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Raymond, B.

Publications and source records attributed to Raymond, B..

15 recordsLinked to original sources

Antimicrobial resistance in a warmer world: selection and transfer of kanamycin resistance is temperature dependent in E. coli.

Climate warming consistently correlates with increased antimicrobial resistance (AMR). Understanding the biological mechanisms underpinning this relationship is important for both prediction and mitigation of the future burden of antibiotic resistance in a warmer world, especially given AMR can spread through both selection and horizontal gene transfer. Here, we experimentally measure how temperature alters susceptibility to kanamycin, selection and spread of resistance in Escherichia coli MG1655 carrying the natural plasmids R1 and RP4. We predict that susceptibility should be lowest - and plasmid transfer highest - at maximal growth rate. Thermal performance of bacterial growth peaked at 40{degrees}C and we detected a small plasmid cost - for RP4, but not R1 - in the absence of kanamycin. The thermal performance of plasmid transfer did not mirror that of bacterial growth, instead, R1 transfer peaked at 35{degrees}C with a much narrower thermal tolerance. Susceptibility to kanamycin decreased with increasing growth rate until 31{degrees}C, before increasing again as temperatures approached and exceeded optimal growth temperature. Increasing susceptibility coupled with high growth rates meant that selection was strongest at the highest measured temperature, but the shape of the relationship changed with kanamycin concentration. In mixed populations of resistant and susceptible bacteria, we found that AMR spread most where selection was strongest, irrespective of plasmid transfer. Our findings suggest increased AMR at higher temperatures is predominantly driven by higher growth rates and increased susceptibility, resulting in stronger selection. Our study provides a framework to uncover generalities across antibiotic classes, strains, species, and plasmids, laying the foundation for improved prediction of AMR selection in the environment.

microbiology↗

The insecticidal potential of Bacillus cereus group strains from insect-dense regions of the United Kingdom

Entomopathogenic Bacillus cereus group members (e.g. B. thuringiensis) are arguably the most important source of biopesticides globally and are widely used for industrial-scale crop protection. However, the application of biopesticidal B. cereus group strains is currently threatened by increasing resistance among insects, necessitating the development of novel insecticides. Here, we used whole-genome sequencing to characterize 13 novel B. cereus group strains from sites with high-density insect populations, including six isolates from an insect cadaver. Overall, diverse B. cereus group members were observed, encompassing three panC groups (II, IV and VI) and seven sequence types. Five strains possessed insecticidal Cry toxin-encoding genes with predicted target ranges that included insect species present at the respective sampling locations. Moreover, the genomes were computationally mined for biosynthetic gene clusters (BGCs; clusters of genes governing secondary metabolite synthesis). In total, 274 BGCs were observed, of which the majority (170 BGCs; 62.04%) did not cluster with known BGCs. Taken together, our study indicates that sampling B. cereus group bacteria from insect-rich ecological niches may yield Cry toxins against the self-same insects, as well as novel and unexplored BGCs, suggesting that targeted sampling efforts could generate compound leads for biopesticide development.

microbiology↗

Effect of the microbiome on pathogen susceptibility across four Drosophilidae species

The microbiota has been shown to play an important role in host susceptibility to infections in some hosts. However, less is known about whether microbiota-mediated effects are consistent across host species, as our understanding of such interactions may be affected by publication bias. Following on from a large study of 36 species of Drosophilidae challenged with four bacterial pathogens, we identified two candidate host species that might have protective microbiome based on low susceptibility and high abundance of the culturable microbiota; we selected two other host species for comparison. We tested whether germ-reduced flies, and flies with a natural or re-constituted microbiome, varied in their susceptibility to systemic infection with two bacterial pathogens (Providencia rettgeri and Staphylococcus aureus) and one viral pathogen (Drosophila C Virus). The composition and abundance of the bacterial microbiota varied between species and microbiome treatments. We found an overall interaction between host species and pathogen type, confirming previous work that host species vary in their susceptibility in a pathogen-specific manner. Similarly, we found that microbiome treatments had differing effects on host survival among host species, although effect sizes tended to be small. In D. putrida individuals with manipulated microbiomes showed increased susceptibility to all pathogens tested; in other hosts altered susceptibility was pathogen dependent. While there are always challenges to manipulating microbiomes, especially across multiple host species, our results indicate that host microbiota may play limited roles on survival in systemic infection in these four species. This work demonstrates that caution is required when generalising about the potential beneficial impact of microbiomes.

