Cardiac fat adipocytes: An optimized protocol for isolation of ready-to-use mature adipocytes from human pericardial adipose tissue.

A better understanding of the pathophysiology of cardiac fat depots is crucial to describe their role in the development of cardiovascular diseases. To this end, we have developed a method to isolate mature fat cells from the pericardial adipose tissue (PAT), the most accessible cardiac fat depot during cardiac surgery. Using enzymatic isolation, we were able to successfully obtain mature fat cells together with the corresponding cells of the stromal vascular fraction (SVF). We subjected the PAT adipocytes to thorough morphological and molecular characterization, including detailed fatty acid profiling, and simultaneously investigated their reactivity to external stimuli. Our approach resulted in highly purified fat cells with sustained viability for up to 72 h after explantation. Remarkably, these adipocytes responded to multiple challenges, including pro-inflammatory and metabolic stimuli, indicating their potential to trigger a pro-inflammatory response and modulate endothelial cell behavior. Furthermore, we have created conditions to maintain whole PAT in culture and preserve their viability and reactivity to external stimuli. The efficiency of cell recovery combined with minimal dedifferentiation underscores the promise for future applications as a personalized tool for screening and assessing individual patient responses to drugs and supplements or nutraceuticals.

Investigating the impact of wet rendering (solventless method) on PUFA-rich oil from catfish (Clarias magur) viscera.

Catfish (Clarias magur) is a popular freshwater fish food worldwide. The processing of this fish generates a significant amount of waste, mainly in the form of viscera, which constitutes around 10-12% of the fish's total weight. This study was focused on extracting polyunsaturated fatty acid (PUFA)-rich oil from catfish viscera, aiming to enhance the extraction process and make the production of oil and handling of fish byproducts more cost-effective. The wet reduction method, a solvent-free approach, was used for extraction, with yield optimization done via the Box-Behnken design. The resulting oil was evaluated for its oxidative quality and chemical characteristics. The optimal conditions for the wet rendering process were as follows: viscera to water ratio, 1:0.5 (w/v); temperature, 90℃; and time, 20 min, yielding 12.40 g/100 g of oil. The oil extracted under optimal wet rendering conditions had quality and oxidative stability comparable to solvent extraction and fewer secondary oxidation compounds. This oil had a higher PUFA content, specifically a 4:1 ratio of omega 6 to omega 3. Such oil, derived from catfish viscera, is suitable for the food industry due to its solvent-free extraction method.

Distribution and Level of Bioactive Monoacylglycerols in 12 Marine Microalgal Species.

Microalgae are currently considered an attractive source of highly valuable metabolites potentially exploitable as anticancer agents, nutraceuticals and cosmeceuticals and for bioenergy purposes. Their ease of culturing and their high growth rates further promote their use as raw material for the production of specialty products. In the present paper, we focused our attention on specific glycerol-based lipid compounds, monoacylglycerols (MAGs), which displayed in our previous studies a selective cytotoxic activity against the haematological U-937 and the colon HCT-116 cancer cell lines. Here, we performed a quali/quantitative analysis of MAGs and total fatty acids (FAs) along with a profiling of the main lipid classes in a panel of 12 microalgal species, including diatoms and dinoflagellates. Our results highlight an inter- and intraspecific variability of MAG profile in the selected strains. Among them, Skeletonema marinoi (strain FE7) has emerged as the most promising source for possible biotechnological production of MAGs.

Soil nitrogen availability and microbial carbon use efficiency are dependent more on chemical fertilization than winter drought in a maize-soybean rotation system.

