Hsiao 1986, Dosdall

0
0
0

and Ulmer 2004). The results of our study revealed a

Table 2.

similar pattern with survival rates and growth rates

increasing by 60 and 30%, respectively, in the host
Dieta

b
a

plant. Because this was the Þrst generation of the L.

Fb
df
D
C
A

E
B

P
I
June 2006 SHEN ET AL.: L. xylina ON DIFFERENT DIETS 719

Table 3. Performances of L. xylina reared with different diets in short-term feeding trails (mean ⴞ SE) (28°C:25°C; photoperiod of
12:12 关L:D兴)

RGR RCR
Dieta Survival rate (%) DUR (d) AD (%) ECD (%) ECI (%) TC (mg)
(mg/mg/d) (mg/mg/d)
A 95.83 7.75 ⫾ 0.21b 39.00 ⫾ 1.44a 59.69 ⫾ 4.11 22.18 ⫾ 0.96a 36.58 ⫾ 2.55c 0.16 ⫾ 0.01b 0.73 ⫾ 0.03b
E 76.92 9.70 ⫾ 0.59a 21.57 ⫾ 2.78bc 55.33 ⫾ 9.89 9.57 ⫾ 0.87b 56.24 ⫾ 5.33b 0.08 ⫾ 0.01c 0.85 ⫾ 0.08b
F 75.00 7.85 ⫾ 0.45b 24.02 ⫾ 1.85b 53.71 ⫾ 5.85 11.57 ⫾ 0.68b 52.68 ⫾ 3.75b 0.14 ⫾ 0.02b 1.14 ⫾ 0.08b
I 93.33 4.17 ⫾ 0.16c 16.60 ⫾ 1.93c 44.96 ⫾ 6.07 5.98 ⫾ 0.26c 101.95 ⫾ 9.71a 0.30 ⫾ 0.02a 5.06 ⫾ 0.35a
Fb 38.83 29.53 1.20 94.98 28.30 35.52 146.02
df 3, 61 3, 61 3, 61 3, 61 3, 61 3, 61 3, 61
P ⬍0.001 ⬍0.001 0.3180 ⬍0.001 ⬍0.001 ⬍0.001 ⬍0.001

AD, approximate digestibility; DUR, duration; ECD, efÞciency of conversion of digested food; ECI, efÞciency of conversion of ingested food;
RCR, relative consumption rate; RGR, relative growth rate; TC, total consumption.
Within a column, means bearing the same letter are not signiÞcantly different (P ⬎ 0.05).
a
A, E, and F, artiÞcial diets; I, host plant L. formosana.
b
AD, ECD, and ECI values are transformed to arcsine values when analyzed by ANOVA.

