Characterisation of the Alarm Pheromone of Bathycoelia Distincta (Pentatomidae).

Bathycoelia distincta (Pentatomidae) is the dominant pest in South African macadamia orchards, where adults are responsible for causing severe yield losses. Similar to other hemipterans, B. distincta release volatile compounds from scent glands that can deter natural enemies and act as an alarm signal among conspecifics. The overall aim of this study was to characterise the alarm pheromone of B. distincta. We: (i) analysed the scent gland contents of individual adult B. distincta by gas chromatography-mass spectrometry (GC-MS), (ii) quantified volatiles released from live stink bugs after stress, and (iii) evaluated the electrophysiological and behavioural activity of alarm pheromone compounds with dose-response experiments. A blend of fourteen compounds was identified in the scent gland extracts of adult stink bugs. Of these, six compounds were detected in the effluvia of live stressed stink bugs [(E)-2-hexenal, (E)-2-decenal, tridecane, dodecane, (E)-4-oxohex-2-enal and (E)-2-decenyl acetate]. No qualitative or quantitative differences were observed between sexes. Tridecane was the most abundant compound, comprising ∼50% of total secretions. Only (E)-2-hexenal, (E)-2-decenal, and (E)-4-oxohex-2-enal elicited an antennal response in both sexes. Finally, exposure to a mixture of (E)-2-hexenal, (E)-2-decenal, and (E)-4-oxohex-2-enal resulted in an increase in the speed and distance travelled by walking bugs and a decrease in time spent resting compared to unexposed bugs. Our results show that the blend of (E)-2-hexenal, (E)-2-decenal, and (E)-4-oxohex-2-enal can induce an alarm response in B. distincta.

Sirex woodwasp: a model for evolving management paradigms of invasive forest pests.

The Sirex woodwasp, Sirex noctilio, and its fungal mutualist, Amylostereum areolatum, together constitute one of the most damaging invasive pests of pine. Despite a century of research and well-established management programs, control remains unpredictable and spread continues to new areas. Variable success in managing this pest has been influenced by complex invasion patterns, the multilayered nature of biological interactions, the varying local ecologies, and microevolutionary population processes in both the biocontrol organisms and in the wasps. Recent research findings are challenging the historical perspectives on methods to manage the Sirex woodwasp, calling for management programs to incorporate the variable local dynamics affecting this pest complex. In this regard, the Sirex woodwasp provides a superb model to illustrate the need for a different approach to develop efficient and sustainable management tools to deal with the growing and global nature of pest invasions in forests and plantations.

Lack of fidelity revealed in an insect-fungal mutualism after invasion.

Symbiont fidelity is an important mechanism in the evolution and stability of mutualisms. Strict fidelity has been assumed for the obligate mutualism between Sirex woodwasps and their mutualistic Amylostereum fungi. This assumption has been challenged in North America where the European woodwasp, Sirex noctilio, and its fungal mutualist, Amylostereum areolatum, have recently been introduced. We investigate the specificity of the mutualism between Sirex and Amylostereum species in Canada, where S. noctilio co-infests Pinus with native Sirex nigricornis and its mutualist, Amylostereum chailletii. Using phylogenetic and culture methods, we show that extensive, reciprocal exchange of fungal species and strains is occurring, with 75.3 per cent of S. nigricornis carrying A. areolatum and 3.5 per cent of S. noctilio carrying A. chailletii. These findings show that the apparent specificity of the mutualism between Sirex spp. and their associated Amylostereum spp. is not the result of specific biological mechanisms that maintain symbiont fidelity. Rather, partner switching may be common when shifting geographical distributions driven by ecological or anthropogenic forces bring host and mutualist pairs into sympatry. Such novel associations have potentially profound consequences for fitness and virulence. Symbiont sharing, if it occurs commonly, may represent an important but overlooked mechanism of community change linked to biological invasions.

Biology and rearing of Cleruchoides noackae (Hymenoptera: Mymaridae), an egg parasitoid for the biological control of Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae).

