Effects of the ant Formica fusca on the transmission of microsporidia infecting gypsy moth larvae

Transmission plays an integral part in the intimate relationship between a host insect and its pathogen that can be altered by abiotic or biotic factors. The latter include other pathogens, parasitoids, or predators. Ants are important species in food webs that act on various levels in a community structure. Their social behavior allows them to prey on and transport larger prey, or they can dismember the prey where it was found. Thereby they can also influence the horizontal transmission of a pathogen in its host's population. We tested the hypothesis that an ant species like Formica fusca L. (Hymenoptera: Formicidae) can affect the horizontal transmission of two microsporidian pathogens, Nosema lymantriae Weiser (Microsporidia: Nosematidae) and Vairimorpha disparis (Timofejeva) (Microsporidia: Burenellidae), infecting the gypsy moth, Lymantria dispar L. (Lepidoptera: Erebidae: Lymantriinae). Observational studies showed that uninfected and infected L. dispar larvae are potential prey items for F. fusca. Laboratory choice experiments led to the conclusion that F. fusca did not prefer L. dispar larvae infected with N. lymantriae and avoided L. dispar larvae infected with V. disparis over uninfected larvae when given the choice. Experiments carried out on small potted oak, Quercus petraea (Mattuschka) Liebl. (Fagaceae), saplings showed that predation of F. fusca on infected larvae did not significantly change the transmission of either microsporidian species to L. dispar test larvae. Microscopic examination indicated that F. fusca workers never became infected with N. lymantriae or V. disparis after feeding on infected prey.     

Modeling horizontal transmission of microsporidia infecting gypsy moth,Lymantria dispar (L.), larvae

We developed a simulation model that describes the horizontal transmission of three different microsporidia, Endoreticulatus schubergi, Nosema lymantriae and Vairimorpha disparis and their insect host, the gypsy moth, Lymantria dispar. The model describes the stage specific development and mortality of uninfected, latently infected or infectious hosts, the food consumption, the infection by spore-laden feces of E. schubergi and N. lymantriae and by spore-laden cadaver of N. lymantriae and V. disparis. Model results were compared to percent infection of L. dispar test larvae published in earlier studies using caged oak trees and potted oak-plants. When feces were selected as the source of spores for transmission of E. schubergi or N. lymantriae, the model estimated a percent infection in susceptible larvae that was in the range of the experimental studies. When spore-laden cadavers were the source of spores of N. lymantriae or V. disparis, the model did not correctly predict the experimentally measured percent infection in susceptible larvae. The most critical points of the simulation model are exact calculation of spore release, mortality and exact determination of the transmission coefficients when cadavers were included as a source for microsporidian infection.     

Three microsporidian pathogens infecting Lymantria dispar larvae do not differ in their success in horizontal transmission

We quantified horizontal transmission of three microsporidian pathogens, Endoreticulatus schubergi, Nosema lymantriae and Vairimorpha disparis that infect Lymantria dispar larvae in an experiment using caged, potted oak plants. Despite marked differences in the modes of spore release from infectious hosts, no significant differences in the transmission success to uninfected, susceptible test hosts were ascertained between the tested microsporidian species. The density of initially inoculated larvae and the exposure period, on the other hand, did influence the number of infected test larvae. Depending on the density of inoculated larvae (10%, 30% or 50%), between 0% and 26% of the test larvae became infected with one of the three tested microsporidian pathogens after an exposure period of 6 days. When the exposure period was 12 days, between 11% and 76% of the test larvae became infected.     

Vertical transmission and overwintering of microsporidia in the gypsy moth, Lymantria dispar

Vertical transmission and the overwintering success of three different microsporidia infecting Lymantria dispar (Lepidoptera: Lymantriidae) larvae were investigated. Endoreticulatus schubergi, a midgut pathogen, was transmitted to offspring via female and male via the egg chorion (transovum transmission). Between 8% and 29% of the emerging larvae became infected. No spores of E. schubergi were found in surface-washed eggs. Nosema lymantriae, a microsporidium that causes systemic infections, was transovarially transmitted. Between 35% and 72% of the progeny were infected. Vairimorpha disparis, a fat body pathogen, was not vertically transmitted. The infectivity of spores that overwintered in cadavers of infected L. dispar varied by species, placement in the environment, and weather conditions. Spores of E. schubergi were still infective after an eight month exposure period of cadavers on the ground. Spores of N. lymantriae and V. disparis remained highly infective only when cadavers overwintered under a more or less continuous snow cover for four months.     

