Section 1 Biological Control

**3**

**Chapter 1**

**Abstract**

*variator*.

**1. Introduction**

viable method of crop protection.

**2. Hymenoptera parasitoid**

Lepidopter Parasitoidea

*Hassan-Ali Vahedi, Jabbar Valipour and Abbas Ali Zamani*

Parasitoids have a long history of pest management, specially for control of economical important lepidopteran pests, such as Noctuidae, Tortricidae, and Pyralidae. The two major parasitoids super families Trichogrammatidae and Braconidae in relation to biocontrol of lepidopteron pests are important. In this chapter: (i) the mass production of the moth egg parasitoid, *Trichogramma brassicae* Bezdenko, 1968 (Hymenoptera, Trichogrammatidae), which has been known to occur in Iran and attack many important hosts Lepidoptera, such as the chickpea pod borer, *Helicoverpa armigera* (Hübner, 1808), Carob moth, *Ectomyelois ceratoniae* (Zeller, 1839), and the codling moth, *Cydia pomonella* (Linnaeus, 1758) in the region and (ii) biology and parasitism behavior of *Bracon* (*Glabrobracon*) *variator* Nees, 1811, as larval ectoparasitoid of *Cydia johanssoni* Aarvik and Karsholt, 1993 (Lepidoptera: Tortricidae), a seed feeder of *Anagyris foetida* Linnaeus, 1758 (Fabaceae) at forest habitats in Western Iran, are discussed. Host development was arrested immediately upon parasitism. The dissected capsules show each adult female lay three eggs close to host larvae inside seed case. Video films and photographs of the behavior as research documents were recorded. Both aestivation and hibernation of the parasitoid occur in the parasitization rate on *C. johanssoni* averaged 18.77% ± 3.80, during second generation of the parasitoid wasp, *B.* (*G.*)

**Keywords:** biological agents, Chalcidoidea, ectoparasitoid, Ichneumonoidea, Iran

Many Lepidoptera larvae are economically important to agriculture and forestry ecosystems. For example, Noctuidae, Pyralidae and Tortricidae. Many Hymenoptera parasitoids, such as Trichogrammatidae and Braconidae are an important potential bio-control agent for control of moth pests. Recently, attention has been focused on this strategy due to increased consumer concern with pesticide residues in food products and a wide-ranging negative impact of chemical insecticides to the environment. Thus, using parasitoid Hymenoptera can be a safe and

Hymenoptera is the third largest and perhaps the most beneficial to humans of all insect orders. It has around 320,000 species, mostly more than 75% (240,000) are parasitoids (in 12 super families). Parasitoid wasps are highly diverse and specialized to attack a particular host life stage (egg, larvae, pupae and adults) of

## **Chapter 1**

## Lepidopter Parasitoidea

*Hassan-Ali Vahedi, Jabbar Valipour and Abbas Ali Zamani*

## **Abstract**

Parasitoids have a long history of pest management, specially for control of economical important lepidopteran pests, such as Noctuidae, Tortricidae, and Pyralidae. The two major parasitoids super families Trichogrammatidae and Braconidae in relation to biocontrol of lepidopteron pests are important. In this chapter: (i) the mass production of the moth egg parasitoid, *Trichogramma brassicae* Bezdenko, 1968 (Hymenoptera, Trichogrammatidae), which has been known to occur in Iran and attack many important hosts Lepidoptera, such as the chickpea pod borer, *Helicoverpa armigera* (Hübner, 1808), Carob moth, *Ectomyelois ceratoniae* (Zeller, 1839), and the codling moth, *Cydia pomonella* (Linnaeus, 1758) in the region and (ii) biology and parasitism behavior of *Bracon* (*Glabrobracon*) *variator* Nees, 1811, as larval ectoparasitoid of *Cydia johanssoni* Aarvik and Karsholt, 1993 (Lepidoptera: Tortricidae), a seed feeder of *Anagyris foetida* Linnaeus, 1758 (Fabaceae) at forest habitats in Western Iran, are discussed. Host development was arrested immediately upon parasitism. The dissected capsules show each adult female lay three eggs close to host larvae inside seed case. Video films and photographs of the behavior as research documents were recorded. Both aestivation and hibernation of the parasitoid occur in the parasitization rate on *C. johanssoni* averaged 18.77% ± 3.80, during second generation of the parasitoid wasp, *B.* (*G.*) *variator*.

**Keywords:** biological agents, Chalcidoidea, ectoparasitoid, Ichneumonoidea, Iran

## **1. Introduction**

Many Lepidoptera larvae are economically important to agriculture and forestry ecosystems. For example, Noctuidae, Pyralidae and Tortricidae. Many Hymenoptera parasitoids, such as Trichogrammatidae and Braconidae are an important potential bio-control agent for control of moth pests. Recently, attention has been focused on this strategy due to increased consumer concern with pesticide residues in food products and a wide-ranging negative impact of chemical insecticides to the environment. Thus, using parasitoid Hymenoptera can be a safe and viable method of crop protection.

## **2. Hymenoptera parasitoid**

Hymenoptera is the third largest and perhaps the most beneficial to humans of all insect orders. It has around 320,000 species, mostly more than 75% (240,000) are parasitoids (in 12 super families). Parasitoid wasps are highly diverse and specialized to attack a particular host life stage (egg, larvae, pupae and adults) of

most arthropods, mainly insects. Major species richness of parasitoid wasps is in Ichneumonoidea 100,000 (which include Braconidae 40,000 and Ichneumonidae 60,000 known species) and Chalcidoidea 22,000 known species; Most species of Chalcidoidea are tiny, <3 mm in length; as a result, they can be difficult to collect and study. Detailed study an estimated more than 500,000 species in existence belong to the Chalcidoidea; within this superfamily, Trichogrammatidae are the smallest, ~0.2 mm in length, which includes 83 genera and 839 known species. Some *Trichogramma* species and strains have a wide host range of insect eggs, while others have strong preference for the eggs of a particular moth species and many successful biological control programs have involved the introduction of highly specific parasitoids [1–4]. The two major parasitoids groups, Trichogrammatidae and Braconidae in relation to biocontrol of lepidopteron pests are important. This chapter is concerned mass production of the egg parasitoid, *T. brassicae* Bezdenko, 1968 (Trichogrammatidae); addition refer to biology and parasitism behavior of *B.* (*G.*) *variator* Nees, 1812 (Braconidae), in relation to lepidopteran pests.

## **3. Trichogrammatidae**

Most prominent species of *Trichogramma* are mostly amenable for insectarium mass production (**Figures 1, 2** and **6**) on factitious hosts like the grain moth, *Sitotroga cerealella* (Olivier, 1789), which it selves is mass produced on factory scales and is being used for biological control of noxious Lepidopterous pests of crops worldwide (**Figures 1, 2** and **6**). *Trichogramma* adults are typically free-living and the females are responsible for finding host insects for their progeny.

## **3.1** *Trichogramma* **biology**

The development of all *Trichogramma* spp. is very similar. The eggs hatch in about 24 h and the parasite larvae develop through three instars. These are followed by a prepupa, when the adult characters form, and a pupa. At the beginning of the third larval instars, the host egg turns black due to the deposition of black granules at the inner surface of the chorion, an invaluable diagnostic character for parasitized eggs. After about 5 days, the adult wasps emerge from the pupae and escape the bollworm egg by chewing a circular hole in the egg shell (**Figure 2**). *Trichogramma* overwinter as immature forms in host eggs. This short life cycle allows multi generations per year, and rapid population increase [5]. Hence, early season releases

#### **Figure 1.**

*General view drawing of the moth egg parasitoid, Trichogramma australicum Girault, 1912 (Hymenoptera: Trichogrammatidae), adult male ×120 [6].*

**5**

**Figure 4.**

*Lepidopter Parasitoidea*

**Figure 2.**

**Figure 3.**

*diameter 0.4–0.6 mm). Original.*

*and 0.27 ± 0.08 cm wide). Original.*

*DOI: http://dx.doi.org/10.5772/intechopen.86381*

*(Hübner, 1808) [5] (moth egg diameter 0.4–0.6 mm).*

produce large populations positioned to fight pest invasions. Unparasitized eggs remain light until black at hatching stage (**Figure 3**). For example, the development time for *T. australicum* Girault, 1912 (Hymenoptera: Trichogrammatidae) on the rice moth, *Corcyra cephalonica* (Stainton, 1866) (Lepidoptera: Galleriidae) at 28 ± 2 from egg to adult are completed in 14 days [5, 6]. The early stages of developed eggs (**Figure 3B**) are more suitable for parasite development. Older bollworm eggs, especially those in which the head capsule of the larva is visible, are not usually parasitized and if they are, parasite survival is much lower (**Figure 3C**) [7].