Soil nitrogen (N) availability is one of the limiting factors of crop productivity, and it is strongly influenced by global change and agricultural management practices. However, very few studies have assessed how the winter drought affected soil N availability during the subsequent growing season under chemical fertilization. We conducted a field investigation involving snow removal to simulate winter drought conditions in a Mollisol cropland in Northeast China as part of a 6-year fertilization experiment, and we examined soil physicochemical properties, microbial characteristics, and N availability. Our results demonstrated that chemical fertilization significantly increased soil ammonium and total N availability by 42.9 and 90.3%, respectively; a combined winter drought and fertilization treatment exhibited the highest soil N availability at the end of the growing season. As the growing season continued, the variation in soil N availability was explained more by fertilization than by winter drought. The Mantel test further indicated that soil Olsen-P content and microbial carbon use efficiency (CUE) were significantly related to soil ammonium availability. A microbial community structure explained the largest fraction of the variation in soil nitrate availability. Microbial CUE showed the strongest correlation with soil N availability, followed by soil available C:P and bacteria:fungi ratios under winter drought and chemical fertilization conditions. Overall, we clarified that, despite the weak effect of the winter drought on soil N availability, it cannot be ignored. Our study also identified the important role of soil microorganisms in soil N transformations, even in seasonally snow-covered northern croplands.

Analysis of commercial fetal bovine serum (FBS) and its substitutes in the development of cultured meat.

Fetal bovine serum (FBS) is an extremely important culture growth supplement, accounting for approximately 60 % of cell-culture-media costs; therefore, lowering FBS-acquisition costs for the industrialization of cultured meat is imperative. This study attempted to produce an FBS substitute using discarded livestock by-products, with particular focus on formulating a product with a composition similar to that of FBS to improve effectiveness. However, to date, no study has precisely analyzed the commercial components of FBS, and this study is the first to compare the chemical composition of FBS and commercially available horse serum purchased from the United States or Europe with that of FBS substitutes developed by our team. This study analyzed the chemical composition of the FBS products purchased by our team over the past 3 years via blood, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and independent composition analyses. While the composition and quality of commercial FBS products are known to vary, the FBS composition of our purchased products was relatively uniform regardless of company, brand, or country of origin. In contrast, FBS substitutes obtained from three major livestock species (cattle, pig, and chicken) clearly exhibited differences in composition, a phenomenon that was also observed upon comparing with FBS as well as among different species. Therefore, to replace commercial FBS entirely, the production of a proportionately effective substitute product comprising an equal or similar composition is required, and the results of this study can be a steppingstone to achieving this. In addition, FBS substitutes manufactured using inexpensive slaughter by-products as raw materials are expected to ultimately reduce the unit cost of cultured meat production.

Toward Renewable-Based Prebiotics from Woody Biomass: Potential of Tailored Xylo-Oligosaccharides Obtained by Enzymatic Hydrolysis of Beechwood Xylan as a Prebiotic Feed Supplement for Young Broilers.

Although antibiotic resistance emerges naturally, this process has been accelerated by the worldwide overuse and misuse of antibiotics. It is essential to find effective alternatives in the broiler industry to improve poultry health while maintaining production efficiency and product safety. In this study, we aimed to evaluate a potential alternative: wood-derived xylo-oligosaccharides (XOS). The objective of this research was to investigate the potential of XOS prepared using enzymatic hydrolysis of beechwood xylan as a prebiotic feed supplement for broilers. A pilot study was conducted to explore the optimal XOS fraction profile by in vitro fermentation. Subsequently, a semi-continuous enzyme membrane reactor was used, allowing for the production of tailored XOS in large quantities. Given the strong bidirectional relationship between intestinal health, nutrition, and intestinal microbiota composition in broilers, an in vivo experiment was performed to explore the potential of XOS as a prebiotic feed supplement by investigating growth performance, feed conversion ratio, caecal short and medium chain fatty acid (SCFA and MCFA) concentration, and microbiological composition of the caecal content. Results from the pilot study indicated that higher enzyme concentrations in the hydrolysis process yield a product that leads to a higher total SCFA and MCFA- and butyric acid production during in vitro fermentation by caecal bacteria. Supplementation of the tailored XOS to the broiler diet (day 1 (d1)-d8 0.13% wt/wt XOS, d9-d15 0.32% XOS) resulted in higher Bifidobacterium counts, beneficial to the health of birds, on d11 and d15.

Influence of solvent-free extraction of fish oil from catfish (Clarias magur) heads using a Taguchi orthogonal array design: A qualitative and quantitative approach.

