Development of an artificial host egg for in vitro egg laying of Trichogramma galloi and T. pretiosum using plastic membranes

An artificial host egg was developed as a first attempt to improve the techniques for in vitro rearing of Trichogramma galloi Zucchi and T. pretiosum Riley (Hymenoptera, Trichogrammatidae). Plastic membranes (polyethylene, polypropylene and polystyrene) of different thickness were tested for their usefulness in manufacturing artificial eggs, using a thermal system to produce semi-spherical cupules on the plastic surface. These artificial eggs were filled with larval hemolymph of Helicoverpa zea (Boddie) (Lepidoptera, Noctuidae), and the optimal size of the artificial egg and density of females/artificial egg for the highest host acceptance (percentage of artificial eggs with at least one parasitoid egg) as well as the ideal level of parasitism (eggs/artificial egg) were determined. High-density polyethylene 7–8 and 9–10 m thick resulted in the best acceptance and level of parasitization for both parasitoid species. Six females per artificial egg resulted in the best rate of parasitization for these parasitoids, although these data were not different when using 4 or 5 females of T. galloi. The size, shape and surface texture of the artificial eggs were found to provide enough cues to elicit parasitization behavior in Trichogramma females, since no chemical stimulus was provided.     

卵液成分改变及卵表涂施引诱剂对玉米螟赤眼蜂产卵发育的影响

试验通过部分改变人造卵卵液成分和在人造卵表涂施引诱剂2种途径来试图提高人造卵繁育玉米螟赤眼蜂Trichogramma ostriniae Panget Chen的成功率。结果表明,不同比例的亚洲玉米螟OstriniafurnacalisGuenée幼虫和蛹匀浆替代部分柞蚕蛹匀浆后,对赤眼蜂寄生率有明显提高,但赤眼蜂在人造卵内的化蛹率和羽化率均较低,赤眼蜂发育至老熟幼虫因卵液过剩而被淹死;在原卵液卵表涂施亚洲玉米螟卵和鳞片正己烷提取液后,评价繁蜂成功率的各项参数都有明显提高,赤眼蜂可以完成发育,羽化成蜂,但繁蜂成功率还较低。综合上述试验结果,应用人造卵繁殖玉米螟赤眼蜂研究重点要解决卵液过剩以及其它相关的营养学问题。     

The potential of trehalose to replace insect hemolymph in artificial media for Trichogramma dendrolimi (Hymenoptera: Trichogrammatidae)

Insect additives have been shown to improve the value of artificial media for Trichogramma species, but at the same time maintain dependence on parallel cultures of host insects. In the present study, Trichogramma dendrolimi Matsumura was reared in vitro from egg to adult on artificial media with different contents of pupal hemolymph of Chinese oak silkmoth Antheraea pernyi (Guérin‐Méneville) and with supplements of distilled water, or of trehalose dissolved in water or in Grace's insect medium. The results indicated insect hemolymph was the key component of artificial medium. Developmental parameters, including rates of parasitism, pupation, adult emergence and normal adults, and numbers of produced adults, were increased on media supplemented with trehalose even when the proportion of pupal hemolymph was reduced. Two artificial media, the first containing 30% hemolymph and 10% trehalose in water with 98.9% parasitism rate, 77.7.0% pupation rate, 77.2% emergence rate, 80.0% normal adult rate and 333 produced adults, and the second containing 25% hemolymph and 15% trehalose in Grace's insect medium with 97.8% parasitism rate, 91.0% pupation rate, 85.2% emergence rate, 76.1% normal adult rate and 757 produced adults, were believed to hold potential to mass produce T. dendrolimi. The use of trehalose to partially replace pupal hemolymph in artificial medium of this and other Trichogramma species may contribute to a significant reduction in their production cost and may as such help to evade problems related to short supplies of lepidopteran eggs, which currently constitute the main factitious host for the mass rearing of the parasitoids.     

First report of parasitism of the Trichogramma bruni Nagaraja (Hymenoptera: Trichogrammatidae) in eggs of urbanus proteus (L.) (Lepidoptera: Hesperiidae) in snap beans (Phaseolus vulgaris) ( L.)