*Life cycle of the moth egg parasitoid, Trichogramma developing on the bollworm egg (Helicoverpa armigera)* 

*Light micrographs of development of the pod borer eggs (unparasitized), Helicoverpa armigera (Hübner, 1808) showing the embryonic developmental sequences: (A) newly laid egg (yellowish-white); (B) 2 days old egg (tan egg); (C) egg before hatching or black head stage (larva visible) and (D) hatching stage (larva) (moth egg* 

*Images general view of the chickpea pod borer, Helicoverpa armigera (Hübner, 1808) (Lepidoptera: Noctuidae): (Right) 5th larval stage off the chickpea and (Left) The adult stage (Lepidoptera: Noctuidae); (adult wingspan 3.5–4 cm; 1.5–1.9 cm long and the mature larva or last instars was about 2.8 ± 0.05 mm long* 

## *Lepidopter Parasitoidea DOI: http://dx.doi.org/10.5772/intechopen.86381*

produce large populations positioned to fight pest invasions. Unparasitized eggs remain light until black at hatching stage (**Figure 3**). For example, the development time for *T. australicum* Girault, 1912 (Hymenoptera: Trichogrammatidae) on the rice moth, *Corcyra cephalonica* (Stainton, 1866) (Lepidoptera: Galleriidae) at 28 ± 2 from egg to adult are completed in 14 days [5, 6]. The early stages of developed eggs (**Figure 3B**) are more suitable for parasite development. Older bollworm eggs, especially those in which the head capsule of the larva is visible, are not usually parasitized and if they are, parasite survival is much lower (**Figure 3C**) [7].

#### **Figure 2.**

*Trends in Integrated Insect Pest Management*

**3. Trichogrammatidae**

**3.1** *Trichogramma* **biology**

most arthropods, mainly insects. Major species richness of parasitoid wasps is in Ichneumonoidea 100,000 (which include Braconidae 40,000 and Ichneumonidae 60,000 known species) and Chalcidoidea 22,000 known species; Most species of Chalcidoidea are tiny, <3 mm in length; as a result, they can be difficult to collect and study. Detailed study an estimated more than 500,000 species in existence belong to the Chalcidoidea; within this superfamily, Trichogrammatidae are the smallest, ~0.2 mm in length, which includes 83 genera and 839 known species. Some *Trichogramma* species and strains have a wide host range of insect eggs, while others have strong preference for the eggs of a particular moth species and many successful biological control programs have involved the introduction of highly specific parasitoids [1–4]. The two major parasitoids groups, Trichogrammatidae and Braconidae in relation to biocontrol of lepidopteron pests are important. This chapter is concerned mass production of the egg parasitoid, *T. brassicae* Bezdenko, 1968 (Trichogrammatidae); addition refer to biology and parasitism behavior of *B.*

(*G.*) *variator* Nees, 1812 (Braconidae), in relation to lepidopteran pests.

Most prominent species of *Trichogramma* are mostly amenable for insectarium

The development of all *Trichogramma* spp. is very similar. The eggs hatch in about 24 h and the parasite larvae develop through three instars. These are followed by a prepupa, when the adult characters form, and a pupa. At the beginning of the third larval instars, the host egg turns black due to the deposition of black granules at the inner surface of the chorion, an invaluable diagnostic character for parasitized eggs. After about 5 days, the adult wasps emerge from the pupae and escape the bollworm egg by chewing a circular hole in the egg shell (**Figure 2**). *Trichogramma* overwinter as immature forms in host eggs. This short life cycle allows multi

generations per year, and rapid population increase [5]. Hence, early season releases

*General view drawing of the moth egg parasitoid, Trichogramma australicum Girault, 1912 (Hymenoptera:* 

mass production (**Figures 1, 2** and **6**) on factitious hosts like the grain moth, *Sitotroga cerealella* (Olivier, 1789), which it selves is mass produced on factory scales and is being used for biological control of noxious Lepidopterous pests of crops worldwide (**Figures 1, 2** and **6**). *Trichogramma* adults are typically free-living and

the females are responsible for finding host insects for their progeny.

**4**

**Figure 1.**

*Trichogrammatidae), adult male ×120 [6].*

*Life cycle of the moth egg parasitoid, Trichogramma developing on the bollworm egg (Helicoverpa armigera) (Hübner, 1808) [5] (moth egg diameter 0.4–0.6 mm).*

#### **Figure 3.**

*Light micrographs of development of the pod borer eggs (unparasitized), Helicoverpa armigera (Hübner, 1808) showing the embryonic developmental sequences: (A) newly laid egg (yellowish-white); (B) 2 days old egg (tan egg); (C) egg before hatching or black head stage (larva visible) and (D) hatching stage (larva) (moth egg diameter 0.4–0.6 mm). Original.*

#### **Figure 4.**

*Images general view of the chickpea pod borer, Helicoverpa armigera (Hübner, 1808) (Lepidoptera: Noctuidae): (Right) 5th larval stage off the chickpea and (Left) The adult stage (Lepidoptera: Noctuidae); (adult wingspan 3.5–4 cm; 1.5–1.9 cm long and the mature larva or last instars was about 2.8 ± 0.05 mm long and 0.27 ± 0.08 cm wide). Original.*

#### **Figure 5.**

*Images general view of the pomegranate calyx worm, Ectomyelois ceratoniae (Zeller, 1839) (Lepidoptera: Pyralidae) and infested pomegranate fruit: Right to left. The adult stage of E. ceratoniae; 5th larval stage of E. ceratoniae off the pomegranate; infested split pomegranate fruit and applied cardboard capsule for infested orchards; (adult wingspan 1.6–2.5 cm, 1.5–1.8 cm long and the mature larva or last instars was about 1.8 ± 0.07 cm long and 0.75 ± 0.05 cm wide). Original.*

Recently, mass rearing techniques have been developed for several parasitoids of insects pests, including, *Trichogramma* (Chalcidoidea: Trichogrammatidae). Currently, several laboratories are actively engaged in producing *Trichogramma* on local scale. These biological agents, parasitoids, are mass produced and released for controlling variety of insect pests including, the chickpea pod borer, *H. armigera* (Hübner, 1808) and the Pomegranate calyx worm, *E. ceratoniae* (Zeller, 1839), which maintain populations below economic levels [8, 9]. The chickpea pod borer, *H. armigera* (Hübner, 1808), (**Figure 4**) produces 52.5% losses [10] and more than 60% losses in the chickpea grower area every year [11]. It attacks on other Legume variety of economically important crops.

The Carob moth, *E. ceratoniae*, which known "Pomegranate calyx worm" (**Figure 5**). It is a pest that attack on variety of fruit crops such as Pomegranate, *Punica granatum* Linnaeus, 1880; Apples*, Malus domestica* Borkh, 1803; date palm, *Phoenix dactylifera* Linnaeus, 1753; Cultivated pistachio*, Pistacia vera* Linnaeus, 1753 [12, 13]. Split pomegranate fruit are more prone to infestation by larva as penetration into the fruit occurs more readily. The adult female deposits its eggs in already split fruits or on the skin or calyx of pomegranate fruit. It is widely distributed and occurs in Iran and close countries [12].

## **4.** *Trichogramma brassicae* **Bezdenko, 1968 (Hymenoptera: Trichogrammatidae)**

The moth egg parasitoid, *T. brassicae* Bezdenko, 1968 is reared in private or government owned insectaries and released annually in the agricultural crops and orchards in Iran [14]. For example, the development time for *T. brassicae* Bezdenko, 1968, on the grain moth, *Sitotroga cerealella* Olivier, 1789 (Lepidoptera: Galleriidae) at 28 ± 2°C from egg to adult is completed in 14 days [14].

*T. brassicae* Bezdenko, 1968, pupae can be programmed to enter an overwintering condition of arrested development called diapauses. Once in diapauses, wasp pupae can be stored for up to 9 months so that the large demand for *Trichogramma* during the summer can be met [14, 15].

Cardboard capsules containing host eggs with developing *Trichogramma* are applied to release of *Trichogramma* in the chickpea farms or pomegranate orchards (**Figure 5**).

Released *Trichogramma* are at different developmental stages so that adults emerge from the capsules over several days. This increases the time interval between applications.

Two releases each at a rate of 460,000 pupae per hectare are made beginning at the first moth flight as determined by light traps. Chickpea pod borer, eggs hatch

**7**

*Lepidopter Parasitoidea*

untreated fields [15].

field releases.

mate.

Bezdenko, 1968 (**Figure 6**).

on the each egg sheet.

UV tubes) for 45 min at a distance of 5 cm.

*DOI: http://dx.doi.org/10.5772/intechopen.86381*

the Carob moth, *E. ceratoniae* (Zeller, 1839).

