J. stored Prod. Res., 1973, Vol. 8, pp. 291-297. Pergamon Press. Printed in Great Britain.

Effect of Moisture on Growth Rate

and Development of Two Strains of
Tenebrio molitor L. (Coleoptera,
Tenebrionidae)
K. C. D. URS* and T. L. HOPKINS
Department of Entomology, Kansas State University, Manhattan, Kansas 66506, U.S.4.

(First received 18 Jub 1972, and infinalform 19 September 1972)

Abstract-Larvae of Tenebrio molitor L., reared on dry wheat shorts, grew and developed
much more rapidly when free-choice water was available than when it was not. When
water was supplied in cotton pads, larvae often burrowed in the moist pads and could
obtain water by cuticular absorption or ingestion. Heavier larvae were also produced at
pupation with a tendency for reduction of larval and pupal mortality and an increase
in adult longevity in the water regimen. Two strains of T. molitol- showed highly sig-
nificant differences in their rates of growth and development. The faster developing
strain in the dry condition also developed faster when free-choice water was available
and weighed more as prepupal larvae, pupae and adults. Water also reduced the number
of molts in the faster-developing strain. Although the mechanisms enabling more rapid
growth under both dry and moist conditions are not known, strain differences in
mealworms indicate genetic potential for completing life cycles more rapidly in dry
environments with extremely rapid development possible under optimum moisture
conditions.
INTRODUCTION

ENVIRONMENTALmoisture, whether as a liquid which can be ingested or absorbed

through the cuticle, or as atmospheric water vapor, is critical for stored products
insects, which typically inhabit water-deficient environments. Several species of
stored products insects have been shown to grow and develop more rapidly at high
than at low humidities (ANDREWARTHA and BIRCH, 1954; BURSELL,1964). Although
yellow mealworm larvae resist desiccation and can survive long periods at 0 per cent
r.h. (SCHULZ, 1930; BUXTON, 1932), they seemingly prefer moist situations (COTTON
and ST.GEORGE, 1929). Perhaps larval preference for moisture is related to their
ability to absorb water through the cuticle at r.h. of 90 per cent or more, while water
losses occur at lower relative humidities (BUXTON, 1930; MELLANBY, 1932). Larvae
also will drink water when available and grow and develop faster as a consequence
(MELLANBY and FRENCH, 1958; MURRAY, 1968).
We have determined the effects of abundant water in addition to that contained
in dry food on growth, development, life cycle completion and adult longevity of
* Present address : Department of Entomology, University of Agricultural Sciences, Hebbal, Banga-
lore-24, India.
291
 292 K. C. D. URS and T. L. HOPKINS

two strains of yellow mealworm, Tenebrio molitor L. Throughout the life cycle, water
was supplied continuously (on a free-choice basis) in cotton pads separate from the
food. Moist pads offered an opportunity for larvae both to drink and to absorb
water through the cuticle from a microclimate of high r.h.

MATERIALS AND METHODS

Insect cultures
Cultures of Tenebrio molitor L. (adults and larvae) were obtained originally from
two sources. Strain A came from the U.S. Department of Agriculture Midwest
Grain Investigations Laboratory, Manhattan, Kansas; Strain B originated from
The Chemagro Corporation, Kansas City, Missouri. Stock cultures of each strain
were maintained on feeding regimens of dry food (screened wheat shorts) and dry
food plus water (distilled water provided on cotton pads). Fresh wheat shorts were
provided once a month, and the cotton pads were changed every three days to prevent
mold growths in the cultures. Dead mealworms, if any, were removed.

