Optimised Method for Short-Chain Fatty Acid Profiling of Bovine Milk and Serum

Short-chain fatty acids (SCFA, C2-C5) in milk and serum are derived from rumen bacterial fermentation and, thus, have the potential to be used as biomarkers for the health status of dairy cows. Currently, there is no comprehensive and validated method that can be used to analyse all SCFAs in both bovine serum and milk. This paper reports an optimised protocol, combining 3-nitrophenylhydrazine (3-NPH) derivatisation and liquid chromatography-mass spectrometry (LC-MS) analysis for quantification of SCFA and β-hydroxybutyric acid (BHBA) in both bovine milk and bovine serum. This method is sensitive (limit of detection (LOD) ≤ 0.1 µmol/L of bovine milk and serum), accurate (recovery 84–115% for most analytes) and reproducible (relative standard deviation (RSD) for repeated analyses below 7% for most measurements) with a short sample preparation step. The application of this method to samples collected from a small cohort of animals allowed us to reveal a large variation in SCFA concentration between serum and milk and across different animals as well as the strong correlation of some SCFAs between milk and serum samples.

Fatty acid profiling in gobhi sarson (Brassica napus)

An experiment was carried out during 2017-2018 to estimate fatty acids and the oil content (OC) in fifteen Brassica napus genotypes. The quality parameters of oil include fatty acids (FA) and the oil content (OC), important trait differed significantly (p?0.05) amongst the Brassica species genotype. Among the genotypes, significant differences were noted for the fatty acids and the oil content (OC). In Brassica napus seeds oil content varies in between the range of 37.45–41.86% respectively. The saturated fatty acid (SFA) includes the Palmitic acid (PA) varied in between the range of 2.68–4.43% and oleic acid (OA) content results lied between 8.88-56.18% respectively. In linoleic acid (LA) and linolenic acid (LNA), presence of significant differences (p?0.05) was there. The content of linoleic acid (LA) lies in the range between 12.97- 17.98% respectively and linolenic acid (LNA) content varied from 13.41-23.42% respectively. The stearic acid (SA) content varied from1.20-1.66 respectively. Erucic acid, another essential trait, significant differences were noted amongst the Brassica species genotypes i.e. 12.96-48.80%. The minimum erucic acid (EA) content was noted in GSL-1 genotype and the genotypes namely RSPN-28 and CNH-13-2, EC552608, GSC-6 have also low EA content and the rest of the genotypes namely, RSPN-29, DGS-1, RSPN-25, CNH-11-7, CNH-11-13, RL-1359, HNS-1101, GSC-101, CNH-11-2, HNS-1102 have high erucic content. In fatty acids (FA) content, significant differences were observed in rapeseed-mustard. Desirable cultivars with higher yield and oil content are the chief objective of this concerned study to be further employed in the breeding program.

Automated Trimethyl Sulfonium Hydroxide Derivatization Method for High-Throughput Fatty Acid Profiling by Gas Chromatography–Mass Spectrometry

Fatty acid profiling on gas chromatography–mass spectrometry (GC–MS) platforms is typically performed offline by manually derivatizing and analyzing small batches of samples. A GC–MS system with a fully integrated robotic autosampler can significantly improve sample handling, standardize data collection, and reduce the total hands-on time required for sample analysis. In this study, we report an optimized high-throughput GC–MS-based methodology that utilizes trimethyl sulfonium hydroxide (TMSH) as a derivatization reagent to convert fatty acids into fatty acid methyl esters. An automated online derivatization method was developed, in which the robotic autosampler derivatizes each sample individually and injects it into the GC–MS system in a high-throughput manner. This study investigated the robustness of automated TMSH derivatization by comparing fatty acid standards and lipid extracts, derivatized manually in batches and online automatically from four biological matrices. Automated derivatization improved reproducibility in 19 of 33 fatty acid standards, with nearly half of the 33 confirmed fatty acids in biological samples demonstrating improved reproducibility when compared to manually derivatized samples. In summary, we show that the online TMSH-based derivatization methodology is ideal for high-throughput fatty acid analysis, allowing rapid and efficient fatty acid profiling, with reduced sample handling, faster data acquisition, and, ultimately, improved data reproducibility.

Comparative Fatty Acid Profiling of Edible Fishes in Kuala Terengganu, Malaysia

The aim of this study was to compare the relative nutritional benefit of edible Malaysian fishes from the coast of Terengganu in Malaysia, as well as to perform a taxonomical characterization and metal assessment. Discrimination between species was carried out by a morphological and molecular approach by evaluating the total concentrations of metals by ICP-MS analyses and the fatty acids (FA) composition using the GC–MS approach on the fish fillet tissues. The taxonomical studies detected fishes of 11 families and 13 species. The heavy metal assessment showed that all detected elements did not exceed the regulatory limit stated by Malaysian Food Regulations. The proportion of saturated fatty acids (SFA) ranged from 33 to 58.34%, followed by the polyunsaturated fatty acids (PUFA) values from 24 to 51.8%, and the lowest proportion was of monounsaturated fatty acids (MUFA), ranging from 12.7 to 35.9%. The ω-3/ω-6 PUFA and PUFA/SFA ratios were determined in the range 1.1 to 7.4 and 0.35 to 1.6, respectively. The C20:5 ω-3 and C22:6 ω-3 acids were detected at levels comparable to those found in the corresponding species from similar tropical marine ecosystems. The high FA values can be useful biochemical tools for comparing the relative nutritional benefits of these biodiverse and non-toxic edible Malaysian fishes.