**5. Parasitism behavior of** *Trichogramma*

**6. Materials and methods: Mass production technique**

after about 5–6 days and the egg-laying period continues for 3–6 weeks. In-field reproduction of released parasites is believed to be important in providing residual control of eggs deposited after the second release. Field evaluations in Germany have shown releases result in a 70–93% reduction in corn borer larvae relative to

In western parts of Iran, releases of *Trichogramma* are a parts of integrated pest management in controlling the chickpea pod borer, *H. armigera* (Hübner, 1808) and

Parasitoid attributes include: The ability to parasitize and develop in the target host egg, the species' preference for the target host egg, total egg mortality caused by parasitism, adult feeding, fecundity, development rate, sex ratio, and longevity releases [16, 17]. These characters are important in mass-rearing programs and then

*Trichogramma* spp. are most famous biocontrol agents and widely distributed in the world. *Trichogramma* drills a hole through the egg-shell and inserts two to three eggs into eggs of 200 pest moth species, including *Helicoverpa* spp., *Chilo* spp. the pink bollworm (*Pectinophora gossypiella* Saunders, 1844 and etc.) and preventing neonate larvae from hatching out and devouring crops. These parasitoids wasps are so small, <1 mm long; moth egg size, and hence how many of their own eggs to lay, is calculated by timing walks across moth egg surfaces. *Trichogramma* larvae eat out the insides of pest eggs, pupate, and cut an exit hole in moth eggshells for winged adults to squeeze through. Males emerge first, wait for females, and immediately

Native parasitoid has proved to be one of the most potent egg parasitoid for so many important caterpillar pests. It is reared on factitious hosts such as, the flour moth, *Ephestia kuehniella* Zeller, 1879 (Lep.: Pyralidae), the grain moth, *S. cerealella* (Olivier, 1789), (Lep.: Gelechiidae) and other stored grain pests. The eggs of above hosts are utilized in mass rearing of this parasitoid [18]. *Sitotroga* eggs, test tubes, egg cards, refrigerator, UV chamber (fluorescent tube light, 15 W/Luminal flow), honey solution 20%, gam arabic, camel hair brush, Glass and adults of the grain moth, *S. cerealella* (Olivier, 1789), (Lep.: Gelechiidae). Following rearing procedure are involved in mass production of *T. brassicae*

Providing the Grain moth eggs: (i) For prevention from hatching of the eggs during and after parasitization of eggs, they should be exposed to UV rays (15 watt

Processed egg sheet is than placed the plastic box. Introduce 6 days old duly parasitized egg cards to adult females of *Trichogramma* for 24 h. The parasitized and unparasitized eggs in each of box/containers should be in the ratio of 1:6 (parasitoid:host) to have optimum parasitization. Close the container properly for preventing escape of parasitoids. The parasitoids emerged in the container will parasitize the unparasitized eggs of the grain moth, *S. cerealella* (Olivier, 1789)*.*

Parasitoid species name, date of release of parasitoid for parasitization, expected date of emergence and Institute name and name of technical person should be given

#### *Lepidopter Parasitoidea DOI: http://dx.doi.org/10.5772/intechopen.86381*

*Trends in Integrated Insect Pest Management*

**Figure 5.**

variety of economically important crops.

*1.8 ± 0.07 cm long and 0.75 ± 0.05 cm wide). Original.*

occurs in Iran and close countries [12].

during the summer can be met [14, 15].

**Trichogrammatidae)**

Recently, mass rearing techniques have been developed for several parasitoids of insects pests, including, *Trichogramma* (Chalcidoidea: Trichogrammatidae). Currently, several laboratories are actively engaged in producing *Trichogramma* on local scale. These biological agents, parasitoids, are mass produced and released for controlling variety of insect pests including, the chickpea pod borer, *H. armigera* (Hübner, 1808) and the Pomegranate calyx worm, *E. ceratoniae* (Zeller, 1839), which maintain populations below economic levels [8, 9]. The chickpea pod borer, *H. armigera* (Hübner, 1808), (**Figure 4**) produces 52.5% losses [10] and more than 60% losses in the chickpea grower area every year [11]. It attacks on other Legume

*Images general view of the pomegranate calyx worm, Ectomyelois ceratoniae (Zeller, 1839) (Lepidoptera: Pyralidae) and infested pomegranate fruit: Right to left. The adult stage of E. ceratoniae; 5th larval stage of E. ceratoniae off the pomegranate; infested split pomegranate fruit and applied cardboard capsule for infested orchards; (adult wingspan 1.6–2.5 cm, 1.5–1.8 cm long and the mature larva or last instars was about* 

The Carob moth, *E. ceratoniae*, which known "Pomegranate calyx worm" (**Figure 5**). It is a pest that attack on variety of fruit crops such as Pomegranate, *Punica granatum* Linnaeus, 1880; Apples*, Malus domestica* Borkh, 1803; date palm, *Phoenix dactylifera* Linnaeus, 1753; Cultivated pistachio*, Pistacia vera* Linnaeus, 1753 [12, 13]. Split pomegranate fruit are more prone to infestation by larva as penetration into the fruit occurs more readily. The adult female deposits its eggs in already split fruits or on the skin or calyx of pomegranate fruit. It is widely distributed and

The moth egg parasitoid, *T. brassicae* Bezdenko, 1968 is reared in private or government owned insectaries and released annually in the agricultural crops and orchards in Iran [14]. For example, the development time for *T. brassicae* Bezdenko, 1968, on the grain moth, *Sitotroga cerealella* Olivier, 1789 (Lepidoptera: Galleriidae)

*T. brassicae* Bezdenko, 1968, pupae can be programmed to enter an overwintering condition of arrested development called diapauses. Once in diapauses, wasp pupae can be stored for up to 9 months so that the large demand for *Trichogramma*

Cardboard capsules containing host eggs with developing *Trichogramma* are applied to release of *Trichogramma* in the chickpea farms or pomegranate orchards

Released *Trichogramma* are at different developmental stages so that adults emerge from the capsules over several days. This increases the time interval

Two releases each at a rate of 460,000 pupae per hectare are made beginning at the first moth flight as determined by light traps. Chickpea pod borer, eggs hatch

**4.** *Trichogramma brassicae* **Bezdenko, 1968 (Hymenoptera:** 

at 28 ± 2°C from egg to adult is completed in 14 days [14].

**6**

(**Figure 5**).

between applications.

after about 5–6 days and the egg-laying period continues for 3–6 weeks. In-field reproduction of released parasites is believed to be important in providing residual control of eggs deposited after the second release. Field evaluations in Germany have shown releases result in a 70–93% reduction in corn borer larvae relative to untreated fields [15].

In western parts of Iran, releases of *Trichogramma* are a parts of integrated pest management in controlling the chickpea pod borer, *H. armigera* (Hübner, 1808) and the Carob moth, *E. ceratoniae* (Zeller, 1839).

Parasitoid attributes include: The ability to parasitize and develop in the target host egg, the species' preference for the target host egg, total egg mortality caused by parasitism, adult feeding, fecundity, development rate, sex ratio, and longevity releases [16, 17]. These characters are important in mass-rearing programs and then field releases.

## **5. Parasitism behavior of** *Trichogramma*

*Trichogramma* spp. are most famous biocontrol agents and widely distributed in the world. *Trichogramma* drills a hole through the egg-shell and inserts two to three eggs into eggs of 200 pest moth species, including *Helicoverpa* spp., *Chilo* spp. the pink bollworm (*Pectinophora gossypiella* Saunders, 1844 and etc.) and preventing neonate larvae from hatching out and devouring crops. These parasitoids wasps are so small, <1 mm long; moth egg size, and hence how many of their own eggs to lay, is calculated by timing walks across moth egg surfaces. *Trichogramma* larvae eat out the insides of pest eggs, pupate, and cut an exit hole in moth eggshells for winged adults to squeeze through. Males emerge first, wait for females, and immediately mate.

## **6. Materials and methods: Mass production technique**

Native parasitoid has proved to be one of the most potent egg parasitoid for so many important caterpillar pests. It is reared on factitious hosts such as, the flour moth, *Ephestia kuehniella* Zeller, 1879 (Lep.: Pyralidae), the grain moth, *S. cerealella* (Olivier, 1789), (Lep.: Gelechiidae) and other stored grain pests. The eggs of above hosts are utilized in mass rearing of this parasitoid [18]. *Sitotroga* eggs, test tubes, egg cards, refrigerator, UV chamber (fluorescent tube light, 15 W/Luminal flow), honey solution 20%, gam arabic, camel hair brush, Glass and adults of the grain moth, *S. cerealella* (Olivier, 1789), (Lep.: Gelechiidae). Following rearing procedure are involved in mass production of *T. brassicae* Bezdenko, 1968 (**Figure 6**).

Providing the Grain moth eggs: (i) For prevention from hatching of the eggs during and after parasitization of eggs, they should be exposed to UV rays (15 watt UV tubes) for 45 min at a distance of 5 cm.