Aiming to verify the presence of eggs parasitoids of the genus Trichogramma, for using in the biological control of insects pests, were collected eggs from long tailed skipper butterfly, a lepidopteran defoliator of snap beans, which presented eggs with parasitism characteristics (dark eggs). The eggs were taken to the laboratory were 44.9% of parasitism was verified. Viability was 96.3%, with an average of 6.6 individuals per egg and a sexual ratio of 0.93. Male specimens were identified as Trichogramma bruni Nagaraja. It is the first report of parasitism in this host and culture.     

东北地区四种本地赤眼蜂对草地贪夜蛾寄生能力及适应性研究

草地贪夜蛾Spodoptera frugiperda(Smith)是新入侵我国的重大危险性入侵害虫,赤眼蜂是我国生物防治的优势卵寄生蜂,可用于防治多种农林作物上的鳞翅目害虫。为检测东北地区本地赤眼蜂对草地贪夜蛾卵的生防潜能,筛选防治草地贪夜蛾的有效蜂种,本研究比较研究了松毛虫赤眼蜂、玉米螟赤眼蜂、黏虫赤眼蜂和稻螟赤眼蜂对草地贪夜蛾卵的寄生能力和发育适合度。结果表明,4种赤眼蜂均能寄生草地贪夜蛾卵,并完成发育,但其寄生能力和发育适合度存在显著差异。松毛虫赤眼蜂、玉米螟赤眼蜂和黏虫赤眼蜂对草地贪夜蛾卵块寄生率均高于80%,但仅松毛虫赤眼蜂对草地贪夜蛾卵表现出较高的卵粒寄生量。松毛虫赤眼蜂、玉米螟赤眼蜂和黏虫赤眼蜂子代羽化率和雌性比无显著差异,但松毛虫赤眼蜂比其它3种赤眼蜂的发育历期更短,而稻螟赤眼蜂对草地贪夜蛾卵的寄生能力和适合度均最差。研究结果将为应用本地赤眼蜂生物防治潜在入侵东北地区的草地贪夜蛾提供技术支撑。     

Trichogramma parasitoids alter the metabolic physiology of Manduca eggs

Egg parasitoids face unique developmental constraints. First, they have exceptionally limited resources to support themselves and their siblings through three life stages. Second, they develop within the physiological system of another species, which they modify to their own ends. We examined how these constraints affect the metabolic physiology of egg parasitism, and whether parasitoids retool their host eggshell to account for their different metabolic demands. Higher-conductance eggshells allow more oxygen to reach the developing parasitoids, but also allow more water to leave the egg. We used Manduca sexta (Lepidoptera: Sphingidae) eggs and Trichogramma (Hymenoptera: Trichogrammatidae) parasitoids from southeastern AZ, USA. Compared with unparasitized Manduca eggs, eggs parasitized by Trichogramma had lower peak metabolic rates and approximately 50 per cent lower metabolic efficiency. However, developing Trichogramma were far more efficient than typical transfer efficiencies between tropic levels (approx. 10%). Even within a few hours of parasitization, eggs containing more Trichogramma had lower per-parasitoid metabolic rates, suggesting that parasitoid larvae have mechanisms for rapidly adjusting their metabolic rates based on number of siblings. Parasitoids also appear to control the conductance of their host eggshell: their different metabolic demands were mirrored by shifts in rates of water loss.     

