Laboratory investigations of trichomycete prevalence, abundance and fecundity in a Smittium-simuliid model

Smittium, the most speciose genus of the "gut fungi" (Zygomycota: Trichomycetes), is found attached to the hindgut cuticle of larval aquatic Diptera. Smittium spp. colonize several host families (e.g., Smittium culisetae in Chironomidae, Culicidae and Simuliidae), but some species appear to be specific to a single host family (e.g., Smittium morbosum Sweeney in Culicidae). The specificity of Smittium spp. within a host family has been difficult to resolve. This research presents evidence that certain Smittium spp. differentially colonize particular species of black fly (Diptera: Simuliidae) hosts as measured by differences in prevalence, abundance and fecundity. Reasons for this differential occurrence and fecundity in hosts are unclear but might include fungal responses to variations in host morphology, physiology, distribution or behavior. Variable fitness of Smittium spp., within a suite of available hosts, could be a factor in the diversity of this fungal group.     

The genus Smittium (Trichomycetes, Harpellales) in the Iberian Peninsula

A study of larval Diptera (Chironomidae, Culicidae and Simuliidae) from Spain has been carried out to fulfill a catalogue of species of the genus Smittium (Harpellales: Legeriomycetaceae) present within these hosts. Among the reported taxa, eight are new species: Smittium brevisporum, S. bulbosporo-phorus, S. gracilis, S. hecatei, S. heterosporum, S. inex-pectans, S. prostratum and S. pseudodimorphum. We also report six previously described species, which are new for the Iberian Peninsula: S. alpinum, S. dipterorum, S. megazygosporum, S. pusillum, S. typhellum and S. fecundum. Three other species (S. simulii, S. culicis and S. culisetae) previously were reported from Spain. In two of them (S. fecundum and S. culicis), we describe for the first time the presence of zygospores. Some of the included species have been artificially cultured as well as ultrastructurally studied using scanning electron microscopy (SEM), with the purpose of observing the surface of both trichospores and the trichospore collar and the morphology of the trichospore appendage.     

Trichomycete insect symbionts in Great Smoky Mountains National Park and vicinity

Collections of trichomycete symbionts of larval aquatic insects in Great Smoky Mountains National Park and vicinity in the southern Appalachian region of the USA resulted in finding many taxa of Harpellales, including an unusual new monotypic genus, Barbatospora ambicaudata in Simuliidae, and five new species in Thaumaleidae or Chironomidae, Harpellomyces montanus, Smittium lentaquaticum, Sm. minutisporum, Stachylina gravicaudata and St. stenospora. In addition a new species of Amoebidium (Amoebidiales), A. appalachense, attached to the anal tubules of bloodworms (Chironomidae) is described. Axenic cultures of three of the new taxa were obtained, plus Sm. culisetae. Fourteen identified species representing 13 genera of previously known Harpellales are recorded from Plecoptera, Ephemeroptera and Diptera, as well as a new Dipteran host record for an unidentified harpellid that was found in a Blephariceridae. Also identified were Paramoebidium corpulentum and many undetermined species of Paramoebidium (Amoebidiales) from four orders of aquatic insect larvae. The occurrence of an Enterobryus species in Diplopoda and another Eccrinales from an aquatic beetle is noted. Amoebidiales,     

Offspring production and self-superparasitism in the solitary ectoparasitoid Spalangia cameroni (Hymenoptera: Pteromalidae) in relation to host abundance

Parasitoid fitness strongly depends on the availability and quality of hosts, which provide all resources required for larval development. Several factors, such as host size and previous parasitation, may affect host quality. Because self-superparasitism induces competition among a female's offspring, it should only occur if there is an imperfect recognition of self-parasitized hosts or if there is a fitness advantage to self-superparasitism. Against this background, we investigated self-superparasitism and offspring production in Spalangia cameroni (Hymenoptera: Pteromalidae) in relation to the abundance of a novel host, Ceratitis capitata (Diptera: Tephritidae). Individual pairs of parasitoids were provided with either two (low host abundance) or ten (high host abundance) pupae per day. Under high host abundance, lifetime fecundity (number of eggs laid), offspring number, number of pupae parasitized and hosts killed were greater than under low host abundance, whereas the number of eggs per host was lower; and the proportion of hosts that did not produce offspring tended to be lower. The latter suggests the occurrence of ovicide, when hosts are scarce due to an at least imperfect recognition of previously self-parasitized hosts. Offspring production per parasitized pupa was higher when hosts were scarce and levels of self-superparasitism high, suggesting the existence of beneficial effects of self-superparasitism.     