Parasitoid species name, date of release of parasitoid for parasitization, expected date of emergence and Institute name and name of technical person should be given on the each egg sheet.

Processed egg sheet is than placed the plastic box. Introduce 6 days old duly parasitized egg cards to adult females of *Trichogramma* for 24 h. The parasitized and unparasitized eggs in each of box/containers should be in the ratio of 1:6 (parasitoid:host) to have optimum parasitization. Close the container properly for preventing escape of parasitoids. The parasitoids emerged in the container will parasitize the unparasitized eggs of the grain moth, *S. cerealella* (Olivier, 1789)*.*

#### **Figure 6.**

*Mass rearing procedure schematic: conventional production of egg parasitoid, Trichogramma brassicae, and its laboratory host, the eggs of grain moth, Sitotroga cerealella (Olivier, 1789)*.

Approaching the time of emergence of parasitoid, the egg color becomes blackish. Mating of sexes is essential for obtaining sex ratio, male:female, 1:1. Thus, mating is occurred in container.

Unparasitized eggs get hatched, into larvae (of host), such larvae should be removed from the container with the help of camel hair brush. The host larvae may destroy the eggs hence they should be removed from the container. The parasitoid complete its life cycle with 7–8 days.

**9**

*Lepidopter Parasitoidea*

*DOI: http://dx.doi.org/10.5772/intechopen.86381*

**(Hymenoptera: Braconidae)**

the outside of the caterpillars.

video films and photographs of the wasp behavior.

microscope to observe the number of parasitoid eggs laid.

[25]. Braconids also vary greatly in their biology [26–28].

tion observed.

**8. Braconidae**

After 4 days of parasitization, parasitized eggs of the grain moth, *S. cerealella* (Olivier, 1789), start changing their color from creamy white to blackish due to accumulation of urate granules. Such egg sheets are removed and stored in refrigerator at 10°C temperature for about 20–30 days for our convenience, duskiness

**oviposition behavior of** *Bracon* **(***Glabrobracon***)** *variator* **Nees, 1812,** 

In order to determine parasitism rate of the ectoparasitoid, *B.* (*G.*) *variator* Nees, during first third of June, at least 100 infested seed pods (the stinking bean trefoil, *Anagyris foetida* Linnaeus, 1758 (Fabaceae)) by larva of *Cydia johanssoni* (Lepidoptera: Tortricidae) were collected daily and the number of parasitized larvae was counted and parasitism rate was calculated. The parasitoid was visible on

In order to observe parasitism behavior, several live female wasps were collected along with pods containing unparasitized larvae and transported to laboratory. One adult female wasp was released into a petri dish with an infested pod and oviposi-

A digital camera, model Canon, PowerShot S3 IS and a digital Sony EXwave HAD camera connected to a stereomicroscope, model Blue Light, were used to take

Immediately after oviposition, the seed capsules were dissected, under a stereo-

The Braconidae constitute one of the most species-rich hymenopterous families,

Taxonomic significance of the genus *Bracon* Fabricius, 1804: The width of the hypoclypeal depression may be important at subgenera level; presence or absence of occipital carina and shape of first metasomal tergite (t1) (flattened or not flattened lateral parts) [21]. *Bracon* genus is measured as a paraphyletic or also a polyphyletic group, mainly of small and middle-sized species [22] divided over several subgenera [23, 24]. For example, *Bracon* fauna of Iran include five subgenera: *Bracon* (*Bracon*) kozak Telenga, 1936; *B.* (*Glabrobracon*) Fahringer 1927; *B.* (*Habrobracon*) Ashmead, 1895; *B.* (*Asiabracon*) Tobias 1957 and *B.* (*Orthobracon*) Fahringer, 1927

Different species of parasitoids attack different life stages of the pest. For example,

*B.* (*G.*) *variator* Nees, 1812, are larval ectoparasitoid and prefers 3rd–5th instars larvae and each adult female laid three eggs close to host larvae (**Figure 7**), inside seed case [32]. Most parasitoids behaviors are so identical and specialized that they can attack only a particular host life stage i.e., they attack eggs (eggs parasitoids), larvae (larval parasitoids), Pupae (pupal parasitoids), or adult (adult's parasitoids) [1]. There are considerable variations in insect parasitoids parasitism. These may be

with 48 subfamilies, more than 1050 genera and about 17,600 species described worldwide. The family has two major lineages: (a) the cyclostome braconids, most of which are idiobiont ectoparasitoids of concealed Lepidoptera and Coleoptera larvae although many are koinobiont endoparasitoids of Diptera and Hemiptera and (b) the noncyclostome braconids which are all endoparasitoids and are generally

koinobionts, typically attacking the early instars of their hosts [19, 20].

point of view. The egg sheets thus preserved can be marked to farmers.

**7. Method of calculation of parasitism rate and observation of** 

#### *Lepidopter Parasitoidea DOI: http://dx.doi.org/10.5772/intechopen.86381*

*Trends in Integrated Insect Pest Management*

**8**

**Figure 6.**

occurred in container.

complete its life cycle with 7–8 days.

Approaching the time of emergence of parasitoid, the egg color becomes blackish. Mating of sexes is essential for obtaining sex ratio, male:female, 1:1. Thus, mating is

*Mass rearing procedure schematic: conventional production of egg parasitoid, Trichogramma brassicae, and its* 

*laboratory host, the eggs of grain moth, Sitotroga cerealella (Olivier, 1789)*.

Unparasitized eggs get hatched, into larvae (of host), such larvae should be removed from the container with the help of camel hair brush. The host larvae may destroy the eggs hence they should be removed from the container. The parasitoid

After 4 days of parasitization, parasitized eggs of the grain moth, *S. cerealella* (Olivier, 1789), start changing their color from creamy white to blackish due to accumulation of urate granules. Such egg sheets are removed and stored in refrigerator at 10°C temperature for about 20–30 days for our convenience, duskiness point of view. The egg sheets thus preserved can be marked to farmers.

## **7. Method of calculation of parasitism rate and observation of oviposition behavior of** *Bracon* **(***Glabrobracon***)** *variator* **Nees, 1812, (Hymenoptera: Braconidae)**

In order to determine parasitism rate of the ectoparasitoid, *B.* (*G.*) *variator* Nees, during first third of June, at least 100 infested seed pods (the stinking bean trefoil, *Anagyris foetida* Linnaeus, 1758 (Fabaceae)) by larva of *Cydia johanssoni* (Lepidoptera: Tortricidae) were collected daily and the number of parasitized larvae was counted and parasitism rate was calculated. The parasitoid was visible on the outside of the caterpillars.

In order to observe parasitism behavior, several live female wasps were collected along with pods containing unparasitized larvae and transported to laboratory. One adult female wasp was released into a petri dish with an infested pod and oviposition observed.

A digital camera, model Canon, PowerShot S3 IS and a digital Sony EXwave HAD camera connected to a stereomicroscope, model Blue Light, were used to take video films and photographs of the wasp behavior.

Immediately after oviposition, the seed capsules were dissected, under a stereomicroscope to observe the number of parasitoid eggs laid.

## **8. Braconidae**

The Braconidae constitute one of the most species-rich hymenopterous families, with 48 subfamilies, more than 1050 genera and about 17,600 species described worldwide. The family has two major lineages: (a) the cyclostome braconids, most of which are idiobiont ectoparasitoids of concealed Lepidoptera and Coleoptera larvae although many are koinobiont endoparasitoids of Diptera and Hemiptera and (b) the noncyclostome braconids which are all endoparasitoids and are generally koinobionts, typically attacking the early instars of their hosts [19, 20].

Taxonomic significance of the genus *Bracon* Fabricius, 1804: The width of the hypoclypeal depression may be important at subgenera level; presence or absence of occipital carina and shape of first metasomal tergite (t1) (flattened or not flattened lateral parts) [21]. *Bracon* genus is measured as a paraphyletic or also a polyphyletic group, mainly of small and middle-sized species [22] divided over several subgenera [23, 24]. For example, *Bracon* fauna of Iran include five subgenera: *Bracon* (*Bracon*) kozak Telenga, 1936; *B.* (*Glabrobracon*) Fahringer 1927; *B.* (*Habrobracon*) Ashmead, 1895; *B.* (*Asiabracon*) Tobias 1957 and *B.* (*Orthobracon*) Fahringer, 1927 [25]. Braconids also vary greatly in their biology [26–28].

