Comparing and contrasting development and reproductive strategies in the pupal hyperparasitoids Lysibia nana and Gelis agilis (Hymenoptera: Ichneumonidae)

In most animals, the optimal phenotype is determined by trade-offs in life-history traits. Here, I compare development and reproductive strategies in two species of solitary secondary hyperparasitoids, Lysibia nana and Gelis agilis, attacking pre-pupae of their primary parasitoid host, Cotesia glomerata. Parasitoid larvae of both species exploit a given amount of host resources with similar efficiency. However, adults exhibit quite different reproductive strategies. Both species are synovigenic, and female wasps emerge with no mature eggs. However, G. agilis must first host-feed to produce eggs, while L. nana does not host-feed but mobilizes internal resources carried over from larval feeding to initiate oogenesis. Further, G. agilis is wingless, produces large eggs, has a long life-span, and generates only small numbers of progeny per day, whereas these traits are reversed in L. nana. Given unlimited hosts, the fecundity curve in L. nana was “front-loaded,” whereas in G. agilis it was depressed and extended over much of adult life. In L. nana (but not G. agilis), wasps provided with honey but no hosts lived significantly longer than wasps provided with both honey and hosts. Differences in the fecundity curves of the two hyperparasitoids are probably based on differing costs of reproduction between them, with the wingless G. agilis much more constrained in finding hosts than the winged L. nana. Importantly, L. nana is known to be a specialist hyperparasitoid of gregarious Cotesia species that pupate in exposed locations on the food plant, whereas Gelis sp. attack and develop in divergent hosts such as parasitoid cocoons, moth pupae and spider egg sacs. Consequently, there is a strong match between brood size in C. glomerata and egg production in L. nana, but a mismatch between these parameters in G. agilis.     

Experimental heatwaves facilitate invasion and alter species interactions and composition in a tropical host-parasitoid community

As mean temperatures increase and heatwaves become more frequent, species are expanding their distributions to colonise new habitats. The resulting novel species interactions will simultaneously shape the temperature-driven reorganization of resident communities. The interactive effects of climate change and climate change-facilitated invasion have rarely been studied in multi-trophic communities, and are likely to differ depending on the nature of the climatic driver (i.e., climate extremes or constant warming). We re-created under laboratory conditions a host-parasitoid community typical of high-elevation rainforest sites in Queensland, Australia, comprising four Drosophila species and two associated parasitoid species. We subjected these communities to an equivalent increase in average temperature in the form of periodic heatwaves or constant warming, in combination with an invasion treatment involving a novel host species from lower-elevation habitats. The two parasitoid species were sensitive to both warming and heatwaves, while the demographic responses of Drosophila species were highly idiosyncratic, reflecting the combined effects of thermal tolerance, parasitism, competition, and facilitation. After multiple generations, our heatwave treatment promoted the establishment of low-elevation species in upland communities. Invasion of the low-elevation species correlated negatively with the abundance of one of the parasitoid species, leading to cascading effects on its hosts and their competitors. Our study, therefore, reveals differing, sometimes contrasting, impacts of extreme temperatures and constant warming on community composition. It also highlights how the scale and direction of climate impacts could be further modified by invading species within a bi-trophic community network.     

Intrinsic competition between primary hyperparasitoids of the solitary endoparasitoid Cotesia rubecula

1. In nature, competitive interactions occur when different species exploit similar niches. Parasitic wasps (parasitoids) often have narrow host ranges and need to cope with competitors that use the same host species for development of their offspring. When larvae of different parasitoid species develop in the same host, this leads to intrinsic and often contest competition. Thus far, most studies on intrinsic competition have focused on primary parasitoids. However, competition among primary hyperparasitoids, parasitic wasps that use primary parasitoids as a host, has been little studied. 2. This study investigated intrinsic competition between two primary hyperparasitoids, the gregarious Baryscapus galactopus and the solitary Mesochorus gemellus, which lay their eggs in primary parasitoid larvae of Cotesia rubecula, while those in turn are developing inside their herbivore host, Pieris rapae. The aims were to identify: (i) which hyperparasitoid is the superior competitor; and (ii) whether oviposition sequence affects the outcome of intrinsic competition. 3. The results show that B. galactopus won 70% of contests when the two hyperparasitoids parasitised the host at the same time, and 90% when B. galactopus oviposited first. When M. gemellus had a 48 h head start, the two hyperparasitoids had an equal chance to win the competition. This suggests that B. galactopus is an intrinsically superior competitor to M. gemellus. Moreover, the outcome of competition is affected by time lags in oviposition events. 4. In contrast to what has been reported for primary parasitoids, we found that a gregarious hyperparasitoid species had a competitive advantage over a solitary species.     