Context-dependent symbiosis between black flies (Diptera: Simuliidae) and trichomycete fungi (Harpellales: Legeriomycetaceae)

The context-dependent nature of a symbiotic relationship between a trichomycete fungus ( Smittium culisetae ) and a larval black fly ( Simulium vittatum ) is demonstrated in the present study. No significant difference was found between the size of larvae colonized by trichomycetes and those free of trichomycetes, regardless of the trichospore dosage or initial age of the larvae. This trend suggests that the trichomycete has no detectable effect on host fitness, indicating a commensalistic relationship. However, in half of the experiments, stressed (i.e. starved) larvae exposed to trichospores at a dosage of 20 000 spores ml⁻¹ had significantly higher survival than did trichomycete-free larvae, indicating a mutualistic relationship. Trichomycetes in adult female black flies can replace the ovaries. The symbiotic association between trichomycetes and simuliids, therefore, is dynamic: commensalistic when larvae are well fed, mutualistic when larvae are starved, and parasitic in adults. The trichomycete-black fly relationship represents a rare case of symbiosis shifting among three states.     

The microdistribution of the trichomycete Smittium culisetae in the hindgut of the black fly host Simulium vittatum

We examined the distribution of hyphae of the trichomycete fungus Smittium culisetae (Harpellales: Legeriomycetaceae) in the hindgut of a larval black fly (Simulium vittatum, cytospecies IS-7) by analyzing its prevalence and relative abundance. Hyphal prevalence was highest in the posterior colon (93.1%) and rectum (86.3%), with low prevalence (12.0%) in the anterior colon. Relative abundance of hyphae was highest in the posterior colon, followed by the rectum; relative abundance of hyphae in the anterior colon was lower. Hyphae of S. culisetae were not observed in the pylorus. We used a novel method of quantifying the relative abundance of S. culisetae in the host hindgut. The hindgut was observed with an ocular grid, and abundance was expressed as the ratio of grids occupied by hyphae to the number of grids occupied by hindgut.     

Trichomycetes living in the guts of aquatic insects of Misiones and Tierra del Fuego, Argentina

Fourteen species of Trichomycetes living in the guts of aquatic insects are reported from two provinces of Argentina, Misiones and Tierra del Fuego. Twelve of the species belong to the Harpellales and two are Amoebidiales. Five harpellid species are reported from Misiones in the extreme northeast of the country (Genistellospora homothallica, Harpella tica, Smittium culisetae, Smittium sp., Stachylina sp.) and seven are from Tierra del Fuego, the southern tip of South America (H. meridianalis, Glotzia sp., S. culicis, S. cellaspora, S. imitatum, Stachylina minima, Penella simulii). Insect hosts all were immature stages of Culicidae, Simuliidae, Chironomidae, Ceratopogonidae (Insecta: Diptera), and Ephemeroptera and Plecoptera. The lower diversity of Trichomycetes found at Misiones, which has a subtropical climate and rainforest vegetation, was due possibly to the warmer temperatures of the water (15-24 C), compared to the colder streams of Tierra del Fuego (9-15 C), with forests and steppes as typical vegetation.     

Behavioral responses of the parasitoid Melittobia digitata to volatiles emitted by its natural and laboratory hosts

Responses of macropterous females of the ectoparasitoid Melittobia digitata Dahms (Hymenoptera: Eulophidae) to direct and indirect cues emitted by its natural hosts as well as laboratory hosts were investigated using a Y‐tube olfactometer. To locate the nest of mud dauber wasps, Trypoxylon politum Say (Hymenoptera: Crabronidae), and one of their inquilines, Anthrax spec., parasitoids exploit volatiles from the freshly built nest mud and the empty cocoon constructed by the wasps, as well as their meconium. However, the parasitoids did not respond to odors emitted by older nest mud or by the host stages that are attacked (T. politum prepupae and Anthrax spec. larvae). Melittobia digitata was not attracted to direct volatiles released by the dipteran hosts Anastrepha ludens Loew (Diptera: Tephritidae) (a natural host) and Sarcophaga bullata (Parker) (Diptera: Sarcophagidae) (a laboratory host). Based on our results, we suggest that M. digitata adopts a ‘sit and wait’ strategy to locate mud dauber wasps, relying mainly on indirect host‐related cues: females search for nests that are under construction and once found, they wait inside the cell until the host completes its cocoon and releases meconium, an indicator that is associated with host suitability. No attraction was found to dipteran hosts, suggesting that parasitization of these hosts may be incidental, due to the broad host plasticity of Melittobia wasps.     

