Analysis and Appraisal of Fascine in Shahe Ancient Bridge Ruins, Xi’an, Shaanxi, China

The Chinese civilization has a long history, and the Chinese ancestors invented the "aquatic engineering technology" for flood control and water management as early as two thousand years ago: Fascine Body. The Fascine Body is a structure used to protect the bank and block the breach in the ancient Chinese flood control project. The excavation of the Shahe Ancient Bridge Ruins in Xi’an City, Shaanxi Province, China discovered the existence of a fascine body structure. Through C14 dating, fiber slice observation, infrared spectroscopy, X-ray diffraction, thermogravimetric analysis and SEM energy spectrum analysis of the fascine material, at the same time, conduct the microbial identification on it and the surrounding soil, analyze its dominant bacterial community, and control its microbial diseases in a targeted manner. The research on the fascine bank ruins solved the boundary problem of the width and length of the Shahe ancient bridge, evaluated the exact age of Shahe ancient bridge, provided the important materials for the research on ancient bridges, river embankments and other ruins, and also provided the important clues for the traffic and layout around Chang'an during the Qin and Han Dynasties of Chinese history.

SPECIFIC MICROBIOLOGY SPECTRUM OF WHITE SAND MUSSELS FROM KEY SAMPLE SPOTS FROM BULGARIAN BLACK SEA AQUATORY

The "white sand mussels" are edible bivalves inhabiting the littoral shores usually buried in the sand. Тhey are invasive species for the Bulgarian waters of the Black Sea. The samples for this study were collected from different points on the northern and southern Black Sea Bulgarian coast in the period January 2020 to December 2020. The study of different types of microorganisms was performed by using the microbial identification system model: MicroLog M® BIO45101 BiologInc and the software product GEN III. The physic-chemical parameters of the waters – temperature, pH, salinity and dissolved O2 were also determined. In the different species, we had detected specific microbiological complexes. The species Pseudomonas viridilivida and Citrobacter farmer were isolated only from Donax trunculus. The species Escherichia hermannii was found only in Mya arenaria, and Acinetobacter johnsonii was detected only in Chamelea gallina. The isolated species Acinetobacter gyllenbergii and Acinetobacter johnsonii are related to humans and are indicators for pollution of the water with channel waste waters. Our results demonstrated an increase ofhe quantity of the coliforms the region of Sveti Vlas from August, where they were 50 the norms. In the region of Arkutino in July and Ahtopol in August, the quantity of the fecal coliforms is 190 and 30 times the norms prescribed in the Ordinanceo. 4 from 20.10.2000 for the quality of fisheries water and the breeding of shellfish (the amount of fecal coliforms in the inter-shell content should be less than 300 NVB). We noticed also seriousollution of the Varna lake even months after an accident with a leaky pipe.

665. Clinical and Financial Impact of Next Generation Sequencing (NGS) in addition to Conventional Microbiology Testing in our Urban Referral Health Center

Background Clinical microbiology traditionally relies on culture methodology and serological testing, that have inherent limitations. Newer diagnostic techniques such as Next Generation Sequencing (NGS) have shown promise to improve microbial identification. In select scenarios, we send clinical specimens to reference laboratories for NGS testing in addition to current standard of care (SOC) diagnostics. We wanted to determine how this diagnostic approach has impacted patient care. We also wanted to review the financial burden through cost-benefit analysis for these ‘send-out’ tests. Methods We performed a retrospective chart review of all cases over a 3-year period in which NGS was submitted. Data, including demographics, comorbidities, antimicrobial use, and diagnosis (by SOC and NGS) were gathered. We delineated how often there was concordance or discordance between SOC and NGS. We also obtained information on financial cost (direct and indirect) and turnaround time (TAT) for NGS results. Results A total of 33 clinical specimens from 25 patients were sent for NGS. The majority of specimens comprised joint tissue/fluid, organ tissue and CSF. Concordance occurred between SOC and NGS testing in 75.8% (25/33) of samples; of those, 88% excluded infection. NGS identified a pathogen in 20% (5/25) patients in which concomitant SOC testing was negative. A subsequent change in antimicrobial management occurred in 16% (4/25) of patients. The mean TAT was 14 days and average cost per specimen was &821.52 (range: &573-&1590). Table 1. Pathogens identified by NGS with negative traditional microbiological test results Figure 1. Distribution of specimen site (in %) sent for NGS Conclusion NGS can provide additional diagnostic sensitivity in infectious diseases, which at our institution identified a new pathogen in 20% and a resultant treatment change in 16% of our patients. This testing may also allow physicians to reaffirm the absence of an infection diagnosis. A larger NGS testing population may reveal more significant benefits. While the attributable cost of NGS was substantial, it should be measured against the costs of administration of unnecessary antibiotics, inaccurate diagnosis, and adverse patient outcomes that may result from SOC testing alone. Considering its financial cost and extended TAT, in-house NGS testing may be warranted to facilitate a higher volume of testing. Disclosures All Authors: No reported disclosures

