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The Carpathians are a range of mountains forming an arc roughly 1,500 km long across Central and Eastern Europe. They are an important area for biodiversity and belong to one of the major refuges of the last ice ages for many organisms. The forests of the Carpathians are dominated by spruce, which have suffered continuous outbreaks of the eight spined spruce bark beetle, Ips typographus, in recent decades. The phylogeography of this spruce pest is well documented, however, little is known on small scale, i.e., the Carpathians. Here we applied a mitochondrial marker and studied the genetic variation and structure of Carpathian populations and compared data with published one from other European populations. Twelve haplotypes were characterized and 42 % of those were not detected in other European populations. Despite a slight genetic structure, differences were observed in the haplotype distribution and diversity between the Western/Southern Carpathians and the Eastern Carpathians reflecting at least two potential refugial areas for I. typographus within the Carpathian mountain system. Further data show that the Eastern Beskidian Mountains of the Carpathians could act as barrier for several European haplotypes. This small-scale analysis reveals that the Carpathians have been an important glacial and post-glacial refuge for I. typographus. This information is important for a preventive and reactive forest management. [ABSTRACT FROM AUTHOR]

The common endosymbiotic Wolbachia bacteria influence arthropod hosts in multiple ways. They are mostly recognized for their manipulations of host reproduction, yet, more recent studies demonstrate that Wolbachia also impact host behavior, metabolic pathways and immunity. Besides their biological and evolutionary roles, Wolbachia are new potential biological control agents for pest and vector management. Importantly, Wolbachia-based control strategies require controlled symbiont transfer between host species and predictable outcomes of novel Wolbachia-host associations. Theoretically, this artificial horizontal transfer could inflict genetic changes within transferred Wolbachia populations. This could be facilitated through de novo mutations in the novel recipient host or changes of haplotype frequencies of polymorphic Wolbachia populations when transferred from donor to recipient hosts. Here we show that Wolbachia resident in the European cherry fruit fly, Rhagoletis cerasi, exhibit ancestral and cryptic sequence polymorphism in three symbiont genes, which are exposed upon microinjection into the new hosts Drosophila simulans and Ceratitis capitata. Our analyses of Wolbachia in microinjected D. simulans over 150 generations after microinjection uncovered infections with multiple Wolbachia strains in trans-infected lines that had previously been typed as single infections. This confirms the persistence of low-titer Wolbachia strains in microinjection experiments that had previously escaped standard detection techniques. Our study demonstrates that infections by multiple Wolbachia strains can shift in prevalence after artificial host transfer driven by either stochastic or selective processes. Trans-infection of Wolbachia can claim fitness costs in new hosts and we speculate that these costs may have driven the shifts of Wolbachia strains that we saw in our model system. [ABSTRACT FROM AUTHOR]

The European cherry fruit fly Rhagoletis cerasi has been a field model for cytoplasmic incompatibility since the mid 1970s. Two Wolbachia strains were detected in this tephritid species and wCer2 was described as the CI inducing agent dividing European populations into two unidirectional incompatible groups, i.e. southern females produce viable offspring with northern males, whereas the reciprocal cross results in incompatibility. We detected three new Wolbachia strains by sequencing a multitude of plasmids derived from Wolbachia surface protein gene ( wsp) polymerase chain reaction (PCR) products. Strain-specific primers were developed allowing individual diagnosis without need for cloning. Hybridization of specific PCR products with a wsp oligonucleotide enhanced the detection limit significantly and revealed the presence of low-titre infections in some strains, in different ontogenetic stages and in adults of different age. We then performed a survey of strain prevalence and infection frequency in eight European regions. wCer1 was fixed in all populations, whereas wCer2 was detected only in the South. wCer3 frequency was the lowest without a clear distribution pattern. The abundance of wCer4 was homogenous across Europe. Like wCer2, wCer5 showed significant differences in spatial distribution. Our new findings of previously undetected and recombinant Wolbachia strains in R. cerasi reveal a major caveat to the research community not to overlook hidden Wolbachia diversity in field populations. Low-titres and geographical variability in Wolbachia diversity are expected to influence the outcome of Wolbachia population dynamics and Wolbachia-based insect population control and may create invasion barriers for expanding and artificially introduced Wolbachia strains. [ABSTRACT FROM AUTHOR]

