David Fast

Comparative evaluation of the genomes of three common Drosophila-associated bacteria.

ABSTRACT Drosophila melanogaster is an excellent model to explore the molecular exchanges that occur between an animal intestine and associated microbes. Previous studies in Drosophila uncovered a sophisticated web of host responses to intestinal bacteria. The outcomes of these responses define critical events in the host, such as the establishment of immune responses, access to nutrients, and the rate of larval development. Despite our steady march towards illuminating the host machinery that responds to bacterial presence in the gut, there are significant gaps in our understanding of the microbial products that influence bacterial association with a fly host. We sequenced and characterized the genomes of three common Drosophila-associated microbes: Lactobacillus plantarum, Lactobacillus brevis and Acetobacter pasteurianus. For each species, we compared the genomes of Drosophila-associated strains to the genomes of strains isolated from alternative sources. We found that environmental Lactobacillus strains readily associated with adult Drosophila and were similar to fly isolates in terms of genome organization. In contrast, we identified a strain of A. pasteurianus that apparently fails to associate with adult Drosophila due to an inability to grow on fly nutrient food. Comparisons between association competent and incompetent A. pasteurianus strains identified a short list of candidate genes that may contribute to survival on fly medium. Many of the gene products unique to fly-associated strains have established roles in the stabilization of host-microbe interactions. These data add to a growing body of literature that examines the microbial perspective of host-microbe relationships. Summary: We examined the genomes of Drosophila-associated bacteria to identify factors that allow survival within the host. These preliminary studies may point at bacterial products that influence host health.

Mono-Association with Lactobacillus plantarum Disrupts Intestinal Homeostasis in adult Drosophila.

The microbiome of Drosophila promotes intestinal stem cell division through evolutionarily conserved biochemical pathways. As such, axenic flies have lower rates of gut stem cell division than age-matched wild type counterparts. Additionally, flies with a full consortium of symbiotic bacteria are shorter lived than those maintained in the absence of a microbiome. However, we do not know if stem cell division is essential for symbiont-dependent regulation of adult fly lifespan. To determine if individual symbionts cause aging-dependent death in Drosophila, we examined the impacts of common symbionts on host longevity. In this study, we found that mono-association of adult Drosophila with Lactobacillus plantarum, a widely reported fly symbiont, and member of the probiotic Lactobacillus genus, curtails adult longevity relative to germ-free counterparts. However, the effects of plantarum on lifespan were independent of intestinal aging. Instead, we found that association with plantarum causes an extensive intestinal pathology within the host, characterized by loss of intestinal stem cells, impaired epithelial renewal, and a gradual erosion of epithelial integrity. Our study uncovers an unknown aspect of Lactobacillus plantarum-Drosophila interactions, and establishes a simple model to characterize symbiont-dependent disruption of intestinal homeostasis.

Genomewide association analyses of fitness traits in captive-reared Chinook salmon: Applications in evaluating conservation strategies

A novel application of genomewide association analyses is to use trait‐associated loci to monitor the effects of conservation strategies on potentially adaptive genetic variation. Comparisons of fitness between captive‐ and wild‐origin individuals, for example, do not reveal how captive rearing affects genetic variation underlying fitness traits or which traits are most susceptible to domestication selection. Here, we used data collected across four generations to identify loci associated with six traits in adult Chinook salmon (Oncorhynchus tshawytscha) and then determined how two alternative management approaches for captive rearing affected variation at these loci. Loci associated with date of return to freshwater spawning grounds (return timing), length and weight at return, age at maturity, spawn timing, and daily growth coefficient were identified using 9108 restriction site‐associated markers and random forest, an approach suitable for polygenic traits. Mapping of trait‐associated loci, gene annotations, and integration of results across multiple studies revealed candidate regions involved in several fitness‐related traits. Genotypes at trait‐associated loci were then compared between two hatchery populations that were derived from the same source but are now managed as separate lines, one integrated with and one segregated from the wild population. While no broad‐scale change was detected across four generations, there were numerous regions where trait‐associated loci overlapped with signatures of adaptive divergence previously identified in the two lines. Many regions, primarily with loci linked to return and spawn timing, were either unique to or more divergent in the segregated line, suggesting that these traits may be responding to domestication selection. This study is one of the first to utilize genomic approaches to demonstrate the effectiveness of a conservation strategy, managed gene flow, on trait‐associated—and potentially adaptive—loci. The results will promote the development of trait‐specific tools to better monitor genetic change in captive and wild populations.

