Combining Phi6 as a surrogate virus and computational large-eddy simulations to study airborne transmission of SARS-CoV-2 in a restaurant.

COVID-19 has highlighted the need for indoor risk-reduction strategies. Our aim is to provide information about the virus dispersion and attempts to reduce the infection risk. Indoor transmission was studied simulating a dining situation in a restaurant. Aerosolized Phi6 viruses were detected with several methods. The aerosol dispersion was modeled by using the Large-Eddy Simulation (LES) technique. Three risk-reduction strategies were studied: (1) augmenting ventilation with air purifiers, (2) spatial partitioning with dividers, and (3) combination of 1 and 2. In all simulations infectious viruses were detected throughout the space proving the existence long-distance aerosol transmission indoors. Experimental cumulative virus numbers and LES dispersion results were qualitatively similar. The LES results were further utilized to derive the evolution of infection probability. Air purifiers augmenting the effective ventilation rate by 65% reduced the spatially averaged infection probability by 30%-32%. This relative reduction manifests with approximately 15 min lag as aerosol dispersion only gradually reaches the purifier units. Both viral findings and LES results confirm that spatial partitioning has a negligible effect on the mean infection-probability indoors, but may affect the local levels adversely. Exploitation of high-resolution LES jointly with microbiological measurements enables an informative interpretation of the experimental results and facilitates a more complete risk assessment.

Soil microbial communities are shaped by vegetation type and park age in cities under cold climate.

Soil microbes play a key role in controlling ecosystem functions and providing ecosystem services. Yet, microbial communities in urban green space soils remain poorly characterized. Here we compared soil microbial communities in 41 urban parks of (i) divergent plant functional types (evergreen trees, deciduous trees and lawn) and (ii) different ages (constructed 10, ∼50 and >100 years ago). These microbial communities were also compared to those in 5 control forests in southern Finland. Our results indicate that, despite frequent disturbances in urban parks, urban soil microbes still followed the classic patterns typical of plant-microbe associations in natural environments: both bacterial and fungal communities in urban parks responded to plant functional groups, but fungi were under tighter control of plants than bacteria. We show that park age shaped the composition of microbial communities, possibly because vegetation in old parks have had a longer time to modify soil properties and microbial communities than in young parks. Furthermore, control forests harboured distinct but less diverse soil microbial communities than urban parks that are under continuous anthropogenic disturbance. Our results highlight the importance of maintaining a diverse portfolio of urban green spaces and plant communities therein to facilitate complex microbial communities and functions in urban systems.

Changes in TcpA gene frequency explain 2,4,6-trichlorophenol degradation in mesocosms.

Soils are often polluted by chlorophenols in timber production areas in the northern hemisphere. The tcpA gene encodes the first step of 2,4,6-trichlorophenol (246-TCP) degradation. We tested tcpA gene frequency in three natural pristine soils with different 246-TCP degradation capacity. Gene tcpA frequency increased more in spiked than non-spiked 10-L pails containing coniferous humus soil with high degradation capacity, in contrast to soils where degradation was slower. As the soil in each mesocosm originated from a spatially separate field plot, changes in tcpA gene frequency affected 246-TCP degradation over a range of soil origins. This indicates that the abundance of and changes in tcpA gene frequency could be utilized in estimating the efficacy of natural attenuation and biostimulation treatments in controlled conditions.

Hydrolase and plastic-degrading microbiota explain degradation of polyethylene terephthalate microplastics during high-temperature composting.

This research aims to explore the degradation properties of polyethylene terephthalate (PET) by PET hydrolase (WCCG) in high-temperature composting and its impact on microbial communities. PET degradation, composting parameters and microbial communities were assessed in 220 L sludge composters with PET and WCCG using high-throughput sequencing. Results showed that WCCG addition led to a deceleration of the humification process and a reduction in the relative abundance of thermophilic genera. Potential PET degrading microbiota, e.g. Acinetobacter, Bacillus, were enriched in the plastisphere in the composters where PET reduced by 26 % without WCCG addition. The external introduction of the WCCG enzyme to compost predominantly instigates a chemical reaction with PET, concurently curtailing the proliferation of plastic-degrading bacteria, leading to a 35 % degradation of PET. Both the WCCG enzyme and the microbiota associated with plastic-degradation showed the potential for reducing PET, offering a novel method for mitigating pollution caused by environmental microplastics.