Differential modulation of murine host immune response by salivary gland extracts from the mosquitoes Aedes aegypti and Culex quinquefasciatus

Mosquitoes (Diptera: Culicidae) are major vectors of numerous infectious agents. Compounds in mosquito saliva not only facilitate blood-feeding, but may also have an impact upon the immune system of vertebrate hosts. Consequently, the exposure to mosquito saliva may influence pathogen transmission, establishment and disease development. Using two medically important vector mosquitoes, Aedes aegypti (L.) and Culex quinquefasciatus Say, we examined the effects of mosquito saliva on immune cells of host mice. After antigen-specific or non-specific stimulation, murine splenocyte proliferation and production of both Th1 and Th2 cytokines were significantly reduced in the presence of salivary gland extract (SGE) from Ae. aegypti, but not SGE from Cx. quinquefasciatus. T cell populations were highly susceptible to this suppression, showing increased mortality and reduced division rates - judged by flow cytometric analyses. Evidently these two culicine mosquitoes differ in their host immunomodulatory activities.     

Trifoliellum bioblitzii, a new genus of trichomycete from mayfly nymphs in Nova Scotia, Canada

Trichomycetes are an ecological group of fungi and protists that colonize the gut lining of invertebrates in aquatic and moist terrestrial habitats. The diversity of this group appears to be high with many new species discovered each year. A new genus of fungal trichomycete, Trifoliellum (Harpellales), is described here with the type species T. bioblitzii. This genus is characterized by having unique, trefoil-shaped asexual spores (trichospores). Another new species, Legeriosimilis halifaxensis, also is described from the same mayfly host, Eurylophella temporalis, collected from the same site near Halifax, Nova Scotia.     

Histories of host shifts and cospeciation among free‐living parasitoids of Rhagoletis flies

Host shifts by specialist insects can lead to reproductive isolation between insect populations that use different hosts, promoting diversification. When both a phytophagous insect and its ancestrally associated parasitoid shift to the same novel host plant, they may cospeciate. However, because adult parasitoids are free living, they can also colonize novel host insects and diversify independent of their ancestral host insect. Although shifts of parasitoids to new insect hosts have been documented in ecological time, the long‐term importance of such shifts to parasitoid diversity has not been evaluated. We used a genus of flies with a history of speciation via host shifting (Rhagoletis [Diptera: Tephritidae]) and three associated hymenopteran parasitoid genera (Diachasma, Coptera and Utetes) to examine cophylogenetic relationships between parasitoids and their host insects. We inferred phylogenies of Rhagoletis, Diachasma, Coptera and Utetes and used distance‐based cophylogenetic methods (ParaFit and PACo) to assess congruence between fly and parasitoid trees. We used an event‐based method with a free‐living parasitoid cost model to reconstruct cophylogenetic histories of each parasitoid genus and Rhagoletis. We found that the current species diversity and host–parasitoid associations between the Rhagoletis flies and parasitoids are the primary result of ancient cospeciation events. Parasitoid shifts to ancestrally unrelated hosts primarily occur near the branch tips, suggesting that host shifts contribute to recent parasitoid species diversity but that these lineages may not persist over longer time periods. Our analyses also stress the importance of biologically informed cost models when investigating the coevolutionary histories of hosts and free‐living parasitoids.     

