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Hunter, M.

Publications and source records attributed to Hunter, M..

At least 19 recordsLinked to original sources

Growth capacity and mortality burden shape microbial persistence under osmotic stress in Bacteroides thetaiotaomicron

Microbial persistence in the gut is central to sustaining beneficial host-microbiota interactions; however, characterizing determinants of persistence in vivo can be challenging due to the complexity of native microbial communities interacting with the host. Here, we used a defined microbial consortium to demonstrate that intrinsic in vitro growth capacity becomes increasingly predictive of in vivo relative abundance as intestinal osmolality increases. In Bacteroides thetaiotaomicron, in vivo abundance was higher than predicted from growth capacity alone, indicating that additional physiological factors contribute to persistence. We identify mortality as one such determinant: adaptation to prolonged osmotic stress markedly reduced cell loss without enhancing growth capacity. This adaptive phenotype was associated with rapid and reversible phase variation in cell surface-associated PUL78/80 rather than fixed de novo mutations, with the PUL78/80-OFF state becoming progressively enriched among surviving cells as mortality increased. Our findings demonstrate that in addition to growth capacity, reduced cell loss can provide a distinct route to microbial persistence during environmental perturbation.

microbiology↗

Regional cerebral arterial elasticity is associated with cardiovascular health in cognitively healthy older adults

Ageing is often associated with a decline in cardiovascular and cerebrovascular health. This study examines the relationship between cerebral arterial elasticity and measures of cardiovascular health, as well as longitudinal changes in cerebral arterial elasticity over a 1.5 year interval. The pulse relaxation function (PReFx) is a measure of regional cerebral arterial elasticity derived using diffuse optical tomography (pulse-DOT). PReFx was measured over the anterior brain, including the frontal lobes and anterior sections of temporal and parietal regions that are especially vulnerable to vascular and cognitive ageing. We examined relationships between PReFx and measures of four cardiovascular risk factors (CVRF; i.e., hypertension, cholesterol, diabetes, obesity), as well as CVRF burden (i.e., number of CVRFs) in the highly active and cognitively healthy ACTIVate cohort (60-70 years). We replicated the well-established relationship between PReFx and both age and cardiorespiratory fitness, and examined associations between PReFx, measures of cardiovascular health and CVRF burden. Higher CVRF burden was linearly associated with lower cerebral arterial elasticity. The relationship between age and cerebral arterial elasticity was partially mediated by pulse pressure, an index of hypertension. PReFx declined significantly in as little as 1.5 years, and the effect did not vary with baseline level of any of the four CVRFs. We conclude that PReFx shows promise as a putative biomarker for monitoring cerebrovascular ageing in healthy older adults.

neuroscience↗

The effect of cell death on DNA-replication-based estimates of microbial population growth

Inferring bacterial growth rates is fundamental to understanding microbial interactions and community dynamics, but remains difficult in natural settings where time points are limited or organisms are unculturable. In these cases, a widely used method is the origin-to-terminus ratio, or peak-to-trough ratio (PTR), which estimates DNA replication activity from origin and terminus copy numbers. Although PTR is theoretically related to bacterial growth rate, it is frequently benchmarked against population growth rate, which reflects the balance between cell division and loss. Understanding when and why these measures diverge is therefore important for validating PTR-based approaches. To quantify how departures from steady growth affect PTR, population growth rate, and bacterial growth rate, we developed a stochastic, cell-based model that explicitly tracks DNA replication, cell division, and cell death. We show that transient stress-induced mortality and heterogeneous survival can cause PTR to fail to reflect population growth rate while still reflecting bacterial growth rate. We experimentally validated these predictions by exposing \textit{Escherichia coli} to osmotic shock or antibiotics, and measuring population growth rate and DNA replication activity. We further show that initial physiological state, lag before replication resumes, and abrupt growth arrest can alter the relationship between PTR and population growth rate. In the presence of mortality, lag can increase PTR's correspondence with population growth rate while decreasing its correspondence with bacterial growth rate. These results provide a mechanistic and quantitative framework for interpreting PTR across changing growth states, mortality regimes, and environmental perturbations.

microbiology↗

Orthogonal Transposons for Iterative Genome Engineering of Mammalian Cells.

