PPARα-targeted mitochondrial bioenergetics mediate repair of intestinal barriers at the host-microbe intersection during SIV infection.

Chronic gut inflammatory diseases are associated with disruption of intestinal epithelial barriers and impaired mucosal immunity. HIV-1 (HIV) causes depletion of mucosal CD4+ T cells early in infection and disruption of gut epithelium, resulting in chronic inflammation and immunodeficiency. Although antiretroviral therapy (ART) is effective in suppressing viral replication, it is incapable of restoring the "leaky gut," which poses an impediment for HIV cure efforts. Strategies are needed for rapid repair of the epithelium to protect intestinal microenvironments and immunity in inflamed gut. Using an in vivo nonhuman primate intestinal loop model of HIV/AIDS, we identified the pathogenic mechanism underlying sustained disruption of gut epithelium and explored rapid repair of gut epithelium at the intersection of microbial metabolism. Molecular, immunological, and metabolomic analyses revealed marked loss of peroxisomal proliferator-activated receptor-α (PPARα) signaling, predominant impairment of mitochondrial function, and epithelial disruption both in vivo and in vitro. To elucidate pathways regulating intestinal epithelial integrity, we introduced probiotic Lactobacillus plantarum into Simian immunodeficiency virus (SIV)-inflamed intestinal lumen. Rapid recovery of the epithelium occurred within 5 h of L. plantarum administration, independent of mucosal CD4+ T cell recovery, and in the absence of ART. This intestinal barrier repair was driven by L. plantarum-induced PPARα activation and restoration of mitochondrial structure and fatty acid β-oxidation. Our data highlight the critical role of PPARα at the intersection between microbial metabolism and epithelial repair in virally inflamed gut and as a potential mitochondrial target for restoring gut barriers in other infectious or gut inflammatory diseases

Triclosan Exposure Is Associated with Rapid Restructuring of the Microbiome in Adult Zebrafish

Growing evidence indicates that disrupting the microbial community that comprises the intestinal tract, known as the gut microbiome, can contribute to the development or severity of disease. As a result, it is important to discern the agents responsible for microbiome disruption. While animals are frequently exposed to a diverse array of environmental chemicals, little is known about their effects on gut microbiome stability and structure. Here, we demonstrate how zebrafish can be used to glean insight into the effects of environmental chemical exposure on the structure and ecological dynamics of the gut microbiome. Specifically, we exposed forty-five adult zebrafish to triclosan-laden food for four or seven days or a control diet, and analyzed their microbial communities using 16S rRNA amplicon sequencing. Triclosan exposure was associated with rapid shifts in microbiome structure and diversity. We find evidence that several operational taxonomic units (OTUs) associated with the family Enterobacteriaceae appear to be susceptible to triclosan exposure, while OTUs associated with the genus Pseudomonas appeared to be more resilient and resistant to exposure. We also found that triclosan exposure is associated with topological alterations to microbial interaction networks and results in an overall increase in the number of negative interactions per microbe in these networks. Together these data indicate that triclosan exposure results in altered composition and ecological dynamics of microbial communities in the gut. Our work demonstrates that because zebrafish afford rapid and inexpensive interrogation of a large number of individuals, it is a useful experimental system for the discovery of the gut microbiome’s interaction with environmental chemicals.Data Availability Statement: All sequencing data files are available from the SRA database (accession number PRJNA313944)

Microbiome Variation in an Intertidal Sea Anemone Across Latitudes and Symbiotic States

Many cnidarians form symbiotic relationships with brown dinoflagellate algae in the genus Symbiodinium. Bacteria are important to this symbiosis, with diverse functions such as providing nutrients to the symbiont and pathogen protection to the cnidarian. Disrupted bacterial communities are associated with thermally stressed cnidarians, which have a higher likelihood of expelling their symbionts, an event called bleaching. To better understand the association between thermal tolerance and bacterial community structure, we studied communities associated with an exceptionally thermal tolerant cnidarian, Anthopleura elegantissima. This intertidal symbiotic sea anemone is distributed from the subtropical waters of Baja California to subarctic Alaska, and experiences daily temperature fluctuations of up to 20°C. It is also flexible in its symbioses, predominantly hosting Symbiodinium, but occasionally hosting the green algae Elliptochloris marina or existing without symbionts in an aposymbiotic state. We used 16S rRNA gene amplicon sequencing to characterize the natural variation of microbial communities associated with Anthopleura elegantissima in these three symbiotic states and across a latitudinal gradient. In this study, we identified a core microbiome, made up predominantly of low-abundance taxa. We found that the communities associated with A. elegantissima were weakly linked to latitude. Diversity analyses revealed significantly higher species richness values for microbial communities associated with anemones hosting E. marina. Lastly the microbiome communities associated with different symbiotic states were compositionally distinct. Taken together, our results suggest that the structure of microbial communities associated with these temperate cnidarians is tightly linked to symbiotic state and weakly linked to other biogeographic phenomena