Biological control is the purposeful introduction of parasites, predators, and pathogens to reduce or suppress pest populations. Wolbachia are inherited bacteria of arthropods that have recently attracted attention for their potential as new biocontrol agents. Wolbachia manipulate host reproduction by using several strate- gies, one of which is cytoplasmic incompatibility (Cl) [Stouthamer, R., Breeuwer, J. A. J. & Hurst G. D. D. (1999) Annu. Rev. Microbiol. 53, 71-1021. We established Wolbachia-infected lines of the medfly Ceratitis capitata using the infected cherry fruit fly Rhagoletis cerasi as donor. Wolbachia induced complete CI in the novel host. Laboratory cage populations were completely suppressed by single releases of infected males, suggesting that Wolbachia-induced Cl could be used as a novel environmentally friendly tool for the control of medfly populations. The results also encourage the introduction of Wolbachia into pest and vector species of economic and hygenic relevance to suppress or modify natural populations. [ABSTRACT FROM AUTHOR]

The Przewalski's horse (Equus caballus przewalskii) became extinct in the wild in the 1960s. Since 1997, captivebred horses have been released into the Gobi-B Strictly Protected Area (SPA) in southwestern Mongolia, and successful reproduction in the wild started in 1999. In 2002, the population formed 3 harem groups and 1 bachelor group (total 38) in the wild, and 3 harem groups (24) awaited release in summer 2003 within acclimatization enclosures, totaling 62 individuals. We used the stochastic population simulation model VORTEX to (1) identify key variables and their threshold values in population dynamics, (2) predict extinction risk, and (3) optimize project management and release regime by comparing model parameters with our population data. Maximum age of reproduction, foal mortality, and fecundity rates were key factors in population dynamics, while number of released animals, release interval, and duration of supplementation played lesser roles. The severity level of natural catastrophes had the greatest influence on extinction risk and population size according to VORTEX. Assuming a maximum reproductive age of 16 years, an initial population of>140 horses is necessary to achieve a 95% probability of survival over 100 years under the low-severity level of catastrophes scenario. The corresponding extinction risk for high-severity level of catastrophes is 37%, even for initial population sizes >500. The low natural growth rate of the Przewalsld's horse may have been the essential prerequisite for extinction in this remote area of Mongolia. However, uncertainty of results was high and limits the predictive value of the model. Comparisons between model parameters with observed population characteristics over the past 10 years reveal some discrepancies that may require readjustment of the model if present trends continue. While our model currently underestimates reproductive rate and foal survival, adult mortality tends to be higher than estimated in t... [ABSTRACT FROM AUTHOR]

Cytoplasmic incompatibility (CI) is induced in arthropods by the maternally inherited bacterium Wolbachia. When infected males mate with uninfected females or with females bearing a different Wolbachia variant, paternal chromosomes behave abnormally and embryos die. This pattern can be interpreted as resulting from two bacterial effects: One (usually termed rood, for modification) would affect sperm and induce embryo death, unless Wolbachia is also present in the egg, which implies the existence of a second effect, usually termed resc, for rescue. The fact that CI can occur in crosses between males and females infected by different Wolbachia shows that mod and resc interact in a specific manner. In other words, different compatibility types, or mod/resc pairs seem to have diverged from one (or a few) common ancestor(s). We are interested in the process allowing the evolution of modlresc pairs. Here this question is addressed experimentally after cytoplasmic injection into a single host species (Drosophila simulans) by investigating compatibility relationships between closely related Wolbachia variants naturally evolving in different dipteran hosts: D. simulans, Drosophila melanogaster, and Rhagoletis cerasi. Our results suggest that closely related bacteria can be totally or partially incompatible. The compatibility relationships observed can be explained using a formal description of the mod and resc functions, implying both qualitative and quantitative variations. [ABSTRACT FROM AUTHOR]

Drosophila suzukii is a pest native to Southeast Asia that causes significant economic losses to soft fruit crops. Phytosanitary irradiation is a promising treatment for D. suzukii hosts; yet an internationally recognized irradiation protocol is lacking. To fulfil specific requirements for proposing an irradiation treatment for D. suzukii, naturally infested blueberries and cherries containing a total of 37,489 late pupae were irradiated with a maximum absorbed dose of 80 Gy. Infested hosts containing a total of 9578 late pupae were considered unirradiated controls. Prevention of egg laying by females that emerged from treated pupae was considered the treatment endpoint. The fecundity and egg viability of females that emerged from treated pupae mated with their siblings were evaluated using blueberries. While females from unirradiated pupae laid a total of 43,142 eggs, no egg was laid by females that emerged from irradiated pupae. In addition, 1-day-old adults were irradiated with nominal doses of 20 and 72 Gy to evaluate whether egg laying could be prevented in flies emerging before the irradiation treatment. Females irradiated with 72 Gy laid eggs that did not hatch. Our findings suggest the minimum absorbed dose of 80 Gy as a phytosanitary irradiation treatment against D. suzukii and may support its inclusion as a treatment option in the annex of the International Standard for Phytosanitary Measures 28 (ISPM 28). [ABSTRACT FROM AUTHOR]

