Trichogramma cacoeciae Marchall, T. brassicae Bezdenko and T. evanescens Westwood (Hymenoptera: Trichogrammatidae) could be useful in biological control programs of agricultural insect pests. The possibility of storing Trichogramma species at low temperatures and the effect of such storage on the quality of the parasitoids and their fecundity were studied. Trichogramma cacoeciae, T. brassicae and T. evanescens pupae were stored 1, 2, 3, and 4 weeks at 4 ± 1 °C in a refrigerator, 60-70% R.H. and full darkness. Parasitoid emergence was 98.80%, 99.33% and 99.60% for T. cacoeciae, T. brassicae and T. evanescens, respectively, after 1 week of storage. Storage at 4 ± 1 °C resulted in a significant decline in parasitoid emergence after 3 weeks. Subsequent trials focused on fitness of stored pupae in terms of percentage of parasitized eggs and longevity of females. Storage at 4 ± 1°C reduced fecundity and longevity of female parasitoids

Ozder, N.

Saglam, O.

English

ValpoScholar

The Great Lakes Entomologist 
Volume 37 
Numbers 3 & 4 - Fall/Winter 2004 Numbers 3 & 
4 - Fall/Winter 2004 
Article 9 
October 2004 
Effect of Short Term Cold Storage on the Quality of Trichogramma 
Brassicae, T. Cacoeciae, and T. Evanescens (Hymenoptera: 
Trichogrammatidae) 
N. Ozder 
Trakya University 
O. Saglam 
Trakya University 
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Ozder, N. and Saglam, O. 2004. "Effect of Short Term Cold Storage on the Quality of Trichogramma 
Brassicae, T. Cacoeciae, and T. Evanescens (Hymenoptera: Trichogrammatidae)," The Great Lakes 
Entomologist, vol 37 (2) 
Available at: https://scholar.valpo.edu/tgle/vol37/iss2/9 
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2004 THE GREAT LAKES ENTOMOLOGIST 183
EFFECT OF SHORT TERM COLD STORAGE ON THE QUALITY OF
TRICHOGRAMMA BRASSICAE, T. CACOECIAE, AND T. EVANESCENS
(HYMENOPTERA: TRICHOGRAMMATIDAE)
N. Özder1  and Ö. Saglam
ABSTRACT
Trichogramma cacoeciae Marchall, T. brassicae Bezdenko and T. evanescens
Westwood (Hymenoptera: Trichogrammatidae) could be useful in biological con-
trol programs of agricultural insect pests.  The possibility of storing Trichogramma
species at low temperatures and the effect of such storage on the quality of the
parasitoids and their fecundity were studied.  Trichogramma cacoeciae, T. brassicae
and T. evanescens pupae were stored 1, 2, 3, and 4 weeks at 4 ± 1 °C in a refrigera-
tor, 60-70% R.H. and full darkness.  Parasitoid emergence was 98.80%, 99.33%
and 99.60% for T. cacoeciae, T. brassicae and T. evanescens, respectively, after 1
week of storage.  Storage at 4 ± 1 °C resulted in a significant decline in parasitoid
emergence after 3 weeks.  Subsequent trials focused on fitness of stored pupae in
terms of percentage of parasitized eggs and longevity of females.  Storage at 4 ±
1°C reduced fecundity and longevity of female parasitoids.
____________________
Egg parasitoids of the genus Trichogramma (Hymenoptera:
Trichogrammatidae) are among the most commonly used natural enemies in the
world against lepidopteran pests (Hassan 1993, King 1993, Smith 1996).  They
are gregarious endoparasitoids with the majority of species attacking eggs of
Lepidoptera, although some also have the potential to attack dipteran and co-
leopteran eggs (Hoffmann et al. 1955, Mansfield and Mills 2002).  Codling moth
(Cydia pomonella, L.) (Lep., Tortricidae), grapevine moth (Lobesia botrana, Den.
and Schiff.) (Tortricidae) and European leafrollers (Archips rosanus, L.) (Tortricidae)
have been considered potential targets for Trichogramma inundation in orchard
crops (Hassan 1992, Cossentine and Jensen 2000, Hommay et al. 2002).
