Biodegradation of Pine Processionary Caterpillar Silk Is Mediated by Elastase- and Subtilisin-like Proteases.

Pine processionary caterpillar nests are made from raw silk. Fibroin protein is the main component of silk which, in the case of pine processionary caterpillar, has some unusual properties such as a higher resistance to chemical hydrolysis. Isolation of microorganisms naturally present in silk nests led to identification of Bacillus licheniformis and Pseudomonas aeruginosa strains that in a defined minimal medium were able to carry out extensive silk biodegradation. A LasB elastase-like protein from P. aeruginosa was shown to be involved in silk biodegradation. A recombinant form of this protein expressed in Escherichia coli and purified by affinity chromatography was able to efficiently degrade silk in an in vitro assay. However, silk biodegradation by B. licheniformis strain was mediated by a SubC subtilisin-like protease. Homologous expression of a subtilisin Carlsberg encoding gene (subC) allowed faster degradation compared to the biodegradation kinetics of a wildtype B. licheniformis strain. This work led to the identification of new enzymes involved in biodegradation of silk materials, a finding which could lead to possible applications for controlling this pest and perhaps have importance from sanitary and biotechnological points of view.

When insect pests build their own thermal niche: The hot nest of the pine processionary moth.

Temperature strongly drives physiological and ecological processes in ectotherms. While many species rely on behavioural thermoregulation to avoid thermal extremes, others build structures (nests) that confer a shelter against climate variability and extremes. However, the microclimate inside nests remains unknown for most insects. We investigated the thermal environment inside the nest of a temperate winter-developing insect species, the pine processionary moth (PPM), Thaumetopoea pityocampa. Gregarious larvae collectively build a silken nest at the beginning of the cold season. We tested the hypothesis that it provides a warmer microenvironment to larvae. First, we monitored temperature inside different types of nests varying in the number of larvae inside. Overall, nest temperature was positively correlated to global radiation and air temperature. At noon, when global radiation was maximal, nest temperature exceeded air temperature by up to 11.2-16.5 °C depending on nest type. In addition, thermal gradients of amplitude from 6.85 to 15.5 °C were observed within nests, the upper part being the warmest. Second, we developed a biophysical model to predict temperature inside PPM nests based on heat transfer equations and to explain this important temperature excess. A simple model version accurately predicted experimental measurements, confirming that nest temperature is driven mainly by radiation load. Finally, the model showed that nest temperature increases at the same rate as air temperature change. We conclude that some pest insects already live in warm microclimates by building their own sheltering nest. This effect should be considered when studying the impact of climate change on phenology and distribution.

Shifted phenology in the pine processionary moth affects the outcome of tree-insect interaction.

In the Mediterranean and temperate regions, an increase in the frequency and intensity of drought events has been recorded, probably due to climate change. In consequence, trees will more frequently experience hydric stress, a condition that can be expected to affect insect-tree interactions, while adaptation mechanisms may be further in course. The effect of tree water stress on the performance of two allochronic populations of Thaumetopoea pityocampa was here studied. Namely, we compared a unique population of this insect, in which the larvae develop in the summer (SP), with the typical population having winter larval development (WP), to test the adaptation hypothesis to host plant status. Larvae of each population were fed on needles of young potted Pinus pinaster plants under two water supply regimes: (i) well-watered (control) and (ii) subjected to 3 months of drought stress. Compared to control, stressed plants had higher amounts of soluble sugars, phenols, and higher C/N ratio, whereas water content and chlorophylls concentrations were lower. In general, T. pityocampa larvae had lower performances on water-stressed plants, as shown by lower survival rates, lower needle consumption, and longer development times. Yet, the detrimental effects of tree stress were only significant for the WP larvae, while SP larvae were able to overcome such conditions. Results demonstrate that tree water stress can negatively affect T. pityocampa populations. Furthermore, the evidence is also provided that responses to the physiological condition of the host trees may occur at the population level, as a result of adaptation mechanisms driven by climate change.

Deciphering the demographic history of allochronic differentiation in the pine processionary moth Thaumetopoea pityocampa.

