Synechococcus sp. PCC7335 responses to far-red enriched spectra and anoxic/microoxic atmospheres: Potential for astrobiotechnological applications.

Recently, cyanobacteria have gained attention in space exploration to support long-term crewed missions via Bioregenerative Life Support Systems. In this frame, cyanobacteria would provide biomass and profitable biomolecules through oxygenic photosynthesis, uptaking CO2, and releasing breathable O2. Their growth potential and organic matter production will depend on their ability to photoacclimate to different light intensities and spectra, maximizing incident light harvesting. Studying cyanobacteria responses to different light regimes will also benefit the broader field of astrobiology, providing data on the possibility of oxygenic photosynthetic life on planets orbiting stars with emission spectra different than the Sun. Here, we tested the acclimation and productivity of Synechococcus sp. PCC7335 (hereafter PCC7335), capable of Far-Red Light Photoacclimation (FaRLiP) and type III chromatic acclimation (CA3), in an anoxic, CO2-enriched atmosphere and under a spectrum simulating the low energetic light regime of an M-dwarf star, also comparable to a subsuperficial environment. When exposed to the light spectrum, with few photons in the visible (VIS) and rich in far-red (FR), PCC7335 did not activate FaRLiP but acclimated only via CA3, achieving a biomass productivity higher than expected, considering the low VIS light availability, and a higher production of phycocyanin, a valuable pigment, with respect to solar light. Its growth or physiological responses of PCC7335 were not affected by the anoxic atmosphere. In these conditions, PCC7335 efficiently produced O2 and scavenged CO2. Results highlight the photosynthetic plasticity of PCC7335, its suitability for astrobiotechnological applications, and the importance to investigate biodiversity of oxygenic photosynthesis for searching life beyond Earth.

Raman Spectroscopy Applied to Early Detection of Clostridium Infection in Milk.

Detecting Clostridium in milk presents a significant challenge for the dairy industry given that traditional methods are time-consuming and not specific for these bacteria. Microbiological techniques are expensive and require qualified personnel. Clostridium, in the form of spores, can withstand pasteurization and revert to its vegetative form during cheese aging. These gas-producing bacteria are known for their production of carbon dioxide and hydrogen, causing the formation of slits, cracks, and irregular eyes in hard and semi-hard cheeses. However, gas analysis in the vial headspace of appropriate culture can be exploited to specifically detect Clostridium presence, since the closest competing bacterial Bacilli produces only carbon dioxide. The aim of this paper is to present a Raman-spectroscopy-based instrument for a rapid, inexpensive identification of Clostridium in milk with a limit of detection of 29 spores/L. The proposed measurement procedure is analog to that routinely used, based on the most probable number method. The Raman-based instrument speeds up the detection of a vial's positivity. A test conducted with Clostridium spores demonstrated its effectiveness in almost halving the time needed for the measurement campaign compared to the traditional method.

Static, refractive and monolithic Fourier transform spectrometer: development and prototyping.

Static Fourier transform spectrometers are devices that can be realized as monolithic and compact assemblies. In the "grating-based" monolithic version, they are usually realized gluing together a beam-splitter with two reflective diffraction gratings using spacers as connecting elements. In this work we present the development and test of an alternative form of this kind of instrument in which the dispersive elements are Littrow's prisms and are glued to the splitting element, forming in this way a robust and filled structure with no air gaps. The device can work in the visible/near infrared spectral region with a resolution power that varies across the spectral range due to the dispersion of the used glasses. The absence of hollow regions inside the monolithic block makes the device extremely robust and protects the optical surfaces inside the interferometer from possible contaminations. The device can be easily miniaturized, as it does not require spacers or structural elements other than just the optical parts. The tested instrument works in the 470-850 nm wavelength range with a variable resolution between 3000 and 300.

Growth and Photosynthetic Efficiency of Microalgae and Plants with Different Levels of Complexity Exposed to a Simulated M-Dwarf Starlight.

