Reference Method Research Articles

Performance Evaluation of a Commercial Automated Library Preparation System for Clinical Microbial Whole-Genome Sequencing Assays

Next-generation sequencing (NGS) has proven clinical utility on disease management and serves as an important tool for genomic surveillance. Currently, hurdles surrounding its implementation, namely the complex and demanding analytical workflows, have impeded its widespread use in many laboratories. To address this challenge, the UCLA Molecular Microbiology and Pathogen Genomics Laboratory evaluated the performance of the Tecan MagicPrep NGS system, a commercial automated solution for library preparation for clinical whole-genome sequencing assays, against the Illumina Nextera DNA Flex Library Prep. Using 35 unique organisms (28 bacteria and 7 fungi) for various clinical applications, including microbial identification and genomic characterization, we compared the quantity and quality of the prepared libraries and the resulting sequences, and concordance of the overall results. We also assessed the impact of its implementation on laboratory workflow. The MagicPrep NGS produced higher library concentrations with smaller sizes, and correspondingly, higher molarity. Quality metrics of the sequences, however, demonstrated no significant impact on the overall results, producing 100% concordance with the reference method. Importantly, workflow analysis showed 5 hours less hands-on time per run with more flexibility. This evaluation study indicates that performance of the MagicPrep NGS is comparable to the Nextera DNA Flex with the added benefit of improving workflow efficiency and reducing labor for performing routine clinical microbial whole-genome sequencing tests.

New robust remote reference estimator using robust multivariate linear regression

SUMMARY The solution of the remote reference method, a frequently used technique in magnetotelluric (MT) data processing, can be viewed as a product of the two-input–multiple-output relationship between the local electromagnetic field and the reference field at a remote station. By applying a robust estimator to the two-input–multiple-output system, one can suppress the influence of outliers in the local magnetic field as well as those in the local electric field based on regression residuals. Therefore, this study develops a new robust remote reference estimator with the aid of robust multivariate linear regression. By applying the robust multivariate regression S-estimator to the multiple-output system, the present work derives a set of equations for robust estimates of the transfer function, noise variances, and the scale of the Mahalanobis distance simultaneously. The noise variances are necessary for the multivariate analysis to normalize the residuals of dependent variables. The Mahalanobis distance, a distance measure for multivariate data, is a commonly used indicator of outliers in multivariate statistics. By updating those robust estimates iteratively, the new robust remote reference estimator seeks the transfer function that minimizes the robust scale estimate of the Mahalanobis distance. The developed estimator can avoid bias in the MT transfer function even if there are significant noises in the reference magnetic field and handle outlying data more robustly than previously proposed robust remote reference estimators. The authors applied the developed method to a synthetic data set and real-world data. The test results demonstrate that the developed method downweights outliers in the local electric and magnetic fields and gives an unbiased transfer function.

A New Method to Minimize the Standard Deviation and Root Mean Square of the Prediction Error of Single-Optimized IOL Power Formulas.

The purpose of this study was to develop a simplified method to approximate constants minimizing the standard deviation (SD) and the root mean square (RMS) of the prediction error in single-optimized intraocular lens (IOL) power calculation formulas. The study introduces analytical formulas to determine the optimal constant value for minimizing SD and RMS in single-optimized IOL power calculation formulas. These formulas were tested against various datasets containing biometric measurements from cataractous populations and included 10,330 eyes and 4 different IOL models. The study evaluated the effectiveness of the proposed method by comparing the outcomes with those obtained using traditional reference methods. In optimizing IOL constants, minor differences between reference and estimated A-constants were found, with the maximum deviation at -0.086 (SD, SRK/T, and Vivinex) and -0.003 (RMS, PEARL DGS, and Vivinex). The largest discrepancy for third-generation formulas was -0.027mm (SD, Haigis, and Vivinex) and 0.002mm (RMS, Hoffer Q, and PCB00/SN60WF). Maximum RMS differences were -0.021 and +0.021, both involving Hoffer Q. Post-minimization, the largest mean prediction error was 0.726 diopters (D; SD) and 0.043 D (RMS), with the highest SD and RMS after adjustments at 0.529 D and 0.875 D, respectively, indicating effective minimization strategies. The study simplifies the process of minimizing SD and RMS in single-optimized IOL power predictions, offering a valuable tool for clinicians. However, it also underscores the complexity of achieving balanced optimization and suggests the need for further research in this area. The study presents a novel, clinically practical approach for optimizing IOL power calculations.