松毛虫卵期几种寄生蜂的共寄生现象及其对寄生率的影响

松毛虫赤眼蜂Ｔｒｉｃｈｏｇｒａｍｍａｄｅｎｄｒｏｌｉｍ　Ｍａｔｓｕｍｕｒａ,白跗平腹小蜂ＰｓｅｕｄａｎａｓｔａｔｕｓａｌｂｉｔａｒｓｉｓＡｓｈｍｅａｄ,松毛虫宽缘金小蜂Ｐａｃｈｙｎｅｕｒｏｎ　ｓｏｌｉｔａｒｉｕｍ（Ｈａｒｔｉｇ）,松毛虫黑卵蜂Ｔｅｌｅｎｏ－ｍｕｓ　ｄｅｎｄｒｏｌｉｍｕｓｉ　Ｃｈｕ和大蛾卵跳小蜂Ｏｏｅｎｃｙｒｔｕｓｋｕｗａｎａｅ（Ｈｏｗａｒｄ）是松毛虫卵期几种主要寄生蜂。通过对上述几种寄生蜂寄生习性的研究表明：赤眼蜂和平腹小蜂不但能寄生新鲜害虫卵,还可以寄生对方已寄生并已发育１—３天的寄生卵,但羽化率均不高;松毛虫宽缘金小峰不寄生新鲜害虫卵,专门寄生赤眼蜂寄生后已发育１—７天的寄生卵,其中又以赤眼蜂发育３—５天的卵寄生率和羽化率为最高。除赤眼蜂和平腹小蜂可以混合使用外,其它蜂种混用都不同程度地存在相互排挤的现象。如果先让平腹小蜂寄生尔后再让赤眼蜂寄生或先让赤眼蜂寄生,尔后再让跳小蜂或黑卵蜂寄生,能够提高卵块寄生率和充分发挥各自天敌的作用。     

Constraints on the use of Trichogramma Egg Parasitoids in Biological Control Programmes in India

The use of Trichogramma (Hymenoptera: Trichogrammatidae) egg parasitoids for the biological control of several insect pests, including the pod borer Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae), is being promoted in India. Several companies supply these parasitoids to farmers, but no evaluation of product quality has been made. Low product quality is recognized as being one of the most important reasons for the failure of these biocontrol agents. In this study, Trichogramma chilonis Ishii egg-cards were obtained from eight producers in southern India. Product quality was evaluated using the following parameters: number of eggs/card, percentage parasitism, percentage emergence, progeny sex ratio and percentage of deformed females. In addition, producers were asked for the average values of these parameters in their production systems and for their recommendations for the use of T. chilonis to control H. armigera in cotton. The quality of the egg-cards was highly variable and was much lower than that stated by the producers. The recommended frequency and interval of releases, and the number of parasitoids to be released, differed greatly among producers. The low product quality results in up to 85% fewer parasitoids being released than is recommended by the producer. The effect of the low product quality and the highly variable release recommendations on applied biological control programmes is discussed. Suggestions for more effective use of Trichogramma spp. are given.     

Possible incompatibility between transgenic cottons and parasitoids

Abstract  We investigated the rates of egg and larval parasitism in transgenic and non-transgenic, conventional cottons. Sentinel eggs and larvae of the cotton bollworm, CBW, Helicoverpa armigera Hübner, were released and collected at regular intervals across the cotton growing season, and the relationship between parasitism and different cotton cultivars determined. Egg and larval parasitism were significantly lower in the transgenic cottons than in the non-transgenic conventional cottons. The egg parasitoid recovered was Trichogramma confusum Viggiani and the predominant larval parasitoids were Campoletis chlorideae Uchida and Meteorus pulchriconis (Wesmael). Our studies indicate a potential negative interaction between transgenic cottons and parasitoids of CBW but need to be interpreted with caution because no within-year replication was used and treatments were not spatially randomised across years.     

松毛虫卵期几种寄生蜂的共寄生现象及其对寄生率的影响

松毛虫赤跟蜂 Trichogramma dendrolimi Matsumura 白跗平腹Pseudanastatusalbitarsis Ashmead松毛虫宽缘金小蜂Pachyneuron olitarium(Harig), 松毛虫黑卵蜂 Telenomus dendrolimusi Chu和大蛾卵跳小蜂Ooenvyrtus Kuwanae(Howard)是松毛虫卵期几种主要寄生蜂。通过对上述几种寄生蜂寄生习性的研究表明：赤眼蜂和平腹小蜂不但能寄生新鲜害虫卵,还可以寄生对方已寄生并巳发育1-3天的寄生卵,但羽化串均不高;松毛虫宽缘金小蜂不寄生新鲜害虫卵,专门寄生赤眼蜂寄生后巳发育1-7,天的寄生卵,其中又以赤眼蜂发育3-5天的卵寄生串和羽化率为最高。除赤眼蜂和平腹小蜂可以混合使用外,其它蜂种混用都不同程度地存在相互排挤的现象。如果先让平腹小蜂寄生尔后再让赤眼蜂寄生或先让赤眼蜂寄生,尔后再让跳小蜂或黑卵蜂寄生,能够提高卵块寄生串和充分发挥各自天敌的作用。     