Fatty Acid Profiling of Moina sp. Preserved in Cryoprotective Agents at Low Temperature

Highlight ResearchSaturated fatty acid (SFA), polyunsaturated fatty acid (PUFA), ∑ ω6 and ∑ ω3 of Moina sp. preserved with 5, 10 and 20% GLY decreased with prolong storage.FA of Moina sp. preserved with 5, 10 and 20% EG showed a significant reduced only after M3.Monounsaturated fatty acid (MUFA) and PUFA of Moina sp. preserved with 5, 10 and 20% DMSO increased significantly (p<0.05) after M3.Moina sp. preserved in 5% DMSO maintain the docosaheaenoic acid (DHA) level, which could be potential method for long-term preservation.AbstractMoina sp. is an important feed for larval and post-larval rearing of aquaculture species. Preservation of Moina sp. using various preservation agents and techniques is known to be less time-consuming procedure in maintaining large number of feed in hatchery management. Hence, this study was carried out to determine the changes of fatty acid (FA) composition of Moina sp. preserved at 5, 10 and 20% in glycerol (GLY), ethylene glycol (EG) and dimethyl sulfoxide (DMSO) for 1 to 3 months (M1-M3). Moina sp. without cryoprotectant agents stored at -40°C as controls. Saturated fatty acid (SFA), polyunsaturated fatty acid (PUFA), ∑ ω6 and ∑ ω3 of Moina sp. preserved with 5, 10 and 20% GLY decreased with prolonging storage. FA of Moina sp. preserved with 5, 10 and 20% EG showed a significant reduction only after M3. Monounsaturated fatty acid (MUFA) and PUFA of Moina sp. preserved with 5, 10 and 20% DMSO increased significantly (p<0.05) after M3. Moina sp. preserved in 5% DMSO maintain the docosaheaenoic acid (DHA) level, which could be a potential method for long-term preservation.

Shifts in Soil Structure, Biological, and Functional Diversity Under Long-Term Carbon Deprivation

Soil organic matter is composed of a variety of carbon (C) forms. However, not all forms are equally accessible to soil microorganisms. Deprivation of C inputs will cause changes in the physical and microbial community structures of soils; yet the trajectories of such changes are not clear. We assessed microbial communities using phospholipid fatty acid profiling, metabarcoding, CO2 emissions, and functional gene microarrays in a decade-long C deprivation field experiment. We also assessed changes in a range of soil physicochemical properties, including using X-ray Computed Tomography imaging to assess differences in soil structure. Two sets of soils were deprived of C inputs by removing plant inputs for 10 years and 1 year, respectively. We found a reduction in diversity measures, after 10 years of C deprivation, which was unexpected based on previous research. Fungi appeared to be most impacted, likely due to competition for scarce resources after exhausting the available plant material. This suggestion was supported by evidence of bioindicator taxa in non-vegetated soils that may directly compete with or consume fungi. There was also a reduction in copies of most functional genes after 10 years of C deprivation, though gene copies increased for phytase and some genes involved in decomposing recalcitrant C and methanogenesis. Additionally, soils under C deprivation displayed expected reductions in pH, organic C, nitrogen, and biomass as well as reduced mean pore size, especially in larger pores. However, pore connectivity increased after 10 years of C deprivation contrary to expectations. Our results highlight concurrent collapse of soil structure and biodiversity following long-term C deprivation. Overall, this study shows the negative trajectory of continuous C deprivation and loss of organic matter on a wide range of soil quality indicators and microorganisms.

GC-MS Characterization, in vitro Antioxidant, Anti-inflammatory and Potential Antidermatophytic Activity of Citrullus lanatus Seed Oil

Background: Dermatophytes are well-known fungus having the capability to invade keratinized tissues of nails, skin, and hair, of humans and other animals, causing dermatophytosis. Citrullus lanatus (watermelon) seed is a rich source of vitamin E, essential fatty acids, and minerals, bearing various health benefits. Despite the presence of many useful components, the oil is not much explored. Accordingly, the study was planned to investigate the fatty acid profiling, in vitro antioxidant, anti-inflammatory, and potential antidermatophytic activity of Citrullus lanatus seed oil against dermatophytic strains. Methods: The C.lanatus oil was extracted from seeds, and fatty acid profiling was characterized by GC-MS. Further, the oil was subjected to DPPH scavenging activity, and the anti-inflammatory activity was evaluated from Bovine serum albumin assay. The antidermatophytic screening against Microsporum canis, Trichophyton rubrum, and Trichophyton mentagrophytes was carried out by agar well diffusion, broth microdilution assay, and growth kinetics assay. Results: The C. lanatus seed oil exhibited good antioxidant activity close to ascorbic acid, i.e., IC50- 52.22 µg/ml, 35.72 µg/ml, respectively, and the oil exhibited moderate anti-inflammatory activity (315.2 µg/ml) as that of the standard drug diclofenac sodium (174.3µg/ml). The fatty acid profiling indicates the presence of eight main compounds, out of which ç-tocopherol, butylated hydroxytoluene, and hexadecanoic acid are predominantly present. The C.lanatus seed oil showed promising activity against T. rubrum with a lower minimum inhibitory concentration value with respect to positive controls, Fluconazole and Ketoconazole. The oil also had a strong effect on time-dependent kinetic inhibition against T. rubrum. This was the first report of C.lanatus seed oil against any dermatophyte. Conclusion: In conclusion, the study supports the exploitation of C.lanatus seed oil and its main compounds as a source of natural antioxidants, anti-inflammatory, and as a novel agent against dermatophytes for curing superficial fungal infections in the near future.