Host Pupae Research Articles

Temporal variability in host availability alters the outcome of competition between two parasitoid species.

Variability in the availability of resources through time is a common attribute in trophic interactions, but its effects on the fitness of different consumer species and on interspecific competition between them are not clearly understood. To investigate this, we allowed two parasitoid species, Trichopria drosophilae and Pachycrepoideus vindemiae, to exploit Drosophila host pupae under different temporal variability treatments, either on their own or simultaneously. When tested individually (in the absence of interspecific competition), both parasitoid species had lower fitness when hosts were exposed for a short duration at high density than when exposed for a long duration at low density. When both parasitoid species exploited hosts simultaneously, interspecific competition significantly decreased the number of offspring for both parasitoid species. The outcome of this interspecific competition depended on host temporal variability, with T. drosophilae or P. vindemiae dominating in short and long host exposure treatments, respectively. These results can be explained by the combination of host availability and egg load of female adult parasitoids. When abundant hosts are provided for a short period, the ample mature eggs of the proovigenic T. drosophilae enable them to exploit hosts more efficiently than P. vindemiae, which is synovigenic. However, P. vindemiae is an intrinsically superior competitor and dominates when multiparasitism occurs. Multiparasitism is more frequent when hosts are at low levels relative to the egg load of the parasitoids. Our results clearly demonstrate that resource temporal availability can alter the outcome of competition between consumers with different reproductive traits.

Long-term suitability of an alternative host for rearing the sugarcane stalk borer parasitoid Tetrastichus howardi.

The continuous utilisation of an alternative host may influence parasitoid performance across successive generations due to conditioning in natal hosts. Tetrastichus howardi (Olliff) has successfully been reared using Tenebrio molitor L. pupae as a feasible alternative host. However, the extended rearing of T. howardi on this alternative host may impact the biological features of the parasitoids. Parasitoids were reared using T. molitor pupae for 30 consecutive generations. Quality criteria were assessed during the generations F5, F15, and F30, offering pupae of the target pest, Diatraea saccharalis (Fabr.), and compared with the F0 generation (parasitoids reared in D. saccharalis pupae). Criteria included assessments of parasitism performance, host selection, and wing form variation in the parasitoid wasps. Additionally, we examined the fecundity of T. howardi females that emerged from both hosts, considering their age, egg loading before and after one oviposition, as well as parasitism of sugarcane stalk borer pupae. Rearing T. howardi using pupae of T. molitor did not affect its biological traits or preference for the target pest for 30 generations. After parasitism, the parasitoid left the host pupa inside the stalk, and one oviposition was enough to kill D. saccharalis pupae and obtain viable parasitoid progeny. Female sexual maturation and egg loading occurred 72 and 96 h after parasitoid emergence. Egg-loading recovery after parasitism did not happen within 24 h. T. howardi can be reared for up to 30 generations using alternative hosts without compromising its parasitism performance or egg loading.

Spodoptera cosmioides (Lepidoptera: Noctuidae) as an alternative host for mass rearing the parasitoid Palmistichus elaeisis (Hymenoptera: Eulophidae)

Abstract Different alternative hosts can reduce or even avoid pre-imaginal conditioning of parasitoids in mass production programs. Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae) may be an alternative host for mass rearing the parasitoid Palmistichus elaeisis Delvare & LaSalle (Hymenoptera: Eulophidae). The objective of this study was to evaluate the parasitism, emergence, egg-to-adult period, progeny size, sex ratio, and consumption of host tissue per individual of P. elaeisis produced using S. cosmioides pupae in the laboratory. Newly formed S. cosmioides pupae were weighed, individualized in glass tubes and each one was exposed to six newly emerged P. elaeisis females during 48 h. These pupae were transferred to plastic pots until the emergence of P. elaeisis adults. Palmistichus elaeisis parasitized and emerged from 100.0 % and 87.5 % of the S. cosmioides pupae, respectively. The egg-to-adult period of P. elaeisis was 24.28 ± 0.18 days, with 109 ± 2 individuals emerging per host pupae. Mass consumption of the host was 2.56 ± 0.01 mg per parasitoid emerged. This is the first report of P. elaeisis parasitizing and developing in S. cosmioides pupae. The ease of rearing S. cosmioides with an artificial diet, and the high parasitism, emergence, and progeny size of P. elaeisis support the suitability of this alternative host to mass-rearing this parasitoid.

