J. Agr. Sci. Tech. (2015) Vol. 17: 1507-1516

Cold-Storage of Piezodorus guildinii (: ) Eggs for Rearing Telenomus podisi (Hymenoptera: )

M. Fernanda Cingolani 1∗, N. M. Greco 1, and G. G. Liljesthröm 1

ABSTRACT

Piezodorus guildinii (Hemiptera: Pentatomidae) is an important soybean pest, and one of its main natural enemies is Telenomus podisi (Hymenoptera: Platygastridae). Rearing of the parasitoid is constrained by the hosts' egg quality, which deteriorates after few generations in laboratory, therefore, cold-stored host eggs utilization could be a useful tool for augmentative biological control. Thus, the objective was to evaluate the quality of P. guildinii cold-stored eggs, on the performance of parental and F 1 generation of T. podisi . Host eggs 24 hour old were stored at -18°C for one (N= 53), two (N= 28) or three months (N= 29), and approximately 40 host eggs were offered to a T. podisi female per treatment, for 48 hours. The control treatment consisted of 24-hour-old non-frozen host eggs, obtained and kept at 24°C (N= 55). Parental generation parasitism and progeny´s emergence on frozen eggs was significantly lower than on non-frozen eggs, even for the shorter storage period. Male proportion and preimaginal development time of the progeny increased with freezing period. However, parasitism proportion caused by adults of F 1, and emergence, male proportion, and preimaginal development time of F 2 were not affected. Although the performance of T. podisi on frozen P. guildinii eggs was significantly lower than on nonfrozen ones, host eggs storage for up to two months allowed obtaining a parasitism rate of 40% with a high emergence rate. This could be helpful enough to maintain mass rearings, mainly during the host hibernation period, and to enhance field parasitism when host is scarce.

Keywords : Host storage, Mass-rearing, Parasitoids.

INTRODUCTION susceptible than other bugs to labeled insecticides such as pyrethroids and Phytophagous hemipterans of the family organophosphates (Temple et al. , 2013a). It Pentatomidae, commonly known as stink is one of the most predominant stink bugs bugs, are important soybean pests. Among and a serious pest of soybean in the south them, Piezodorus guildinii Westwood Nearctic and whole Neotropical regions (Hemiptera: Pentatomidae) is the one that (Castiglioni et al. , 2010; Corrêa-Ferreira, most affects the quality and viability of 2008; Massoni and Frana, 2006; Temple et seeds and causes greater leaf retention when al. , 2013b). In the last two decades P. compared to other common bugs like guildinii relative abundance has been Nezara viridula L. and Euschistus heros (F.) increasing significantly and has currently (Hemiptera: Pentatomidae) (Husch et al. , become the most important species in 2014). Piezodorus guildinii is a Neotropical Buenos Aires and other provinces of multivoltine species, highly mobile and Argentina (Cingolani, 2012; Cingolani et al. , more difficult to control as it is less 2014). ______1 CEPAVE (CCT La Plata CONICET-UNLP), Boulevard 120 between 60 and 64 - La Plata (1900), Buenos Aires, Argentina. ∗ Corresponding author; e-mail: [email protected]

1507 ______Fernanda Cingolani et al.

