MULTIDISCIPLINARY APPROACH TO BIOACOUSTICS: Integrating phylogenomics, biophysics, and functional genomics to unravel the evolution of hearing and sin

Lead Research Organisation: University of Lincoln
Department Name: School of Life Sciences

Abstract

Acoustic communication is widespread across the animal kingdom. For decades, studies of acoustically communicating animals have collectively revealed the intricate interplay and co-evolution between signal sender and receiver, as well as the diversity of morphological structures involved in hearing and sound production and their complex neurophysiological underpinnings. However, the current approach in the field of bioacoustics is often narrowly focused on a particular aspect of acoustic communication using single model organisms, making it challenging to draw a general conclusion about how singing and hearing have been shaped through time. This project will take an explicitly clade-based approach using a comprehensive dated phylogeny of Ensifera, the most diverse group of acoustically communicating animals, complemented with cutting-edge techniques in imaging, biophysics, and functional genomics to evaluate mechanisms of hearing and singing in a comparative framework. The ambitious scope of the proposed phylogenomic analysis will firmly establish evolutionary relationships among and within the major ensiferan lineages, which have remained elusive for many decades. The project will generate an unprecedented amount of detailed morphological, biophysical, and genetic data using X-ray micro and nano computed tomography, microscanning laser Doppler vibrometry, RNA-seq, and target capture sequencing. The results from this project will answer outstanding questions about when acoustic communication originated and how often it has been lost, how lineages independently evolved different mechanisms of hearing and signalling, what the physical, neural and genetic bases of these are, and how signal diversity is causally related to diversification and speciation. These findings will collectively reveal general insights about the evolution of acoustic communication.

Planned Impact

This project will fill important gaps in biodiversity and bioacoustics knowledge and provide unique educational opportunities at multiple academic levels, from K-12 students to postdoctoral researchers. It will place particular emphasis on broadening participation of underrepresented students, all of whom stand to collectively benefit from our exciting scientific discoveries. The project results will be broadly disseminated through open access peer-reviewed journals, conference presentations, as well as popular media outlets. The international project team will establish a strong collaborative network that will persist long after the grant period. Broader impacts activities include: (1) Develop and offer two rounds of 10-week Summer REU Program specifically targeting Polynesian and Hispanic undergraduate students to provide hands-on experience in biodiversity and genomics research; (2) Develop an outreach activity about insect acoustic communication targeting K-12 students called "The Night at The Gardens" at TAMU educational gardens; (3) Develop an interactive exhibit about the evolution and the genetic basis of acoustic communication to be showcased at the Royal Society Summer Science Exhibition in the U.K.; and (4) Develop a "Cricket Course" targeting the general public and scientists to provide training in identification, ecology, behavior, and bioacoustics of katydids, crickets, and allies at the Archbold Biological Station in Florida and the Soltis Center for Research and Education in Costa Rica.

Benefits to the UK population are also envisaged via potential developments to sensors based on the acoustic organs of the. This exploitation will be targeted at UK industry through the research groups of the principal applicant (PA) at Lincoln. This links with the Research & Enterprise department (RID) and the Laser Group at Lincoln.
 
Description Progress in the sound generator (the wings)
The article titled 'Reviving the sound of a 150-year-old insect: the bioacoustics of Prophalangopsis obscura (Ensifera: Hagloidea)' is currently under review at PLOS One. This article uses a multi-disciplinary approach to reconstructing the acoustic signals of a dead museum specimen, with a unique phylogenetic position within the Ensifera. Reviving sound in this species will allow evolutionary comparisons to extinct ensiferans from the Jurassic, and advance models for acoustic reconstruction across the Ensifera.

Progress in the hearing system
The use of micro-rheology to infer mechanics in the katydid inner ear fluid
Copiphora gorgonensis is one of the model species used, and have one of the most highly elaborated ears in insects, which allow them to detect a wide range of acoustic stimuli, from the calls of their mates to the echolocating shrieks of bats that predate on them. Within these ears, located in their front legs, are endowed with outer, middle and inner ear parts, just as in the human ear. Within the inner ear (the insect cochlea) there is a vesicle full of an undetermined fluid. These fluid covers the sensors that detect sound-induced vibrations in the fluid and convert them into electrical impulses and information. The way in which vibrations reach these sensors may depend on the physical properties of the fluid. We are exploring these properties, using a sophisticated technique called microrheology that allows us to characterize the deformations of very small volumes of fluid in response to tiny physical perturbations. So far, using this technique, we have seen that this fluid is much more viscous than water and has not elasticity whatsoever.