Functional response and mutual interference in the parasitoid Coptera haywardi (Oglobin) (Hymenoptera: Diapriidae) attacking Anastrepha ludens (Loew) (Diptera: Tephritidae) pupae.

Functional response and mutual interference are important attributes of natural enemies that should be analysed in species with the potential to be used as biological control agents in order to increase the predictive power of the possible benefits and/or consequences of their release in the field. Our main objective was to determine the functional response and mutual interference of Coptera haywardi (Oglobin), a pupal parasitoid of economically important fruit flies (Diptera: Tephritidae). The functional response of C. haywardi on A. ludens pupae corresponded to a type II model, with an attack rate of 0.0134 host pupa/h and a handling time of 1.843 h, which reveals a meticulous selection process of pupal hosts. The effect of mutual interference among foraging females was negatively correlated with increased parasitoid density in the experimental arena, showing a gradual decline in attack rate per individual female. The increase in the number of foraging females also had an impact on the number of oviposition scars per pupa and the number of immature parasitoids per dissected pupa, but not on the percentage of adult emergence or the sex ratio. Our results suggest that C. haywardi could act as a complementary parasitoid in the control of fruit fly pupae, since the random distribution of these pupae in the soil would decrease the possibility of aggregation and mutual interference between foraging females.

Survey on Drosophila suzukii and Ceratitis capitata (Diptera: Drosophilidae, Tephritidae) and Associated Eucoilinae Species (Hymenoptera: Figitidae) in Northwestern Argentina. First Record of Dicerataspis grenadensis and Leptopilina boulardi as Parasitoids of D. suzukii.

The Southeast Asian-native Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), also known as "spotted-wing Drosophila," is one of the most globally invasive agricultural species. Although D. suzukii is a pest spread throughout all the Argentinian fruit-growing regions, few information has been published on its impact on local fruit production. Parasitoid species associated with D. suzukii in Argentina belong to Pteromalidae (Chalcidoidea), Diapriidae (Diaprioidea), both attacking host pupae, and Figitidae (Cynipoidea), which attack host larvae. Nine Eucoilinae (Figitidae) species, belonging to Dicerataspis, Dieucoila, Euxestophaga, Ganaspis, Hexacola, and Leptopilina genera, have been associated with D. suzukii in Argentina. Ceratitis capitata (Wiedemann), commonly known as "medfly," is native to Africa and has a worldwide distribution, covering many tropical, subtropical, and temperate regions. In Argentina, C. capitata has been associated with several native hymenopterous parasitoids belonging to Braconidae (Ichneumonioidea), Eulophidae (Chalcidoidea), Pteromalidae, Diapriidae, and Figitidae families. Only two eucoline species, Ganaspis pelleranoi (Brèthes) and Leptopilina haywardi (Blanchard) have been related to medfly in Argentina. We report new trophic associations between the parasitoids Dicerataspis grenadensis Ashmead and Leptopilina boulardi (Barbotin, Carton and Kelner-Pillault) and D. suzukii, and between the parasitoid Odontosema albinerve Kieffer and C. capitata, after surveys conducted in Tucumán, northwestern Argentina. An annotated checklist and a taxonomic key of Eucoilinae associated with both invasive pests, in Argentina, are also provided.