Recognition of heterospecific parasitism: Competition between Aphidiid (Aphidius ervi) and Aphelinid (Aphelinus asychis) parasitoids of Aphids (Hymenoptera: Aphidiidae; Aphelinidae)

The solitary parasitoids Aphidius erviHaliday (Hymenoptera: Aphidiidae) and Aphelinus asychisWalker (Hymenoptera: Aphelinidae) attacked but generally did not oviposit in pea aphids parasitized by the other species. Wasps selectively oviposited in unparasitized hosts when given a choice. Host discrimination depended on the recognition of internal cues. Females of A. asychiseither could not recognize or ignored A. ervi'sexternal host marking pheromone. Under most conditions, A. ervisurvived in superparasitized hosts, killing competing A. asychislarvae by physical attack and possibly physiological suppression. The outcome of larval competition was not affected by oviposition sequence or age difference between larvae; A. asychissurvived only when it had substantially completed larval development before the host was superparasitized by A. ervi.It is suggested that competition for host resources incurs a cost, for the winner in terms of reduced size or increased development time and for the loser in terms of lost progeny and searching time. Consequently, heterospecific host discrimination can be functional. Internal, and probably general, cues enable wasps to recognize and avoid oviposition in hosts already parasitized by an unrelated species.     

Range size and growth temperature influence Eucalyptus species responses to an experimental heatwave

Understanding forest tree responses to climate warming and heatwaves is important for predicting changes in tree species diversity, forest C uptake, and vegetation–climate interactions. Yet, tree species differences in heatwave tolerance and their plasticity to growth temperature remain poorly understood. In this study, populations of four Eucalyptus species, two with large range sizes and two with comparatively small range sizes, were grown under two temperature treatments (cool and warm) before being exposed to an equivalent experimental heatwave. We tested whether the species with large and small range sizes differed in heatwave tolerance, and whether trees grown under warmer temperatures were more tolerant of heatwave conditions than trees grown under cooler temperatures. Visible heatwave damage was more common and severe in the species with small rather than large range sizes. In general, species that showed less tissue damage maintained higher stomatal conductance, lower leaf temperatures, larger increases in isoprene emissions, and less photosynthetic inhibition than species that showed more damage. Species exhibiting more severe visible damage had larger increases in heat shock proteins (HSPs) and respiratory thermotolerance (T_max). Thus, across species, increases in HSPs and T_max were positively correlated, but inversely related to increases in isoprene emissions. Integration of leaf gas‐exchange, isoprene emissions, proteomics, and respiratory thermotolerance measurements provided new insight into mechanisms underlying variability in tree species heatwave tolerance. Importantly, warm‐grown seedlings were, surprisingly, more susceptible to heatwave damage than cool‐grown seedlings, which could be associated with reduced enzyme concentrations in leaves. We conclude that species with restricted range sizes, along with trees growing under climate warming, may be more vulnerable to heatwaves of the future.     

Trees tolerate an extreme heatwave via sustained transpirational cooling and increased leaf thermal tolerance

Heatwaves are likely to increase in frequency and intensity with climate change, which may impair tree function and forest C uptake. However, we have little information regarding the impact of extreme heatwaves on the physiological performance of large trees in the field. Here, we grew Eucalyptus parramattensis trees for 1 year with experimental warming (+3°C) in a field setting, until they were greater than 6 m tall. We withheld irrigation for 1 month to dry the surface soils and then implemented an extreme heatwave treatment of 4 consecutive days with air temperatures exceeding 43°C, while monitoring whole‐canopy exchange of CO₂ and H₂O, leaf temperatures, leaf thermal tolerance, and leaf and branch hydraulic status. The heatwave reduced midday canopy photosynthesis to near zero but transpiration persisted, maintaining canopy cooling. A standard photosynthetic model was unable to capture the observed decoupling between photosynthesis and transpiration at high temperatures, suggesting that climate models may underestimate a moderating feedback of vegetation on heatwave intensity. The heatwave also triggered a rapid increase in leaf thermal tolerance, such that leaf temperatures observed during the heatwave were maintained within the thermal limits of leaf function. All responses were equivalent for trees with a prior history of ambient and warmed (+3°C) temperatures, indicating that climate warming conferred no added tolerance of heatwaves expected in the future. This coordinated physiological response utilizing latent cooling and adjustment of thermal thresholds has implications for tree tolerance of future climate extremes as well as model predictions of future heatwave intensity at landscape and global scales.     