Analysis of Long-Distance Geometric Error Measurement and Uncertainty Based on PSD Laser Collimation Principle

Due to the limitations of traditional geometric error measurement, the measurement accuracy of long-stroke geometric errors is generally not high and the operation is complicated. In response to the above situation, in this study, a geometric error measurement system is built with a laser beam as the reference line and 2D position sensitive detector as the photoelectric conversion device. The single measurement range is 40 m, and the measurement range is further expanded through the principle of segmented splicing. Using an ultra-long guide rail as the measurement object for straightness measurement, the experimental results are similar to those of a laser interferometer. The uncertainty analysis model was obtained through the analysis of quantity characteristics, and based on this, the variance synthesis theorem and probability distribution propagation principle were studied to form two uncertainty synthesis methods. The measurement evaluation results showed that the two methods were basically consistent. The work provided a reference method for the uncertainty evaluation of position-sensitive detector measurement systems in the future.

Evaluation of left ventricular filling pressure by echocardiography in patients with atrial fibrillation

BackgroundEchocardiography is widely used to evaluate left ventricular (LV) diastolic function in patients suspected of heart failure. For patients in sinus rhythm, a combination of several echocardiographic parameters can differentiate between normal and elevated LV filling pressure with good accuracy. However, there is no established echocardiographic approach for the evaluation of LV filling pressure in patients with atrial fibrillation. The objective of the present study was to determine if a combination of several echocardiographic and clinical parameters may be used to evaluate LV filling pressure in patients with atrial fibrillation.ResultsIn a multicentre study of 148 atrial fibrillation patients, several echocardiographic parameters were tested against invasively measured LV filling pressure as the reference method. No single parameter had sufficiently strong association with LV filling pressure to be recommended for clinical use. Based on univariate regression analysis in the present study, and evidence from existing literature, we developed a two-step algorithm for differentiation between normal and elevated LV filling pressure, defining values ≥ 15 mmHg as elevated. The parameters in the first step included the ratio between mitral early flow velocity and septal mitral annular velocity (septal E/e’), mitral E velocity, deceleration time of E, and peak tricuspid regurgitation velocity. Patients who could not be classified in the first step were tested in a second step by applying supplementary parameters, which included left atrial reservoir strain, pulmonary venous systolic/diastolic velocity ratio, and body mass index. This two-step algorithm classified patients as having either normal or elevated LV filling pressure with 75% accuracy and with 85% feasibility. Accuracy in EF ≥ 50% and EF < 50% was similar (75% and 76%).ConclusionsIn patients with atrial fibrillation, no single echocardiographic parameter was sufficiently reliable to be used clinically to identify elevated LV filling pressure. An algorithm that combined several echocardiographic parameters and body mass index, however, was able to classify patients as having normal or elevated LV filling pressure with moderate accuracy and high feasibility.

Sedimentation field-flow fractionation for rapid phenotypic antimicrobial susceptibility testing: a pilot study.

The increase in antibiotic resistance is a major public health issue. The development of rapid antimicrobial susceptibility testing (AST) methods is becoming a priority to ensure early and appropriate antibiotic therapy. To evaluate sedimentation field-flow fractionation (SdFFF) as a method for performing AST in less than 3 h. SdFFF is based on the detection of early biophysical changes in bacteria, using a chromatographic-type technology. One hundred clinical Escherichia coli strains were studied. A calibrated bacterial suspension was incubated for 2 h at 37°C in the absence (untreated) or presence (treated) of five antibiotics used at EUCAST breakpoint concentrations. Bacterial suspensions were then injected into the SdFFF machine. For each E. coli isolate, retention times and elution profiles of antibiotic-treated bacteria were compared with retention times and elution profiles of untreated bacteria. Algorithms comparing retention times and elution profiles were used to determine if the strain was susceptible or resistant. Performance evaluation was done according to CLSI and the ISO standard 20776-2:2021 with broth microdilution used as the reference method. AST results from SdFFF were obtained in less than 3 h. SdFFF showed high categorical agreement (99.8%), sensitivity (99.5%) and specificity (100.0%) with broth microdilution. Results for each antimicrobial were also in agreement with the ISO 20776-2 recommendations, with sensitivity and specificity of ≥95.0%. This study showed that SdFFF can be used as a rapid, accurate and reliable phenotypic AST method with a turnaround time of less than 3 h.