This study aimed to efficiently utilize catfish heads, enhancing the oil extraction process while improving the cost-effectiveness of fish byproduct management. The study employed the wet rendering method, a solvent-free approach, utilizing a two-factor Taguchi orthogonal array design to identify critical parameters for optimizing oil yield and ensuring high-quality oil attributes. The extraction temperature (80-120°C) and time (5-25 min) were chosen as variables in the wet rendering process. Range analysis identified the extraction time as a more significant (p < 0.05) factor for most parameters, including oil yield, oil recovery, acid value, free fatty acids, peroxide value, and thiobarbituric acid reactive substances. The extraction temperature was more significant (p < 0.05) for oil color. Consequently, the wet rendering method was optimized, resulting in an extraction temperature of 80°C and an extraction time of 25 min, yielding the highest oil yield. This optimized wet rendering process recovered 6.37 g/100 g of oil with an impressive 54.16% oil recovery rate, demonstrating comparable performance to traditional solvent extraction methods. Moreover, Fourier transfer infrared spectra analysis revealed distinct peaks associated with triacylglycerols and polyunsaturated fatty acids (PUFA). The oil recovered under optimized conditions contained higher levels of PUFA, including oleic acid (189.92 µg/g of oil), linoleic acid (169.92 µg/g of oil), eicosapentaenoic acid (17.41 µg/g of oil), and docosahexaenoic acid (20.82 µg/g of oil). Volatile compound analysis revealed lower levels of secondary oxidation compounds under optimized conditions. This optimized wet rendering method offers practical advantages in terms of cost-efficiency, sustainability, reduced environmental impact, and enhanced oil quality, making it an attractive option for the fish processing industries. Future research possibilities may include the purification of the catfish head oil and its application in the food and pharmaceutical industries.

Equal bioavailability of omega-3 PUFA from Calanus oil, fish oil and krill oil: A 12-week randomized parallel study.

The bioavailability of long-chain omega-3 polyunsaturated fatty acids (n3 PUFA) can be affected by the form in which they are bound. An alternative source of n3 PUFA is Calanus finmarchicus oil (CO), which, unlike fish oil (FO) and krill oil (KO), contains fatty acids primarily bound as wax esters. Recent studies have shown that n3 PUFA from CO are bioavailable to humans, but CO has not been compared to other marine oils such as FO or KO. Therefore, the aim of this study was to investigate the influence of 12 weeks supplementation with CO, FO and KO on the long-term n3 PUFA status in healthy volunteers. The Omega-3 Index (O3I), defined as red blood cell EPA + DHA content as a percentage of total identified fatty acids, was used as a measure to assess n3 PUFA status. Sixty-two participants (mean ± standard deviation [SD] age: 29.7 ± 8.43 years) completed the randomized parallel group study (CO group: n = 21, 4 capsules/day, EPA + DHA dose: 242 mg/day; FO group: n = 22, 1 capsule/day, EPA + DHA dose: 248 mg/day; KO group: n = 19, 2 capsules/day, EPA + DHA dose: 286 mg/day). At baseline, the three groups showed comparable (mean ± SD) O3I values (CO: 5.13 ± 1.12%, FO: 4.90 ± 0.57%, KO: 4.87 ± 0.77%). The post-interventional (mean ± SD) O3I increase was comparable between the three groups (CO: 1.09 ± 0.55%; FO: 1.0 ± 0.53%; KO: 1.15 ± 0.65%, all p < 0.001). The study confirms that CO can increase the n3 PUFA status comparable to FO and KO and is therefore an alternative marine source of bioavailable n3 PUFA, especially with regard to sustainability.

Plant-soil interactions in the native range of two congeneric species with contrasting invasive success.

The aim of this study was to compare plant-soil interactions in the native range of two congeneric European species differing in their invasive success in the world: a globally invasive Cirsium vulgare and non-invasive C. oleraceum. We assessed changes in soil nutrients and soil biota following soil conditioning by each species and compared performance of plants grown in self-conditioned and unconditioned soil, from which all, some or no biota was excluded. The invasive species depleted more nutrients than the non-invasive species and coped better with altered nutrient levels. The invasive species had higher seedling establishment which benefited from the presence of unconditioned biota transferred by soil filtrate. Biomass of both species increased in soil with self-conditioned soil filtrate and decreased in soil with self-conditioned whole-soil inoculum compared to unconditioned filtrate and inoculum. However, the increase was smaller and the decrease greater for the invasive species. The invasive species allocated less biomass to roots when associated with harmful biota, reducing negative effects of the biota on its performance. The results show that in the native range the invasive species is more limited by self-conditioned pathogens and benefits more from unconditioned mutualists and thus may benefit more from loss of effectively specialized soil biota in a secondary range. Our study highlights the utility of detailed plant-soil feedback research in species native range for understanding factors regulating species performance in their native range and pinpointing the types of biota involved in their regulation.