In Copiphoras the cochlear fluid is physically isolated from the circulatory system. This implies that motions and physical perturbations that occur in either of the systems do not affect the other one.
We have observed high variability in the rheological properties of the AV fluid. We are still assessing how much of this variability is due to the precision of the microrheological method, but in all measurements the viscosity was at least twice as high as in water.
At the wide range of frequencies we have explored (Ø10-2-104Hz), the fluid in the auditory vesicle behaves Newtonianly. This means that no significant elastic component was detected, which has implications for the transmission of traveling waves.

Travelling waves in the tympanic membrane of a primitive cricket
The article titled 'Auditory mechanics in the Grig (Cyphoderris monstrosa): Tympanal travelling waves and frequency discrimination as a precursor to inner ear tonotopy' is currently under review as an invited resubmission at Proceedings B. This article uses micro-CT scanning and micro-scanning laser-doppler vibrometry to investigate ear function in a relict ensiferan, and compares to the auditory mechanics of modern species to infer how the crista acustica and hearing process has changed over evolutionary time.
Exploitation Route Key findings in hearing
The use of 3D printed ear models
The 3D model method that we have developed has wide biophysical applications beyond insect science, including advancing ultrasonic detection (e.g. for cetacean conservation, military application), and further biophysical investigations across other taxa.

The use of Micro-rheology
Results of this part will be published in scientific journals. In addition, we expect that the methodology we are developing will be invaluable in other fields in which the rheology of tiny volumes of fluid needs to be ascertained (e.g. microbial ecology, physiology, biomedecine...), and thus we aim to publish the protocols as well as the Matlab code generated in online open repositories by the end of the present year.
This work is targeting Objective 2 (Evolution of ensiferan hearing organs and auditory signal processing mechanisms), and is part of the Research Approach 4.3 (Morphological and biophysical data generation), where we aim to measure the inner ear response to tympanal vibrations. Results are very encouraging and proceed at a good pace, but the work is still ongoing, as the researcher responsible for it (Òscar Guadayol) started his contract on August last year.

Key findings in sound production mechanisms
Advanced method for sound reconstruction may allow further extinct taxa sounds to be reconstructed in the future, to advance our understanding of ancient soundscapes.
Sectors Digital/Communication/Information Technologies (including Software),Education,Culture, Heritage, Museums and Collections
 
Description Part of the impact emerged from this research relates to politic and policy makers, as described in the section Influence on Policy, Practice, Patients & the Public, for the Havana syndrome and the new legislations of the US government in response to the Sonic Attack situation. But our use of 3D printed geometries is having considerable impact on the use of Museum specimens for peripheral hearing research, and we envisage a strong impact on the 3Rs, in the reduction, refinement and replacement of animals in scientific research.
First Year Of Impact 2022
Sector Environment,Government, Democracy and Justice,Culture, Heritage, Museums and Collections,Security and Diplomacy
 
Description The impact and relevance of bioacoustics research in international politics: the Havana syndrome
Geographic Reach North America 
Policy Influence Type Citation in other policy documents
Impact In late 2016, US diplomats in Cuba started experiencing unexplained health problems including tinnitus, vertigo, and cognitive difficulties. Some complained of a preceding "shrill sound" in their homes, which left experts worried that a "sonic attack" might have caused their ailments. The US State Department determined that the incidents were "specific attacks", and responded by cutting its Cuban embassy staff by 60% and expelling 15 Cuban diplomats from Washington, all despite Cuba's fervent denials of wrongdoing. This mysterious case has seen US-Cuba relations plummet from what had been a high point in 2015 when the two countries, estranged for half a century, restored diplomatic ties under President Obama. The high-pitched sound linked to these attacks was recorded by US personnel in Cuba and released by the Associated Press (AP) in 2016. Reviewing US evidence and recording their own data, Cuban scientists suggested in 2017 that the noises may have stemmed from a chirping Jamaican field cricket, a common insect in Cuba. This was dismissed by Washington as the short chirp of the cricket did not match the abrasive continuous drone recorded by US personnel. The associated ERC grant "The Insect Cochlea" involves a broad bioacoustics approach to crickets and bush-crickets in the developmental Work Packages, hence this problem brought the attention of the PI and a collaborator in the USA. Convinced that the AP sound was produced by an insect, they searched databases for acoustic recordings of Caribbean insects, and produced acoustic experiments and simulations to find that the AP recording matched the calling song of the Indies short-tailed cricket Anurogryllus celerinictus. Our findings, available in BioRxiv, caused much debate, and extensive efforts at the US National Academy of Sciences to contradict our findings. Finally, a group of elite scientists, advising the US government, supported our work, which led to President Biden signing the Havana Act in 2021.
URL https://www.documentcloud.org/documents/21068770-jason-report-2018-havana-syndrome
 
