FMRfamide-like immunoreactivity in the ventral nerve cord of the larval eastern spruce budworm,Choristoneura fumiferana (clemens) (Lepidoptera : Tortricidae)

Distribution of FMRFamide-like immunoreactivity was examined in the larval ventral nerve cord of the eastern spruce budworm, Choristoneura fumiferana (Lepidoptera : Tortricidae). Indirect immunofluorescent methods revealed the existence of 3 groups of FLI neurons in each ganglion. The neurons are distributed in a bilaterally symmetrical fashion at the anterodorsal, midlateral and posteroventral regions of the ganglia. There are 4 FMRFamide-like immunoreactive fiber tracts on the dorsal surface of the ganglia to which the anterodorsal FLI neurons project ipsilaterally, while the midlateral pair projects both ipsi-, and contralaterally. The last abdominal ganglion (AG8) has 4 additional pairs of FLI neurons; and axons from some of these extend into the median abdominal nerve, which suggests some role for this neuropeptide in the control of posterior structures of the larva.     

Chemoreception of sucrose and amino acids in second and fourth instars of the spruce budworm Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae)

We examined the responses of some gustatory neurons in various contact-chemoreceptor sensilla of second-instar larvae of the spruce budworm. These included the L1 and L2 sensilla on the maxillary palp, and the LST and MST sensilla on the galea. Our objective was to determine whether there were differences in the physiological characteristics of individual neurons between the early and late larval instars. Changes were observed in both some sugar-sensitive and amino acid-sensitive neurons. We also confirmed the presence of a water-sensitive neuron in the L2 sensillum. Our findings are discussed in relation to changes that occur during the development of both the host plant and the insect. To our knowledge, this is the first paper to examine the responses from contact-chemoreceptor sensilla of very young second-instar caterpillar larvae.     

Electrophysiological responses to sucrose from a gustatory sensillum on the larval maxillary palp of the spruce budworm,Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae)

The tip of the maxillary palp in the spruce budworm and other lepidoptera larvae contains a cluster of eight basiconic sensilla. Except for a few electrophysiological recordings from the entire group of these sensilla, no information is available on the response characteristics of any individual one. Using a compound microscope at 320x magnification, with a long working-distance objective, it has been possible to view individual sensilla and to record electrical responses from three of these to some gustatory stimuli in preliminary tests. One of them, sensillum (L1), contains a sugar-sensitive neuron whose responses to a range of sucrose concentrations are reported here. The physiological characteristics of this neuron differ from those of the known sugar-sensitive neuron in the lateral styloconic sensillum on the galea of these same insects. Input of chemosensory information about the same gustatory stimulus by more than one neuron in different mouthpart sensilla is discussed in relation to the feeding habits of the spruce budworm.     

Oviposition behavior in spruce budwormChoristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae)

Epicuticular waxes of balsam fir, Abies balsamea,act as oviposition stimulants for fertilized females of the eastern spruce budworm, Choristoneura fumiferana(Clem.). Those same waxes do not stimulate unfertilized females, nor do the latter respond to nonchemical cues such as leaf texture, shape, etc. Ablation experiments suggest that the chemoreceptors responsible for the perception of waxes are located on the proboscis.     

Physiological differences between two sugar-sensitive neurons in the galea and the maxillary palp of the spruce budworm larva Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae)

The L1 contact-chemoreceptor sensillum on the maxillary palp of the spruce budworm larva Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae) was examined electrophysiologically for its responses to stimulation by various pyranose and furanose sugars. The results were compared to those from previous work on the sugar-sensitive neuron of the LST sensillum on the galea. We show that the L1 contact-chemoreceptor sensillum contains one sugar-sensitive neuron with furanose but no pyranose sites. It has response characteristics that differ from those of the sugar-sensitive neuron in the LST. Behavioural 2-choice feeding experiments show that, even with both known sugar-sensitive neurons disabled, larvae can still discriminate between disks treated with either distilled water or alpha-D-glucose. We conclude that the epipharyngeal sensilla must thus also contain a sugar-sensitive neuron.     

Pattern analysis of eastern spruce budworm Choristoneura fumiferana dispersal

Dispersal has been proposed as an important mechanism in the broad‐scale synchronisation of insect outbreaks by linking spatially disjunct populations. Evidence suggests that dispersal is influenced by landscape structure, phenology, temperature, and air currents; however, the details remain unclear due to the difficulty of quantifying dispersal. In this study, we used data on the abundance and distribution of spruce budworm Choristoneura fumiferana larvae (potential dispersers) and adult male moths (dispersers) to make inference on the effects of air currents and host‐species abundance on dispersal. Hierarchical‐Bayesian and inverse modeling was used to explore 4 dispersal models: 1) isotropic dispersal; 2) directional‐dispersal; 3) directional‐and‐host‐species dispersal; and 4) host‐species dispersal. Despite their strong dependence on balsam fir Abies balsamea and spruce species Picea spp., the mapped basal area of these host species did not influence the pattern of dispersed moths. The model that best fit the data was the directional‐dispersal model, which showed that the prevailing dispersal direction was from the northwest (328°). We infer that the strong pattern of directional dispersal was due to a prevailing wind from the same direction. Our interpretation was corroborated by independent wind data during the period of active adult male budworm flight, particularly in the region with high larval abundance. Our results indicate that there was a relatively high probability of individuals flying at least 48 km with the wind where larvae abundance at source locations was also high. Such findings emphasize the importance of long‐distance dispersal on spatial distribution of adult male spruce budworms. Insight into the population‐level consequences of such dispersal patterns requires additional research.     