In the Neotropical region the most and Foerster, 2013; Favetti et al. , 2014; common egg parasitoid species of P. Kivan and Kilic, 2005; Mahmoud and Lim, guildinii are Telenomus podisi (Ashmead), 2007). Trissolcus urichi (Crawford) and Trissolcus Several intrinsic platygastrids basalis (Wollaston) (Hymenoptera: characteristics, such as simplicity of adults’ Platygastridae) (Castiglioni et al. , 2010; diet, lack of hyperparasitoids and pathogens, Cingolani et al. , 2014; Corrêa-Ferreira, good reproductive capacity, and small size 2005; Molinari, 2005). Over 20 species of (and therefore minimum space platygastrid wasps have been used for requirements) make the rearing of these biological control of stink bugs in several wasps feasible. However, supporting large countries (Luck, 1981; van Lenteren and colonies of these parasitoids is strongly Bueno, 2003). Trissolcus basalis releases constrained by the quality of their hosts, mainly to control N. viridula have been which deteriorates after few generations made in Argentina (Crouzel and Saini, with currently used laboratory techniques 1983), Brazil (Corrêa-Ferreira and (Parra and Cônsoli, 2009). For the specific Moscardi, 1995; Corrêa-Ferreira and case of P. guidinii , the establishment of Moscardi, 1996), Italy (Colazza and Bin, large colonies is quite difficult due to the 1995) and the United States (Hoffmann et fact that very few individuals, obtained from al. , 1991), and to control this stink bug and eggs collected from field, can reach the adult Agonoscelis glitters F. (Hemiptera: stage. In addition, reproduction begins just Pentatomidae) in Australia (Clarke, 1990). after the sixth week of life (15 days after the Telenomus podisi has been used to control imaginal molt), only 60 or 70% of the E. heros , P. guildinii and N. viridula in females are fertile (Panizzi and Slansky, soybean organic production fields in Brazil 1985), and each of them performs only three (Sujii et al. , 2002). to five egg masses of on average of 14 eggs A frequent difficulty when implementing each, during its lifetime. Another important augmentative biological control is to obtain factor is the high nymphal mortality (60%) large numbers of control agents of that is registered with the commonly used appropriate quality, at the time they are rearing techniques (Serra and La Porta, required (Orr, 1988). Various techniques 2001) and the difficulty of maintaining the have been developed to optimize the colony throughout the year, because even maintenance of large parasitoids' rearing in under optimal temperature and photoperiod the laboratory. Some of them are focused on conditions, reproduction is interrupted each parasitoids, such as cold storage of year due to hibernation, at least in the parasitized hosts (Bayram et al ., 2005; Dass geographic region between 30°-35° S and Ram, 1983; Gautam, 1986; Noble, latitude (Zerbino et al. , 2013). 1937), in vitro development of parasitoids There is evidence that cold storage of host (Shirazi, 2006), and cold storage of pupae or eggs can have positive, negative or neutral adults of the parasitoid (Bayram et al ., 2005; effects on the performance of platygastrids Foerster et al. , 2004; Foerster and Doetzer, parasitoids (Orr, 1988 and references 2006; Gautam, 1986). Other techniques therein), and no adverse effects were found focus on ensuring a constant availability of in F 1 generation (Alim and Lim, 2010; enough host, such as the development of Mahmoud and Lim, 2007). The objective of artificial diets (Fortes et al. , 2006; Panizzi et this study was to evaluate the quality of P. al. , 2000), irradiation of host eggs with guildinii eggs preserved at low temperatures gamma ( γ) rays to increase the time during for different storage periods and to assess which they are likely to be parasitized the parasitism rate, and progeny's (Nordlund et al. , 1983) and cold storage of emergence, sex ratio and preimaginal healthy (not parasitized) hosts (Alim and development time, of parental and F 1 Lim, 2010; Corrêa-Ferreira, 1998; Doetzer generations.

1508 Cold Storage of Piezodorus guildinii Eggs ______

emerged after 12 days i.e. average preimaginal parasitoid development time at MATERIALS AND METHODS 24°C, (Corrêa-Ferreira, 1993) were dissected a week later to check presence of pupa or dead adult parasitoids inside the and Rearing Procedures host. We evaluated parasitism by parental Colonies of stink bugs and wasps were generation, and emergence, male proportion, established from field collected individuals. and preimaginal development time of F 1 on Adults of P. guildinii were fed Phaseolus nonfrozen and frozen eggs. Additionally, we vulgaris (L.) pods replaced every two days, evaluated parasitism by F 1 and emergence, male proportion and preimaginal and maintained in cages (15x15x30cm) at nd 24±1°C, 70±10% RH and 14:10 L:D. development time of F 2 (2 filial Deposited eggs were collected daily, and T. generation). podisi was reared on these eggs, and kept in test tubes under the same laboratory Statistical Analysis conditions with honey as food source. All parasitoids females used in experiments were inexperienced, two-days old and fed Parasitism rate (number of eggs from ad-libitum. which a pupa or an adult parasitoid was observed/total number of offered eggs) and emergence proportion (number of emerged Cold Storage Treatments wasps/number of eggs from which a pupa or an adult parasitoid was observed) were Piezodorus guildinii eggs 24-hour-old compared among treatments, using the were stored at -18°C for one, two, or three Kruskal-Wallis test (P< 0.05). We months (treatments) wrapped in aluminum performed the Kruskal-Wallis H test only foil, following the methodology proposed by when ANOVA assumptions were not met, Corrêa-Ferreira and Moscardi (1993). After even after applying transformations to our the storage period, eggs were kept at 5°C for data. We counted the total number of two hours and then at 24°C for another two females and males developed in each 2 hours for acclimation of stored eggs before treatment. We used χ of 2×4 contingency being offered to the parasitoids table to analyze if the number of adults of (Albuquerque et al. , 2000). On each both sexes among treatments differed from treatment, a group of approximately 40 host what could be expected by chance, and eggs were offered to T. podisi females. subdivided contingency tables were Fifty-three replicates for one month stored analyzed to find significant differences eggs, 28 replicates for two months stored between pairs of treatments (Zar, 1996). We eggs, and 29 replicates for three months also compared female progeny’s stored eggs were performed. In order to preimaginal development time (from the avoid pseudoreplication, replicates of each beginning of the experiment to emergence) treatment periods were performed starting at among treatments, using the Kruskal-Wallis different times over the year. The control H test. treatment consisted of 24-hour-old In addition, the effect of one month host nonfrozen host eggs, obtained and kept at egg refrigeration on F 1 generation of the 24°C (N= 55). Eggs were exposed to one parasitoid was assessed. Forty T. podisi female parasitoid for 48 h and then were females randomly selected among the group kept under controlled conditions (24±1°C, of adult parasitoids emerged from one 70±5% RH, 14:10 L:D) until emergence of month stored host eggs, and 55 T. podisi parasitoids. Eggs from which no parasitoid females selected among group of adult