Understanding the processes of adaptive divergence, which may ultimately lead to speciation, is a major question in evolutionary biology. Allochronic differentiation refers to a particular situation where gene flow is primarily impeded by temporal isolation between early and late reproducers. This process has been suggested to occur in a large array of organisms, even though it is still overlooked in the literature. We here focused on a well-documented case of incipient allochronic speciation in the winter pine processionary moth Thaumetopoea pityocampa. This species typically reproduces in summer and larval development occurs throughout autumn and winter. A unique, phenologically shifted population (SP) was discovered in 1997 in Portugal. It was proved to be strongly differentiated from the sympatric "winter population" (WP), but its evolutionary history could only now be explored. We took advantage of the recent assembly of a draft genome and of the development of pan-genomic RAD-seq markers to decipher the demographic history of the differentiating populations and develop genome scans of adaptive differentiation. We showed that the SP diverged relatively recently, that is, few hundred years ago, and went through two successive bottlenecks followed by population size expansions, while the sympatric WP is currently experiencing a population decline. We identified outlier SNPs that were mapped onto the genome, but none were associated with the phenological shift or with subsequent adaptations. The strong genetic drift that occurred along the SP lineage certainly challenged our capacity to reveal functionally important loci.

Influence of Host Plant on Thaumetopoea pityocampa Gut Bacterial Community.

Microbial communities associated to the gut of insects are attracting an increasing interest, mainly because of their role in influencing several host life-traits. The characterization of the gut microbial community is pivotal for understanding insect ecology and, thus, to develop novel pest management strategies. The pine processionary moth, Thaumetopoea pytiocampa (Denis & Schiff.) (Lepidoptera: Thaumetopoeidae), is a severe defoliator of pine forests, able to feed on several pine species. In this work, we performed a metabarcoding analysis to investigate, for the first time, the diversity of the gut bacterial community of pine processionary larvae associated with three different host pine species (Pinus halepensis, Pinus nigra subsp. laricio, and Pinus pinaster). We found that the gut microbial community of T. pityocampa larvae collected on P. halapensis was different from that associated with larvae collected from P. nigra and P. pinaster. Moreover, the high presence of bacteria belonging to the genera Modestobacter, Delftia, and unidentified Methylobacteriaceae retrieved in larvae feeding on P. halapensis suggested that specific interactions can occur. Our results provide the evidence that different host plant differently impact on the microbiota diversity of T. pityocampa larvae, contributing to the general knowledge of this pest with information that could be useful in shaping the next generation of pest control strategies.

Host range expansion is density dependent.

The realized host range of herbivores is expected to increase with herbivore population density. Theory also predicts that trait similarity and phylogenetic relatedness between native and exotic plants is expected to increase the susceptibility of introduced plants to feeding by native herbivores. Whether the ability of native herbivores to extend their host range to introduced species is density dependent is still unknown. We addressed this question by monitoring pine processionary moth (PPM, Thaumetopoea pityocampa) attacks during nine consecutive years on 41 pine species (8 native and 33 introduced) planted in an arboretum. The survey encompassed latent and outbreak periods. A total of 28 pine species were attacked by PPM. There was no difference in the probability of attack between native and introduced pine species. Host range increased and was more phylogenetically clustered during outbreak than latent periods. When population density increased, PPM expanded its diet breadth by attacking introduced pine species that were closely related to native hosts. This study demonstrates the density dependence of host range expansion in a common pine herbivore. Importantly, it supports the idea that the degree of phylogenetic proximity between host species can be a better predictor of attacks than the introduction status, which may help to predict the outcomes of new plant-herbivore interactions.

Large Male Caterpillars Are the Primary Builders: Exploring Tent Construction and Foraging Behaviour in Gregarious Pine Processionary Caterpillar.

As a social organism, living in a communal structure is one of the most important physical barriers against environmental elements and natural enemies. Thaumetopoea pityocampa (Notodontidae, Thaumetopoeinae) caterpillars are conifer pests that spend most of their larval stage in winter. Although T. pityocampa holds economic and medical significance, the tent construction and foraging behaviour are poorly understood. We observed the tent construction behaviour in autumn (October and November) when third- and fourth-instar T. pityocampa caterpillars build the 'winter tent' that can withstand winter conditions. Just before sunset, with no rain and temperatures over 12 °C, tent construction was undertaken by early active individuals, primarily larger male caterpillars. Early active caterpillars emerge from the tent first and spin silk on the tent for expansion and strength. Once temperatures dropped below 12 °C and twilight had passed, the early active caterpillars went out to forage and were later joined by the late active caterpillars, which were predominantly smaller females that had remained inside the tent. Foraging behaviour was continuously monitored for the first to fourth larval instars in the field. Foraging was more frequent in younger instars when environmental temperatures were warmer and became continuous and prolonged in later instar caterpillars as temperatures dropped. The final tent structure built by later instar caterpillars had the thickest layer of silk on the southern side of the tent compared to other orientations to receive maximum solar radiation during the winter. Our study provided additional insights into the collective nest building, foraging and social behaviours observed in Lepidoptera, as well as the roles of individuals within non-eusocial insect colonies.