Host range testing of a prospective classical biological control agent against cabbage maggot,Delia radicum,in Canada

The introduction of a European natural enemy, Aleochara bipustulata L. (Coleoptera: Staphylinidae), is being considered for control of cabbage maggot, Delia radicum (L.) (Diptera: Anthomyiidae) in canola in Canada, and the host specificity of this pupal parasitoid must first be studied. Contemporary guidelines were used to select 18 species of Diptera to represent non-target species taxonomically related or ecologically similar to reported hosts of A. bipustulata, or beneficial species. No-choice tests were used to determine which of the 18 species are within A. bipustulata’s fundamental host range, and whether puparial structure or mass or duration of pupal development influence their acceptability and suitability as hosts. Five species were consistently suitable as hosts, and these were either relatively small or were taxonomically closely related to the target host. The probability that a puparium would be entered by a parasitoid larva was greatest for small puparia, but was unaffected by pupal duration. The probability of completing parasitoid development once a puparium was entered was influenced by both puparial mass and duration of pupal development. Pitfall traps to assess habitat associations caught A. bipustulata adults in a variety of crop habitats but none were caught in forests. Host range and habitat data are used to argue that there is little risk of parasitism to beneficial taxa. Non-target species taxonomically closely related to the target D. radicum or with small puparia may fall within the fundamental host range of A. bipustulata. However, risk to many of these species may be minimal because of the habitat preferences of the parasitoid and its cues for host-finding and recognition.     

Nutritional ecology of the interaction between larvae of the gregarious ectoparasitoid, Muscidifurax raptorellus (Hymenoptera: Pteromalidae), and their pupal host, Musca domestica (Diptera: Muscidae)

In this study we examined the relationship between clutch size and parasitoid development of Muscidifurax raptorellus (Hymenoptera: Pteromalidae), a gregarious idiobiont attacking pupae of the housefly, Musca domestica (Diptera: Muscidae). Host quality was controlled in the experiments by presenting female parasitoids with hosts of similar size and age. This is the first study to monitor the development of a gregarious idiobiont parasitoid throughout the course of parasitism. Most female wasps laid clutches of one to four eggs per host, although some hosts contained eight or more parasitoid larvae. In both sexes, parasitoids completed development more rapidly, but emerging adult wasp size decreased as parasitoid load increased. Furthermore, the size variability of eclosing parasitoid siblings of the same sex increased with clutch size. Irrespective of clutch size, parasitoids began feeding and growing rapidly soon after eclosion from the egg and this continued until pupation. However, parasitoids in hosts containing five or more parasitoid larvae pupated one day earlier than hosts containing one to four larvae. The results are discussed in relation to adaptive patterns of host utilization by gregarious idiobiont and koinobiont parasitoids.     

Tripius gyraloura n. sp. (Aphelenchoidea: Sphaerulariidae) parasitic in the gall midge Lasioptera donacis Coutin (Diptera: Cecidomyiidae)

A new nematode, Tripius gyraloura n. sp., is described from the arundo gall midge, Lasioptera donacis Coutin (Diptera: Cecidomyiidae). This gall midge is being considered as a biological control agent for use in North America against the introduced giant reed Arundo donax (L.) (Poaceae: Cyperales). Thus the present study was initiated to investigate a nematode parasite that was unknown at the time studies with L. donacis were initiated. The new species has a rapid development in the fly host and the mature parasitic female nematodes evert their uterine cells in the hosts’ hemolymph. Because large numbers of nematodes sterilise the host, eradication of the parasite from laboratory colonies of the midge may be necessary before populations of the fly are released.     

Fecundity and development of the ectoparasitic waspNasonia vitripennis are dependent on host quality

The gregarious, ectoparasitoidNasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) was offered pupae representing seven fly species, but only members of two families (Sarcophagidae and Muscidae) were parasitized. Host acceptance as an oviposition site did not imply host suitability for parasitoid growth:N. vitripennis produced fewer progeny, a higher proportion of males, required a longer development time, and produced smaller adult wasps onMusca domestica L. (Diptera: Muscidae) than on the three sarcophagid species tested [Sarcophaga bullata Parker,S. crassipalpis Macquart, andPeckia abnormis (Enderlein) (Diptera: Sarcophagidae)]. The physiological and nutritional status of a preferred host,S. bullata, influenced oviposition behavior and development ofN. vitripennis. Progeny allocation and sex ratio, which were regulated by the female parasitoid during oviposition, differed on living and dead nondiapausing hosts and on diapausing pupae. Differences in the host's nutritional condition was reflected in changes of the wasp's development time and adult body size. Envenomation was essential for successful development of the parasitoid on nondiapausing hosts, but venom injection byN. vitripennis did not increase the suitability of diapausing or dead pupae. The results suggest that wasp development is enhanced by changes induced in the host by parasitism.     