The contemporary shift toward multispecific antibodies, antibody-drug conjugates (ADCs), and bespoke glycoengineered therapeutics have exposed the limitations of standard genomic engineering tools. This paper presents a novel iterative engineering paradigm utilizing the Leap-In Transposase(R) platform. By leveraging a suite of three mutually orthogonal transposase-transposon systems, we demonstrate the sequential modification of the Chinese Hamster Ovary (CHO) genome to achieve three distinct functional outcomes: (i) First, the creation of a glutamine synthetase (GS)-deficient host (CHO-K1-GS) via targeted knockdown, (ii) Second, the integration of multiple copies of a model therapeutic IgG1 for expression, and (iii) Third, the subsequent knockdown of the fucosylation pathway to modulate the glycan profile of the expressed IgG1. Genetic stability (copy number & sequence) of each integration event was confirmed using Targeted Locus Amplification (TLA) and Next-Generation Sequencing (NGS). Functional stability (expression levels, metabolic phenotype, and glycan phenotypes) was confirmed using standard cell culture and analytical techniques. Crucially, the truly orthogonal nature of the transposase-transposon pairs prevents cross-mobilization and ensures the structural and functional integrity of previously integrated cargo. This study establishes a "What You See Is What You Get" (WYSIWYG) methodology that provides a robust, scalable, and predictable framework for developing next-generation complex biopharmaceutical manufacturing cell lines.

bioengineering↗

Unveiling the taxonomic diversity and unprecedented biosynthetic treasure of the phylum Myxococcota

Natural products remain vital sources of therapeutics, particularly anti-infectives, and members of the phylum Myxococcota constitute an especially rich reservoir for their discovery. Based on decades of microbiological efforts, we present 154 new Myxococcota genomes and propose a revised taxonomy expanding the number of described families from 11 to 28 and genera from 32 to 90. Comparison with an equivalent set from the prime source Actinomycetota shows that Myxococcota possess a comparable biosynthetic diversity, underscoring their promise for large-scale isolation and sequencing efforts. The vast untapped potential reflected in 2,387 uncharacterized gene cluster families is highlighted by genome mining efforts, yielding four validated compounds exhibiting novel chemistry, including myxolutamids and myxopentacins. We show that many Myxococcota-derived natural products, such as myxolutamid A and two new sorangicin derivatives, are conserved within taxonomic lineages. New described families thus bear high biosynthetic potential underpinning the importance of precise taxonomic classification guiding targeted drug discovery.

microbiology↗

Cell Line Development for Bispecific Antibodies: Better Predictability Through Transposases

Bispecific antibodies are at the forefront of biopharmaceutical drug development. With over 100 different molecular architectures combined with diverse individual subunit sequences, choosing the most suitable structure and predicting the ideal subunit expression ratios for successful heterodimerization is a significant challenge. In this paper, we demonstrate that the recently described cell line development paradigm shift (Rajendran et al. 2021), enabled by the Leap-In transposon platform, can be extended to the development of bispecific monoclonal antibody-producing cell substrates (stable clones and pools). The key features are 1) Parental pools reliably predict the derivative clonal productivity and clonal heterodimer fractions. 2) Clonal productivity and clonal heterodimer fraction remained stable for at least 60 population doublings. 3) Depending on the products biophysicochemical properties, the stable pools exhibit variable productivity stability. 4) Heterodimer fractions remain stable in the Leap-In mediated stable pools independently of the productivity stability of the pools. 5) Structures and subunit ratios can be triaged at stable pool level, and 6) Due to the homogeneous clonal productivity distribution, only a small number ([~]50) of clones need to be isolated and characterized.

bioengineering↗

Live and let die: lysis time variability and resource limitation shape lytic bacteriophage fitness