Host specificity of microsporidia pathogenic to the gypsy moth, Lymantria dispar (L.): Field studies in Slovakia

Several species of microsporidia are important chronic pathogens of Lymantria dispar in Europe but have never been recovered from North American gypsy moth populations. The major issue for their introduction into North American L. dispar populations is concern about their safety to native non-target insects. In this study, we evaluated the susceptibility of sympatric non-target Lepidoptera to two species of microsporidia, Nosema lymantriae and Vairimorpha disparis, isolated from European populations of L. dispar and applied in field plots in Slovakia. Application of ultra low volume sprays of the microsporidia maximized coverage of infective spores in a complex natural environment and, thus, exposure of non-target species to the pathogens. Of 653 non-target larvae collected from plots treated with V. disparis in 2002, 18 individual larvae representing nine species in four families were infected. These plots were monitored for two subsequent seasons and V. disparis was not recovered from non-target species. Of 2571 non-target larvae collected in N. lymantriae-treated sites, one larva was found to be infected. Both species of microsporidia, particularly N. lymantriae, appear to have a very narrow host range in the field, even when an inundative technique is used for their introduction. V. disparis infections in L. dispar exceeded 40% of recovered larvae in the treated study sites; infection rates were lower in sites sprayed with N. lymantriae. Several naturally-occurring pathogens were recorded from the non-target species. The most common pathogen, isolated from 21 species in eight families, was a microsporidium in the genus Cystosporogenes.     

Adult feeding affects fecundity of the predator,Calosoma sycophanta (Col.: Carabidae)

Calosoma sycophanta L. adults were fed either gypsy moth (Lymantria dispar L.) larvae or split grapes for set periods of time while their reproduction was monitored. Few female beetles reproduced unless fed gypsy moth larvae during the first week after they ended hibernation. Even females initially fed grapes that were later fed larvae had reduced reproduction. The implications these results have for relationships between beetle and gypsy moth populations are discussed.     

Influence of Dimilin on a microsporidian infection in the gypsy moth Lymantria dispar (L.) (Lepidoptera: Lymantriidae)

Bioassay studies were conducted to investigate the influence of Dimilin (diflubenzuron), a chitinsynthetase inhibitor used for insecticidal control of the gypsy moth, Lymantria dispar, on the development and viability of a microsporidian pathogen of L. dispar. Before or after an infection with a Nosema species, L. dispar larvae were fed Dimilin in sublethal dosages. Dimilin fed to L. dispar larvae at 0.65 ng/cm² diet surface resulted in a total larval mortality of 53%. Although the microsporidian infection alone did not cause high mortality rates (9%), mortality increased to 96% when L. dispar larvae were inoculated with both Dimilin and Nosema spores. When Dimilin was fed to the larvae 24 h before or 6 days after inoculation with the microsporidium, the number of mature spores produced was significantly reduced. When Dimilin was fed to the larvae 24 h after microsporidian inoculation, the number of spores produced was not significantly reduced. Spores that were produced in larvae after Dimilin had been ingested with the diet were less infectious than spores produced in control larvae; the experimental infection rate decreased from 94% when spores obtained from control larvae were used, to 48 or 10% when spores obtained from larvae fed Dimilin 24 h or 6 days after Nosema inoculation, respectively, were used. Mature microsporidian spores washed in Dimilin solution prior to oral inoculation, however, were as infectious as spores stored in liquid nitrogen. We have shown that Dimilin interferes with the establishment of the parasite in its host. In addition, when Nosema sp. succeeds in infecting the L. dispar host despite treatment with Dimilin, the microsporidium does not develop optimally and spore production is reduced.     