Estimating parasitoid immature mortality by comparing oviposition and pupal development of Trichogramma galloi Zucchi and T. pretiosum Riley on natural and factitious hosts

Mortality during the immature development of T. galloi and T. pretiosum was estimated on UV-killed and live eggs of a factitious and a natural host, respectively. A staining technique was used to determine the actual parasitization of UV-treated eggs and was compared with the number of parasitoids that emerged per host egg (detectable parasitization). Effects of temperature as a factor of mortality during the immature development of both parasitoids on the factitious host was also assessed. The actual and detectable parasitization of live hosts was measured by recording both the parasitization behavior and the number of eggs where a parasitoid developed successfully. Our data show that mortality during immature development of both parasitoids may occur in live eggs of the natural host. No such mortality was observed when parasitoids developed on UV-killed eggs of the factitious host. Possible causes of parasitoid immature mortality and the effects of using UV-treated eggs of factitious hosts in estimating the parasitism capacity of Trichogramma in field conditions are discussed.     

Structure of egg-batches of Thaumetopoea pityocampa (Den. and Schiff.) (Lep., Thaumetopoeidae), egg parasitoids and rate of egg parasitism on the Iberian Peninsula

In four regions of the Iberian Peninsula 273 egg-batches of the pine processionary moth were collected after hatching of the caterpillars in 1991 and 1992. After collection, the batches were stored singly in test tubes closed with foam stoppers under laboratory conditions at 20–25°C. The number of eggs in each egg-batch varied between 37 and 312 and the length of the batches between 0.6 and 4.2 cm. Independently of the pine species infested, seven to nine rows of eggs were counted, wrapped around one pair of needles. A correlation was found between number of eggs and the product of number of egg rows and length of egg-batches (82% <  r ²  < 97%). Oviposition almost always started from the base towards the tip of the needles. The egg parasitoids that emerged were counted daily, removed and identified. Later in the year of oviposition, after the end of the period of emergence of the parasitoids, the scale cover of the egg-batches were removed. One year later, all eggs were opened and the contents, such as parasitoid meconia and remains of caterpillars, were determined. The hatching rate of the caterpillars varied from 47.2 to 79.7%. The rate of parasitism fluctuated between 11.3 and 31.7%, but 44% of parasitoids died before successful emergence. Four species of Chalcidoidea were identified: Baryscapus servadeii (Dom.), Ooencyrtus pityocampae (Mercet), Anastatus bifasciatus (Fonsc.) and Trichogramma sp. Baryscapus servadeii and O. pityocampae were the most abundant species, followed by Trichogramma sp. Baryscapus servadeii emerged mainly in June and O. pityocampae in April or June, depending on the region. For A. bifasciatus only males, and for B. servadeii females emerged except one male. In O. pityocampae , a relatively high percentage of males (5.4–10.4%) was found.     

Host-foraging success of three species of Trichogramma (Hymenoptera: Trichogrammatidae) in a simulated retail environment

Three species of trichogrammatid egg parasitoids (Trichogramma deion Pinto & Oatman, Trichogramma ostriniae Pang & Chen, and Trichogramma pretiosum Riley) (Hymenoptera: Trichogrammatidae) were evaluated under laboratory conditions as potential biological control agents for the Indianmeal moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae), on retail shelves. A single shelving unit was used in each trial and a grid of sentinel egg disks was used to evaluate foraging success. The shelving consisted of pallet units with five shelves that were either bare or stocked with empty cereal boxes. In each replicate, approximately 500 female Trichogramma were released at the center of the shelving unit and allowed to forage for 48 h. Percentage of egg parasitism and percentage of host egg mortality were recorded after 7 d. Foraging success as well as the spatial pattern of parasitism differed significantly among the three Trichogramma species. Percentage of egg parasitism was approximately 4 times greater for T. deion than for T. ostriniae or T. pretiosum. The vertical distribution of parasitism by T. deion was also more uniform than for the other two species. In addition, the presence of packaging affected the foraging efficiency of T. ostriniae and T. pretiosum but not T. deion. Based on these findings, Trichogramma deion may be the best-suited candidate for augmentative biological control of P. interpunctella in retail stores, and a central release point of T. deion will likely provide adequate coverage of products on pallet-type shelving.     