microbiology↗

Varying phylogenetic signal to four bacterial pathogens across species of Drosophilidae

Bacterial infections are one of the major sources threatening public health. Pathogen host shifts - where a pathogen jumps from one host species to another - are important sources of emerging infectious diseases. However, compared to viruses, we know relatively little about the factors that determine whether a bacteria can infect a novel host, such as how host phylogenetics constrains variation in pathogen-host range and the link between host phylogeny and the infectivity and virulence of a pathogen. Here, we experimentally examined susceptibility to bacterial infections using a panel of 36 Drosophilidae species and four pathogens (Providencia rettgeri, Pseudomonas entomophila, Enterococcus faecalis, Staphylococcus aureus). The outcomes of infection differed greatly among pathogens and across host species. The host phylogeny explains a considerable amount of variation in susceptibility, with the greatest phylogenetic signal for P. rettgeri infection, explaining 94% of the variation in mortality. Positive correlations were observed between mortality and bacterial load for three out of the four pathogens. Correlations in susceptibility between the four pathogens were positive but largely non-significant, suggesting susceptibility is mostly pathogen-specific. These results suggest that susceptibility to bacterial pathogens may be predicted by the host phylogeny, but the effect may vary in magnitude between different bacteria.

evolutionary biology↗

UNSEG: unsupervised segmentation of cells and their nuclei in complex tissue samples

Multiplexed imaging technologies have made it possible to interrogate complex tumor microenvironments at sub-cellular resolution within their native spatial context. However, proper quantification of this complexity requires the ability to easily and accurately segment cells into their sub-cellular compartments. Within the supervised learning paradigm, deep learning based segmentation methods demonstrating human level performance have emerged. However, limited work has been done in developing such generalist methods within the label-free unsupervised context. Here we present an unsupervised segmentation (UNSEG) method that achieves deep learning level performance without requiring any training data. UNSEG leverages a Bayesian-like framework and the specificity of nucleus and cell membrane markers to construct an a posteriori probability estimate of each pixel belonging to the nucleus, cell membrane, or background. It uses this estimate to segment each cell into its nuclear and cell-membrane compartments. We show that UNSEG is more internally consistent and better at generalizing to the complexity of tissue morphology than current deep learning methods. This allows UNSEG to unambiguously identify the cytoplasmic compartment of a cell, which we employ to demonstrate its use in an exemplar biological scenario. Within the UNSEG framework, we also introduce a new perturbed watershed algorithm capable of stably and automatically segmenting a cluster of cell nuclei into individual cell nuclei that increases the accuracy of classical watershed. Perturbed watershed can also be used as a standalone algorithm that researchers can incorporate within their supervised or unsupervised learning approaches to extend classical watershed, particularly in the multiplexed imaging context. Finally, as part of developing UNSEG, we have generated a high-quality annotated gastrointestinal tissue (GIT) dataset, which we anticipate will be useful for the broader research community. We demonstrate the efficacy of UNSEG on the GIT dataset, publicly available datasets, and on a range of practical scenarios. In these contexts, we also discuss the possibility of bias inherent in quantification of segmentation accuracy based on F1 score. Segmentation, despite its long antecedents, remains a challenging problem, particularly in the context of tissue samples. UNSEG, an easy-to-use algorithm, provides an unsupervised approach to overcome this bottleneck, and as we discuss, can help improve deep learning based segmentation methods by providing a bridge between unsupervised and supervised learning paradigms.

bioinformatics↗

Swimming with the current improves juvenile survival in southern elephant seals

Understanding juvenile survival is crucial for the population ecology of long-lived species, where parental guidance can significantly influence survival rates of completely naive juveniles. In southern elephant seals (Mirounga leonina), however, offspring receive no knowledge from their parents, only fat reserves. This research focuses on how dispersal direction on their maiden foraging trip and physical traits influence the survival of naive southern elephant seal pups at Macquarie Island. We tracked 44 pups with satellite tags during their post-weaning migration and compared their movements to the post-moult winter migrations of 58 adult females. We found that most pups (61.2%) travelled southeast, in line with the Antarctic Circumpolar Current. Pups travelling with the predominant east-southeast current had a 1.5 times higher survival rate for their first trip than those swimming westward against it. Those that swam with the current and were heavier were more likely to survive their first year. Adult females showed different dispersal patterns, where they travelled southwards towards Antarctic waters, implying that learning from experience influences their direction. Future investigations into the role of the primary eastward current in the sub-Antarctic on circumpolar movement patterns of top marine predators could expand our understanding of Southern Ocean ecology.

