Egg parasitoids of the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), from south-east Queensland

Field surveys of egg parasitoids of the diamondback moth, Plutella xylostella, were conducted at Redlands and Gatton, south-east Queensland. Eggs of P. xylostella were present all year round in both localities, and parasitized eggs were consistently found between late spring and early winter. Percent parasitism in the range 30-75% was recorded on many occasions, although rates less than 10% were more common. The major parasitoids included Trichogrammatoidea bactrae Nagaraja and Trichogramma pretiosum Riley. Laboratory evaluation showed that the T. pretiosum from Gatton has a high capacity to parasitize P. xylostella eggs under suitable conditions. This study represents the first record of egg parasitoids of P. xylostella from Australia.

Cannibalistic feeding of larval Trichogramma carverae parasitoids in moth eggs

Wasps of the genus Trichogramma parasitise the eggs of Lepidoptera. They may deposit one or many eggs in each host. Survival is high at low density but reaches a plateau as density increases. To reveal the mechanism by which excess larvae die we chose a lepidopteran host that has flattened, transparent eggs and used video microscopy to record novel feeding behaviours and interactions of larval Trichogramma carverae (Oatman and Pinto) at different densities. Single larvae show a rapid food ingestion phase, followed by a period of extensive saliva release. Ultimately the host egg is completely consumed. The larva then extracts excess moisture from the egg, providing a dry environment for pupation. When multiple larvae are present, the initial scramble for food results in the larvae consuming all of the egg contents early in development. All larvae survive if there is sufficient food for all to reach a threshold developmental stage. If not, physical proximity results in attack and consumption of others, continuing until the surviving larvae reach the threshold stage beyond which attacks seem to be no longer effective. The number of larvae remaining at the end of rapid ingestion dictates how many will survive to emerge as adults.

Possible incompatibility between transgenic cottons and parasitoids

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 Hubner, 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.

Lack of intraspecific biological variation between two geographical populations of Oomyzus sokolowskii (Hymenoptera: Eulophidae), a gregarious larval–pupal parasitioid of Plutella xylostella (Lepidoptera: Plutellidae)

The chalcid, Oomyzus sokolowskii Kurdjumov has been recorded in many parts of the world as a major larval-pupal, gregarious endoparasitoid of the diamondback moth, Plutella xylostella (Linnaeus), a serious pest of brassica vegetable crops worldwide. This study investigated intraspecific variation between two populations of O. sokiolowskii, one from Cape Verde Islands, West Africa and the other from Hangzhou, China. In all crosses and backcrosses between the two geographical populations, the numbers of progeny and sex ratio of progeny were similar to those obtained within each of the populations, demonstrating complete reproductive compatibility between the two populations. The two populations showed similar responses to temperature with respect to development time and survival of immature stages. Observations on the interactions between the two O. sokolowskii populations and Cotesia plutellae (Kurdjumov), another major parasitoid of P. xylostella, showed that neither population could achieve successful parasitism of P. xylostella larvae already parasitized by C. plutellac. However, both O. sokolowskii populations could achieve hyperparasitism by ovipositing into a mid-late stage larva of C. plutellae developing inside the primary host. Contrary to earlier reports, no evidence of intraspecific variations in ability to hyperparasitize between these two populations of O. sokolowskii was found.

Is the potential of Coccidoxenoides perminutus, a mealybug parasitoid, limited by climatic or nutritional factors?

The encyrtid Coccidoxenoides perminutus is a widely distributed parasitoid of citrus mealybug (Planococcus citri). Worldwide, it has been implicated in successful biocontrol in only a few widely separated localities. C perminutus contributes little to control P. citri in field situations in south-east Queensland, Australia, but invades insectary cultures and reduces mealybug populations considerably under these controlled conditions. This discrepancy between poor field performance and good performance under controlled conditions was investigated to establish whether climatic factors inhibit the field performance of this species in the biological control of P. citri. Subsequent laboratory examination of the influence of varied humidities and temperatures on the activity levels and survival of C perminutus revealed a low tolerance for high saturation deficits (i.e., low % RH at high T degreesC) with reduced reproductive output. The influence of different food sources on adult survival and reproduction was also quantified, to establish if the adverse effects of climate could be overcome by supplementing adult diet. Neither honeydew from their mealybug hosts nor nectar from Alphitonia flowers significantly enhanced parasitoid survival. A subsequent test of five nectar species revealed a significant difference in their influence on C. perminutus survival and reproduction, with only Alpinia zerumbet proving to be as suitable as honey. The floral species that proved suitable in the laboratory need to be checked for their attractiveness to C perminutus in the field and for their ability to enhance the survival and reproductive output of parasitoids. This information suggests that the prevailing dry conditions in south-east Queensland citrus-growing areas apparently impede successful biological control of P. citri by C perminutus, but possibilities are available for habitat manipulation (by providing suitable nectar sources for adult parasitoids) to conserve and enhance C perminutus activity in the field. (C) 2004 Elsevier Inc. All rights reserved.

