Multistate Outbreak Investigation of Salmonella Infections Linked to Kratom: A Focus on Traceback, Laboratory, and Regulatory Activities.

ABSTRACT: During spring 2018, the U.S. Food and Drug Administration (FDA), Centers for Disease Control and Prevention, and state and local public health agencies responded to a multistate outbreak of gastrointestinal illnesses caused by multiple Salmonella serovars and associated with consumption of kratom, a product harvested from a tropical tree native to Southeast Asia. The outbreak included 199 case-patients reported by 41 U.S. states, with illness onset dates ranging 11 from January 2017 to 8 May 2018, leading to 54 hospitalizations and no deaths. Case-patients reported purchasing kratom products from physical and online retail points of service (POSs). Products distributed to 16 POSs where 24 case-patients from 17 states purchased kratom were selected for traceback investigation. Traceback revealed that the kratom was imported from several countries, the most common being Indonesia. Local and state officials collected product samples from case-patients and retail POSs. The FDA collected 76 product samples from POSs and distributors, of which 42 (55%) tested positive for Salmonella. The positive samples exhibited a range of pulsed-field gel electrophoresis patterns and whole genome sequence genetic heterogeneity, and 25 (60%) of 42 samples yielded at least one isolate indistinguishable from one or more outbreak-related clinical isolates. Although it does not exclude a possibility of a single contamination source, the extent of genetic diversity exhibited by the Salmonella isolates recovered from product samples and a lack of traceback convergence suggested that kratom was widely contaminated across multiple sites from which it was grown, harvested, and packaged. As a result of the contamination, kratom products were recalled by numerous firms (both voluntarily and mandatory). Epidemiologic, traceback, and laboratory evidence supported the conclusion that kratom products were associated with illnesses.

Call for a framework for reporting evidence for life beyond Earth.

Our generation could realistically be the one to discover evidence of life beyond Earth. With this privileged potential comes responsibility. The magnitude of the question of whether we are alone in the Universe, and the public interest therein, opens the possibility that results may be taken to imply more than the observations support, or than the observers intend. As life-detection objectives become increasingly prominent in space sciences, it is essential to open a community dialogue about how to convey information in a subject matter that is diverse, complicated and has a high potential to be sensationalized. Establishing best practices for communicating about life detection can serve to set reasonable expectations on the early stages of a hugely challenging endeavour, attach value to incremental steps along the path, and build public trust by making clear that false starts and dead ends are an expected and potentially productive part of the scientific process. Here we endeavour to motivate and seed the discussion with basic considerations and offer an example of how such considerations might be incorporated and applied in a proof-of-concept-level framework. Everything mentioned herein, including the name of the confidence scale, is intended not as a prescription, but simply as the beginning of an important dialogue.

Exoplanet Biosignatures: Observational Prospects.

Exoplanet hunting efforts have revealed the prevalence of exotic worlds with diverse properties, including Earth-sized bodies, which has fueled our endeavor to search for life beyond the Solar System. Accumulating experiences in astrophysical, chemical, and climatological characterization of uninhabitable planets are paving the way to characterization of potentially habitable planets. In this paper, we review our possibilities and limitations in characterizing temperate terrestrial planets with future observational capabilities through the 2030s and beyond, as a basis of a broad range of discussions on how to advance "astrobiology" with exoplanets. We discuss the observability of not only the proposed biosignature candidates themselves but also of more general planetary properties that provide circumstantial evidence, since the evaluation of any biosignature candidate relies on its context. Characterization of temperate Earth-sized planets in the coming years will focus on those around nearby late-type stars. The James Webb Space Telescope (JWST) and later 30-meter-class ground-based telescopes will empower their chemical investigations. Spectroscopic studies of potentially habitable planets around solar-type stars will likely require a designated spacecraft mission for direct imaging, leveraging technologies that are already being developed and tested as part of the Wide Field InfraRed Survey Telescope (WFIRST) mission. Successful initial characterization of a few nearby targets will be an important touchstone toward a more detailed scrutiny and a larger survey that are envisioned beyond 2030. The broad outlook this paper presents may help develop new observational techniques to detect relevant features as well as frameworks to diagnose planets based on the observables. Key Words: Exoplanets-Biosignatures-Characterization-Planetary atmospheres-Planetary surfaces. Astrobiology 18, 739-778.

Exoplanet Biosignatures: Future Directions.