Different species of parasitoids attack different life stages of the pest. For example, *B.* (*G.*) *variator* Nees, 1812, are larval ectoparasitoid and prefers 3rd–5th instars larvae and each adult female laid three eggs close to host larvae (**Figure 7**), inside seed case [32]. Most parasitoids behaviors are so identical and specialized that they can attack only a particular host life stage i.e., they attack eggs (eggs parasitoids), larvae (larval parasitoids), Pupae (pupal parasitoids), or adult (adult's parasitoids) [1]. There are considerable variations in insect parasitoids parasitism. These may be

#### **Figure 7.**

*Image general view of the larval ectoparasitoid wasp, adult female, Bracon (Glabrobracon) variator Nees, 1812 (Hymenoptera: Braconidae). Original.*

#### **Figure 8.**

*Images general view of the larval ectoparasitoid wasp, Bracon (Glabrobracon) variator Nees, 1812, (Hymenoptera: Braconidae) and its host larva, Cydia johanssoni Aarvik & Karsholt, 1993 (Lepidoptera: Tortricidae): (A) female, oviposition on infested pod of stinking bean trefoil by pyralid seed feeder larva; (B) parasitoid's eggs ((B. (G.) variator)) laid close to the head of host larva (C. johanssoni); (C) developed parasitoid larvae inside the seed and (D) parasitoid larvae ((B. (G.) variator)) feeding upon Pyralid seed feeder larva, Cydia spp. [31].*

idiobiont, whose hosts stop development, when they are parasitized. Idiobionts are either ectoparasitoids that kill their hosts or endoparasitoids that attack immobile host stages such as eggs or pupae. Koinobionts, allow the hosts to continue their development until the parasitoid's offspring matures. Most koinobionts are endoparasitoids of larval stages of insects, although a few are ectoparasitic [27]. Some female parasitoids also use the ovipositor to puncture a host and then feed on the body fluids before selecting other hosts for oviposition, thus causing two different types of mortality in the caterpillar pest population. In some cases, for example, *B.* (*G.*) *variator* Nees, 1812, the egg is laid externally on the body of the host and the larvae also feed externally. This parasitoid wasp is a highly polyphagous gregarious ectoparasitoid that attacks the larvae of a wide range of insects, such as Lepidoptera (**Figure 8**). For example, see [28], *B. (G.) variator* Nees, 1812, paralyzes the larvae of *Hadena bicruris* (Hufnagel, 1766) (Lepidoptera: Noctuidae) before depositing on average 3 eggs on 3rd-5th larval instars [29, 30]. Like *C. johanssoni* Aarvik & Karsholt, 1993, *H. bicruris* (Hufnagel, 1766) is a seed specialist, feeding on *Silene latifolia* Poir. 1789 (Caryophyllaceae) seeds. Lepidoptera larvae, attacks the young seeds before they are shed by the parent plant [32–34]. The larvae of the beech moth, *Cydia fagiglandana* (Zeller, 1841), feed inside the nuts of European beech, *Fagus sylvatica* Linnaeus, 1753 (Fabaceae) causing high seed mortality in South Sweden beech forests [35, 36]. Other economically important species are the pea moth, *C. nigricana* (Fabricius, 1794), attacking legume crops; the spruce seed moth, *C. strobilella* Linnaeus, 1758, attacking

**11**

**Table 1.**

**Stages Egg 1st–3rd** 

*M = male and F = female.*

**larval instars**

*Lepidopter Parasitoidea*

*DOI: http://dx.doi.org/10.5772/intechopen.86381*

more than 3 eggs were laid up to a maximum of six.

and had a very low population in the study area (**Figure 8D**).

generation of *B.* (*G.*) *variator* Nees, 1812 (**Tables 1** and **2**) [31].

Siberia, Russia, Crimea, Iran, Turkey and European country [26].

*grain moth eggs, Sitotroga cerealella Olivier, 1789 (Lepidoptera: Galleriidae) at 28 ± 2°C.*

The parasitization rate on *C. johanssoni* averaged 18.77% ± 3.80, during second

General distribution of this wasp includes: China, Central Asia, Mongolia,

**Prepupa Pupa Adult** 

Day/s 1 4 1 2 7 14 1 1: 1

*The life stages longevity, release period and sex ratio of Trichogramma brassicae Bezdenko, 1968, reared on the* 

**longevity**

**Total Release period**

**Sex ratio M/F**

spruce seed, *Picea* spp.; *C. latiferreana* Walsingham, 1879, attacking fruits oak and the hickory shuckworm moth, *C. caryana* Fitch, 1756, an important pest of pecan [37]. First generation adult parasitoids of *B.* (*G.*) *variator* Nees, 1812, appeared in the first ten days of May, with the second generation appearing about a fortnight later, but adult wasps were most abundant, and its percentage parasitism were highest among larvae, in early June (**Table 2**). Adult activity was greatest during the hottest part of the day (12.00–14.00 hours). The parasitoid population was greatest on pods infested with *C. johanssoni* Aarvik and Karsholt, 1993, larvae, early June*.* Oviposition began after a lengthy search period. The female wasp first inspected the drilling into the pod (**Figure 8A**), using her antennae by tapping the pod. She then moved so that her mesothoracic legs straddled the drilling position, lifted her abdomen and inserted her ovipositor almost vertically into the pod (recorded Video film). Almost always each caterpillar had three eggs laid on it (**Figure 8B**); rarely, it was noted that

The oviposition is not an easy task. The tip of the ovipositor almost always gets stuck to tiny irregularities of the pod surface. When she loses her balance, re-starts oviposition from the beginning. Since the exact point of drilling is crucial and must be recalculated for accuracy. The eggs were white and bacilli-form, with a diameter approximately equal to the tip of a lab needle (**Figure 8B**). Eggs were oviposited directly onto the host larvae. The incubation period is 18 hours at 33°C and relative humidity of 14%. Upon hatching, the parasitoid larva penetrated the caterpillar's cuticle and fed on the body hemolymph for 4–5 days. Upon seed maturation, the parasitoid maturated larva secreted a webbed cocoon within the seed capsule and then pupated. Adults of second generation leave the seed pod in last ten days of May via a hole made by adult. Adults lived as free-living adults for up to 3–4 days until they mated and oviposited. When the larvae of *C. johanssoni* were paralyzed, their movement became reduced. As a result of feeding by the parasitoid larvae, the moth larva was weakened and eventually is reduced to the head capsule and body cuticle. The free living adult wasps fed on resin produced when opens the first larval stage of *C. johanssoni* Aarvik and Karsholt, 1993, in the pod. The parasitoid aestivation and hibernation was as a pupa in a cocoon inside infested seeds. There are two overlapping generations during a year in natural conditions because of the diapause the parasitoid larvae go into at the end of the second generation. Additionally, parasitism activity of *B.* (*G.*) *variator* Nees, 1812, was also observed on an unknown seed feeder (Lepidoptera: Pyralidae). This was the only other host record for *B.* (*G.*) *variator* Nees, 1812. The unknown pyralid larva was green in color with large body, larger than *C. johanssoni* Aarvik and Karsholt, 1993, larva

#### *Lepidopter Parasitoidea DOI: http://dx.doi.org/10.5772/intechopen.86381*

*Trends in Integrated Insect Pest Management*

**Figure 7.**

**Figure 8.**

*feeder larva, Cydia spp. [31].*

*(Hymenoptera: Braconidae). Original.*

idiobiont, whose hosts stop development, when they are parasitized. Idiobionts are either ectoparasitoids that kill their hosts or endoparasitoids that attack immobile host stages such as eggs or pupae. Koinobionts, allow the hosts to continue their development until the parasitoid's offspring matures. Most koinobionts are endoparasitoids of larval stages of insects, although a few are ectoparasitic [27]. Some female parasitoids also use the ovipositor to puncture a host and then feed on the body fluids before selecting other hosts for oviposition, thus causing two different types of mortality in the caterpillar pest population. In some cases, for example, *B.* (*G.*) *variator* Nees, 1812, the egg is laid externally on the body of the host and the larvae also feed externally. This parasitoid wasp is a highly polyphagous gregarious ectoparasitoid that attacks the larvae of a wide range of insects, such as Lepidoptera (**Figure 8**). For example, see [28], *B. (G.) variator* Nees, 1812, paralyzes the larvae of *Hadena bicruris* (Hufnagel, 1766) (Lepidoptera: Noctuidae) before depositing on average 3 eggs on 3rd-5th larval instars [29, 30]. Like *C. johanssoni* Aarvik & Karsholt, 1993, *H. bicruris* (Hufnagel, 1766) is a seed specialist, feeding on *Silene latifolia* Poir. 1789 (Caryophyllaceae) seeds. Lepidoptera larvae, attacks the young seeds before they are shed by the parent plant [32–34]. The larvae of the beech moth, *Cydia fagiglandana* (Zeller, 1841), feed inside the nuts of European beech, *Fagus sylvatica* Linnaeus, 1753 (Fabaceae) causing high seed mortality in South Sweden beech forests [35, 36]. Other economically important species are the pea moth, *C. nigricana* (Fabricius, 1794), attacking legume crops; the spruce seed moth, *C. strobilella* Linnaeus, 1758, attacking

*Images general view of the larval ectoparasitoid wasp, Bracon (Glabrobracon) variator Nees, 1812, (Hymenoptera: Braconidae) and its host larva, Cydia johanssoni Aarvik & Karsholt, 1993 (Lepidoptera: Tortricidae): (A) female, oviposition on infested pod of stinking bean trefoil by pyralid seed feeder larva; (B) parasitoid's eggs ((B. (G.) variator)) laid close to the head of host larva (C. johanssoni); (C) developed parasitoid larvae inside the seed and (D) parasitoid larvae ((B. (G.) variator)) feeding upon Pyralid seed* 

*Image general view of the larval ectoparasitoid wasp, adult female, Bracon (Glabrobracon) variator Nees, 1812* 

**10**

spruce seed, *Picea* spp.; *C. latiferreana* Walsingham, 1879, attacking fruits oak and the hickory shuckworm moth, *C. caryana* Fitch, 1756, an important pest of pecan [37].