The effect of parasitization by Trichogramma australicum on Helicoverpa armigera host eggs and embryos

Histological investigations of the pathology of Helicoverpa armigera (Hiibner) eggs after attack by the egg parasitoid, Trichogramma australicum (Girault), indicate that the developing embryo is immediately killed by envenomation. Soon afterward the histological staining characteristics of parasitized host embryos change and the embryonic germ band dissociates into a mass of individual rounded cells. Hosts attacked by females sterilized by gamma-irradiation showed the same pathological effects as normally parasitized hosts, indicating that host degeneration is due to female venom rather than factors derived from the parasitoid embryo or larva. Cell death also occurred in older host embryos although tissue breakdown was delayed. These findings have allowed us to determine not just that the host dies but what happens to the cells and tissues, i.e., their physical appearance, the time course of their degeneration, and that the process is retarded in older hosts. These processes can possibly be emulated in artificial diets. (C) 2003 Elsevier Inc. All rights reserved.

Role of Diadegma semiclausum (Hymenoptera: Ichneumonidae) in controlling Plutella xylostella (Lepidoptera: Plutellidae): Cage exclusion experiments and direct observation

We evaluated the role of the larval parasitoid, Diadegma semiclausum Hellen (Hymenoptera: Ichneumonidae), in controlling Plutella xylostella (L.) (Lepidoptera: Plutellidae) by cage exclusion experiments and direct field observation during the winter season in southern Queensland, Australia. The cage exclusion experiment involved uncaged, open cage and closed cage treatments. A higher percentage (54-83%) of P. xylostella larvae on sentinel plants were lost in the uncaged treatment than the closed (4-9%) or open cage treatments (11-29%). Of the larvae that remained in the uncaged treatment, 72-94% were parasitized by D. semiclausum , much higher than that in the open cage treatment (8-37% in first trial, and 38-63% in second trial). Direct observations showed a significant aggregation response of the field D. semiclausum populations to high host density plants in an experimental plot and to high host density plots that were artificially set-up near to the parasitoid source fields. The degree of aggregation varied in response to habitat quality of the parasitoid source field and scales of the manipulated host patches. As a result, density-dependence in the pattern of parasitism may depend on the relative degree of aggregation of the parasitoid population at a particular scale. A high degree of aggregation seems to be necessary to generate density-dependent parasitism by D. semiclausum . Integration of the cage exclusion experiment and direct observation demonstrated the active and dominant role of this parasitoid in controlling P. xylostella in the winter season. A biologically based IPM strategy, which incorporates the use of D. semiclausum with Bt, is suggested for the management of P. xylostella in seasons or regions with a mild temperature.

Evaluation of the impact of natural enemies on Plutella xylostella L. (Lepidoptera: Yponomeutidae) populations on commercial Brassica farms

1 Accurate assessment of the impact of natural enemies on pest populations is fundamental to the design of robust integrated pest management programmes. In most situations, diseases, predators and parasitoids act contemporaneously on insect pest populations and the impact of individual natural enemies, or specific groups of natural enemies, is difficult to interpret. These problems are exacerbated in agro-ecosystems that are frequently disrupted by the application of insecticides. 2 A combination of life-table and natural enemy exclusion techniques was utilized to develop a method for the assessment of the impact of endemic natural enemies on Plutella xylostella populations on commercial Brassica farms. 3 At two of the experimental sites, natural enemies had no impact on P. xylostella survival, at two other sites, natural enemy impact was low but, at a fifth site, natural enemies drastically reduced the P. xylostella population. 4 The calculation of marginal death rates and associated k-values allowed the comparison of mortality factors between experimental sites, and indicated that larval disappearance was consistently the most important mortality factor, followed by egg disappearance, larval parasitism and pupal parasitism. The appropriateness of the methods and assumptions made to calculate the marginal death rates are discussed. 5 The technique represents a robust and easily repeatable method for the analysis of the activity of natural enemies of P. xylostella, which could be adapted for the study of other phytophagous pests.

Experimental analysis of the influence of pest management practice on the efficacy of an endemic arthropod natural enemy complex of the diamondback moth.