Quantifying horizontal transmission of Nosema lymantriae, a microsporidian pathogen of the gypsy moth, Lymantria dispar (Lep., Lymantriidae) in field cage studies

Nosema lymantriae is a microsporidian pathogen of the gypsy moth, Lymantria dispar that has been documented to be at least partially responsible for the collapse of L. dispar outbreak populations in Europe. To quantify horizontal transmission of this pathogen under field conditions we performed caged-tree experiments that varied (1) the density of the pathogen through the introduction of laboratory-infected larvae, and (2) the total time that susceptible (test) larvae were exposed to these infected larvae. The time frame of the experiments extended from the early phase of colonization of the target tissues by the microsporidium to the onset of pathogen-induced mortality or pupation of test larvae. Upon termination of each experiment, the prevalence of infection in test larvae was evaluated. In the experiments performed over a range of pathogen densities, infection of test larvae increased with increasing density of inoculated larvae, from 14.2 ± 3.5% at density of 10 inoculated per 100 larvae to 36.7 ± 5.7% at 30 inoculated per 100 larvae. At higher densities, percent infection in test larvae appeared to level off (35.7 ± 5.5% at 50 inoculated per 100 larvae). When larval exposure to the pathogen was varied, transmission of N. lymantriae did not occur within the first 15 d post-inoculation (dpi) (11 d post-exposure of test larvae to inoculated larvae). We found the first infected test larvae in samples taken 20 dpi (16 d post-exposure). Transmission increased over time; in the cages sampled 25 dpi (21 d post-exposure), Nosema prevalence in test larvae ranged from 20.6% to 39.2%.     

Herbivory meets fungivory: insect herbivores feed on plant pathogenic fungi for their own benefit

Plants are regularly colonised by fungi and bacteria, but plant‐inhabiting microbes are rarely considered in studies on plant–herbivore interactions. Here we show that young gypsy moth (Lymantria dispar) caterpillars prefer to feed on black poplar (Populus nigra) foliage infected by the rust fungus Melampsora larici‐populina instead of uninfected control foliage, and selectively consume fungal spores. This consumption, also observed in a related lepidopteran species, is stimulated by the sugar alcohol mannitol, found in much higher concentration in fungal tissue and infected leaves than uninfected plant foliage. Gypsy moth larvae developed more rapidly on rust‐infected leaves, which cannot be attributed to mannitol but rather to greater levels of total nitrogen, essential amino acids and B vitamins in fungal tissue and fungus‐infected leaves. Herbivore consumption of fungi and other microbes may be much more widespread than commonly believed with important consequences for the ecology and evolution of plant–herbivore interactions.     

Variation in the ability of larvae of phytophagous insects to develop on evolutionarily unfamiliar plants: a study with gypsy moth Lymantria dispar and Eucalyptus

• 1 By examining variation in the abilities of polyphagous insects to develop on host plants with secondary metabolites that they have never encountered previously, we may be able to gain some insights into the nature of evolution of biochemical mechanisms to process plant secondary metabolites by phytophagous insects.
• 2 The present study aimed to examine variation in the ability of gypsy moth larvae Lymantria dispar (Lymantriidae) to complete development on different species of the plant genus Eucalyptus (Myrtaceae). Leaves of at least some Eucalyptus species contain formylated phloroglucinol derivatives. These are secondary metabolites that are evolutionarily unfamiliar to the gypsy moth.
• 3 Larvae of gypsy moth showed extremely variable responses in larval performance between Eucalyptus species, between individual trees within host plant species, between moth populations, and between individuals within moth populations.
• 4 Larval survivorship was in the range 0–94%, depending on the host. Failure of at least some larvae to complete development on some Eucalyptus species indicates that gypsy moth larvae have a limited ability to process secondary metabolites in eucalypt leaves.
• 5 At least some individuals, however, appear to already possess biochemical mechanisms that process the secondary metabolites in leaves of Eucalyptus species, and therefore the abilities of larvae to complete development on phylogenetically and chemically unfamiliar hosts are already present before the gypsy moth encounters these potential hosts.