We introduce a Bayesian method for guiding future directions for detection of life on exoplanets. We describe empirical and theoretical work necessary to place constraints on the relevant likelihoods, including those emerging from better understanding stellar environment, planetary climate and geophysics, geochemical cycling, the universalities of physics and chemistry, the contingencies of evolutionary history, the properties of life as an emergent complex system, and the mechanisms driving the emergence of life. We provide examples for how the Bayesian formalism could guide future search strategies, including determining observations to prioritize or deciding between targeted searches or larger lower resolution surveys to generate ensemble statistics and address how a Bayesian methodology could constrain the prior probability of life with or without a positive detection. Key Words: Exoplanets-Biosignatures-Life detection-Bayesian analysis. Astrobiology 18, 779-824.

Exoplanet Biosignatures: At the Dawn of a New Era of Planetary Observations.

The rapid rate of discoveries of exoplanets has expanded the scope of the science possible for the remote detection of life beyond Earth. The Exoplanet Biosignatures Workshop Without Walls (EBWWW) held in 2016 engaged the international scientific community across diverse scientific disciplines, to assess the state of the science and technology in the search for life on exoplanets, and to identify paths for progress. The workshop activities resulted in five major review papers, which provide (1) an encyclopedic review of known and proposed biosignatures and models used to ascertain them (Schwieterman et al., 2018 in this issue); (2) an in-depth review of O2 as a biosignature, rigorously examining the nuances of false positives and false negatives for evidence of life (Meadows et al., 2018 in this issue); (3) a Bayesian framework to comprehensively organize current understanding to quantify confidence in biosignature assessments (Catling et al., 2018 in this issue); (4) an extension of that Bayesian framework in anticipation of increasing planetary data and novel concepts of biosignatures (Walker et al., 2018 in this issue); and (5) a review of the upcoming telescope capabilities to characterize exoplanets and their environment (Fujii et al., 2018 in this issue). Because of the immense content of these review papers, this summary provides a guide to their complementary scope and highlights salient features. Strong themes that emerged from the workshop were that biosignatures must be interpreted in the context of their environment, and that frameworks must be developed to link diverse forms of scientific understanding of that context to quantify the likelihood that a biosignature has been observed. Models are needed to explore the parameter space where measurements will be widespread but sparse in detail. Given the technological prospects for large ground-based telescopes and space-based observatories, the detection of atmospheric signatures of a few potentially habitable planets may come before 2030. Key Words: Exoplanets-Biosignatures-Remote observation-Spectral imaging-Bayesian analysis. Astrobiology 18, 619-626.

Exoplanet Biosignatures: Understanding Oxygen as a Biosignature in the Context of Its Environment.

We describe how environmental context can help determine whether oxygen (O2) detected in extrasolar planetary observations is more likely to have a biological source. Here we provide an in-depth, interdisciplinary example of O2 biosignature identification and observation, which serves as the prototype for the development of a general framework for biosignature assessment. Photosynthetically generated O2 is a potentially strong biosignature, and at high abundance, it was originally thought to be an unambiguous indicator for life. However, as a biosignature, O2 faces two major challenges: (1) it was only present at high abundance for a relatively short period of Earth's history and (2) we now know of several potential planetary mechanisms that can generate abundant O2 without life being present. Consequently, our ability to interpret both the presence and absence of O2 in an exoplanetary spectrum relies on understanding the environmental context. Here we examine the coevolution of life with the early Earth's environment to identify how the interplay of sources and sinks may have suppressed O2 release into the atmosphere for several billion years, producing a false negative for biologically generated O2. These studies suggest that planetary characteristics that may enhance false negatives should be considered when selecting targets for biosignature searches. We review the most recent knowledge of false positives for O2, planetary processes that may generate abundant atmospheric O2 without a biosphere. We provide examples of how future photometric, spectroscopic, and time-dependent observations of O2 and other aspects of the planetary environment can be used to rule out false positives and thereby increase our confidence that any observed O2 is indeed a biosignature. These insights will guide and inform the development of future exoplanet characterization missions. Key Words: Biosignatures-Oxygenic photosynthesis-Exoplanets-Planetary atmospheres. Astrobiology 18, 630-662.

Exoplanet Biosignatures: A Framework for Their Assessment.