Small-scale spatial resource partitioning in a hyperparasitoid community

Plant-herbivore-natural enemy associations underpin ecological communities, and such interactions may go up to four (or even more) trophic levels. Here, over the course of a growing season, we compared the diversity of secondary hyperparasitoids associated with a common host, Cotesia glomerata, a specialized larval endoparasitoid of cabbage butterfly caterpillars that in turn feed on brassicaceous plants. Cocoon clusters of C. glomerata were pinned to ~30 Brassica nigra plants by pinning them either to branches in the canopy (~1.5 m high) or to the base of the stem near the ground. The cocoons were collected a week later and reared to determine which hyperparasitoid species emerged from them. This was done in four consecutive months (June–September). Cocoons placed in the canopy were primarily attacked by specialized winged hyperparasitoids (Lysibia nana, Acrolyta nens), whereas cocoons on the ground were attacked by both winged and generalist wingless hyperparasitoids (Gelis acarorum, G. agilis), although this changed with season. There was much more temporal variation in the diversity and number of species attacking cocoons in the canopy than on the ground; the abundance of L. nana and A. nens varied from month to month, whereas P. semotus was only prevalent in August. By contrast, G. acarorum was abundant in all of the samples placed near the ground. Our results show that hyperparasitoids partition host resources at remarkably small vertical spatial scales. We argue that spatial differences in the distribution of natural enemies can contribute to the diversity patterns observed in the field.     

Intrinsic competition between two secondary hyperparasitoids results in temporal trophic switch

Interspecific competition amongst parasitoids is important in shaping the evolution of life‐history strategies in these insects as well as community structure. Competition for hosts may occur between adult female parasitoids (‘extrinsic’ competition) or their progeny (‘intrinsic’ competition). Here, we examined intrinsic competition between two solitary secondary hyperparasitoids, Lysibia nana and Gelis agilis in cocoons of a primary parasitoid, Cotesia glomerata. Each species was allowed to sting hosts previously parasitized by the other at 24 h time intervals over the course of 144 h (6 days). When hosts were attacked simultaneously, neither species was dominant although the species to attack first won most encounters when it had a 24–48 h head start. However, after this time there was dramatic shift in the outcome with G. agilis dominating in all hosts > 72‐h old, regardless of which species had parasitized C. glomerata first. G. agilis larvae, which initially had competed with L. nana for control of C. glomerata resources, began attacking the larvae of L. nana, whereas L. nana rejected hosts with older G. agilis larvae or pupae. Effects of multiparasitism also affected the development time and adult mass of the winning parasitoid. Our results reveal a shift in the trophic status of G. agilis from C. glomerata (in younger hosts) to L. nana (in older hosts), the first time such a phenomenon has been reported in parasitoids.     

Honeydew composition and its effect on life-history parameters of hyperparasitoids

1. Diets that maximise life span often differ from diets that maximise reproduction. Animals have therefore evolved advanced foraging strategies to acquire optimal nutrition and maximise their fitness. The free-living adult females of parasitoid wasps (Hymenoptera) need to balance their search for hosts to reproduce and for carbohydrate resources to feed. 2. Honeydew, excreted by phloem-feeding insects, presents a widely available carbohydrate source in nature that can benefit natural enemies of honeydew-producing insects. However, the effects of variation in honeydew on organisms in the fourth trophic level, such as hyperparasitoids, are not yet understood. 3. This study examined how five different honeydew types influence longevity and fecundity of four hyperparasitoid taxa. Asaphes spp. (Pteromalidae) and Dendrocerus spp. (Megaspilidae) are secondary parasitoids of aphid parasitoids and are thus associated with honeydew-producing insects. Gelis agilis and Acrolyta nens (both Ichneumonidae) are secondary parasitoids of species that do not use honeydew-producing hosts. 4. Most honeydew types had a positive or neutral effect on life span and fecundity of hyperparasitoids compared with controls without honeydew, although negative effects were also found for both aphid hyperparasitoids. Honeydew produced by aphids feeding on sweet pepper plants was most beneficial for all hyperparasitoid taxa, which can partially be explained by the high amount of honeydew, but also by the composition of dietary sugars in these honeydew types. 5. The findings of this study underline the value of aphid honeydew as a carbohydrate resource for fourth-trophic-level organisms, not only those associated with honeydew-producing insects but also ‘interlopers’ without such a natural association.     

