Increased Dietary Trp, Thr, and Met Supplementation Improves Performance, Health, and Protein Metabolism of Weaned Piglets under Mixed Management and Poor Housing Conditions.

A sanitary challenge was carried out to induce suboptimal herd health while investigating the effect of amino acids supplementation on piglet responses. Weaned piglets of high sanitary status (6.33 ± 0.91 kg of BW) were distributed in a 2 × 2 factorial arrangement into two similar facilities with contrasting sanitary conditions and two different diets. Our results suggest that increased Trp, Thr, and Met dietary supplementation could support the immune systems of piglets under a sanitary challenge. In this manner, AA+ supplementation improved the performance and metabolism of piglets under mixed management and poor sanitary conditions. No major temporal microbiome changes were associated with differences in performance regardless of sanitary conditions or diets. Since piglets often become mixed in multiple-site production systems and facility hygiene is also often neglected, this study suggests that increased Trp, Thr, and Met (AA+) dietary supplementation could contribute to mitigating the side effects of these harmful risk factors in modern pig farms.

Antibiotic-induced gut dysbiosis elicits gut-brain axis relevant multi-omic signatures and behavioral and neuroendocrine changes in a nonhuman primate model.

Emerging evidence indicates that antibiotic-induced dysbiosis can play an etiological role in the pathogenesis of neuropsychiatric disorders. However, most of this evidence comes from rodent models. The objective of this study was to evaluate if antibiotic-induced gut dysbiosis can elicit changes in gut metabolites and behavior indicative of gut-brain axis disruption in common marmosets (Callithrix jacchus) - a nonhuman primate model often used to study sociability and stress. We were able to successfully induce dysbiosis in marmosets using a custom antibiotic cocktail (vancomycin, enrofloxacin and neomycin) administered orally for 28 days. This gut dysbiosis altered gut metabolite profiles, behavior, and stress reactivity. Increase in gut Fusobacterium spp. post-antibiotic administration was a novel dysbiotic response and has not been observed in any rodent or human studies to date. There were significant changes in concentrations of several gut metabolites which are either neurotransmitters (e.g., GABA and serotonin) or have been found to be moderators of gut-brain axis communication in rodent models (e.g., short-chain fatty acids and bile acids). There was an increase in affiliative behavior and sociability in antibiotic-administered marmosets, which might be a coping mechanism in response to gut dysbiosis-induced stress. Increase in urinary cortisol levels after multiple stressors provides more definitive proof that this model of dysbiosis may cause disrupted communication between gut and brain in common marmosets. This study is a first attempt to establish common marmosets as a novel model to study the impact of severe gut dysbiosis on gut-brain axis cross-talk and behavior.

Salmonella enterica induces biogeography-specific changes in the gut microbiome of pigs.

Swine are a major reservoir of an array of zoonotic Salmonella enterica subsp. enterica lineage I serovars including Derby, Typhimurium, and 4,[5],12:i:- (a.k.a. Monophasic Typhimurium). In this study, we assessed the gastrointestinal (GI) microbiome composition of pigs in different intestinal compartments and the feces following infection with specific zoonotic serovars of S. enterica (S. Derby, S. Monophasic, and S. Typhimurium). 16S rRNA based microbiome analysis was performed to assess for GI microbiome changes in terms of diversity (alpha and beta), community structure and volatility, and specific taxa alterations across GI biogeography (small and large intestine, feces) and days post-infection (DPI) 2, 4, and 28; these results were compared to disease phenotypes measured as histopathological changes. As previously reported, only S. Monophasic and S. Typhimurium induced morphological alterations that marked an inflammatory milieu restricted to the large intestine in this experimental model. S. Typhimurium alone induced significant changes at the alpha- (Simpson's and Shannon's indexes) and beta-diversity levels, specifically at the peak of inflammation in the large intestine and feces. Increased community dispersion and volatility in colonic apex and fecal microbiomes were also noted for S. Typhimurium. All three Salmonella serovars altered community structure as measured by co-occurrence networks; this was most prominent at DPI 2 and 4 in colonic apex samples. At the genus taxonomic level, a diverse array of putative short-chain fatty acid (SCFA) producing bacteria were altered and often decreased during the peak of inflammation at DPI 2 and 4 within colonic apex and fecal samples. Among all putative SCFA producing bacteria, Prevotella showed a broad pattern of negative correlation with disease scores at the peak of inflammation. In addition, Prevotella 9 was found to be significantly reduced in all Salmonella infected groups compared to the control at DPI 4 in the colonic apex. In conclusion, this work further elucidates that distinct swine-related zoonotic serovars of S. enterica can induce both shared (high resilience) and unique (altered resistance) alterations in gut microbiome biogeography, which helps inform future investigations of dietary modifications aimed at increasing colonization resistance against Salmonella through GI microbiome alterations.

