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Short-chain fatty acids (SCFA) and lactate in ruminal fluid are products resulting from the microbial fermentation of substrates and can be used to reflect the composition and activity of the ruminal microbiome. Determination of SCFA and D-/L-lactate in ruminal fluid currently requires two separate protocols, which is time-consuming and costly. In this study, we have optimised and validated a simple and unified 3-nitrophenylhydrazine (3-NPH) derivatisation protocol and a 20 min chiral-LC-MS method for the simultaneous quantification of all SCFA and D- and L-lactate in ruminal fluid. This method, which requires no sample pretreatment or purification shows adequate sensitivity (limit of detection (LOD): 0.01 µg/mL), satisfactory accuracy (recovery: 88–103%), and excellent reproducibility (relative standard deviation (RSD) for repeated analyses < 3% for most analytes). The application of this method to a cohort of 24 animals allowed us to reveal a large inter-cow variation in ruminal SCFA and lactate level, the concentration range for each species, the widespread correlation between different SCFA, and the strong correlation between D- and L-lactate. [ABSTRACT FROM AUTHOR]

An accurate dry matter intake (DMI) measurement method is of great importance for determining the performance of dairy cows. While n-alkane technique is commonly used to estimate the DMI of ruminants, the protocols used to measure alkanes are complex, time consuming and low throughput. Herein we present a simplified and miniaturised protocol that combines a smaller sample matrix (60 mg of faeces or herbage), a shorter (3 h) saponification step and a direct purification of the crude extract by filtration through silica powder or mixing with silica powder prior to GC-FID analysis. In addition to a much higher throughput compared with the existing methods, by skipping the commercial SPE columns and reducing the organic solvent volume, our miniaturised protocol should significantly reduce the sample processing cost and is thus suitable for alkane determination for large-scale experiments. [ABSTRACT FROM AUTHOR]

Simple Summary: Batocera horsfieldi (Hope) is an important wood-boring pest in China. This pest primarily infests tree trunks by feeding on the woody tissue, creating a network of interconnected tunnels within the trunk. These tunnels become blocked with insect feces and wood debris, causing damage, decay, and even the death of the host plant's tissues. In this study, single-molecule real-time sequencing (SMRT) and Illumina RNA-seq technologies were employed to conduct full-length transcriptome sequencing of male and female adults of B. horsfieldi. A total of 20,356,793 subreads (38.26 G, clean reads) were generated, including 432,091 circular consensus sequences and 395,851 full-length non-chimera reads. Clustering and redundancy removal of the full-length non-chimera reads resulted in 39,912 consensus reads. Additionally, functional annotation was performed on a total of 84,650 transcripts in seven different databases. This study provides an important foundation for future exploration of gene regulation in the interaction between B. horsfieldi and host plants using RNA interference (RNAi), and it offers a scientific basis for the prevention and control of B. horsfieldi. Batocera horsfieldi (Hope) (Coleoptera: Cerambycidae) is an important forest pest in China that mainly infests timber and economic forests. This pest primarily causes plant tissue to necrotize, rot, and eventually die by feeding on the woody parts of tree trunks. To gain a deeper understanding of the genetic mechanism of B. horsfieldi, this study employed single-molecule real-time sequencing (SMRT) and Illumina RNA-seq technologies to conduct full-length transcriptome sequencing of the insect. Total RNA extracted from male and female adults was mixed and subjected to SMRT sequencing, generating a complete transcriptome. Transcriptome analysis, prediction of long non-coding RNA (lncRNA), coding sequences (CDs), analysis of simple sequence repeats (SSR), prediction of transcription factors, and functional annotation of transcripts were performed in this study. The collective 20,356,793 subreads (38.26 G, clean reads) were generated, including 432,091 circular consensus sequences and 395,851 full-length non-chimera reads. The full-length non-chimera reads (FLNC) were clustered and redundancies were removed, resulting in 39,912 consensus reads. SSR and ANGEL software v3.0 were used for predicting SSR and CDs. In addition, four tools were used for annotating 6058 lncRNAs, identifying 636 transcription factors. Furthermore, a total of 84,650 transcripts were functionally annotated in seven different databases. This is the first time that the full-length transcriptome of B. horsfieldi has been obtained using SMRT sequencing. This provides an important foundation for investigating the gene regulation underlying the interaction between B. horsfieldi and its host plants through gene editing in the future and provides a scientific basis for the prevention and control of B. horsfieldi. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Regioisomers (or positional isomers) of triacylglycerols (TAGs) of milk are known to show differential outcome in relation to human absorption. Quantitation of TAG regioisomers remains a big challenge due to the lack of facile chromatographic separation technique. The feasibility of using fragment ion intensity ratio to determine the ratio of co-eluting AAB/ABA-type regioisomer pairs was confirmed in this study. The ability of C30 stationary phase in resolving interfering TAG isomers was demonstrated for the first time. This allowed us to reveal the complexity of using fragment ion intensity to quantify 1,2-olein-3-palmitin (OOP), 1,3-olein-2-palmitin (OPO), 1,2-olein-3-stearin (OOS), and 1,3-olein-2-stearin (OSO) regioisomers in milk samples. A novel algorithm was proposed to consider the contribution of OPO/OOP and OSO/OOS double bond (DB)-isomers and to eliminate the interference of isobaric ions from other isomers, an aspect overlooked in previous studies. This liquid chromatography-mass spectrometry method that requires no pre-fractioning and a moderate chromatographic separation time of 36 min is simple and, thus, suitable for screening a large number of samples for genetic analysis of this trait. Preliminary results using a small cohort of animals showed that OPO/OOP ratio differs significantly between Jersey and Holstein cows, and a large variation was also observed across individual Holstein cows. [ABSTRACT FROM AUTHOR]

Ensuring the large signal stability of the DC microgrid is the premise of the safe operation of the DC microgrid, but the research on the large-signal stability of microgrids with multiple droop control micro-sources is still scarce. In this paper, a DC microgrid system model with multiple droop control micro-sources was established by appropriate simplification. Addressing the problem that most stability research methods cannot be quantitatively analyzed, the mixed potential function method was used to analyze the large signal stability of the system. However, the criterion obtained by the conventional mixed potential function method is complicated and contains multiple time-varying parameters, which is not convenient for analysis. Therefore, the simple form of the criterion was obtained through simplification and the analysis proved the rationality of the simplification. On this basis, a nonlinear droop control method was proposed to improve the anti-interference ability of the system. Finally, the accuracy of the large signal stability criterion and the effectiveness of nonlinear droop control on the system's large signal stability were verified by simulation. [ABSTRACT FROM AUTHOR]