Cleruchoides noackae Lin and Huber (Hymenoptera: Mymaridae) is a solitary egg parasitoid of Thaumastocoris peregrinus Carpintero and Dellapé (Hemiptera: Thaumastocoridae). The parasitoid was first described in 2009 and its biology and rearing are poorly understood. A key obstacle to the use of C. noackae as a biological control agent has been the ability to consistently rear it under quarantine conditions. This study reports on a series of experiments conducted in quarantine to rear C. noackae and to examine the effects of diet on longevity, per capita reproduction, and progeny sex ratio, as well as to determine development time, and preference and suitability of host eggs of different ages. When supplemented with honey solution, the longevity of C. noackae females increased significantly by 2.4 d and that of males by 1.7 d, relative to the unfed adults. Mean per capita reproduction for the honey-fed wasps was 7.7 offspring per female, with progeny sex ratio slightly skewed toward males. Mean percentage parasitism was 32.2%. C. noackae was capable of parasitizing and completing development from oviposition to adult eclosion within 15.7 d in host eggs between 0 and 5 d old. The ability of C. noackae to parasitize a wide range of host egg ages increases the period of vulnerability of T. peregrinus to attack, increasing its potential efficacy as a biological control agent. The methods and results reported here represent a crucial step in the ongoing efforts to develop this potential biological control system.

Efficacy of Commercially Available Entomopathogenic Agents against the Polyphagous Shot Hole Borer in South Africa.

The invasive ambrosia beetle, Euwallacea fornicatus, was first reported in South Africa in 2018. The beetle has now spread to eight provinces of the country and has had a devastating impact on both native and non-native tree species. This is especially true for trees located in urban and peri-urban environments. Recent predictions are that the South African E. fornicatus invasion will cost an estimated ZAR 275 billion (approx. USD 16 billion) if it continues to spread uncontrollably, justifying an urgent need for its effective management in the country. One option is biological control, which is preferred over the use of chemicals due to its lower environmental impact. We tested two broad-spectrum fungal entomopathogenic agents, Eco-Bb® and Bio-Insek, which are commercially available in South Africa, for efficacy against E. fornicatus. Initial laboratory assays yielded promising results. However, beetle infestation trials using treated pieces of woody castor bean stems showed little effect on beetle survival and reproduction.

Genetic diversity of the two-spotted stink bug Bathycoelia distincta (Pentatomidae) associated with macadamia orchards in South Africa.

The South African macadamia industry is severely affected by a complex of stink bugs, dominated by the two-spotted stink bug, Bathycoelia distincta Distant (Pentatomidae). This species was first discovered during the spring of 1984 in the Limpopo province. Although considerable effort has been spent trying to manage this pest, it continues to be a pest of concern for the macadamia industry. Information on the genetic diversity of this species is lacking, despite the potential relevance of such information for management strategies. The present study aimed to characterise the genetic diversity of B. distincta populations in South Africa. The Cytochrome c Oxidase Subunit 1 (COI) and cytochrome b (Cytb) gene regions were sequenced from individuals collected from the three main regions of macadamia production over three different seasons (2018-2020). An overall high haplotype diversity (COI = 0.744, Cytb = 0.549 and COI+Cytb = 0.875) was observed. Pairwise mean genetic distance between populations from each region varied from 0.2-0.4% in both datasets, which suggests the absence of cryptic species. The median joining network for both datasets consisted of one or two central haplotypes shared between the regions in addition to unique haplotypes observed in each region. Finally, low genetic differentiation (FST < 0.1), high gene flow (Nm > 1) and the absence of a correlation between genetic and geographic distance were estimated among populations. Overall, these results suggest that the B. distincta populations are not structured among the areas of macadamia production in South Africa. This might be due to its ability to feed and reproduce on various plants and its high dispersal (airborne) between the different growing regions of the country along with the rapid expansion of macadamia plantations in South Africa.

Diversity and Molecular Barcoding of Stink Bugs (Hemiptera: Pentatomidae) Associated with Macadamia in South Africa.