Pushing the limits of detectability: mixed dark matter from strong gravitational lenses.

One of the frontiers for advancing what is known about dark matter lies in using strong gravitational lenses to characterize the population of the smallest dark matter haloes. There is a large volume of information in strong gravitational lens images - the question we seek to answer is to what extent we can refine this information. To this end, we forecast the detectability of a mixed warm and cold dark matter scenario using the anomalous flux ratio method from strong gravitational lensed images. The halo mass function of the mixed dark matter scenario is suppressed relative to cold dark matter but still predicts numerous low-mass dark matter haloes relative to warm dark matter. Since the strong lensing signal receives a contribution from a range of dark matter halo masses and since the signal is sensitive to the specific configuration of dark matter haloes, not just the halo mass function, degeneracies between different forms of suppression in the halo mass function, relative to cold dark matter, can arise. We find that, with a set of lenses with different configurations of the main deflector and hence different sensitivities to different mass ranges of the halo mass function, the different forms of suppression of the halo mass function between the warm dark matter model and the mixed dark matter model can be distinguished with 40 lenses with Bayesian odds of 30:1.

The waxy mutation in sorghum and other cereal grains reshapes the gut microbiome by reducing levels of multiple beneficial species.

Waxy starches from cereal grains contain >90% amylopectin due to naturally occurring mutations that block amylose biosynthesis. Waxy starches have unique organoleptic characteristics (e.g. sticky rice) as well as desirable physicochemical properties for food processing. Using isogenic pairs of wild type sorghum lines and their waxy derivatives, we studied the effects of waxy starches in the whole grain context on the human gut microbiome. In vitro fermentations with human stool microbiomes show that beneficial taxonomic and metabolic signatures driven by grain from wild type parental lines are lost in fermentations of grain from the waxy derivatives and the beneficial signatures can be restored by addition of resistant starch. These undesirable effects are conserved in fermentations of waxy maize, wheat, rice and millet. We also demonstrate that humanized gnotobiotic mice fed low fiber diets supplemented with 20% grain from isogenic pairs of waxy vs. wild type parental sorghum have significant differences in microbiome composition and show increased weight gain. We conclude that the benefits of waxy starches on food functionality can have unintended tradeoff effects on the gut microbiome and host physiology that could be particularly relevant in human populations consuming large amounts of waxy grains.

Evidence for a Causal Role for Escherichia coli Strains Identified as Adherent-Invasive (AIEC) in Intestinal Inflammation.