1509 ______Fernanda Cingolani et al. parasitoids emerged from nonfrozen eggs differed among treatments ( χ2= 28.925, df= were provided with approximately 40 3, P= 0.05), and only in the three months nonfrozen eggs of P. guildinii for 48 h in frozen eggs, the sex ratio was different, as controlled conditions (24±1°C, 70±5% RH, suggested by a subdivided contingency table 14 hours photophase). Proportion parasitism, considering three months frozen eggs versus emergence rate, and preimaginal all other treatments ( χ2= 23.865, df= 2, P= development time were compared using 0.05) (Table 1). ANOVA. Sex ratio was analyzed using χ2 of Preimaginal development time of the a 2×2 contingency table. female progeny of the parental generation was significantly longer in wasps developed RESULTS from one month frozen eggs (H (3, N=107)= 26.796, P< 0.001) (Table 1). In every case, males emerged the day before females’ No host nymphal development was emergence date. recorded from frozen eggs. Parasitism rate For the F 1, parasitism rate caused by T. by parental generation was significantly podisi females emerged from one month lower on frozen host eggs than on nonfrozen frozen eggs was similar to that of females ones (H (3, N=165)= 71.060; P< 0.001), and the emerging from nonfrozen eggs (F= 1.720, lowest value was recorded for the storage df= 1, P= 0.193) (Figure 2). Emergence rate period of three months (Figure 1). Similarly, (F= 0.74, df= 1, P= 0.393) (Figure 2), sex the progeny of the parental generation had a ratio ( χ2= 3.059, df= 1, P= 0.08) and significantly lower emergence proportion preimaginal development time of the than those developed in nonfrozen eggs (H (3, progeny of F 1 generation (i.e. F 2) (F= 0.24, N=135)= 36.299, P< 0.001) (Figure 1). Also, df= 1, P= 0.625) (Table 2) were also similar the parental generation progeny´s sex ratio between storage treatments.

Figure 1. Parasitism and progeny's emergence proportion of the parental generation of Telenomus podisi , from nonfrozen or frozen Piezodorus guildinii eggs. Bars indicate the confidence interval at 95%. Different letters indicate significant differences (by Kruskal-Wallis) on parasitism (upper-case letters) and emergence (lower-case letters) among non-frozen and frozen eggs.

1510 Cold Storage of Piezodorus guildinii Eggs ______

Table 1. Progeny's male proportion [Male/(Male+Female)] and females' preimaginal development time (days) (mean±SE) of the parental generation of T. podisi developed in nonfrozen and frozen eggs of P. guildinii stored during one, two or three months. a Progeny's male Females' preimaginal Treatment proportion development time 0.126±0.008a 12.982±0.057a Nonfrozen (n= 55) (n= 55) 0.091±0.011a 15.125±0.206b 1 month frozen (n=32) (n= 32) 0.084±0.014a 13.125±0.189a 2 months frozen (n= 26) (n= 16) 0.250±0.015b 12.500±0.147a 3 months frozen (n= 16) (n= 4) a Numbers in each column followed by a different letter are significantly different (P< 0.05) for contingency table (progeny's male proportion) or Kruskal-Wallis.