Title Cloning out sequences of mechanosensory proteins 
Description Bush crickets (katydids) are relatively large insects, which communicate over long distances by singing across a range of frequencies, including the ultrasonic range in some species. Signal receivers perceive these songs through two complex ears, each of which consists of two tympanic membranes on one of the front legs coupled to a spiracular opening on the side of the thorax via a tracheal tube. Between the tympana on each front leg lies a line of sensory cells called the crista acustica, which converts the travelling fluid waves, themselves a decelerated representation of the outside sound waves, into neural signals that are then transmitted to and interpreted by the central nervous system. Each cell of the crista acustica contains mechanosensory proteins that play a key role in converting the mechanical information of the fluid waves into a neural signal. We have had some success in cloning out sequences for a range of mechanosensory proteins that potentially play a role in bush cricket hearing. We now have partial sequences for nompC, a Piezo channel, and a Prestin-like protein, all for our transparent study species, Phlugis, which we would ordinarily collect from field sites in South America. We also aim to obtain sequence data for Nanchung and Inactive, and to also find the homologous sequences in Mecopoda and Copiphora, which are easier to rear in laboratory culture and will allow us to carry on with our project despite COVID travel restrictions. These sequences will allow a molecular-based approach to investigating the molecular functioning of the ear, including the use of RNAi, qPCR and immunohistochemistry (antibody labelling). The sequences that we already have for Phlugis will allow this approach to begin once we have sufficient numbers in culture. 
Type Of Material Biological samples 
Year Produced 2020 
Provided To Others? No  
Impact We expect that by developing the katydid hearing system as a model for acoustic communication and hearing, we will contribute towards a reduced reliance on mammalian (particularly rodent) animal use in invasive experiments. This will be in line with the increased need to reduce the number of vertebrates used in experimental systems, for which Home Office licences are needed. It is possible to investigate the katydid ear in a non-lethal way, because the most interesting features are on the front leg. This is very different from equivalent experiments in rodents, which obviously target the skull and require that the animal be euthanized immediately after data collection. 
 
Title Confocal Microscopy of Taenidia Fibers in the ear canal of katydid and crickets 
Description The method provides the protocol to generate high resolution fluorescence images of the molecular composition of single taenidia fibers using confocal laser scanning. Current focus of the study is to investigate resilin and chitin, however the protocol can be applied to any monolayer of trachea fibers with slight adjustments on filter settings. The method aims to identify formation of chitin nanofilaments in the interior of the taenidia, estimate the amount of relisin protein in relation to chitin of a single fiber, and investigate the material composition of the part of the fiber that forms the interior surface of the acoustic trachea. Our approach includes a comprehensive dissection protocol to prepare microsamples of fixed taenidia fiber monolayers and mounting on standard glass slides under specific orientation where the interior surface of the fibers can be identified. Options for autofluorescence and stainning are thoroughly derscribed. By modifying the confocal aperture and adjustments in the pinhole, thin sections of the sample (500nm) supporting the interior structure of the trachea can be imaged with increased resolution. Using z-plane stacking composite images (2D, 3D) of chitin and resilin at different molecular planes can be generated. 
Type Of Material Biological samples 
Year Produced 2022 
Provided To Others? No  
Impact The spatial relation between chitin and resilin in single taenidia fibers can be elucidated with the potential to clarify the molecular interplay between the rigidity and elasticity in correlation with nanoscale mechanical data and force measurements. The method can contribute to the understanding of evolutionary development of multifactional biological materials that could lead to significant development of novel biomaterials 
 