     

Modeling the Influence of Rainfall and Temperature on the Phenology of Infection of Gypsy Moth, Lymantria dispar, Larvae by the Fungus Entomophaga maimaiga

A computer model driven by daily maximum-minimum temperature and rainfall records was developed to investigate the influence of weather on times when gypsy moth larvae, Lymantria dispar, can become infected by the fungal pathogen Entomophaga maimaiga. In the model, gypsy moth eggs are hatched and neonates are exposed primarily to germinating resting spores in the soil during the spring. Risk of infection is related to forest floor moisture. Larval and fungal development follows a degree-day model. When larvae become 4th instars, they can again become infected by resting spores because they hide in the litter during daylight hours. If rain falls when infected caterpillars die, the fungus sporulates, producing conidia. The number of conidia produced is assumed to directly influence the probability of infection of other larvae. The model was run using weather records from 1990 through 1992. Predicted times of infection were compatible with estimated fungal recruitment rates and changes in field disease prevalence rates. Assumptions about infection mechanisms in the model are discussed as they relate to the real world.     

First record of Entomophaga maimaiga (Entomophthorales: Entomophthoraceae) in Slovakia

The entomopathogenic fungus Entomophaga maimaiga was found for the first time in Slovakia in 2013. Late instar larvae of gypsy moth, Lymantria dispar, from two sites with different population densities were dissected to evaluate the presence of pathogens. The presence of conidia and resting spores of E. maimaiga in gypsy moth cadavers was confirmed from both sites.     

Estimating the susceptibility of tree species to attack by the gypsy moth, Lymantria dispar

ABSTRACT.

• 1 Numbers of gypsy moth larvae feeding on each of 922 randomly sampled trees in a Quercus—Acer—Fraxinus forest in southwestern Quebec, Canada were counted in 1979 and in 1980 to quantify the larval feeding preferences as observed in the field for eighteen deciduous and one coniferous tree species at the northern range limit of the gypsy moth.
• 2 Both the diameter at breast height (dbh) and the estimated foliage biomass of the sampled trees were used to calculate the relative proportions of foliage represented by each of the nineteen tree species in the forest canopy. With these data on availability and utilization of the tree species by the gypsy moth larvae an Ivlev-type electivity index was used to quantify the larval feeding preferences. These preferences observed in the field define the susceptibility of a tree species to attack by the gypsy moth.
• 3 The feeding preferences calculated using estimated foliage biomass were comparable to the simpler calculation based on dbh (Spearman's rho = 0.79; P= 0.0001). The dbh-based feeding preferences remained almost unchanged in 1979 and 1980 (Spearman's rho = 0.83; P= 0.0001).
• 4 The composite 1979—80, dbh-based feeding preferences show Quercus rubra, Populus grandidentata, Ostrya virginiana, Amelanchier spp. and Acer saccharum were preferentially attacked by gypsy moth. Prunus serotina, Betula lutea, Acer rubrum, A. pensylvanicum, Fraxinus americana, Ulmus rubra, P. pensylvanicum and B. papyrifera were avoided. All nineteen tree species were, however, utilized to at least some degree by gypsy moth larvae.
• 5 These results quantitatively affirm and clarify earlier reports of gypsy moth feeding preferences in North America and Eurasia. The advantages and limitations of using an electivity index to estimate the susceptibility of different tree species to attack by folivores like the gypsy moth are discussed.

     

Two different and sublethal isolates of Nosema lymantriae (Microsporidia) reduce the reproductive success of their host,Lymantria dispar

We examined the effects of two microsporidian isolates of Nosema lymantriae (Germany isolate; Schweinfurt isolate) on the reproductive success of Lymantria dispar L. All possible mating combinations were tested. Both isolates affected the fecundity of infected females and the hatch of neonates. The infection of female L. dispar with either isolate resulted in a higher proportion of non-viable eggs; the survival of neonates during egg stage was not affected. When L. dispar larvae were infected with N. lymantriae [Germany] the number of eggs per egg mass decreased between 24 and 61%. When both adults were infected, the hatch rate decreased to 26%. While the infection of the male or the female host with the Germany isolate affected the number of eggs per egg mass and the hatch of progeny, we did not find a significant effect when male hosts were infected with the Schweinfurt isolate; only infection of the female L. dispar resulted in a reduction of the number of eggs per egg mass between 26 and 37%.     