Growth, development and l;fe cycles

Growth and development studies were conducted by rearing both strains on
screened wheat shorts with or without water in plastic dishes (3 -8 cm high and
150 cm dia) with plastic mesh tops. Water was provided daily in a pad of wet cotton
placed away from the food to prevent mold development. Fresh food was given
every 3 weeks. Stock and experimental cultures were held in a room maintained at
80°F (26.7%) 50 per cent r.h., with a photoperiod of 16 hr light and 8 hr dark.
To obtain insects the same age for each experiment, newly hatched larvae were
collected over an interval of a few hours. Adult beetles of both sexes were confined
in plastic dishes with wheat flour and pieces of fresh lettuce, carrot, or potato pro-
vided on black paper to facilitate observation and egg collection. Adults were
removed after about a week and eggs were retained in the respective dishes for
hatching. For each replicate experiment, batches of 30-50 larvae hatched the
same day, were used. Once a week an entire group of larvae was weighed and the
average weight calculated. Growth curves were plotted from averages of three repli-
cate experiments. The larval period was calculated as the average number of days
to pupation for the individual insects. Body lengths of prepupal larvae (identified by
their inactivity and curved posture) were determined by measuring individuals
under CO, anesthesia.
The number of larval instars was determined by rearing ten newly hatched larvae
of each strain individually under each regimen in small plastic dishes. Three to
four grams of finely sieved wheat flour were given twice in the first 2 weeks; there-
after screened wheat shorts were given once every 3 weeks until pupation. For the
water regimen, cotton pads dipped in distilled water were provided daily. Daily
until pupation, larval molts for each individual were observed and recorded.
Individual weights of pupae were recorded directly after pupation. The pupal period
for each individual was determined by recording the number of days from pupation
to ecdysis. Individual weights of adults were recorded directly after ecdysis. Length
of life cycle was determined by the total days from egg to pupal-adult ecdysis.
Data were analyzed for significance between strains and treatments (water and
no water) by two-way analysis of variance. When significant interaction was en-
countered, the differences were analyzed by LSD.
 Tenebrio molitor L. 293

RESULTS

Larval growth and development

Larval growth rates of both strains were similar through the first 20 days (Fig. 1).
Subsequently both strains supplied with water showed accelerated growth rates over
those without water.

AGE - DAYS

FIG. 1. Larval growth curves of two strains (A and B) of the yellow mealworm Tenebrio
molitor L., reared with or without abundant water. Points on curves represent averages
of replicate experiments of 30-50 larvae each. After the start of pupation, points represent
decreasing numbers of larvae.

Strain B grew more rapidly than strain A on both dry food and water regimens.
Strain B larvae supplied with water had an average larval period of 88 days with
pupation starting at 64 days. In contrast, strain A larvae had an average larval
period of nearly double that of strain B with pupation starting in 105 days. In the
dry food regimen, strain B larvae also showed faster growth rates than strain A larvae,
with a 155 day larval period and pupation beginning at 135 days. Strain A larvae
began pupating at 153 days and spent an average of 202 days as larvae. Therefore,
water accelerated growth and development of Tenebrio molitor confirming the results
of MELLANBY and FRENCH (1958). In addition, strain B larvae grew and developed
faster than strain A larvae in both regimens. Differences between strains and treat-
ments as to average larval period and time of first pupation were highly significant
(Table 1).
Mealworms of strain A averaged 16 larval instars in both dry food and the water
regimens (Table 1). In strain B, there were 15 instars in dry food and 12 in the water
regimen. Water did not influence the number of instars in strain A but was highly
significant in reducing number of instars as well as larval period in strain B. There-
fore, depending on the strain, moisture may or may not reduce number of instars
as it speeds growth and development. COTTON and ST. GEORGE (1929) reported a
range of 9-20 larval instars for Tenebrio molitor L. ; of the 35 specimens tested, half
molted 17-19 times each.
 TABLE 1. LIFE CYCLESOF TWO STRAINS(A AND B) OF Tenebrio molitor L., REARED U?TH OR WITHOUT WATER

Average number of days and instars’

Strain A Strain B Statistical significancea
Stages No water Water No water Water Strain Treatment P
h
-__
Larval period 202.5 159.0 155.4 87.7 ** *x &
First pupation 153.5 105.2 134.7 64.0 ** **
Pupal period 10.1 8.5 9.6 7.7 ns * ti
Total preimaginal period 213.3 177.3 166.1 99.2 ** ** B
Range 170.8-267.0 134.0-242.3 149.3-188.0 79.0-125.3
;
Adult longevity 25.0 60.73 31.3 105.23 t t
Larval instars r
(number and range) 16.0 16.1 14.9 12.23
(15-17) (13-18) (14-16) (11-14)

1 Averages of three replicates. Adult longevity and number of larval instars, one test of 30 adults and 10 larvae each strain and treatment.
a * P < 0.05; ** P < 0.01; ns nonsignificant; i interaction significant, ** ; two-way analysis of variance.
8 Significantly different from other groups in the experiment, P = 0.05.
 Effect of Moisture on Growth Rate of TenebriomolitorL. 295

Prepupal larval weights of both strains were significantly higher in the water
regimen than in dry food alone (Table 2). Strain B prepupae were significantly
heavier than strain A prepupae when water was supplied but no difference was
observed in the dry food regimen. Average body lengths of prepupal larvae were
also greater when water was available. Strain B were longer than strain A under
both regimens (Table 2). Percentage of larval mortality (Table 3) was significantly
higher for strain A than for strain B under both dry and wet conditions. Availability
of water tended to reduce larval mortality although this was not significant due to
replicate variability.
T.XBLE 2. LIVE WEIGHT AND BODY LENGTH OF TWO STRAINS (A AND B) OF Tenebriomlitor L. REARED
WITHOR WITHOUT WATER

Weight in mg’ and body length in cm2

Strain A Strain B Statistical significance3
Stages No water Water No water Water Strain Treatment

Prepupal larvae 100.6 113.6* 101.7 129.74 t t

Pupae 94.5 114.3 108.4 128.3 ** **
Adults 80.0 97.2 91.5 107.1 ** **
Prepupal body length 2.1 2.4 2.2 2.5 ** **

r Averages of three replicates.