Nutritional additives dominance in driving the bacterial communities succession and bioremediation of hydrocarbon and heavy metal contaminated soil microcosms.

Soil quality and microbial diversity are essential to the health of ecosystems. However, it is unclear how the use of eco-friendly natural additives can improve the quality and microbial diversity of contaminated soils. Herein, we used high-throughput 16 S rDNA amplicon Illumina sequencing to evaluate the stimulation and development of microbial diversity and concomitant bioremediation in hydrocarbon (HC) and heavy metal (HM)-rich waste disposal site soil when treated with meat and bone meal (MBM), cyclodextrin (Cdx), and MBM and cyclodextrin mixture (Cdx MBM) over a period of 3 months. Results showed that natural additive treatments significantly increased the soil bacterial diversity (higher Shannon index, Simpson index and evenness) in a time-dependent manner, with Cdx eliciting the greatest enhancement. The two additives influenced the bacterial community succession patterns differently. MBM, while it enhanced the enrichment of specific genera Chitinophaga and Terrimonas, did not significantly alter the total bacterial community. In contrast, Cdx or Cdx MBM promoted a profound change of the bacteria community over time, with the enrichment of the genera Parvibaculum, Arenimonas and unclassified Actinobacteria. These results provide evidence on the involvement of the two natural additives in coupling HC and HM bioremediation and bacterial community perturbations, and thus illustrates their potential application in ecologically sound bioremediation technologies for contaminated soils.

Bioremediation of contaminated soil and groundwater by in situ biostimulation.

Bioremediation by in situ biostimulation is an attractive alternative to excavation of contaminated soil. Many in situ remediation methods have been tested with some success; however, due to highly variable results in realistic field conditions, they have not been implemented as widely as they might deserve. To ensure success, methods should be validated under site-analogous conditions before full scale use, which requires expertise and local knowledge by the implementers. The focus here is on indigenous microbial degraders and evaluation of their performance. Identifying and removing biodegradation bottlenecks for degradation of organic pollutants is essential. Limiting factors commonly include: lack of oxygen or alternative electron acceptors, low temperature, and lack of essential nutrients. Additional factors: the bioavailability of the contaminating compound, pH, distribution of the contaminant, and soil structure and moisture, and in some cases, lack of degradation potential which may be amended with bioaugmentation. Methods to remove these bottlenecks are discussed. Implementers should also be prepared to combine methods or use them in sequence. Chemical/physical means may be used to enhance biostimulation. The review also suggests tools for assessing sustainability, life cycle assessment, and risk assessment. To help entrepreneurs, decision makers, and methods developers in the future, we suggest founding a database for otherwise seldom reported unsuccessful interventions, as well as the potential for artificial intelligence (AI) to assist in site evaluation and decision-making.

High-volume evacuation mitigates viral aerosol spread in dental procedures.

Dental healthcare personnel (DHCP) are subjected to microbe-containing aerosols and splatters in their everyday work. Safer work conditions must be developed to ensure the functioning of the healthcare system. By simulating dental procedures, we aimed to compare the virus-containing aerosol generation of four common dental instruments, and high-volume evacuation (HVE) in their mitigation. Moreover, we combined the detection of infectious viruses with RT-qPCR to form a fuller view of virus-containing aerosol spread in dental procedures. The air-water syringe produced the highest number of aerosols. HVE greatly reduced aerosol concentrations during procedures. The air-water syringe spread infectious virus-containing aerosols throughout the room, while other instruments only did so to close proximity. Additionally, infectious viruses were detected on the face shields of DHCP. Virus genomes were detected throughout the room with all instruments, indicating that more resilient viruses might remain infectious and pose a health hazard. HVE reduced the spread of both infectious viruses and viral genomes, however, it did not fully prevent them. We recommend meticulous use of HVE, a well-fitting mask and face shields in dental procedures. We advise particular caution when operating with the air-water syringe. Due to limited repetitions, this study should be considered a proof-of-concept report.

Molecular analysis of meso- and thermophilic microbiota associated with anaerobic biowaste degradation.