Effect of induced tolerance to Bt toxin on the egg size of Helicoverpa armigera and parasitism by Trichogramma pretiosum

Larvae of Helicoverpa armigera (Hübner) can develop a form of Bt tolerance after exposure to sub‐lethal doses of Bt‐toxin subclass Cry1Ac. Increasing levels of tolerance are produced over generations of larval exposure, which are not related to DNA sequence changes, and are largely maternally transmitted. The characteristic of maternal transmission, combined with the importance of egg parasitoids to cotton pest management, raises questions about the effects of Bt tolerance/exposure on the eggs of H. armigera and on some key metrics of egg parasitism. In the present study, the effect of inducible tolerance on eggs of H. armigera and parasitism by Trichogramma pretiosum (Riley) is investigated. First, the volumes of eggs laid by susceptible and tolerant H. armigera females are compared. In addition, the effect of inducible tolerance on egg parasitism is determined by comparing parasitism success, the number of adult wasps emerged per host egg, and the proportion of male and female offspring emerged per host egg. The results obtained suggest that Cry1Ac‐tolerance is associated with increased egg volume, even after just one generation of sub‐lethal exposure. When tolerant H. armigera are freed from ongoing sub‐lethal exposure, a corresponding decrease in egg volume is not detected. Although there is no difference in the percentage of eggs parasitized, there is an increase in the number of emergent parasitoids, especially males, from eggs laid by tolerant H. armigera. These results confirm that maternally‐transmitted Bt tolerance is reflected in the phenotype of the eggs of tolerant offspring, which affects egg parasitism.     

Acceptability and suitability of Tuta absoluta eggs from irradiated parents to parasitism by Trichogramma nerudai and Trichogramma pretiosum (Hymenoptera: Trichogrammatidae)

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Aerodynamic separation of parasitised from unparasitised insect host eggs

Aerodynamic sorting of host eggs exposed to Trichogramma egg parasitoids improved the percentage parasitism of retained product. Losses of parasitised eggs declined as percentage parasitism of the unsorted eggs increased. Trichogramma eclosion from parasitised eggs was similar for the unsorted and retained fractions; emergence from the parasitised culls was significantly lower.     

小菜蛾卵寄生蜂在生态恢复条件下的自然增长

在生态恢复的深圳龙岗生态示范农场,自然寄生于小菜蛾卵的寄生蜂种类逐渐年多1999年9月,首次发现卷蛾分索赤眼蜂（Trichogrammatoidea bactrae)和玉米螟赤眼蜂（Trichogramma ostriniae)自然寄生于田间小菜蛾卵;同时,拟澳洲赤眼蜂（Tichogramma confusum)和碧岭赤眼蜂（Trichogramma bilingensis)在田间也有一定的自然寄生比率。目前,4种赤眼蜂已经在田间自然建立了种群,其中以拟澳洲赤眼蜂数量占优势小菜蛾的自然寄生率为15％,最高时可达29％。表明农田生态条件的恢复对小菜蛾卵寄生蜂有较好的增殖和保护作用,最后提出了在该农场利用赤眼蜂自然控制小菜蛾种群的可能性。     

Effect of different cold storage periods on parasitization performance of Trichogramma cacoeciae (Hymenoptera,Trichogrammatidae) on eggs of Ephestia kuehniella (Lepidoptera,Pyralidae)

The parasitism rates by Trichogramma cacoeciae Marchal (Hymenoptera, Trichogrammatidae) using Ephestia kuehniella Zell. (Lepidoptera, Pyralidae) eggs held at 0, 4 and 8°C and for up to 31 days was measured. Parasitism was lowest on eggs held at 8°C and highest on eggs held at 0°C. The highest parasitism, 97.8%, was measured for parasitoids attacking eggs held for 3 days and stored at 0°C. Parasitism of eggs stored at all three temperatures decreased with increasing duration of storage. The number of T. cacoeciae successfully developing and emerging as adults after storage in E. kuehniella eggs held at 0, 4 and 8°C was measured. Parasitoid emergence was >83% from E. kuehniella eggs stored at 8°C for 3 weeks. Storage at 0°C caused a significant decline in parasitoid emergence after 2 weeks (P<0.05). Storage at 0°C for more than 4 weeks reduced fecundity by 50%. T. cacoeciae parasitized the highest number of E. kuehniella eggs 1 day after adult emergence. The oviposition period lasted 6–7 days, although the parasitoids lived up to 13–14 days. Impact of storage time and temperature on parasitism rates by T. cacoeciae stored while in E. kuehniella eggs was measured. As storage time and temperature increased, subsequent parasitism rates of resulting adult T. cacoeciae decreased. Eggs of E. kuehniella can be stored at 0°C for up to 31 days. Trichogramma cacoeciae developing in eggs of E. kuehniella can be stored at 4°C for up to 5 weeks prior to release.     

