Genotypic variation and presence of rare genotypes among Douglas-fir tussock moth multicapsid nucleopolyhedrovirus (OpMNPV) isolates in British Columbia

The Douglas-fir tussock moth (Orgyia pseudotsugata) multicapsid nucleopolyhedrovirus (OpMNPV) is periodically applied to suppress Douglas-fir tussock moth populations in British Columbia and in the western United States. The strain of OpMNPV in the product currently used for suppression is not genetically distinct from naturally occurring OpMNPV. To separate the mortality caused by the applied virus from that caused by the naturally occurring virus, a rare and genetically distinct strain of OpMNPV must be applied. To learn more about the genotypic diversity of OpMNPV populations in BC and to identify rare strains in this region, viral DNA was extracted from larvae reared from 208 field-collected egg masses found in five geographic regions of British Columbia and subjected to REN analysis. Nine, 12, and 9 different genotypes were detected using PstI, SalI, and HindIII, respectively. When the PstI, SalI, and HindIII profiles for each pure (single strain) isolate were grouped and considered as a combined PstI-SalI-HindIII genotype, 23 different genotypes were identified among 185 isolates. Nine rare OpMNPV genotypes were selected as ideal candidates for use as a potential 'marker strain' to accurately determine the efficacy of the treatment.     

Production and efficacy of baculviruses

Several baculvirusus of nuclear polyhedrosis virus (NPV) have been produced and tested for microbial control of various Lepidoptera spp. To date, there are three registered preparations of NPV that are exempt from the requirement of tolerance by the Environmental Protection Agency (EPA) in the United States (US). The first and only commercially available viral preparation used in agriculture was developed by Sandoz, Inc. under the name of Elcar® for control of Heliothis spp. on cotton. The other two baculovirus preparations were developed and registered by the US Department of Agriculture (USDA) for control of Douglas-fir tussock moth and gypsy moth on forests. Several methods are being used for production of NPV viruses: (1) field collection of diseased larvae, (2) laboratory rearing of insects followed by infection with viral inoculum, (3) tissue culture. and (4) tissue culture and mass rearing larvae. Recent progress in mass production of insect virus points toward the adoption of tissue culture with the whole organism technology for production of a standardized viral product. The practical usefulness of various baculovirus preparations has been demonstrated for protection of forests from defoliation by various lepidopterous species. In agriculture, Elcar® has been successfully marketed and has been very well received for use in integrated pest management on cotton. Recent development also demonstrated that use of adjuvants further increase the efficacy of Elcar® against Heliothis spp. on cotton.     

Inactivation of nuclear polyhedrosis virus (Baculovirus subgroup A) by monochromatic UV radiation.

Monochromatic radiation at wavelengths of 290, 300, 310, and 320 nm inactivated occluded nuclear polyhedrosis virus of the Douglas-fir tussock moth, Orgyia pseudotsugata (McDunnough). Data indicate that all of the wavelengths are capable of causing virus inactivation; much greater fluences are needed for virus inactivation as the wavelength increases.     

Inactivation of nuclear polyhedrosis virus (Baculovirus subgroup A) by monochromatic UV radiation

Monochromatic radiation at wavelengths of 290, 300, 310, and 320 nm inactivated occluded nuclear polyhedrosis virus of the Douglas-fir tussock moth, Orgyia pseudotsugata (McDunnough). Data indicate that all of the wavelengths are capable of causing virus inactivation; much greater fluences are needed for virus inactivation as the wavelength increases.     

Characterization of a nucleopolyhedrovirus from the vapourer moth, Orgyia antiqua (Lepidoptera Lymantriidae).

The first characterization of a nucleopolyhedrovirus (NPV Baculoviridae) isolated from the vapourer moth, Orgyia antiqua (Lepidoptera Lymatriidae), in the United Kingdom is presented. Transmission electron microscopy revealed that the virus nucleocapsid rods were singly enveloped in the polyhedron inclusion body (PIB) so that the virus is assigned to the SNPV subgenus of the Baculoviridae. Restriction endonuclease analyses of viral DNA indicated a genomic size of approximately 148 kb. Restriction profiles of O. antiqua SNPV closely resembled those of heterologous NPVs isolated from four different Orgyia species and was most similar to a North American SNPV isolate from the Douglas-fir tussock moth, O. pseudotsugata. In host range tests, O. antiqua SNPV was not infectious to 23 lepidopteran species representing four families. Heterologous Orgyia NPV isolates were permissive in O. antiqua larvae. Using a diet plug bioassay method, the median lethal dose response (LD(50)) for this virus in second and third instar O. antiqua larvae were estimated at 52 and 539 PIBs per larva, respectively.     