Finding life on exoplanets from telescopic observations is an ultimate goal of exoplanet science. Life produces gases and other substances, such as pigments, which can have distinct spectral or photometric signatures. Whether or not life is found with future data must be expressed with probabilities, requiring a framework of biosignature assessment. We present a framework in which we advocate using biogeochemical "Exo-Earth System" models to simulate potential biosignatures in spectra or photometry. Given actual observations, simulations are used to find the Bayesian likelihoods of those data occurring for scenarios with and without life. The latter includes "false positives" wherein abiotic sources mimic biosignatures. Prior knowledge of factors influencing planetary inhabitation, including previous observations, is combined with the likelihoods to give the Bayesian posterior probability of life existing on a given exoplanet. Four components of observation and analysis are necessary. (1) Characterization of stellar (e.g., age and spectrum) and exoplanetary system properties, including "external" exoplanet parameters (e.g., mass and radius), to determine an exoplanet's suitability for life. (2) Characterization of "internal" exoplanet parameters (e.g., climate) to evaluate habitability. (3) Assessment of potential biosignatures within the environmental context (components 1-2), including corroborating evidence. (4) Exclusion of false positives. We propose that resulting posterior Bayesian probabilities of life's existence map to five confidence levels, ranging from "very likely" (90-100%) to "very unlikely" (<10%) inhabited. Key Words: Bayesian statistics-Biosignatures-Drake equation-Exoplanets-Habitability-Planetary science. Astrobiology 18, 709-738.

The Habitability of Proxima Centauri b: Environmental States and Observational Discriminants.

Proxima Centauri b provides an unprecedented opportunity to understand the evolution and nature of terrestrial planets orbiting M dwarfs. Although Proxima Cen b orbits within its star's habitable zone, multiple plausible evolutionary paths could have generated different environments that may or may not be habitable. Here, we use 1-D coupled climate-photochemical models to generate self-consistent atmospheres for several evolutionary scenarios, including high-O2, high-CO2, and more Earth-like atmospheres, with both oxic and anoxic compositions. We show that these modeled environments can be habitable or uninhabitable at Proxima Cen b's position in the habitable zone. We use radiative transfer models to generate synthetic spectra and thermal phase curves for these simulated environments, and use instrument models to explore our ability to discriminate between possible planetary states. These results are applicable not only to Proxima Cen b but to other terrestrial planets orbiting M dwarfs. Thermal phase curves may provide the first constraint on the existence of an atmosphere. We find that James Webb Space Telescope (JWST) observations longward of 10 µm could characterize atmospheric heat transport and molecular composition. Detection of ocean glint is unlikely with JWST but may be within the reach of larger-aperture telescopes. Direct imaging spectra may detect O4 absorption, which is diagnostic of massive water loss and O2 retention, rather than a photosynthetic biosphere. Similarly, strong CO2 and CO bands at wavelengths shortward of 2.5 µm would indicate a CO2-dominated atmosphere. If the planet is habitable and volatile-rich, direct imaging will be the best means of detecting habitability. Earth-like planets with microbial biospheres may be identified by the presence of CH4-which has a longer atmospheric lifetime under Proxima Centauri's incident UV-and either photosynthetically produced O2 or a hydrocarbon haze layer. Key Words: Planetary habitability and biosignatures-Planetary atmospheres-Exoplanets-Spectroscopic biosignatures-Planetary science-Proxima Centauri b. Astrobiology 18, 133-189.

Organic Haze as a Biosignature in Anoxic Earth-like Atmospheres.

Early Earth may have hosted a biologically mediated global organic haze during the Archean eon (3.8-2.5 billion years ago). This haze would have significantly impacted multiple aspects of our planet, including its potential for habitability and its spectral appearance. Here, we model worlds with Archean-like levels of carbon dioxide orbiting the ancient Sun and an M4V dwarf (GJ 876) and show that organic haze formation requires methane fluxes consistent with estimated Earth-like biological production rates. On planets with high fluxes of biogenic organic sulfur gases (CS2, OCS, CH3SH, and CH3SCH3), photochemistry involving these gases can drive haze formation at lower CH4/CO2 ratios than methane photochemistry alone. For a planet orbiting the Sun, at 30× the modern organic sulfur gas flux, haze forms at a CH4/CO2 ratio 20% lower than at 1× the modern organic sulfur flux. For a planet orbiting the M4V star, the impact of organic sulfur gases is more pronounced: at 1× the modern Earth organic sulfur flux, a substantial haze forms at CH4/CO2 ∼ 0.2, but at 30× the organic sulfur flux, the CH4/CO2 ratio needed to form haze decreases by a full order of magnitude. Detection of haze at an anomalously low CH4/CO2 ratio could suggest the influence of these biogenic sulfur gases and therefore imply biological activity on an exoplanet. When these organic sulfur gases are not readily detectable in the spectrum of an Earth-like exoplanet, the thick organic haze they can help produce creates a very strong absorption feature at UV-blue wavelengths detectable in reflected light at a spectral resolution as low as 10. In direct imaging, constraining CH4 and CO2 concentrations will require higher spectral resolution, and R > 170 is needed to accurately resolve the structure of the CO2 feature at 1.57 µm, likely the most accessible CO2 feature on an Archean-like exoplanet. Key Words: Organic haze-Organic sulfur gases-Biosignatures-Archean Earth. Astrobiology 18, 311-329.