3 One test only of 25 prepupal larvae per strain and treatment.
3 * P < 0.05; P < 0.01; t interaction significant, * ; two-way analysis of variance.
4 Significantly different from other groups in the experiment, P = 0.05.

TABLE 3. PERCENTAGEOFMORTALITYANDADULTEMERGENCEOFTWOSTRAINS (A AND B) OF Tenebrio

??ZO&OrL. DURINGDEVELOPMENTWITHORWITHOUTWATER

Percentage
Strain A Strain B Statistical significancea
Stages No water Water No water Water Strain Treatment

Larval mortality 30.6 26.5 19.1 Il.0 * ns

Pupal mortality 25.4 14.4 10.5 8.6 ns ns
.4dult emergence 51.6 63.4 72.7 81.5 * ns

r Average of three replicates.

2 * P < 0.05; ns nonsignificant. Two-way analysis of variance.

Pupal period
The average pupal periods were significantly shorter for both strains when water
was available but no significant difference was found between strains. COTTON and
ST. GEORGE (1929) recorded a pupal period ranging from 7 days (at 81 OF) to 18
days (at 65°F) for mealworms.
Pupae of both mealworm strains were heavier when larvae were grown with water
than without, corresponding to the larval weights from the two rearing conditions.
Average pupal weights of strain B were significantly greater than those of strain A
under both wet and dry regimens (Table 2). No significant differences in pupal
mortality due to strain or treatment differences were observed (Table 3).
 K. C. D. URS and T. L. HOPKINS

Adult emergence and longevity

The availability of water had a highly significant effect on reducing the total
length of the pre-imaginal period in both strains of mealworms (Table 1).
The percentage of adults emerging was significantly higher in mealworms of
strain B than in strain A, both with and without access to water (Table 3). However,
apparent increases in adult emergence when water was available were nonsignificant.
Average adult weights were much higher when the two mealworm strains were
given water than dry food alone (Table 2). In both rearing conditions adults of
strain B were heavier than adults of strain A. In general, there was a weight loss
between pupation and adult emergence.
Adults of both strains survived longer when supplied with water (Table 1). Adult
longevity was markedly higher for strain B than for strain A in the water regimen.

DISCUSSION
Mealworm larvae are very resistant to desiccation and can exist for long periods
in extremely dry situations (SCHULZ, 1930; BUXTON, 1932). During exposure to
desiccation, growth may cease, and if larvae are not ready to pupate, a long period
of dormancy follows (SCHULZ, 1930) in which the larvae may cease feeding entirely
(MURRAY, 1968). Under natural conditions, however, mealworm larvae are found
primarily in damp situations which they seem to prefer (COTTON and ST. GEORGE,
1929).
In our studies, mealworm larvae from two strains grew and developed much more
rapidly when water was available on a free-choice basis than when it was not. Larvae
reared with water also weighed more just before pupation; and this weight differential
was carried over to the pupal and adult stages. The increase in weight was partially
attributed to an increase in stored lipids, there being little difference in total body
water (URS and HOPKINS, 1973). Thus, by actively seeking moist environments
larvae greatly speed maturation while still retaining the mechanisms to resist desic-
cation if moisture becomes scarce. In moist situations water intake can occur through
the cuticle as well as by ingestion. BUXTON (1930) and MELLANBY (1932) have
demonstrated gain of water by cuticular absorption in r.h. of 90 per cent or more.
Such uptake of water vapor may occur mainly through the rectal cuticle rather than
through the general body surface (NOBLE-NESBITT, 1970). We have observed that
mealworm larvae commonly burrow and rest in moist cotton pads, indicating they
prefer that environment when not feeding in the dry food. Water uptake could
readily occur through the cuticle in such microclimates of high r.h. Ingesting water
is another possibility for uptake especially when wet pads are first placed in the
rearing chambers. MELLANBY and FRENCH (1958) and MURRAY (1968) have shown
that periodic drinking of water by mealworms speeded growth and development.
We observed considerable differences in the abilities of two strains of yellow meal-
worms to grow and develop under both dry and moist conditions. Though the faster
developing strain (B) in both regimens apparently had more efficient mechanisms
for utilizing nutrients to grow and develop, the exact nature of the phenomenon is
unknown. LECLERCQ (1963) by selecting T. molitor pupae for heavy and light weights
obtained strains with shorter and longer larval periods and other physiological
differences. Our strain B synthesized more lipids per unit weight during larval life
than did the slower developing strain A (URS and HOPKINS, 1973).
 Effect of Moisture on Growth Rate of Tenebrio molitor L. 297