Vascular plant and cryptogam abundance as well as soil chemical properties shape microbial communities in the successional gradient of glacier foreland soils.

In the glacier forelands, microbes play a fundamental role in soil development and shaping the vegetation structure. Such ecosystems represent various stages of soil development and are, therefore, an excellent place to study the interrelationship between soil, plants, and microorganisms. The aim of the study was to assess the effects of vegetation and soil physicochemical properties developing after glacier retreat on soil microbial communities. Specifically, abundance, species richness and the composition of arbuscular mycorrhizal fungi (AMF), as well as microbial biomass and community structure in soils were compared between plots established in 800-meter transects of three glacier forelands in northern Sweden. The cover of vascular plants and cryptogams, soil C content, AMF spore density and species richness, AMF biomass indicators, total microbial biomass, and bacterial phospholipid fatty acids (PLFA) were significantly and positively related to the distance from the glacier terminus. On the other hand, macronutrient concentrations and pH decreased along with increasing distance. No significant impact of the distance from the glacier terminus on the ratio fungal/bacterial PLFA was observed. Moreover, we found a significant effect of both glacier and the distance from the glacier terminus on the microbial community structure. AMF species richness and spore density in the glacier forelands were generally low, which is probably due to a limited supply of inoculum in primary successional ecosystems. Most microbial biochemical markers and AMF parameters were positively associated with the number of arbuscular mycorrhizal plant species and vascular plant and lichen cover as well as C content in soil, whereas negatively with soil macronutrients and pH. This could be related to an increase in plant cover and a decrease in soil nutrient levels as plant succession progresses. Our results showed that vegetation, soil C content, and microbial communities are interlinked and exhibit concordant patterns along successional gradients.

Microscale supercritical fluid extraction combined with supercritical fluid chromatography and proton-transfer-reaction ionization time-of-flight mass spectrometry for a magnitude lower limit of quantitation of lipophilic compounds.

The application of proton transfer ionization reaction mass spectrometry (PTR MS) combined with microscale supercritical fluid extraction (SFE) and supercritical fluid chromatography (SFC) aiming to quantitate single-cell fatty acid analysis levels was investigated. Using a microscale extraction vessel, the obtained low limits of quantitation (LLOQs) of arachidonic acid and arachidic acid were 1.2 and 2.7 fmol, respectively, by using less than 1 µL of sample on stainless steel frit. A series of phthalate, vitamin K1, and α-tocopherol were also tested, and the LLOQ was less than one femtomole for phthalate and 35 and 13 fmol for vitamin K1 and α-tocopherol, respectively. A microliter portion of SFE extracts was introduced into the SFC column by split injection, improving the reproducibility of the chromatography and separation efficiency. The method in the present study has great potential to quantitate lipophilic molecules on the nanogram scale of a sample without complex preparation procedures.

Indictors of wetland health improve following small-scale ecological restoration on private land.

Globally wetlands are imperilled and restoring these highly productive and biodiverse ecosystems is key to regaining their lost function and health. Much of the fertile, low-lying land that was historically wetland is now farmed, so privately-owned locations play critical roles in regaining space for wetlands. However, wetland restoration on private property is often small-scale and supported by minimal funding and expertise. Little is known about what these efforts achieve, and what contexts facilitate the greatest gains in wetland health. Using a paired plot design for 18 restored and 18 unrestored wetlands, we aimed to understand changes in wetland health following restoration on private property. We characterised plant and microbial communities and soil characteristics following wetland restoration and explored how environmental settings of restored wetlands related to the clustering of wetland health indicators. We found that all indicators of wetland health significantly increased following restoration except for the ratio of Gram negative to Gram positive bacteria. Restoration enhanced plant alpha and beta diversity, adding ~13 native plant species per plot. Soils in restored wetlands contained 20% more organic matter, and 25% more microbial biomass, which was driven by an increased abundance of fungi. Restoration reduced soil bulk density by 0.19 g-1 cm3 and Olsen Phosphorus by 23%. These effects on soil physical characteristics and microbial communities were strongest in the wettest locations. Restored wetlands clustered into three main groups based on indicators of wetland health. Hydrological flow explained the clustering of wetlands, with riverine wetlands exhibiting greater indicators of recovery than depressional wetlands, suggesting that hydrological flow may influence post-restoration recovery. Overall, this study shows that small-scale wetland restoration on private land improved wetland health, providing evidence that it can be an effective use of marginal agricultural land.