Trichogrammatid egg parasitoids are generally used for inundative bio-
logical control, as annual releases.  Storage of natural enemies assures their
availability in sufficient number at the time of release.  Storage techniques
must ensure the availability of high quality natural enemies (Bigler 1994).
Therefore, the development of storage techniques for biocontrol agents is consid-
ered of utmost importance to provide flexibility and efficiency in mass produc-
tion, to synchronize a desired stage of development for peak release and to make
available standardized stocks for use in research (Greco and Stilinovic 1998,
Leopold 1998).  Although there are numerous investigations about storage of
trichogrammatid species (Jalali and Singh 1992, Greco and Stilinovic 1998,
Pitcher et al. 2002, Tezze and Botto 2004), natural variation among species
necessitates individual studies to determine species-specific parameters.  The
effect of different periods of cold storage on the quality of the stored parasitoids
was evaluated by measuring the following variables: adult emergence time,
adult emergence proportion, female proportion, fecundity and longevity.
Thus, the aim of this research was to determine whether T. cacoeciae
Marchall, T. brassicae Bezdenko and T. evanescens Westwood could be stored at
cold temperatures for various lengths of time without significant reductions on
parasitoid quality.
1Trakya University, Faculty of Agriculture, Department of Plant Protection,
Tekirdað, Turkey.  e-mail:nozder@tu.tzf.edu.tr.
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Ozder and Saglam: Effect of Short Term Cold Storage on the Quality of <i>Trichogram
Published by ValpoScholar, 2004
184 THE GREAT LAKES ENTOMOLOGIST Vol. 37, Nos. 3 & 4
MATERIALS AND METHODS
Parasitoids originally collected from Archips rosanus L.(Tortricidae) eggs
in Turkey were identified as T. cacoeciae (N. Kilinçer, University of Ankara,
Faculty of Agriculture, Department of Plant Protection) and reared on Ephestia
kuehniella Zeller (Pyralidae) since 1998.  T. brassicae and T. evanescens were
obtained from the Plant Protection Research Institute in Izmir and in Adana
(Turkey).  All Trichogramma species were reared on E. kuehniella in a climatic
chamber at 25 ±  1 °C, 60-70% R.H. and L16:D8.  E. kuehniella were reared on
wheat bran at 25 ± 1 °C and 60-70% R.H.
Twenty-four hour old eggs of E. kuehniella were sprinkled over a fine gum
film on paperboard strips, with 100 host eggs per strip.  The strips were placed
separately in glass vials (15 × 2.5cm) with ten newly emerged parasitoids and
held for one day at 25 ± 1 °C temperatures, 60-70% R.H and 16:8h (L:D) photo-
period.  After four days of parasitization (the parasitized eggs turn black as
parasitoids pupated), the strips were placed separately in glass vials (15 ×
2.5cm), plugged with cotton, and stored at 4 ± 1 °C, 65-70% R.H. with a complete
dark photoperiod for 1, 2, 3 and 4 weeks.  Parasitized eggs were removed every
week for all parasitoid species and incubated for emergence of parasitoids at
climatic conditions described above.  The number of adult parasitoids emerging
from host eggs was recorded.  All of the treatments at each storage period and
parasitoid species were replicated ten times.
Freshly emerged adults (< 12h old) were used for determining longevity
and fecundity of the adults emerging from pupae stored at cold temperatures.
The fecundity and longevity of parasitoids exposed to cold storage was deter-
mined using fresh eggs of E. kuehniella (< 12 h old).  For each parasitoid species
and for each replicate, a single fertilized and unfed female, less than 6 h after