Carbon Monoxide and the Potential for Prebiotic Chemistry on Habitable Planets around Main Sequence M Stars.

Lifeless planets with CO2 atmospheres produce CO by CO2 photolysis. On planets around M dwarfs, CO is a long-lived atmospheric compound, as long as UV emission due to the star's chromospheric activity lasts, and the sink of CO and O2 in seawater is small compared to its atmospheric production. Atmospheres containing reduced compounds, like CO, may undergo further energetic and chemical processing to give rise to organic compounds of potential importance for the origin of life. We calculated the yield of organic compounds from CO2-rich atmospheres of planets orbiting M dwarf stars, which were previously simulated by Domagal-Goldman et al. (2014) and Harman et al. (2015), by cosmic rays and lightning using results of experiments by Miyakawa et al. (2002) and Schlesinger and Miller ( 1983a , 1983b ). Stellar protons from active stars may be important energy sources for abiotic synthesis and increase production rates of biological compounds by at least 2 orders of magnitude compared to cosmic rays. Simple compounds such as HCN and H2CO are more readily synthesized than more complex ones, such as amino acids and uracil (considered here as an example), resulting in higher yields for the former and lower yields for the latter. Electric discharges are most efficient when a reducing atmosphere is present. Nonetheless, atmospheres with high quantities of CO2 are capable of producing higher amounts of prebiotic compounds, given that CO is constantly produced in the atmosphere. Our results further support planetary systems around M dwarf stars as candidates for supporting life or its origin. Key Words: Prebiotic chemistry-M dwarfs-Habitable planets-Cosmic rays-Lightning-Stellar activity. Astrobiology 16, 744-754.

The Pale Orange Dot: The Spectrum and Habitability of Hazy Archean Earth.

Recognizing whether a planet can support life is a primary goal of future exoplanet spectral characterization missions, but past research on habitability assessment has largely ignored the vastly different conditions that have existed in our planet's long habitable history. This study presents simulations of a habitable yet dramatically different phase of Earth's history, when the atmosphere contained a Titan-like, organic-rich haze. Prior work has claimed a haze-rich Archean Earth (3.8-2.5 billion years ago) would be frozen due to the haze's cooling effects. However, no previous studies have self-consistently taken into account climate, photochemistry, and fractal hazes. Here, we demonstrate using coupled climate-photochemical-microphysical simulations that hazes can cool the planet's surface by about 20 K, but habitable conditions with liquid surface water could be maintained with a relatively thick haze layer (τ ∼ 5 at 200 nm) even with the fainter young Sun. We find that optically thicker hazes are self-limiting due to their self-shielding properties, preventing catastrophic cooling of the planet. Hazes may even enhance planetary habitability through UV shielding, reducing surface UV flux by about 97% compared to a haze-free planet and potentially allowing survival of land-based organisms 2.7-2.6 billion years ago. The broad UV absorption signature produced by this haze may be visible across interstellar distances, allowing characterization of similar hazy exoplanets. The haze in Archean Earth's atmosphere was strongly dependent on biologically produced methane, and we propose that hydrocarbon haze may be a novel type of spectral biosignature on planets with substantial levels of CO2. Hazy Archean Earth is the most alien world for which we have geochemical constraints on environmental conditions, providing a useful analogue for similar habitable, anoxic exoplanets. Key Words: Haze-Archean Earth-Exoplanets-Spectra-Biosignatures-Planetary habitability. Astrobiology 16, 873-899.

IDENTIFYING PLANETARY BIOSIGNATURE IMPOSTORS: SPECTRAL FEATURES OF CO AND O4 RESULTING FROM ABIOTIC O2/O3 PRODUCTION.