Enrichment of adherent-invasive Escherichia coli (AIEC) has been consistently detected in subsets of inflammatory bowel disease (IBD) patients. Although some AIEC strains cause colitis in animal models, these studies did not systematically compare AIEC with non-AIEC strains, and causal links between AIEC and disease are still disputed. Specifically, it remains unclear whether AIEC shows enhanced pathogenicity compared to that of commensal E. coli found in the same ecological microhabitat and if the in vitro phenotypes used to classify strains as AIEC are pathologically relevant. Here, we utilized in vitro phenotyping and a murine model of intestinal inflammation to systematically compare strains identified as AIEC with those identified as non-AIEC and relate AIEC phenotypes to pathogenicity. Strains identified as AIEC caused, on average, more severe intestinal inflammation. Intracellular survival/replication phenotypes routinely used to classify AIEC positively correlated with disease, while adherence to epithelial cells and tumor necrosis factor alpha production by macrophages did not. This knowledge was then applied to design and test a strategy to prevent inflammation by selecting E. coli strains that adhered to epithelial cells but poorly survived/replicated intracellularly. Two E. coli strains that ameliorated AIEC-mediated disease were subsequently identified. In summary, our results show a relationship between intracellular survival/replication in E. coli and pathology in murine colitis, suggesting that strains possessing these phenotypes might not only become enriched in human IBD but also contribute to disease. We provide new evidence that specific AIEC phenotypes are pathologically relevant and proof of principle that such mechanistic information can be therapeutically exploited to alleviate intestinal inflammation. IMPORTANCE Inflammatory bowel disease (IBD) is associated with an altered gut microbiota composition, including expansion of Proteobacteria. Many species in this phylum are thought to contribute to disease under certain conditions, including adherent-invasive Escherichia coli (AIEC) strains, which are enriched in some patients. However, whether this bloom contributes to disease or is just a response to IBD-associated physiological changes is unknown. Although assigning causality is challenging, appropriate animal models can test the hypothesis that AIEC strains have an enhanced ability to cause colitis in comparison to other gut commensal E. coli strains and to identify bacterial traits contributing to virulence. We observed that AIEC strains are generally more pathogenic than commensal E. coli and that bacterial intracellular survival/replication phenotypes contributed to disease. We also found that E. coli strains lacking primary virulence traits can prevent inflammation. Our findings provide critical information on E. coli pathogenicity that may inform development of IBD diagnostic tools and therapies.

The effect of dietary coenzyme Q10 supplementation on egg quality and liver histopathology of layer quails under cadmium challenge.

The demand for quail eggs has been increased over the last decade due to its beneficial nutritional quality characteristics; however, different nutritional and environmental stressors adversely impact the quality of the produced eggs. This study was conducted to investigate whether dietary supplementation of coenzyme Q10 (CoQ10) could mitigate the negative impact of cadmium (Cd) administration on egg quality and liver histopathology. A total of 162 six-week-old laying Japanese quail (Coturnix japonica) were randomly allotted into three experimental groups. Treatments were as follows: (1) negative control (NC): feeding basal diet; (2) positive control (PC): feeding basal diet and Cd administration; and (3) CdQ10: feeding basal diet supplemented with CoQ10 (900 mg/kg diet) and Cd administration. Cadmium (10 mg/kg BW) was subcutaneously administrated at 10 and 11 weeks of age (woa). Feed conversion ratio (FCR), egg production, egg mass, mortality rate, Cd residue in egg, liver histopathology, and some internal and external egg quality indices were evaluated. Administration of Cd increased FCR in the PC group, but supplementation of CoQ10 partially ameliorated the impact of Cd on FCR (p < 0.05). Cadmium administration decreased both egg production and egg mass; however, CoQ10 supplementation partially mitigated these adverse effects of Cd injection in the CdQ10 compared to the PC group (p < 0.05). Cadmium decreased eggshell thickness and Haugh unit in PC quail compared to both NC and CdQ10 quail (p < 0.05). Moreover, egg yolk colour intensity was enhanced by CoQ10, where a* and b* indices were higher in CdQ10 compared to PC (p < 0.05). In conclusion, the current results demonstrate the beneficial effects of dietary CoQ10 supplementation on liver histopathology and some egg quality indices of Cd-challenged quail.

Constraints on Sterile Neutrino Models from Strong Gravitational Lensing, Milky Way Satellites, and the Lyman-α Forest.

The nature of dark matter is one of the most important unsolved questions in science. Some darkf matter candidates do not have sufficient nongravitational interactions to be probed in laboratory or accelerator experiments. It is thus important to develop astrophysical probes which can constrain or lead to a discovery of such candidates. We illustrate this using state-of-the-art measurements of strong gravitationally lensed quasars to constrain four of the most popular sterile neutrino models, and also report the constraints for other independent methods that are comparable in procedure. First, we derive effective relations to describe the correspondence between the mass of a thermal relic warm dark matter particle and the mass of sterile neutrinos produced via Higgs decay and grand unified theory (GUT)-scale scenarios, in terms of large-scale structure and galaxy formation astrophysical effects. Second, we show that sterile neutrinos produced through the Higgs decay mechanism are allowed only for mass >26 keV, and GUT-scale scenario >5.3 keV. Third, we show that the single sterile neutrino model produced through active neutrino oscillations is allowed for mass >92 keV, and the three sterile neutrino minimal standard model (νMSM) for mass >16 keV. These are the most stringent experimental limits on these models.