Trichogramma egg parasitism of Helicoverpa armigera on pigeonpea and sorghum in southern India

Trichogramma spp. (Hymenoptera: Trichogrammatidae) only rarely parasitize eggs of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) on pigeonpea (Cajanus cajan (L.) Millspaugh), while on other plants including sorghum (Sorghum bicolor (L.)) high parasitism levels of this host are found. In this study two strategies designed to increase the parasitoid activity on pigeonpea were tested in the field: intercropping pigeonpea with sorghum and mass-releasing T. chilonis Ishii. Neither strategy led to an increase in parasitism. On pigeonpea, H. armigera oviposited >74.8% of its eggs on calyxes and pods. Parasitism levels in host eggs collected from different plant structures varied significantly with 3.6, 0.3, and 40.7% of eggs on calyxes, pods, and leaves parasitized. Earlier studies have shown that calyxes and pods possess long glandular and non-glandular trichomes, and are covered by sticky trichome exudates which inhibit parasitoid searching behaviour. Parasitism levels between 27.9 and 100% were recorded from host eggs on the intercropped sorghum. Trichogramma chilonis was the dominant parasitoid species. The mean clutch size was 2.03, but up to six parasitoids emerged per egg. Progeny sex ratio (% females) decreased with clutch size, from 63.1% at a clutch size of one to 46.0% at a clutch size of five. Sticky trap catches showed that while the parasitoid population in sorghum increased when H. armigera started ovipositing, the population within pigeonpea did not benefit from either a high parasitoid population in sorghum or a high host egg density on pigeonpea. During one of five seasons studied, however, high parasitism levels (up to 73%) were recorded on pigeonpea. During this season, H. armigera oviposited on pigeonpea plants in the vegetative growth stage and a high proportion of eggs were collected from leaves. Parasitism levels were positively correlated with the percentage of eggs collected from leaves. This study shows that the parasitization efficiency of Trichogramma spp. on pigeonpea depends mainly on the location of the host eggs. This explains why parasitism levels of H. armigera eggs on pigeonpea did not increase when intercropped with sorghum or after mass-releasing T. chilonis.     

Parasitization of potato tuber moth eggs (Lep., Gelechiidae) from irradiated adults by Trichogramma (Hym., Trichogrammatidae) and control of moth population with combined releases of sterile insect and egg parasitoid

Abstract:  Eggs of the potato tuber moth (PTM) Phthorimaea operculella (Zeller) obtained from 150 or 300 Gy-irradiated moths, either inbred or out-crossed to untreated counterparts, were offered as host material to three generalist Trichogramma species; Trichogramma cacoeciae , Trichogramma evanescens , and Trichogramma principium , to determine the ability of these egg parasitoids to develop in F₁ sterile eggs. All eggs laid by the different parental crosses at 150 or 300 Gy, were acceptable and suitable for Trichogramma development. However, in no choice and paired comparison tests, eggs from 300 Gy-irradiated parents were significantly less preferred compared with eggs laid by non-irradiated moths. No intra-specific differences for PTM sterile eggs were recorded among the tested Trichogramma species. The mean number of wasps emerging from each egg types was not affected by an interaction between parental crosses and applied doses. Overall, this study demonstrates that release of irradiated and non-irradiated moths at a 5 : 1 over-flooding ratio and Trichogramma over potatoes in small plexiglass boxes, is complementary and more effective in reducing PTM F₁-emerged progeny than either technique used alone. The results provide baseline data for developing an integrated control approach by using inherited sterility technique in conjunction with Trichogramma for PTM population suppression.