Little is known about how desiccation slows larval growth in insects or how
moisture promotes it. ScHuLz (1930) mentioned that much more undigested food
was in the feces of mealworms living under dry conditions, indicating that digestion
and absorption of nutrients could be impaired by reducing salivary secretions and
moisture in the gut contents. Consequently, energy production, uptake of essential
nutrients for growth and storage of lipids, as we have shown, would be curtailed.
FRAENKEL and BLEWETT (1944) concluded that several stored product insects reared
at low r.h. must use more food for metabolic water production and less for increases
in dry matter. LECLERCQ (1948) obtained similar results with T. molitor. MURRAY
(1968) has shown that under very dry conditions Tenebrio larvae may cease feeding
and become inactive until moisture is again available. In the dry regimen we in-
vestigated, sufficient moisture was available in the food to promote continuous
growth and development. Strain differences indicate that mealworms have genetic
potential to complete life cycles more rapidly under dry conditions with extremely
rapid development under optimum moisture conditions.

Acknowledgements-We are indebted to Dr. ARTHUR DAYTON and Mr. L. J. KRCHMA for assistance with
statistical analysis of data and to Mr. KRCHMA for critical reading of the manuscript.
This work is Contribution No. 1082, Department of Entomology, Kansas Agricultural Experiment
Station, Manhattan, Kansas. The research was supported in part by U.S. Department of Agriculture,
Market Quality Research Division, Stored Products Insects Research Branch, Grant 12-14-IOO-
9981(51) to T.L.H.
The Predoctoral studies of K.C.D.U. were supported by a Ford Foundation Fellowship.

REFERENCES
ANDREM~ARTHA,H. G. and BIRCH, L. C. (1954) The distribution and abundance of animals. Univ. Chicago
Press.
BURSELL,E. (1964) Environmentalaspects: humidity in, ThePhysiologyofZnsecta. (Ed. by RCCKSTEIN, M.)
1, 323-361. Academic Press, New York.
BUXTON, P. A. (1930) Evaporation from the mealworm and atmospheric humidity. Proc. R. SOL. (1%)
106, 560-577.
BUXTON, P. A. (1932) Terrestrial insects and the humidity of the environment. BioZ. Rev. 7, 275-320.
COTTON, R. T. and ST. GEORGE, R. A. (1929) The mealworms. Tech. Bull. U.S. Dep. Agric. 95, I-38.
FRAENICEL,G. and BLEWETT, M. (1944) The utilization of metabolic water in insects. Bull. ent. Res.
35, 127-139.
LECLERQ, J. (1948) Contribution a l’&ude du metabolisme de I’eau chez la larve de Tenebrio molitor
L. Archs int. Physiol. 55, 412-419.
LECLERQ, J. (1963) Artificial selection for weight and its consequences in Tenebrio molitor L. Nature,
Lond. 198, 106-107.
MELLANRY, K. (1932) The effect of atmospheric humidity on the metabolism of the fasting meal-
worm (Tenebrio molitor L. Coleoptera). Proc. R. Sot. (B) 111, 376-390.
MELLANHY, K. and FRENCH, R. A. (1958) The importance of drinking water to larval insects.
Entomologie exp. appL. 1, 116-124.
MURRAY, D. R. P. (1968) The importance of water in the normal growth of larvae of Tenebrio molitor
L. Entomologia exp. appl. 11, 149-168.
NOBLE-NESBITT, J. (1970) Water balance in the firebrat, Thermobia domestica (Packard). The site of
uptake of water from the atmosphere. J. ext. Biol. 52, 193-200.
SCHULZ, F. N. (1930) Zur Biologie des Mehlwurms ( Tenebrio molitor). I. Der Wasserhaushalt. Biochem.
2. 227, 340-353.
URS, K. C. D. and HOPKINS, T. L. (1973) Effect of moisture on the lipid content and composition of
two strains of Tenebrio molitor L. (Coleoptera, Tenebrionidae). J. stored Prod. Res. 8,299-305.