Title Confocal microscopy of katydid hearing organ 
Description We have used traditional paraformaldehyde fixation and histological staining combined with confocal microscopy to investigate the cellular and extracellular organisation of the katydid hearing organ. Antibodies, stains and/or autofluorescence for Y-tubulin, F-actin, chitin, resilin and extracellular membranes have been applied to fixed tissues, and imaged across 2 micron slices before 3D reconstruction. 
Type Of Material Biological samples 
Year Produced 2022 
Provided To Others? No  
Impact We have visualised katydid scolopidial (mechanosensory) cells, including the cilia, at a level of detail never seen before, which should open avenues for investigating the molecular and cellular nature of novel sensory systems. 
 
Title New histological technique in bush crickets ears 
Description We have started to optimise a recent histological technique in bush crickets. Known as DEEP-Clear (Pende et al., 2020), this process removes pigmentation from fixed tissues to allow for high-quality microscopic imaging. This will allow us to characterise the structure of the katydid ear using both histological stains and immunohistochemistry. This will be particularly useful for assessing the properties of the tectorial membrane, which is attached to the cells of the crista acustica but remains a relatively uncharacterised structure within the insect ear. We are currently optimising DEEP-Clearing in lab-reared katydids, especially the large Copiphora species. We also spent a week in Didcot at the Diamond Light Source national synchrotron facility where we performed pilot studies for future imaging projects. We aim to go back to DLS at some point in 2021, depending on the COVID-19 situation throughout the year. 
Type Of Material Biological samples 
Year Produced 2021 
Provided To Others? No  
Impact By developing the katydid hearing system as a model for acoustic communication and hearing, we can potentially reduce the number of vertebrates used in experimental systems, which require Home Office licences and specialised rearing facilities. 
 
Title Rheological characterization of ear fluids and hemolymph in Ensifera 
Description Multiple particle tracking microrheology (MPTM) of the hemolymph (1) and the fluid acoustic vesicle (2). The fluid of interest is mixed with very small volumes (~1 pL) of a highly concentrated aqueous suspension of 1.02µm fluorescent microspheres (Fluoresbrite® YG Carboxylate Microspheres, Polysciences Inc.). Samples are then placed on an inverted epifluorescence microscope (Zeiss AxioVert A1 Fl), which is equipped with an LED source (CoolLED pE-300 white) and a temperature controlling stage and mounted on an antivibration table to eliminate environmental noise. Videos are then recorded at 00X magnification with a long-distance objective (ZeissLD Plan-Neofluor 40x/0.6NA Corr) with an ultra high speed camera (Photron FASTCAM MINI UX100). Microspheres are tracked as they undergo Brownian motion using in-house Matlab code. From the frequency-dependent mean square displacements (MSD) it is possible to construct spectra of the viscoelastic moduli of the fluid of interest. Currently, using this setup we have been able to obtain spectra spanning more than 4 orders of magnitude of frequency (from 1 to 50000Hz). In the case of hemolymph (1) the sample is extracted from the specimen using a capillary previously seeded with the suspension of microspheres, and visualized directly on the capillary. To minimize optical distortions we used capillaries with a square cross-section (CM scientific). In the case of the fluid in the acoustic vesicle, the volume is too small (ØnL) to perform an extraction. To overcome this we have develop a protocol to perform microrheology in situ and in vivo. The suspension of microspheres is injected into the vesicle using a pneumatic microinjection system (PV830 Picopump, WPI). Capillaries are previously pulled using a P-97 micropipette puller (Sutter Inc.). Each capillary was calibrated individual before its use by injecting the microspheres suspension within mineral oil and measuring the diameter of the droplets. The specimen is then placed under the microscope. The cuticle in these species is transparent enough that the fluorescent microspheres can be visualized through it. 
Type Of Material Biological samples 
Year Produced 2022 
Provided To Others? No  
Impact This is the first time, to our knowledge, that microrheological methods are applied to the study of insects. We are now obtaining the first measurements of the viscoelastic properties of fluids in live katydids. Our preliminary results show that both the hemolymph and the acoustic vesicle fluid behave as Newtonian fluids at a wide range of frequencies, and are several times more viscous than water. Furthermore, we have unequivocally shown that the acoustic vesicle of the katydids Phlugis spp and Copiphora spp., who have highly evolved hearing systems, is not linked to the circulatory system of the organism. The in situ microrehology procedure we have developed will allow not just measuring average viscoselasticity moduli but also mapping of viscosity within the organ, assessment of anisotropy and characterization of fluid dynamics in response to acoustic stimuli. The method will be described and will be available upon publication. 
 
