Correlation of PK/PD Indices with Resistance Selection for Cefquinome against Staphylococcus aureus in an In Vitro Model

Cefquinome is a fourth-generation Cephalosporin approved for use in animals exclusively. The objective of this study was to explore the relationship of cefquinome pharmacokinetic/pharmacodynamic (PK/PD) indices with resistance selection of Staphylococcus aureus ATCC25923 in an in vitro model. Six dosing regiments of cefquinome at an interval of 24 h for three consecutive times were simulated, resulting in maximum concentrations (Cmax) from 1/2 MIC to 16 MIC and half-lives (t1/2β) of 3 and 6 h, respectively. The in vitro sensitivity of S. aureus was monitored by bacterial susceptibility and dynamic time-kill curve experiments over the six cefquinome concentrations. The correlation between changes in bacterial susceptibility (MIC72/MIC0) and the percentage of time within mutant selection window (MSW) versus dosing interval (TMSW %) was subjected to Gaussian function and regression analysis. The results favored the consensus that time above MIC (T>MIC) was recognized as an important PK/PD parameter of cephalosporins for antibacterial efficiency. Cefquinome reached the maximum killing effect when T>MIC% attained approximately 40%~60%. The subsequent correlation analysis demonstrated that resistant S. aureus ATCC25923 was easy to occur when TMSW% attained an index of about 20% with t1/2β of 3 h after multiple dosing, and 40% with t1/2β of 6 h after multiple dosing

Spoilage Identification of Beef Using an Electronic Nose System

A commercially available Cyranose-320. conducting polymer-based electronic nose system was used to analyze the volatile organic compounds emanating from fresh beef strip loins (M. Longisimmus lumborum) stored at 4°C and 10°C. Two statistical techniques, i.e., linear discriminant analysis (LDA) and quadratic discriminant analysis (QDA), were used to develop classification models from the collected sensor signals. The performances of the developed models were validated by two different methods: leave-1-out cross-validation, and bootstrapping. The developed models classified meat samples based on the microbial population into “unspoiled” (microbial counts \u3c6.0 log10 cfu/g) and “spoiled” (microbial counts \u3e 6.0 log10 cfu/g). Overall, quadratic discriminant-based classification models performed better than linear discriminant analysis based models. For the meat samples stored at 10°C, the highest classification accuracies obtained by the LDA method with leave-1-out and bootstrapping validations were 87.10% and 85.87%, respectively. On the other hand, classification by QDA and subsequent validation by leave-1-out and bootstrapping provided highest accuracies of 87.5% and 97.38%, respectively. For samples stored at 4°C, the LDA method provided highest classification accuracies of 79.17% and 85.64% using leave-1-out and bootstrapping validation, respectively. When the QDA method was used, the highest classification accuracies obtained for the samples stored at 4°C were 87.50% and 98.48%, respectively, with leave-1-out and bootstrapping validations

Microdialysis Determination of Cefquinome Pharmacokinetics in Murine Thigh From Healthy, Neutropenic, and Actinobacillus pleuropneumoniae-Infected Mice

This study was aimed at applying microdialysis to explore cefquinome pharmacokinetics in thigh and plasma of healthy, neutropenic, and Actinobacillus pleuropneumoniae-infected mice. The relative recoveries (RRs) were tested in vitro by dialysis and retrodialysis and in vivo by retrodialysis. ICR mice were randomly divided into four groups: H-40 (healthy mice receiving cefquinome at 40 mg/kg), H-160, N-40 (neutropenic mice), and I-40 mg/kg (thigh infected-mice with A. pleuropneumoniae). After cefquinome administration, plasma was collected by retro-orbital puncture and thigh dialysate was collected by using a microdialysis probe with Ringer’s solution at a perfusion rate of 1.5 μL/min. Plasma and thigh dialysate samples were assessed by HPLC–MS/MS and analyzed by a non-compartment model. The mean in vivo recoveries in the thigh were 39.35, 38.59, and 37.29% for healthy, neutropenic, and infected mice, respectively. The mean plasma protein-binding level was 16.40% and was independent of drug concentrations. For all groups, the mean values of the free AUCinf in plasma were higher than those in murine thigh, while the elimination T1/2β for plasma were lower than those for murine thigh. Cefquinome penetration (AUCthigh/AUCplasma) from the plasma to thigh was 0.76, 0.88, 0.47, and 0.98 for H-40, N-40, I-40, and H-160 mg/kg, respectively. These results indicated that infection significantly affected cefquinome pharmacokinetics in murine thigh. In conclusion, we successfully applied a microdialysis method to evaluate the pharmacokinetics of cefquinome in murine thigh of healthy, neutropenic, and A. pleuropneumonia-infected mice and the pharmacokinetics of cefquinome was obviously affected by infection in thigh