Food utilization values of gypsy moth Lymantria dispar (Lepidoptera: Lymantriidae) larvae infected with the microsporidium Vairimorpha sp. (Microsporidia: Burenellidae)

Infection of the gypsy moth, Lymantria dispar, with the microsporidium Vairimorpha sp. strongly influences the development of the host in ways typical of many species of terrestrial entomopathogenic Microsporidia; growth is reduced while development time is extended in infected insects. The appearance of the different stages of the parasite in the host relative to the elapsed time after oral infection, as well as the influence of the parasite proliferation on food utilization of the host, were examined. At 3 days postinfection, midgut muscle cells were infected with primary spores, and the fat body tissues contained meronts, sporonts, and primary spores. Many more fat body cells contained vegetative stages and primary spores at 4 and 5 days postinfection, and diplokaryotic spores and immature octospores were also present. Approximate digestibility of infected larvae increased during this time period, whereas the conversion of ingested and digested food to body substance decreased. The relative growth rate of infected and uninfected groups did not differ significantly between 4 and 5 days postinfection, although the relative consumption rate in infected L. dispar larvae was higher. Between 8 and 10 days postinfection, the relative growth rate of uninfected larvae increased. The infected group did not demonstrate this increase at a time period characterized by maturation of diplokaryotic spores and octospores in larval fat body tissues. Total body weight of uninfected larvae remained higher than that of infected larvae after 8 days postinfection.     

Dissemination of the biocontrol agent Vairimorpha necatrix by the spined soldier bug, Podisus maculiventris

The ability of the spined soldier bug, Podisus maculiventris (Say) (Heteroptera: Pentatomidae), to disseminate infective forms of two lepidopteran pathogens, Vairimorpha necatrix (Kramer) (Microspora: Microsporidia) and Lacanobia oleracea granulovirus (LoGV) was investigated. Individual female P. maculiventris that had fed on Lacanobia oleracea L. (Lepidoptera: Noctuidae) larvae, infected with V. necatrix, excreted approximately 6 × 10⁸V. necatrix spores during the subsequent 7 days. Excreted spores were fed to L. oleracea larvae, causing 100% mortality, indicating that the spores remained viable after passing through the gut of the predator. Podisus maculiventris that had fed on V. necatrix or LoGV‐infected larvae were allowed to defecate on the foliage of tomato plants, prior to the infestation of the plants with L. oleracea or Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) larvae. This proved to be an effective way of infecting the pest larvae with the pathogens, particularly when five predatory bugs were used per plant. After 20 days, the number of S. littoralis and L. oleracea surviving on the plants was reduced by 75% and 61%, respectively. Female P. maculiventris maintained on V. necatrix‐infected prey showed reduced egg production and longevity, whilst those fed on LoGV‐infected prey showed only reduced egg production. The potential for P. maculiventris to disseminate insect pathogens is discussed in the context of improved biological control of lepidopteran pests.     

L'applicazione di un virus poliedrico nucleare nella lotta control larve diLymantria dispar L.

Summary Small scale field tests with a nuclear-polyhedrosis virus were conducted, for the first time in Sardinia during 1965–1966, on the larvae of the gypsy moth (Lymantria dispar L.). Promising results were obtained by spraying the pathogen at several dosage levels on the cork-oak trees (Quercus suber L.). Mortality varied with the concentration of virus used and with the age of the larvae at the time of spraying. In 1965 a suspension containing 1,2×10⁹ purified polyhedra per milliliter gave 100% mortality of the second-instar larvae in 16 days. Lower polyhedra counts gave mortality ranging between 84 and 99 per cent. In 1966 the treatments with a raw suspension containing 10⁸ polyhedra per milliliter brought about the death, in 20 days, of 100 per cent and 93 per cent respectively of the second- and third-instar larvae. Even lower concentrations caused considerable larval mortality. The lower temperatures occurring in 1966 account for the delay in the appearance of elevated mortality rates. Application of the virus to third-instar larvae carrying microsporidian infections (Nosema lymantriae Weiser) led to an accelerated mortality. On the basis of the frass collected it is obvious that the insects fed poorly on the treated trees as compared to the controls. It is not possible to make direct comparisons of dosages between the 1965 and the 1966 treatments. The different feeding activity, however, of second-instar larvae treated with similar concentrations of purified or unpurified polyhedra suggest that further research is necessary in order to ascertain if the frass reduction obtained using the raw virus suspension is not due to a greater efficacy of the unpurified as compared to the purified virus. Field results indicate that a virus suspension containing 10 million polyhedral inclusion bodies per milliliter of water will prevent excessive defoliation of trees by the larvae.      