Title Used of scaled 3D-geometries of insect ears to assess hearing capabilities and ecological interactions 
Description Since the last report for this project, a great deal of progress has been made in the geometric measurement of the ensiferan hearing system by means of micro-CT scanning and reconstruction. Micro-CT reconstruction of the acoustic tracheae, and associated measurements such as trachea length, volume, and cross-sectional area have been made for over 150 species, representing nearly 300 individual tracheae. The dataset primarily constitutes members of the Tettigoniidae, which display the clearest acoustic tracheal systems of ensiferan orthopterans, for which 12 of the 18 extant subfamilies are currently represented in the dataset. We have also recently been able to extend this dataset into the evolutionary history of the Ensifera, by means of micro-CT scanning and reconstructing the trachea of amber fossils aged from 45-40 million years old. Moving forward, extending the fossil dataset will permit greater evolutionary inferences of the biophysical adaptations of the ensiferan ear through history. As well as the acoustic tracheae, we have been able to reconstruct the external auditory pinnae of many species, which have been 3D printed for future biophysical experimentation. This recently developed methodology [Pulver at al. (doi:10.1101/2021.09.01.458595) in review in e-Life] has demonstrated that it is possible to use scaled 3D models of ensiferan auditory components to investigate hearing in ways not possible in live insects. The recent work suggests that the pinnae of neotropical tettigoniids function for enhanced detection of ultrasounds, to act as an early warning system against echolocating bats. 
Type Of Material Physiological assessment or outcome measure 
Year Produced 2021 
Provided To Others? Yes  
Impact By utilizing the extensive 3D dataset already collected for the trachea measurements, the project will be able to characterize the morphological and functional diversity of these pinnae in the very near future. At the moment we are implementing this innovative technique in amber fossils, to infer hearing ranges and other aspects of the acoustic ecology in extinct environments. Apart from the micro-CT and 3D printing costing, this technique only requires audio equipment to study ultrasonic signals (as the sound wavelength is also downscaled to match the scaled ear), therefore can be of interest to many research groups interested in ultrasound hearing, especially in those ear structures difficult to access in living animals. 
URL https://www.biorxiv.org/content/10.1101/2021.09.01.458595v4
 
Title micro-CT and 3D reconstruction of acoustic tracheal (or ear canal) diversity 
Description Micro-CT scanning of the trachea of orthopteran insects. Following removal from their storage in 80-100% ethanol, specimens are placed on dry paper towel to allow time for external ethanol to evaporate. Specimens are then held at the acoustic spiracle with a cotton q-tip to remove ethanol from within the acoustic trachea. Before the rest of the specimen dries out and ethanol leaves the soft tissues, which likely would take just under an hour in the largest species, the specimens are placed in a custom build holder for micro-CT scanning. The scans are completed at 50-55 kV, and 180-200 micro-amps, at a pixel size of 12-13 microns, with a 600 ms camera exposure and rotation steps of 0.2 degrees from 0 to 180 degrees. These settings permit a 20-30 minute scan, allowing for imaging of the acoustic tracheal system before the specimen starts to move in the scanner as it dries out. Once the scan is complete, further measurements are made such as body size and leg length, before the specimen can return to the ethanol with no damage to the sample. Following scanning, data can be segmented in Amira 9.4, to extract the geometry of the acoustic trachea. We can then measure the volume, length, and radius of the trachea, and extract the data and 3D images for the cross-species and inter-disciplinary comparison. 
Type Of Material Biological samples 
Year Produced 2021 
Provided To Others? No  
Impact We have found that we are able to obtain the geometry of the ear canal of these insects in quite old specimens, dating back to 1997 at the earliest so far. This is surprising given that the structures are made of soft tissue, but is very fortunate as it means we can take advantage of existing stored specimens to greatly enhance the size and diversity of the dataset. The 3D geometries are being used for numerical modelling, therefore such data set also will have impact in the 3Rs research as a form of reducing experimental animals. The method will be described and will be available upon publication. 
 