Pharmacokinetic/Pharmacodynamic Integration to Evaluate the Changes in Susceptibility of Actinobacillus pleuropneumoniae After Repeated Administration of Danofloxacin

To evaluate the relationship between pharmacokinetic/pharmacodynamic (PK/PD) parameters and changes in susceptibility and resistance frequency of Actinobacillus pleuropneumoniae CVCC 259, a piglet tissue cage (TC) infection model was established. After A. pleuropneumoniae populations maintained at 108 CFU/mL in TCs, piglets were treated with various doses of danofloxacin once daily for 5 consecutive days by intramuscular injection. Both the concentrations of danofloxacin and the population of vial cells were determined. Changes in susceptibility and resistance frequency were monitored. Polymerase chain reaction (PCR) amplification of quinolone resistance-determining regions (QRDRs) and DNA sequencing were performed to identify point mutations in gyrA, gyrB, parC, and parE genes. Furthermore, the susceptibility of mutants to danofloxacin and enrofloxacin was determined in the presence or absence of reserpine to assess whether the mutants were caused by efflux pumps. The MICs and resistant frequency of A. pleuropneumoniae both increased when danofloxacin concentrations fluctuated between MIC99 (0.05 μg/mL) and MPC (mutant prevention concentration, 0.4 μg/mL). As for PK/PD parameters, the resistant mutants were selected and enriched when AUC24h/MIC99 ranged from 34.68 to 148.65 h or AUC24h/MPC ranged from 4.33 to 18.58 h. Substitutions of Ser-83→Tyr or Ser-83→Phe in gyrA and Lys-53→Glu in parC were observed. The susceptibility of mutants obtained via danofloxacin treatment at 1.25 and 2.5 mg/kg were less affected by reserpine. These results demonstrate that maintaining the value of AUC24h/MPC above 18.58 h may produce a desirable antibacterial effect and protect against A. pleuropneumoniae resistance to danofloxacin

Would you choose to be a psychiatrist again? A large-sample nationwide survey of psychiatrists and psychiatry residents in China

Abstract Background The mental health workforce sustainability in China suffers high rates of attrition and the intention to leave. Among current professionals, the intention to choose the same career is an interesting way to gauge their job satisfaction and other factors, and it may affect the career choices of younger generations. We aimed to survey the intention of psychiatrists and psychiatry residents to choose the same career if they could start over and to identify associated factors. Methods We conducted an anonymous survey of psychiatrists in 41 tertiary psychiatric hospitals in China. We collected demographic data, work-related information, the sense of professional identity, job satisfaction, and burnout (Maslach Burnout Inventory), and we specifically asked each participant whether they would choose to be a psychiatrist again if they could. Results Among 3,783 psychiatrists we surveyed, one-quarter responded that they would not choose to be a psychiatrist again if they had a choice, with less than half (47.2%) saying they would. Those who would not choose psychiatry again were more likely to have a negative (relative to positive) professional identity (OR = 7.47, P＜0.001, 95%CI: 4.587–12.164); experience job burnout (OR = 2.945, P＜0.001, 95%CI: 2.356–3.681); be dissatisfied with their job (OR = 2.739, P＜0.001, 95%CI: 2.102–3.569) and excessive regulation (OR = 1.819, P＜0.001, 95%CI: 1.487–2.226); have a heavy workload (OR = 1.749, P＜0.001, 95%CI: 1.423–2.149) or a lower income (OR = 1.748, P＜0.001, 95%CI: 1.415–2.161); be married (relative to single) (OR = 1.604, P = 0.004, 95%CI: 1.165–2.208); be dissatisfied with strained doctor-patient relationship (OR = 1.333, P = 0.005, 95%CI: 1.089–1.632); have more night shifts per month (OR = 1.055, P = 0.021, 95%CI: 1.008–1.104) or work longer hours per week (OR = 1.016, P = 0.001, 95%CI: 1.006–1.025). Conclusion Among psychiatrists in tertiary hospitals in China, those with a heavier workload, poor sense of professional identity, job dissatisfaction, and burnout were less likely to choose psychiatry again. Policymakers and hospital administrators need to take effective measures to improve psychiatrists’ sense of professional identity and increase their intention to stay

