Background: Due to the high prevalence and complex etiology, bovine mastitis (BM) is one of the most important diseases to compromise dairy cow health and milk quality. The shift in milk compositions has been widely investigated during mastitis, but recent studies suggested that gastrointestinal microorganism also has a crucial effect on the inflammation of other peripheral tissues and organs, including the mammary gland. However, research focused on the variation of rumen inner-environment during mastitis is still limited. Therefore, the ruminal microbial profiles, metabolites, and milk compositions in cows with different udder health conditions were compared in the present study. Furthermore, the correlations between udder health status and ruminal conditions were investigated. Based on the somatic cell counts (SCC), California mastitis test (CMT) parameters and clinical symptoms of mastitis, 60 lactating Holstein dairy cows with similar body conditions (excepted for the udder health condition) were randomly divided into 3 groups (n = 20 per group) including the healthy (H) group, the subclinical mastitis (SM) group and the clinical mastitis (CM) group. Lactation performance and rumen fermentation parameters were recorded. And rumen microbiota and metabolites were also analyzed via 16S rRNA amplicon sequencing and untargeted metabolomics, respectively. Results: As the degree of mastitis increased, rumen lactic acid (LA) (P < 0.01), acetate, propionate, butyrate, valerate (P < 0.001), and total volatile fatty acids (TVFAs) (P < 0.01) concentrations were significantly decreased. In the rumen of CM cows, the significantly increased bacteria related to intestinal and oral inflammation, such as Lachnospiraceae (FDR-adjusted P = 0.039), Moraxella (FDR-adjusted P = 0.011) and Neisseriaceae (FDR-adjusted P = 0.036), etc., were accompanied by a significant increase in 12-oxo-20-dihydroxy-leukotriene B4 (FDR-adjusted P = 5.97 × 10^− 9) and 10beta-hydroxy-6beta-isobutyrylfuranoeremophilane (FDR-adjusted P = 3.88 × 10^− 10). Meanwhile, in the rumen of SM cows, the Ruminiclostridium_9 (FDR-adjusted P = 0.042) and Enterorhabdus (FDR-adjusted P = 0.043) were increased along with increasing methenamine (FDR-adjusted P = 6.95 × 10^− 6), 5-hydroxymethyl-2-furancarboxaldehyde (5-HMF) (FDR-adjusted P = 2.02 × 10^− 6) and 6-methoxymellein (FDR-adjusted P = 2.57 × 10^− 5). The short-chain fatty acids (SCFAs)-producing bacteria and probiotics in rumen, including Prevoterotoella_1 (FDR-adjusted P = 0.045) and Bifidobacterium (FDR-adjusted P = 0.035), etc., were significantly reduced, with decreasing 2-phenylbutyric acid (2-PBA) (FDR-adjusted P = 4.37 × 10^− 6). Conclusion: The results indicated that there was a significant shift in the ruminal microflora and metabolites associated with inflammation and immune responses during CM. Moreover, in the rumen of cows affected by SM, the relative abundance of several opportunistic pathogens and the level of metabolites which could produce antibacterial compounds or had a competitive inhibitory effect were all increased.

The metabolic activity in plants or fruits is associated with volatile organic compounds (VOCs), which can help identify the different diseases. P-ethylphenol has been demonstrated as one of the most important VOCs released by the Phytophthora cactorum (P. cactorum) infected strawberries. In this study, a bioelectronic nose based on a gas biosensor array and signal processing model was developed for the noninvasive diagnostics of the P. cactorum infected strawberries, which could overcome the limitations of the traditional spectral analysis methods. The gas biosensor array was fabricated using the single-wall carbon nanotubes (SWNTs) immobilized on the surface of field-effect transistor, and then non-covalently functionalized with different single-strand DNAs (ssDNA) through π–π interaction. The characteristics of ssDNA-SWNTs were investigated using scanning electron microscope, atomic force microscopy, Raman, UV spectroscopy, and electrical measurements, indicating that ssDNA-SWNTs revealed excellent stability and repeatability. By comparing the responses of different ssDNA-SWNTs, the sensitivity to P-ethylphenol was significantly higher for the s6DNA-SWNTs than other ssDNA-SWNTs, in which the limit of detection reached 0.13% saturated vapor of P-ethylphenol. However, s6DNA-SWNTs can still be interfered with by other VOCs emitted by the strawberries in the view of poor selectivity. The bioelectronic nose took advantage of the different sensitivities of different gas biosensors to different VOCs. To improve measure precision, all ssDNA-SWNTs as a gas biosensor array were applied to monitor the different VOCs released by the strawberries, and the detecting data were processed by neural network fitting (NNF) and Gaussian process regression (GPR) with high accuracy.