Table 2. Progeny's male proportion [Male/(Male+Female)] and females' preimaginal st development time (days) (mean±SE) of the parental generation and F 1 (1 filial generation) of T. podisi developed in non-frozen P. guildinii eggs. a Parasitoid Progeny's male Females' preimaginal generation proportion ns development time ns 0.132±0.007 12.982±;0.057 Parental (df= 54) (df= 54) 0.106±0.007 13.075±0.083 F 1 (df= 39) (df= 39) a ns: Non significantly different (P< 0.05) for contingency table.

st Figure 2. Parasitism and progeny's emergence proportion of the parental generation and F 1 (1 filial generation) of Telenomus podisi in nonfrozen Piezodorus guildinii eggs. Bars indicate the confidence interval at 95%.

1511 ______Fernanda Cingolani et al.

DISCUSSION provoked changes in the hardness of the host egg chorion (Kivan and Kilic, 2005), making it Mass-rearings of platygastrid parasitoids more difficult for the wasps to cut it with their demand large numbers of host eggs, as it has mouth parts for emergence. not been possible to maintain platygastrids The proportion of female wasps developed species colonies without rearing the host up to in frozen host eggs stored for three months the present (Orr, 1988 and references therein; was the lowest. As usually happens with Shirazi, 2006; Strand et al. , 1988; Volkoff et hymenopteran parasitoids, T. podisi have a al. , 1992). Besides, stink bugs rearing are haplo-diploid sex-determination system, constrained, as they have a poorer female being able to choose the sex of their development on artificial diets (Fortes et al. , progeny by controlling fertilization (Flanders, 2006; Silva and Panizzi, 2008). In this sense, 1946). Distortions of sex ratio may come from cold storage of host eggs seems to be a modification of the proportion of fertilized interesting to obtain large numbers of control eggs oviposited i.e. primary sex ratio, or from agents of appropriate quality, at the time they a differential mortality between sexes i.e. are required. secondary sex ratio. The sex ratio found in this Our results showed that P. guildinii eggs study would indicate that an increase in storage for up to two months allowed storage time of frozen eggs made host quality obtaining an acceptable parasitism rate by T. to diminish, pushing females to oviposit more podisi with high emergence. Parasitism on male progeny. frozen host eggs was significantly lower than The longer preimaginal development time that observed on nonfrozen eggs, and had needed by wasps to complete their halved when eggs were preserved in freezer development on one month frozen eggs also for one month. Other authors have shown that denotes a decline in hosts' quality. In this parasitism and progeny's emergence from cold study, only a low number of adults could stored hosts were variable with different emerge from eggs stored for longer period temperatures and storage periods (Corrêa- (three months), and 20% of the parasitoids Ferreira and Moscardi, 1993; Orr, 1988). died at pupal stage. This may suggest that the Nezara viridula eggs preserved at low quality of this host would have prevented temperatures between -2 and -10°C remained wasps to complete their development. viable for parasitism by T. basalis for less than Parasitism rate by the F 1 and emergence of 60 days (Albuquerque et al. , 2000) while if the F2 from host eggs frozen for one month was preservation of eggs occurred in freezer, as in not affected. Similarly, other authors did not this study, parasitoid development was not find negative effects of pentatomids' eggs cold affected until a storage period of 150 days. In storage on platygastrids' second generation contrast, Dolycoris baccarum (L.) (Hemiptera: (Alim and Lim, 2010; Mahmoud and Lim, Pentatomidae) freezer stored eggs have 2007). remained viable for Trissolcus nigripedius Although the performance of T. podisi on Ashmead (Hymenoptera: Platygastridae) for frozen P. guildinii eggs was significantly only 8 days (Mahmoud and Lim, 2007). lower than on non-frozen eggs, storage for up Telenomus podisi progeny's emergence was to two months allowed obtaining a parasitism lower from frozen eggs than from nonfrozen rate of 40% with an emergence rate of almost ones. We observed many parasitoid pupae and 80%. This could be helpful enough to maintain adults dead inside the host egg. This may mass rearing mainly during the host indicate a decrease in egg quality as a hibernation period, or to face adverse resource, which could have diminished larval circumstances that can cause development and/or the capacity of developed unsynchronization of host-parasitoids rearing. adults to emerge. On the other hand, freezing An important technique for maintaining and subsequent thawing could also have suitable host for longer periods is the storage of host eggs in liquid nitrogen. For example,