ecology↗

Manipulating multi-level selection in a fungal entomopathogen reveals social conflicts and a method for improving biocontrol traits

Changes in parasite virulence are commonly expected to lead to trade-offs in other life history traits that can affect fitness. Understanding these trade-offs is particularly important if we want to manipulate the virulence of microbial biological control agents. Theoretically, selection across different spatial scales, i.e. between- and within-hosts, shapes these trade- offs. However, trade-offs are also dependent on parasite biology. Despite their applied importance the evolution of virulence in fungal parasites is poorly understood: virulence can be unstable in culture and commonly fails to increase in simple passage experiments. We hypothesized that manipulating selection intensity at different scales would reveal virulence trade-offs in a fungal pathogen of aphids, Akanthomyces muscarius. Starting with a genetically diverse stock we selected for infectivity, parasite yield or speed of kill by manipulating competition within and between hosts and between groups of hosts over 7 rounds of infection. We characterized ancestral and evolved lineages by whole genome sequencing and by measuring virulence, growth rate, sporulation and fitness. While several lineages showed increases in virulence, we saw none of the trade-offs commonly found in obligately-killing parasites. Phenotypically similar lineages within treatments often shared multiple single-nucleotide variants, indicating strong convergent evolution. The most dramatic phenotypic changes were in timing of sporulation and spore production in vitro. We found that early sporulation led to reduced competitive fitness but could increase yield of spores on media, a trade-off characteristic of social conflict. Notably, the selection regime with strongest between-group competition and lowest genetic diversity produced the most consistent shift to early sporulation, as predicted by social evolution theory. Mutli-level selection therefore revealed social interactions novel to fungi and showed that these biocontrol agents have the genomic flexibility to improve multiple traits -virulence and spore production - that are often in conflict in other parasites. Author summaryUnderstanding the ecological forces that shape virulence is a key challenge in evolutionary biology. Here we investigated how competition at different levels of selection (within-hosts, between-hosts, between populations) could alter investment in virulence in a fungal entomopathogen. We predicted that cooperative investment in virulence would increase at higher scales of competition and aimed to further our understanding of potential trade-offs shaping life-history of a fungal insect pathogens. We found moderate increases in virulence in different selection regimes but importantly, none of the commonly expected trade-offs, such as that between spore production and virulence or a relationship between virulence and growth rate, which is consistent with cooperation. However, we found convergent genetic changes and significant differences in timing and production of spores, dependent how we manipulated scales of selection. Our data suggests that this is driven by social conflict regarding the timing of sporulation. This carries fundamental importance for understanding how varying selection pressure at different scales shape pathogen life history. In addition, these results also have applied importance for understanding how to improve and select for beneficial traits in biocontrol agents.

evolutionary biology↗

Re-inventing pathogen passage for social microbes

Passage experiments that sequentially infect hosts with parasites have long been used to manipulate virulence. However, in many invertebrate pathogens passage has been applied naively without a full theoretical understanding of how best to select for increased virulence. This has led to very mixed results. Understanding the evolution of virulence is complex because selection on parasites occurs across multiple spatial scales with potentially different conflicts operating on parasites with different life-histories. For example, in social microbes, strong selection on replication rate within hosts can lead to cheating and loss of virulence, because investment in public goods virulence reduces replication rate. In contrast, selection acting at a between host scale maintains virulence by selecting on parasite population size. In this study we tested how different scales of selection and varying mutation supply affect evolution of virulence against resistant hosts in the specialist insect pathogen Bacillus thuringiensis., aiming to optimize methods for strain improvement against a difficult to kill insect target. We show that selection for infectivity using competition between sub-populations in a metapopulation prevents social cheating, acts to retain key virulence plasmids and facilitates increased virulence. Increased virulence was associated with reduced efficiency of sporulation, and loss of function in putative regulatory genes but not with altered expression of known virulence factors. Selection in a metapopulation provides a broadly applicable tool for improving the efficacy of biocontrol agents. Moreover, a structured host population can facilitate artificial selection on infectivity, while selection on life history traits such as faster replication or larger population sizes can reduce virulence can reduce virulence in social microbes.

evolutionary biology↗

Laboratory sprayer for testing of microbial biocontrol agents: design and calibration