adult emergence was confined in a glass vial (1.5 cm diameter by 3.5 cm height)
with an egg card that had > 50 host eggs.  The inside of the vial was streaked
with diluted 70% honey to provide a food source for the parasitoid.  After 24 h,
the egg card was removed and placed in a new vial which was incubated under
laboratory conditions (25 ± 1 °C, 60-70% RH and a photoperiod of L16:D8 ) until
emergence of the parasitoid offspring.  Fresh host eggs were provided daily; the
number of eggs offered was more than the maximum daily female capacity of
parasitism.  For each combination of parasitoids, the number of eggs parasit-
ized was determined by counting the number of eggs which had turned black on
the egg card.  After hatching of progeny, all black eggs were dissected to confirm
the hatching of a wasp or determine mortality based on dead Trichogramma
pupae.  As a control for this experiment, we used parasitoids which emerged
from host eggs held at the laboratory conditions (25 ± 1 °C, 60-70% RH and
L16:D8 light photoperiod).  Each treatment for all parasitoid species and tem-
perature regimes was replicated 10 times.  The effect of time and temperature
on the longevity and fecundity of parasitoids was analyzed using ANOVA. Means
were compared using Tukey’s Tests.
RESULTS AND DISCUSSION
Storage period had a significant effect on the emergence rate of adults of T.
cacoeciae (F = 18.89, df = 4, P < 0.05), T. evanescens (F = 23.59, df = 4, P < 0.05) and
T. brassicae (F = 10.20, df = 4, P < 0.05).  Emergence rates of adults from stored
hosts decreased with cold for all three parasitoid species compared to the control.
Development time from pupa to adult emergence was 5 days for all samples
in the control group and 6 days for all samples in each of the treatments.  These
results are similar to those for T. nerudai Pintureau and Gerding (Tezze and
Botto 2004).  While cold storage had little affect on development time it, af-
fected the proportion of emerged adults as well as their quality.
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The Great Lakes Entomologist, Vol. 37, No. 2 [2004], Art. 9
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2004 THE GREAT LAKES ENTOMOLOGIST 185
Emergence rates of T. cacoeciae which had been stored at 4 ± 1 °C for 0, 1,
2, 3 and 4 weeks were found to be 99.40%, 98.80%, 98.20%, 85.00% and 73.80%,
respectively.  Similarly, emergence rates for T. evanescens were 99.80%, 99.60%
98.20%, 89.60% and 85.60%, respectively for the same storage conditions.  T
brassicae emergence rates were found to be 100%, 99.33%, 99.20%, 85.80% and
85.40%, respectively for the same storage conditions (Table 1).  Emergence rates
decreased with increasing cold storage for T. cacoeciae (F = 18.89, df = 4, P <
0.05), T. evanescens (F = 23.59, df = 4, P < 0.05) and T. brassicae (F = 10.20, df =
4, P < 0.05).
A reduction in the emergence due to the cold storage was also seen in other
Trichogramma spp. by Jalali and Singh (1992), Pitcher et al. (2002).  Emergence
rates for T. cacoeciae, T. evanescens, and T. brassicae stored for 4 weeks at 4 ± 1°C
were similar to rates obtained for T. nerudai stored for 25 days at 4 °C (Tezze
and Botto 2004), but higher than rates determined by Jalalý and Singh (1992)
for T. chilonis Ishii and T. japonicum Ashmead stored at 5 °C for 28 days.
Longevity of emerging adult parasitoids decreased significantly with in-
creasing storage times for T. evanescens (F = 7.03, df = 4, P < 0.05) in comparison
to the control (Table 1).  This is similar to results found for T. ostriniae Pang and
Chen by Pitcher et al. (2002).  However, storage period was not found to affect