Genetic analysis of seed traits in Sorghum bicolor that affect the human gut microbiome.

Prebiotic fibers, polyphenols and other molecular components of food crops significantly affect the composition and function of the human gut microbiome and human health. The abundance of these, frequently uncharacterized, microbiome-active components vary within individual crop species. Here, we employ high throughput in vitro fermentations of pre-digested grain using a human microbiome to identify segregating genetic loci in a food crop, sorghum, that alter the composition and function of human gut microbes. Evaluating grain produced by 294 sorghum recombinant inbreds identifies 10 loci in the sorghum genome associated with variation in the abundance of microbial taxa and/or microbial metabolites. Two loci co-localize with sorghum genes regulating the biosynthesis of condensed tannins. We validate that condensed tannins stimulate the growth of microbes associated with these two loci. Our work illustrates the potential for genetic analysis to systematically discover and characterize molecular components of food crops that influence the human gut microbiome.

The Unique Seed Protein Composition of Quality Protein Popcorn Promotes Growth of Beneficial Bacteria From the Human Gut Microbiome.

The effects of fiber, complex carbohydrates, lipids, and small molecules from food matrices on the human gut microbiome have been increasingly studied. Much less is known about how dietary protein can influence the composition and function of the gut microbial community. Here, we used near-isogenic maize lines of conventional popcorn and quality-protein popcorn (QPP) to study the effects of the opaque-2 mutation and associated quality-protein modifiers on the human gut microbiome. Opaque-2 blocks the synthesis of major maize seed proteins (α-zeins), resulting in a compensatory synthesis of new seed proteins that are nutritionally beneficial with substantially higher levels of the essential amino acids lysine and tryptophan. We show that QPP lines stimulate greater amounts of butyrate production by human gut microbiomes in in vitro fermentation of popped and digested corn from parental and QPP hybrids. In human gut microbiomes derived from diverse individuals, bacterial taxa belonging to the butyrate-producing family Lachnospiraceae, including the genera Coprococcus and Roseburia were consistently increased when fermenting QPP vs. parental popcorn lines. We conducted molecular complementation to further demonstrate that lysine-enriched seed protein can stimulate growth and butyrate production by microbes through distinct pathways. Our data show that organisms such as Coprococcus can utilize lysine and that other gut microbes, such as Roseburia spp., instead, utilize fructoselysine produced during thermal processing (popping) of popcorn. Thus, the combination of seed composition in QPP and interaction of protein adducts with carbohydrates during thermal processing can stimulate the growth of health-promoting, butyrate-producing organisms in the human gut microbiome through multiple pathways.

Quantitative Protein Analysis of ZPB2, ZPB1 and ZPC in the Germinal Disc and a Non-Germinal Disc Region of the Inner Perivitelline Layer in Two Genetic Lines of Turkey Hens That Differ in Fertility.

The avian inner perivitelline layer (IPVL), containing the zona pellucida (ZP) family of proteins, surrounds the ovulated ovum. In mammalian species, ZP proteins serve as key component(s) in binding sperm and initiating the acrosome reaction. Sperm binding at the germinal disc (GD) region of the IPVL initiates fertilization in avian species, and the amount of sperm binding at the GD reflects female fertility. The current research determined whether reported differences in mRNA expression in two genetic lines of turkey hens (E, high fertility and F, low fertility) translated to the protein level. ZPB2 in the IPVL is greater in the GD region compared with the nongerminal disc (NGD) region, as indicated by both mRNA and protein expression. However, protein expressions of ZPB1 and ZPC in the IPVL of E- and F-line turkey hens was in contrast to previously reported mRNA expression. The results indicate that the mRNA expression of ZP proteins at their site of synthesis in E- and F-line hens often does not directly correlate with the IPVL abundance of these proteins. The greater protein concentration of ZPB2 in the GD region compared with the NGD regions suggests that this protein may be critical for sperm binding at the GD region.