Oxygenic photosynthetic organisms (OPOs) are primary producers on Earth and generate surface and atmospheric biosignatures, making them ideal targets to search for life from remote on Earth-like exoplanets orbiting stars different from the Sun, such as M-dwarfs. These stars emit very low light in the visible and most light in the far-red, an issue for OPOs, which mostly utilize visible light to photosynthesize and grow. After successfully testing procaryotic OPOs (cyanobacteria) under a simulated M-dwarf star spectrum (M7, 365-850 nm) generated through a custom-made lamp, we tested several eukaryotic OPOs: microalgae (Dixoniella giordanoi, Microchloropsis gaditana, Chromera velia, Chlorella vulgaris), a non-vascular plant (Physcomitrium patens), and a vascular plant (Arabidopsis thaliana). We assessed their growth and photosynthetic efficiency under three light conditions: M7, solar (SOL) simulated spectra, and far-red light (FR). Microalgae grew similarly in SOL and M7, while the moss P. patens showed slower growth in M7 with respect to SOL. A. thaliana grew similarly in SOL and M7, showing traits typical of shade-avoidance syndrome. Overall, the synergistic effect of visible and far-red light, also known as the Emerson enhancing effect, could explain the growth in M7 for all organisms. These results lead to reconsidering the possibility and capability of the growth of OPOs and are promising for finding biosignatures on exoplanets orbiting the habitable zone of distant stars.

Oxygenic photosynthetic responses of cyanobacteria exposed under an M-dwarf starlight simulator: Implications for exoplanet's habitability.

Introduction: The search for life on distant exoplanets is expected to rely on atmospheric biosignatures detection, such as oxygen of biological origin. However, it is not demonstrated how much oxygenic photosynthesis, which on Earth depends on visible light, could work under spectral conditions simulating exoplanets orbiting the Habitable Zone of M-dwarf stars, which have low light emission in the visible and high light emission in the far-red/near-infrared. By utilizing cyanobacteria, the first organisms to evolve oxygenic photosynthesis on our planet, and a starlight simulator capable of accurately reproducing the emission spectrum of an M-dwarf in the range 350-900 nm, we could answer this question. Methods: We performed experiments with the cyanobacterium Chlorogloeopsis fritschii PCC6912, capable of Far-Red Light Photoacclimation (FaRLiP), which allows the strain to harvest far-red in addition to visible light for photosynthesis, and Synechocystis sp. PCC6803, a species unable to perform this photoacclimation, comparing their responses when exposed to three simulated light spectra: M-dwarf, solar and far-red. We analysed growth and photosynthetic acclimation features in terms of pigment composition and photosystems organization. Finally, we determined the oxygen production of the strains directly exposed to the different spectra. Results: Both cyanobacteria were shown to grow and photosynthesize similarly under M-dwarf and solar light conditions: Synechocystis sp. by utilizing the few photons in the visible, C. fritschii by harvesting both visible and far-red light, activating the FaRLiP response. Discussion: Our results experimentally show that an M-dwarf light spectrum could support a biological oxygen production similar to that in solar light at the tested light intensities, suggesting the possibility to discover such atmospheric biosignatures on those exoplanets if other boundary conditions are met.

Transcriptomic and photosynthetic analyses of Synechocystis sp. PCC6803 and Chlorogloeopsis fritschii sp. PCC6912 exposed to an M-dwarf spectrum under an anoxic atmosphere.