Thermal inactivation characteristics of two strains of nucleopolyhedrosis virus (Baculovirus subgroup A) pathogenic for Orgyia pseudotsugata

Two nucleopolyhedrosis viruses of the Douglas-fir tussock moth, Orgyia pseudotsugata, one with a single nucleocapsid per envelope (SV) and one with multiple nucleocapsids per envelope (BV), are inactivated by a first-order reaction at 55° and 60°C. BV is the more thermostable of the two viruses: At both test temperatures, it has a lower inactivation rate than SV. BV is also the more virulent of the two viruses, with respect to acute course of the disease and severity of the histological lesions. The greater thermostability of BV and the acute course of the disease caused by this pathogen support the choice of BV as the virus most suitable for industrial production and field use.     

How mathematical epidemiology became a field of biology: a commentary on Anderson and May (1981) ‘The population dynamics of microparasites and their invertebrate hosts’

We discuss the context, content and importance of the paper ‘The population dynamics of microparasites and their invertebrate hosts’, by R. M. Anderson and R. M. May, published in the Philosophical Transactions of the Royal Society as a stand-alone issue in 1981. We do this from the broader perspective of the study of infectious disease dynamics, rather than the specific perspective of the dynamics of insect pathogens. We argue that their 1981 paper fits seamlessly in the systematic study of infectious disease dynamics that was initiated by the authors in 1978, combining effective use of simple mathematical models, firmly rooted in biology, with observable or empirically measurable ingredients and quantities, and promoting extensive capacity building. This systematic approach, taking ecology and biology rather than applied mathematics as the motivation for advance, proved essential for the maturation of the field, and culminated in their landmark textbook of 1991. This commentary was written to celebrate the 350th anniversary of the journal Philosophical Transactions of the Royal Society.     

Effects of long-term storage on potency of TM Biocontrol-1, the registered viral insecticide of Orgyia pseudotsugata (Lepidoptera: Lymantriidae)

The article presents the results of bioassaying 39 samples of TM Biocontrol-1, a viral insecticide, from 10 different lots and various sizes of vacuum-sealed packages that were stored at -10 degrees C for 5-15 yr. This is the first study to present potency data for a registered virus product stored for this length of time. Laboratory bioassays in insects from the same colony from which the TM Biocontrol-1 was produced showed that the stored viral insecticide is still effective, although it lost approximately 30% of its effectiveness during storage. A direct correlation of this loss with the length of time in storage is not apparent. Bioassays also showed that there are significant differences in the susceptibility of Douglas-fir tussock moth, Orgyia pseudotsugata (McDunnough), larvae from different geographic regions to OpMNPV (family Baculoviridae, genus Nucleopolyhedrovirus) infection. Package size did not affect the potency of stored TM Biocontrol-1. There were no clear, significant differences in the effectiveness among lots of TM Biocontrol-1 processed by different organizations.     

Exploring stable pattern formation in models of tussock moth populations

1. The western tussock moth ( Orgyia vetusta ) at the University of California Bodega Marine Reserve (Sonoma County, California, USA) exhibits dense, localized populations in the midst of extensive habitats where variation in host plant quality or predator abundance is unable to explain the restricted extent of the outbreaks.
2. Two primary features suggest that the host patterning is intrinsically generated: (i) female tussock moths are wingless, producing a low effective dispersal distance for the hosts; and (ii) the tussock moth population is attacked by several species of widely dispersing wasp and fly parasitoids.
3. We consider a set of spatially explicit host–parasitoid models to examine whether intrinsically generated patterns are possible within this system. These models include a spatially extended Nicholson–Bailey model to examine general features of pattern formation in host–parasitoid systems, and two system-specific models, an individual-based simulation and a population-level analytic model, to examine the details of this empirical system.
4. Both stable patterning and rapid extinction of the host population are initial-condition dependent outcomes of the general and specific models, implying that an intrinsically generated stable host pattern is feasible within the tussock moth system.
5. Stable patterning is enhanced by a large parasitoid-to-host dispersal ratio, local host resource limitation, and increased parasitism at the host patch's edge.     