Bacteriophages (phages) play a critical role in controlling bacterial populations, both in nature and as potential therapeutic agents. Their ability to replicate, compete against each other, and eradicate target cell populations is usually understood through a number of life history parameters, traditionally measured by population-level assays, which implicitly average the parameters value across a large number of infection events. Recent experiments suggest that bacteriophage life history parameters are subject to considerable heterogeneity, raising the question of whether experimental and modelling efforts that do not account for this variability may overlook important factors in phages behaviour, competitive fitness or therapeutic viability. Here, using agent-based simulations, we investigate the importance of stochasticity in lysis time and burst size of lytic bacteriophages in two common laboratory competition experiments: serial passage of well-mixed populations and plaque expansion across a bacterial lawn. We find that a phages analytic growth rate in isolation can be a poor predictor of its fitness advantage in simulated competition experiments. Specifically, when lysis times are tightly distributed, we identify a novel effect we name "population resonance", through which a bacteriophage can display a significant fitness advantage over a competitor with a much greater growth rate in isolation. Our simulations also show that both serial passage and plaque expansion reward variability in lysis time more than expected, by increasing the phage resilience when resources are scarce. Author SummaryBacteriophages (viruses that infect bacteria) can be described by a set of attributes called life history parameters. Historically, these parameters could only be measured on-average over large populations. Recent experimental advancements, however, have enabled their quantification at the single-cell-single-virus level, and have revealed that these traits are subject to inherent variability, even for genetically identical viruses infecting genetically identical cells. Here, we explore whether the degree of variability in one particular life history parameter, the lysis time (the time taken for a bacteriophage to infect and kill its host bacteria) might be subject to natural selection, by quantifying its effects on phages fitness in two simulated competition experiments: serial passage and plaque expansion. Surprisingly, we find that lysis time variability is advantageous in competition even when it reduces the phages growth rate in isolation. This finding opens the door to future computational and experimental work, as it demonstrates that mean values alone are not sufficient to describe or predict a bacteriophage efficacy, and that models that ignore this variability can overlook viable evolutionary strategies.

evolutionary biology↗

Cardiorespiratory fitness and cardiometabolic health are associated with distinct cognitive domains in cognitively healthy older adults.

BackgroundAging is associated with progressive cognitive decline, as well as increased prevalence of cardiometabolic risk factors and reduced cardiorespiratory fitness. In fact, reduced cardiometabolic health and cardiorespiratory fitness are both associated with a decline in cognitive functioning. This study examines the common and distinct contributions of these cardiovascular health factors on cognitive variability across different domains in cognitively healthy older adults. Methods and ResultsWe apply structural equation modelling (SEM) to model cross-sectional relationships between cardiometabolic health, cardiorespiratory fitness and performance across multiple cognitive domains in an age-restricted sample of healthy older adults from the ACTIVate Study (n=345; 60-70 yrs). Participants completed a series of cognitive and clinical assessments (including brachial blood pressure, heart rate, blood-based metabolic markers). We designed a cognitive model (Model 1) with four latent factors that are differentially impacted by aging (Processing Speed, Executive Function, Verbal Memory and Crystallized Ability) and used it to test effects on cognition of two theory-driven dimensions of cardiovascular health: Cardiorespiratory Fitness (Model 2) and Cardiometabolic Health (Model 3). Model 4 included both predictors and examined their joint and distinct effects on these cognitive domains. When controlling for their joint variance, Cardiometabolic Health and Cardiorespiratory Fitness showed evidence consistent with a double dissociation on cognitive domains. Specifically, cardiorespiratory fitness significantly predicted processing speed (r=0.28, p<0.05) and executive function (r=0.66, p<0.05), but not verbal memory and crystallized ability. In contrast, cardiometabolic health predicted crystallized ability (r=0.31, p<0.05) and verbal memory (r=0.28, p<0.05), but not executive function and processing speed. ConclusionsThis study shows the first evidence that cardiorespiratory fitness and cardiometabolic health are associated with distinct cognitive domains in a large cross-sectional, age-restricted and high functioning cohort. These findings emphasize the importance of healthy aging approaches that target both health literacy and lifestyle behaviors to promote functional capacity across the lifespan.

neuroscience↗

Osmotic conditions shape fitness gains and resistance mechanisms during E. coli and T4 phage co-evolution

Environmental conditions strongly influence interactions between bacteria and bacterio-phages (phages). Here, we examined how osmolality (solute concentration) shapes the in vitro co-evolution of T4 phage and its host Escherichia coli during serial passage. When evolved independently, we observed substantial fitness gains in both bacteria and phages, particularly in high osmotic conditions. During co-evolution, however, fitness gains were limited, bacterial populations consistently evolved phage resistance, and several phage populations went extinct. Furthermore, the resistance mechanisms varied by osmolality. In lower osmolalities, mutations disrupted phage binding sites, conferring strong resistance. In higher osmolalities, mutations led to increased colonic acid production, producing a mucoid phenotype with weaker resistance. Because mucoidy has been associated with increased bacterial virulence, these findings suggest that gut-relevant osmotic conditions may constrain evolutionary trajectories, favoring resistance strategies that are less effective against phage but potentially more virulent, with important implications for phage therapy design. SignificancePhages offer a promising alternative to antibiotics, but their safety and efficacy strongly depends on the environmental conditions where the bacteria and phages interact. In the human gut, for instance, solute concentrations can vary widely due to factors like food intolerances or laxative use. In this study, we show that such variations significantly impact how bacteria and phages co-evolve. In particular, we find that in higher osmolalities, bacteria evolve phage resistance through mucoidy - a phenotype linked with increased bacterial virulence - rather than receptor loss. This highlights the need to consider environmental factors when developing phage therapies.