Occurrence of Cystosporogenes sp. (Protozoa, Microsporidia) in a multi-species insect production facility and its elimination from a colony of the eastern spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae)

We have isolated a microsporidium from a laboratory colony of the eastern spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae). Light and electron microscopic investigations showed that gross pathology and ultrastructure of our isolate are similar to those described for Cystosporogenes legeri from the European grape vine moth, Lobesia botrana. Comparative phylogenetic analysis of the small subunit rDNA using maximum likelihood, maximum parsimony, and neighbour joining distance methods revealed perfect homology with the C. legeri sequence. The microsporidian was infectious to other Choristoneura species, as well as Malacosoma disstria, Lymantria dispar, and Lambdina fiscellaria. Incubation of infected egg masses at 41 degrees C for 20 min followed by 30 min in 33% formaldehyde did not reduce disease incidence in larval offspring. Exposure of one or two generations to fumagillin at 6000 ppm or higher eliminated infection in adult moths, but also reduced colony fitness. A clean colony was established by conducting individual matings and selecting disease-free offspring.     

Measurement, analysis, and interpretation of mortality factors in insect survivorship studies, with reference to the spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae)

A theory and practical method are developed to evaluate insect survivorship as a continuous process in time and the effects of major factors that determine it, with a view to providing solutions to certain problems in survivorship studies. There are two ways of assessing mortality rates: nominal and marginal. A nominal rate is based on the number tallied in terms of the immediate cause of death; it depends on the method of observation employed and can be inconsistent between studies. A marginal rate is free of such inconsistency. The marginal rate attributable to the union of parasitism and disease is evaluated by frequent sampling in the field and rearing of each batch of samples in the laboratory for a short period of time. Comparing this rate with the field mortality rate, directly evaluated by sampling, determines marginal predation rate. The marginal parasitism and disease rates cannot be determined by observation. A computational method is developed to evaluate their approximate values. A step-by-step procedure of numerical calculations with data is provided in an appendix. The accuracy of estimating mortality depends on the frequency of sampling in the field and that of inspecting dead individuals in rearing. How frequently sampling and inspection should be conducted is discussed. The method of assessing parasitism developed here has a great advantage over the conventional method based on dissecting the samples, or rearing them through, that often results in a serious underestimation. Also discussed is the relative importance of mortality factors from the pest control point of view, and the concept of “joint contribution” is introduced. Other practical considerations include transportation of samples; rearing conditions; and how to deal with the “dead-in-the-field” in the samples, the “rearing-death syndrome,” and the “virtually dead” individuals. Received: August 25, 2000 / Accepted: May 17, 2001     

Electrophysiological investigations on tarsal chemoreceptors of the spruce budworm, Choristoneura fumiferana (Lepidoptera)

Electrophysiological investigations show that chemosensitive cells in tarsal setae of the spruce budworm are sensitive to at least two salts and possibly water. Two cells are involved in the salt response and one in the possible water response. Each seta also has a cell which responds to mechanical deformation. Stimulation with four sugars did not show that a specific sugar sensitive cell is present in these setae. Results are compared with those from other arthropods.     

Utility of microsatellites and mitochondrial DNA for species delimitation in the spruce budworm (Choristoneura fumiferana) species complex (Lepidoptera: Tortricidae)

Species identifications have been historically difficult in the Choristoneura fumiferana group, an important insect pest complex. We examined the utility of simple sequence repeats (SSRs, also referred to as microsatellites) and mitochondrial DNA (mtDNA) for delimiting and identifying eight currently recognized species sampled across North America. Four of these species formed discrete clusters using SSRs, while only two species were delimited with mtDNA. There was evidence for hybridization or incomplete lineage sorting between several species pairs. An integrative approach, using both phenotypic traits and molecular markers, allowed for the discrimination of more biologically relevant species units than did the use of molecular markers alone. As species are currently identified using putatively adaptive phenotypic traits, the differences observed between recognized species and neutral SSRs or mtDNA suggests that these species (or evolutionary significant units) have diverged via natural selection in spite of some gene flow.     

Not just for the birds: Spiders as natural enemies of spruce budworm (Choristoneura fumiferana,Clem.)