O2 and O3 have been long considered the most robust individual biosignature gases in a planetary atmosphere, yet multiple mechanisms that may produce them in the absence of life have been described. However, these abiotic planetary mechanisms modify the environment in potentially identifiable ways. Here we briefly discuss two of the most detectable spectral discriminants for abiotic O2/O3: CO and O4. We produce the first explicit self-consistent simulations of these spectral discriminants as they may be seen by James Webb Space Telescope (JWST). If JWST-NIRISS and/or NIRSpec observe CO (2.35, 4.6 µm) in conjunction with CO2 (1.6, 2.0, 4.3 µm) in the transmission spectrum of a terrestrial planet it could indicate robust CO2 photolysis and suggest that a future detection of O2 or O3 might not be biogenic. Strong O4 bands seen in transmission at 1.06 and 1.27 µm could be diagnostic of a post-runaway O2-dominated atmosphere from massive H-escape. We find that for these false positive scenarios, CO at 2.35 µm, CO2 at 2.0 and 4.3 µm, and O4 at 1.27 µm are all stronger features in transmission than O2/O3 and could be detected with S/Ns ≳ 3 for an Earth-size planet orbiting a nearby M dwarf star with as few as 10 transits, assuming photon-limited noise. O4 bands could also be sought in UV/VIS/NIR reflected light (at 0.345, 0.36, 0.38, 0.445, 0.475, 0.53, 0.57, 0.63, 1.06, and 1.27 µm) by a next generation direct-imaging telescope such as LUVOIR/HDST or HabEx and would indicate an oxygen atmosphere too massive to be biologically produced.

13C and 15N fractionation of CH4/N2 mixtures during photochemical aerosol formation: Relevance to Titan.

The ratios of the stable isotopes that comprise each chemical species in Titan's atmosphere provide critical information towards understanding the processes taking place within its modern and ancient atmosphere. Several stable isotope pairs, including 12C/13C and 14N/15N, have been measured in situ or probed spectroscopically by Cassini-borne instruments, space telescopes, or through ground-based observations. Current attempts to model the observed isotope ratios incorporate fractionation resulting from atmospheric diffusion, hydrodynamic escape, and primary photochemical processes. However, the effect of a potentially critical pathway for isotopic fractionation - organic aerosol formation and subsequent deposition onto the surface of Titan - has not been considered due to insufficient data regarding fractionation during aerosol formation. To better understand the nature of this process, we have conducted a laboratory study to measure the isotopic fractionation associated with the formation of Titan aerosol analogs, commonly referred to as 'tholins', via far-UV irradiation of several methane (CH4) and dinitrogen (N2) mixtures. Analysis of the δ13C and δ15N isotopic signatures of the photochemical aerosol products using an isotope ratio mass spectrometer (IRMS) show that fractionation direction and magnitude are dependent on the initial bulk composition of the gas mixture. In general, the aerosols showed enrichment in 13C and 14N, and the observed fractionation trends can provide insight into the chemical mechanisms controlling photochemical aerosol formation.

Using biogenic sulfur gases as remotely detectable biosignatures on anoxic planets.

We used one-dimensional photochemical and radiative transfer models to study the potential of organic sulfur compounds (CS(2), OCS, CH(3)SH, CH(3)SCH(3), and CH(3)S(2)CH(3)) to act as remotely detectable biosignatures in anoxic exoplanetary atmospheres. Concentrations of organic sulfur gases were predicted for various biogenic sulfur fluxes into anoxic atmospheres and were found to increase with decreasing UV fluxes. Dimethyl sulfide (CH(3)SCH(3), or DMS) and dimethyl disulfide (CH(3)S(2)CH(3), or DMDS) concentrations could increase to remotely detectable levels, but only in cases of extremely low UV fluxes, which may occur in the habitable zone of an inactive M dwarf. The most detectable feature of organic sulfur gases is an indirect one that results from an increase in ethane (C(2)H(6)) over that which would be predicted based on the planet's methane (CH(4)) concentration. Thus, a characterization mission could detect these organic sulfur gases-and therefore the life that produces them-if it could sufficiently quantify the ethane and methane in the exoplanet's atmosphere.

A revised, hazy methane greenhouse for the Archean Earth.

Geological and biological evidence suggests that Earth was warm during most of its early history, despite the fainter young Sun. Upper bounds on the atmospheric CO2 concentration in the Late Archean/Paleoproterozoic (2.8-2.2 Ga) from paleosol data suggest that additional greenhouse gases must have been present. Methanogenic bacteria, which were arguably extant at that time, may have contributed to a high concentration of atmospheric CH4, and previous calculations had indicated that a CH4-CO2-H2O greenhouse could have produced warm Late Archean surface temperatures while still satisfying the paleosol constraints on pCO2. Here, we revisit this conclusion. Correction of an error in the CH4 absorption coefficients, combined with the predicted early onset of climatically cooling organic haze, suggest that the amount of greenhouse warming by CH4 was more limited and that pCO2 must therefore have been 0.03 bar, at or above the upper bound of the value obtained from paleosols. Enough warming from CH4 remained in the Archean, however, to explain why Earth's climate cooled and became glacial when atmospheric O2 levels rose in the Paleoproterozoic. Our new model also shows that greenhouse warming by higher hydrocarbon gases, especially ethane (C2H6), may have helped to keep the Late Archean Earth warm.