The introduction of non‐native species across the world represents a major global challenge. Retracing invasion origin is an important first step in understanding the invasion process, often requiring detailed sampling within the native range. Insect species frequently host Wolbachia, a widespread endosymbiotic bacterium that manipulates host reproduction to increase infected female fitness. Here, we draw on the spatial variation in infection frequencies of an actively spreading Wolbachia strain wCer2 to investigate the invasion origin of the European cherry fruit fly, Rhagoletis cerasi. This pest of cherries was introduced from Europe to North America within the last decade. First, we screen the introduced fly population for the presence of Wolbachia. The introduced populations lack the wCer2 strain and the strongly associated mitochondrial haplotype, suggesting strain absence due to founder effects with invading individuals originating from wCer2‐uninfected native population(s). To narrow down geographic regions of invasion origin, we perform spatial interpolation of the wCer2 infection frequency across the native range and predict the infection frequency in unsampled regions. For this, we use an extensive dataset of R. cerasi infection covering 238 populations across Europe over 25 years, complemented with 14 additional populations analyzed for this study. We find that R. cerasi was unlikely introduced from wCer2‐infected populations in Central and Western Europe. We propose wCer2‐uninfected populations from Eastern Europe and the Mediterranean region as the most likely candidates for the invasion origin. This work utilizes Wolbachia as an indirect instrument to provide insights into the invasion source of R. cerasi in North America, revealing yet another application for this multifaceted heritable endosymbiont. Given the prevalence of biological invasions, rapidly uncovering invasion origins gives fundamental insights into how invasive species adapt to new environments. [ABSTRACT FROM AUTHOR]

Adaptation of wild insects to an artificial laboratory environment goes hand in hand with changes in their genetic diversity and symbiotic communities. This can have a profound effect on mass rearing of insects since their biological quality and mating competitiveness might be reduced. Patterns of genetic and symbiotic variation associated with laboratory adaptation vary among species and identifying the agents of selection that drive such patterns can be extremely helpful for pest control programmes. In the present study, we profiled the genetic and symbiotic structure of a wild Drosophila suzukii population and monitored their changes during laboratory adaptation. We employed sixteen microsatellite markers and tracked the fluctuation of their frequencies as domestication was progressing. We also used 16S rRNA sequencing to depict changes in the microbiota structure. Changes in effective and observed allele numbers and heterozygosity became evident from F10, while between F0 and F6 a gradual decline was observed. The microbiota profile of D. suzukii also formed distinct clusters during adaptation as F0 and F1 were differentiated by the rest generations and presented the highest bacterial diversity. Overall, our results showed that laboratory adaptation strongly affects both genetic diversity and bacterial communities of D. suzukii. These results can serve as a reference for the design of an area-wide integrated pest management approach with a Sterile Insect Technique (SIT) component. Rearing productivity, biological quality, and mating competitiveness of a SIT mass-reared strain should be assessed in connection with genetic and symbiotic changes occurring during laboratory adaptation. [ABSTRACT FROM AUTHOR]

The European spruce bark beetle Ips typographus is a pest causing severe damage in forests dominated by Norway spruce in Europe. Microorganisms play essential roles in bark beetles, including nutrition, fitness as well as in overcoming host defenses. Here, we performed high-throughput 16S rRNA metabarcoding of I. typographus across different populations in Europe, to assess its bacterial community. We investigated four postglacial refugial areas in Europe and focused specifically on a current bark beetle hot spot in the Dolomites where we compared populations with different epidemiological phases (outbreaking vs. non-outbreaking) and across different seasons (pre-overwintering vs. overwintering). Our results show that the bacterial community structure varied among populations from the refugial areas and between different regions within the Dolomites. We found a significant difference in the bacterial community between pre-overwintering and overwintering individuals, but we did not find differences between epidemic and endemic populations. The prevalence of the genera Erwinia (which was present in all individuals) and Pseudoxanthomonas (present in almost all individuals) across all populations suggests that these taxa form the core bacterial community of I. typographus. Furthermore, several additional bacterial taxa occurred in all populations, but with variable frequencies. This study highlights a complex interaction of I. typographus and various bacterial taxa across different regions and ecological phases and provides new insights into the role of microorganisms in the biology of this important pest species. [ABSTRACT FROM AUTHOR]