Maximizing the contribution of endemic natural enemies to integrated pest management (IPM) programs requires a detailed knowledge of their interactions with the target pest. This experimental field study evaluated the impact of the endemic natural enemy complex of Plutella xylostella (L.) (Lepidoptera: Yponomeutidae) on pest populations in commercial cabbage crops in southeastern Queensland, Australia. Management data were used to score pest management practices at experimental sites on independent Brassica farms practicing a range of pest management strategies, and mechanical methods of natural enemy exclusion were used to assess the impact of natural enemies on introduced cohorts of P. xylostella at each site. Natural enemy impact was greatest at sites adopting IPM and least at sites practicing conventional pest management strategies. At IPM sites, the contribution of natural enemies to P. xylostella mortality permitted the cultivation of marketable crops with no yield loss but with a substantial reduction in insecticide inputs. Three species of larval parasitoids (Diadegma semiclausum Hellen [Hymenoptera: Ichneumonidae], Apanteles ippeus Nixon [Hymenoptera: Braconidae], and Oomyzus sokolowskii Kurdjumov [Hymenoptera: Eulophidae]) and one species of pupal parasitoid Diadromus collaris Gravenhorst (Hymenoptera: Ichneumonidae) attacked immature P. xylostella. The most abundant groups of predatory arthropods caught in pitfall traps were Araneae (Lycosidae) > Coleoptera (Carabidae, Coccinelidae, Staphylinidae) > Neuroptera (Chrysopidae) > Formicidae, whereas on crop foliage Araneae (Clubionidae, Oxyopidae) > Coleoptera (Coccinelidae) > Neuroptera (Chrysopidae) were most common. The abundance and diversity of natural enemies was greatest at sites that adopted IPM, correlating greater P. xylostella mortality at these sites. The efficacy of the natural enemy complex to pest mortality under different pest management regimes and appropriate strategies to optimize this important natural resource are discussed.

Parasitization of Manduca sexta larvae by the parasitoid wasp Cotesia congregata induces an impaired host immune response

During oviposition, the parasitoid wasp Cotesia congregata injects polydnavirus, venom, and parasitoid eggs into larvae of its lepidopteran host.. the tobacco hornworm, Manduca sexta. Polydnaviruses (PDVs) suppress the immune system of the host and allow the juvenile parasitoids to develop without being encapsulated by host hemocytes mobilized by the immune system. Previous work identified a gene in the Cotesia rubecula PDV (CrV1) that is responsible for depolymerization of actin in hemocytes of the host Pieris rapae during a narrow temporal window from 4 to 8 h post-parasitization. Its expression appears temporally correlated with hemocyte dysfunction. After this time, the hemocytes recover, and encapsulation is then inhibited by other mechanism(s). In contrast, in parasitized tobacco hornworm larvae this type of inactivation in hemocytes of parasitized M. sexta larvae leads to irreversible cellular disruption. We have characterized the temporal pattern of expression of the CrV1-homolog from the C. congregata PDV in host fat body and hemocytes using Northern blots, and localized the protein in host hemocytes with polyclonal antibodies to CrV1 protein produced in P. rapae in response to expression of the CrV1 protein. Host hemocytes stained with FITC-labeled phalloidin, which binds to filamentous actin, were used to observe hemocyte disruption in parasitized and virus-injected hosts and a comparison was made to hemocytes of nonparasitized control larvae. At 24 h post-parasitization host hemocytes were significantly altered compared to those of nonparasitized larvae. Hemocytes front newly parasitized hosts displayed blebbing, inhibition of spreading and adhesion, and overall cell disruption. A CrV1-homolog gene product was localized in host hemocytes using polyclonal CrV1 antibodies, suggesting that CrV1-like gene products of C. congregata's bracovirus are responsible for the impaired immune response of the host. (C) 2005 Elsevier Ltd. All rights reserved.

Are insect immune suppressors driving cellular uptake reactions?

Many insect parasitoids that deposit their eggs inside immature stages of other insect species inactivate the cellular host defence to protect the growing embryo from encapsulation. Suppression of encapsulation by polydnavirus-encoded immune-suppressors correlates with specific alterations in hemocytes, mainly cytoskeletal rearrangements and actin-cytoskeleton breakdown. We have previously shown that the Cotesia rubecula polydnavirus gene product CrV1 causes immune suppression when injected into the host hemocoel. CrV1 is taken up by hemocytes although no receptors have been found to bind the protein. Instead CrV1 uptake depends on dimer formation, which is required for interacting with lipophorin, suggesting a CrV1-lipophorin complex internalisation by hemocytes. Since treatment of hemocytes with oligomeric lectins and cytochalasin D can mimic the effects of CrV1, we propose that some dimeric and oligomeric adhesion molecules are able to cross-link receptors on the cell surface and depolymerise actin by leverage-mediated clearance reactions in the hemolymph.