Spatial and temporal variations in salt marsh microorganisms of the Wadden Sea.

Salt marshes exist at the interface of the marine and the terrestrial system. Shore height differences and associated variations in inundation frequency result in altered abiotic conditions, plant communities, and resource input into the belowground system. These factors result in three unique zones, the upper salt marsh (USM), the lower salt marsh (LSM), and the pioneer zone (PZ). Marine detritus, such as micro- and macroalgae, is typically flushed into the PZ daily, with storm surges moving both salt marsh detritus and marine detritus into higher salt marsh zones. Microbial assemblages are essential for the decomposition of organic matter and have been shown to sensitively respond to changes in abiotic conditions such as oxygen supply and salinity. However, temporal and spatial dynamics of microbial communities of Wadden Sea salt marshes received little attention. We investigated the dynamics of soil microbial communities across horizontal (USM, LSM, and PZ), vertical (0-5 and 5-10-cm sediment depth), and temporal (spring, summer, and autumn) scales in the Wadden Sea salt marsh of the European North Atlantic coast using phospholipid fatty acid (PLFA) analysis. Our results show strong spatial dynamics both among salt marsh zones and between sediment depths, but temporal dynamics to be only minor. Despite varying in space and time, PLFA markers indicated that bacteria generally were the dominant microbial group across salt marsh zones and seasons, however, their dominance was most pronounced in the USM, whereas fungal biomass peaked in the LSM and algal biomass in the PZ. Only algal markers and the stress marker monounsaturated to saturated fatty acid ratio responded to seasonality. Overall, therefore, the results indicate remarkable temporal stability of salt marsh microbial communities despite strong variability in abiotic factors.

[Characteristics of Microbial Utilization for Crop Residue-Derived C in Paddy and Upland Soils].

Two typical subtropical agricultural soils, a flooded paddy soil and its adjacent upland, were collected and then incubated with or without 13C-labeled crop residue (maize straw) for 40 days. During the incubation, the mineralization rate of the crop residue was monitored, and the 13C incorporated into fungal and bacterial phospholipid fatty acid (PLFA) was quantified. At the early stage (0.25-1 days), the mineralization rate of crop residue was faster in paddy soil than that in upland soil, whereas the opposite trend was observed from 2 to 20 days. At the late stage (21-40 days), the mineralization rate was similar in both soils. At the end of incubation, 11% of the total crop residue was mineralized in paddy soil, which was about half of that in upland soil (20%). Although paddy soil had a higher amount of microbial biomass (indicated by total PLFA), the total amounts of 13C-PLFA were comparable in both soils, and the enrichment ratio (proportion of 13C to total C in PLFA) was lower in paddy soil than that in upland soil. This indicated that the microbial community in paddy soil was less active in the uptake of crop residue C than that in upland soil. During the incubation, the residue-derived 13C was mainly distributed in bacterial PLFA (up to 86% of total 13C-PLFA, including 59% in gram-positive and 27% in gram-negative bacteria) in paddy soil, and up to 75% of total 13C-PLFA distributed in fungal PLFAs was in upland soil. Thus, bacteria dominated the utilization of crop residue in paddy soil versus fungi in upland soil. Compared with that in upland soil, the microbial activity was suppressed in the anaerobic condition caused by flooding in paddy soil, with a stronger inhibition of fungi than bacteria. Considering the discrepancies of life strategies and necromass turnover between bacteria and fungi, the different dominant microbial groups in the utilization of crop residue in water-logged and well-drained conditions could lead to the distinct accumulation and stabilization of microbial-derived organic matter in paddy and upland soils.