(1) The eastern spruce budworm (Choristoneura fumiferana, Clem.) is a native irruptive forest pest that defoliates spruce-fir forests throughout North America's boreal zone. (2) Past studies suggest that successful spruce budworm population control requires high natural mortality from a variety of sources, including predators, especially from parasitoids and birds. While well represented in many different ecosystems, the role of generalist predatory spiders in these boreal systems remains largely unstudied. (3) To determine the identity and percentage of spiders preying upon spruce budworm, we hand-collected spiders from balsam fir (Abies balsamea) in stands with relatively high spruce budworm densities from forests in insular Newfoundland and Labrador, Canada. (4) Using a spruce budworm-specific TaqMan real-time PCR assay, we successfully amplified spruce budworm DNA in 32% of collected spiders. After spider molecular barcoding, we found the web builders Grammonota angusta Dondale, Pityohyphantes (aff. subarcticus), Dictyna brevitarsa Emerton and Estrandia grandaeva (Keyserling) represented 58% of the spiders feeding on spruce budworm, and the wandering hunter Philodromus rufus Dondale represented 11.8%. (5) Our molecular approach was an effective means with which to identify recently consumed prey and natural enemies in this boreal system.     

Integrating morphology and mitochondrial DNA for species delimitation within the spruce budworm (Choristoneura fumiferana) cryptic species complex (Lepidoptera: Tortricidae)

Species in cryptic complexes tend to be very difficult, if not impossible, to identify using morphological characters. One such complex is the spruce budworm (Choristoneura fumiferana Clemens, 1865) species group, an economically important group of Nearctic forest pests. Morphological, ecological, behavioural and genetic characters have been studied to try to understand the taxonomy of this group better, but diagnostic character states differ in frequency rather than being complete replacements between each species. We used mitochondrial DNA (mtDNA), together with a new morphology‐based character system that focuses on forewing colour components, to determine if one or a combination of character sources can be used for species diagnoses within the spruce budworm complex. We characterized 47 forewing morphometric measurements and sequenced a 470 bp region of cytochrome c oxidase I mtDNA for 111 ingroup individuals comprising five taxa within the complex. Larval host association and coloration or adult pheromone attraction were used as the prior method for grouping individuals. Our results showed that linear discriminant analysis of morphometric wing characters gave unique clusters for all species on the first and second canonical axes, except for a partial overlap between C. fumiferana and C. biennis, which are not sympatric in nature. In contrast, mtDNA distinguished C. fumiferana, C. pinus pinus Freeman, 1953 and a group of western species, but the three western species (C. occidentalis Freeman, 1967 , C. biennis Freeman, 1967 and C. lambertiana Busck, 1915) shared mtDNA haplotypes. On the basis of the linear discriminant analysis of the combined character set, this study supports the application of both morphology and mtDNA within a framework of integrative taxonomy as the most accurate method for species identification. Furthermore, it demonstrates the utility of quantitative colour analysis, which may be particularly helpful for groups in which colour characters are difficult to divide into discrete units due to intergrading hues.     

Vertical transmission of Nosema fumiferanae (Microsporidia: Nosematidae) and consequences for distribution, post-diapause emergence and dispersal of second-instar larvae of the spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae)

We examined vertical transmission of Nosema fumiferanae in the eastern spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae), and how it affects overwintering distribution and survival and spring emergence and dispersal of second-instar larvae in outbreak populations. Females containing 5.0 x 10(5) spores or more consistently produced 100% infected progeny. Transmission efficiency was still 50% at burdens as low as 0.2 x 10(5) spores per moth. Infection intensity in offspring increased with maternal spore load but became highly variable above 25 x 10(5) spores per female. Nosema multiplied in second instars for at least 1 month after they entered dormancy, regardless of temperature (2 degrees C versus 21 degrees C). Infection did not affect the distribution of overwintering larvae in a white spruce canopy. Dormancy survival between late-summer and the following spring was lower in families from infected females and was negatively correlated with larval infection intensity. Infection delayed larval emergence from hibernacula in the spring and resulted in delayed dispersal of emerged larvae, at least when parasite prevalence and infection intensities were high. Infected larvae were less successful in establishing feeding sites after dispersal. Our results underscore the potential of Nosema infection to negatively affect processes early in the budworm life cycle.     

Physiology of a sucrose-sensitive cell on the galea of the eastern spruce budworm larva, Choristoneura fumiferana

ABSTRACT The lateral styloconic sensillum on the galea of the eastern spruce budworm larva Choristoneura fumiferana Clem. (Lepidoptera: Tortricidae) contains a cell which responds to sucrose. The electro-physiologial threshold for response is below 0.5 mM sucrose. The K_b for the response is 1.5 mM, and the V_max is 200 impulses/s. The physiological data are interpreted with respect to the sucrose-stimulated feeding behaviour of the larva.