microbiology↗

Real time imaging reveals a role for macrophage protrusive motility in melanoma invasion

Macrophages are primary cells of the innate immune system that mediate tumor progression. However, the motile behavior of macrophages and interactions with tumor cells are not well understood. Here, we exploit the optical transparency of larval zebrafish and perform real time imaging of macrophage-melanoma interactions. We found that macrophages are highly motile in the tumor microenvironment. Macrophages extend dynamic projections between tumor cells that precedes invasive melanoma migration. Modulating macrophage motility with a dominant inhibitory mutation in Rac2 inhibits recruitment to the tumor and impairs tumor invasion. However, a hyperactivating mutation in Rac2 does not affect macrophage recruitment but limits macrophage projections into the melanoma mass and reduces invasive melanoma cell migration. Taken together, these findings reveal a role for Rac2-mediated macrophage protrusive motility in melanoma invasion.

cell biology↗

Single-cell imaging of the lytic phage life cycle dissects the timing and noise in individual infection steps

When a lytic bacteriophage infects a bacterial cell, it commandeers the cells resources to replicate, ultimately causing cell lysis and the release of new virions. As phages function as obligate parasites, each stage of the infection process depends on the physiological parameters of the host cell. Given the inherent physiological variability within a population of genetically identical bacterial cells, we ask how the phage infection dynamic reflects such heterogeneity. Here, we introduce a timelapse imaging assay for investigating the dynamics of individual infection steps by a single T7 phage on a single bacterium. This high-throughput, time-resolved assay enables us to monitor the infection progression simultaneously in multiple cells, uncovering substantial heterogeneity at each step and revealing correlations between infection dynamics and the physiological state of the infected cell. Simulations of competing phage populations with different lysis time distributions reveal that heterogeneity in infection dynamics can significantly impact phage fitness, highlighting it as a potential evolutionary driver of phage-bacteria interactions.

microbiology↗

Heterogeneity in the M. tuberculosis β-lactamase inhibition by Sulbactam

For decades, researchers have been determined to elucidate essential enzymatic functions on the atomic lengths scale by tracing atomic positions in real time. Our work builds on new possibilities unleashed by mix-and-inject serial crystallography (MISC)1-5 at X-ray free electron laser facilities. In this approach, enzymatic reactions are triggered by mixing substrate or ligand solutions with enzyme microcrystals6. Here, we report in atomic detail and with millisecond time-resolution how the Mycobacterium tuberculosis enzyme BlaC is inhibited by sulbactam (SUB). Our results reveal ligand binding heterogeneity, ligand gating7-9, cooperativity, induced fit10,11 and conformational selection11-13 all from the same set of MISC data, detailing how SUB approaches the catalytic clefts and binds to the enzyme non-covalently before reacting to a trans-enamine. This was made possible in part by the application of the singular value decomposition14 to the MISC data using a newly developed program that remains functional even if unit cell parameters change during the reaction.

molecular biology↗

Genomic and ecologic characteristics of the airway microbial-mucosal complex

SO_SCPLOWUMMARYC_SCPLOWO_SCPCAP C_SCPCAPO_SCPLOWPARAGRAPHC_SCPLOWLung diseases due to infection and dysbiosis affect hundreds of millions of people world-wide1-4. Microbial communities at the airway mucosal barrier are conserved and highly ordered5, reflecting symbiosis and co-evolution with human host factors6. Freed of selection to digest nutrients for the host, the airway microbiome underpins cognate management of mucosal immunity and pathogen resistance. We show here the results of the first systematic culture and whole-genome sequencing of the principal airway bacterial species, identifying abundant novel organisms within the genera Streptococcus, Pauljensenia, Neisseria and Gemella. Bacterial genomes were enriched for genes encoding antimicrobial synthesis, adhesion and biofilm formation, immune modulation, iron utilisation, nitrous oxide (NO) metabolism and sphingolipid signalling. RNA-targeting CRISPR elements in some taxa suggest the potential to prevent or treat specific viral infections. Homologues of human RO60 present in Neisseria spp. provide a possible respiratory primer for autoimmunity in systemic lupus erythematosus (SLE) and Sjogren syndrome. We interpret the structure and biogeography of airway microbial communities from clinical surveys in the context of whole-genome content, identifying features of airway dysbiosis that may presage breakdown of homeostasis during acute attacks of asthma and chronic obstructive pulmonary disease (COPD). We match the gene content of isolates to human transcripts and metabolites expressed late in airway epithelial differentiation, identifying pathways that can sustain host interactions with the microbiota. Our results provide a systematic basis for decrypting interactions between commensals, pathogens, and mucosal immunity in lung diseases of global significance.