Effect of host‐cocoon mass on adult size in the secondary hyperparasitoid wasp,Pteromalus semotus (Hymenoptera: Pteromalidae)

Abstract Parasitoids have long proven to be model organisms in studying resource‐related constraints on immature development. Here we examine the relationship between host cocoon (= pupal) size in the gregarious endoparasitoid wasp, Cotesia glomerata, and development time and adult size in the solitary idiobiont hyperparasitoid, Pteromalus semotus. Little is known about the biology or ecology of this ecto‐hyperparasitoid species, although it is one of the major secondary hyperparasitoids of C. glomerata. The size of the adult wasp covaried with the size of the host cocoon at parasitism. Moreover, female wasps were larger than male wasps for a given cocoon size. Adult wasps have remarkably long life‐spans, 3 months on average. Longevity did not significantly differ with sex. We also examined how larvae of P. semotus exclude other potential competitors. P. semotus is protandrous, with females taking significantly longer to complete their development than males. In experiments where several eggs of P. semotus were placed on individual pupae of C. glomerata, newly hatched hyperparasitoid larvae moved rapidly over the surface of the host and destroyed the eggs of any conspecifics by biting them before they would initiate feeding on host tissues. Our results are discussed in relation to those with other studies with solitary ichneumonid idiobiont hyperparasitoids of C. glomerata.     

Host preference and offspring performance are linked in three congeneric hyperparasitoid species

1. The optimisation theory predicts that insect mothers should oviposit on resources on which they attain the highest exclusive fitness. The development of parasitoid wasps is dependent on limited host resources that are often not much larger than the adult parasitoid. 2. In the present study preference and development in three congeneric species of secondary hyperparasitoids attacking cocoons of two congeneric primary parasitoids that differ significantly in size were compared. Gelis agilis (Fabricius) and G. acarorum (L.) are wingless hyperparasitoids that forage in grassy habitats, whereas G. areator (Panzer) is fully winged and forages higher in the canopy of forbs. 3. The three species were reared on cocoons containing pupae of a small gregarious endoparasitoid, Cotesia glomerata (L.), and a larger solitary species, C. rubecula (Marshall), both of which develop in the caterpillars of pierid butterflies. 4. Adult mass was correlated with initial cocoon mass in all three species, whereas development time was unaffected. Wasps were larger when developing in C. rubecula. However, for a given host mass, wasps were larger when developing on the smaller host, C. glomerata. This suggests that there is a physiological limit to hyperparasitoid size that was exceeded when C. rubecula served as host. 5. All three hyperparasitoids strongly preferred to attack cocoons of the larger species, C. rubecula, often avoiding cocoons of C. glomerata entirely. 6. Preference and performance are correlated in the three Gelis species. However, owing to variation in the distribution and thus abundance of their hosts, it is argued that cumulative fitness may be still higher in the smaller host species.     

Sex allocation of three solitary ectoparasitic wasp species on bean weevil larvae: Sex ratio change with host quality and local mate competition

Sex ratio manipulation by ovipositing females was surveyed in 3 solitary ectoparastic wasp species,Dinarmus basalis (Pteromalidae),Anisopteromalus calanrae (Pteromalidae), andHeterospilus prosopidis (Braconidae), that parasitize azuki bean weevil (Callosobruchus chinensis (L) (Coleoptera: Buruchidae)) larvae within azuki beans (Vigna angularis). Variables were local mate competition (LMC) and host quality (HQ). We used host age as a measure of host quality (from 9-to 16-day-old hosts), changed the number of ovipositing females to control the level of local mate competition (1 female and 10 females), and examined oviposition patterns of the wasps. The offspring sex ratios (proportion of females) of the 3 wasp species respond qualitatively same to HQ and LMC. The common qualitative tendency among the 3 species is an increase of sex ratios increase with host age. In the process of changing the sex ratio (9–13-day-old) 3 wasp species respond only to HQ. In the hosts that end development in size (14–16-day-old) wasps respond to LMC. The response of sex ratio change to LMC in the old host ageclasses are different among the 3 species. In the situation that there exists LMC (10 females) sex ratios are the same among the 3 wasps. However, the sex ratios in no LMC (single female) are heterogeneous among the 3 wasps.     