659. Correlate Clinically and More-Use of Interpretative Comments in Clinical Microbiology Reporting

Background Microbial identification & antibiotic susceptibility testing is an important investigation in clinical microbiology laboratory. In many centres in India the report has only the isolate and antibiotics tested. The additional comments if added give guidance to the clinicians to utilize the results. Pre-analytical issues of adequate & relevant clinical history, appropriate sampling techniques, timely transport & storage, history of antibiotic usage along with post analytical issues of recommended line of antibiotic therapy and infection control practices are better addressed with this practice. Methods This was a prospective qualitative study from the period of January 2017-March 2021 where in the standard operating protocol of Clinical Microbiology was reviewed and appropriate comments were included in the Laboratory Information System once the isolate was identified using VITEK 2, automated ID/AST instrument and interfaced. The Clinical Microbiologist would then review the comments upon discussion with the clinicians and then authorize reports. The reports included sample & isolate specific details , recommended antibiotic therapy and infection control related comments. This was based on standard international and national guidelines (CLSI, EUCAST, IDSA, IAP, and National Treatment Guidelines of India). Results There was a gradual improvement in completion of request forms with clinical history, sample site and antibiotic history being mentioned. This was assessed through periodic audits conducted every quarter from 36% in March 2017 to 95% in March 2021. Clinical communication with the microbiology laboratory also showed improvement with documentation. Feedback from clinicians was also taken on the utility of these comments, (87/120)72.5% of the clinicians found them useful(Grade 5). (32/120) 26 %(Grade 3) of the clinicians had concerns about the turnaround time and requested for provisional reports. Conclusion Interpretative comments in reports act as a bridge between clinical microbiology, infectious diseases and infection control. They help us to choose the correct antibiotics or sometimes no antibiotics when the situation demands it. With all the recent advancements, the clinico-microbiological utility of culture reports is the need of the hour. Disclosures All Authors: No reported disclosures

Ribosomal protein database profiling lends clarity to ribosomal protein evolution and mass distribution

Existence of theoretical ribosomal protein mass fingerprint as well as utility of ribosomal protein as biomarkers in mass spectrometry microbial identification suggests phylogenetic significance for this class of proteins. To serve the above two functions, facile means of identifying and extracting important attributes of ribosomal proteins from proteome data file of microbial species must be found. Additionally, there is a need to calculate important properties of ribosomal proteins such as molecular weight and nucleotide sequence based on amino acid sequence information from FASTA proteome file. This work sought to support the above endeavour through developing a MATLAB software that extracts the amino acid sequence information of all ribosomal proteins from the FASTA proteome datafile of a microbial species downloaded from UniProt. Built-in functions in MATLAB are subsequently employed to calculate important properties of extracted ribosomal proteins such as number of amino acid residue, molecular weight and nucleotide sequence. All information above are output, as a database, to an Excel file for ease of storage and retrieval. Data available from the analysis of an Escherichia coli K-12 proteome revealed that the bacterium possess a total of 59 ribosomal proteins distributed between the large and small ribosome subunits. The ribosomal protein ranges in sequence length from 38 (50S ribosomal protein L36) to 557 (30S ribosomal protein S1). In terms of molecular weight distribution, the profiled ribosomal proteins range in weight from 4364.305 Da (50S ribosomal protein L36) to 61157.66 Da (30S ribosomal protein S1). More important, analysis of the distribution of the molecular weight of different ribosomal proteins in E. coli reveals a smooth curve that suggests strong co-evolution of ribosomal protein sequence and mass given the tight constraints that a functional ribosome presents. Finally, cluster analysis reveals a preponderance of small ribosomal proteins compared to larger ones, which remains to be a mystery to evolutionary biologists. Overall, the information encapsulated in the ribosomal protein database should find use in gaining a better appreciation for the molecular weight distribution of ribosomal proteins in a species, as well as delivering information for using ribosomal protein biomarkers in identifying particular microbial species in mass spectrometry microbial identification.

Tejuino, a Traditional Fermented Beverage: Composition, Safety Quality, and Microbial Identification

This study aims to analyze the chemical and microbial composition and characterize volatile compounds from the artisanal and commercial Tejuino beverage. For this, eight samples are analyzed (four artisanal and four commercial). The chemical and microbiological quality is determined by standard methods, and volatile compounds are determined by solid-phase microextraction. Overall, the physicochemical composition and microbiological quality are higher for artisanal Tejuino (p < 0.05). The pH values were 3.20 and 3.62, and 0.76 and 0.46 meq of lactic acid for artisanal and commercial Tejuino, respectively. With volatile compounds analyzed, esters, benzenes, and aldehydes were predominant; meanwhile, ethanol was a volatile compound with the highest concentration for all samples. Saccharomyces cerevisiae and Limosilactobacillus fermentum were identified in artisanal Tejuino; yeasts of the Pichia genera and Lactiplantibacillus plantarum, for commercial Tejuino, and Enterococcus genus were identified in both samples. The characterization of both types of Tejuino allows us to update the information available on this important Mexican beverage. In addition, the isolation of lactic acid bacteria, as representative bacteria of both drinks, offers an area of opportunity to know the potential functionality of these bacteria in traditional fermented products.