1512 Cold Storage of Piezodorus guildinii Eggs ______

Graphosoma lineatum (L.) (Hemiptera: Traps Increase Field Parasitism in Soybean. J. Pentatomidae) eggs remained suitable for Econ. Entomol., 104 : 1833-1839. Trissolcus simoni (Mayr) (Hymenoptera: 3. Albuquerque, F. A., de Sousa Lobo Lima, S., Platygastridae) for as long as five years Zabini, A. V., Pattaro, F. C. and Maestro (Gennadiev and Khlistovskii, 1980). Even Borges, L. 2000. Viabilidade de Ovos de Nezara viridula (L.) Armazenados a Baixas though, when a short storage period is Temperaturas para o Parasitismo por sufficient, the use of frozen host eggs could be Trissolcus basalis (Woll.). Acta Sci., 22 : 963- a cheaper and handier technique than 967. maintaining hosts in liquid nitrogen. 4. Bayram, A., Ozcan, H. and Kornosor, S. 2005. Interestingly, cold-stored host eggs could Effect of Cold Storage on the Performance of also be useful to build up the population of Telenomus busseolae Gahan (Hymenoptera: natural enemies in the field by mass releasing Scelionidae), an Egg Parasitoid of Sesamia non-viable host eggs when hosts is limiting in nonagrioides (Lefebvre) (Lepidoptera: Noctuidae). Biol. Control, 35 : 68-77. the field (Leopold, 1998). Lim and Mahmoud (2009), Alim and Lim (2011) and Mainali et 5. Castiglioni, E., Ribeiro, A., Alzugaray, R., Silvam H., Ávila, I. and Loiáconom M. S. al . (2012) have demonstrated that this tool 2010. Prospección de Parasitoides de Huevos could help to increase platygastrids field de Piezodorus guildinii (Westwood) parasitism. (Hemiptera: Pentatomidae) en el Litoral Oeste de Uruguay. Agrociencia Uruguay, 14 : 22-25.

AKNOWLEDGEMENTS 6. Cingolani, M. F. 2012. Parasitismo de Huevos de Piezodorus guildinii (Hemiptera: Pentatomidae) por Trissolcus basalis y This study was supported by the National Telenomus podisi (Hymenoptera: Scelionidae) Agency for Promotion of Science and en el Noreste de la Provincia de Buenos Aires. Technology of Argentina (Agencia Nacional PhD. Dissertation, Facultad de Ciencias de Promoción Científica y Tecnológica), grant Naturales y Museo (UNLP), Argentina. 7. Cingolani, M. F., Greco, N. M. and PICT 15150, and by the Program of Incentive Liljesthröm, G. G. 2014. Egg Parasitism of for Professors-Researchers of the National Piezodorus guildinii and Nezara viridula Ministry of Culture and Education of (Hemiptera: Pentatomidae) in Soybean, Alfalfa Argentina (Programa de Incentivos a and Red Clover. Revista de la Facultad de Docentes-Investigadores del Ministerio de Ciencias Agrarias, UNCuyo, 46 : 15-27. Cultura y Educación de la Nación), grants 8. Clarke, A. R. 1990. The Control of Nezara 11/N454, 11/N 579 and 11/N 658. viridula L. with Introduced Egg Parasitoids in MFC was supported by a scholarship from Australia. A Review of a "Landmark" the Commission of Scientific Research from Example of Classical Biological Control. Aust. the province of Buenos Aires (Argentina) J. Agr. Res., 41 : 1127-1146. 9. Colazza, S. and Bin, F. 1995. Efficiency of (CIC) and the National Council for Scientific Trissolcus basalis (Hymenoptera: Scelionidae) and Technological Research (CONICET). as an Egg Parasitoid of Nezara viridula (Heteroptera: Pentatomidae) in Central Italy. REFERENCES Environ. Entomol., 24 : 1703-1707. 10. Corrêa-Ferreira, B. S. 1993. Utilização do Parasitóide Trissolcus basalis (Wollaston) No 1. Alim, M. A. and Lim, U. T. 2010. Biological Controle de Percevejos da Soja. Embrapa, Attributes of Ooencyrtus nezarae Ishii Londrina, Brazil, 40 PP. (Hymenoptera: Encyrtidae) Reared on 11. Corrêa-Ferreira, B. S. 1998. Viability of Refrigerated Eggs of Riptortus pedestris (= Nezara viridula (L.) Eggs for Parasitism by clavatus ) Fabricius (Hemiptera: Alydidae). J. Trissolcus basalis (Woll.), under Different Asia-Pacific Entomol., 13: 139-143. Storage Techniques in Liquid Nitrogen. Anais 2. Alim, M. A. and Lim, U.T. 2011. Refrigerated Sociedade Entomológica do Brasil, 27 : 101- Eggs of Riptortus pedestris (Hemiptera: 107. Alydidae) Added to Aggregation Pheromone

1513 ______Fernanda Cingolani et al.