BACKGROUND: Microbial anaerobic digestion (AD) is used as a waste treatment process to degrade complex organic compounds into methane. The archaeal and bacterial taxa involved in AD are well known, whereas composition of the fungal community in the process has been less studied. The present study aimed to reveal the composition of archaeal, bacterial and fungal communities in response to increasing organic loading in mesophilic and thermophilic AD processes by applying 454 amplicon sequencing technology. Furthermore, a DNA microarray method was evaluated in order to develop a tool for monitoring the microbiological status of AD. RESULTS: The 454 sequencing showed that the diversity and number of bacterial taxa decreased with increasing organic load, while archaeal i.e. methanogenic taxa remained more constant. The number and diversity of fungal taxa increased during the process and varied less in composition with process temperature than bacterial and archaeal taxa, even though the fungal diversity increased with temperature as well. Evaluation of the microarray using AD sample DNA showed correlation of signal intensities with sequence read numbers of corresponding target groups. The sensitivity of the test was found to be about 1%. CONCLUSIONS: The fungal community survives in anoxic conditions and grows with increasing organic loading, suggesting that Fungi may contribute to the digestion by metabolising organic nutrients for bacterial and methanogenic groups. The microarray proof of principle tests suggest that the method has the potential for semiquantitative detection of target microbial groups given that comprehensive sequence data is available for probe design.

Natural additives contribute to hydrocarbon and heavy metal co-contaminated soil remediation.

A biological treatment method was tested in laboratory conditions for the removal of hydrocarbons contained in a waste disposal soil sample consisting of excavated sandy soil from a former fueling station. Two fractions of hydrocarbons were quantified by GC-FID: diesel (C10-C21) and lubricant oil (C22-C40). Meat and bone meal (MBM, 1% w/w) was used as a bio-stimulant agent for soil organisms. Cyclodextrin, an oligosaccharide produced from starch by enzymatic conversion, was also used to assess its ability to improve the bioavailability/biodegradability of hydrocarbons in the soil. Parameters such as temperature, pH, water content and aeration (O2 availability) were monitored and optimized to favor degradation processes. Two different experimental tests were prepared: one to measure the degradation of hydrocarbons; the other to monitor the mobility of some elements in the soil and in the leachate produced by watering with tap water. Soil samples treated with MBM and cyclodextrin showed, over time, a greater removal of the more persistent hydrocarbon fraction (lubricant oil). MBM-treated soils underwent a faster hydrocarbon removal kinetic, especially in the first treatment period. However, the final hydrocarbon concentrations are comparable in all treatments, including control. Over time, the effect of cyclodextrin on hydrocarbon degradation seemed to be relevant. MBM-treated soils sequestered lead in the very first weeks. These results highlight the intrinsic capacity of soil, and its indigenous microbial communities, to degrade petroleum hydrocarbons and suggest that MBM-induced bioremediation is a promising, environmentally friendly technology which should be considered when dealing with hydrocarbon/heavy metal co-contaminated soils.

Meat and bone meal stimulates microbial diversity and suppresses plant pathogens in asparagus straw composting.

Meat and bone meal (MBM), as slaughterhouse waste, is a potential biostimulating agent, but its efficiency and reliability in composting are largely unknown. To access the MBM application to the composting process of asparagus straw rice, we followed the composting process for 60 days in 220-L composters and another 180 days in 20-L buckets in treatments applied with MBM or urea. The microbial succession was investigated by high-throughput sequencing. Compared with urea treatments, MBM addition stabilized pH and extended the thermophilic phase for 7 days. The germination index of MBM treatments was 24.76% higher than that of urea treatments. MBM also promoted higher microbial diversity and shifted community compositions. Organic matter and pH were the most significant factors that influence the bacterial and fungal community structure. At the genus level, MBM enriched relative abundances of organic matter-degrading bacteria (Alterococcus) and lignocellulose-degrading fungi (Trichoderma), as well as lignocellulolytic enzyme activities. Notably, MBM addition decreased sum abundances of plant pathogenic fungi of Phaeoacremonium, Acremonium, and Geosmithia from 17.27 to 0.11%. This study demonstrated the potential of MBM as an effective additive in asparagus straw composting, thus providing insights into the development of new industrial aerobic fermentation.

Dataset on bio-stimulation experiments for the removal of hydrocarbons and the monitoring of certain elements in a contaminated soil.