Sequence, assembly and count datasets of viruses associated to the pine processionary moth Thaumetopoea pityocampa (Denis & Schiffermüller) (Lepidoptera, Notodontidae) identified from transcriptomic high-throughput sequencing.

The pine processionary moth Thaumetopoea pityocampa is a Lepidopteran pest species occurring in the Western Mediterranean. It causes heavy pine defoliations and it is a public and animal health concern because of its urticating caterpillars. Very little is known about the viruses associated to this species, as only two viruses were described so far. We here present a dataset corresponding to 34 viral transcripts, among which 27 could be confidently assigned to 9 RNA and DNA viral families (Iflaviridae, Reoviridae, Partitiviridae, Permutotetraviridae, Flaviviridae, Rhabdoviridae, Parvoviridae, Baculoviridae and PolyDNAviridae). These transcripts were identified from an original transcriptome assembled for the insect host, using both blast search and phylogenetic approaches. The data were acquired from 2 populations in Portugal and 2 populations in Italy. The transcripts were de novo assembled and used to identify viral sequences by homology searches. We also provide information about the populations and life stages in which each virus was identified. The data produced will allow to enrich the virus taxonomy in Lepidopteran hosts, and to develop PCR-based diagnostic tools to screen colonies across the range and determine the distribution and prevalence of the identified viral species.

Fungal Keratitis, or Misled by a Small Insect?

The pine processionary caterpillar is an insect that has multiple small, thin hairs around its body as a defense mechanism. These hairs have a hazardous effect on ocular structures and cause a broad range of reactions from conjunctivitis to endophthalmitis, referred to as ophthalmia nodosa. The diagnosis of the disease is based on the patient's history and the detection of the hairs on ocular structures. In this report, we present a patient with ophthalmia nodosa misdiagnosed as fungal keratitis, and the actual diagnosis was made by in vivo confocal microscopy. We also would like to increase awareness among ophthalmologists about the disease which has a growing distribution area due to climate change.

Profiling Volatile Terpenoids from Calabrian Pine Stands Infested by the Pine Processionary Moth.

Terpenoids make up the biggest and most diversified class of chemical substances discovered in plants, encompassing over 40,000 individual compounds. In conifers, the production of terpenoids, either as oleoresin or emitted as volatile compounds, play an important role in the physical and chemical defence responses against pathogens and herbivores. In the present work, we examined, for the first time to the best of our knowledge, the terpenic defensive relations of Calabrian pine (Pinus nigra subsp. laricio (Poiret) Maire), facing the attack of the pine processionary moth (Thaumetopoea pityocampa (Denis and Schiffermüller, 1775)), brought about in the open on adult plant individuals growing at two distinct forest sites. Among the volatile terpenoids emitted from pine needles, bornyl acetate [(4,7,7-trimethyl-3-bicyclo[2.2.1]heptanyl) acetate] was the most frequently and selectively associated with the infestation, increasing during the period of most intense trophic activity of the caterpillars (defoliation), and decreasing thereafter. Although further work is needed to clarify whether the observed response reflects defence reactions and/or they are involved in communication among the infested plants and their biotic environment, the present results boost the currently growing interest in the isolation and characterization of plant secondary metabolites that can be used to control pests, pathogens, and weeds.

New Data on the Range Expansion of the Thaumetopoea pityocampa (Lepidoptera: Notodontidae) 'ENA clade' in Greece: The Role of Bacterial Endosymbionts.