xylina to feed on artiÞcial diets, the insects may need
more time to adapt to the artiÞcial diets (Kao 1995).
According to Schoonhoven et al. (1998), plants are
suboptimal food, because of the diluted nutrients in a
medium of indigestible compounds. In response to
low-quality foods, herbivorous insects may have com-
pensatory reactions, such as increased food consump-
tion and nutrient utilization efÞciency (Timmins et al.
1988, Wheeler and Halpern 1999, Awmack and
Leather 2002, Chen et al. 2004), that result in a Þnal
biomass equivalent to larvae fed on high-quality food
(Slansky and Feeny 1977). The foliar nitrogen content
of L. formosana was lowest for all diets tested, and the
caterpillars consumed signiÞcantly more L. formosana
biomass, than that consumed in the other diets. Thus,
our results suggest that compensatory feeding may
occur in L. xylina, in response to poor-quality host
plant foliage.
Comparisons of the performance of L. xylina,
among the various artiÞcial diets, revealed differences
in their ability to effectively use the diets. Because
different insect species often vary to some extent in
their speciÞc nutritional requirements, small changes
in diet composition may have drastic effects on insect
performance (Schoonhoven et al. 1998). The larvae of
L. xylina did not feed well on diet G, which was
modiÞed from the S. litura diet (Kao 1995). In contrast,
Table 4. Water and nitrogen content in different diets (mean ⴞ
SE)
Dieta Water content (%) Nitrogen content (%)
A 81.86 ⫾ 0.04a 5.00 ⫾ 1.24a
E 81.94 ⫾ 0.03a 4.28 ⫾ 0.92a
F 82.23 ⫾ 0.06a 3.05 ⫾ 0.31a
I 56.14 ⫾ 1.11b 1.26 ⫾ 0.18b
Fb 646.05 7.05
df 3, 19 3, 19
P ⬍0.001 0.0031
Within a column, means bearing the same letter are not signiÞcantly
different (P ⬎ 0.05).
a
A, E, and F, artiÞcial diets; I, host plant L. formosana.
b
Water and nitrogen content values are transformed to arcsine
values when analyzed by ANOVA.
caterpillars grew very well on another diet, which had
also been modiÞed from the S. litura diet (diet E)
(L. C. Tang, unpublished). The composition of these
two diets was very similar and the cause of the sig-
niÞcant difference is unknown. Our results also re-
vealed that caterpillars fed on diet C, which was mod-
iÞed from the S. exigua diet (L. C. Tang, unpublished)
could not successfully pupate. The reason for this is
also unclear, but diet composition may play an impor-
tant role. The main protein source in diet C was wheat
germ; past studies have revealed that the protein
source is a critical factor inßuencing insect growth
(Schoonhoven et al. 1998, Cohen 2004). It also has
been found that S. exigua and Helicoverpa zea (Bod-
die) reared on a synthetic diet containing animal pro-
tein (casein) had superior growth compared with
those fed on diets containing vegetable protein (soy-
bean powder) (Duffey et al. 1986). Our results
showed that the caterpillars grew best on diet A, which
also contained casein as the main protein source ex-
cept from wheat germ.
Concerning the necessity of adding feeding stimu-
lant, we added leaf powder of L. formosana on several
diets. The results revealed that where adding leaf
powder makes the diet better is still questionable.
Caterpillars of L. xylina performed no worse or some-
what better on diets with leaf powder added. Larvae
used in this study were from Þeld-collected eggs, and
they had no experience on artiÞcial diets. Therefore,
these caterpillars may have difÞculty to adapt the
artiÞcial diets. Our long-term feeding trial indicated
that the overall survival rates were signiÞcantly low on
all the artiÞcial diets. This low survival may mask the
effects of leaf powder.
In addition, the water and nitrogen content in the
diets of herbivores have been considered as important