Meat and Bone Meal (MBM) and ß-cyclodextrin were added to a soil sample co-contaminated by hydrocarbons (diesel fraction C10-C21 and lubricant oil fraction C22-C40) and heavy metals to promote soil remediation. The pilot study was conducted in the laboratory, maintaining optimal conditions (i.e., temperature, pH, water content, soil aeration) to facilitate hydrocarbon biodegradation. Two different experimental tests were prepared: one for the analysis of hydrocarbons in soil, the other to monitor the dynamics of some elements of interest. For the first test, the two hydrocarbon fractions in the soil were quantified separately by GC-FID, following the ISO 16703:2004(E) standard protocol. Sampling and analysis were done every two weeks, for three consecutive months. For the second test (dynamics of certain elements in the soil), soil and leachate samples were analyzed by ICP-MS after appropriate pretreatment steps.

In situ electrokinetic (EK) remediation of the total and plant available cadmium (Cd) in paddy agricultural soil using low voltage gradients at pilot and full scales.

Electrokinetic (EK) remediation has been widely studied at laboratory scales. However, field-scale research is far less. In this study, a 14-day EK remediation was carried out, in a field pilot (4 m2) test and a full-scale (200 m2) application for the first time, in a cadmium (Cd) contaminated paddy agricultural field near a mining area. A low voltage of 20 V was applied at both scales; voltage gradient was 20 V m-1 and 4 V m-1 at the pilot and full scales, respectively. Samples were taken from near the anode and cathode, and in the middle of the electric field, in the soil layers 0-10 cm, 10-20 cm, and 40-50 cm. After the EK remediation, a significant portion of the total Cd was removed in all the layers at the pilot scale, by 87%, 72%, and 54% from the top down, but only in the 0-10 cm layer at the full scale by 74%. As for the plant available (exchangeable and soluble) Cd, significant removal (64%) was only observed in the 0-10 cm layer at the pilot scale. The percentage reduction of the electrical conductivity and removal efficiency of the total Cd was higher near the anode than the cathode. The soil pH was elevated near the cathode but stayed below pH 6 due to the sufficient supply of lactic acid. After the EK remediation, the concentration of the total Cd dropped below the hazard threshold, i.e. 0.4 mg (kg dry wt soil)-1 for agricultural paddy fields in China. A total energy of 2 kW·h and 0.6 kW·h was consumed at the pilot and full scales, respectively. This study showed a successful in situ EK remediation of Cd contaminated paddy agricultural soil, especially in the surface layer, with low voltage and energy demand.

In situ bioremediation of Fenton's reaction-treated oil spill site, with a soil inoculum, slow release additives, and methyl-ß-cyclodextrin.

A residential lot impacted by spills from a leaking light heating oil tank was treated with a combination of chemical oxidation and bioremediation to avoid technically challenging excavation. The tank left emptied in the ground was used for slow infiltration of the remediation additives to the low permeability, clayey soil. First, hydrogen peroxide and citrate chelate was added for Fenton's reaction-based chemical oxidation, resulting in a ca. 50% reduction from the initial 25,000 mg/kg average oil concentration in the soil below the tank. Part of this was likely achieved through mobilization of oily soil into the tank, which was beneficial in regards to the following biological treatment. By first adding live bacteria in a soil inoculum, and then oxygen and nutrients in different forms, an approximately 90% average reduction was achieved. To further enhance the effect, methyl-ß-cyclodextrin surfactant (CD) was added, resulting finally in a 98% reduction from the initial average level. The applicability of the surfactant was based on laboratory-scale tests demonstrating that CD promoted oil degradation and, unlike pine soap, was not utilized by the bacteria as a carbon source, and thus inhibiting degradation of oils regardless of the positive effect on biological activity. The effect of CD on water solubility for different hydrocarbon fractions was tested to serve as the basis for risk assessment requirements for authorizing the use of the surfactant at the site.

Habitat Elevation Shapes Microbial Community Composition and Alter the Metabolic Functions in Wild Sable (Martes zibellina) Guts.

