Citral is a flavor component that is commonly used in food, beverage and fragrance industries. Cronobacter sakazakii is a food-borne pathogen associated with severe illness and high mortality in neonates and infants. The objective of the present study was to evaluate antimicrobial effect of citral against C. sakazakii strains. The minimum inhibitory concentration (MIC) of citral against C. sakazakii was determined via agar dilution method, then Gompertz models were used to quantitate the effect of citral on microbial growth kinetics. Changes in intracellular pH (pHin), membrane potential, intracellular ATP concentration, and membrane integrity were measured to elucidate the possible antimicrobial mechanism. Cell morphology changes were also examined using a field emission scanning electron microscope. The MICs of citral against C. sakazakii strains ranged from 0.27 to 0.54 mg/mL, and citral resulted in a longer lag phase and lower growth rate of C. sakazakii compared to the control. Citral affected the cell membrane of C. sakazakii, as evidenced by decreased intracellular ATP concentration, reduced pHin, and cell membrane hyperpolarization. Scanning electron microscopy analysis further confirmed that C. sakazakii cell membranes were damaged by citral. These findings suggest that citral exhibits antimicrobial effect against C. sakazakii strains and could be potentially used to control C. sakazakii in foods. However, how it works in food systems where many other components may interfere with its efficacy should be tested in future research before its real application.

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Cronobacter sakazakii is an opportunistic pathogen transmitted by food that affects mainly newborns, infants, and immune-compromised adults. In this study, the antibacterial activity of ferulic acid was tested against C. sakazakii strains. Minimum inhibitory concentration of ferulic acid against C. sakazakii strains was determined using the agar dilution method. Changes in intracellular pH, membrane potential and intracellular ATP concentration were measured to elucidate the possible antibacterial mechanism. Moreover, SYTO 9 nucleic acid staining was used to assess the effect of ferulic acid on bacterial membrane integrity. Cell morphology changes were observed under a field emission scanning electron microscope. The minimum inhibitory concentrations of ferulic acid against C. sakazakii strains ranged from 2.5 to 5.0 mg/mL. Addition of ferulic acid exerted an immediate and sustained inhibition of C. sakazakii proliferation. Ferulic acid affected the membrane integrity of C. sakazakii, as evidenced by intracellular ATP concentration decrease. Moreover, reduction of intracellular pH and cell membrane hyperpolarization were detected in C. sakazakii after exposure to ferulic acid. Reduction of green fluorescence indicated the injury of cell membrane. Electronic microscopy confirmed that cell membrane of C. sakazakii was damaged by ferulic acid. Our results demonstrate that ferulic acid has moderate antimicrobial activity against C. sakazakii. It exerts its antimicrobial action partly through causing cell membrane dysfunction and changes in cellular morphology. Considering its antimicrobial properties, together with its well-known nutritional functions, ferulic acid has potential to be developed as a supplement in infant formula or other foods to control C. sakazakii.

BackgroundThe Lysholm Knee Score (LKS) is widely used and is one of the most effective questionnaires employed to assess knee injuries. Although LKS has been translated into multiple languages, there is no Chinese version even though China has the largest population of patients with knee-joint injuries. The objective of our study was to develop the Chinese version of LKS (C-LKS) and assess its reliability, validity and responsiveness in Chinese patients with anterior cruciate ligament (ACL) injuries.MethodsStudy participants were mainly recruited among patients with ACL injuries scheduled for arthroscopic ACL reconstruction at our hospital. First, we developed the C-LKS in a five-step translation and cross-cultural adaptation procedure. Next, we calculated the Cronbach’s alpha, intraclass correlation coefficient (ICC), Pearson’s correlation coefficient (r), effect size (ES), and standardized response mean (SRM) to evaluate the reliability, validity, and responsiveness of C-LKS respectively.ResultsOverall, 126 patients with ACL injuries successfully completed the questionnaires. Acceptable internal consistency (Cronbach’s alpha = 0.726) as well as excellent test-retest reliability (ICC = 0.935) was found for C-LKS. Good or moderate correlation (r = 0.514–0.837) was determined among C-LKS and International Knee Documentation Committee Subjective Knee Form (IKDC), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), physical subscales of SF-36; C-LKS also had fair or moderate correlation (r = 0.207–0.462) with the other subscales of SF-36, which adequately illustrated that good validity was included in C-LKS. In addition, good responsiveness was also observed in C-LKS (ES = 1.36,SRM = 1.26).ConclusionsWe have shown that our developed C-LKS questionnaire is reliable, valid and responsible for the evaluation of Chinese-speaking patients with ACL injuries and it would be an effective instrument.

Compared with TACE, TACE-MWA leads to better responses for HCC tumors ≤5 cm.