Infection success in novel hosts: an experimental and phylogenetic study of Drosophila-parasitic nematodes

Surprisingly little is known about what determines a parasite's host range, which is essential in enabling us to predict the fate of novel infections. In this study, we evaluate the importance of both host and parasite phylogeny in determining the ability of parasites to infect novel host species. Using experimental lab assays, we infected 24 taxonomically diverse species of Drosophila flies (Diptera: Drosophilidae) with five different nematode species (Tylenchida: Allantonematidae: Howardula, Parasitylenchus), and measured parasite infection success, growth, and effects on female host fecundity (i.e., virulence). These nematodes are obligate parasites of mushroom-feeding Drosophila, particularly quinaria and testacca group species, often with severe fitness consequences on their hosts. We show that the potential host ranges of the nematodes are much larger than their actual ranges, even for parasites with only one known host species in nature. Novel hosts that are distantly related from the native host are much less likely to be infected, but among more closely related hosts, there is much variation in susceptibility. Potential host ranges differ greatly between the related parasite species. All nematode species that successfully infected novel hosts produced infective juveniles in these hosts. Most novel infections did not result in significant reductions in the fecundity of female hosts, with one exception: the host specialist Parasitylenchus nearcticus sterilized all quinaria group hosts, only one of which is a host in nature. The large potential host ranges of these parasites, in combination with the high potential for host colonization due to shared mushroom breeding sites, explain the widespread host switching observed in comparisons of nematode and Drosophila phylogenies.     

Effects of linear furanocoumarins on the herbivore Spodoptera exigua and the parasitoid Archytas marmoratus: host quality and parasitoid success

We assessed the effects of three photoactivated linear furanocoumarins, secondary plant metabolites present in Apium spp. (Apiaceae) and other taxa, on the larval-pupal parasitoid, Archytas marmoratus (Townsend) (Diptera: Tachinidae) and the host Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), in the presence and absence of ultraviolet radiation. These results then were compared with previously described responses of Archytas marmoratus and other hosts to different plant allelochemicals. Ultraviolet radiation had no consistent detrimental effects. Increasing concentrations of linear furanocoumarins (from 0 to 0.029% of fresh weight) increased mortality of both the host and parasitoid. Increasing linear furanocoumarin levels also prolonged larval development of surviving S. exigua, but had no effect on pupal developmental time or pupal mass. Consequently, there was no effect of increasing linear furanocoumarin levels on surviving parasitoid development time (from the time of host pupation) or size. These results indicate the effects of linear furanocoumarins are mediated through effects on the host. Because reported responses of Ar. marmoratus to other allelochemicals (e.g., flavones, methyl ketones) are mediated through host effects, these results support the hypothesis that Ar. marmoratus displays a generalized response to plant allelochemicals that are predictable based on host mediated effects.     

Ability of a Generalist Seed Beetle to Colonize an Exotic Host: Effects of Host Plant Origin and Oviposition Host

The colonization of an exotic species by native herbivores is more likely to occur if that herbivore is a generalist. There is little information on the life-history mechanisms used by native generalist insects to colonize exotic hosts and how these mechanisms are affected by host properties. We examined the ability of the generalist seed beetle Stator limbatus Horn to colonize an exotic species. We compared its host preference, acceptability, performance, and egg size when ovipositing and developing on two native (Pithecellobium dulce (Roxb.) Benth and Senegalia riparia (Kunth)) and one exotic legume species (Leucaena leucocephala (Lam.)). We also analyzed the seed chemistry. We found that females recognize the exotic species as an unfavorable host for larval development and that they delayed oviposition and laid fewer and larger eggs on the exotic species than on the native species. Survivorship on the exotic host was 0%. Additionally, seeds of the native species contain five chemical compounds that are absent in the exotic species, and the exotic species contains three sterols, which are absent in the native legumes. Genetically based differences between beetles adapted to different hosts, plastic responses toward new hosts, and chemical differences among seeds are important in host colonization and recognition of the exotic host. In conclusion, the generalist nature of S. limbatus does not influence its ability to colonize L. leucocephala. Explanations for the colonization of exotic hosts by generalist native species and for the success of invasive species must be complemented with studies measuring local adaptation and plasticity.     