Systems-Based Approach for Optimization of Assembly-Free Bacterial MLST Mapping.

Epidemiological surveillance of bacterial pathogens requires real-time data analysis with a fast turnaround, while aiming at generating two main outcomes: (1) species-level identification and (2) variant mapping at different levels of genotypic resolution for population-based tracking and surveillance, in addition to predicting traits such as antimicrobial resistance (AMR). Multi-locus sequence typing (MLST) aids this process by identifying sequence types (ST) based on seven ubiquitous genome-scattered loci. In this paper, we selected one assembly-dependent and one assembly-free method for ST mapping and applied them with the default settings and ST schemes they are distributed with, and systematically assessed their accuracy and scalability across a wide array of phylogenetically divergent Public Health-relevant bacterial pathogens with available MLST databases. Our data show that the optimal k-mer length for stringMLST is species-specific and that genome-intrinsic and -extrinsic features can affect the performance and accuracy of the program. Although suitable parameters could be identified for most organisms, there were instances where this program may not be directly deployable in its current format. Next, we integrated stringMLST into our freely available and scalable hierarchical-based population genomics platform, ProkEvo, and further demonstrated how the implementation facilitates automated, reproducible bacterial population analysis.

Pain management consultation for acute pancreatitis: impact on length of stay and opioid utilization.

Aim: The purpose of the study was to understand the impact of a pain management consult for acute pancreatitis patients on their inpatient length of stay, morphine milligram equivalences (MMEs) and pancreatitis severity. Materials & methods: Adult patient data were extracted from the electronic health records from 1 October 2016 to 31 December 2018. Results & conclusion: Of 277 patients with a single acute pancreatitis hospitalization, 23 had a pain consultation (treatment group), whereas 254 did not (control group). There were statistically significant differences in median length of stay, median MME total and median MME per day between the treatment and control groups with comparable severity and pain scores (6.8 vs 3.1 days, 196.5 vs 33.8 MMEs, 30.9 vs 12.1 MMEs, respectively, p < 0.0001). This study emphasizes the complexity of pain management and the importance of further research in the field.

Heuristic Mining of Hierarchical Genotypes and Accessory Genome Loci in Bacterial Populations.

Routine and systematic use of bacterial whole-genome sequencing (WGS) is enhancing the accuracy and resolution of epidemiological investigations carried out by Public Health laboratories and regulatory agencies. Large volumes of publicly available WGS data can be used to study pathogenic populations at a large scale. Recently, a freely available computational platform called ProkEvo was published to enable reproducible, automated, and scalable hierarchical-based population genomic analyses using bacterial WGS data. This implementation of ProkEvo demonstrated the importance of combining standard genotypic mapping of populations with mining of accessory genomic content for ecological inference. In particular, the work highlighted here used ProkEvo-derived outputs for population-scaled hierarchical analyses using the R programming language. The main objective was to provide a practical guide for microbiologists, ecologists, and epidemiologists by showing how to: i) use a phylogeny-guided mapping of hierarchical genotypes; ii) assess frequency distributions of genotypes as a proxy for ecological fitness; iii) determine kinship relationships and genetic diversity using specific genotypic classifications; and iv) map lineage differentiating accessory loci. To enhance reproducibility and portability, R markdown files were used to demonstrate the entire analytical approach. The example dataset contained genomic data from 2,365 isolates of the zoonotic foodborne pathogen Salmonella Newport. Phylogeny-anchored mapping of hierarchical genotypes (Serovar -> BAPS1 -> ST -> cgMLST) revealed the population genetic structure, highlighting sequence types (STs) as the keystone differentiating genotype. Across the three most dominant lineages, ST5 and ST118 shared a common ancestor more recently than with the highly clonal ST45 phylotype. ST-based differences were further highlighted by the distribution of accessory antimicrobial resistance (AMR) loci. Lastly, a phylogeny-anchored visualization was used to combine hierarchical genotypes and AMR content to reveal the kinship structure and lineage-specific genomic signatures. Combined, this analytical approach provides some guidelines for conducting heuristic bacterial population genomic analyses using pan-genomic information.