Cadmium (Cd) and lead (Pb) in decorative materials threaten human health. To determine the content of Cd(II) and Pb(II), a disposable screen-printed electrode (DSPE) electrically modified with reduced graphene oxide (rGO) and L-cysteine (LC) was fabricated, which was further electroplated with bismuth film (Bi/LC-rGO/DSPE) in situ. The electrochemical properties of this electrode were studied using cyclic voltammetry, electrochemical impedance spectroscopy, linear sweep voltammetry and differential pulse voltammetry. The results indicated that the Bi/LC-rGO/DSPE had excellent sensitivity, selectivity and stability with low cost and easy production. After optimizing the detection parameters, the linear range of the Bi/LC-rGO/DSPE was from 1.0 to 30.0 μg/L for Cd(II) and Pb(II), and the detection limits were 0.10 μg/L for Cd(II) and 0.08 μg/L for Pb(II). Finally, the Bi/LC-rGO/DSPE was applied to determine the concentrations of Cd(II) and Pb(II) in different decorative materials where the recoveries were in the range from 95.86% to 106.64%.

Background: Kelp powder, which was rich in novel oligosaccharides and iodine might be utilized by the rumen microbiome, promoted the ruminal fermentation and finally enhanced the lactation performance of dairy cows. Therefore, the purpose of this study was to investigate the effects of kelp powder partially replacing dietary forage on rumen fermentation and lactation performance of dairy cows. (2) Methods: In the present study, 20 Chinese Holstein dairy cows were randomly divided into two treatments, a control diet (CON) and a kelp powder replacing diet (Kelp) for a 35-d long trial. Dry matter intake (DMI), milk production, milk quality, ruminal fermentable parameters, and rumen microbiota were measured to investigate the effects of kelp powder feeding on dairy cows. (3) Results: On the lactation performance, kelp significantly increased milk iodine content and effectively enhanced milk production and milk fat content. On the fermentable aspects, kelp significantly raised TVFA while reducing the ammonia-N content. On the rumen microbial aspect, kelp feeding significantly promoted the proliferation of Firmicutes and Proteobacteria while suppressing Bacteroidetes. (4) Conclusion: kelp powder as an ingredient of feedstuff might promote the rumen fermentation ability and effectively increase milk fat and iodine content, and consequently improve the milk nutritional value.

As the co-enzyme of pyruvate formate-lyase under ruminal anaerobic condition, thiamine plays a critical role in carbohydrate metabolism in dairy cows. The objective of this study was to investigate the impacts of thiamine supplementation on ruminal carbohydrate-active enzymes. Twelve Holstein dairy cows were randomly assigned into three dietary treatments: control diet (CON; 20% starch, dry matter (DM) basis), high-concentrate diet (HC; 33.2% starch, DM basis) and a high-concentrate diet supplemented with 180 mg thiamine/kg DM (HCT; 33.2% starch, DM basis). Dry matter intake and milk production were recorded for 21 days. Rumen fluid samples were collected, and ruminal pH and volatile fatty acids (VFAs) were measured. The metagenome sequencing technique was used to detect the genes in ruminal microorganisms and identify putative carbohydrate-active enzymes. The total abundances of carbohydrate-active enzymes and fiber-degrading enzymes were both reduced by HC with no effect on starch-degrading enzymes compared with CON. However, the fiber-degrading enzymes and starch-degrading enzymes were both increased after thiamine supplementation. These results indicated that 180 mg thiamine /kg DM might effectively improve rumen carbohydrate metabolism through increasing the abundance of ruminal carbohydrate-active enzymes and consequently balanced the rumen volatile fatty acids and rumen pH, providing a practical strategy in preventing subacute ruminal acidosis in cows offered HC.

Hypocalcemia is caused by a sharp decline in blood calcium concentration after dairy cow calving, which can lead to various diseases or even death. It is necessary to develop an inexpensive, easy-to-operate, reliable sensor to diagnose hypocalcemia. The cellulose-paper-based microfluidic field-effect biosensor is promising for point-of-care, but it has poor mechanical strength and a short service life after exposure to an aqueous solution. Octadecyltrichlorosilane (OTS), as a popular organosilane derivative, can improve the hydrophobicity of cellulose paper to overcome the shortage of cellulose paper. In this work, OTS was used to produce the superhydrophobic cellulose paper that enhances the mechanical strength and short service life of MFB, and a microfluidic field-effect biosensor (MFB) with semiconducting single-walled carbon nanotubes (SWNTs) and DNAzyme was then developed for the Ca²⁺ determination. Pyrene carboxylic acid (PCA) attached to SWNTs through a non-covalent π-π stacking interaction provided a carboxyl group that can bond with an amino group of DNAzyme. Two DNAzymes with different sensitivities were designed by changing the sequence length and cleavage site, which were functionalized with SPFET/SWNTs-PCA to form Dual-MFB, decreasing the interference of impurities in cow blood. After optimizing the detecting parameters, Dual-MFB could determine the Ca²⁺ concentration in the range of 25 μM to 5 mM, with a detection limit of 10.7 μM. The proposed Dual-MFB was applied to measure Ca²⁺ concentration in cow blood, which provided a new method to diagnose hypocalcemia after dairy cow calving.