Stink bugs are major pests of macadamia in South Africa. Accurate identification and knowledge of species composition are important to inform management practices. The overall aims of this study were to identify stink bug species from macadamia orchards in South Africa using morphology, and to establish a DNA database based on the cytochrome c oxidase subunit 1 gene region. A total of 21 stink bug species were found in macadamia orchards in KwaZulu-Natal, Limpopo and Mpumalanga provinces. Bathycoelia distincta Distant, 1878, was the dominant species throughout all three growing regions. Two unidentified species of Boerias Kirkaldy, 1909, here designated as Boerias sp. 1 and Boerias sp. 2, were the second and third most abundant species found in KwaZulu-Natal. No species of Boerias has previously been reported in association with macadamia. Evidence of a cryptic third species of Boerias was also found. Species composition fluctuated over three growing seasons in Limpopo and differed between the three growing regions during the 2019-2020 season, highlighting the need for ongoing monitoring of these important pest species. The DNA barcode database developed in this study will be valuable for future monitoring and identifications, including cryptic or polymorphic stink bug species and different life stages.

Biotic and abiotic effects on density, body size, sex ratio, and survival in immature stages of the European woodwasp, Sirex noctilio.

Resource quality can have direct or indirect effects on female oviposition choice, offspring growth and survival, and ultimately on body size and sex ratio. We examined these patterns in Sirex noctilio Fabricus, the globally invasive European pine woodwasp, in South African Pinus patula plantations. We studied how tree position as well as natural variation in biotic and abiotic factors influenced sex-specific density, larval size, tunnel length, male proportion, and survival across development. Twenty infested trees divided into top, middle, and bottom sections were sampled at three time points during larval development. We measured moisture content, bluestain fungal colonization, and co-occurring insect density and counted, measured, and sexed all immature wasps. A subset of larval tunnels was measured to assess tunnel length and resource use efficiency (tunnel length as a function of immature wasp size). Wasp density increased from the bottoms to the tops of trees for both males and females. However, the largest individuals and the longest tunnels were found in bottom sections. Male bias was strong (~10:1) and likewise differed among sections, with the highest proportion in the middle and top sections. Sex ratios became more strongly male biased due to high female mortality, especially in top and middle sections. Biotic and abiotic factors such as colonization by Diplodia sapinea, weevil (Pissodes sp.) density, and wood moisture explained modest residual variation in our primary mixed effects models (0%-22%). These findings contribute to a more comprehensive understanding of sex-specific resource quality for S. noctilio and of how variation in key biotic and abiotic factors can influence body size, sex ratio, and survival in this economically important woodwasp.

Mechanisms that influence sex ratio variation in the invasive hymenopteran Sirex noctilio in South Africa.

Sirex noctilio is an economically important invasive pest of commercial pine forestry in the Southern Hemisphere. Newly established invasive populations of this woodwasp are characterized by highly male-biased sex ratios that subsequently revert to those seen in the native range. This trend was not observed in the population of S. noctilio from the summer rainfall regions in South Africa, which remained highly male-biased for almost a decade. The aim of this study was to determine the cause of this persistent male bias. As an explanation for this pattern, we test hypotheses related to mating success, female investment in male versus female offspring, and genetic diversity affecting diploid male production due to complementary sex determination. We found that 61% of females in a newly established S. noctilio population were mated. Microsatellite data analysis showed that populations of S. noctilio from the summer rainfall regions in South Africa are far less genetically diverse than those from the winter rainfall region, with mean Nei's unbiased gene diversity indexes of 0.056 and 0.273, respectively. These data also identified diploid males at low frequencies in both the winter (5%) and summer (2%) rainfall regions. The results suggest the presence of a complementary sex determination mechanism in S. noctilio, but imply that reduced genetic diversity is not the main driver of the male bias observed in the summer rainfall region. Among all the factors considered, selective investment in sons appears to have the most significant influence on male bias in S. noctilio populations. Why this investment remains different in frontier or early invasive populations is not clear but could be influenced by females laying unfertilized eggs to avoid diploid male production in populations with a high genetic relatedness.