factors affecting their performance (Schoonhoven et
al. 1998). Schoonhoven et al. (1998) revealed that in
caterpillars, the developmental period was longer
when higher nutrient food was provided; the cater-
pillars seemed need more time to digest and absorb the
nutrient, and the Þnal pupal weight was heavier than
those fed on poor nutrient diet. Timmins et al. (1988)
720 JOURNAL OF ECONOMIC ENTOMOLOGY
also indicated that the feeding time for caterpillars was
longer on diets having an elevated water content than
on normal water content diets. Our results were sim-
ilar to those of others in that diet (diet A), which
contained higher water and nitrogen, was the more
nutritious diet. Moreover, concerning the unit price of
the artiÞcial diets, we found that the unit price of diet
A (1.78 NT$/g dry material) was less than that of diet
E (2.04 NT$/g dry material) or F (2.03 NT$/g dry
material). Combining this information, with the re-
sults of the feeding performances, it was determined
that diet A could be the best diet of those we tested
for artiÞcial rearing of L. xylina caterpillars.
In summary, this research has indicated that diet
composition has a signiÞcant effect on L. xylina per-
formance and that the diet, modiÞed from the gypsy
moth diet, was the most suitable for L. xylina. In
addition, larvae reared on the host plant grew faster
than those reared on artiÞcial diets. We also found that
size of laboratory-reared adults was signiÞcantly larger
than Þeld-captured adults and these laboratory-reared
adults can successfully reproduce in laboratory (S.-
Y.H., unpublished data). However, this study is only a
Þrst step. For future mass rearing purposes, the diet
composition and ratios may require Þne-tuning to im-
prove insect survive and performance.
Acknowledgments
We thank L. C. Tang for valuable suggestions in preparing
artiÞcial diets and rearing methods and R. Haesevoets for
comments on the manuscript. Two anonymous reviewers
provided helpful comments. This research was supported by
Grant 92AS-1.7.2-BQ-B2 from the Bureau of Animal and
Plant Health Inspection and Quarantine, Council of Agri-
culture, Taiwan (to S.-Y.H.).
References Cited
Awmack, C. S., and S. R. Leather. 2002. Host plant quality
and fecundity in herbivorous insects. Annu. Rev. Ento-
mol. 47: 817Ð844.
Bailey, C. G. 1976. A quantitative study of consumption and
utilization of diets in the bertha armyworm, Mamestra
configurata (Lepidoptera, Noctuidae). Can. Entomol.
108: 1319Ð1326.
Chang, Y. C., and Y. C. Weng. 1985. Morphology, life habit,
outbreak and control of casuarina tussock moth (Lyman-
tria xylina Swinhoe). Q. J. Chin. For. 18: 29Ð36.
Chao, J. T., P.W.Schaefer, Y. B. Fan, and S. S. Lu. 1996. Host
plants and infestation of casuarina moth Lymantria xylina
in Taiwan. Taiwan J. For. Sci. 11: 23Ð28.
Chen, Y. Z., L. Lin, C. W. Wang, C. C. Yeh, and S. Y. Hwang.
2004. Response of two Pieris (Lepidoptera: Pieridae)
species to fertilization of a host plant. Zool. Stud. 43:
778Ð786.
Cohen, A. C. 2004. Insect diets: science and technology.
CRC, New York.
Dosdall, L. M., and B. J. Ulmer. 2004. Feeding, develop-
ment, and oviposition of bertha armyworm (Lepidoptera:
Noctuidae) on different host plant species. Environ. En-
tomol. 33: 756Ð764.
Duffey, S. S., K. A. Bloem, and B. C. Campbell. 1986. Con-
sequences of sequestration of plant natural products in
Vol. 99, no. 3
plant-insect-parasitoid interactions, pp. 31Ð 60. In D. J.
Boethel and R. D. Eikenbary [eds.], Interactions of plant
resistance and parasitoids and predators of insects. Ellis
Horwood, Chichester, United Kingdom.
Farrar, R. R., Jr., J. D. Barbour, and G. G. Kennedy. 1989.
Quantifying food consumption and growth in insects.
Ann. Entomol. Soc. Am. 82: 593Ð598.
Fischer, K., and K. Fiedler. 2000. Response of the copper