Cross-Modality Reference and Feature Mutual-Projection for 3D Brain MRI Image Super-Resolution.

High-resolution (HR) magnetic resonance imaging (MRI) can reveal rich anatomical structures for clinical diagnoses. However, due to hardware and signal-to-noise ratio limitations, MRI images are often collected with low resolution (LR) which is not conducive to diagnosing and analyzing clinical diseases. Recently, deep learning super-resolution (SR) methods have demonstrated great potential in enhancing the resolution of MRI images; however, most of them did not take the cross-modality and internal priors of MR seriously, which hinders the SR performance. In this paper, we propose a cross-modality reference and feature mutual-projection (CRFM) method to enhance the spatial resolution of brain MRI images. Specifically, we feed the gradients of HR MRI images from referenced imaging modality into the SR network to transform true clear textures to LR feature maps. Meanwhile, we design a plug-in feature mutual-projection (FMP) method to capture the cross-scale dependency and cross-modality similarity details of MRI images. Finally, we fuse all feature maps with parallel attentions to produce and refine the HR features adaptively. Extensive experiments on MRI images in the image domain and k-space show that our CRFM method outperforms existing state-of-the-art MRI SR methods.

ANALYSIS OF SUPPLY CHAIN PERFORMANCE AND ADDED VALUE OF TEMPEH IN MATARAM CITY

Supply chain performance measurement presents a valuable opportunity to enhance and advance supply chain management practices across various industries, including the tempeh agroindustry. The research was aimed at analyzing the structure and performance of the supply chain, as well as the added value of the tempeh agroindustry in Mataram city. The study utilized a descriptive method that involved interviewing several respondents. Collected data was analyzed using the Supply Chain Operation Reference (SCOR) method version 9, and Hayami Model Added Value Analysis. The results indicated that the stakeholders involved in the agroindustry supply chain in Mataram city were soybean importers in Java, local soybean supply agents, tempeh agroindustry entrepreneurs, and tempeh retailers. The study found that the tempeh agroindustry supply chain performance attained superior performance in the Reliability, Flexibility, Responsiveness, and Asset attributes. On the other hand, the Total Supply Chain Cost attribute recorded poor performance. Processing one kilogram of soybeans into tempeh yielded an added value of IDR. 9,280.82, with an added value ratio of 43.56%, and is categorized as moderate.

Bacteriuria and phenotypic antimicrobial susceptibility testing in 45 min by point-of-care Sysmex PA-100 System: first clinical evaluation

PurposeThis study compared the results of the new Sysmex PA-100 AST System, a point-of-care analyser, with routine microbiology for the detection of urinary tract infections (UTI) and performance of antimicrobial susceptibility tests (AST) directly from urine.MethodsNative urine samples from 278 female patients with suspected uncomplicated UTI were tested in the Sysmex PA-100 and with reference methods of routine microbiology: urine culture for bacteriuria and disc diffusion for AST.ResultsThe analyser delivered bacteriuria results in 15 min and AST results within 45 min. Sensitivity and specificity for detection of microbiologically confirmed bacteriuria were 84.0% (89/106; 95% CI: 75.6–90.4%) and 99.4% (155/156; 95% CI: 96.5–100%), respectively, for bacterial species within the analyser specifications. These are Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterococcus faecalis and Staphylococcus saprophyticus, which are common species causing uncomplicated UTI. Overall categorical agreement (OCA) for AST results for the five antimicrobials tested in the Sysmex PA-100 (amoxicillin/clavulanic acid, ciprofloxacin, fosfomycin, nitrofurantoin and trimethoprim) ranged from 85.4% (70/82; 95%CI: 75.9–92.2%) for ciprofloxacin to 96.4% (81/84; 95% CI: 89.9–99.3%) for trimethoprim. The Sysmex PA-100 provided an optimal treatment recommendation in 218/278 cases (78.4%), against 162/278 (58.3%) of clinical decisions.ConclusionThis first clinical evaluation of the Sysmex PA-100 in a near-patient setting demonstrated that the analyser delivers phenotypic AST results within 45 min, which could enable rapid initiation of the correct targeted treatment with no further adjustment needed. The Sysmex PA-100 has the potential to significantly reduce ineffective or unnecessary antibiotic prescription in patients with UTI symptoms.

