Machine learning applied to lentic habitat use by spawning walleye demonstrates the benefits of considering multiple spatial scales in aquatic research

Scale and hierarchy have received less attention in aquatic systems compared to terrestrial. Walleye Sander vitreus spawning habitat offers an opportunity to investigate scale’s importance. We estimated lake-, transect-, and quadrat-scale influences on nearshore walleye egg deposition in 28 Minnesota lakes from 2016-2018. Random forest models (RFM) estimated importance of predictive variables to walleye egg deposition. Predictive accuracies of a multi-scale classification tree (CT) and a quadrat-scale CT were compared. RFM results suggested that five of our variables were unimportant when predicting egg deposition. The multi-scale CT was more accurate than the quadrat-scale CT when predicting egg deposition. Both model results suggest that in-lake egg deposition by walleye is regulated by hierarchical abiotic processes and that silt/clay abundance at the transect-scale (reef-scale) is more important than abundance at the quadrat-scale (within-reef). Our results show machine learning can be used for scale-optimization and potentially to determine cross-scale interactions. Further incorporation of scale and hierarchy into studies of aquatic systems will increase our understanding of species-habitat relationships, especially in lentic systems where multi-scale approaches are rarely used.

Aquatain® Causes Anti-Oviposition, Egg Retention, Oocyte Melanization and Triggers Female Death in Ae. Aegypti

Background: In arboviral disease systems where the virus can be transmitted from male to female and from one generation to the next, targeting the female (especially when she is pregnant) can help alter the persistence of the virus in nature and its transmission. This is typical of Ae. aegypti, which has been unmanageable due to the development of insecticide resistance. Despite evidence that monomolecular surface films prevent the selection of genetic resistance, their potential in Aedes vector control remains largely unexplored. Methods: We examined the oviposition, egg retention, oocyte melanization, and female mortality of the Cayman Islands strain of Ae. aegypti, using choice and no-choice bioassays involving Aquatain® Mosquito Formulation (AMF). Results: When presented with similar opportunities to oviposit in two sites treated with AMF and two others with water (control), egg deposition rates were significantly higher in water than in oil presence. We also observed a matching pattern of egg deposition preference in arenas with more options in AMF-treated sites. Females laid appreciably more eggs when water was the only available medium than when all sites were treated with AMF. Also, considerably more mature eggs were withheld in the AMF no-choice arena than in the no-choice test involving only water. Internal oocyte melanization was not observed in females from the oviposition arenas with the lowest oil presence (equal-choice and water-based no-choice); in contrast, this physiological response intensified as the number of AMF-treated sites increased. Female death occurred at high rates in AMF-treated environments, and this response increased with the increasing presence of such egg deposition sites. Conclusions: This study demonstrated that AMF acted as a deterrent signal to ovipositing Ae. aegypti and an indirect adulticide. Referring to its previously reported direct toxicity on the aquatic stages of this mosquito and its biodegradability, AMF should be incorporated as a critical component in integrated control strategies for dengue and related diseases.

Responses to larval herbivory in the phenylpropanoid pathway of Ulmus minor are boosted by prior insect egg deposition

Main conclusion Elms, which have received insect eggs as a ‘warning’ of larval herbivory, enhance their anti-herbivore defences by accumulating salicylic acid and amplifying phenylpropanoid-related transcriptional and metabolic responses to hatching larvae. Abstract Plant responses to insect eggs can result in intensified defences against hatching larvae. In annual plants, this egg-mediated effect is known to be associated with changes in leaf phenylpropanoid levels. However, little is known about how trees—long-living, perennial plants—improve their egg-mediated, anti-herbivore defences. The role of phytohormones and the phenylpropanoid pathway in egg-primed anti-herbivore defences of a tree species has until now been left unexplored. Using targeted and untargeted metabolome analyses we studied how the phenylpropanoid pathway of Ulmus minor responds to egg-laying by the elm leaf beetle and subsequent larval feeding. We found that when compared to untreated leaves, kaempferol and quercetin concentrations increased in feeding-damaged leaves with prior egg deposition, but not in feeding-damaged leaves without eggs. PCR analyses revealed that prior insect egg deposition intensified feeding-induced expression of phenylalanine ammonia lyase (PAL), encoding the gateway enzyme of the phenylpropanoid pathway. Salicylic acid (SA) concentrations were higher in egg-treated, feeding-damaged leaves than in egg-free, feeding-damaged leaves, but SA levels did not increase in response to egg deposition alone—in contrast to observations made of Arabidopsis thaliana. Our results indicate that prior egg deposition induces a SA-mediated response in elms to feeding damage. Furthermore, egg deposition boosts phenylpropanoid biosynthesis in subsequently feeding-damaged leaves by enhanced PAL expression, which results in the accumulation of phenylpropanoid derivatives. As such, the elm tree shows similar, yet distinct, responses to insect eggs and larval feeding as the annual model plant A. thaliana.