butterßy Lycaena tityrus to increase leaf nitrogen in nat-
ural food plant: evidence against the nitrogen limitation
hypothesis. Oecologia (Berl.) 124: 235Ð241.
Gotthard, K., S. Nylin, and C. Wiklund. 1994. Adaptive vari-
ation in growth rate: life history costs and consequences
in the speckled wood butterßy, Pararge aegeria. Oecolo-
gia (Berl.) 99: 281Ð289.
Gray, D. R., F. W. Ravlin, and J. A. Braine. 2001. Diapause
in the gypsy moth: a model of inhibition and develop-
ment. J. Insect Physiol. 47: 173Ð184.
Gries, G., P. W. Schaefer, G. Khaskin, R. Hahn, R. Gries, and
J. T. Chao. 1999. Sex pheromone components of casu-
arina moth, Lymantria xylina. J. Chem. Ecol. 25: 2535Ð
2545.
Haynes, K. F., and J. G. Millar. 1998. Methods in chemical
ecology, vol. 2. Chapman & Hall, New York.
Hirai, K. 1976. A simple artiÞcial diet for mass rearing of the
armyworm, Leucania separata Walker (Lepidoptera:
Noctuidae). Appl. Entomol. Zool. 11: 278 Ð283.
Hou, R. F., and M. L. Hsiao. 1986. An improved diet for
rearing the diamondback moth, Plutella xylostella and its
requirements for fatty acids. Chin. J. Entomol. 6: 31Ð37.
Hwang, S. Y., C. C. Jeng, T. C. Shen, Y. S. Shae, and C. S. Liu.
2004. Diapause termination in casuarinas moth (Lyman-
tria xylina) eggs. Formos. Entomol. 24: 43Ð52.
Kao, S. S. 1995. Insect mass rearing, pp. 1Ð 8. In K. C. Lee
[ed.], Taiwan Agricultural Chemicals and Toxic Sub-
stances Research Institute Special Report 37. Taichung,
Taiwan.
Lang, C. A. 1958. Simple microdetermination of Kjedahl
nitrogen in biological materials. Anal. Chem. 30: 1692Ð
1694.
Nylin, S., C. Wiklund, P. O. Wickman, and E. Garcia-Barros.
1993. Absence of trade-offs between sexual size dimor-
phism and early male emergence in a butterßy. Ecology
74: 1414 Ð1427.
ODell, T. M., and W. D. Rollinson. 1966. A technique for
rearing the gypsy moth, Porthetria dispar (L.), on an
artiÞcial diet. J. Econ. Entomol. 59: 741Ð742.
ODell, T. M., C. A. Butt, and A.W. Bridgeforth. 1985. Ly-
mantria dispar, pp. 355Ð367. In P. Singh and R. F. Moore
[eds.], Handbook of insect rearing, vol. II. Elsevier, New
York.
Parkinson, J. A., and S. E. Allen. 1975. A wet oxidation pro-
cedure suitable for the determination of nitrogen and
mineral nutrients in biological material. Commun. Soil
Sci. Plant Anal. 6: 1Ð11.
SAS Institute. 1999. UserÕs manual, version 8.0. SAS Insti-
tute, Cary, NC.
Schoonhoven, L. M., T. Jermy, and J.J.A. van Loon. 1998.
Insect-plant biology from physiology to evolution. Chap-
man & Hall, New York.
Shen, T. C., Y. S. Shae, C. S. Liu, C. W. Tan, and S. Y. Hwang.
2003. Relationships between egg mass size and egg num-
ber per egg mass in the casuarina moth, Lymantria xylina
(Lepidoptera: Lymantriidae). Environ. Entomol. 32: 752Ð
755.
Slansky, F., and P. Feeny. 1977. Stabilization of the rate of
nitrogen accumulation by larvae of the cabbage butterßy
June 2006 SHEN ET AL.: L. xylina ON DIFFERENT DIETS
on wild and cultivated food plants. Ecol. Monogr. 47:
209 Ð228.
Timmins, W. A., K. Bellward, A. J. Stamp, and S. E. Reynolds.
1988. Food intake, conversion efÞciency, and feeding
behaviour of tobacco hornworm caterpillars given arti-
Þcial diet of varying nutrient and water content. Physiol.
Entomol. 13: 303Ð314.
Waldbauer, G. P. 1968. The consumption and utilization of
food by insect. Adv. Insect Physiol. 5: 229 Ð288.
721
Wheeler, G.S.S., and M. D. Halpern. 1999. Compensatory
responses of Samea multiplicalis larvae when fed leaves of
different fertilization levels of the aquatic weed Pistia
stratiotes. Entomol. Exp. Appl. 92: 205Ð216.
Zalucki, M. P., A. R. Clarke, and S. B. Malcolm. 2002. Ecol-
ogy and behavior of Þrst instar larval Lepidoptera. Annu.
Rev. Entomol. 47: 361Ð393.
Received 1 July 2005; accepted 20 February 2006.