12. Corrêa-Ferreira, B. S. 2005. Suscetibilidade da Ashmed (Hymenoptera: Scelionidae). Biol. Soja a Percevejos na Fase Anterior ao Control , 36 : 232-237. Desenvolvimiento das Vagens. Pesqui. 23. Foerster, L. A., Doetzer, A. K. and Ferreira de Agropecu. Bras., 40 : 1067-1072. Castro, L. C. 2004. Emergence, Longevity and 13. Corrêa-Ferreira, B. S. 2008. Trissolcus basalis Fecundity of Trissolcus basalis and Telenomus Para o Controle de Percevejos da Soja. In: podisi after Cold Storage in the Pupal Stage. “Controle Biológico no Brasil: Parasitóides e Pesqui. Agropecu. Bras., 39 : 841-845. Predadores ”, (Eds.): Parra, J. R. P., Botelho, P. 24. Fortes, P., Magro, S. R., Panizzi, A. R. and S. M., Corrêa-Ferreira, B. S. and Bento, J. M. Parra, J. R. P. 2006. Development of a Dry S.. Mandole, Piracicaba, Brazil, PP. 449-471. Artificial Diet for Nezara viridula (L.) and 14. Corrêa-Ferreira, B. S. and Moscardi, F. 1993. Euschistus heros (Fabricius) (Heteroptera: Técnicas de Armazenamento de Ovos do Pentatomidae). Neotrop. Entomol., 35 : 567- Percevejo-verde Visando à Multiplicação do 572. Parasitoide Trissolcus basalis (Wollaston). 25. Gautam, R. D. 1986: Effect of Cold Storage on Pesqui. Agropecu. Bras., 28 : 1247-1253. the Adult Parasitoid Telenomus remus Nixon 15. Corrêa-Ferreira, B. S. and Moscardi, F. 1995. Scelionidae: Hymenoptera and the Parasitised Seasonal Occurrence and Host Spectrum of Eggs of Spodoptera litura Fabr. Noctuidae Egg Parasitoids Associated with Soybean Lepidoptera. J. Entomol. Res., 10 : 125-131 Stink Bugs. Biol. Control, 5: 196-202. 26. Gennadiev, V. G. and Khlistovskii, E. D. 1980. 16. Corrêa-Ferreira, B. S. and Moscardi, F. 1996. Long Term Cold Storage of Host Eggs and Biological Control of Soybean Stink Bugs by Reproduction in Them of Egg Parasites of Inoculative Releases of Trissolcus basalis . Pest. Zh. Obshch. Biol., 41 : 314-319. Entomol. Exp. Appl., 79 : 1-7. 27. Hoffmann, M. P., Davidson, N. A., Wilson, L. 17. Crouzel, I. S. and Saini, E. D. 1983. T., Ehler, L. E., Jones, W. A. and Zalom, F. G. Importación de Trissolcus basalis (Wollaston) 1991. Imported Wasp Helps Control Southern (Hym. Scelionidae) en la Argentina para el . Calif. Agr., 45 : 20-22. Control Biológico de Nezara viridula (L. 28. Husch, P. E., de Olivera, M. C. N. and Sosa- (Hem. Pentatomidae). Revista de la Sociedad Gómez, D. R. 2014. Characterization of Injury Entomológica Argentina , 42 : 257-260. Caused by Edessa meditabunda (F.), Chinavia 18. Dass, R. and Ram, A. 1983. Effect of Frozen impicticornis (Stål), and Piezodorus guildinii Eggs of Corcyra cepahlonica Staiton (West.) (Hemiptera: Pentatomidae) to (Pyralidae: Lepidoptera) on Parasitism on Soybean. Neotrop. Entomol. , 43: 276-281. Trichogramma exiguum (Pinto and Platner) 29. Kivan, M. and Kilic, N. 2005. Effects of (Trichogrammatidae: Hymenoptera). Indian J. Storage at Low-temperatures of Various Entomol., 45 : 345-347. Heteropteran Host Eggs on the Egg Parasitoid, 19. Doetzer. A. K. and Foerster, L. A. 2013. Trissolcus semistriatus . BioControl, 50 : 589- Storage of Pentatomid Eggs in Liquid Nitrogen 600. and Dormancy of Trissolcus basalis 30. Leopold, R. A. 1998. Cold Storage of Insects (Wollaston) and Telenomus podisi Ashmead for Integrated Pest Management. In: (Hymenoptera: Platygastridae) Adults as a “Temperature Sensitivity in Insects and Method of Mass Production. Neotrop. Application in Integrated Pest Management ”, Entomol., 42 : 534-538. (Eds.): Hallman, G. J. and Denlinger, D. L.. 20. Favetti, B. M., Butnariu, A. R. and Doetzer, A. Westview Press, Oxford, UK, PP. 235-267. K. 2014. Storage of Euschistus heros Eggs 31. Lim, U. T. and Mahmoud, A. M. A. 2009. (Fabricius) (Hemiptera: Pentatomidae) in Inoculation of Refrigerated Non-viable Eggs Liquid Nitrogen for Parasitization by of Riptortus clavatus (Heteroptera: Alydidae) Telenomus podisi Ashmead (Hymenoptera: to Enhance Parasitism by Egg Parasitoids in Platygastridae). Neotrop. Entomol., 43 : 291- Soybean Field. Appl. Entomol. Zool., 44 : 37- 293. 45. 21. Flanders, S. E. 1946. Control of Sex and Sex- 32. Luck, R. F. 1981. Parasitic Insects Introduced limited Polymorphism in the Hymenoptera. Q. as Biological Control Agents for Rev. Biol., 21 : 135-143. Pests. In: “ Handbook of Pest Management in 22. Foerster, L. A. and Doetzer, A. K. 2006. Cold Agriculture ”, (Ed.): Pimentel, D.. CRC Press, Storage of the Egg Parasitoids Trissolcus Boca Ratón, USA, PP. 125-284. basalis (Wollaston) and Telenomus podisi