In recent decades, wild sable (Carnivora Mustelidae Martes zibellina) habitats, which are often natural forests, have been squeezed by anthropogenic disturbances such as clear-cutting, tilling and grazing. Sables tend to live in sloped areas with relatively harsh conditions. Here, we determine effects of environmental factors on wild sable gut microbial communities between high and low altitude habitats using Illumina Miseq sequencing of bacterial 16S rRNA genes. Our results showed that despite wild sable gut microbial community diversity being resilient to many environmental factors, community composition was sensitive to altitude. Wild sable gut microbial communities were dominated by Firmicutes (relative abundance 38.23%), followed by Actinobacteria (30.29%), and Proteobacteria (28.15%). Altitude was negatively correlated with the abundance of Firmicutes, suggesting sable likely consume more vegetarian food in lower habitats where plant diversity, temperature and vegetation coverage were greater. In addition, our functional genes prediction and qPCR results demonstrated that energy/fat processing microorganisms and functional genes are enriched with increasing altitude, which likely enhanced metabolic functions and supported wild sables to survive in elevated habitats. Overall, our results improve the knowledge of the ecological impact of habitat change, providing insights into wild animal protection at the mountain area with hash climate conditions.

Bacterial diversity at different stages of the composting process.

BACKGROUND: Composting is an aerobic microbiological process that is facilitated by bacteria and fungi. Composting is also a method to produce fertilizer or soil conditioner. Tightened EU legislation now requires treatment of the continuously growing quantities of organic municipal waste before final disposal. However, some full-scale composting plants experience difficulties with the efficiency of biowaste degradation and with the emission of noxious odours. In this study we examine the bacterial species richness and community structure of an optimally working pilot-scale compost plant, as well as a full-scale composting plant experiencing typical problems. Bacterial species composition was determined by isolating total DNA followed by amplifying and sequencing the gene encoding the 16S ribosomal RNA. RESULTS: Over 1500 almost full-length 16S rRNA gene sequences were analysed and of these, over 500 were present only as singletons. Most of the sequences observed in either one or both of the composting processes studied here were similar to the bacterial species reported earlier in composts, including bacteria from the phyla Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria and Deinococcus-Thermus. In addition, a number of previously undetected bacterial phylotypes were observed. Statistical calculations estimated a total bacterial diversity of over 2000 different phylotypes in the studied composts. CONCLUSIONS: Interestingly, locally enriched or evolved bacterial variants of familiar compost species were observed in both composts. A detailed comparison of the bacterial diversity revealed a large difference in composts at the species and strain level from the different composting plants. However, at the genus level, the difference was much smaller and illustrated a delay of the composting process in the full-scale, sub-optimally performing plants.

Reduction of odorants in swine manure by carbohydrate and bacterial amendments.

Malodors from pig manure storage pits are a problem lacking a cost-efficient solution particularly for small pig (Sus domestica) farms. The objective of this study was to reduce emissions of noxious odorants by changing the conditions in the pig manure to favor an altered microbial community. Sugars (maltose, lactose, and saccharose), carbohydrate-rich waste (maltose syrup and wheat flour), and bacterial amendments (Lactobacillus plantarum and L. amylophilus) were tested for their effect on manure pH, bacterial community, and gaseous emissions. In laboratory experiments, a permanent pH reduction was achieved with all carbohydrates when added to a 5% final concentration. Maltose significantly decreased emissions of sulfur-containing compounds, whereas bacterial amendments had little effect on odorants. Lactobacilli were found in manure receiving carbohydrate amendments alone, but Clostridiales, Bacteroidetes, and Enterobacteriaceae were observed in all treatments (including the control). At the mesocosm (60 L) level, maltose syrup and wheat flour amendments caused clear pH reductions in manure, whereas L. plantarum and L. amylophilus had no additional effect. The addition of maltose syrup and wheat flour to a manure storage pit (600 m(3)) lowered the pH slightly. When the manure was spread onto the fields, the observed reduction in odor was found to be reversible, and the release of malodors was delayed rather than eliminated. We conclude that these methods require further development to produce a reliable technical application.

Simulation of Microbial Response to Accidental Diesel Spills in Basins Containing Brackish Sea Water and Sediment.