Virus infections in honeybee colonies naturally surviving ectoparasitic mite vectors

Western honeybee populations, Apis mellifera, in Europe have been known to survive infestations of the ectoparasitic mite, Varroa destructor, by means of natural selection. Proposed mechanisms in literature have been focused on the management of this parasite, however literature remains scare on the differences in viral ecology between colonies that have adapted to V. destructor and those that are consistently treated for it. Samples were collected from both a mite-surviving and a sympatric mite-susceptible honeybee population in Norway. The prevalence and abundances of 10 viruses, vectored by the parasite or not, were investigated in adult host workers and pupae as well as in V. destructor mites. Here we show that the mite-vectored Deformed wing virus (DWV-A) is often lower in both abundance and prevalence in the mite-surviving population in tandem with lower phoretic mite infestations compared to the mite susceptible population. However, the non-mite-vectored Black queen cell virus (BQCV), had both a higher abundance and prevalence in the mite-surviving population compared to the susceptible population. The data therefore suggest that general adaptations to virus infections may be unlikely to explain colony survival. Instead, mechanisms suppressing mite reproduction and therefore the impact seem to be more important.

Side Effects of Single-Transgene or Pyramided Genetically Modified Maize on the Generalist Endoparasitoid Palmistichus elaeisis (Hymenoptera: Eulophidae)

The fall armyworm, Spodoptera frugiperda J.E. Smith (Lepidoptera: Noctuidae), is an agricultural pest native to tropical and subtropical regions of the Americas, with considerable potential for global invasion and seasonal migration. Although genetically modified (GM) plants have shown positive impacts on the economy and the environment as they synthesize proteins that act as natural insecticides and are primarily intended to protect the crops from damage by specific pests, potential effects of Bt toxins on non-target organisms can occur. This experiment aimed to evaluate the potential impacts on the parasitoid Palmistichus elaeisis (Hymenoptera: Eulophidae), using the pupae of Spodoptera frugiperda (Lepidoptera: Noctuidae) as hosts, which were fed with transgenic maize genotypes such as Herculex®, expressing Cry1F protein, and PowerCore®, expressing Cry1F, Cry1A.105, and Cry2Ab2 proteins, or their non-transgenic isohybrid. The experiment was conducted in a completely randomized design with ten replicates. Spodoptera frugiperda larvae were fed ad libitum with transgenic and non-transgenic maize until the pupal stage and then individually exposed to six P. elaeisis females for 78 h. The total number of adults, emerged males, tibia size, cephalic capsule size, and parasitoid body size were not influenced by host feeding. However, the number of emerged females from the Herculex® and PowerCore® treatments was lower than that for the isohybrid treatment. The sex ratio and longevity of parasitoids emerging from PowerCore® were 1.05 and 1.26 times lower, respectively, than that of those from the isohybrid. Furthermore, the number of dead P. elaeisis within the host pupa was 10.56 times higher in the PowerCore® genotype. Combining Bt proteins within pyramided genotypes should cause minimal impacts and promote the conservation and integration of beneficial insects. The results of this study provide helpful information for developing effective and compatible integrated pest management (IPM) strategies.

Fitness and cold tolerance of Spodoptera frugiperda fed on corn and two winter crops

AbstractFall armyworm (FAW) Spodoptera frugiperda is a major migratory and polyphagous pest. Overwintering, migrating and colonizing characters of FAW are closely related to vegetation and host adaptation. In this study, we investigated oviposition preference, feeding preference of FAW, as well as fitness, cold tolerance of FAW larvae fed on spring and summer crop (corn) and two winter crops (cabbage, Brassica campestris and rape, B. napus). The results showed that FAW could complete their life cycle by feeding on cabbage and rape, although fitness was not as good as corn. FAW fed on cabbage had higher survival, female pupal mass and host suitability index than rape while larvae preferred to feed on rape leaves and females preferred to lay eggs on rape plants. In addition, FAW larvae fed on cabbage had the shortest recovery time from chill coma compared to corn and rape, indicating that cold tolerance of FAW larvae was improved by feeding on cabbage. Consequently, cabbage could be an ideal host for FAW which needed to survive and reproduce in winter. These results improve our understanding of host selection and adaption of FAW, as well as lay a foundation for predicting winter breeding areas of FAW.