Commensal pathogen competition impacts host viability

Significance Enteric pathogens including the causative agent of cholera, Vibrio cholerae, use the type-six secretion system (T6SS) to kill commensal microbes in the host intestine. Eradicating competing microbes allows pathogens to improve colonization. However, it is not known whether commensal destruction has additional consequences on host viability. We used the Drosophila model of cholera to determine the impacts of T6SS on fly health and longevity. We found that T6SS-dependent competition with the symbiotic Acetobacter pasteurianus intensified disease symptoms, and accelerated host death. Gnotobiotic flies without A. pasteurianus abolished T6SS-dependent death, and reintroduction of A. pasteurianus alone was sufficient to restore accelerated death. These observations implicate T6SS-dependent interactions with commensal bacteria as a factor for the progression of cholera. While the structure and regulatory networks that govern type-six secretion system (T6SS) activity of Vibrio cholerae are becoming increasingly clear, we know less about the role of T6SS in disease. Under laboratory conditions, V. cholerae uses T6SS to outcompete many Gram-negative species, including other V. cholerae strains and human commensal bacteria. However, the role of these interactions has not been resolved in an in vivo setting. We used the Drosophila melanogaster model of cholera to define the contribution of T6SS to V. cholerae pathogenesis. Here, we demonstrate that interactions between T6SS and host commensals impact pathogenesis. Inactivation of T6SS, or removal of commensal bacteria, attenuates disease severity. Reintroduction of the commensal, Acetobacter pasteurianus, into a germ-free host is sufficient to restore T6SS-dependent pathogenesis in which T6SS and host immune responses regulate viability. Together, our data demonstrate that T6SS acts on commensal bacteria to promote the pathogenesis of V. cholerae.

Monoassociation with Lactobacillus plantarum Disrupts Intestinal Homeostasis in Adult Drosophila melanogaster

ABSTRACT Adult Drosophila melanogaster raised in the absence of symbiotic bacteria have fewer intestinal stem cell divisions and a longer life span than their conventionally reared counterparts. However, we do not know if increased stem cell divisions are essential for symbiont-dependent regulation of longevity. To determine if individual symbionts cause aging-dependent death in Drosophila, we examined the impacts of common symbionts on host longevity. We found that monoassociation of adult Drosophila with Lactobacillus plantarum, a widely reported fly symbiont and member of the probiotic Lactobacillus genus, curtails adult longevity relative to germfree counterparts. The effects of Lactobacillus plantarum on life span were independent of intestinal aging. Instead, we found that association with Lactobacillus plantarum causes an extensive intestinal pathology within the host, characterized by loss of stem cells, impaired epithelial renewal, and a gradual erosion of epithelial ultrastructure. Our study uncovers an unknown aspect of Lactobacillus plantarum-Drosophila interactions and establishes a simple model to characterize symbiont-dependent disruption of intestinal homeostasis. IMPORTANCE Under homeostatic conditions, gut bacteria provide molecular signals that support the organization and function of the host intestine. Sudden shifts in the composition or distribution of gut bacterial communities impact host receipt of bacterial cues and disrupt tightly regulated homeostatic networks. We used the Drosophila melanogaster model to determine the effects of prominent fly symbionts on host longevity and intestinal homeostasis. We found that monoassociation with Lactobacillus plantarum leads to a loss of intestinal progenitor cells, impaired epithelial renewal, and disruption of gut architecture as flies age. These observations uncover a novel phenotype caused by monoassociation of a germfree host with a common symbiont and establish a simple model to characterize symbiont-dependent loss of intestinal homeostasis. IMPORTANCE Under homeostatic conditions, gut bacteria provide molecular signals that support the organization and function of the host intestine. Sudden shifts in the composition or distribution of gut bacterial communities impact host receipt of bacterial cues and disrupt tightly regulated homeostatic networks. We used the Drosophila melanogaster model to determine the effects of prominent fly symbionts on host longevity and intestinal homeostasis. We found that monoassociation with Lactobacillus plantarum leads to a loss of intestinal progenitor cells, impaired epithelial renewal, and disruption of gut architecture as flies age. These observations uncover a novel phenotype caused by monoassociation of a germfree host with a common symbiont and establish a simple model to characterize symbiont-dependent loss of intestinal homeostasis.

The symbiont Lactobacillus plantarum causes intestinal pathogenesis in adult Drosophila.