Diagnostic Strategy Using Color Doppler Ultrasound of Temporal Arteries in Patients With High Clinical Suspicion of Giant Cell Arteritis : A Prospective Cohort Study.

Giant cell arteritis (GCA) is the most prevalent systemic vasculitis in people older than 50 years. Any delay in diagnosis impairs patients' quality of life and can lead to permanent damage, particularly vision loss. To evaluate a diagnostic strategy for GCA using color Doppler ultrasound of the temporal artery as a first-line diagnostic test, temporal artery biopsy (TAB) as a secondary test, and physician expertise as the reference method. Prospective multicenter study with a 2-year follow-up. (ClinicalTrials.gov: NCT02703922). Patients were referred by their general practitioner or ophthalmologist to a physician with extensive experience in GCA diagnosis and management in one of the participating centers: 4 general and 2 university hospitals. 165 patients with high clinical suspicion of GCA, aged 79 years (IQR, 73 to 85 years). The diagnostic procedure was ultrasound, performed less than 7 days after initiation of corticosteroid therapy. Only ultrasound-negative patients underwent TAB. Bilateral temporal halo signs seen on ultrasound were considered positive. Ultrasound and TAB results were compared with physician-diagnosed GCA based on clinical findings and other imaging. Diagnosis of GCA was confirmed in 44%, 17%, and 21% of patients by ultrasound, TAB, and clinical expertise and/or other imaging tests, respectively. Their diagnosis remained unchanged at 1 month, and 2 years for those with available follow-up data. An alternative diagnosis was made in 18% of patients. The proportion of ultrasound-positive patients among patients with a clinical GCA diagnosis was 54% (95% CI, 45% to 62%). Small sample size, no blinding of ultrasound and TAB results, lack of an objective gold-standard comparator, and single diagnostic strategy. By using ultrasound of the temporal arteries as a first-line diagnostic tool in patients with high clinical suspicion of GCA, further diagnostic tests for patients with positive ultrasound were avoided. Tender "Recherche CH-CHU Poitou-Charentes 2014."

Performance analysis of supply chain for construction materials at high rise building project based on Supply Chain Operations Reference Method: A case study

Abstract Supply chain management is an approach to implementing lean construction efforts to minimize construction project problems, such as cost overruns, project delays, and disputes. Implementation of supply chain management is necessary, especially in the procurement of materials. However, there is a need to measure performance during the implementation. Results of measurement can show the extent to which the project operates and use it as a benchmark to determine the end impact of a project. This study’s purpose is to measure of supply chain performance with the supply chain operation reference (SCOR) method and use the analytical hierarchy process (AHP). Compile data using an observation with scoring form based on standards adoption, categories, and SCOR metrics through key performance indicators (KPI) and structured interviews with the contractor to validate. Limited to the study is the supply chain of ready-mix concrete materials. The AHP method give the weight of the output value of the KPI. The results of the KPI measurements from the contractor’s perspective are that one indicator is still at a low value, which is a cycle time for fulfilling orders. An overall result of 84.83 from the measurement KPI suggests a “good” performance category.