microbiology↗

Rational control of structural off-state heterogeneity in a photoswitchable fluorescent protein provides switching contrast enhancement

Reversibly photoswitchable fluorescent proteins are essential markers for advanced biological imaging, and optimization of their photophysical properties underlies improved performance and novel applications. Here we establish a link between photoswitching contrast, a key parameter that largely dictates the achievable resolution in nanoscopy applications, and chromophore conformation in the non-fluorescent state of rsEGFP2, a widely employed label in REversible Saturable OpticaL Fluorescence Transitions (RESOLFT) microscopy. Upon illumination, the cis chromophore of rsEGFP2 isomerizes to two distinct off-state conformations, trans1 and trans2, located on either side of the V151 side chain. Reducing or enlarging the side chain at this position (V151A and V151L variants) leads to single off-state conformations that exhibit higher and lower switching contrast, respectively, compared to the rsEGFP2 parent. The combination of structural information obtained by serial femtosecond crystallography with high-level quantum chemical calculations and with spectroscopic and photophysical data determined in vitro suggests that the changes in switching contrast arise from blue- and red-shifts of the absorption bands associated to trans1 and trans2, respectively. Thus, due to elimination of trans2, the V151A variants of rsEGFP2 and its superfolding variant rsFolder2 display a more than two-fold higher switching contrast than their respective parent proteins, both in vitro and in E. coli cells. The application of the rsFolder2-V151A variant is demonstrated in RESOLFT nanoscopy. Our study rationalizes the connection between structural and photophysical chromophore properties and suggests a means to rationally improve fluorescent proteins for nanoscopy applications.

biophysics↗

Population-specific patterns of toxin sequestration in monarch butterflies from around the world

Animals frequently defend themselves against predators using diet-derived toxins. Monarch butterflies are a preeminent example of toxin sequestration, gaining protection via cardenolides in their milkweed hosts. Few studies have considered genetic variation in sequestration ability, in monarchs or other species. Here, we use two approaches to study natural selection on cardenolide sequestration in monarchs. First, we conducted a reciprocal rearing experiment with six monarch populations and six associated host species from around the world to determine whether sequestration is higher in monarchs reared on sympatric host species. Second, we compared sequestered cardenolides in wild-caught monarchs from Guam--an island where bird predators have been functionally extirpated for >40 years--to a nearby island with intact birds. We found substantial genetic variation in sequestration ability, though no consistent sequestration advantage in sympatric combinations. One monarch population from Puerto Rico showed greatly reduced sequestration from Asclepias syriaca, likely reflecting a lack of evolutionary association with this host. Monarchs from Guam showed reduced sequestration from A. curassavica, both in a cross-island comparison and when reared under controlled conditions. Our results suggest that processes involved in toxin sequestration are subject to natural selection and may evolve in response to contemporary changes in species interactions.

evolutionary biology↗

Preventing co-infection: a viral strategy with short-term benefits and long-term drawbacks

Viral superinfection occurs when multiple viral particles subsequently infect the same host. In nature, several viral species are found to have evolved diverse mechanisms to prevent superinfection (superinfection exclusion) but how this strategic choice impacts the fate of mutations in the viral population remains unclear. Using stochastic simulations, we find that genetic drift is suppressed when superinfection occurs, thus facilitating the fixation of beneficial mutations and the removal of deleterious ones. Interestingly, we also find that the competitive (dis)advantage associated with variations in life history parameters is not necessarily captured by the viral growth rate for either infection strategy. Putting these together, we then show that a mutant with superinfection exclusion will easily overtake a superinfecting population even if the latter has a much higher growth rate. Our findings suggest that while superinfection exclusion can negatively impact the long-term adaptation of a viral population, in the short-term it is ultimately a winning strategy. Author summaryViral social behaviour has recently been receiving increasing attention in the context of ecological and evolutionary dynamics of viral populations. One fascinating and still relatively poorly understood example is superinfection or co-infection, which occur when multiple viruses infect the same host. Among bacteriophages, a wide range of mechanisms have been discovered that enable phage to prevent superinfection (superinfection exclusion) even at the cost of using precious resources for this purpose. What is the evolutionary impact of this strategic choice and why do so many phages exhibit this behaviour? Here, we conduct an extensive simulation study of a phage population to address this question. In particular, we investigate the fate of viral mutations arising in an environment with a constant supply of bacterial hosts designed to mimic a "turbidostat," as these are increasingly being used in laboratory evolution experiments. Our results show that allowing superinfection in the long-term yields a population which is more capable of adapting to changes in the environment. However, when in direct competition, mutants capable of preventing superinfection experience a very large advantage over their superinfecting counterparts, even if this ability comes at a significant cost to their growth rate. This indicates that while preventing superinfection can negatively impact the long-term prospects of a viral population, in the short-term it is ultimately a winning strategy.