Understanding the ability of Pachycrepoideus vindemiae (Hymenoptera: Pteromalidae) to find and parasitize buried Diptera pupae in southwestern Colombia

Pachycrepoideus vindemiae is a generalist wasp parasitoid released for biological control of the pupal stage for several species of Tephritoidea and Muscoidea flies that limit production of fruits and poultry commodities, respectively, worldwide. The parasitoid wasp must find buried host pupae to oviposit on them, and several factors may influence this outcome. The objectives of this study were to determine the capacity of first, host larvae to burrow to pupate according to substrate humidity and second, the parasitoid wasp to burrow and parasitize buried host pupae according to different conditions (substrate type, host species and parasitization time exposure). Moreover, comparison of chemical profiles between host’s pupae potentially involved for host location by the parasitoid was realized by GC-FID. Peat humidity significantly affected the burrowing depth of Dasiops inedulis and Anastrepha striata and both species buried significantly deeper in peat with 50% humidity than in dry conditions. The number of emerged parasitoids is not different between the two tested host species. P. vindemiae performance was better in peat than in crop soil. In peat wasps’ parasitism is similar between buried pupae located at 0 mm or 10 mm depth regardless of the time of exposure to the parasitoid. Cuticular chemical profiles showed that the four-host pupa species studied did not share compounds. However, A. striata shared some compounds with D. inedulis and, M. domestica shared some with C. capitata. Common compounds were found in the extracts of parasitized and non-parasitized pupae of C. capitata, D. inedulis and M. domestica.

Early Stages and Notes on the Biology of Melitaea gina Higgins, 1941 (Lepidoptera: Nymphalidae) from Northern Zagros Mts (Iran) with the Special Remarks on Taxonomy.

This article presents characteristics of the habitats of Melitaea gina Higgins, 1941 and describes the behavior of adults under natural conditions, their host plant, egg chorion, caterpillars, and pupa morphology. The morphology of the early stages and the taxonomy of M. gina are discussed.

Identification of fruit fly (Tephritidae) pupae parasitized by hymenoptera parasitoids

AbstractOne of the major problems in orchards is the presence of Tephritidae fly maggots, especially in Mirtaceae species. Various biological control tools include the use of Hymenoptera parasitoids, which naturally control fly populations. However, identification of the host species of parasitoids is challenging, because only the puparium remains after the parasitoid hatches. Therefore, the aim of this work was to determine the host species of Hymenoptera parasitoids, through the puparium. Host species were identified by amplification and sequencing of cytochrome C oxidase subunit I (COI) of mitochondrial DNA. The results of the present study show that COI DNA sequencing was able to reliably identify host pupae to the genus level, allowing the identification of the Anastrepha genus in this study. This suggests that this method can be used to identify agriculturally relevant hosts through its puparium remains, since this genus has already been described as a pest of Campomanesia adamantium. Knowledge of the species that cause damage to crop is extremely important to study the biological and ecological aspects of these pests and their control. In this way, our results suggest that this method can be used to identify economically relevant host pupal species since the genus identified in this study has already been described as a pest of C. adamantium.

Effect of Short-Term High-Temperature Stimuli on the Functional Response of Trichopria drosophilae (Matsumura).

Researchers have previously investigated the role of Trichopria drosophilae as a pupal parasitoid in the biological control of Drosophila suzukii in China. Here, we investigated the ability of T. drosophilae to parasitize D. suzukii pupae at different temperatures. To do this, we evaluated the functional response of T. drosophilae to D. suzukii pupae at different temperatures and investigated the specific effects of density on parasitism. The results show that the parasitic functional response of T. drosophilae under different high-temperature stimuli is Holling type II. After processing at 29 °C, the instantaneous search rate was 1.1611; the theoretical maximum parasitic value was 20.88 at 31 °C. The parasitic efficiency decreased with increasing stimulation temperature, as the host pupa density increased from 5 to 25, and the strongest search effect occurred at 0.87 at 27 °C. The searching effect of T. drosophilae at each temperature fell gradually with an increase in prey density from 5 to 25. At 31 °C, the theoretical parasitic maximum of T. drosophilae reached a maximum of 20.88 pupae. At this temperature, when a pair of T. drosophilae was placed in a pupa density of 50, its actual total number of parasites was 18.60.

Brain and antennal transcriptomes of host ants reveal potential links between behaviour and the functioning of socially parasitic colonies.