Reproductive toxicity of roundup®-treated feed on broiler breeder roosters and the amelioration of these deleterious effects with inclusion of humic acids in feed.

Glyphosate-based herbicides (GBHs) such as RoundUp® are a staple of modern crop production, and as a result, residues of their ingredients are typically found in animal feeds. GBH ingredients have repeatedly been shown to impact the male reproductive health of various animals, but at present, the impact of GBH exposures on reproductive health have not been investigated in broiler breeder roosters. This study sought to determine the effect of these exposures on roosters as well as the ability of humic acids (HA) to counteract the effect of GBH exposure. Through 18 weeks of treatment with Roundup®-added or HA-added feeds compared against a common broiler breeder mash, negative effects of Roundup® exposure were seen on testis morphology as well as sperm quality. Increased exposure to Roundup® ingredients resulted in increased vacuolation of seminiferous tubule epithelium. Exposure to Roundup® impacted assessments of sperm quality including sperm mobility, viability and count during the experimental trail. HA supplementation served as a promising adsorptive additive by improving both morphology and sperm quality during the trial. Roundup® exposure was shown to have a negative influence on broiler breeder rooster reproductive health while HA improved reproductive health. The use of HA as an absorbent additive in broiler breeder feeds shows promise in improving reproductive efficiency in broiler breeders.

Effects of coenzyme Q10 on reproductive performance of laying Japanese quail (Coturnix japonica) under cadmium challenge.

Japanese quail is an increasingly important bird of economic importance for commercial egg and meat production, particularly in developing countries. There is a need for research aimed at improving efficiency of these birds during stressful challenges, such as oxidative stress. Coenzyme Q10 (CoQ10), a highly functional antioxidant, protects cells against oxidative stress. This study was conducted to determine the effects of CoQ10 on reproductive performance of Japanese quail under cadmium (Cd) challenge. A total of 216 six-wk-old Japanese quail were randomly allocated into 3 groups for an 8 wk experimental trial. The treatments include a negative control (NC): feeding basal diet; a positive control (PC): feeding basal diet and cadmium administration (1 mg/100 g BW, at 10 and 11 wk of age), and (CdQ10): feeding CoQ10 supplemented (900 mg/kg diet) basal diet and Cd administration. At 11 and 13 wk of age, egg production, body weight, mortality, oviduct, and ovarian biometry, were recorded. Histology and histopathology of isthmus and magnum, fertility, hatchability, hatchling quality, and HSP70 mRNA transcript abundance in the utero-vaginal junction (UVJ) were evaluated. Positive control and CdQ10 group had no significant effect on live body weight, stroma weight, follicle size, hatchability, and fertility; however, Cd administration increased (P < 0.01) mortality rate in the PC group compared to the NC and CdQ10 groups. CdQ10 quail produced more eggs and had a higher hatchling quality compared to the PC group (P < 0.01). The thickness and height of isthmus and magnum folds in the CdQ10 group was increased compared to the PC group (P < 0.01) and overall oviduct weight was increased with CoQ10 supplementation (P < 0.01). Compared to PC, the CdQ10 group had a reduction in infiltration of inflammatory cells. Relative abundance of HSP70 mRNA in UVJ was influenced by interactive effect of treatment × time (P < 0.05). In conclusion, dietary supplementation of CoQ10 showed beneficial effects on some reproduction characteristics of female Japanese quail under Cd-induced oxidative stress.

Captive Common Marmosets (Callithrix jacchus) Are Colonized throughout Their Lives by a Community of Bifidobacterium Species with Species-Specific Genomic Content That Can Support Adaptation to Distinct Metabolic Niches.