Confirmation of the existence of alloparasitoids in nature - host relationships of an Australian Coccophagus species that parasitizes mealy bugs

Heteronomous hyperparasitoids are parasitic wasps with sex-related host relationships that are unique to a group of genera in the chalcidoid family Aphelinidae. Females are primary parasitoids of various sedentary bugs (mainly, scale insects, mealy bugs, and whiteflies). Males, in contrast, are hyperparasitic, and they frequently develop at the expense of female conspecifics. Alloparasitoids constitute a special category of heteronomous hyperparasitoids, for their males never develop through female conspecifics. The existence of alloparasitic host relationships and the utility of the category 'alloparasitoid' have both been questioned. Here, we present results that confirm the existence of the alloparasitic way of life among heteronomous aphelinids. We investigated an undescribed species of Coccophagus (Hymenoptera: Aphelinidae), an Australian parasitoid that attacks the introduced lantana mealy bug, Phenacoccus parvus Morrison (Homoptera: Pseudococcidae), in Queensland. A year-long field survey regularly returned large numbers of female Coccophagus spec. near gurneyi individuals from P. parvus (total n = 4212), but only few males (n = 11). Males emerged from samples only when the encyrtid parasitoid Anagyrus diversicornis (Howard) (Hymenoptera: Encyrtidae) was present in samples in relatively high numbers. Laboratory oviposition tests confirmed that A. diversicornis is a male host and showed that males do not develop at the expense of conspecific females. Other studies show that males are attracted in numbers to virgin females held in cages above mealy bug-infested Lantana montevidensis (Spreng.) Briq. (Verbenaceae) in the field, demonstrating that they are common in the population as a whole. This confirms that the males need hosts other than conspecific females and that their usual hosts are present outside of the lantana/P. parvus system. The implications of these results for developing a realistic classification of heteronomous host relationships are discussed.

A calreticulin-like protein from endoparasitoid venom fluid is involved in host hemocyte inactivation

During oviposition, most endoparasitoid wasps inject maternal factors into their hosts to interfere with host immune reactions and ensure successful development of their progeny. Since encapsulation is a major cellular defensive response of insects against intruding parasites, parasitoids have developed numerous mechanisms to suppress the host encapsulation capability by interfering with every step in the process, including recognition, adherence and spreading. In previous studies, components of Cotesia rubecula venom were shown to inhibit melanization of host hemolymph by interfering with the prophenoloxidase activation cascade and facilitate expression of polydnavirus genes. Here we report the isolation and characterization of another venom protein with similarity to calreticulin. Results indicate that C rubecula calreticulin (CrCRT) inhibits hemocyte spreading behavior, thus preventing encapsulation of the developing parasitoid. It is possible that the protein might function as an antagonist competing for binding sites with the host hemocyte calreticulin, which mediates early-encapsulation reactions. (c) 2005 Elsevier Ltd. All rights reserved.

Oviposition and reproductive performance of a generalist parasitoid (Trichogramma pretiosum) exposed to host species that differ in their physical characteristics

The response of generalist egg parasitoids to alternative natural hosts that are present simultaneously is not well known. We investigated the behavior of Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) in relation to two field hosts Helicoverpa armigera Hubner and Spodoptera litura Fabricius, in choice and no choice tests. We quantified the effects of natal host species and post-emergence adult age on the oviposition preference of the parasitoids. H. armigera eggs were consistently preferred over S. litura eggs, regardless of the natal host and adult age. When only S. litura eggs were available as hosts, they were parasitized at statistically similar rates to H. armigera eggs (average of 17 +/- 2.7 vs. 13 +/- 3.0, H. armigera to S. litura). The adult lifespan and lifetime fecundity of T. pretiosum were variable but were affected by natal host species and/or host species to which they were exposed. Mean lifespan and fecundity of parasitoids that had developed in H. armigera eggs and were exposed to H. armigera eggs for oviposition were 13.9 +/- 1.8 days and 98.7 +/- 11.0 adult offspring. By contrast, those that developed in S. litura eggs and were exposed to S. litura eggs for oviposition lived for 7 +/- 0.9 days and produced 53.8 +/- 8.0 adult offspring. The ovigeny index (OI) was significantly lower in the parasitoids exposed to H. armigera eggs than in those exposed to S. litura eggs, regardless of the natal host, indicating that H. armigera eggs sustain the adult parasitoids better than S. litura eggs. These results are used to predict parasitoid behavior in the field when both hosts are available. (c) 2006 Elsevier Inc. All rights reserved.