The microbiome of Drosophila promotes intestinal stem cell division through evolutionarily conserved biochemical pathways. As such, axenic flies have lower rates of gut stem cell division than age-matched wild type counterparts. Additionally, flies with a full consortium of symbiotic bacteria are shorter lived than those maintained in the absence of a microbiome. However, we do not know if stem cell division is essential for symbiont-dependent regulation of adult fly lifespan. To determine if individual symbionts cause aging-dependent death in Drosophila, we examined the impacts of common symbionts on host longevity. In this study, we found that mono-association of adult Drosophila with Lactobacillus plantarum, a widely reported fly symbiont, and member of the probiotic Lactobacillus genus, curtails adult longevity relative to germ-free counterparts. However, the effects of plantarum on lifespan were independent of intestinal aging. Instead, we found that association with plantarum causes an extensive intestinal pathology within the host, characterized by loss of intestinal stem cells, impaired epithelial renewal, and a gradual erosion of epithelial integrity. Our study uncovers an unknown aspect of Lactobacillus plantarum-Drosophila interactions, and establishes a simple model to characterize symbiont-dependent disruption of intestinal homeostasis.

Lactobacillus plantarum is a pathobiont for adult Drosophila.

The microbiome of Drosophila promotes intestinal stem cell division through evolutionarily conserved biochemical pathways. As such, axenic flies have lower rates of gut stem cell division than age-matched wild type counterparts. Additionally, flies with a full consortium of symbiotic bacteria are shorter lived than those maintained in the absence of a microbiome. However, we do not know if stem cell division is essential for symbiont-dependent regulation of adult fly lifespan. To determine if individual symbionts cause aging-dependent death in Drosophila, we examined the impacts of common symbionts on host longevity. In this study, we found that mono-association of adult Drosophila with Lactobacillus plantarum, a widely reported fly symbiont, and member of the probiotic Lactobacillus genus, curtails adult longevity relative to germ-free counterparts. However, the effects of plantarum on lifespan were independent of intestinal aging. Instead, we found that association with plantarum causes an extensive intestinal pathology within the host, characterized by loss of intestinal stem cells, impaired epithelial renewal, and a gradual erosion of epithelial integrity. Our study uncovers an unknown aspect of Lactobacillus plantarum-Drosophila interactions, and establishes a simple model to characterize symbiont-dependent disruption of intestinal homeostasis.

What can genomics tell us about the success of enhancement programs in anadromous Chinook salmon? A comparative analysis across four generations

Population enhancement through the release of cultured organisms can be an important tool for marine restoration. However, there has been considerable debate about whether releases effectively contribute to conservation and harvest objectives, and whether cultured organisms impact the fitness of wild populations. Pacific salmonid hatcheries on the West Coast of North America represent one of the largest enhancement programs in the world. Molecular-based pedigree studies on one or two generations have contributed to our understanding of the fitness of hatchery-reared individuals relative to wild individuals, and tend to show that hatchery fish have lower reproductive success. However, interpreting the significance of these results can be challenging because the long-term genetic and ecological effects of releases on supplemented populations are unknown. Further, pedigree studies have been opportunistic, rather than hypothesis driven, and have not provided information on “best case” management scenarios. Here, we present a comparative, experimental approach based on genome-wide surveys of changes in diversity in two hatchery lines founded from the same population. We demonstrate that gene flow with wild individuals can reduce divergence from the wild source population over four generations. We also report evidence for consistent genetic changes in a closed hatchery population that can be explained by both genetic drift and domestication selection. The results of this study suggest that genetic risks can be minimized over at least four generations with appropriate actions, and provide empirical support for a decision-making framework that is relevant to the management of hatchery populations.

Comparative evaluation of the genomes of common bacterial members of the Drosophila intestinal community

Drosophila melanogaster is an excellent model to explore the molecular exchanges that occur between an animal intestine and their microbial passengers. For example, groundbreaking studies in flies uncovered a sophisticated web of host responses to intestinal bacteria. The outcomes of these responses define critical events in the host, such as the establishment of immune responses, access to nutrients, and the rate of larval development. Despite our steady march towards illuminating the host machinery that responds to bacterial presence in the gut, we know remarkably little about the microbial products that influence bacterial association with a fly host. To address this deficiency, we sequenced and characterized the genomes of three common Drosophila-associated microbes: Lactobacillus plantarum, Lactobacillus brevis and Acetobacter pasteurianus. In each case, we compared the genomes of Drosophila-associated strains to the genomes of strains isolated from alternative sources. This approach allowed us to identify molecular functions common to Drosophila-associated microbes, and, in the case of A. pasteurianus, to identify genes that are essential for association with the host. Of note, many of the gene products unique to fly-associated strains have established roles in the stabilization of host-microbe interactions. We believe that these data provide a valuable starting point for a more thorough examination of the microbial perspective on host-microbe relationships.