The common marmoset (Callithrix jacchus) is an omnivorous New World primate whose diet in the wild includes large amounts of fruit, seeds, flowers, and a variety of lizards and invertebrates. Marmosets also feed heavily on tree gums and exudates, and they have evolved unique morphological and anatomical characteristics to facilitate gum feeding (gummivory). In this study, we characterized the fecal microbiomes of adult and infant animals from a captive population of common marmosets at the Callitrichid Research Center at the University of Nebraska at Omaha under their normal dietary and environmental conditions. The microbiomes of adult animals were dominated by species of Bifidobacterium, Bacteroides, Prevotella, Phascolarctobacterium, Megamonas, and Megasphaera. Culturing and genomic analysis of the Bifidobacterium populations from adult animals identified four known marmoset-associated species (B. reuteri, B. aesculapii, B. myosotis, and B. hapali) and three unclassified taxa of Bifidobacterium that are phylogenetically distinct. Species-specific quantitative PCR (qPCR) confirmed that these same species of Bifidobacterium are abundant members of the microbiome throughout the lives of the animals. Genomic loci in each Bifidobacterium species encode enzymes to support growth and major marmoset milk oligosaccharides during breastfeeding; however, metabolic islands that can support growth on complex polysaccharide substrates in the diets of captive adults (pectin, xyloglucan, and xylan), including loci in B. aesculapii that can support its unique ability to grow on arabinogalactan-rich tree gums, were species-specific. IMPORTANCEBifidobacterium species are recognized as important, beneficial microbes in the human gut microbiome, and their ability colonize individuals at different stages of life is influenced by host, dietary, environmental, and ecological factors, which is poorly understood. The common marmoset is an emerging nonhuman primate model with a short maturation period, making this model amenable to study the microbiome throughout a life history. Features of the microbiome in captive marmosets are also shared with human gut microbiomes, including abundant populations of Bifidobacterium species. Our studies show that several species of Bifidobacterium are dominant members of the captive marmoset microbiome throughout their life history. Metabolic capacities in genomes of the marmoset Bifidobacterium species suggest species-specific adaptations to different components of the captive marmoset diet, including the unique capacity in B. aesculapii for degradation of gum arabic, suggesting that regular dietary exposure in captivity may be important for preserving gum-degrading species in the microbiome.

ProkEvo: an automated, reproducible, and scalable framework for high-throughput bacterial population genomics analyses.

Whole Genome Sequence (WGS) data from bacterial species is used for a variety of applications ranging from basic microbiological research, diagnostics, and epidemiological surveillance. The availability of WGS data from hundreds of thousands of individual isolates of individual microbial species poses a tremendous opportunity for discovery and hypothesis-generating research into ecology and evolution of these microorganisms. Flexibility, scalability, and user-friendliness of existing pipelines for population-scale inquiry, however, limit applications of systematic, population-scale approaches. Here, we present ProkEvo, an automated, scalable, reproducible, and open-source framework for bacterial population genomics analyses using WGS data. ProkEvo was specifically developed to achieve the following goals: (1) Automation and scaling of complex combinations of computational analyses for many thousands of bacterial genomes from inputs of raw Illumina paired-end sequence reads; (2) Use of workflow management systems (WMS) such as Pegasus WMS to ensure reproducibility, scalability, modularity, fault-tolerance, and robust file management throughout the process; (3) Use of high-performance and high-throughput computational platforms; (4) Generation of hierarchical-based population structure analysis based on combinations of multi-locus and Bayesian statistical approaches for classification for ecological and epidemiological inquiries; (5) Association of antimicrobial resistance (AMR) genes, putative virulence factors, and plasmids from curated databases with the hierarchically-related genotypic classifications; and (6) Production of pan-genome annotations and data compilation that can be utilized for downstream analysis such as identification of population-specific genomic signatures. The scalability of ProkEvo was measured with two datasets comprising significantly different numbers of input genomes (one with ~2,400 genomes, and the second with ~23,000 genomes). Depending on the dataset and the computational platform used, the running time of ProkEvo varied from ~3-26 days. ProkEvo can be used with virtually any bacterial species, and the Pegasus WMS uniquely facilitates addition or removal of programs from the workflow or modification of options within them. To demonstrate versatility of the ProkEvo platform, we performed a hierarchical-based population structure analyses from available genomes of three distinct pathogenic bacterial species as individual case studies. The specific case studies illustrate how hierarchical analyses of population structures, genotype frequencies, and distribution of specific gene functions can be integrated into an analysis. Collectively, our study shows that ProkEvo presents a practical viable option for scalable, automated analyses of bacterial populations with direct applications for basic microbiology research, clinical microbiological diagnostics, and epidemiological surveillance.