Herbaceous plant species support soil microbial performance in deciduous temperate forests.

Although herbaceous plant layer may contribute significantly to plant diversity and nutrient turnover, its effects on the soil environment in forest ecosystems remain largely unexplored. In this study, we compared the effects of mono-dominant and multi-species assemblages of herb plants on soil physicochemical and microbial properties in two temperate deciduous (beech and riparian) forests. We hypothesized that the presence of herbaceous plants would increase microbial activity and biomass, and nutrient availability in soil when compared to bare soil. This increase would be the highest in multi-species assemblages as high plant diversity supports microbial performance and soil processes, and the expected patterns would be essentially similar in both forests. Allium ursinum L. and Dentaria enneaphyllos L. represented herb species forming mono-dominant patches in beech forest, while Aegopodium podagraria L. and Ficaria verna Huds. represented herb species forming mono-dominant patches in riparian forest. Our hypotheses were only partly supported by the data. We found that herb plant species affected soil microbial communities and processes, particularly in the riparian forest, but they generally did not influence soil physicochemical properties. In the beech forest, herbaceous plants increased saprotrophic fungi biomass, fungi/bacteria ratio, and arylsulfatase activity, with the highest values under D. enneaphyllos. In the riparian forest, a number of microbial parameters, namely bacteria, G+ bacteria, and saprotrophic fungi biomass, fungi/bacteria ratio, and soil respiration exhibited the lowest values in bare soil and the highest values in soil under A. podagraria. Contrary to expectations, soils under multi-species assemblages were characterized by intermediate values of microbial parameters. Concluding, herbaceous plant species largely supported soil microbial communities in deciduous temperate forests but did not affect soil chemical properties. The potential reasons for the positive influence of herb plants on soil microbes (litterfall, rhizodeposition) require further investigation.

Effect of Vancomycin on Cellular Fatty Acid Profiles of Vancomycin-Susceptible and Nonsusceptible Staphylococcus aureus.

Vancomycin is widely used for treatment of infection caused by methicillin-resistant Staphylococcus aureus (MRSA) leading to an increasing appearance of low-level vancomycin-resistant isolates called heterogeneous vancomycin-intermediate S. aureus (hVISA). The mechanism of vancomycin tolerance in hVISA is still unclear. This study aimed to investigate the fatty acid compositions of S. aureus isolates under the stress environment with vancomycin. The different responses of hVISA and vancomycin-susceptible S. aureus (VSSA) may lead to more understanding the mechanism. The bacterial lipid profiles were tested three times from three extractions of each isolate cultured on tryptic soy agar (TSA) and TSA with vancomycin. Of the 30 MRSA isolates studied, 13, 12, and 5 isolates were VSSA, hVISA, and VISA, respectively. The analysis of bacterial lipid profiles showed that under vancomycin stress, there was a reduction of straight chain fatty acids (SCFAs) in VSSA isolates but an increase in branched chain fatty acids (BCFAs). In contrast, the hVISA group exhibited an increase only in the BCFAs but not in SCFAs. Of interest, vancomycin had no effect on either BCFAs or SCFAs of the VISA cells. This study provided information of bacterial adaptation during stress with vancomycin that may be helpful to overcome the resistant bacteria.

Hexanol biosynthesis from syngas by Clostridium carboxidivorans P7 - product toxicity, temperature dependence and in situ extraction.

Clostridium carboxidivorans converts syngas into industrial alcohols like hexanol, but titers may be limited by product toxicity. Investigation of IC50 at 30 °C (17.5 mM) and 37 °C (11.8 mM) revealed increased hexanol tolerance at lower temperatures. To avoid product toxicity, oleyl alcohol was added as an extraction solvent, increasing hexanol production nearly 2.5-fold to 23.9 mM (2.4 g/L) at 30 °C. This titer exceeds the concentration that is acutely toxic in the absence of a solvent, confirming the hypothesis that current hexanol production is limited by product toxicity. The solvent however had no positive effect at 37 °C. Furthermore, C. carboxidivorans cell membranes adapted to the higher temperature by incorporating more saturated fatty acids, but surprisingly not to hexanol. Corn oil and sunflower seed oil were tested as alternative, inexpensive extraction solvents. Hexanol titers were similar with all solvents, but oleyl alcohol achieved the highest extraction efficiency.