The effect of host developmental stage at parasitism on sex‐related size differentiation in a larval endoparasitoid

• 1 For their larval development, parasitoids depend on the quality and quantity of resources provided by a single host. Therefore, a close relationship is predicted between the size of the host at parasitism and the size of the emerging adult wasp. This relationship is less clear for koinobiont than for idiobiont parasitoids.
• 2 As size differentiation in host species exhibiting sexual size dimorphism (SSD) is likely to occur already during larval development, in koinobiont larval endoparasitoids the size of the emerging adult may also be constrained based on the sex of the host caterpillar.
• 3 Sex‐specific growth trajectories were compared in unparasitised Plutella xylostella caterpillars and in second and fourth instar hosts that were parasitised by the solitary larval koinobiont endoparasitoid Diadegma semiclausum. Both species exhibit SSD, where females are significantly larger than males.
• 4 Healthy female P. xylostella caterpillars developed significantly faster than their male conspecifics. Host regulation induced by D. semiclausum parasitism depended on the instar attacked. Parasitism in second‐instar caterpillars reduced growth compared to healthy unparasitised caterpillars, whereas parasitism in fourth‐instar caterpillars arrested development. The reduction in growth was most pronounced in hosts producing male D. semiclausum.
• 5 Parasitism itself had the largest impact on host growth. SSD in the parasitoid is mainly the result of differences in growth rate of the parasitoid–host complex producing male and female wasps and differences in exploitation of the host resources. Female wasps converted host biomass more efficiently into adult biomass than males.

     

Fatty acid composition remains stable across trophic levels in a gall wasp community

Acquiring sufficient nutrients is particularly important for insects that are unable to synthesize certain nutrient types de novo, as is the case for numerous parasitoid species that do not synthesize lipids. The lipid reserves of parasitoids are acquired from a single host during larval development. This imposes constraints on the quantity and quality of available lipids. In the present study, the lipid dynamics throughout the trophic cascade are investigated by measuring lipogenic ability, modifications in fatty acid composition and host exploitation efficiency in species at different trophic positions within the community of parasitoids associated with the gall wasp Diplolepis rosae L. (Hymenoptera: Cynipidae). The results obtained show that lipid levels remain stable or decline after feeding in all species, indicating that none of the wasps synthesize lipids. Fatty acid composition is highly similar between the gall wasp, parasitoid and hyperparasitoid species, with the exception of the parasitoid Orthopelma mediator Thunberg (Hymenoptera: Ichneumonidae). The divergence of fatty acid composition in O. mediator suggests that this species is able to modify its fatty acid composition after the consumption of host lipids. The efficiency of exploitation of host resource, in terms of dry body mass acquired, varies among the species (41–70%), although it is high overall compared with the efficiencies reported in other animals. Hence, for parasitoid wasps that lack lipid synthesis capabilities, the efficiency of host exploitation is high and fatty acids are consumed directly from the host without modification, leading to stable fatty acid compositions throughout the trophic cascade.     

菜蛾啮小蜂的生物学及温度对其 种群增长的影响

菜蛾啮小蜂Oomyzus sokolowskii (Kurdjumov) 是小菜蛾Plutella xylostella L. 的一种主要寄生天敌。观察表明,该蜂喜产卵于小菜蛾各龄幼虫,也可产卵于预蛹,进行幼虫至蛹期的跨期聚寄生。每头寄主蛹出蜂多为5～10头,平均7.8头,其中雌蜂占85%～90%。该蜂也可产卵于在小菜蛾幼虫体内寄生的菜蛾绒茧蜂高龄幼虫,故又是小菜蛾的兼性重寄生蜂。在杭州,每年该蜂在田间的活动期为4～10月,10月中下旬陆续以老熟幼虫或预蛹进入休眠越冬,第二年4月陆续羽化并开始产卵寄生。该蜂发育、存活和繁殖的适温范围为20～30℃,低于20℃或高于30℃对其存活不利,但在适温下发育羽化的雌蜂,短时间内在32～35℃高温下仍可大量产卵寄生。在20℃、25℃和30℃下,平均每雌一生可寄生小菜蛾幼虫数分别为3.1、13.2和6.8头,产子蜂数分别为20.5、92.1和504头,内禀增长率分别为0.082、0.240和0.263(雌/雌·天)。     

Convergent development of a parasitoid wasp on three host species with differing mass and growth potential