First generation adult parasitoids of *B.* (*G.*) *variator* Nees, 1812, appeared in the first ten days of May, with the second generation appearing about a fortnight later, but adult wasps were most abundant, and its percentage parasitism were highest among larvae, in early June (**Table 2**). Adult activity was greatest during the hottest part of the day (12.00–14.00 hours). The parasitoid population was greatest on pods infested with *C. johanssoni* Aarvik and Karsholt, 1993, larvae, early June*.* Oviposition began after a lengthy search period. The female wasp first inspected the drilling into the pod (**Figure 8A**), using her antennae by tapping the pod. She then moved so that her mesothoracic legs straddled the drilling position, lifted her abdomen and inserted her ovipositor almost vertically into the pod (recorded Video film). Almost always each caterpillar had three eggs laid on it (**Figure 8B**); rarely, it was noted that more than 3 eggs were laid up to a maximum of six.

The oviposition is not an easy task. The tip of the ovipositor almost always gets stuck to tiny irregularities of the pod surface. When she loses her balance, re-starts oviposition from the beginning. Since the exact point of drilling is crucial and must be recalculated for accuracy. The eggs were white and bacilli-form, with a diameter approximately equal to the tip of a lab needle (**Figure 8B**). Eggs were oviposited directly onto the host larvae. The incubation period is 18 hours at 33°C and relative humidity of 14%. Upon hatching, the parasitoid larva penetrated the caterpillar's cuticle and fed on the body hemolymph for 4–5 days. Upon seed maturation, the parasitoid maturated larva secreted a webbed cocoon within the seed capsule and then pupated. Adults of second generation leave the seed pod in last ten days of May via a hole made by adult. Adults lived as free-living adults for up to 3–4 days until they mated and oviposited. When the larvae of *C. johanssoni* were paralyzed, their movement became reduced. As a result of feeding by the parasitoid larvae, the moth larva was weakened and eventually is reduced to the head capsule and body cuticle.

The free living adult wasps fed on resin produced when opens the first larval stage of *C. johanssoni* Aarvik and Karsholt, 1993, in the pod. The parasitoid aestivation and hibernation was as a pupa in a cocoon inside infested seeds. There are two overlapping generations during a year in natural conditions because of the diapause the parasitoid larvae go into at the end of the second generation. Additionally, parasitism activity of *B.* (*G.*) *variator* Nees, 1812, was also observed on an unknown seed feeder (Lepidoptera: Pyralidae). This was the only other host record for *B.* (*G.*) *variator* Nees, 1812. The unknown pyralid larva was green in color with large body, larger than *C. johanssoni* Aarvik and Karsholt, 1993, larva and had a very low population in the study area (**Figure 8D**).

The parasitization rate on *C. johanssoni* averaged 18.77% ± 3.80, during second generation of *B.* (*G.*) *variator* Nees, 1812 (**Tables 1** and **2**) [31].

General distribution of this wasp includes: China, Central Asia, Mongolia, Siberia, Russia, Crimea, Iran, Turkey and European country [26].


#### **Table 1.**

*The life stages longevity, release period and sex ratio of Trichogramma brassicae Bezdenko, 1968, reared on the grain moth eggs, Sitotroga cerealella Olivier, 1789 (Lepidoptera: Galleriidae) at 28 ± 2°C.*


#### **Table 2.**

*Total number, number of parasitized larvae and the percentage parasitism of Cydia larvae (Cydia johanssoni Aarvik and Karsholt, 1993 (Lepidoptera: Tortricidae) by the second generation of Bracon (Glabrobracon)*  variator *Nees, 1812 (Hymenoptera: Braconidae), Iran, 2014.*

#### **9. Discussion**

*Trichogramma brassicae* Bezdenko, 1968 (Hymenoptera, Trichogrammatidae) has been the object of great interest regarding its mass rearing and is used as a biocontrol agent against many moth pests, in studied area. This study also confirmed that *B.* (*G.*) *variator* is a gregarious idioboint ectoparasitoid with 2 generation in a year, attacks third-fifth instars *C. johanssoni* Aarvik and Karsholt, 1993, caterpillars. Almost always 3 parasitoid eggs were oviposited on each host larva within the pod, which had been previously paralyzed. These observations agree with those of Elzinga [29]. However, Elzinga has not mention about number of eggs and location of pupation.

Because of proper parasitism rate of *B.* (*G.*) *variator* Nees, 1812, this parasitoid plays important role in decrease population of *C. johanssoni* Aarvik and Karsholt, 1993, larvae and it is the most important natural enemy of *C. johanssoni* Aarvik and Karsholt, 1993, in Iran, therefore with conservation of this wasp would take an important step toward reduction of damage of pest moth and development of Stinking bean trefoil shrub in the west of Iran.

Parasitoid sex ratio: In the field condition, female population of *B.* (*G.*) *variator* Nees, 1812, was much more than male and male population was rare.

*B.* (*G.*) *variator* Nees, 1812, has been recorded from most parts of Iran, in East Azerbaijan [38], South Iran [25], North Central Iran [39] and from western part of Iran [31]. *B.* (*G.*) *variator* Nees, 1812, is already known as a larval parasitoid of the lychnis, *Hadena bicruris*, (Hufnagel, 1766), which is the most important of *Silene* 

**13**

**Author details**

provided the original work is properly cited.

Razi University, Kermanshah, Iran

Hassan-Ali Vahedi\*, Jabbar Valipour and Abbas Ali Zamani

\*Address all correspondence to: vnassah@yahoo.com

*Lepidopter Parasitoidea*

*DOI: http://dx.doi.org/10.5772/intechopen.86381*

*latifolia* Poir. 1789 (Caryophyllaceae). This ectoparasitoid attacks 3rd-5th instars moth larvae and stops host development immediately by paralyzing the caterpillars. *B.* (*G.*) *variator* Nees, 1812, is a gregarious parasitoid, which means that several parasitoid larvae attack a given caterpillar and produces clutches that are predominantly single-sex, mainly female (each individual parasitoid lays predominantly three sexed eggs). Females inject paralyzing venom into the host before oviposition. On hatching, the parasitoid larvae perforate the cuticle and imbibe nutrients from the paralyzed or dead host. The lychnis, *Hadena bicruris*, (Hufnagel, 1766) hibernates as a pupa in a cocoon [29]. The adult wasp *B.* (*G.*) *variator* Nees, 1812, feeds on resin of infested pods in the studied forest habitat, western Iran. *B.* (*G.*) *variator* Nees, 1812, is a widespread species, known from: China, Middle Asia, Mongolia,

Siberia, Russia, Crimea, Iran, Turkey and Europeans countries [40].

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

Department of Plant Protection, Compus of Agriculture and Natural Resources,

### *Lepidopter Parasitoidea DOI: http://dx.doi.org/10.5772/intechopen.86381*

*Trends in Integrated Insect Pest Management*

*C. johanssoni* **larvae**

**Date Total number of** 

1 June 2014

2 June 2014

3 June 2014

4 June 2014

5 June 2014

6 June 2014

7 June 2014

8 June 2014

9 June 2014

10 June 2014

**Table 2.**

**12**

**9. Discussion**

of pupation.