Ips typographus (L.) and Pityogenes chalcographus (L.) (Coleoptera: Curculionidae) are two common bark beetle species on Norway spruce in Eurasia. Multiple biotic and abiotic factors affect the life cycles of these two beetles, shaping their ecology and evolution. In this article, we provide a comprehensive and comparative summary of selected life-history traits. We highlight similarities and differences in biotic factors, like host range, interspecific competition, host colonization, reproductive behaviour and fungal symbioses. Moreover, we focus on the species' responses to abiotic factors and compare their temperature-dependent development and flight behaviour, cold adaptations and diapause strategies. Differences in biotic and abiotic traits might be the result of recent, species-specific evolutionary histories, particularly during the Pleistocene, with differences in glacial survival and postglacial recolonization. Finally, we discuss future research directions to understand ecological and evolutionary pathways of the two bark beetle species, for both basic research and applied forest management. [ABSTRACT FROM AUTHOR]

Simple Summary: The mealworm, i.e., larvae of Tenebrio molitor, has recently been used as a substitute for conventional meat, as it has several advantages, e.g., a lower environmental impact. Knowledge of the effects of temperature and photoperiod on mealworm development is crucial to increase farming efficiency, and to contribute to a sustainable production process. As such data is scarce, we tested the effects of three different temperatures, in combination with three photoperiods, on mealworm survival, development, and growth. We found that temperature strongly affects the survival rate, developmental time, and growth rate of T. molitor larvae. Furthermore, photoperiod influences the developmental time and growth rate. The highest survival rates and growth rates, and shortest developmental times, were observed at 25 and 30 °C at constant darkness. These results are important to improve the mass rearing of mealworms for a sustainable production of food and feed. Insects are a potential substitute for conventional meat and can be part of a sustainable human diet due to their valuable nutrients and relatively low environmental production impact. One species that is already produced for human consumption and livestock feed is the mealworm, i.e., larvae of Tenebrio molitor. Knowledge of the effects of temperature, and particularly photoperiod, on mealworm development is scarce, but crucial for the improvement of rearing. Therefore, the effects of three temperatures (20 °C, 25 °C, and 30 °C), in combination with three photoperiods (long-day—16 h:8 h light:dark; short-day—8 h:16 h light:dark, and constant darkness) on mealworm survival, developmental time, and growth rate were tested. We describe a significant effect of temperature on survival rate, developmental time, and growth rate. Furthermore, significant effects of photoperiod on developmental time and growth rate were found. At 25 and 30 °C and constant darkness, the highest survival and growth rate, along with the shortest developmental time, were observed. Our data can be used to improve the mass rearing of mealworms for an efficient production of food and feed. [ABSTRACT FROM AUTHOR]

The bark beetle Ips typographus is the most destructive insect pest in Norway spruce-dominated forests. Its potential to establish multiple generations per year (multivoltinism) is one major trait that makes this beetle a severe pest. Ips typographus enters diapause to adjust its life cycle to seasonally changing environments. Diapause is characterized by developmental and reproductive arrest; it prolongs generation time and thus affects voltinism. In I. typographus a facultative, photoperiod-regulated diapause in the adult stage has been described. In addition, the presence of an obligate, photoperiod-independent, diapause has been hypothesized. The diapause phenotype has important implications for I. typographus voltinism, as populations with obligate diapausing individuals would be univoltine. To test for the presence of different I. typographus diapause phenotypes, we exposed Central and Northern European individuals to a set of photoperiodic treatments. We used two ovarian traits (egg number and vitellarium size) that are associated with gonad development, to infer reproductive arrest and thus diapause. We found a distinct effect of photoperiod on ovarian development, with variable responses in Central and Northern European beetles. We observed obligate diapausing (independent of photoperiod) individuals in Northern Europe, and both facultative (photoperiod-regulated) as well as obligate diapausing individuals in Central Europe. Our results show within-species variation for diapause induction, an adaptation to match life cycles with seasonally fluctuating environmental conditions. As the diapause phenotype affects the potential number of generations per season, our data are the basis for assessing the risk of outbreaks of this destructive bark beetle. [ABSTRACT FROM AUTHOR]