Horizontal and vertical transmission of a Nosema sp. (Microsporidia) from Lymantria dispar (L.) (Lepidoptera: Lymantriidae)

The gypsy moth, Lymantria dispar L. (Lepidoptera, Lymantriidae), a serious defoliator of deciduous trees, is an economically important pest when population densities are high. Outbreaking populations are, however, subject to some moderating influences in the form of entomopathogens, including several species of microsporidia. In this study, we conducted laboratory experiments to investigate the transmission of an unusual Nosema sp. isolated from L. dispar in Schweinfurt, Germany; this isolate infects only the silk glands and, to a lesser extent, Malpighian tubules of the larval host. The latent period ended between 8 and 15 days after oral inoculation and spores were continuously released in the feces of infected larvae until pupation. Exclusion of feces from the rearing cages resulted in a 58% decrease in horizontal transmission. The silk of only 2 of 25 infected larvae contained microsporidian spores. When larvae were exposed to silk that was artificially contaminated with Nosema sp., 5% became infected. No evidence was found for venereal or transovum (including transovarial) transmission of this parasite.     

Effects of elicitation treatment and genotypic variation on induced resistance in Populus: impacts on gypsy moth (Lepidoptera: Lymantriidae) development and feeding behavior

We examined the effects of various wounding treatments and genotypic variation on induced resistance in Populus (Salicales: Salicaceae) against herbivory by the gypsy moth, Lymantria dispar L. (Lepidoptera: Lymantriidae). Second-instar larvae grew and consumed less on leaves from induced than non-induced trees. Likewise, larvae preferred leaf disks from non-induced trees. Among induction treatments, gypsy moth feeding had the strongest and most consistent effect in behavioral choice tests. Mechanical wounding of leaves and mechanical wounding plus application of gypsy moth regurgitant had intermediate effects, while application of jasmonic acid had the weakest overall effect. Under no-choice conditions, there were no consistent trends across clones in the ability of various treatments to elicit plant responses affecting the herbivore. Levels of constitutive and inducible resistance to herbivory varied significantly among 12 Populus clones. Larvae grew up to 30-fold more, and consumed up to 250-fold more on the most suitable than the least suitable clone. Prior feeding by gypsy moths reduced larval feeding up to 71.4% on the most highly inducible clone, but it had little or no effect for the least inducible clones. There was no evidence for a relationship between levels of inducible and constitutive resistance, or between inducible resistance and phylogenetic relatedness among clones. We discuss implications for the ecology and evolution of plant-insect interactions and the management of insect pests. Received: 12 October 1998 / Accepted: 22 March 1999     

Prey‐processing by avian predators enhances virus transmission in the gypsy moth

The Lymantria dispar nucleopolyhedrovirus (LdNPV) is one of the most important regulators of gypsy moth populations, but some aspects of its transmission remain poorly understood, particularly its high rate of spatial spread and ability to persist in low‐density populations. We tested the role of predatory birds in the transmission of this virus using experimental gypsy moth populations in an aviary. Predatory birds captured virus‐infected caterpillars and facilitated viral dispersal via two processes: 1) by ingesting infected caterpillars and passing viral occlusion bodies (OBs) through their guts, and 2) by scattering OBs during predator‐specific processing behaviors, a mechanism documented here for the first time. The relative importance of both pathways differed by predator species. After eating virus‐infected gypsy moth larvae, red‐eyed vireos and black‐capped chickadees passed more gypsy moth nucleopolyhedrovirus in feces than did gray catbirds. During prey‐processing, the repetitive beating of caterpillars by red‐eyed vireos, a behavior that was rarely utilized by chickadees and catbirds, resulted in the scattering of infectious virus. Due to the combination of efficient gut passage and virus spread from prey beating, higher rates of transmission occurred in experimental gypsy moth populations exposed to red‐eyed vireos than those exposed to catbirds or chickadees. Our results show that effective virus transmission was achieved when virus was vectored by predatory birds through a combination of both behavioral and physiological traits.