1514 Cold Storage of Piezodorus guildinii Eggs ______

33. Mahmoud, A. M. A. and Lim, U. T. 2007. 44. Shirazi, J. 2006. Investigation on the In vitro Evaluation of Cold-stored Eggs of Dolycoris Rearing of Trissolcus grandis an Egg baccarum (Hemiptera: Pentatomidae) for Parasitoid of Eurygaster integriceps by Use of Parasitization by Trissolcus nigripedius Artificial Diet. Pak. J. Biol. Sci., 9: 2040-2047. (Hymenoptera: Scelionidae). Biol. Control, 43 : 45. Silva, F. A. C. and Panizzi, A. R. 2008. The 287-293. Adequacy of Artificial Oviposition Substrates 34. Mainali, B. P., Kim, S. and Lim, U. T. 2012. for Laboratory Rearing of Piezodorus guildinii Effects of Combining Releases of Non-viable (Westwood) (Heterpotera, Pentatomidae). Host Eggs with Insecticide Application on Revi. Bras. Entomol ., 52 : 131-134. Riptortus pedestris Population and Its Egg 46. Strand, M. R., Vinson, S. B., Nettles, W. C. Parasitoid. J. Asia-Pacific Entomol., 15 : 299- and Xie, Z. N. 1988. In vitro Culture of the 305. Egg Parasitoid Telenomus heliothidis : The 35. Massoni, F. and Frana, J. 2006. Enemigos Role of Teratocytes and Medium Consumption Naturales del Complejo de Chinches Fitófagas in Development. Entomol. Exp. Appl., 46 : 71- y Evaluación de su Acción Ecológica en un 78. Cultivo de Soja. Publicación Miscelánea INTA 47. Sujii, E. R., Pires, C. S., Schimidt, F. V. G., EE Rafaela, 106 : 163-165. Armando, M. S., Moreira Borges, M. M., 36. Molinari, A. 2005. Control Biológico. Guimarães Carneiro, R. and Viana Valle, J. C. Especies Entomófagas en Cultivos Agrícolas . 2002. Controle Biológico de Insetos-praga na Ediciones INTA, Centro Regional Santa Fe. Soja Orgânica do Distrito Federal. Cadernos EEA Oliveros, Santa Fe, Argentina, 80 PP. de Ciência Tecnologia, 19 : 299-312. 37. Noble, N. S. 1937. An Egg Parasite of the 48. Temple, J. H., Davis, J. A., Hardke, J. T., Green Vegetable Bug. Agric. Gazette New Moore, J. and Leonard, B. R. 2013a. South Wales, 48 : 337-341. Susceptibility of Southern Green Stink Bug 38. Nordlund, D. A., Lewis, W. J. and Gueldner, and Redbanded Stink Bug to Insecticides in R. C. 1983. Kairomones and Their Use for Soybean Field Experiments and Laboratory Management of Entomophagous Insects. XIV. Bioassays. Southwest. Entomol., 38 : 393-406. Response of Telenomus remus to Abdominal 49. Temple, J. H., Davis, J. A., Micinski, S., Tips of Spodoptera frugiperda , (Z)-9- Hardke, J. T., Price, P. and Leonard, B. R. tetradecene-1-ol Acetate and (Z)-9-dodecene- 2013b. Species Composition and Seasonal 1-ol Acetate. J. Chem. Ecol., 9: 695-701. Abundance of Stink Bugs (Hemiptera: 39. Orr, D. B. 1988. Scelionid Wasps as Pentatomidae) in Louisiana Soybean. Environ. Biological Control Agents: A Review. Fla. Entomol., 42 : 648-657. Entomol., 71 : 506-528. 50. van Lenteren, J. C. and Bueno, V. H. P. 2003. 40. Panizzi, A. R., Postali, J. R., Santos, C. H. and Augmentative Biological Control of Rodriguez Carvalho, D. 2000. Rearing the in Latin America. BioControl. , 48 : Southern Green Stink Bug Using an Artificial 123-139. Dry Diet and an Artificial Plant. Pesqui. 51. Volkoff, N., Vinson, S. B., Wu, Z. X. and Agropecu. Bras., 35 : 1709-1715. Nettles, W. C. J. 1992. In vitro Rearing of 41. Panizzi, A. R. and Slansky, F. 1985. Review of Trissolcus basalis [Hym., Scelionidae] an Egg Phytophagous pentatomids (Hemiptera: Parasitoid of Nezara viridula [Hem., Pentatomidae) Associated with Soybean in the Pentatomidae]. Entomophaga , 37 : 141-148. Americas. Fla. Entomol., 68 : 184-214. 52. Zar, J. H. 1996. Biostatistical Analysis. 3rd 42. Parra, J. R. P. and Cônsoli, F. L. 2009. Criação Edition, Prentice Hall, New Jersey, USA, 662 Massal e Controle de Qualidade de PP. Parasitoides de Ovos. In: “ Controle Biológico 53. Zerbino, M. S., Altier, N. A. and Panizzi, A. R. de Pragas: Produção Massal e Controle de 2013. Effect of Photoperiod and Temperature Qualidade ”; (Ed.): Bueno, V. H. P.. Editora on Nymphal Development and Adult UFLA, Lavras, Brazil, PP. 169-198. Reproduction of Piezodorus guildinii 43. Serra, G. V. and La Porta, N. C. 2001. (Heteroptera: Pentatomidae). Fla. Entomol., Aspectos Biológicos y Reproductivos de 96 : 572-582. Piezodorus guildinii (West.) (Hemiptera: Pentatomidae) en Condiciones de Laboratorio. Agriscientia, 18 : 51-57.