The PK/PD Integration and Resistance of Tilmicosin against Mycoplasma hyopneumoniae

Mycoplasma hyopneumoniae is the major pathogen causing enzootic pneumonia in pigs. M. hyopneumoniae infection can lead to considerable economic losses in the pig-breeding industry. Here, this study established a first-order absorption, one-compartment model to study the relationship between the pharmacokinetics/pharmacodynamics (PK/PD) index of tilmicosin against M. hyopneumoniae in vitro. We simulated different drug concentrations of timicosin in the fluid lining the lung epithelia of pigs. The minimum inhibitory concentration (MIC) of tilmicosin against M. hyopneumoniae with an inoculum of 106 CFU/mL was 1.6 μg/mL using the microdilution method. Static time–kill curves showed that if the drug concentration >1 MIC, the antibacterial effect showed different degrees of inhibition. At 32 MIC, the amount of bacteria decreased by 3.16 log10 CFU/mL, thereby achieving a mycoplasmacidal effect. The M. hyopneumoniae count was reduced from 3.61 to 5.11 log10 CFU/mL upon incubation for 96 h in a dynamic model with a dose of 40–200 mg, thereby achieving mycoplasmacidal activity. The area under the concentration-time curve over 96 h divided by the MIC (AUC0–96 h/MIC) was the best-fit PK/PD parameters for predicting the antibacterial activity of tilmicosin against M. hyopneumoniae (R2 = 0.99), suggesting that tilmicosin had concentration-dependent activity. The estimated value for AUC0–96 h/MIC for 2log10 (CFU/mL) reduction and 3log10 (CFU/mL) reduction from baseline was 70.55 h and 96.72 h. Four M. hyopneumoniae strains (M1–M4) with reduced sensitivity to tilmicosin were isolated from the four dose groups. The susceptibility of these strains to tylosin, erythromycin and lincomycin was also reduced significantly. For sequencing analyses of 23S rRNA, an acquired A2058G transition in region V was found only in resistant M. hyopneumoniae strains (M3, M4). In conclusion, in an in vitro model, the effect of tilmicosin against M. hyopneumoniae was concentration-dependent and had a therapeutic effect. These results will help to design the optimal dosing regimen for tilmicosin in M. hyopneumoniae infection, and minimize the emergence of resistant bacteria

Spoilage Identification of Beef Using an Electronic Nose System

A commercially available Cyranose-320. conducting polymer-based electronic nose system was used to analyze the volatile organic compounds emanating from fresh beef strip loins (M. Longisimmus lumborum) stored at 4°C and 10°C. Two statistical techniques, i.e., linear discriminant analysis (LDA) and quadratic discriminant analysis (QDA), were used to develop classification models from the collected sensor signals. The performances of the developed models were validated by two different methods: leave-1-out cross-validation, and bootstrapping. The developed models classified meat samples based on the microbial population into “unspoiled” (microbial counts 6.0 log10 cfu/g). Overall, quadratic discriminant-based classification models performed better than linear discriminant analysis based models. For the meat samples stored at 10°C, the highest classification accuracies obtained by the LDA method with leave-1-out and bootstrapping validations were 87.10% and 85.87%, respectively. On the other hand, classification by QDA and subsequent validation by leave-1-out and bootstrapping provided highest accuracies of 87.5% and 97.38%, respectively. For samples stored at 4°C, the LDA method provided highest classification accuracies of 79.17% and 85.64% using leave-1-out and bootstrapping validation, respectively. When the QDA method was used, the highest classification accuracies obtained for the samples stored at 4°C were 87.50% and 98.48%, respectively, with leave-1-out and bootstrapping validations.This article is from Transactions of the ASAE 47, no. 5 (2004): 1625–1633.</p