Development and Evaluation of Methods To Detect Nucleopolyhedroviruses in Larvae of the Douglas-Fir Tussock Moth, Orgyia pseudotsugata (McDunnough)

Various molecular methods are used to detect pathogenic microorganisms and viruses within their hosts, but these methods are rarely validated by direct comparison. Southern hybridization, enzyme-linked immunosorbent assay (ELISA), and a novel DNA extraction/PCR assay were used to detect Orgyia pseudotsugata multiple nucleopolyhedrovirus (OpMNPV) in Douglas-fir tussock moth larvae. PCR was more sensitive than Southern hybridization and ELISA at detecting semipurified virus. ELISA, however, was the most accurate method for detecting virus within larvae, given that Southern hybridization and PCR produced false-negative results (31% and 2.5%, respectively). ELISA may be preferable in some applications because virus infections can be quantified (r² = 0.995). These results may be applicable to both applied and academic research that seeks to accurately identify the incidence of viruses and microorganisms that regulate insect populations.     

Resources and dispersal as factors limiting a population of the tussock moth (Orgyia vetusta), a flightless defoliator

The western tussock moth (Orgyia vetusta) is very abundant on one stand of bush lupine (Lupinus arboreus) at the Bodega Marine Reserve (Sonoma Co, Calif., USA), but is sparse or absent on nearby stands. To determine what controls its abundance, both within the outbreak area and more globally, I performed experimental manipulations of resource availability and dispersal. To measure resource limitation, I inoculated 30 caged and 30 uncaged bushes with a realistic range of numbers of tussock moth eggs. On caged bushes, starvation led to density-dependent reductions in survival, pupal weight and fecundity. Larvae on uncaged bushes experienced density-independent ant predation on early instars and density-dependent emigration by late instars. From the results of this experiment, I predicted the density of a resource-limited tussock moth population. The predictions agreed fairly well with data from the outbreak area in 1992. To measure dispersal by the moth, which has flightless adult females, I released 30,000 tussock moth eggs at a central point in each of two uninfested lupine stands, and censused larvae weekly in a circle of radius 16 m until pupation. Median displacement over one entire generation was only 2 m, or about 2 bush radii. Rearing experiments indicated that bushes outside the out-break area are fully nutritionally suitable for the moth. I conclude that two major factors limiting the population are resources (within the outbreak area) and inefficient dispersal (more globally).     

Development and evaluation of methods to detect nucleopolyhedroviruses in larvae of the Douglas-fir tussock moth, Orgyia pseudotsugata (McDunnough)

Various molecular methods are used to detect pathogenic microorganisms and viruses within their hosts, but these methods are rarely validated by direct comparison. Southern hybridization, enzyme-linked immunosorbent assay (ELISA), and a novel DNA extraction/PCR assay were used to detect Orgyia pseudotsugata multiple nucleopolyhedrovirus (OpMNPV) in Douglas-fir tussock moth larvae. PCR was more sensitive than Southern hybridization and ELISA at detecting semipurified virus. ELISA, however, was the most accurate method for detecting virus within larvae, given that Southern hybridization and PCR produced false-negative results (31% and 2.5%, respectively). ELISA may be preferable in some applications because virus infections can be quantified (r(2) = 0.995). These results may be applicable to both applied and academic research that seeks to accurately identify the incidence of viruses and microorganisms that regulate insect populations.     

The coniferous tussock moth (<Emphasis Type="Italic">Calliteara abietis</Emphasis>, Lepidoptera,Lymantriidae) in Buryatia

The coniferous tussock moth Calliteara abietis Denis et Schiffermüller, 1775 (Lepidoptera, Lymantriidae) outbreaks occur in Eastern Siberia. The biological characteristics and outbreak dynamics of this species in the late twentieth century and in the twenty-first century are described. Outbreaks of the moth are shown to occur both separately and along with Siberian and white striped (in Sakhalin) lappet moths.     

Phoresy ofTelenomus sp. (Scelionidae: Hymenoptera), an egg parasitoid of the tussock mothEuproctis taiwana

The phoretic relationship between the egg parasitoidTelenomus sp. cf.euproctidis Wilcox and its host the tussock mothEuproctis taiwana was studied in Okinawa, Japan. One third of the female moths studied in the field carried female parasitoid adults. No male moths carried parasitoids. Parasitoids were observed only in the anal tuft of the moth. Laboratory observation revealed that most of the parasitoids left the body of the moth at the time of the first oviposition of their host and proceeded to lay eggs on the moth egg masses.     