Insect social parasites are characterized by exploiting the hosts' social behaviour. Why exactly hosts direct their caring behaviour towards these parasites and their offspring remains largely unstudied. One hypothesis is that hosts do not perceive their social environment as altered and accept the parasitic colony as their own. We used the ant Leptothorax acervorum, host of the dulotic, obligate social parasite Harpagoxenus sublaevis, to shed light on molecular mechanisms underlying behavioural exploitation by contrasting tissue-specific transcriptomes in young host workers. Host pupae were experimentally (re-)introduced into fragments of their original, another conspecific, heterospecific or parasitic colony. Brain and antennal mRNA was extracted and sequenced from adult ants after they had lived in the experimental colony for at least 50 days after eclosion. The resulting transcriptomes of L. acervorum revealed that ants were indeed affected by their social environment. Host brain transcriptomes were altered by the presence of social parasites, suggesting that the parasitic environment influences brain activity, which may be linked to behavioural changes. Transcriptional activity in the antennae changed most with the presence of unrelated individuals, regardless of whether they were conspecifics or parasites. This suggests early priming of odour perception, which was further supported by sensory perception of odour as an enriched function of differentially expressed genes. Furthermore, gene expression in the antennae, but not in the brain corresponded to ant worker behaviour before sampling. Our study demonstrated that the exploitation of social behaviours by brood parasites correlates with transcriptomic alterations in the central and peripheral nervous systems.

Pathogenicity and Virulence of Different Concentrations of Brazilian Isolates of Entomopathogenic Nematodes Against Drosophila suzukii.

The invasive pest Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) was recently recorded in Brazil and constitutes a threat to fruit growing, mainly for small, soft fruits. Recent advances in research on ways of controlling D. suzukii involve the use of entomopathogenic nematodes (EPNs). In this context, the objective of this study was to evaluate the pathogenicity and virulence of four isolates in different concentrations against D. suzukii pupae. The EPN isolates used in trials were Steinernema brazilense IBCBn 06, S. carpocapsae IBCBn 02, Heterorhabditis bacteriophora HB, and H. amazonensis IBCBn 24. Both H. amazonensis IBCBn 24 and H. bacteriophora HB were effective in controlling D. suzukii as they caused a mortality rate of 86.25% and 80.0%, and virulence of 549.75 IJs/pupae and 787.75 IJs/pupae in the concentrations of 1800 IJs/ml and 5400 IJs/ml, respectively. The lowest lethal concentrations (LC50) of juveniles were found in host pupae with 771.63 IJs/ml of H. bacteriophora HB and 1115.49 IJs/ml of H. amazonensis IBCBn 24. Results showed that both EPNs, H. amazonensis IBCBn 24 and H. bacteriophora HB, could be promising eco-friendly biological agents to control D. suzukii.

Host stage affects oviposition-related behaviours, development progression and reproductive output in a native hyperparasitoid of the solenopsis mealybug invading Asian regions

ABSTRACT Native hyperparasitoids may readily expand their host range by including non-native parasitoids that are introduced in classical biological control of pest insects. Here we report our observation on biology of Cheiloneurus nankingensis (Hymenoptera: Encyrtidae), an obligate hyperparasitoid attacking non-native parasitoid Aenasius arizonensis residing in Phenacoccus solenopsis mealybugs that have invaded into Asian regions. We measured durations of antennae-tapping, ovipositor-drilling and ovipositor-probing/ovipositing behaviours by C. nankingensis wasps on parasitised mealybugs with the host at either larval or pupal stage and on the unparasitised mealybug as the control. The ovipositor-probing/ovipositing duration was much longer on a pupal host than on the larval while shortest on the control. We then observed offspring development progression of C. nankingensis at attacking either egg, larva or pupa of its host by daily dissecting hyperparasitised mealybugs. The development was faster in a larval or pupal host than in the egg. Finally, our measurements of the reproductive output showed that C. nankingensis wasps during 14 days after the emergence attacked more parasitised mealybugs and produced more offspring reaching adulthood when ovipositing in the mealybug with a host larva than with a pupa. Our results from this study suggest that C. nankingensis may employ the ovipositor to probe for the host within a mealybug and the host at the larval stage is of high quality for the reproduction.