1515 ______Fernanda Cingolani et al.

Piezodorus guildinii ( Hemiptera: Pentatomidae ) Telenomus podisi (Hymenoptera: Platygastridae)

. . !" . .# $% & ' .

()*

Piezodorus guildinii (Hemiptera: Pentatomidae ) Telenomus podisi (Hymenoptera: Platygastridae ) ! " # $ %! &'( ) # * +, - # !./ 0%1 2 3%.1 . - 3.% (, #(5 ! %! %$( * +, !-0 2 . ! 6+, +, - % 7.81 2 9 :%; 2 . 0' <0 %! ( % T. podisi F1 . =. &'( !"> ? .% @, . #(5! %! %$( P. guildini i 0 ( #J%! 18- D %! EF =, ? 24 # * +,:%A 2 # .! MN . %$( ( N=29 ) # . ( N=28 ) 2 : ( N=53 !, ) " K # !F % , %! . !! % , %! T. podisi ! # ? 48 %! * +, 40 !. %! ! F 0( #J%! 24 K% %! . EF =, ? 24 # * +, # +, !% %! 0 :P0( Q . !-0 ( N=55 ) (! %$( !% %! (R0() ( %S . =. &'( ( parasitism ) >$( %0% (! #(5! %! 0, '( :F !*+ %.! 7* .! !*( T +, 0 %! % # ! T +, ( >$( %0% '( : ! R0( ( preimaginal ) U> &; % %.! . ( @, /, F2 %! U> &; % %.! < . :( K '( :R0( %S *( . F1 V= K # P. guildinii ! T +, .% T. podisi !"> ? ) .$( %; F!*+ %.! : .! , #(5! %! * +, %$( :! !*( T +, 0 %! % 3.% 2 :%; 2 .5 %.X % R0( %S Y W( *( . % 40 , >$( W( # 0! #?%* %! >$( %0% , . * (0' [5 %.! %! 8. # ( 3%.1 . - ( # (, * % # ( %!

1516