Microsporidia-cypovirus interactions during simultaneous infection of the tree defoliator Dendrolimus sibiricus (Lepidoptera: Lasiocampidae).

The Siberian moth, Dendrolimus sibiricus is a dangerous forest defoliator, the number one pest of boreal forests in Asia. Search for effective and ecologically friendly control measures drives attention to microbial pathogens. Viruses and microsporidia are obligate intracellular parasites widespread in insect populations causing either chronic or acute infections. Interactions of these pathogens vary from antagonistic to synergistic. The goal of the work was to test a recently discovered cytoplasmatic polyhedrosis virus (cypovirus) strain DsCPV-1 isolated from D.sibiricus, combined with a microsporidium, against D. sibiricus, by feeding the inoculum (viral polyhedral and microsporidian spores). Three different microsporidian parasites of lepidopterans were tested against D. sibiricus as monoinfection: Nosema bombycis from silkworm, N. pyrausta from corn borer, and Tubulinosema loxostegi from beet webworm. Nosema bombycis was the most virulent, with a median lethal time of 7 days in the first and second instars treated with 100,000 and 1 million spores/larva, respectively. Nosema bombycis (dose 100,000 spores/larva) was chosen to test it as mixed infection in combination with an extremely low dose of DsCPV-1 (1 polyhedron/larva) against two races of D. sibiricus second instar larvae (the fir-feeding race and the larch-feeding race). The mixed infection demonstrated the most prominent negative effect on larval lethal time and weight for the both tested races. Mixed infections showed a synergistic effect for the fir-feeding larvae but additive effect only for the larch feeding larvae. Both pathogens co-developed successfully in the larvae with equal ratio of producing inoculum. The combination of these entomopathogens is therefore promising for forest protection against the Siberian moth and could be the way to significantly decrease the amount of pathogens applied in field.

A TaqMan Assay for the Detection and Monitoring of Potentially Invasive Lasiocampids, With Particular Attention to the Siberian Silk Moth, Dendrolimus sibiricus (Lepidoptera: Lasiocampidae).

The Siberian silk moth, Dendrolimus sibiricus Tschetverikov, is a very serious pest of conifers in Russia and is an emerging threat in North America where an accidental introduction could have devastating impacts on native forest resources. Other Dendrolimus Germar species and related Eurasian lasiocampids in the genus Malacosoma (Hubner) could also present a risk to North America's forests. Foreign vessels entering Canadian and U.S. ports are regularly inspected for Lymantria dispar (Linnaeus) and for the presence of other potentially invasive insects, including suspicious lasiocampid eggs. However, eggs are difficult to identify based on morphological features alone. Here, we report on the development of two TaqMan (Roche Molecular Systems, Inc., Rotkreuz, Switzerland) assays designed to assist regulatory agencies in their identification of these insects. Developed using the barcode region of the cytochrome c oxidase I (COI) gene and run in triplex format, the first assay can detect Dendrolimus and Malacosoma DNA, and can distinguish North American from Eurasian Malacosoma species. The second assay is based on markers identified within the internal transcribed spacer 2 (ITS2) region and was designed to specifically identify D. sibiricus, while discriminating closely related Dendrolimus taxa. In addition to providing direct species identification in the context of its use in North America, the D. sibiricus assay should prove useful for monitoring the spread of this pest in Eurasia, where its range overlaps with those of the morphologically identical D. superans (Butler) and similar D. pini (Linnaeus). The assays described here can be performed either in the lab on a benchtop instrument, or on-site using a portable machine.

Phenological Features of the Spongy Moth, Lymantria dispar (L.) (Lepidoptera: Erebidae), in the Northernmost Portions of Its Eurasian Range.

The spongy moth, Lymatria dispar, is a classic example of an invasive pest accidentally introduced from Europe to North America, where it has become one of the most serious forest defoliators, as in its native range. The present study was aimed at (i) identifying the current northern limit of L. dispar's Eurasian range and exploring its northward expansion in Canada using pheromone trap data, and (ii) comparing northern Eurasian populations with those from central and southern regions with respect to male flight phenology, the sums of effective temperatures (SETs) above the 7 °C threshold necessary for development to the adult stage, and heat availability. We show that the range of L. dispar in Eurasia now reaches the 61st parallel, and comparisons with historical data identify the average speed of spread as 50 km/year. We also document the northern progression of L. dispar in southern Canada, where the actual northern boundary of its range remains to be identified. We show that the median date of male flight does not vary greatly between northern and southern regions of the spongy moth range in Eurasia despite climate differences. Synchronization of flight at different latitudes of the range is associated with an acceleration of larval development in northern Eurasian populations. Similar changes in developmental rate along a latitudinal gradient have not been documented for North American populations. Thus, we argue that this feature of spongy moths from northern Eurasia poses a significant invasive threat to North America in terms of enhanced risks for rapid northward range expansion.

A New Cypovirus-1 Strain as a Promising Agent for Lepidopteran Pest Control.

Now more than ever researchers provide more and more evidence that it is necessary to develop an ecologically friendly approach to pest control. This is reflected in a sharp increase in the value of the biological insecticide market in recent decades. In our study, we found a virus strain belonging to the genus Cypovirus (Reoviridae); the strain was isolated from Dendrolimus sibiricus, possessing attractive features as a candidate for mass production of biological agents for lepidopteran-pest control. We describe the morphological, molecular, and ecological features of the new Cypovirus strain. This strain was found to be highly virulent to D. sibiricus (the half-lethal dose is 25 occlusion bodies per second-instar larva) and to have a relatively wide host range (infecting representatives of five families of Lepidoptera: Erebidae, Sphingidae, Pieridae, Noctuidae, and Lasiocampidae). The virus strain showed a strong interaction with a nontoxic adjuvant (optical brightener), which decreased the lethal dose for both main and alternative hosts, decreased lethal time, and may expand the host range. Moreover, we demonstrated that the insecticidal features were preserved after passaging through the most economically suitable host. By providing strong arguments for the possible use of this strain in pest control, we call on virologists, pest control specialists, and molecular biologists to give more attention to the Cypovirus genus, which may lead to new insights in the field of pest control research and may provide significant advantages to compare with baculoviruses and Bacillus thuringiensis products which are nowadays main source of bioinsecticides. IMPORTANCE In this article, we describe a newly discovered cypovirus strain that displays features ideally suited for the development of a modern biological insecticide: high potency, relatively broad host range, true regulating effect, flexible production (possibility to choose host species for production), interaction with enhancing adjuvants, and ecologically friendly. Based on an alignment of CPV genomes, we suggest that the enhanced host range of this new strain is the sequence of evolutionary events that occurred after coinfections involving different CPV species within the same host. These findings suggest that we need to positively reconsider CPVs as prospective agents as biocontrol products.