The son-killer microbe Arsenophonus nasoniae is a widespread associate of the parasitic wasp Nasonia vitripennis in Europe

Heritable microbes that exhibit reproductive parasitism are common in insects. One class of these are the male-killing bacteria, which are found in a broad range of insect hosts. Commonly, our knowledge of the incidence of these microbes is based on one or a few sampling sites, and the degree and causes of spatial variation are unclear. In this paper, we examine the incidence of the son-killer microbe Arsenophonus nasoniae across European populations of its wasp host, Nasonia vitripennis. In preliminary work, we noticed two female N. vitripennis producing highly female biased sex ratios in a field study from the Netherlands and Germany. When tested, the brood from Germany was revealed to be infected with A. nasoniae. We then completed a broad survey in 2012, in which fly pupal hosts of N. vitripennis were collected from vacated birds’ nests from four European populations, N. vitripennis wasps allowed to emerge and then tested for A. nasoniae presence through PCR assay. We then developed a new screening methodology based on direct PCR assays of fly pupae and applied this to ethanol-preserved material collected from great tit (Parus major) nests in Portugal. These data show A. nasoniae is found widely in European N. vitripennis, being present in Germany, the UK, Finland, Switzerland and Portugal. Samples varied in the frequency with which they carry A. nasoniae, from being rare to being present in 50% of the pupae parasitised by N. vitripennis. Direct screening of ethanol-preserved fly pupae was an effective method for revealing both wasp and A. nasoniae infection, and will facilitate sample transport across national boundaries. Future research should examine the causes of variation in frequency, in particular testing the hypothesis that N. vitripennis superparasitism rates drive the variation in A. nasoniae frequency through providing opportunities for infectious transmission.

Mechanism of adult emergence in Xanthopimpla pedator (Fabricius)- a major pupal parasitoid of Tasar silkworm

Aim: The study was conducted to elucidate the mechanism involved in adult emergence behavior of Xanthopimpla pedator a pupal parasitoid of tasar silkworm. Methodology: Behavioral observations were made to study the mechanical approaches used during emergence and SDS-PAGE analysis was carried to know the possibility of proteases in the digestive secretions. Results: The findings of the study suggest that X. pedator uses physical and chemical to achieve safe passage from the host pupae and cocoons. It was observed that during the process of emergence, X. pedator breaks host pupae by means of mandibular action (by biting the pupal shell). Further, it was noticed that to make emergence hole on host cocoon X. pedator found to employ digestive secretion to soften and disintegrate the silken shell, besides it also deploys mandibles to spread out the secretion on intended place of exit hole. SDS-PAGE analysis showed the presence of series of enzymes, possibly involved in cocoon softening. Interpretation: Findings of this study forms the basis that X. pedator, a major pupal parasitoid of tasar silkworm, uses both physical and chemical approaches to achieve safe passage from the host pupae and protective silken cocoon. Key words: Digestive secretion, Mandibular action, Parasitization, Xanthopimpla pedator

Cadmium (Cd) exposure through Hyphantria cunea pupae reduces the parasitic fitness of Chouioia cunea: A potential risk to its biocontrol efficiency

Heavy metal contamination has been regarded as an environmental variable that affects the efficiency of pest biological control, but the parasitic fitness of parasitoids under heavy metal stress is poorly understood. Herein, the effect of Cd exposure through the host pupa of Hyphantria cunea on the parasitic fitness of Chouioia cunea was investigated, and the mechanism by which Cd exposure affects the interaction between H. cunea and C. cunea from the perspective of innate immunity in host insect and the oxidative status in the parasitoid offspring was explored. Our results indicated that Cd can be transferred from the H. cunea pupae to the parasitoid offspring, and the transfer coefficient reflected biological amplification. There were no significant differences in the rates of parasitism success and offspring emergence between the untreated and Cd-treated groups. However, after parasitizing Cd-accumulated pupae, the parasitic fitness of offspring wasps (e.g., the number, individual size and life span) decreased significantly. Under Cd exposure, the cellular and humoral immunity of H. cunea pupae decreased significantly. Compared with the untreated group, the H2O2 content of parasitoid offspring in the Cd-treated group was significantly increased. Cd exposure significantly inhibited superoxide dismutase activity in parasitoid offspring, but the contents of ascorbic acid and glutathione were significantly increased by Cd stress. Taken together, these results indicate that Cd exposure reduces the cyclic utilization efficiency of C. cunea on H. cunea pupae. The oxidative status of parasitoid offspring triggered by Cd exposure could be responsible for the reduced parasitic fitness of C. cunea on Cd-accumulated H. cunea pupae.