Stinking bean trefoil shrub in the west of Iran.

variator *Nees, 1812 (Hymenoptera: Braconidae), Iran, 2014.*

*Trichogramma brassicae* Bezdenko, 1968 (Hymenoptera, Trichogrammatidae) has been the object of great interest regarding its mass rearing and is used as a biocontrol agent against many moth pests, in studied area. This study also confirmed that *B.* (*G.*) *variator* is a gregarious idioboint ectoparasitoid with 2 generation in a year, attacks third-fifth instars *C. johanssoni* Aarvik and Karsholt, 1993, caterpillars. Almost always 3 parasitoid eggs were oviposited on each host larva within the pod, which had been previously paralyzed. These observations agree with those of Elzinga [29]. However, Elzinga has not mention about number of eggs and location

Mean 116 21.35 18.77 ± 3.80

*Total number, number of parasitized larvae and the percentage parasitism of Cydia larvae (Cydia johanssoni Aarvik and Karsholt, 1993 (Lepidoptera: Tortricidae) by the second generation of Bracon (Glabrobracon)* 

**Number of parasitized larvae of** *C. johanssoni* **by** *B. variator*

120 9 7.5

105 7.5 7.14

120 9 7.5

120 7 5.83

135 22 16.29

110 20 18.18

130 29 22.3

110 38 34.54

100 33 33

110 39 35.45

**Parasitism rate (%)**

Because of proper parasitism rate of *B.* (*G.*) *variator* Nees, 1812, this parasitoid plays important role in decrease population of *C. johanssoni* Aarvik and Karsholt, 1993, larvae and it is the most important natural enemy of *C. johanssoni* Aarvik and Karsholt, 1993, in Iran, therefore with conservation of this wasp would take an important step toward reduction of damage of pest moth and development of

Parasitoid sex ratio: In the field condition, female population of *B.* (*G.*) *variator*

*B.* (*G.*) *variator* Nees, 1812, has been recorded from most parts of Iran, in East Azerbaijan [38], South Iran [25], North Central Iran [39] and from western part of Iran [31]. *B.* (*G.*) *variator* Nees, 1812, is already known as a larval parasitoid of the lychnis, *Hadena bicruris*, (Hufnagel, 1766), which is the most important of *Silene* 

Nees, 1812, was much more than male and male population was rare.

*latifolia* Poir. 1789 (Caryophyllaceae). This ectoparasitoid attacks 3rd-5th instars moth larvae and stops host development immediately by paralyzing the caterpillars. *B.* (*G.*) *variator* Nees, 1812, is a gregarious parasitoid, which means that several parasitoid larvae attack a given caterpillar and produces clutches that are predominantly single-sex, mainly female (each individual parasitoid lays predominantly three sexed eggs). Females inject paralyzing venom into the host before oviposition. On hatching, the parasitoid larvae perforate the cuticle and imbibe nutrients from the paralyzed or dead host. The lychnis, *Hadena bicruris*, (Hufnagel, 1766) hibernates as a pupa in a cocoon [29]. The adult wasp *B.* (*G.*) *variator* Nees, 1812, feeds on resin of infested pods in the studied forest habitat, western Iran. *B.* (*G.*) *variator* Nees, 1812, is a widespread species, known from: China, Middle Asia, Mongolia, Siberia, Russia, Crimea, Iran, Turkey and Europeans countries [40].

## **Author details**

Hassan-Ali Vahedi\*, Jabbar Valipour and Abbas Ali Zamani Department of Plant Protection, Compus of Agriculture and Natural Resources, Razi University, Kermanshah, Iran

\*Address all correspondence to: vnassah@yahoo.com

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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[2] Noyes JS, Valentine EW. Chalcidoidea (Insecta: Hymenoptera)—Introduction, and review of genera in smaller families. Fauna of New Zealand. 1989;**18**:1-91

[3] Walter S. The biology and ecology of egg parasites of the genus *Trichogramma* Westwood (Hym., Chalc.). Part I.: Studies in selected forest biocoenoses in the G.D.R. Zoologische Jahrbücher, Abteilung für Systeamtik, Ökologie und Geographie der Tiere. 1983;**110**(3):271-299 (in German)

[4] Walter S. Biological and ecological studies on egg parasites of the genus *Trichogramma* Westwood (Hym. Chalc.). Part 2. Investigations carried out under laboratory conditions. Zoologische Jahrbücher, Anteilung für Systematik, Ökologie und Geographie der Tiere. 1983;**110**(4):419-441 (in German)

[5] Knutson A. The Trichogramma Manual: A Guide to the Use of Trichogramma for Biological Control with Special Reference to Augmentative Releases for Control of Bollworm and Budworm in Cotton B-6071. College Station, TX: Texas Agriculture Extension Service, Texas A&M University System; 1998

[6] Metcalfe JR, Brenière J. Egg parasites (*Trichogramma* spp.) for control of sugar cane moth borers. In: Williams JR, Metcalfe RW, Mungomery RW, Mathes R, editors. Pests of Sugar-Cane. Amsterdam: Elsevier Publishing Company; 1969. pp. 81-116

[7] Ruberson JR, Kring TJ. Parasitism of developing eggs by *Trichogramma pretiosum* (Hymenoptera:

Trichogrammatidae): Host age preference and suitability. Biological Control. 1993;**3**:39-46

[8] Jozian A, Baigi S. Efficiency control of parasitoid, *Bracon hebetor* (Say) (Hymenoptera: Braconidae) on the chickpea pests. In: 5th Iranian Pulses Symposium, Ilam Province; 2012. Available from: https://www.civilica. com/Paper-PULSES05-PULSES05\_028. html.COI:PULSES05\_028

[9] Salehnia N. Biocontrol of the pomegranate calyx worm, *Ectomyelois ceratoniae* (Lepidoptera, Pyralidae) in pomegranate orchards. In: 4th National Symposium on Pomegranate, Iran; 2011. Ferdos: Khorasan. Available from: https://www.civilica.com/ Paper-ANAR01-ANAR01\_083.html [in Persian]

[10] Sevcan O. Natural parasitism rate of *Trichogramma evanescens* Westwood (Hymenoptera: Trichogrammatidae) and its release efficacy against the cotton bollworm, *Helicoverpa armigera* Hübner (Lepidoptera: Noctuidae) in the Cukurova region Turkey. Entomological News. 2008;**119**(1). DOI: 10.3157/0013-872X(2008)119[19,NPRO TE]2.0.CO;2

[11] Rabieh MM. Biodiversity of noctuid moths (Lepidoptera: Noctuidae) in the agroecosystems of Mashhad County. Biodiversity International Journal. 2018;**2**(2):147-151. DOI: 10.15406/ bij.2018.02.00057

[12] Mehrnejad MR. The current status of pistachio pests in Iran. In: Ak BE, editor. XI GREMPA Seminar on Pistachios and Almonds. CIHEAM. 2001;**56**:315-322

[13] Mehrnejad MR. The carob moth, a pest of pistachio nut in Iran. Acta Horticulturae. 1995;**419**:365-372

**15**

*Lepidopter Parasitoidea*

*DOI: http://dx.doi.org/10.5772/intechopen.86381*

[21] Ward DF. Braconidae of New Zealand. 2014. Available from: http:// braconidae.landcareresearch.co.nz

[22] Quicke DLJ. The Braconid and Ichneumonid Parasitoid Wasps: Biology, Systematics, Evolution and Ecology. Oxford, UK: Wiley-Blackwell; 2015.

[23] Belokobylskij SA, Tobias VI,

Kotenko AG, Proshchalykin M, Yu DSK. 48. Fam. Braconidae – Braconidy. In: Lelej AS, editor. Annotated Catalogue of Insects of the Russian Far East. Vol. I Hymenoptera. Vladivostok: Dal'nauka;

[24] Broad GR, Shaw MR, Godfray HC. Checklist of British and Irish Braconidae (Hymenoptera). 2012. Available from: http://www.nhm.ac.uk/resources-rx/ files/braconidae-checklist-for-

[25] Ameri A, Talebi AA, Beyarslan A, Kamali K, Rakhshani E. Study of the genus *Bracon* Fabricius, 1804 (Hymenoptera: Braconidae) of

Southern Iran with description of a new species. Zootaxa. 2013;**3754**(4):353-380

Horstmann K. World Ichneumonoidea 2004. Taxonomy, biology, morphology and distribution, Taxapad 2005 (Scientific names for information management), interactive catalogue on DVD/CDROM; Canada: Vancouver;

[27] Sharkey MJ. Family Braconidae, in Hymenoptera of the world: An identification guide to families. In: Goulet H, Huber JT, editors. Ottawa, Canada: Research Branch, Agriculture Canada Publications; 1993. pp. 362-395

[28] Quicke DLJ, van Achterberg C. Phylogeny of the subfamilies of the family Braconidae (Hymenoptera:

[26] Yu DS, van Achterberg C,

[Accessed: 15 March 2019]

704 pp

2012. pp. 300-389

web-34139.pdf

2012

[14] Vahedi HA. Mass production of insect pests' natural enemies as biological control agents

(Trichogramma and Cryptolemus). Unpublished manuscript. Kermanshah, Iran: Department of Plant Protection, Campus of Agriculture and Natural Resources, Razi University; 2012; 30 pp

[15] Bigler F. Quality control in Trichogramma production. In: Wajnberg E, Hassan SA, editors. Biological Control with Egg Parasitoids. Oxon, UK: CAB International; 1994.

[16] Vasquez LA, Shelton AM, Hoffmann MP, Roush RT. Laboratory evaulation of commercial Trichogrammatid products for potential use against *Plutella xylostella* (L.) (Lepidoptera: Plutellidae). Biological Control.