Title Data for: Ear pinnae in a neotropical katydid (Orthoptera: Tettigoniidae) function as ultrasound guides for bat detection 
Description Early predator detection is a key component of the predator-prey arms race and has driven the evolution of multiple animal hearing systems. Katydids (Insecta) have sophisticated ears, each consisting of paired tympana on each foreleg that receive sound both externally, through the air, and internally via a narrowing ear canal running through the leg from an acoustic spiracle on the thorax. These ears are pressure-time difference receivers capable of sensitive and accurate directional hearing across a wide frequency range. Many katydid species have cuticular pinnae which form cavities around the outer tympanal surfaces, but their function is unknown. We investigated pinnal function in the katydid Copiphora gorgonensis by combining experimental biophysics and numerical modelling using 3D ear geometries. We found that the pinnae in C. gorgonensis do not assist in directional hearing for conspecific call frequencies, but instead act as ultrasound detectors. Pinnae induced large sound pressure gains (20-30 dB) that enhanced sound detection at high ultrasonic frequencies (>60 kHz), matching the echolocation range of co-occurring insectivorous gleaning bats. These findings were supported by behavioural and neural audiograms and pinnal cavity resonances from live specimens, and comparisons with the pinnal mechanics of sympatric katydid species, which together suggest that katydid pinnae primarily evolved for the enhanced detection of predatory bats. 
Type Of Material Database/Collection of data 
Year Produced 2021 
Provided To Others? Yes  
Impact Our new methodologies for conducting scaled biophysical experimentation on 3D printed models rather than real insects realistically reduces the requirements of live insects for this research. While live animals may still be required to enhance the dataset into the future, these novel methodologies could be utilised to first search through natural history collections to see if it is possible to obtain future data in collection specimens. 
URL http://datadryad.org/stash/dataset/doi:10.5061/dryad.k0p2ngf8x
 
Description Dynamic experiments of wing motion during sound production on crickets 
Organisation University of Oxford
Department Oxford Hub
Country United Kingdom 
Sector Academic/University 
PI Contribution The aim of the proposed collaboration is to image and study how the cricket wing can be used for multiple functions - that of flight and singing. This project will leverage and further optimise the application of time-resolved X-ray microtomography as a key method for the study of complex repetitive motions in microscopic biomechanical systems, beyond those that are periodic. The PI's lab has developed a neurophysiology technique that elicit sound production in male crickets. This technique induce stridulation in crickets for long periods, allowing any dynamic experiment using X-ray in the synchrotron.
Collaborator Contribution Our parents obtained living crickets, maintained them in captivity to produce healthy adults. They also handled the two rounds of PSI project approval. They also developed a novel scanning and reconstruction protocol that allowed us to image the semi-periodic biomechanical patterns of live, singing crickets, which permits to reconstruct the muscular motion associated to sound production, in 3D.
Impact No outputs published yet, but we are waiting for the following section at the PSI to produce relevant papers.
Start Year 2022
 
Description Evidence for convergent evolution of harmonic hopping and multiple origins of high-frequency calls in crickets 
Organisation National History Museum, Paris, France
Country France 
Sector Public 
PI Contribution This collaboration is between the PI's lab and the Muséum national d'Histoire naturelle Paris (Prof. Tony Robillard and his PhD. student Teddy Gaiddon). They stud a groups of crickets (Eneopterinae) that differ from most crickets in that they evolved high frequency (HF) sound communication. The questions study in this collaborations are: 1) How are HF calls produced in Pseudolebinthus in regard to the unique features of their forewings? 2 Considering morphological, acoustical, and biomechanical lines of evidence, what is the origin of HF songs in eneopterine crickets? Did HF calling via harmonic hopping evolve once in the ancestor of a larger clade (H0) or multiple times independently within the tribes Eneopterini, Xenogryllini and Lebinthini (H1)? The PI's lab produced the Laser Doppler vibrometry to study wing resonances and well as the Micro-CT scan and 3D reconstruction of wing for further numerical modelling, to present acoustic evidence that the calls are indeed a result of harmonic frequency hopping from an ancestral low-frequency fundamental to its fourth harmonic. The PI and PDRA Charlie Woodrow advise student Teddy Gaiddon on the numerical analysis.
Collaborator Contribution Our colleagues in Paris conduct field trips to Malawi, East Africa (export permit number EAD-12-07-087-18-20a from the Forestry Research Institute of Malawi (FRIM)), maintain colonies of these insects and produce call recordings. Student Teddy Gaiddon is currently being trained in the numerical analysis process using COMSOL. They also provide morphological and biomechanical support for a mechanism explaining the observed frequency jump. Using phylogenetic analyses, they show harmonic hopping events in eneopterine crickets occurred multiple times in various acoustical and morphological contexts independently, thereby constituting an example of convergent evolution of an acoustic trait.
Impact A research paper is being prepared for the Journal of Experimental Biology.
Start Year 2022
 