Costs of reproduction and geographic variation in the reproductive tactics of the mosquito Aedes triseriatus

Intraspecific host discrimination is widespread in solitary parasitoids whose adult females forage for and evaluate host suitability, whereas interspecific discrimination is less common. In some parasitoid species, mostly Diptera and Coleoptera, the larva performs the last step of host searching. It has been suggested that host discrimination will rarely occur in such host-seeking larvae because their low mobility results in a low host encounter rate. We determined the extent to which the larvae of Aleochara bilineata Gyllenhal (Coleoptera: Staphylinidae), a solitary parasitoid of aggregated Diptera pupae: (1) discriminated between unparasitized hosts and hosts parasitized by conspecifics; (2) used semiochemical cues to discriminate; (3) were influenced by life expectancy, presence of conspecifics and host availability in their host acceptance decision; and the extent to which (4) A. bilineata and A. bipustulata L., a species exploiting the same hosts and occurring sympatrically, showed interspecific host discrimination. A. bilineata larvae were able to discriminate between unparasitized hosts and hosts parasitized by conspecifics in a choice experiment. Such behavior has never previously been described for a coleopteran parasitoid or for a parasitoid species whose larvae perform host searching. Host discrimination in this species was not based on the presence of visual or tactile cues (e.g., entrance holes) but rather on chemical cues. The life expectancy of A. bilineata larvae was significantly shorter in the presence than in absence of hosts, and older larvae had lower parasitism success than young larvae in a 24-h experiment. However, the host acceptance decision of A. bilineata larvae was not influenced by larval age or the presence of conspecifics when the ratio of hosts per larva was greater than or equal to 1. When hosts were scarce, the degree of superparasitism increased significantly with the number of foraging conspecifics and the age of the larvae. Both species of Aleochara showed intra- and interspecific host discrimination in a choice experiment. In contrast to A. bipustulata, A. bilineata larvae more frequently parasitized hosts parasitized by A. bipustulata than those parasitized by conspecifics. We suggest that host discrimination will be frequent in solitary parasitoids with host-seeking larvae when hosts are aggregated. Received: 4 June 1998 / Accepted: 1 September 1998     

Is the host or the parasitoid in control?: effects of host age and temperature on pseudoparasitization by Microplitis rufiventris in Spodoptera littoralis

We compared the production of pseudoparasitization by Microplitis rufiventris females in most (third) and less (fourth) preferred instars of Spodoptera littoralis larvae at 20+/-1 and 27+/-1 degrees C. The parasitized hosts were classified into hosts producing parasitoids (type A hosts) and hosts producing no parasitoids, i.e., pseudoparasitized hosts (type B hosts). The latter were further classified into: (a) pseudoparasitized hosts with "well" arrested development (type B1 hosts); (b) pseudoparasitized hosts with partially arrested development (type B2 hosts); and (c) pseudoparasitized hosts that successfully pupated to apparently normal host pupae (type B3 hosts). The present series of experiments showed that parasitization by M. rufiventris was clearly affected by host instar, age within an instar and rearing temperature. Production of type B hosts was less when third instar S. littoralis larvae were exposed to the wasp females than when the host larvae were in fourth instar. The production of type A hosts was much greater when early or mid ages of an instar was stung by the wasp females comparing with stung late age of the same instar. Production of type B hosts may be due to one or overall of the following: (a) dosage dilution of M. rufiventris female's factors in the different age classes of the instar; (b) endocrine system (physiological state) at parasitization time, i.e., early vs late age of the instar; (c) growth rate of host larvae. The lowest production of type B hosts was at highest growth rate; and (d) temperature, larger proportions of type B hosts were produced at 27+/-1 than at 20+/-1degrees C. The three types host development (B1, B2 and B3) are possibly representing three levels of host resistance (host control) resulting in partial or complete failure of parasitoid control. Type A hosts represent complete success of parasitoid control. The results suggest that the impact of parasitoid factor(s) on developmental arrest is affected by host age at the time of parasitism and/or by temperature.