Complex interactive effects of water mold, herbicide, and the fungus Batrachochytrium dendrobatidis on Pacific treefrog Hyliola regilla hosts

Infectious diseases pose a serious threat to global biodiversity. However, their ecological impacts are not independent of environmental conditions. For example, the pathogenic fungus Batrachochytrium dendrobatidis (Bd), which has contributed to population declines and extinctions in many amphibian species, interacts with several environmental factors to influence its hosts, but potential interactions with other pathogens and environmental contaminants are understudied. We examined the combined effects of Bd, a water mold (Achlya sp.), and the herbicide Roundup® Regular (hereafter, Roundup®) on larval Pacific treefrog Hyliola regilla hosts. We employed a 2 wk, fully factorial laboratory experiment with 3 ecologically realistic levels (0, 1, and 2 mg l-1 of active ingredient) of field-formulated Roundup®, 2 Achlya treatments (present and absent), and 2 Bd treatments (present and absent). Our results were consistent with sublethal interactive effects involving all 3 experimental factors. When Roundup® was absent, the proportion of Bd-exposed larvae infected with Bd was elevated in the presence of Achlya, consistent with Achlya acting as a synergistic cofactor that facilitated the establishment of Bd infection. However, this Achlya effect became nonsignificant at 1 mg l-1 of the active ingredient of Roundup® and disappeared at the highest Roundup® concentration. In addition, Roundup® decreased Bd loads among Bd-exposed larvae. Our study suggests complex interactive effects of a water mold and a contaminant on Bd infection in amphibian hosts. Achlya and Roundup® were both correlated with altered patterns of Bd infection, but in different ways, and Roundup® appeared to remove the influence of Achlya on Bd

Effect of Killed PRRSV Vaccine on Gut Microbiota Diversity in Pigs

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically important pathogens affecting the global swine industry. Vaccination is still a main strategy for PRRSV control; however, host factors associated with vaccine efficacy remain poorly understood. Growing evidence suggests that mucosa-associated microbiomes may play a role in the responses to vaccination. In this study, we investigated the effects of a killed virus vaccine on the gut microbiome diversity in pigs. Fecal microbial communities were longitudinally assessed in three groups of pigs (vaccinated/challenged with PRRSV, unvaccinated/challenged with PRRSV, and unvaccinated/unchallenged) before and after vaccination and after viral challenge. We observed significant interaction effects between viral challenge and vaccination on both taxonomic richness and community diversity of the gut microbiota. While some specific taxonomic alterations appear to be enhanced in vaccinated/challenged pigs, others appeared to be more consistent with the levels in control animals (unvaccinated/unchallenged), indicating that vaccination incompletely protects against viral impacts on the microbiome. The abundances of several microbial taxa were further determined to be correlated with the level of viral load and the amount of PRRSV reactive CD4+ and CD8+ T-cells. This study highlights the potential roles of gut microbiota in the response of pigs to vaccination, which may pave the road for the development of novel strategies to enhance vaccine efficacy

Single-cavity dual-modelocked 2.36-µm laser

We present the first dual-modelocked femtosecond oscillator operating beyond 2 µm wavelength. This new class of laser is based on a Cr:ZnS gain medium, an InGaSb SESAM for modelocking, and a two-surface reflective device for spatial duplexing of the two modelocked pulse trains (combs). The laser operates at 2.36 µm, and for each comb, we have achieved a FWHM spectral bandwidth of 30 nm, an average power of over 200 mW, and a pulse duration close to 200 fs. The nominal repetition rate is 242 MHz with a sufficiently large repetition rate difference of 4.17 kHz. We also found that the laser is able to produce stable modelocked pulses over a wide range of output power. This result represents a significant step towards realizing dual-comb applications directly above 2 µm using a single free-running laser.ISSN:1094-408

Aggregative Soil Sampling Using Boot Covers Compared to Soil Grabs From Commercial Romaine Fields Shows Similar Indicator Organism and Microbial Community Recoveries