Determination of the Mutant Selection Window and Evaluation of the Killing of <i>Mycoplasma gallisepticum</i> by Danofloxacin, Doxycycline, Tilmicosin, Tylvalosin and Valnemulin

<div><p><i>Mycoplasma gallisepticum</i> is a common etiological cause of a chronic respiratory disease in chickens; its increasing antimicrobial resistance compromises the use of tetracyclines, macrolides and quinolones in the farm environment. Mutant selection window (MSW) determination was used to investigate the propensity for future resistance induction by danofloxacin, doxycycline, tilmicosin, tylvalosin and valnemulin. Killing of <i>M</i>. <i>gallisepticum</i> strain S6 by these antimicrobials was also studied by incubating <i>M</i>. <i>gallisepticum</i> into medium containing the compounds at the minimal concentration that inhibits colony formation by 99% (MIC₉₉) and the mutant prevention concentration (MPC). Based on the morphology and colony numbers of <i>M</i>. <i>gallisepticum</i> on agar plates, the four kinds of sera in the order of the applicability for culturing <i>M</i>. <i>gallisepticum</i> were swine serum > horse serum > bovine serum > mixed serum. The MPC/MIC₉₉ values for each agent were as follows: danofloxacin > tilmicosin > tylvalosin > doxycycline > valnemulin. MPC generated more rapid and greater magnitude killing than MIC₉₉ against <i>M</i>. <i>gallisepticum</i>. Under exposure of 10⁵–10⁹ CFU/mL at MPC drug levels, valnemulin had the slowest rate of reduction in viable organisms and danofloxacin had the highest rate of reduction.</p></div

Table_1_Nanopore-based metagenomic sequencing for the rapid and precise detection of pathogens among immunocompromised cancer patients with suspected infections.docx

Cancer patients are at high risk of infections and infection-related mortality; thereby, prompt diagnosis and precise anti-infectives treatment are critical. This study aimed to evaluate the performance of nanopore amplicon sequencing in identifying microbial agents among immunocompromised cancer patients with suspected infections. This prospective study enlisted 56 immunocompromised cancer patients with suspected infections. Their body fluid samples such as sputum and blood were collected, and potential microbial agents were detected in parallel by nanopore amplicon sequencing and the conventional culture method. Among the 56 body fluid samples, 47 (83.9%) samples were identified to have at least one pathogen by nanopore amplicon sequencing, but only 25 (44.6%) samples exhibited a positive finding by culture. Among 31 culture-negative samples, nanopore amplicon sequencing successfully detected pathogens in 22 samples (71.0%). Nanopore amplicon sequencing showed a higher sensitivity in pathogen detection than that of the conventional culture method (83.9% vs. 44.6%, P<0.001), and this advantage both existed in blood samples (38.5% vs. 0%, P=0.039) and non-blood samples (97.7% vs. 58.1%, P<0.001). Compared with the culture method, nanopore amplicon sequencing illustrated more samples with bacterial infections (P<0.001), infections from fastidious pathogens (P=0.006), and co-infections (P<0.001). The mean turnaround time for nanopore amplicon sequencing was about 17.5 hours, which was shorter than that of the conventional culture assay. This study suggested nanopore amplicon sequencing as a rapid and precise method for detecting pathogens among immunocompromised cancer patients with suspected infections. The novel and high-sensitive method will improve the outcomes of immunocompromised cancer patients by facilitating the prompt diagnosis of infections and precise anti-infectives treatment.</p