应用性信息素迷向法防治茶毛虫的田间试验

2003年在贵州省黔南州都匀茶场进行了性信息素迷向法防治茶毛虫Euproctis pseudoconspersaStrand的田间试验。通过比较处理区和对照区内茶毛虫发蛾量、交配率和后代虫口密度,发现剂量为0.5mg的性信息素释放器按5 m的间隔施放的防治效果显著好于其它处理,它在越冬代和第1代试验中茶毛虫诱蛾量分别减少83.90%和70.25%,交配率分别下降78.18%和73.21%,虫口密度减少59.02%和45.71%。试验结果表明性信息素迷向法可以作为防治茶毛虫的一种手段。     

Density-dependent resistance of the gypsy moth <Emphasis Type="Italic">Lymantria dispar</Emphasis> to its nucleopolyhedrovirus,and the consequences for population dynamics

The processes controlling disease resistance can strongly influence the population dynamics of insect outbreaks. Evidence that disease resistance is density-dependent is accumulating, but the exact form of this relationship is highly variable from species to species. It has been hypothesized that insects experiencing high population densities might allocate more energy to disease resistance than those at lower densities, because they are more likely to encounter density-dependent pathogens. In contrast, the increased stress of high-density conditions might leave insects more vulnerable to disease. Both scenarios have been reported for various outbreak Lepidoptera in the literature. We tested the relationship between larval density and disease resistance with the gypsy moth (Lymantria dispar) and one of its most important density-dependent mortality factors, the nucleopolyhedrovirus (NPV) LdMNPV, in a series of bioassays. Larvae were reared in groups at different densities, fed the virus individually, and then reared individually to evaluate response to infection. In this system, resistance to the virus decreased with increasing larval density. Similarly, time to death was faster at high densities than at lower densities. Implications of density–resistance relationships for insect–pathogen population dynamics were explored in a mathematical model. In general, an inverse relationship between rearing density and disease resistance has a stabilizing effect on population dynamics.     

Unexpected spatial patterns in an insect outbreak match a predator diffusion model

An outbreak of the western tussock moth ( Orgyia vetusta) with a sharp spatial boundary has been hypothesized to be the result of an interaction with mobile parasitoids. We report here a striking correspondence between the qualitative prediction of a general model for a mobile predator and a sessile prey and initial observations of the outbreak of western tussock moth. The model makes the counterintuitive prediction that the prey species will have its highest population density at the edge of an outbreak, which corresponds to observations made of the western tussock moth. This is a striking pattern which should be looked for in other population systems.     

A note on the relationship between outbreaks of the oriental tussock moth,Euproctis flava and weather conditions

A relationship between outbreaks of the oriental tussock moth, Euproctis flava, in Akita Prefecture and weather conditions have been analyzed in the present paper. The results obtained are summarized as follows: Outbreaks were, in most cases, observed at intervals of 7–9 years, and the population fluctuation was considered to be regular, when small rises of population density were taken into consideration. The condition of low temperature and decreased percentage of sunny hours in spring and summer has a great influence upon the occurrence of outbreaks, and a big outbreak seems to come when a peak of the intrinsic population fluctuation occurs just simultaneously with such a weather condition.     

Phylogenetic analysis of <Emphasis Type="Italic">Orgyia pseudotsugata</Emphasis> single-nucleocapsid nucleopolyhedrovirus

The Douglas-fir tussock moth Orgyia pseudotsugata (Lepidoptera: Lymantriidae) is a frequent defoliator of Douglas-fir and true firs in western USA and Canada. A single nucleopolyhedrovirus (SNPV) isolated from O. pseudotsugata larvae in Canada (OpSNPV) was previously analyzed via its polyhedrin gene, but is phylogenetic status was ambiguous. Sequences of four conserved baculovirus genes, polyhedrin, lef-8, pif-2 and dpol, were amplified from OpSNPV DNA in polymerase chain reactions using degenerate primer sets and their sequences were analyzed phylogenetically. The analysis revealed that OpSNPV belongs to group II NPVs and is most closely related to SNPVs that infect O. ericae and O. anartoides, respectively. These results show the need for multiple, concatenated gene phylogenies to classify baculoviruses. Foundation item: Supported by a scholarship from the European Union (Functional Biodiversity and Crop Protection), contract n^o HPMT-CT-2000-00199.