Continuous wavelet transform and integration of discrete wavelet transform with principal component analysis and fuzzy inference system for the simultaneous determination of ethinyl estradiol and drospirenone in combined oral contraceptives

In this study, the spectrophotometric method integrated with continuous wavelet transform (CWT) and coupled discrete wavelet transform (DWT) with fuzzy inference system (FIS) was developed for the simultaneous determination of ethinyl estradiol (EE) and drospirenone (DP) in combined oral contraceptives (COCs). The CWT approach was performed in the linearity range of 0.6–6 µg/mL for EE and 0.9 to 18 µg/mL for DP. Biorthogonal with an order of 1.3 (bior1.3) at a wavelength of 216 nm and Daubechies with an order of 2 (db2) at a wavelength of 278 nm were selected as the best wavelet families for obtaining the best zero crossing point for EE and DP, respectively. The limit of detection (LOD) of 0.7677 and 0.3222 µg/mL and the limit of quantification (LOQ) of 2.326 and 0.9765 µg/mL were obtained for EE and DP, respectively. The mean recovery of 103.24% and 99.77%, as well as root mean square error (RMSE) of 0.1896 and 0.1969, were found for EE and DP, respectively. In the DWT, the absorption of the mixtures was decomposed using different wavelets named db4, db2, Symlet2 (sym2), and bior1.3. Each of the wavelet outputs was dimension reduced by the principal component analysis (PCA) method and considered as FIS input. The wavelet of db4 with the coefficient of determination (R2) of 0.9979, RMSE of 0.0968, and mean recovery of 100.63% was chosen as the best one for the EE, while bior1.3 with R2 of 0.9955, RMSE of 0.4055, and mean recovery of 101.93% was selected for DP. These methods were successfully used to analyze the EE and DP simultaneously in tablet pharmaceutical formulation without any separation step. The suggested methods were compared with a reference method (HPLC) using analysis of variance (ANOVA) at a 95% confidence level, and no significant difference was observed in terms of accuracy. The suggested chemometric methods are reliable, rapid, and inexpensive, and can be used as an environmentally friendly alternative to HPLC for the simultaneous estimation of the mentioned drugs in commercial pharmaceutical products.

Establishment and evaluation of a new fluorescent probe method based on loop-mediated isothermal amplification for the detection of Mycobacterium tuberculosis complex.

We aimed to develop a novel diagnostic method called multiplex fluorescence of loop primer upon self-dequenching loop-mediated isothermal amplification (mFLOS-LAMP) for the rapid detection of Mycobacterium tuberculosis complex (MTBC). A set of specific primers was designed to target the detection of IS1081 and IS6110 genes, which are insertion sequences within the MTBC. The 110 sputum specimens collected were assessed using the established mFLOS-LAMP method, multiplex polymerase chain reaction, Xpert MTB/RIF, and smear microscopy. The optimal reaction temperature and duration for mFLOS-LAMP were determined to be 65°C and 30 min, respectively, by optimizing the entire system. The detection sensitivity of mFLOS-LAMP was 6.0 × 101 CFU/mL, by Bacillus Calmette-Guerin, and the mFLOS-LAMP sensitivity of M. tuberculosis H37Rv genomic DNA was 500 fg, and the specificity was 100%. The sensitivity of mFLOS-LAMP was 94.2% and the specificity was 96.6%, when Xpert MTB/RIF was used as the reference method. There was no statistically significant difference in their detection rate (χ2 = 0, P = 1.000), and the consistency was good (kappa = 0.909, P < 0.001). The receiver operating characteristic analysis yielded the maximum area under the curve of 0.954. The mFLOS-LAMP method demonstrated high sensitivity and specificity, allowing for swift and accurate detection of MTBC.

Meteorological measurements affected by wind farm cluster wakes: How to deal with waked wind in AEP assessments?

This research explores a spectrum of correction methods to predict free wind speed conditions from offshore wind measurements influenced by nearby wind farm clusters. Three of these methods (reference data, wake model and mesoscale model correction) were evaluated at two measurement sites in the German Bight, and a matrix of wake correction factors was obtained for each method. Comparison among methods reveals that all can identify sectors affected by upstream clusters. However, using reference data for correction overestimates impacts at sector limits. Wake and mesoscale model corrections are similar in their proposed correction factors, but the wake method yields lower values due to underestimation of wind farm wake deficits over long distances. When applying the correction factors to the waked measurements, the resulting series did not show significant statistical differences. The choice of correction method is contingent upon factors such as time constraints, availability of information on surrounding wind farms, and desired level of accuracy. The reference method is suggested in the absence of relevant information, while the wake method is recommended when there are time constraints. The mesoscale method can be considered when more time is available.