Molecular identification of Cerodontha australis (Diptera: Agromyzidae) and its associated pupal endoparasitoids (Hymenoptera)

AbstractThe wheat sheath miner, Cerodontha australis, is widespread and abundant in New Zealand and also occurs in eastern Australia. Adult and larval C. australis feed on cereals and grasses, including the economically important perennial ryegrass, Lolium perenne. There is little literature about C. australis, and much of the early work may relate to other species due to its initial misidentification. Morphology‐based identification can be challenging, and the absence of online barcode sequences from voucher specimens for C. australis has precluded diagnosis using molecular techniques. In this study, two individual adult leafminers collected from L. perenne plants in New Zealand were confirmed as C. australis morphologically. One has been retained as a voucher specimen and its mitochondrial barcode sequence submitted to online databases. Comparison with adult and larval specimens previously sequenced by the authors confirmed they were also C. australis. Molecular identification of 20 hymenopteran pupal endoparasitoids, and a subset of the puparia they emerged from, revealed Trichopria sp. (Diapriidae), Eupelmus messene (Eupelmidae) and three haplotypes (or closely related species) of Eulophidae. Sequences for all wasps were submitted to GenBank. All the puparia were visually identified as C. australis, and a subset that included a host pupa of each identified endoparasitoid species were confirmed as C. australis based on comparison with our barcode sequence. This work will aid future studies on C. australis and has allowed identification of unidentified sequences on public databases formerly submitted by other New Zealand researchers. It also provides new records of association between C. australis and its pupal endoparasitoids. More broadly, our study provides an example of the importance of authoritative morphological identification of specimens alongside molecular identification, and highlights the challenges in identifying species for which this has not been done.

Adaptive Reproductive Strategies of an Ectoparasitoid Sclerodermus guani under the Stress of Its Entomopathogenic Fungus Beauveria bassiana.

Complex interspecific relationships between parasites and their insect hosts involve multiple factors and are affected by their ecological and evolutionary context. A parasitoid Sclerodermus guani (Hymenoptera: Bethylidae) and an entomopathogenic fungus Beauveria bassiana (Hypocreales: Cordycipitaceae) shared the same host in nature, Monochamus alternatus (Coleoptera: Cerambycidae). They often encountered the semi-enclosed microhabitat of the host larvae or pupae. We tested the survival and reproduction of the parasitoid's parent and its offspring fitness under different concentrations of B. bassiana suspension. The results show that S. guani parent females carrying higher concentrations of the pathogen shorten the pre-reproductive time and regulate their own fertility and their offspring's survival and development. This minimal model of the interspecific interactions contains three dimensionless parameters, vulnerability (θ), dilution ratio (δ), and PR, which were used to evaluate the mortality effect of the parasitoid S. guani on its host M. alternatus under the stress of the entomopathogenic fungus B. bassiana. We compared the infection and lethal effect of the fungus B. bassiana with different concentrations to the parasitoid S. guani and the host larvae M. alternatus. At higher concentrations of the pathogen, the parasitoid parent females shorten the pre-reproductive time and regulate their own fertility and their offspring's survival and development. At moderate concentrations of the pathogen, however, the ability of the parasitoid to exploit the host is more flexible and efficient, possibly reflecting the potential interspecific interactions between the two parasites which were able to coexist and communicate with their hosts in ecological contexts (with a high overlap in time and space) and cause interspecific competition and intraguild predation.