Description Reconstruction of acoustic behaviour in Museum specimens of extinct and alive forms of Ensifera insects (crickets and bush crickets) 
Organisation Natural History Museum
Country United Kingdom 
Sector Public 
PI Contribution Working on reconstruction of the sound of a 150 year old insect, to better understand acoustic communication in ancient ensiferans. The species Prophalangopsis obscura is known from only a single specimen, housed at the NHM London. We have borrowed the specimen to image the sound production organs (fore wings) using our Alicona 3D surface scanning equipment, and scanned these wings using laser-doppler vibrometry. This information will be used to reconstruct the song of the species.
Collaborator Contribution Researching the history of the specimen of Prophalangopsis obscura. Trying to figure out the history of this specimen will allow for better identification of the region in which it was collected, and may still exist. This information will also form part of a small report kept in the museum's data repository for future curators interested in the specimen. In combination with results from our experiments, this will allow us to better understand the history of this elusive group of insects.
Impact A paper was published in PLoS ONE 17(8): e0270498. DOI: 10.1371/journal.pone.0270498. This collaboration is multidisciplinary operating in the interface of Evolutionary Biology, taxonomy, biophysics and engineering.
Start Year 2020
 
Description Auditory mechanics in a primitive insect 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Industry/Business
Results and Impact PDRA and PI presented an interactive poster in this event, aimed at networking between academics and the industry sector. There was over 150 people, and several gourd were interested in our approach to numerical modelling using the Finite Element Analysis method.
Year(s) Of Engagement Activity 2022
 
Description Interview for International News 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact During the press release of our journal article: Woodrow C. Baker E, Jonsson T, & Montealegre-Z F. 2022. Reviving the sound of a 150-year-old insect: the bioacoustics of Prophalangopsis obscura (Ensifera: Hagloidea). PLoS ONE 17(8): e0270498. DOI: 10.1371/journal.pone.0270498, PDRA Charlie Woodrow and first author of the paper and the PI gave an interview to the New York Times (journalist Jack Tamisiea) on the 5th of August, 2022. The interview was quickly disseminated and
Year(s) Of Engagement Activity 2022
URL https://www.nytimes.com/2022/08/10/science/singing-insect-wing-sound.html?unlocked_article_code=AAAA...
 
Description Journalists and media consultation 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Reported Dan Vergano consulted the PI to inform that the US government, though the JASON group had supported our research showing that the the AP agency sound recorded attributed to the sonic attach to the US diplomats was only a male cricket singing. He also pointed the fact that the US House of Representatives voted 427-0 to pass a "Havana Act". Mr. Vergano wanted to asked my opinion on the matter, mostly the PI personal views and feelings about of being supported in a research aspect that was contradicted before.
Year(s) Of Engagement Activity 2021
URL https://www.buzzfeednews.com/article/danvergano/havana-syndrome-jason-crickets
 
Description Live demonstrations of acoustic experiments in 3D printed insects ears 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact During the First Bioacoustics conference, PDRA Charlie Woodrow and PI were given a stand to demonstrate how hearing experiments could be conducted on scaled 3D printed geometries of insects ears. For most people this was a completely different approach to bioacoustics. Another impact was that the demonstration also provide new views to people in the use of preserved Museum specimens, which can be scanned using micro-CT, printed in 3D and used in this type of experiments. This obviously has an impact on the 3Rs.
Year(s) Of Engagement Activity 2022