Biological control of the South American Tomato Pinworm, Tuta

absoluta (Lep.: Gelechiidae), with releases of Trichogramma

achaeae (Hym.: Trichogrammatidae) on tomato greenhouse of
Spain

T. Cabello
1
, J.R. Gallego
1
, E. Vila
2
, A. Soler
2
, M. del Pino
3
, A. Carnero
3
, E.
Hernndez-Surez
3
, A. Polaszek
4

1
Dpto. Biologia Aplicada, Universidad de Almeria. Ctra. Sacramento s/n, 04120-
Almeria, Spain.E-mail: tcabello@ual.es;
2
Agrobo, Ctra Nacional 340, Km. 419,
04745-La Mojonera, Almera, Spain;
3
Instituto Canario de Investigaciones Agrarias
ICIA, Valle de Guerra, P.O. 60, 38200-La Laguna, Tenerife, Spain;
4
Department of
Entomology, Natural History Museum, Cromwell Road, London SW7 5BD, U.K.


Abstract: The egg parasitoid Trichogramma achaeae has been identified as a candidate for
biological control of the South American Tomato Pinworm, Tuta absoluta. On laboratory
conditions a marginal attack rate of 100 % was found and 83.3 % of the parasitized eggs
developed until the blackhead stage (apparent parasitism). On greenhouse conditions a high
efficacy (91.74 % of damage reduction) was obtained when releasing 30 adults/ plant (= 75
adults/ m
2
) every 3-4 days on August and September of 2008. This shows that this parasitoid
can be a good weapon to control T. absoluta on greenhouses of the southeast of Spain.

Key words: greenhouse, tomato, exotic pest species, oophagous parasitoid, biological
control, Tuta absoluta, Trichogramma achaeae.


Introduction

The South American Tomato Pinworm, Tuta absoluta (Meyrick), has spread from
Central America to most of South America, and it is considered one of the major pests
on tomato and other solanaceous crops (EPPO, 2005). Its chemical control is difficult,
firstly due to its biology, because of the mine-feeding behaviour of larvae (Branco. &
Franca, 1993; Urbaneja et al., 2007), which has originated an intensive use of chemical
treatments. Secondly, it has developed insecticide resistance to the active ingredients
used until now (Siqueira et al., 2000; EPPO, 2005; Lietti et al., 2005). Consequently,
biological control has been developed and it is being applied on several South American
countries, especially using the oophagous parasitoids Trichogramma pretiosum (Riley)
and T. nerudai Pintureau and Gerding (Riquelme & Botto, 2003; Parra & Zucchim,
2004; Bueno, 2005; Faria et al., 2008).
T. absoluta was accidentally introduced in Castelln, Spain, in 2006; it has spread
from Comunidad Valenciana to Murcia and Ibiza (Urbaneja et al., 2007; EPPO, 2008)
and first infestations were detected in the greenhouses of Almeria in 2008. This
quarantine pest has caused severity damage on tomato crops in Murcia area (Lucas et
al., 2009), although impact is still fairly low in Andaluca (Gonzalez-Garcia, 2009).
Several species of autochthonous predators have been evaluated as biological control
agents in Spain with promising results, mainly the mirid bugs Nesidiocoris tenuis
(Reuter) and Macrolophus pygmaeus (Rambur) (Urbaneja et al., 2008) and the damsel
bug Nabis pseudoferus Remane (Cabello, 2009). However, the establishment of these
predators into the crops is often too slow to avoid damage of the pest, and consequently
chemical treatments are needed, which disturbs biological control. Then, several works
are being conducted to evaluate autochthonous parasitoids, using them alone or
combined with predators, on tomato greenhouse crops from the southeast of Spain.
Preliminary results with the specie Trichogramma achaeae Nagaraja & Nagarkatti are
presented on this work.
T. achaeae has a world wide distribution; it has been mentioned in Asia (China,
India and Russia), Europe (France, Russia), Africa (Cabo Verde) and the New World
(Argentina, Barbados, Chile, Trinidad & Tobago, U.S.A.). Recently, it has been
reported in Spain (unpublished data). It is an egg parasitoid of 26 Lepidoptera species
belonging to 10 families: Gelechiidae, Geometridae, Noctuidae, Notodontidae,
Oecophoridae, Pieridae, Pyralidae, Sphingidae, Tortricidae and Yponomeutidae
(Polaszek, not publ.). T. achaeae has been evaluated as a biological control agent of
different lepidopteran pests (Jalali et al., 2001; Jalali & Singh, 2002; Jalali et al., 2002;
Chandrashekhar et al., 2003; Yadav & Anand, 2003) and is commercially available in
India (Nagaraja et al., 2002).


Material and methods

Insects rearing
The individuals of T. absoluta used in the trials were obtained from a laboratory rearing
on tomato leaves. The rearing of T. achaea, also in the laboratory, was conducted using
eggs of Ephestia kuehniella Zeller as alternative host, according to the methodology
described by Cabello (1985).
Laboratory bioassay
A preliminary test under laboratory conditions (251 C, 60-80 % RH and 16:8 hours
L:D) was carried out to evaluate acceptance and parasitation of T. absoluta eggs by
females of the parasitoid. Methodology was adapted from Brotodjojo & Walter (2006),
with the exception that 10 eggs of T. absoluta (less than 24 hours aged) were offered to
each female. 10 repetitions were conducted. All the parasitized eggs were developed
until adult emergence.
Greenhouse bioassay
To evaluate the efficacy of T. achaea a test was conducted in a commercial greenhouse
of Almeria (Spain), between August the 27
th
and September the 22
th
of 2008. Eight
cages (8 m
2
) were placed into the greenhouse with 20 tomato plants (Cherry variety) per
cage arranged on pots. Plants were infested with T. absoluta (4 adults/ plant) when
reaching one meter height.
A total of seven releases of T. achaeae (30 adults/ plant) were conducted every
three or four days on four cages. The other four cages were leaved as a control. The
number of larves of T. absoluta, leaf mines and damaged fruits were counted alter 27
days. Data was analyzed using one-way ANOVA and means were compared with LSD,
SPSS software v. 15 (SPSS, 2006).