Aggregative boot cover sampling may be a more representative, practical, and powerful method for preharvest produce soil testing than grab sampling because boot covers aggregate soil from larger areas. Our study tests if boot cover sampling results reflect quality and safety indicator organisms and community diversity of grab sampling. We collected soil samples from commercial romaine lettuce fields spanning 5060 m2 using boot covers (n = 28, m = 1.1 ± 0.4 g; wearing boot covers and walking along the path), composite grabs (n = 28, m = 231 ± 24 g; consisting of 60 grabs of 3–5 g each), and high-resolution grabs (n = 72, m = 56 ± 4 g; taking one sample per stratum). Means and standard deviations of log-transformed aerobic plate counts (APCs) were 7.0 ± 0.3, 7.1 ± 0.2, and 7.3 ± 0.2 log(CFU/g) for boot covers, composite grabs, and high-resolution grabs, respectively. APCs did not show biologically meaningful differences between sample types. Boot covers recovered on average 0.6 log(CFU/g) more total coliforms than both grabs (p < 0.001) where means and standard deviations of log-transformed counts were 3.2 ± 1.0, 2.6 ± 0.6, and 2.6 ± 1.0 log(CFU/g) for boot covers, composite grabs, and high-resolution grabs, respectively. There were no generic E. coli detected in any sample by enumeration methods with LODs of 1.3–2.1 log(CFU/g) for boot covers and 0.5 log(CFU/g) for both grabs. By 16S rRNA sequencing, community species diversity (alpha diversity) was not significantly different within collection methods. While communities differed (p < 0.001) between soil sampling methods (beta diversity), variance in microbial communities was not significantly different. Of the 28 phyla and 297 genera detected, 25 phyla (89%) and 258 genera (87%) were found by all methods. Overall, aggregative boot cover sampling is similar to both grab methods for recovering quality and safety indicator organisms and representative microbiomes. This justifies future work testing aggregative soil sampling for foodborne pathogen detection

High-power low-noise 2-GHz femtosecond laser oscillator at 2.4 µm

Femtosecond lasers with high repetition rates are attractive for spectroscopic applications with high sampling rates, high power per comb line, and resolvable lines. However, at long wavelengths beyond 2 µm, current laser sources are either limited to low output power or repetition rates below 1 GHz. Here we present an ultrafast laser oscillator operating with high output power at multi-GHz repetition rate. The laser produces transform-limited 155-fs pulses at a repetition rate of 2 GHz, and an average power of 0.8 W, reaching up to 0.7 mW per comb line at the center wavelength of 2.38 µm. We have achieved this milestone via a Cr2+-doped ZnS solid-state laser modelocked with an InGaSb/GaSb SESAM. The laser is stable over several hours of operation. The integrated relative intensity noise is 0.15% rms for [10 Hz, 100 MHz], and the laser becomes shot noise limited (-160 dBc/Hz) at frequencies above 10 MHz. Our timing jitter measurements reveal contributions from pump laser noise and relaxation oscillations, with a timing jitter of 100 fs integrated over [3 kHz, 100 MHz]. These results open up a path towards fast and sensitive spectroscopy directly above 2 µm.ISSN:1094-408

Marginal Zinc Deficiency and Environmentally Relevant Concentrations of Arsenic Elicit Combined Effects on the Gut Microbiome

Xenobiotic compounds, such as arsenic, have the potential to alter the composition and functioning of the gut microbiome. The gut microbiome may also interact with these compounds to mediate their impact on the host. However, little is known about how dietary variation may reshape how the microbiome responds to xenobiotic exposures or how these modified responses may in turn impact host physiology. Here, we investigated the combinatorial effects of marginal zinc deficiency and physiologically relevant concentrations of arsenic on the microbiome. Both zinc deficiency and arsenic exposure were individually associated with altered microbial diversity and when combined elicited synergistic effects. Microbial abundance also covaried with host physiological changes, indicating that the microbiome may contribute to or be influenced by these pathologies. Collectively, this work demonstrates that dietary zinc intake influences the sensitivity of the microbiome to subsequent arsenic exposure.Extensive research shows that dietary variation and toxicant exposure impact the gut microbiome, yielding effects on host physiology. However, prior work has mostly considered such exposure-microbiome interactions through the lens of single-factor exposures. In practice, humans exposed to toxicants vary in their dietary nutritional status, and this variation may impact subsequent exposure of the gut microbiome. For example, chronic arsenic exposure affects 200 million people globally and is often comorbid with zinc deficiency. Zinc deficiency can enhance arsenic toxicity, but it remains unknown how zinc status impacts the gut microbiome’s response to arsenic exposure and whether this response links to host toxicity. Using 16S amplicon sequencing, we examined the combinatorial effects of exposure to environmentally relevant concentrations of arsenic on the composition of the microbiome in C57BL/6 mice fed diets varying in zinc concentration. Arsenic exposure and marginal zinc deficiency independently altered microbiome diversity. When combined, their effects on microbiome community structure were amplified. Generalized linear models identified microbial taxa whose relative abundance in the gut was perturbed by zinc deficiency, arsenic, or their interaction. Further, we correlated taxonomic abundances with host DNA damage, adiponectin expression, and plasma zinc concentration to identify taxa that may mediate host physiological responses to arsenic exposure or zinc deficiency. Arsenic exposure and zinc restriction also result in increased DNA damage and decreased plasma zinc. These physiological changes are associated with the relative abundance of several gut taxa. These data indicate that marginal zinc deficiency sensitizes the microbiome to arsenic exposure and that the microbiome associates with some toxicological effects of arsenic