The lack of commercially available low-cost laboratory spraying equipment for testing microbial control agents can hinder advancement in the field of biocontrol. This study presents an inexpensive, portable sprayer that is calibrated utilizing laboratory consumables. The computer aided design files are made available so that it is freely modifiable and can be used for machine routing or 3D printing. Bioassay data was obtained by spraying Myzus persicae with spores of entomopathogenic fungi. Observed variation in droplet deposition within tested pressure and volume settings, and spore deposition within sprayed concentrations were low. Bioassay results show reproducible mortality for the tested doses.

microbiology↗

Using prior information from humans to prioritize genes and gene-associated variants for complex traits in livestock

Genome-Wide Association Studies (GWAS) in large human cohorts have identified thousands of loci associated with complex traits and diseases. For identifying the genes and gene-associated variants that underlie complex traits in livestock, especially where sample sizes are limiting, it may help to integrate the results of GWAS for equivalent traits in humans as prior information. In this study, we sought to investigate the usefulness of results from a GWAS on human height as prior information for identifying the genes and gene-associated variants that affect stature in cattle, using GWAS summary data on samples sizes of 700,000 and 58,265 for humans and cattle, respectively. Using Fishers exact test, we observed a significant proportion of cattle stature-associated genes (30/77) that are also associated with human height (odds ratio = 5.1, p = 3.1e-10). Result of randomized sampling tests showed that cattle orthologs of human height-associated genes, hereafter referred to as candidate genes (C-genes), were more enriched for cattle stature GWAS signals than random samples of genes in the cattle genome (p=0.01). Randomly sampled SNPs within the C-genes also tend to explain more genetic variance for cattle stature (up to 13.2%) than randomly sampled SNPs within random cattle genes (p=0.09). The most significant SNPs from a cattle GWAS for stature within the C-genes did not explain more genetic variance for cattle stature than the most significant SNPs within random cattle genes (p=0.87). Altogether, our findings support previous studies that suggest a similarity in the genetic regulation of height across mammalian species. However, with the availability of a powerful GWAS for stature that combined data from 8 cattle breeds, prior information from human-height GWAS does not seem to provide any additional benefit with respect to the identification of genes and gene-associated variants that affect stature in cattle. Author SummaryProvided that there is a similarity in the biological control of complex traits in different species, the results of genome-wide association study for a given trait in a model species can be utilized as prior information to identify genes and gene-associated variants that affect the same trait in a non-model species. Here we report that the number of genes found to be affecting height in both humans and cattle is more than can be expected by chance, suggesting a similarity in the biology of height in both species. In addition, genes that are associated with height in humans were found to be highly enriched with variants that are strongly associated with height in cattle. Using height as a model trait, we demonstrated that across-species prior information cannot compete with a within-species prior information, with respect to the identification of genes and the gene-associated variants that affect a complex trait. Across-species prior information can be useful, however, when within-species prior information is either unavailable or poor.

genetics↗

Optimal response to quorum-sensing signals varies in different host environments with different pathogen group size

The persistence of genetic variation in master regulators of gene expression, such as quorum-sensing systems, is hard to explain. Here, we investigated two alternative hypotheses for the prevalence of polymorphic quorum-sensing in Gram-positive bacteria, i.e. the use of different signal / receptor pairs ( pherotypes) to regulate the same functions. First, social interactions between pherotypes or facultative cheating may favour rare variants that exploit the signals of others. Second, different pherotypes may increase fitness in different environments. We evaluated these hypotheses in the invertebrate pathogen Bacillus thuringiensis, using three pherotypes expressed in a common genetic background. Facultative cheating occurred in homogenized hosts, in contrast, rare pherotypes had reduced fitness in naturalistic infections. There was clear support for environment-dependent fitness: pherotypes varied in responsiveness to signals and in mean competitive fitness. Notably, competitive fitness varied with group size: the pherotype with highest responsiveness to signals performed best in smaller hosts where infections have a lower pathogen group size. Less responsive pherotypes performed best in larger hosts. Results using homogenized insect media fit with the expectation of facultative cheating and social evolution theory, but results from naturalist oral infections do not fit many of the predictions from this body of theory. In this system, low signal abundance appears to limit fitness in hosts while the optimal level of response to signals varies in different host environments.\n\nImportanceQuorum sensing describes the ability of microbes to alter gene regulation according to their local population size. Some successful theory suggests that this is a form of cooperation: investment in shared products is only worthwhile if there are sufficient bacteria making the same product. This theory can explain the genetic diversity in these signaling systems in Gram-positive bacteria such as Bacillus and Staphylococcus. The possible advantages gained by rare genotypes (which can exploit the products of their more common neighbours) could explain why different genotypes can coexist. We show that while these social interactions can occur in simple laboratory experiments they do not occur in naturalistic infections using an invertabrate pathogen, Bacillus thuringiensis. Instead our results suggest that different genotypes are adapted to different-sized hosts. Overall, social models are not easily applied to this system implying that a new explanation for this form of quorum sensing is required.