Host tissue proteomics reveal insights into the molecular basis of Schistosoma haematobium-induced bladder pathology

Background: Urogenital schistosomiasis remains a major public health concern worldwide. In response to egg deposition, the host bladder undergoes gross and molecular morphological changes relevant for disease manifestation. However, limited mechanistic studies to date imply that the molecular mechanisms underlying pathology have not been well-defined. We leveraged a mouse model of urogenital schistosomiasis to perform for the first time, proteome profiling of the early molecular events that occur in the bladder after exposure to S. haematobium eggs, and to elucidate the protein pathways involved in urogenital schistosomiasis-induced pathology. Methods: Purified S. haematobium eggs or control vehicle were microinjected into the bladder walls of mice. Mice were sacrificed seven days post-injection and bladder proteins isolated and processed for proteome profiling using mass spectrometry. Results: We demonstrate that biological processes including carcinogenesis, immune and inflammatory responses, increased protein translation or turnover, oxidative stress responses, reduced cell adhesion and epithelial barrier integrity, and increased glucose metabolism were significantly enriched in S. haematobium infection. Conclusion: S. haematobium egg deposition in the bladder results in significant changes in proteins and pathways that play a role in pathology. Our findings highlight the potential bladder protein indicators for host-parasite interplay and provide new insights into the complex dynamics of pathology and characteristic bladder tissue changes in urogenital schistosomiasis. The findings will be relevant for development of improved interventions for disease control.

Olfactory Responses of Trissolcus Mitsukurii to Plants Attacked by Target and Non-target Stink Bugs Suggest Low Risk for Biological Control

In crop systems, successful management of invasive insect herbivores can be achieved through the introduction of exotic biocontrol agents, i.e. parasitoids or predators, having a coevolutionary history with the pest. To avert threats to local biodiversity, recent legislations require a risk assessment for the organism to be released. Evaluation of its ability to exploit, for host location, odours associated with target and non-target species is crucial for a better definition of its ecological host range. Using Y-tube olfactometer in quarantine laboratory, we investigated the ability of the Asiatic egg parasitoid Trissolcus mitsukurii (Hymenoptera: Scelionidae) to exploit odours associated with the global invader Halyomorpha halys (Hemiptera: Pentatomidae) and with non-target stink bugs native to Southern Europe. We demonstrated that T. mitsukurii is attracted by plants exposed to feeding and egg deposition of the coevolved H. halys or the native Nezara viridula, while it is not attracted by physogastric females or eggs alone. Remarkably, T. mitsukurii is repelled by plants bearing eggs of the beneficial stink bug, Arma custos. Our results usefully contribute to a more thorough and nuanced assessment of the potential non-target risks in case of massive parasitoid release.

A rise-to-threshold signal for a relative value deliberation

Whereas progress has been made in identifying neural signals related to rapid, cued decisions, less is known about how brains guide and terminate more ethologically relevant deliberations, where an animal's own behavior governs the options experienced over minutes. Drosophila search for many seconds to minutes for egg-laying sites with high relative value and neurons, called oviDNs, exist whose activity fulfills necessity and sufficiency criteria for initiating the egg-deposition motor program. Here we show that oviDNs express a calcium signal that rises over seconds to minutes as a fly deliberates whether to lay an egg. The calcium signal dips when an egg is internally prepared (ovulated), rises at a rate related to the relative value of the current substrate being experienced, and reaches a consistent peak just prior to the abdomen bend for egg deposition. We provide perturbational evidence that the egg-deposition motor program is initiated once this signal hits a threshold and that sub-threshold variation in the signal regulates the time spent deliberating and, ultimately, the option chosen. These results argue that a rise-to-threshold signal guides Drosophila to lay eggs on substrate options with high relative value, with each egg-laying event representing a self-paced decision similar to real-world decisions made by humans and other mammals.

Neuronal control of suppression, initiation and completion of egg deposition in Drosophila melanogaster

Egg-laying in Drosophila is the product of post-mating physiological and behavioural changes that culminate in a stereotyped sequence of actions. While egg-laying behaviour has been mostly used as a system to understand the neuronal basis of decision making in the context of site selection, it harbours a great potential as a paradigm to uncover how, once a site is selected, the appropriate motor circuits are organized and activated to deposit an egg. To study this programme, we first describe the different stages of the egg-laying programme and the specific actions associated with each stage. Using a combination of neuronal activation and silencing experiments we characterize the role of three distinct neuronal populations in the abdominal ganglion with different contributions to the egg deposition motor elements. Specifically, we identify a subset of glutamatergic neurons and a subset of cholinergic neurons that promote the initiation and completion of egg expulsion respectively, while a subset of GABAergic neurons suppresses egg-laying. This study provides insight into the organization of neuronal circuits underlying complex motor behaviour.