Introduction: Cyanobacteria appeared in the anoxic Archean Earth, evolving for the first time oxygenic photosynthesis and deeply changing the atmosphere by introducing oxygen. Starting possibly from UV-protected environments, characterized by low visible and far-red enriched light spectra, cyanobacteria spread everywhere on Earth thanks to their adaptation capabilities in light harvesting. In the last decade, few cyanobacteria species which can acclimate to far-red light through Far-Red Light Photoacclimation (FaRLiP) have been isolated. FaRLiP cyanobacteria were thus proposed as model organisms to study the origin of oxygenic photosynthesis as well as its possible functionality around stars with high far-red emission, the M-dwarfs. These stars are astrobiological targets, as their longevity could sustain life evolution and they demonstrated to host rocky terrestrial-like exoplanets within their Habitable Zone. Methods: We studied the acclimation responses of the FaRLiP strain Chlorogloeopsis fritschii sp. PCC6912 and the non-FaRLiP strain Synechocystis sp. PCC6803 to the combination of three simulated light spectra (M-dwarf, solar and far-red) and two atmospheric compositions (oxic, anoxic). We first checked their growth, O2 production and pigment composition, then we studied their transcriptional responses by RNA sequencing under each combination of light spectrum and atmosphere conditions. Results and discussion: PCC6803 did not show relevant differences in gene expression when comparing the responses to M-dwarf and solar-simulated lights, while far-red caused a variation in the transcriptional level of many genes. PCC6912 showed, on the contrary, different transcriptional responses to each light condition and activated the FaRLiP response under the M-dwarf simulated light. Surprisingly, the anoxic atmosphere did not impact the transcriptional profile of the 2 strains significantly. Results show that both cyanobacteria seem inherently prepared for anoxia and to harvest the photons emitted by a simulated M-dwarf star, whether they are only visible (PCC6803) or also far-red photons (PCC6912). They also show that visible photons in the simulated M-dwarf are sufficient to keep a similar metabolism with respect to solar-simulated light. Conclusion: Results prove the adaptability of the cyanobacterial metabolism and enhance the plausibility of finding oxygenic biospheres on exoplanets orbiting M-dwarf stars.

High sensitivity static Fourier transform spectrometer.

Static Fourier transform spectrometers (S-FTSs) are well-consolidated instruments providing high throughput and high spectral resolution in a narrow spectral band. They use two reflective gratings as dispersive elements in a Michelson interferometer. Gratings allow high spectral dispersion and consequently high resolution, but, due to the light diffused from their grooves, they are one of the main noise sources in the reconstructed spectrum. In this work, we compare the signal-to-noise ratio performance of a prism-based S-FTS with that of a grating-based S-FTS. As a primary advantage, prisms give intrinsically lower diffused light than gratings. Furthermore, they do not have multiple diffracted orders, reducing thereafter the optical constraints on the instrumental baffling.

Super-Earths, M Dwarfs, and Photosynthetic Organisms: Habitability in the Lab.

In a few years, space telescopes will investigate our Galaxy to detect evidence of life, mainly by observing rocky planets. In the last decade, the observation of exoplanet atmospheres and the theoretical works on biosignature gasses have experienced a considerable acceleration. The most attractive feature of the realm of exoplanets is that 40% of M dwarfs host super-Earths with a minimum mass between 1 and 30 Earth masses, orbital periods shorter than 50 days, and radii between those of the Earth and Neptune (1-3.8 R⊕). Moreover, the recent finding of cyanobacteria able to use far-red (FR) light for oxygenic photosynthesis due to the synthesis of chlorophylls d and f, extending in vivo light absorption up to 750 nm, suggests the possibility of exotic photosynthesis in planets around M dwarfs. Using innovative laboratory instrumentation, we exposed different cyanobacteria to an M dwarf star simulated irradiation, comparing their responses to those under solar and FR simulated lights. As expected, in FR light, only the cyanobacteria able to synthesize chlorophyll d and f could grow. Surprisingly, all strains, both able or unable to use FR light, grew and photosynthesized under the M dwarf generated spectrum in a similar way to the solar light and much more efficiently than under the FR one. Our findings highlight the importance of simulating both the visible and FR light components of an M dwarf spectrum to correctly evaluate the photosynthetic performances of oxygenic organisms exposed under such an exotic light condition.