Chemiluminescence data modeling using parallel factor analysis for the simple and sensitive determination of diphenoxylate in human plasma and tablets

AbstractIn this study, a chemiluminescence (CL) method was developed to determine diphenoxylate in tablets and human plasma. This is the first CL method proposed to determine diphenoxylate. Creating three‐dimensional data caused the parallel factor analysis algorithm (PARAFAC) to be used for the first time in CL methods. The method is based on the fact that diphenoxylate enhances the weak CL produced in the reaction of Ru(phen)32+ and acidic Ce(IV), and the concentration of Ce(IV) solution has a different effect on the CL response of diphenoxylate and the blank plasma. The calibration curve was linear from 4.0 × 10−8 to 1.6 × 10−6 mol L−1 (R2 = 0.9954), and the detection limit was 1.3 × 10−8 mol L−1 (S/N = 3). The sampling rate was about 30 samples per hour, and the % RSD for 10 repeated measurements of 4 × 10−7 mol L−1 diphenoxylate was 5.4%. The interference effects of some ions, amino acids, and common additives were also investigated. The CL method was successfully used to determine diphenoxylate in tablets, and the results were statistically confirmed by the reference method. The proposed CL method and the PARAFAC algorithm were successfully used to determine the concentration of diphenoxylate in human blood plasma samples.

Comparison of Colistin Susceptibility via Two Different Methods in Gram-Negative Extensive Drug-Resistance Isolates from ICU Patients.

To compare the susceptibility of colistin by two methods in extensive drug-resistant (XDR) Gram-negative isolates from ICU patients. Cross-sectional comparative analysis. Place and Duration of the Study: Department of Microbiology, Combined Military Hospital Karachi, Pakistan, from August 2022 to February2023. A total of 100 clinical specimens received from the intensive care unit yielded growth of extensively drug-resistant gram-negative bacteria, which were evaluated for polymyxin E susceptibility. The agar dilution method was compared with the reference broth microdilution (BMD) method. Minimum inhibitory concentration (MIC) was noted for both methods. Comparison of the MIC method by agar dilution showed a 90% correlation with the reference method of broth microdilution. With MICs within the acceptable range of the clinical and laboratory standards institute (CLSI) recommendations, 89 isolates were susceptible to colistin, whereas only 11 remained resistant. Polymyxin E's MIC 50 and MIC 90 were determined to be 1 and 2 µg/ml, respectively, with 97% susceptibility. Agar dilution susceptibility method can be used for screening purposes for the susceptibility testing of polymyxin E. This method is reliable and can easily identify the heteroresistance. Extensively drug-resistant, Broth microdilution, Multidrug-resistant, Agar dilution, Minimum inhibitory concentration, Colony forming unit.

Rapid detection of Ebolavirus using isothermal recombinase-aided amplification.

Ebolavirus disease (EVD) is an often-lethal disease caused by the genus Ebolavirus (EBOV). Although vaccines are being developed and recently used, outbreak control still relies on a combination of various factors, including rapid identification of EVD cases. This allows rapid patient isolation and control measure implementation. Ebolavirus diagnosis is performed in treatment centers or reference laboratories, which usually takes a few hours to days to confirm the outbreak or deliver a clear result. A fast and field-deployable molecular detection method, such as the isothermal amplification recombinase-aided amplification (RAA), could significantly reduce sample-to-result time. In this study, a RT-RAA assay was evaluated for EBOV detection. Various primer and probe combinations were screened; analytical sensitivity and cross-specificity were tested. A total of 40 archived samples from the 2014 to 2016 Ebola outbreak in West Africa were tested with both the reference method real-time RT-PCR and the established RT-RAA assay. The assay could detect down to 22.6 molecular copies per microliter. No other pathogens were detected with the Ebolavirus RT-RAA assay. Testing 40 samples yield clinical sensitivity and specificity of 100% each. This rapid isothermal RT-RAA assay can replace the previous RT-RPA and continue to offer rapid EBOV diagnostics.