microbiology↗

Antarctica's wilderness has declined to the exclusion of biodiversity

Recent assessments of the biodiversity value of Earth's dwindling wilderness areas have emphasized the whole of Antarctica as a crucial wilderness in need of urgent protection. Whole-of-continent designations for Antarctic conservation remain controversial, however, because of widespread human impacts and frequently used provisions in Antarctic law for the designation of specially protected areas to conserve wilderness values, species and ecosystems. Here we investigate the extent to which Antarctica's wilderness encompasses its biodiversity. We assembled a comprehensive record of human activity on the continent (~ 2.7 million localities) and used it to identify unvisited areas [≥] 10 000 km2 (i.e. Antarctica's wilderness areas) and their representation of biodiversity. We show that, at best, 7 770 000 km2 of wilderness remains, covering 56.9% of the continent's surface area, however it captures few important biodiversity features. Important Bird Areas, ice-free Antarctic Conservation Biogeographic Regions crucial for biodiversity and areas with verified biodiversity records are largely excluded. Our results demonstrate that Antarcticas wilderness has already declined to the exclusion of much of its biodiversity. But that on a continent set aside as a natural reserve, increased regulation of human activity and urgent expansion of the Antarctic specially protected area network could feasibly reverse this trend.

ecology↗

Targeting antibiotic resistant bacteria with phages reduces bacterial density in an insect host

Phage therapy is attracting growing interest among clinicians as antibiotic resistance continues becoming harder to control. However, clinical trials and animal model studies on bacteriophage treatment are still scarce and results on the efficacy vary. Recent research suggests that using traditional antimicrobials in concert with phage could have desirable synergistic effects that hinder the evolution of resistance. Here, we present a novel insect gut model to study phage-antibiotic interaction in a system where antibiotic resistance initially exists in very low frequency and phage specifically targets the resistance bearing cells. We demonstrate that while phage therapy could not reduce the frequency of target bacteria in the population during positive selection by antibiotics, it alleviated the antibiotic induced blooming by lowering the overall load of resistant cells. The highly structured gut environment had pharmacokinetic effects on both phage and antibiotic dynamics compared to in vitro: antibiotics did not reduce the overall amount of bacteria, demonstrating a simple turnover of gut flora from non-resistant to resistant population with little cost. The results imply moderate potential for using phage as an aid to target antibiotic resistant gut infections, and question the usefulness of in vitro inferences.

evolutionary biology↗

Crystal Toxins and the volunteer’s dilemma in bacteria

1The growth and virulence of the bacteria Bacillus thuringiensis depends on the production of Cry toxins, which are used to perforate the gut of its host. Successful invasion of the host relies on producing a threshold amount of toxin, after which there is no benefit from producing more toxin. Consequently, the production of Cry toxin appears to be a different type of social problem compared with the public goods scenarios that bacteria often encounter. We show that selection for toxin production is a volunteers dilemma. We make the specific predictions that: (1) selection for toxin production depends upon an interplay between the number of bacterial cells that each host ingests, and the genetic relatedness between those cells; (2) cheats that do not produce toxin gain an advantage when at low frequencies, and at high bacterial density, allowing them to be maintained in a population alongside toxin producing cells. More generally, our results emphasise the diversity of the social games that bacteria play.

evolutionary biology↗

Plasmid transfer is biased towards close kin in bacteria from natural populations.

Plasmids play a key role in microbial ecology and evolution, yet the determinants of plasmid transfer rates are poorly understood. Here we investigate the importance of genetic similarity between naturally co-occurring Escherichia coli isolates in the transfer of two plasmids (narrow-host-range R1 and broad-host-range RP4). We uncovered extensive variability, spanning over five orders of magnitude, in the ability of isolates to donate and receive plasmids. Overall, transfer was strongly biased towards clone-mates, but not correlated to genetic distance between donors and recipients. Transfer was limited by the presence of a functional restriction-modification system in recipients, thus bias towards kin might be explained by sharing of identical restriction systems. Such conjugation within lineages sets the stage for longer-term pair-wise coevolutionary interactions between plasmids and bacterial hosts.

ecology↗