Results and discussion

Laboratory bioassay
The eggs of T. absoluta were well accepted as hosts by T. achaea (100 % of the eggs
received a laying of the parasitoid). However, only 83.3 % of the eggs developed to the
blackhead stage (prepupa of the parasitoid) (apparent parasitism), while the rest did not
and finally collapsed. The eggs of T. absoluta are smaller than the eggs of the rearing
host, E. kuehniella. Adult females of T. achaeae, both reared on Chrysodeixis chalcites
(Esper) or E. kuehniella, are larger than the eggs of Tuta absoluta but shorter than the
eggs of E. kuehniella (fig.1). It has been described that the size of the rearing host does
affect the parasitism rate of natural hosts (Brotodjojo & Walter, 2006); female
parasitoids in these cases accept host eggs with about the same size as their natal host or
larger ones (Salt, 1940; Nurindah et al., 1999). In the present work T. achaeae accepted
all the offered eggs of T. absoluta, although the small size of the host may have affected
the development of the parasitoid, which could explain why a percentage of the eggs
collapsed.
Adults of Trichogramma also kill host eggs by feeding on them; the host egg is
stung and the female feeds on the drop of liquid appearing at the site of the sting. The
host egg dies, leaving no evidence of parasitism (Knutson, 1998). In some species of
Trichogramma, host feeding contributes significantly to pest control (Vasquez et al.,
1997). In the present work host feeding by female parasitoids was not observed.
However, all the individuals had a supplier of food, a drop of honey:water (1:1), during
the bioassay, and host feeding could happen under field conditions.


T. absoluta
egg E. kuehhniella
egg
T. achaeae (from Ch.
chalcites egg) T. achaeae (from E.
kuehniella egg)
Population
0
200
400
600
A
v
e
r
a
g
e

(

m
)
Length
Width


Figure 1. Comparative size of E. kuehniella eggs, T. absoluta eggs and adult females of
the parasitoid (T. achaeae) when reared on eggs of Chryxodeixis chalcites or Ephestia
kuehniella (C.L. at P=0.05).


This means that checking the colour as a way to evaluate the ratio of parasitism,
both on field or laboratory conditions, can underestimate the real impact. For this
reason, egg hatch should be recorded in addition to egg parasitism (Knutson, 1998).
It could also explain why data about the rate of parasitism, in this case of T.
pretiosum, shows low values, among 1.5 % (Faria et al., 2008), 22.7-24.4 % (Villas-
Boas & Franca, 1996) and 49.0 % as the maximum (Haji et al., 1995), although it is a
real effective field method to control the pest on several countries of South American.
Then, it is a subject to be considered when establishing the methodology to estimate the
efficacy of Trichogramma to control this pest on field conditions.

Greenhouse bioassay
On figure 2 the number of larvae, leaf mines and damaged fruits on tomato plants after
27 days from the beginning of the trial are showed. Number of larvae and damaged
leaves are significantly lower where parasitoids had been released (P < 0.01) and the
numbers of damaged fruits is also lower (P < 0.05) on this treatment. The number of
larves of T. absoluta was 12 times lower on plots where T. achaeae was released
compared with the control. This means a 91.74 % of efficacy.
On one hand, the doses of releases used in this bioassay are similar to that used
commercially in Brasil with T. pretiosum (Freitas et al., 1994: Parra & Zucchim, 2004).
Likewise, the levels of damaged fruits are similar to that described when using T.
pretiosum, with cited values from 2.0 % (Villas-Boas & Franca, 1996) to 13.0 %
(Domingues et al., 2003). On the other hand, the determined efficacy (91.7 %) is lightly
higher than the cited when using T. pretiosum (87.7 %) on said country (Parra &
Zucchim, 2004).


4.95a
0.42b
23.5a
5.86b
0.85a
0.18b
Control T. achaeae releases
Treatment
0
5
10
15
20
25
N
u
m
b
e
r

p
e
r

p
l
a
n
t
Larvae
Leaf miners
Damaged fruits


Figure 2. Number of Tuta absoluta larvae and damage on tomato plants counted in a
greenhouse according to the treatment (with or without releases of T. achaeae).


Showed data are promising, but must be completed with other studies (now in
course), mainly to establish the needed releases on greenhouses according to the density
of the pest and the growth stage of the crop, in the protected crops of the southeast of
Spain. A good possibility may be to combine releases of T. achaeae with the mirid
predator N. tenuis. One of the major problems of using N. tenuis in Almeria is his
slowly establishment, needing at least around 4 weeks. Tomato crop is mainly planted at
the end of the summer, so releases are made in autumn, when temperature is dropping.
During this period Tuta absoluta can cause heavy damage, and weekly releases of the
parasitoid could help to guaranty an effective control of the pest.


Acknowledgements

This publication has been realized with the support of the Ministry of Sciences and
Innovation (of Spain): CDTI Programme, CENIT-MEDIODIA Project.

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