Pine defense responses to eggs of an herbivorous sawfly are elicited by an annexin-like protein

Known elicitors of plant defenses against eggs of herbivorous insects are low-molecular-weight organic compounds associated with the eggs. However, previous studies provided evidence that also proteinaceous compounds present in secretion associated with eggs of the herbivorous sawfly Diprion pini can elicit defensive responses in Pinus sylvestris. Pine responses induced by the proteinaceous secretion are known to result in enhanced emission of (E)-β-farnesene, which attracts egg parasitoids killing the eggs. Here, we aimed to identify the defense-eliciting protein and elucidate its function. After isolating the defense-eliciting protein from D. pini egg secretion by ultrafiltration and gel electrophoresis, we identified it by MALDI-ToF mass spectrometry as an annexin-like protein, which we named “diprionin”. Further GC-MS analyses showed that pine needles treated with heterologously expressed diprionin released enhanced quantities of (E)-β-farnesene. Our bioassays confirmed attractiveness of diprionin-treated pine to egg parasitoids. Expression of several pine candidate genes involved in terpene biosynthesis and regulation of ROS homeostasis was similarly affected by diprionin and natural sawfly egg deposition. However, the two treatments had different effects on expression of pathogenesis related genes (PR1, PR5). Diprionin is the first egg-associated proteinaceous elicitor of indirect plant defense against insect eggs described so far.

Olfactory response of Trichogramma pretiosum (Hymenoptera: Trichogrammatidae) to volatiles induced by transgenic maize

Plants not only respond to herbivorous damage but adjust their defense system after egg deposition by pest insects. Thereby, parasitoids use oviposition-induced plant volatiles to locate their hosts. We investigated the olfactory behavioral responses of Trichogramma pretiosum Riley, 1879 (Hymenoptera: Trichogrammatidae) to volatile blends emitted by maize (Zea mays L.) with singular and stacked events after oviposition by Spodoptera frugiperda Smith, 1797 (Hymenoptera: Trichogrammatidae) moths. Additionally, we examined possible variations in gene expression and on oviposition-induced volatiles. We used a Y-tube olfactometer to test for the wasp responses to volatiles released by maize plants oviposited by S. frugiperda and not-oviposited plants. Using the real-time PCR technique (qRT-PCR), we analyzed the expression of lipoxygenase and three terpene synthases genes, which are enzymes involved in the synthesis of volatile compounds that attract parasitoids of S. frugiperda. Olfactometer tests showed that T. pretiosum is strongly attracted by volatiles from transgenic maize emitted by S. frugiperda oviposition (VTPRO 3, more than 75% individuals were attracted). The relative expression of genes TPS10, LOX e STC was higher in transgenic hybrids than in the conventional (isogenic line) hybrids. The GC-MS analysis revealed that some volatile compounds are released exclusively by transgenic maize. This study provides evidence that transgenic hybrids enhanced chemical cues under oviposition-induction and helped to increase T. pretiosum efficiency in S. frugiperda control. This finding shows that among the evaluated hybrids, genetically modified hybrids can improve the biological control programs, since they potentialize the egg parasitoid foraging, integrating pest management.

Traumatic mating increases anchorage of mating male and reduces female remating duration and fecundity in a scorpionfly species

Traumatic mating is the male wounding his mate during mating using specialized anatomy. However, why males have evolved to injure their mates during mating remains poorly understood. We studied traumatic mating in Dicerapanorpa magna to determine its effects on male and female fitness. The sharp teeth on male gonostyli penetrate the female genitalia and cause copulatory wounds, and the number of scars on the female genitals is positively related to the number of times females mated. When the injurious teeth were encased with low-temperature wax, preventing their penetration of the female's genitalia during mating, male mating success and copulation duration were reduced significantly, indicating the importance of the teeth in allowing the male to secure copulation, remain in copula and effectively inseminate his mate. The remating experiments showed that traumatic mating had little effect on the female mating refractory period, but significantly reduced female remating duration with subsequent males, probably benefiting the first-mating male with longer copulation duration and transferring more sperm into the female's spermatheca. The copulatory wounds reduced female fecundity, but did not accelerate the timing of egg deposition. This is probably the first report that traumatic mating reduces female remating duration through successive remating experiments in animals. Overall, our results provide evidence that traumatic mating in the scorpionfly helps increase the male's anchoring control during mating and provides him advantage in sperm competition, but at the expense of lowering female fecundity.