The pine processionary moth, Thaumetopoea pityocampa (Denis and Schiffermüller), is an important insect in the Mediterranean region, as it defoliates pines and its urticating hairs can cause allergic reactions in humans and animals. Moreover, this species exhibits an interesting genetic structure as recently a distinct East-North African mtDNA lineage ('ENA clade') has been described. This clade has been recently detected in Greek populations where it has currently expanded its range by replacing the 'endemic' T. pityocampa lineages. Here, we report new data on the rapid spread of 'ENA clade' in the Greek island Evoia in only a few years. As the underlying mechanisms of the 'ENA clade' range expansion has not been studied so far, we screened T. pityocampa for an infection with the heritable bacterial endosymbionts Wolbachia (Bacteria: Anaplasmataceae), Cardinium (Bacteria: Bacteroidaceae), Rickettsia (Bacteria: Rickettsiaceae) and Spiroplasma (Bacteria: Spiroplasmataceae). These bacteria can manipulate the reproduction of infected hosts, something that could potentially explain the rapid spread of 'ENA clade' lineage. Therefore, we screened 28 individuals that exhibited T. pityocampa 'ENA clade' and 'endemic' T. pityocampa haplotypes from nine populations scattered all over Greece. None of them was infected with any of the four endosymbionts, suggesting that these bacteria do not cause reproductive manipulations in T. pityocampa lineages and, thus, other factors should be explored in future research efforts.

[Variation in Thaumetopoea pityocampa infestation rate of Aleppo pine: Effect on dendrometric parameters in the Djelfa region forests (Saharan Atlas, Algeria)]. / Variation du taux d'infestation par Thaumetopoea pityocampa du pin d'Alep : effet sur les paramètres dendrométriques dans les forêts de la région de Djelfa (Atlas saharien, Algérie).

The forests of the Saharan Atlas represent the southern edge of the natural range of the Aleppo pine (Pinus halepensis Mill.). These are exposed to climatic stress in relation to drought and attacks of the pine processionary moth (Thaumetopoea pityocampa Schiff., denoted by CP in the French text). The purpose of our work is to study the variation of the infestation rate by the pine processionary moth and its density with climatic factors and dendrometric parameters. The infestation rate (7.82%) by the pine processionary moth and its density (2.74±2.61nests/tree) were lower in our pine forests than in the northern ones. It appeared that the attacks of the moth increase mainly with the number of days of frost and heatwave. The dendrometric parameters were negatively correlated with the moth's density per tree. On the other hand, the latter was positively correlated with the defoliation rate. The results were discussed in a biogeographic framework in the light of the current knowledge of the distribution of the pine processionary moth in the Aleppo pine, in relation with the climatic conditions.

Pine processionary caterpillar Thaumetopoea pityocampa envenomation in 109 dogs: A retrospective study.

Contact with the caterpillars of the pine processionary moth (CPPM) Thaumetopoea pityocampa induces severe local allergic reactions. The purpose of this large-scale retrospective cohort-study was to describe the clinical manifestations and related risk factors of CPPM exposure. This cohort-study included 109 dogs between the years of 2000 and 2016. Tongue lesions ranging from oedema to severe necrosis were observed in 94/109 dogs (86%). The following systemic signs were observed in 60/109 dogs (55%): vomiting (52/109, 48%), dyspnoea (6/109, 5%), hypovolemia (4/109, 4%) and diarrhoea (2/109, 2%). Based on the time elapsed from CPPM contact to the first oral flushing, three groups were defined: <2 h (group 1, 37/105, 35%), 2 h-6 h (group 2, 39/105, 37%) and >6 h (group 3, 29/105, 28%). Tongue necrosis (TN) at admission was significantly more common in the dogs in group 3 than those in groups 1 and 2 (45% vs. 5% and 5% respectively, p = 0.0002). In addition, the development of TN during hospitalisation was significantly more common in the dogs in group 3 (65%) than in those in the other groups (21% in group 1, p = 0.02) and 31% in group 2, p = 0.001). The dogs in group 3 presented a 14.63-fold higher risk of TN at admission and a 3.78-fold higher risk of developing necrosis during hospitalisation compared with the other groups. The survival rate after exposure was 97%. Long-term follow-up data were available for 69/109 dogs (63%). Twenty-three dogs (37%) had persistent, definitive TN without major consequences on quality of life. Elapsed time between contact and first oral flushing appears to be a key determinant for the progression of necrotic lesions, and the best results were observed when flushing occurred within 6 h of contact. The prognosis of CPPM envenomation is excellent, with a short hospitalisation duration.

An exhaustive inventory of coniferous trees in an agricultural landscape.