Koinobiont parasitoids develop in hosts that continue feeding and growing during the course of parasitism. Here, we compared development of a solitary koinobiont endoparasitoid, Meteorus pulchricornis Westmael (Hymenoptera: Braconidae), in second (L2) and fourth (L4) instars of three host species that are closely related (Lepidoptera: Noctuidae) but which exhibit large variation in growth potential. Two hosts, Mamestra brassicae L. and Spodoptera littoralis Boisduval, may reach 1 g or more when the caterpillars are fully mature, whereas Spodoptera exigua Hübner is much smaller with mature caterpillars rarely exceeding 200 mg. Parasitoid survival (to pupation) in the two host instars was much higher on the larger hosts than on S. exigua. However, other fitness correlates in M. pulchricornis were very similar in the three host species. Development time was fairly uniform in L2 and L4 hosts of the three host species, whereas wasps were larger in L4 than in L2 hosts. However, M. pulchricornis developmentally arrested each of the hosts differently. The mass of dying L2 and L4 hosts after parasitoid larval egression (i.e., when they emerge from the dying caterpillar) varied significantly, with S. littoralis being by far the largest and S. exigua the smallest. These results reveal that M. pulchricornis is able to adjust its own development in response to species‐specific differences in host resources.     

Differing Host Exploitation Efficiencies in Two Hyperparasitoids: When is a ��Match Made in Heaven��?

Host exploitation behavior in two hyperparasitoids, Lysibia nana and Gelis agilis, was compared in single cocoon clusters of their primary parasitoid host, Cotesia glomerata. L. nana reproduces sexually, is fully winged, does not host-feed and matures eggs quite rapidly after eclosion, whereas G. agilis possesses opposite traits. Cohorts of individual hyperparasitoid females of differing age and physiological state were given access to single cocoon clusters of C. glomerata that also varied in age. These results reveal that the reproductive biology of L. nana is well matched to exploit cocoon broods in C. glomerata, suggesting strong a co-evolutionary history with this host. By contrast, G. agilis is much less efficient at exploiting host cocoons and is probably a generalist species that attacks other hosts in nature.     

Postemergence Experience Affects Sex Ratio Allocation in a Gregarious Insect Parasitoid

We tested the hypotheses that postemergence experience with plants (“early adult learning”) modifies sex ratio and clutch size allocations of Cotesia congregata (Say) (Hymenoptera: Braconidae), a gregarious larval endoparasitoid of Manduca sexta L. (Lepidoptera: Sphingidae). Emerging wasps were exposed for 2–3 h to (a) one of two host plants (tomato or tobacco) or no plant, and (b) one of two novel plants (arugula or parsley) or no plant. Each female was permitted a single oviposition in a host offered with one of the two plant species 24 h later. Hosts were reared on laboratory diet before and after parasitization. Wasps exposed to either host plant allocated proportionately more females to hosts offered with the plant species experienced at emergence than wasps with the alternate species, but clutch sizes did not differ. Irrespective of plant species, wasps exposed to novel plants allocated proportionately more females to hosts than wasps without plant experience, and larger clutches to hosts offered with parsley than with arugula. Differential responses to host and novel plants suggest inherent recognition of host foodplants by C. congregata. Results demonstrate a direct effect of learning on reproductive potential.     

Competitive interactions between parasitoid larvae and the evolution of gregarious development

We report experiments using two closely related species of alysiine braconids directed at understanding how gregarious development evolved in one subfamily of parasitoid wasps. Theoretical models predict that once siblicide between parasitoid wasps has evolved, it can only be lost under stringent conditions, making the transition from solitary to gregarious development exiguous. Phylogenetic studies indicate, however, that gregariousness has independently arisen on numerous occasions. New theoretical models have demonstrated that if gregarious development involves reductions in larval mobility, rather than a lack of fighting ability (as in the older models), the evolution of gregariousness is much more likely. We tested the predictions of the older tolerance models (gregariousness based on non-fighting larval phenotypes) and the reduced mobility models (gregariousness based on non-searching larval phenotypes) by observing larval movement and the outcome of interspecific competition between Aphaereta genevensis (solitary) and A. pallipes (gregarious) under multiparasitism. Differences in larval mobility matched the prediction of the reduced mobility model of gregarious development, with the solitary A. genevensis having larvae that are much more mobile. The proportion of hosts producing the solitary species significantly declined after subsequent exposure to females of the gregarious species. This contradicts the prediction of the older models (fighting vs non-fighting phenotypes), under which any competitive interactions between solitary and gregarious larvae will result in a highly asymmetrical outcome, as the solitary species should be competitively superior. The observed outcome of interspecific competition offers evidence, with respect to this subfamily, in favour of the new models (searching vs non-searching phenotypes).