Microbiota Characterization of the Cow Mammary Gland Microenvironment and Its Association with Somatic Cell Count.

Subclinical mastitis is a common disease that threatens the welfare and health of dairy cows and causes huge economic losses. Somatic cell count (SCC) is the most suitable indirect index used to evaluate the degree of mastitis. To explore the relationship between SCC, diversity in the microbiome, and subclinical mastitis, we performed next-generation sequencing of the 16S rRNA gene of cow's milk with different SCC ranges. The data obtained showed that the microbiota was rich and coordinated with SCC below 2 × 105. SCC above 2 × 105 showed a decrease in the diversity of microbial genera. When SCC was below 2 × 105, the phylum Actinobacteriota accounted for the most. When SCC was between 2 × 105 and 5 × 105, Firmicutes accounted for the most, and when SCC exceeded 5 × 105, Firmicutes and Proteobacteria accounted for the most. Pathogenic genera such as Streptococcus spp. were absent, while SCC above 2 × 105 showed a decrease in the diversity of microbial genera. SCC was positively correlated with the percentage of Romboutsia, Turicibacter, and Paeniclostridium and negatively correlated with the percentage of Staphylococcus, Psychrobacter, Aerococcus, and Streptococcus. Romboutsia decreased 6.19 times after the SCC exceeded 2 × 105; the SCC increased exponentially from 2 × 105 to 5 × 105 and above 1 × 106 in Psychrobacter. Analysis of the microbiota of the different SCC ranges suggests that the development of mastitis may not only be a primary infection but may also be the result of dysbiosis in the mammary gland.

Bactericidal Mechanisms of Chlorine Dioxide against Beta-Hemolytic Streptococcus CMCC 32210.

Chlorine dioxide is a globally recognized green and efficient disinfectant. This study aims to investigate the bactericidal mechanism of chlorine dioxide using beta-hemolytic Streptococcus (BHS) CMCC 32210 as a representative strain. BHS was exposed to chlorine dioxide, the minimum bactericidal concentration (MBC) values of chlorine dioxide against BHS were determined by the checkerboard method in preparation for subsequent tests. Cell morphology was observed using electron microscopy. Protein content leakage, adenosine triphosphatase (ATPase) activity, and lipid peroxidation were determined by kits, and DNA damage was determined using agar gel electrophoresis. The concentration of chlorine dioxide during disinfection showed a linear relationship with the concentration of BHS. Scanning electron microscopy (SEM) results showed that chlorine dioxide caused significant damage to the cell walls of BHS at a concentration of 50 mg/L, but had no significant effect on Streptococcus exposed to different exposure times. Furthermore, the extracellular protein concentration increased with increasing chlorine dioxide concentration, while the total protein content remained unchanged. The activities of Na+/K+-ATPase and Ca2+/Mg2+-ATPase decreased with increasing chlorine dioxide concentration. Chlorine dioxide treatment led to significant lipid peroxidation and DNA degradation in BHS. Leakage of intracellular components indicated that chlorine dioxide damaged the cell membrane of BHS. Chlorine dioxide exposure resulted in oxidative damage to lipids and proteins, which negatively impacted the cell wall and membrane of Streptococcus. This caused increased permeability and inactivation of key enzymes (Na+/K+-ATPase and Ca2+/Mg2+-ATPase) involved in respiratory metabolism, ultimately leading to DNA degradation and bacterial death due to either content leakage or metabolic failure.

Integration of Network Pharmacology and Molecular Docking to Analyse the Mechanism of Action of Oregano Essential Oil in the Treatment of Bovine Mastitis.

The active components, potential targets, and mechanisms of action of oregano essential oil in the treatment of bovine mastitis disease were investigated using network pharmacology and molecular docking approaches. The TCMSP and literature databases were examined for the main compounds in oregano essential oil. Afterward, the physical, chemical, and bioavailability characteristics of the components were evaluated. The PubChem, BATMAN, PharmMapper, and Uniprot databases were utilized to predict the target genes of the major components of oregano essential oil. Via the databases of DrugBank, OMIM, GeneCards, TTD, and DisGenet, the disease targets of bovine mastitis were discovered. We analyzed common targets and built protein-protein interaction (PPI) networks using the STRING database. Key genes were analyzed, obtained, and compound-target-pathway-disease visualization networks were created using Cytoscape. For the GO function and KEGG pathway enrichment analysis, the DAVID database was utilized. Molecular docking via Autodock Tools was utilized to evaluate the reliability of the interactions between oregano essential oil and hub targets. Thymol, carvacrol, and p-cymene are the three major components found in oregano essential oil. The potential targets (TNF, TLR4, ALB, IL-1ß, TLR2, IL-6, IFNG, and MyD88) were screened according to the visual network. The enrichment analysis suggested that the major signaling pathways in network pharmacology may include PI3K-Akt, MAPK, IL-17, and NF-κ B. Molecular docking analysis shows that thymol had good docking activity with TNF, IL-6, and MyD88, carvacrol had good docking activity with TNF, and p-cymene had good docking activity with ALB. This study clarified the mechanism of action of oregano essential oil in the treatment of bovine mastitis, thus providing data supporting the potential for the use of oregano essential oil in the development of new therapeutics for bovine mastitis.

[Treatment of degenerative medial meniscus injury of knee joint by arthroscopy combined with small needle knife to release superficial medial collateral ligament of knee joint].

OBJECTIVE: To explore the clinical effect of arthroscopic combined with small needle knife in the treatment of degenerative medial meniscus (MM) injury of knee joint by releasing the superficial layer of medial collateral ligament (SMCL). METHODS: From February 2016 to November 2018, 56 patients (56 knees) with limited pain, strangulation and flexion in medial knee joint space were selected. X-ray Kellgren-Lawrence grading was I-II. MRI showed medial meniscus injury(III degree) of knee joint. There were 30 males(30 knees) and 26 females(26 knees). Arthroscopic MM plasty and small needle knife were used to release SMCL. The Lysholm knee score was used to evaluate the effect of operation. RESULTS: All 56 patients were followed up, and the duration ranged from 3 to 24 months, with an average of 10 months. According to the Lysholm knee score standard, the final follow-up was compared with that of before operation. The results showed that the preoperative knee score was 37.24±1.32, the latest follow-up knee score was 85.72±5.28, the knee score was higher than that before the operation(P<0.05). CONCLUSIONS: Arthroscopy combined with small needle knife release of superficial medial collateral ligament in the treatment of degenerative medial knee meniscus injury can effectively improve the mechanical balance of the knee joint, improve Lysholm knee score in patients with knee meniscus injury, and promote the recovery of knee joint function, which has clinical value.

Extraction and Analysis of Available Boron Isotopes in Soil Using Multicollector Inductively Coupled Plasma Mass Spectrometry.

As a result of the important roles of boron (B) in the growth of plants, the uptake of B by plants is dependent upon the existing form and content of available B in soil, which can bring about the local cycle of B isotope equilibrium. A method using water-heating extraction combined with three-step ion-exchange chromatography was developed for the extraction and isotopic analysis of available B in soil. The extraction efficiency and fractionation of B isotopic composition in the procedure were investigated. The results showed that, in the upper layers of soils, the change of δ11B values was opposite that of the mass concentration and a similar variation between δ11B and content occurred in the lower layers. The isotope of available B in soil can create a featured isotopic signature to further understand the geochemical details related to the soil properties and molecular mechanism of B uptake in plants.