Biological control of gypsy moth (Lymantria dispar) by the entomopathogenic fungus Entomophaga maimaiga in Bulgaria in 2021

In 2021, biological control programme against gipsy moth (Lymantria dispar) populations was carried out by introduction of the entomopathogenic fungus Entomophaga maimaiga on the territory of four State Forest Enterprises: Municipal Enterprise (ME) ‘Management of Municipal Forests, Agriculture and Forestry’, Nessebar; State Game Enterprises (SGE) Nessebar and Balchik; State Forestry (SF) Vidin. The pathogen was introduced during the period 15-26.03.2021 in 34 localities - five in ME Nessebar, eight in SGE Nessebar, ten in SGE Balchik and eleven in SF Vidin. The average number of gypsy moth population density in the locations of introduction was relatively high, ranging between 0.4-15.9 egg mass/tree in the area of ​​SGE Balchik and 11.9-65.0 egg mass/tree in the area of ​​ME Nessebar. The average mortality of young gypsy moth caterpillars (first-third instar) due to E. maimaiga varied between 2.6% (SGE Balchik) and 13.0% (SF Vidin), and of caterpillars in later fourth-sixth instar - between 20.7% (SF Vidin) and 52.4% (ME Nessebar). The overall mortality of the gipsy moth caterpillars due to E. maimaiga was lowest in the region of SGE Balchik (26.1%), followed by SF Vidin (33.7%), SGE Nessebar (48.5%) and ME Nessebar (55.9%). As a result of the introduction, gipsy moth severe outbreaks in the region of Nessebar was significantly suppressed. The high number of E. maimaiga resting spores persists in the surface layers of the soil in the other two areas (Vidin and Balchik) has the potential to suppress L. dispar attacks in next years.

Successful Completion of an International Project of the Department of Natural Sciences at New Bulgarian University

The paper presents the results of a successfully completed international project of the Department of Natural Sciences titled “Monitoring and identification of the entomopathogenic fungus Entomophaga maimaiga in Lymantria dispar populations” and funded by the National Research Fund in 2017-2019. Investigating the distribution of E. maimaiga in Austria and Bulgaria in 2018-2019 shows that the pathogen has expanded its range to the west and reached Lower Austria. E. maimaiga has been identified in Bulgaria as well and has become part of the natural enemy complex of L. dispar.

Occurrence of gypsy moth (Lymantria dispar L.) in the Slovak Republic and its outbreaks during 1945–2020

The gypsy moth is one of the most serious pests in forests and fruit tree plantations over prevailing parts of the Northern Hemisphere. This work is based on a literature review, and presents history of gypsy moth Lymantria dispar L., observed in Slovak forests within the period 1945–2020. The life cycle, hosts, natural enemies, population dynamics of pests, impact of outbreaks on forests and different management methods used in the past are discussed. Since 1945, there were nine gypsy moth outbreaks in Slovakia. Between 1945 and 2020, a total of 155,034 ha of deciduous forests were touched with varying intensity, representing an average annual damage of 2,040 ha. The strongest outbreak culminated in 2004. Totally 51,479 ha were attacked in the period of 2000–2008. We have found outbreak periods that repeat with frequency of 7.8 ±2.2 years and the average outbreak phase lasts 3.1 ±1.1 years. The period between two subsequent outbreaks seems to be more or less constant and duration of the outbreak phase seems to be gradually shortened during the study period. Several factors influencing the gypsy moth population dynamics in Slovakia are discussed. The role of biological control by using entomopathogenic fungus Entomophaga maimaiga is described.

Relating Aerial Deposition of Entomophaga maimaiga Conidia (Zoopagomycota: Entomophthorales) to Mortality of Gypsy Moth (Lepidoptera: Erebidae) Larvae and Nearby Defoliation

We collected data on mortality of late-instar gypsy moth, Lymantria dispar (L.), from outbreak populations over 4 wk in June 2017 at 10 sites in the New England region of the United States, along with estimated rainfall at these sites. Deposition of airborne conidia of the fungal pathogen, Entomophaga maimaiga Humber, Shimazu & R.S. Soper, was measured at these same sites as well as at seven other locations in New England. We also quantified the geographical distribution of gypsy moth-caused defoliation in New England in 2017 and 2018 from Landsat imagery. Weekly mortality of gypsy moth larvae caused by E. maimaiga correlated with local deposition of conidia from the previous week, but not with rainfall. Mortality from this pathogen reached a peak during the last 2 wk of gypsy moth larval development and always exceeded that caused by LdNPV, the viral pathogen of gypsy moth that has long been associated with gypsy moth outbreaks, especially prior to 1989. Cotesia melanoscela (Ratzeburg) was by far the most abundant parasitoid recovered and caused an average of 12.6% cumulative parasitism, but varied widely among sites. Deposition of E. maimaiga conidia was highly correlated with percent land area defoliated by gypsy moths within distances of 1 and 2 km but was not significantly correlated with defoliation at distances greater than 2 km. This is the first study to relate deposition of airborne conidia of E. maimaiga to mortality of gypsy moths from that agent.