Bioremediation of ossein effluents using the filamentous marine cyanobacterium Cylindrospermum stagnale.

Wastewater containg proteinaceous ossein effluents are problematic to be treated. We studied the possibility to treat ossein effluents with the marine cyanobacterium strain Cylindrospermum stagnale. After optimizing the culture conditions of the bacterium, three different types of ossein effluents were tested: dicalcium phosphate (DCP), high total dissolved solids (HTDS) and low total dissolved (LTDS). The effluents were diluted with sea water at the following ratios 1:1, 2:1 and 3:2. The optimum operating conditions were at 3000 lux light intensity and 37 °C temperature. The highest degradation of ossein effluens by C. stagnale was attained for a dilution ratio of 1:1. However, less diluted ossein effluents reduced the growth of C. stagnale drastically. The degradation was shown by measuring the chlorophyll a content and the dry weight of bacterial cells during a seven-day incubation period degradation. Fourier Transform Infrared Spectroscopy (FT-IR) analysis verified the degradation showing the presence of the degradation products of ossein (i.e. calcium carbonate and calcite) in the culture medium. Lipid composition in fatty acids appeared to be suitable for biofuel production. The results showed that the marine cyanobacterium C. stagnale can be used to treat ossein effluents, and at the same time, to produce biofuel in a sustainable way.

Aureimonas mangrovi sp. nov., a marine alphaproteobacterium isolated from mangrove sediment in Thailand.

Two bacterial strains, designated as 1-4-3T and 1-4-4, were isolated from a mangrove sediment cultured with coastal seawater. The cells were Gram-stain-negative, motile, short, rod-shaped bacteria with flagella. Growth occurred at 4-37 °C, pH 7.0-9.0, and 0-7% NaCl. The predominant fatty acids of the novel strains were C18 : 1 ω7c, C19 : 0 cyclo ω8c, C18 : 0, and C16 : 0. A phylogenetic analysis based on 16S rRNA gene sequences and whole genome phylogeny analysis based on distance matrix revealed an affiliation between the two strains and the genus Aureimonas, with closest sequence similarity to A. populi 4M3-2T (96.41 and 96.64% similarity, respectively) and A. glaciistagni (96.01 and 96.23% similarity, respectively). The DNA G+C content of strain 1-4-3T was 66.80 mol%. Strain 1-4-3T displayed low DNA-DNA relatedness to A. populi 4M3-2T, with an average nucleotide identity value of 77.47 % and digital DNA-DNA hybridization value of 22.83 %. Genotypic, chemotaxonomic, and phenotypic data indicate that strains 1-4-3T and 1-4-4 represent a novel species of the genus Aureimonas, for which we propose the name Aureimonas mangrovi sp. nov. The type strain is 1-4-3T (=LMG 31693T=CGMCC 1.18507T).

Fatty Acid Composition by Total Acyl Lipid Collision-Induced Dissociation Time-of-Flight (TAL-CID-TOF) Mass Spectrometry.

Total acyl lipid collision-induced dissociation time-of-flight (TAL-CID-TOF) mass spectrometry uses a quadrupole time-of-flight (QTOF) mass spectrometer to rapidly provide a comprehensive fatty acid composition of a biological lipid extract. Samples are infused into a QTOF instrument, operated in negative mode, and the quadrupole is used to transfer all, or a wide mass range of, precursor ions to the collision cell for fragmentation. Time-of-flight-acquired mass spectra provide mass accuracy and resolution sufficient for chemical formula determination of fatty acids in the complex mixture. Considering the limited number of reasonable CHO variants in fatty acids, one can discern acyl anions with the same nominal mass but different chemical formulas. An online application, LipidomeDB Data Calculation Environment, is employed to process the mass spectral output data and match identified fragments to target fragments at a resolution specified by the user. TAL-CID-TOF methodology is a useful discovery or screening tool to identify and compare fatty acid profiles of biological samples.