Predicting the chemical composition of pet food with hyperspectral imaging

Performing fast analyses to control the nutritional quality of pet food products is of great interest in the pet food industry. The purpose of this study was to test the efficacy of near-infrared hyperspectral imaging (HSI) in determining chemical composition content of cat and dog food, and evaluate the influence of mixed model and characteristic wavelength on the performance of the quantitative model. Seventy cat food and thirty-six dog food samples were characterised using reference methods for moisture, crude protein (CP), crude fat (CF), crude fibre (CFe), crude ash (CA), calcium (Ca) and total phosphorus (TP) content. The Partial Least Squares regression (PLSR) was used to establish the quantitative models that involved the cat food model and mixed model. The characteristic wavelength was selected using a competitive adaptive reweighted-sampling (CARS) algorithm. Results showed that, except for CFe, the performance of the cat food model was similar to the mixed model. When using feature band, the prediction results of the mixed models for CP, CF, moisture, and CFe were optimized with R2p between 0.73 and 0.96 and RPD in the range 2.22–5.20, but the prediction results of TP, CA, and Ca did not meet the actual demand. Optimal quantitative models were used to visualise the sample chemical composition distributions. The results of this study provide theoretical and technical support for the rapid online quality control of pet food.

Evaluation of Automated Disk Diffusion Antimicrobial Susceptibility Testing Using Radian® In-Line Carousel

Antimicrobial susceptibility testing (AST) by disk diffusion provides an accurate image of bacterial growth, enabling the detection of culture purity, heterogeneous growth, and antibiotic interactions. However, this manual method is time-consuming and visual interpretation is prone to errors. To overcome these disadvantages, the Radian® In-Line Carousel (Copan, Brescia, Italy) was launched, which is a WASPLab® module dedicated to full automation of (pre)-analytical steps as well as interpretation of disk diffusion AST. However, until now, no evaluation of Radian® against manual disk diffusion has been performed. We assessed the categorical agreement (CA) between standardized disk diffusion (reference method) and Radian® using EUCAST 2021 breakpoints. We tested 135 non-duplicate strains, selected from the National EUCAST challenge panel, clinical strains, and external quality controls. The strains included Enterobacterales (n = 63), Enterococcus faecalis (n = 3), Enterococcus faecium (n = 10), Pseudomonas aeruginosa (n = 16), Staphylococcus aureus (n = 19), coagulase-negative staphylococci (n = 4), and Streptococcus spp. (n = 20). Furthermore, we explored antibiotic disk thermolability in the WASP Radian® carousel by testing 10 ATCC® strains up to 7 days. The observed CA was 95.3%, 96.3%, 93.8%, 97.3% and 98.0% for Enterobacterales, Enterococcus spp., P. aeruginosa, Staphylococcus spp. and Streptococcus spp., respectively, resulting in an acceptable overall CA for all groups. (Very) major error rates were ≤ 5% for all antibiotics. Antibiotic disk thermostability was confirmed up to 4 days in the WASP Radian® In-Line Carousel. The Radian® In-Line Carousel provides a fully automated solution for accurate disk diffusion AST, reducing workload and improving standardization and traceability. In addition, our study demonstrated the thermostability of antibiotic disks up to 4 days in the WASP Radian® In-Line Carousel.

Real-Time Monitoring of Odour Emissions at the Fenceline of a Waste Treatment Plant by Instrumental Odour Monitoring Systems: Focus on Training Methods.

Waste treatment plants (WTPs) often generate odours that may cause nuisance to citizens living nearby. In general, people are becoming more sensitive to environmental issues, and particularly to odour pollution. Instrumental Odour Monitoring Systems (IOMSs) represent an emerging tool for continuous odour measurement and real-time identification of odour peaks, which can provide useful information about the process operation and indicate the occurrence of anomalous conditions likely to cause odour events in the surrounding territories. This paper describes the implementation of two IOMSs at the fenceline of a WTP, focusing on the definition of a specific experimental protocol and data processing procedure for dealing with the interferences of humidity and temperature affecting sensors' responses. Different approaches for data processing were compared and the optimal one was selected based on field performance testing. The humidity compensation model developed proved to be effective, bringing the IOMS classification accuracy above 95%. Also, the adoption of a class-specific regression model compared to a global regression model resulted in an odour quantification capability comparable with those of the reference method (i.e., dynamic olfactometry). Lastly, the validated models were used to process the monitoring data over a period of about one year.