evolutionary biology↗

Virus-host interactions shape viral dispersal giving rise to distinct classes of travelling waves in spatial expansions

Reaction-diffusion waves have long been used to describe the growth and spread of populations undergoing a spatial range expansion. Such waves are generally classed as either pulled, where the dynamics are driven by the very tip of the front and stochastic fluctuations are high, or pushed, where cooperation in growth or dispersal results in a bulk-driven wave in which fluctuations are suppressed. These concepts have been well studied experimentally in populations where the cooperation leads to a density-dependent growth rate. By contrast, relatively little is known about experimental populations that exhibit density-dependent dispersal. Using bacteriophage T7 as a test organism, we present novel experimental measurements that demonstrate that the diffusion of phage T7, in a lawn of host E. coli, is hindered by steric interactions with host bacteria cells. The coupling between host density, phage dispersal and cell lysis caused by viral infection results in an effective density-dependent diffusion coefficient akin to cooperative behavior. Using a system of reaction-diffusion equations, we show that this effect can result in a transition from a pulled to pushed expansion. Moreover, we find that a second, independent density-dependent effect on phage dispersal spontaneously emerges as a result of the viral incubation period, during which phage is trapped inside the host unable to disperse. Additional stochastic agent-based simulations reveal that lysis time dramatically affects the rate of diversity loss in viral expansions. Taken together, our results indicate both that bacteriophage can be used as a controllable laboratory population to investigate the impact of density-dependent dispersal on evolution, and that the genetic diversity and adaptability of expanding viral populations could be much greater than is currently assumed.

biophysics↗

Reporting to the Convention on Biology Diversity: How are countries assessing and protecting their genetic diversity?

Genetic diversity is critically important for all species-domesticated and wild- to adapt to environmental change, and for ecosystem resilience to extreme events. International agreements such as the Convention on Biological Diversity (CBD) have committed to conserve and sustainably and equitably use all levels of biodiversity-genes, species and ecosystems-globally. However, assessment and monitoring of genetic diversity are often overlooked, and there are large knowledge and policy gaps regarding genetic diversity conservation. In this study, we present the first quantitative analysis of genetic diversity assessments conducted by Parties to the CBD. We conducted a detailed, systematic analysis of 114 CBD 5th (submitted 2014) and 6th (submitted 2018) National Reports to quantitatively assess actions, progress on targets, values and indicators related to genetic diversity. First, we found that the importance of genetic diversity is recognised by most Parties to the CBD, and that recognition increased over time. However, genetic targets mainly addressed genetic diversity within cultivated plants, farm animals, and crop wild relatives, with little focus on other wild species. Also, actions for conserving genetic diversity primarily concerned ex-situ facilities and policy, rather than monitoring and intervention for maintaining genetic diversity in situ. The most commonly used indicators of genetic diversity status were the number of genetic resources in conservation facilities, number of threatened breeds, and Red List Index, which are not well correlated to genetic erosion in most species -- highlighting that genetic change is poorly monitored by current indicators. Lastly, analyses of genetic data observations, indigenous use and knowledge of genetic diversity, and strategies being developed and implemented to conserve genetic diversity are highly under-reported. We make several recommendations for the post-2020 CBD Biodiversity Framework to improve awareness, assessment, and monitoring, and facilitate consistent and complete reporting of progress of genetic diversity in future National Reports. Article Impact StatementAn analysis of genetic diversity in CBD National Reports neglects non-domesticated species and demonstrates need for sufficient indicators.

scientific communication and education↗