The somatic cell count (SCC) and total plate count (TPC) are essential quality indicators for raw milk. Traditional detection methods require separate measurements and rely on complex, large-scale instruments or cultivation techniques, which are both time-consuming and laborious. To address these challenges, this study developed a novel method for the simultaneous detection of SCC and TPC in the same raw milk sample using the ATP bioluminescence assay. This method utilizes oxy-ethylated iso-nonyl phenol (Neonol-10) and cetyltrimethylammonium bromide (CTAB) to selectively lyse somatic cells and microorganisms, respectively. This technique is straightforward to operate and can be completed within 2.5 h, with detection ranges of 1 × 10⁴ to 3 × 10⁶ cells/mL for SCC and 1 × 10⁵ to 5 × 10⁷ CFU/mL for TPC. Importantly, this technique meets the requirements of detection standards in China, European Union, Canada, United States, etc. For SCC or TPC in raw milk. Overall, this innovative approach does not rely on expensive equipment or facilities and the stepwise reagent addition procedure can be easily developed into an automated high-throughput system for rapid on-site testing of SCC and TPC in raw milk.
Carbapenemase-producing E. coli is a grave public health concern as the potential emergence of resistant strains and their transmission. Isoorientin belongs to a potential antimicrobial flavonoid compound existing in several plants, while the research on the antimicrobial activity of isoorientin is limited thus far. We evaluated the antimicrobial and antibiofilm effects of isoorientin against biofilm-forming carbapenem non-sensitive Escherichia coli (E. coli) from raw milk of goats, and explored its molecular mechanisms. Isoorientin showed obvious antimicrobial ability with the minimum inhibitory concentration (MIC), and it exhibited synergistic activity with traditional antimicrobials against the carbapenem non-sensitive E. coli. Isoorientin could also significantly inhibit the carbapenem non-sensitive E. coli biofilm formation and destroy the established biofilms, with the percentage of inhibition ranging from 27.8% to 75% at MIC, and the corresponding percentage of eradication ranging from 15.3% to 61.6%, respectively. Confocal laser scanning microscopy (CLSM) observation and scanning electron microscopy (SEM) images indicated that the E. coli biofilm reduced in thickness with increasing concentrations of isoorientin. Dose-dependent decrease in eDNA revealed that isoorientin interacted with the extracellular polymeric substances (EPS) of the biofilm. qRT-PCR assay for the biofilm-forming associated genes further confirmed the above results. Overall, these results concluded that the isoorientin has significant antimicrobial and antibiofilm activity against carbapenem non-sensitive E. coli, and has potential application in prevention of food contamination and spoilage.
Pengjuan Ni,Haichao Dai,Yilin Wang,Yujing Sun,Yan Shi,Jingting Hu,Zhuang Li
In this study, a facile method to sensitively detect melamine and highly improve the peroxidase-like activity of bare gold nanoparticles (Au NPs) at the same time is proposed for the first time. It is interesting to find that the addition of melamine could improve the peroxidase-like activity of Au NPs. By coupling with 3,3′,5,5′-tetramethlybenzidine (TMB)–H 2 O 2 chormogenic reaction, a novel method for colorimetic detection of melamine is developed. The detection limit of this method is as low as 0.2 nM with the help of UV–vis spectroscopy and 0.5 µM by naked-eye observation, both which are far below the US food and Drug Administration estimated melamine safety limit of 20 µM. In addition, the present method is successfully applied for the detection of melamine in raw milk and milk powder. More importantly, the proposed method could also improve the peroxidase-like activity of Au NPs, which may not only provide a new approach to develop effective nanomaterials-based mimetic enzyme , but also irradiative to develop new applications for Au NPs in varieties of cost-effective and simple sensors in medicine, biotechnology and environmental chemistry.
危险品化学品经营许可证（带存储）