The brackish Baltic Sea is under diesel oil pollution risk due to heavy ship traffic. The situation is exasperated by densely distributed marinas and a vigorous although seasonal recreational boating. The seasonality and physical environmental variations hamper the monitoring of microbial communities in response to diesel oil spills. Hence, an 8-week simulation experiment was established in metal basins (containing 265 L sea water and 18 kg quartz sand or natural shore sand as the littoral sediment) to study the effect of accidental diesel oil spills on microbial communities. Our results demonstrated that microbial communities in the surface water responded to diesel oil contamination, whereas those in the littoral sediment did not, indicating that diesel oil degradation mainly happened in the water. Diesel oil decreased the abundance of bacteria and fungi, but increased bacterial diversity in the water. Time was the predominant driver of microbial succession, attributable to the adaption strategies of microbes. Bacteria were more sensitive to diesel oil contamination than fungi and archaea. Diesel oil increased relative abundances of bacterial phyla, Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Flavobacteriia and Cytophagia, and fungal phylum Ascomycota in the surface water. Overall, this study improves the understanding of the immediate ecological impact of accidental diesel oil contamination, providing insights into risk management at the coastal area.

Yard vegetation is associated with gut microbiota composition.

Gut microbes play an essential role in the development and functioning of the human immune system. A disturbed gut microbiota composition is often associated with a number of health disorders including immune-mediated diseases. Differences in host characteristics such as ethnicity, living habit and diet have been used to explain differences in the gut microbiota composition in inter-continental comparison studies. As our previous studies imply that daily skin contact with organic gardening materials modify gut microflora, here we investigated the association between living environment and gut microbiota in a homogenous western population along an urban-rural gradient. We obtained stool samples from 48 native elderly Finns in province Häme in August and November 2015 and identified the bacterial phylotypes using 16S rRNA Illumina MiSeq sequencing. We assumed that yard vegetation and land cover classes surrounding homes explain the stool bacterial community in generalized linear mixed models. Diverse yard vegetation was associated with a reduced abundance of Clostridium sensu stricto and an increased abundance of Faecalibacterium and Prevotellaceae. The abundance of Bacteroides was positively and strongly associated with the built environment. Exclusion of animal owners did not alter the main associations. These results suggest that diverse vegetation around homes is associated with health-related changes in gut microbiota composition. Manipulation of the garden diversity, possibly jointly with urban planning, is a promising candidate for future intervention studies that aim to maintain gut homeostasis.

Fenton's reaction-based chemical oxidation in suboptimal conditions can lead to mobilization of oil hydrocarbons but also contribute to the total removal of volatile compounds.

Fenton's reaction-based chemical oxidation is in principle a method that can be utilized for all organic fuel residues thus making it a potential all-purpose, multi-contaminant, in situ application for cases in which storage and distribution of different types of fuels have resulted in contamination of soil or groundwater. Since peroxide breakdown reactions are also expected to lead to a physical transport of the target compound, this secondary physical removal, or rebound concentrations related to it, is prone to be affected by the chemical properties of the target compound. Also, since soil conditions are seldom optimal for Fenton's reaction, the balance between chemical oxidation and transport may vary. In this study, it was found that, with a high enough hydrogen peroxide concentration (5 M), methyl tert-butyl ether-spiked groundwater could be treated even under suboptimal conditions for chemical mineralization. In these cases, volatilization was not only contributing to the total removal but also leading to rebound effects similar to those associated with air sparging techniques. Likewise for diesel, temporal transport from soil to the aqueous phase was found to lead to false positives that outweighed the actual remediation effect through chemical mineralization.

Influence of long term nitrogen limitation on lipid, protein and pigment production of Euglena gracilis in photoheterotrophic cultures.

Nitrogen limitation is considered a good strategy for enhancement of algal lipid production while conversely N repletion has been shown to result in biomass rich in proteins. In this study, the influence of long-term N limitation on Euglena gracilis fatty acid (FA), protein, chlorophyll a, and carotenoid concentrations was studied in N limited cultures. Biomass composition was analyzed from three-time points from N starved late stationary phase cultures, exposed to three different initial N concentrations in the growth medium. Total lipid content increased under N limitation in ageing cultures, but the low N content and prolonged cultivation time resulted in the formation of a high proportion of saturated FAs. Furthermore, growth as well as the production of proteins, chlorophyll a and carotenoids were enhanced in higher N concentrations and metabolism of these cellular components stayed stable during the stationary growth phase. Our findings showed that a higher N availability and a shorter cultivation time is a good strategy for efficient E. gracilis biomass production, regardless of whether the produced biomass is intended for maximal recovery of polyunsaturated FAs, proteins, or photosynthetic pigments. Additionally, we showed an increase of neoxanthin, ß-carotene, and diadinoxanthin as a response to higher N availability.