longevity of T. achaeae Nagaraja and Nagarkatti and T. chilonis by Jalali and
Singh (1992).  Likewise, duration of cold storage resulted in a significant de-
crease in fecundity for T. cacoeciae (F = 15.77, df = 4, P < 0.05), T. evanescens (F =
27.86,df = 4, P < 0.05), and T. brassicae (F = 3.63, df = 4, P < 0.05) (Table 1).  This
Table 1. Mean (± SE) emergence rates, longevity and fecundity of Trichogramma
cacoeciae, T. brassicae and T. evanescens held in cold storage for various lengths of
time (N=10 replicates).
Storage period Emergence Female longevity Female fecundity
(Weeks) rates (%) (days) (number of eggs)
Trichogramma cacoeciae
Control 99.40 ± 1.34 a 15.60 ± 1.14 a 65.33 ± 2.52 a
1 week 98.80 ± 0.84 a 15.25 ± 1.71 a 50.20 ± 1.92 a
2 weeks 98.20 ± 1.09 a 15.25 ± 10.81 a 49.20 ± 1.93 a
3 weeks 85.00 ± 3.32 b 10.80 ± 8.61 a 47.00 ± 8.92 a
4 weeks 73.80 ± 12.49 c  5.75 ± 2.22  a 13.33 ± 19.65 b
Trichogramma evanescens
Control 99.80 ±  0.45 a 21.25 ± 3.86 a 76.20 ± 15.07 a
1 week 99.60 ± 0.55 a 16.25 ± 5.91 ab 64.66 ± 1.53 a
2 weeks 98.20 ± 1.30 a 14.00 ± 9.84 ab 58.33 ± 5.13 a
3 weeks 89.60 ± 5.55 b 7.00 ± 4.97 b 30.75 ± 2.22 b
4 weeks 85.60 ± 3.58 b 4.60 ± 1.52 b 12.00 ± 10.00 b
Trichogramma brassicae
Control 100 ± 0.00 a 17.50 ± 9.85 a 74.67 ± 20.65 a
1 week 99.33 ± 1.15 a 16.67 ± 11.67 a 67.00 ± 22.60 a
2 weeks 99.20 ± 0.84 a 14.00 ± 7.94 a 49.20 ± 16.16 ab
3 weeks 85.80 ± 8.79 b 9.40 ± 8.68  a 32.40 ± 37.32 ab
4 weeks 85.40 ± 6.53 b 4.80 ± 4.09   a 8.33 ± 10.12 b
Within columns, the same letters indicate no significant differences between
treatments Tukey’s multiple comparison test, (P < 0.05).
3
Ozder and Saglam: Effect of Short Term Cold Storage on the Quality of <i>Trichogram
Published by ValpoScholar, 2004
186 THE GREAT LAKES ENTOMOLOGIST Vol. 37, Nos. 3 & 4
result agrees with results for T. ostriniae (Pitcher et al. 2002) and for T. nerudai
(Tezze and Botto 2004).  Others have found that fecundity by T. achaeae, T.
eldanae Viggiani, T. chilonis ,and T. japonicum declined rapidly after storage for
14 days at 2 ºC (Jalalý and Singh 1992).
According to this study, T. cacoeciae, T. evanescens, and T. brassicae pupae
are amenable to short term cold storage.  Longer storage periods caused a sig-
nificant decrease in percent emergence, longevity and fecundity of parasitoid
species.  Storage of T. cacoeciae, T. evanescens and T. brassicae pupae at low
temperature for short time could be useful for inundative biological control
strategies.  Moreover, since the quality of F1 longevity and fecundity were not
affected, this storage technique could also be useful in release programs since
these species are already adapted to the local environmental conditions.
LITERATURE CITED
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2004 THE GREAT LAKES ENTOMOLOGIST 187
Pitcher, S. A., M. P. Hoffmann, J. Gardner, M. G. Wright, and T. P. Kuhar.  2002.  Cold
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nerudai (Hymenoptera: Trichogrammatidae).  Biological Control 30: 11-16.
5
Ozder and Saglam: Effect of Short Term Cold Storage on the Quality of <i>Trichogram
Published by ValpoScholar, 2004


Effect of Short Term Cold Storage on the Quality of \u3ci\u3eTrichogramma Brassicae, T. Cacoeciae\u3c/i\u3e, and \u3ci\u3eT. Evanescens\u3c/i\u3e (Hymenoptera: Trichogrammatidae)

https://scholar.valpo.edu/cgi/viewcontent.cgi?article=2112&amp;context=tgle

Effect of Short Term Cold Storage on the Quality of \u3ci\u3eTrichogramma Brassicae, T. Cacoeciae\u3c/i\u3e, and \u3ci\u3eT. Evanescens\u3c/i\u3e (Hymenoptera: Trichogrammatidae)

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