BACKGROUND: Various species of forest trees are commonly used for ornamental purposes and are therefore frequently found in non-forest ecosystems. These trees constitute a significant component of the trees outside forests (TOF). Although increasingly recognized as prominent feature of agricultural lands and built-up areas, not much is known, however, about TOF since they are generally absent from forest inventories. NEW INFORMATION: In the present study, we focus on the coniferous tree species that constitute potential hosts for a forest defoliator, the pine processionary moth Thaumetopoeapityocampa Den. & Schiff. (Lepidoptera, Notodontidae). We carried out an exhaustive inventory of all pines (Pinus spp.), cedars (Cedrus spp.) and Douglas-fir (Pseudotsugamenziesii) in a 22 × 22 km study window located in the open-field region of Beauce in the centre of France. We recorded a total of 3834 individuals or small groups host trees corresponding a density of 7.9 occurrences per 100 ha. We provide the spatial coordinates of the points without differentiation between tree species.

Comparative analysis of two phenologically divergent populations of the pine processionary moth (Thaumetopoea pityocampa) by de novo transcriptome sequencing.

The pine processionary moth Thaumetopoea pityocampa is a Mediterranean lepidopteran defoliator that experiences a rapid range expansion towards higher latitudes and altitudes due to the current climate warming. Its phenology - the time of sexual reproduction - is certainly a key trait for the local adaptation of the processionary moth to climatic conditions. Moreover, an exceptional case of allochronic differentiation was discovered ca. 15 years ago in this species. A population with a shifted phenology (the summer population, SP) co-exists near Leiria, Portugal, with a population following the classical cycle (the winter population, WP). The existence of this population is an outstanding opportunity to decipher the genetic bases of phenology. No genomic resources were so far available for T. pityocampa. We developed a high-throughput sequencing approach to build a first reference transcriptome, and to proceed with comparative analyses of the sympatric SP and WP. We pooled RNA extracted from whole individuals of various developmental stages, and performed a transcriptome characterisation for both populations combining Roche 454-FLX and traditional Sanger data. The obtained sequences were clustered into ca. 12,000 transcripts corresponding to 9265 unigenes. The mean transcript coverage was 21.9 reads per bp. Almost 70% of the de novo assembled transcripts displayed significant similarity to previously published proteins and around 50% of the transcripts contained a full-length coding region. Comparative analyses of the population transcriptomes allowed to investigate genes specifically expressed in one of the studied populations only, and to identify the most divergent homologous SP/WP transcripts. The most divergent pairs of transcripts did not correspond to obvious phenology-related candidate genes, and 43% could not be functionally annotated. This study provides the first comprehensive genome-wide resource for the target species T. pityocampa. Many of the assembled genes are orthologs of published Lepidoptera genes, which allows carrying out gene-specific re-sequencing. Data mining has allowed the identification of SNP loci that will be useful for population genomic approaches and genome-wide scans of population differentiation to identify signatures of selection.

Are heat waves susceptible to mitigate the expansion of a species progressing with global warming?

A number of organisms, especially insects, are extending their range in response of the increasing trend of warmer temperatures. However, the effects of more frequent climatic anomalies on these species are not clearly known. The pine processionary moth, Thaumetopoea pityocampa, is a forest pest that is currently extending its geographical distribution in Europe in response to climate warming. However, its population density largely decreased in its northern expansion range (near Paris, France) the year following the 2003 heat wave. In this study, we tested whether the 2003 heat wave could have killed a large part of egg masses. First, the local heat wave intensity was determined. Then, an outdoor experiment was conducted to measure the deviation between the temperatures recorded by weather stations and those observed within sun-exposed egg masses. A second experiment was conducted under laboratory conditions to simulate heat wave conditions (with night/day temperatures of 20/32°C and 20/40°C compared to the control treatment 13/20°C) and measure the potential effects of this heat wave on egg masses. No effects were noticed on egg development. Then, larvae hatched from these egg masses were reared under mild conditions until the third instar and no delayed effects on the development of larvae were found. Instead of eggs, the 2003 heat wave had probably affected directly or indirectly the young larvae that were already hatched when it occurred. Our results suggest that the effects of extreme climatic anomalies occurring over narrow time windows are difficult to determine because they strongly depend on the life stage of the species exposed to these anomalies. However, these effects could potentially reduce or enhance the average warming effects. As extreme weather conditions are predicted to become more frequent in the future, it is necessary to disentangle the effects of the warming trend from the effects of climatic anomalies when predicting the response of a species to climate change.