A New Remote Sensing-Based System for the Monitoring and Analysis of Growth and Gas Exchange Rates of Photosynthetic Microorganisms Under Simulated Non-Terrestrial Conditions.

Oxygenic photosynthetic microorganisms are a focal point of research in the context of human space exploration. As part of the bioregenerative life-support systems, they could have a key role in the production of breathable O2, edible biomasses and in the regeneration of CO2 rich-atmospheres and wastewaters produced by astronauts. The test of the organism's response to simulated physico-chemical parameters of planetary bodies could also provide important information about their habitability potential. It is believed that the success of future planetary and space missions will require innovative technologies, developed on the base of preliminary experiments in custom-made laboratory facilities. In this context, simulation chambers will play a pivotal role by allowing the growth of the microorganisms under controlled conditions and the evaluation in real-time of their biomass productivity and impact on atmosphere composition. We here present a system capable of addressing these requirements with high replicability and low costs. The setup is composed by three main parts: 1) a Star Light Simulator, able to generate different light intensities and spectra, including those of non-solar stars; 2) an Atmosphere Simulator Chamber where cultures of photosynthetic microorganisms can be exposed to different gas compositions; 3) a reflectivity detection system to measure from remote the Normalized Difference Vegetation Indexes (NDVI). Such a setup allows us to monitor photosynthetic microorganism's growth and gas exchange performances under selected conditions of light quality and intensity, temperature, pressure, and atmospheres simulating non-terrestrial environments. All parameters are detected by remote sensing techniques, thus without interfering with the experiments and altering the environmental conditions set. We validated the setup by growing cyanobacteria liquid cultures under different light intensities of solar illumination, collecting data on their growth rate, photosynthetic activity, and gas exchange capacity. We utilized the reflectivity detection system to measure the reflection spectra of the growing cultures, obtaining their relative NDVI that was shown to correlate with optical density, chlorophyll content, and dry weight, demonstrating the potential application of this index as a proxy of growth.

Comb-locked cavity ring-down spectrometer.

Extreme frequency accuracy and high sensitivity are obtained with a novel comb-locked cavity-ring-down spectrometer operating in the near-infrared from 1.5 to 1.63 µm. A key feature of our approach is the tight frequency locking of the probe laser to the comb, ensuring very high reproducibility and accuracy to the frequency axis upon scanning the comb repetition rate, as well as an efficient light injection into a length-swept high-finesse passive cavity containing the gas sample. Spectroscopic tests on the (30012) ← (00001) P14e line of CO2 at ∼1.57 µm demonstrate an accuracy of ∼17 kHz on the line center frequency in a Doppler broadening regime over the time scale of about 5 min, corresponding to four consecutive spectral scans of the absorption line. Over a single scan, which consists of 1500 spectral points over 75 s, the limit of detection is as low as 5.7 × 10(-11) cm(-1).

Noninvasive monitoring of gas in the lungs and intestines of newborn infants using diode lasers: feasibility study.

Preterm newborn infants have a high morbidity rate. The most frequently affected organs where free gas is involved are the lungs and intestines. In respiratory distress syndrome, both hyperexpanded and atelectatic (collapsed) areas occur, and in necrotizing enterocolitis, intramural gas may appear in the intestine. Today, these conditions are diagnosed with x-ray radiography. A bed-side, rapid, nonintrusive, and gas-specific technique for in vivo gas sensing would improve diagnosis. We report the use of noninvasive laser spectroscopy, for the first time, to assess gas content in the lungs and intestines of three full-term infants. Water vapor and oxygen were studied with two low-power diode lasers, illuminating the skin and detecting light a few centimeters away. Water vapor was easily detected in the intestines and was also observed in the lungs. The relatively thick chest walls of the infants prevented detection of the weaker oxygen signal in this study. However, results from a previous phantom study, together with scaling of the results presented here to the typical chest-wall thickness of preterm infants, suggest that oxygen also should be detectable in their lungs.