[17] Hassan SA. Strategies to select *Trichogramma* species for use in biological control. In: Wajnberg E, Hassan SA, editors. Biological Control with Egg Parasitoids. Oxon, UK: CAB

International; 1994. pp. 55-71

[18] Nagaraja H. Mass production of Trichogrammatid parasitoids. In: Sithanantham S, Ballal C, Jalali S, Bakthavatsalam N, editors. Biological Control of Insect Pests Using Egg Parasitoids. New Delhi: Springer; 2013

[19] Lachaud JP, Gabriela PL. Diversity

Hymenopteran parasitoids of ants: A review. Psyche. 2012;**2012**:1-24. Article

Classification and biology of braconid wasps (Hymenoptera: Braconidae). In: Dowling WR, Askew RR, editors. Handbooks for the Identification of British Insects, Part 11. Vol. 7. London, UK: Royal Entomological Society of

of species and behavior of

[20] Shaw MR, Huddleston T.

London; 1991. pp. 1-126

ID 134746

pp. 93-111

1997;**9**:143-148

*Lepidopter Parasitoidea DOI: http://dx.doi.org/10.5772/intechopen.86381*

[14] Vahedi HA. Mass production of insect pests' natural enemies as biological control agents (Trichogramma and Cryptolemus). Unpublished manuscript. Kermanshah, Iran: Department of Plant Protection, Campus of Agriculture and Natural Resources, Razi University; 2012; 30 pp

[15] Bigler F. Quality control in Trichogramma production. In: Wajnberg E, Hassan SA, editors. Biological Control with Egg Parasitoids. Oxon, UK: CAB International; 1994. pp. 93-111

[16] Vasquez LA, Shelton AM, Hoffmann MP, Roush RT. Laboratory evaulation of commercial Trichogrammatid products for potential use against *Plutella xylostella* (L.) (Lepidoptera: Plutellidae). Biological Control. 1997;**9**:143-148

[17] Hassan SA. Strategies to select *Trichogramma* species for use in biological control. In: Wajnberg E, Hassan SA, editors. Biological Control with Egg Parasitoids. Oxon, UK: CAB International; 1994. pp. 55-71

[18] Nagaraja H. Mass production of Trichogrammatid parasitoids. In: Sithanantham S, Ballal C, Jalali S, Bakthavatsalam N, editors. Biological Control of Insect Pests Using Egg Parasitoids. New Delhi: Springer; 2013

[19] Lachaud JP, Gabriela PL. Diversity of species and behavior of Hymenopteran parasitoids of ants: A review. Psyche. 2012;**2012**:1-24. Article ID 134746

[20] Shaw MR, Huddleston T. Classification and biology of braconid wasps (Hymenoptera: Braconidae). In: Dowling WR, Askew RR, editors. Handbooks for the Identification of British Insects, Part 11. Vol. 7. London, UK: Royal Entomological Society of London; 1991. pp. 1-126

[21] Ward DF. Braconidae of New Zealand. 2014. Available from: http:// braconidae.landcareresearch.co.nz [Accessed: 15 March 2019]

[22] Quicke DLJ. The Braconid and Ichneumonid Parasitoid Wasps: Biology, Systematics, Evolution and Ecology. Oxford, UK: Wiley-Blackwell; 2015. 704 pp

[23] Belokobylskij SA, Tobias VI, Kotenko AG, Proshchalykin M, Yu DSK. 48. Fam. Braconidae – Braconidy. In: Lelej AS, editor. Annotated Catalogue of Insects of the Russian Far East. Vol. I Hymenoptera. Vladivostok: Dal'nauka; 2012. pp. 300-389

[24] Broad GR, Shaw MR, Godfray HC. Checklist of British and Irish Braconidae (Hymenoptera). 2012. Available from: http://www.nhm.ac.uk/resources-rx/ files/braconidae-checklist-forweb-34139.pdf

[25] Ameri A, Talebi AA, Beyarslan A, Kamali K, Rakhshani E. Study of the genus *Bracon* Fabricius, 1804 (Hymenoptera: Braconidae) of Southern Iran with description of a new species. Zootaxa. 2013;**3754**(4):353-380

[26] Yu DS, van Achterberg C, Horstmann K. World Ichneumonoidea 2004. Taxonomy, biology, morphology and distribution, Taxapad 2005 (Scientific names for information management), interactive catalogue on DVD/CDROM; Canada: Vancouver; 2012

[27] Sharkey MJ. Family Braconidae, in Hymenoptera of the world: An identification guide to families. In: Goulet H, Huber JT, editors. Ottawa, Canada: Research Branch, Agriculture Canada Publications; 1993. pp. 362-395

[28] Quicke DLJ, van Achterberg C. Phylogeny of the subfamilies of the family Braconidae (Hymenoptera:

**14**

*Trends in Integrated Insect Pest Management*

[1] Pennachio F. Strand MR evolution of developmental strategies in parasitic Hymenoptera. Annual Review of Entomology. 2006;**51**:233-258

Trichogrammatidae): Host age preference and suitability. Biological

[8] Jozian A, Baigi S. Efficiency control of parasitoid, *Bracon hebetor* (Say) (Hymenoptera: Braconidae) on the chickpea pests. In: 5th Iranian Pulses Symposium, Ilam Province; 2012. Available from: https://www.civilica. com/Paper-PULSES05-PULSES05\_028.

Control. 1993;**3**:39-46

html.COI:PULSES05\_028

Persian]

TE]2.0.CO;2

bij.2018.02.00057

2001;**56**:315-322

[9] Salehnia N. Biocontrol of the pomegranate calyx worm, *Ectomyelois ceratoniae* (Lepidoptera, Pyralidae) in pomegranate orchards. In: 4th National Symposium on Pomegranate, Iran; 2011. Ferdos: Khorasan. Available from: https://www.civilica.com/ Paper-ANAR01-ANAR01\_083.html [in

[10] Sevcan O. Natural parasitism rate of *Trichogramma evanescens* Westwood (Hymenoptera: Trichogrammatidae) and its release efficacy against the cotton bollworm, *Helicoverpa armigera* Hübner (Lepidoptera: Noctuidae) in the Cukurova region Turkey.

Entomological News. 2008;**119**(1). DOI: 10.3157/0013-872X(2008)119[19,NPRO

[11] Rabieh MM. Biodiversity of noctuid moths (Lepidoptera: Noctuidae) in the agroecosystems of Mashhad County. Biodiversity International Journal. 2018;**2**(2):147-151. DOI: 10.15406/

[12] Mehrnejad MR. The current status of pistachio pests in Iran. In: Ak BE, editor. XI GREMPA Seminar on Pistachios and Almonds. CIHEAM.

[13] Mehrnejad MR. The carob moth, a pest of pistachio nut in Iran. Acta Horticulturae. 1995;**419**:365-372

[2] Noyes JS, Valentine EW. Chalcidoidea (Insecta: Hymenoptera)—Introduction, and review of genera in smaller families. Fauna of New Zealand. 1989;**18**:1-91

[3] Walter S. The biology and ecology of egg parasites of the genus *Trichogramma* Westwood (Hym., Chalc.). Part I.: Studies in selected forest biocoenoses in the G.D.R. Zoologische Jahrbücher, Abteilung für Systeamtik, Ökologie und Geographie

[4] Walter S. Biological and ecological studies on egg parasites of the genus *Trichogramma* Westwood (Hym. Chalc.). Part 2. Investigations carried out under laboratory conditions. Zoologische Jahrbücher, Anteilung für Systematik, Ökologie und Geographie der Tiere. 1983;**110**(4):419-441

[5] Knutson A. The Trichogramma Manual: A Guide to the Use of Trichogramma for Biological Control with Special Reference to Augmentative Releases for Control of Bollworm and Budworm in Cotton B-6071. College Station, TX: Texas Agriculture Extension Service, Texas A&M

[6] Metcalfe JR, Brenière J. Egg parasites (*Trichogramma* spp.) for control of sugar cane moth borers. In: Williams JR, Metcalfe RW, Mungomery RW, Mathes

R, editors. Pests of Sugar-Cane. Amsterdam: Elsevier Publishing Company; 1969. pp. 81-116

*pretiosum* (Hymenoptera:

[7] Ruberson JR, Kring TJ. Parasitism of developing eggs by *Trichogramma* 

University System; 1998

der Tiere. 1983;**110**(3):271-299

(in German)

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[31] Valipour J, Vahedi HA, Zamani AA. An outline on biology and behavior of *Bracon variator* Nees, 1812 (Hym.: Braconidae), an ectoparasitoid of *Cydia johanssoni* Aarvik and Karsholt, 1993 (Lepidoptera: Tortricidae) from Iran. Biharean. Biologist. 2017;**11**(1):15-19

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**17**

Section 2

Insecticides