Testing of soil distribution, viability, and infectivity of azygospores of the entomopathogenic fungus Entomophaga maimaiga

Entomophaga maimaiga is one of the best-known gypsy moth pathogens with increasing importance in the biological control aimed at reducing the population of this defoliator. The territory of the Republic of Serbia is a very favorable gypsy moth habitat, and the problems caused by its larvae during the periods of outbreaks are so great that they can lead to forest defoliation and forest health deterioration. In the Republic of Serbia, E. maimaiga was first recorded in 2011 (as a new species in the microflora of Serbia) during the gypsy moth outbreak (2009-2014), which was the last to date. In 2011, numerous dead caterpillars were found on and around trees in oak and beech stands of the Belgrade and Valjevo regions in Serbia. Subsequent laboratory analyses revealed that they contained azygospores and conidia of this fungus. The assisted introduction of E. maimaiga was carried out in Compartments 28 and 30 of the FMU Avala on Mt. Avala in 2011. It led to an epizootic in the following year and stopped the outbreak of the gypsy moth in the area. Since it has been seven years since the outbreak, we tested the viability and infectivity of E. maimaiga azygospores obtained from the soil samples from Avala Mountain where its assisted introduction was carried out. We used larvae from the laboratory gypsy moth population and applied the bioassay method. The results show that azygospores are present in the given soil samples, they are viable and their infectivity is conserved. It can be concluded that in the case of an outbreak of gypsy moth populations in the future, they will exhibit their entomopathogenic effect

Sleeping Beauties: Horizontal Transmission via Resting Spores of Species in the Entomophthoromycotina

Many of the almost 300 species of arthropod-pathogenic fungi in the Entomophthoromycotina (Zoopagomycota) are known for being quite host-specific and are able to cause epizootics. Most species produce two main types of spores, conidia and resting spores. Here, we present a review of the epizootiology of species of Entomophthoromycotina, focusing on their resting spores, and how this stage leads to horizontal transmission and persistence. Cadavers in which resting spores are produced can often be found in different locations than cadavers of the same host producing conidia. Resting spores generally are dormant directly after production and require specific conditions for germination. Fungal reproduction resulting from infections initiated by Entomophaga maimaiga resting spores can differ from reproduction resulting from conidial infections, although we do not know how commonly this occurs. Reservoirs of resting spores can germinate for variable lengths of time, including up to several months, providing primary infections to initiate secondary cycling based on conidial infections, and not all resting spores germinate every year. Molecular methods have been developed to improve environmental quantification of resting spores, which can exist at high titers after epizootics. Ecological studies of biological communities have demonstrated that this source of these spores providing primary inoculum in the environment can decrease not only because of germination, but also because of the activity of mycopathogens.

Modification of a Pollen Trap Design To Capture Airborne Conidia of Entomophaga maimaiga and Detection of Conidia by Quantitative PCR

ABSTRACT The goal of this study was to develop effective and practical field sampling methods for quantification of aerial deposition of airborne conidia of Entomophaga maimaiga over space and time. This important fungal pathogen is a major cause of larval death in invasive gypsy moth (Lymantria dispar) populations in the United States. Airborne conidia of this pathogen are relatively large (similar in size to pollen), with unusual characteristics, and require specialized methods for collection and quantification. Initially, dry sampling (settling of spores from the air onto a dry surface) was used to confirm the detectability of E. maimaiga at field sites with L. dispar deaths caused by E. maimaiga, using quantitative PCR (qPCR) methods. We then measured the signal degradation of conidial DNA on dry surfaces under field conditions, ultimately rejecting dry sampling as a reliable method due to rapid DNA degradation. We modified a chamber-style trap commonly used in palynology to capture settling spores in buffer. We tested this wet-trapping method in a large-scale (137-km) spore-trapping survey across gypsy moth outbreak regions in Pennsylvania undergoing epizootics, in the summer of 2016. Using 4-day collection periods during the period of late instar and pupal development, we detected variable amounts of target DNA settling from the air. The amounts declined over the season and with distance from the nearest defoliated area, indicating airborne spore dispersal from outbreak areas. IMPORTANCE We report on a method for trapping and quantifying airborne spores of Entomophaga maimaiga, an important fungal pathogen affecting gypsy moth (Lymantria dispar) populations. This method can be used to track dispersal of E. maimaiga from epizootic areas and ultimately to provide critical understanding of the spatial dynamics of gypsy moth-pathogen interactions.