Development and Validation of a MALDI-TOF-Based Model to Predict Extended-Spectrum Beta-Lactamase and/or Carbapenemase-Producing in Klebsiella pneumoniae Clinical Isolates

Objectives: MALDI-TOF Mass Spectrometry (MS) is a reference method for microbial identification at clinical microbiology laboratories. We have designed and validated a new multiview model based on machine learning from MS spectra to predict antibiotic resistance mechanisms 24 h before phenotypic results are available. Methods: Antibiotic susceptibility of 402 clinical Klebsiella pneumoniae isolates was determined in two collections, discriminating among Wild Type (WT), Extended-Spectrum Beta-Lactamases (ESBL) producers, and ESBL and Carbapenemases (ESBL+CP) producers. Each isolate was subcultured 3 consecutive days and 2 independent spectra were acquired in each replica (6 MS spectra/isolate). Spectra were automatically classified by a kernelized Bayesian factor analysis model (KSSHIBA), using two independent strategies: 1) the model was designed with isolates from a single center and validated with isolates from the other center; and 2) in a second stage all isolates were used at the same time for design and validation processes. Results: Higher prediction values were obtained when integrating all isolates with hospital collection of origin information. Our model exhibited higher prediction capability than current state-of-the-art models, particularly in intercollection scenarios because local epidemiology could introduce relevant variables affecting prediction accuracy. Conclusions: Compared to previously reported studies, our model demonstrated the highest ability to predict ESBL and/or CP production in clinical K. pneumoniae isolates and it provided an efficient way to combine information from different centers. Its implementation in microbiological laboratories could improve the detection of multi-drug resistant isolates, optimizing the therapeutic decision.

Cost-effectiveness of rapid laboratory-based mass-spectrometry diagnosis of bloodstream infection: evidence from the RAPIDO randomised controlled trial

Objectives and interventionBloodstream infection, the presence of viable micro-organisms in the blood, is a prevalent clinical event associated with substantial mortality. Patient outcomes may be improved when the causative micro-organism is identified quickly. We assessed the cost-effectiveness of rapid microbial identification by matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry.DesignEconomic evaluation alongside a randomised multicentre trial (RAPIDO: RAPId Diagnosis on Outcome) assessing the impact of rapid identification by MALDI-TOF spectrometry.SettingAdult inpatients with bloodstream infections at seven National Health Service hospital trusts in England and Wales.Primary outcomeNet monetary benefit, estimated as incremental costs compared with incremental 28-day survival, of rapid identification by MALDI-TOF spectrometry compared with conventional identification.MethodsPatients were randomised (1:1) to receive diagnosis by conventional methods of microbial identification (conventional arm) only or by MALDI-TOF spectrometry in addition to conventional identification (RAPIDO arm).ResultsData from 5550 patients were included in primary analysis. Mean imputed costs in 2018/2019 prices per patient were lower by £126 in the RAPIDO arm (95% CI −£784 to £532) but the proportion of patients alive at day 28 was lower (81.4% vs 82.3%). The probability of cost-effectiveness of MALDI-TOF was <0.5 at cost-effectiveness thresholds between £20 000 and £50 000.ConclusionsAdjunctive MALDI-TOF diagnosis was unlikely to be cost-effective when measured as cost per death avoided at 28 days. However, the differences between arms in cost and effect were modest, associated with uncertainty and may not accurately reflect ‘real-world’ routine use of MALDI-TOF technology in this patient group.Trial registration numbersISRCTN97107018/UKCRN 11978.

Use of Blood Culture Bottles to Incubate Irrigation Water from the Surgical Site May Improve Detection of Causative Organisms in Pyogenic Vertebral Osteomyelitis

【Background】 Identification of pathogenic microorganisms are essential for pyogenic vertebral osteomyelitis (PVO). This study aimed to demonstrate the effectiveness of the blood culture bottle (BCB) system in identifying PVO causative organisms. 【Materials & Methods】 We analyzed retrospectively collected data from patients who underwent full-endoscopic spine surgery for PVO between January 2016 and March 2019. Irrigation water generated in the surgical field was incubated in the BCB system, and compared with blood culture before surgery and tissue culture taken by the conventional method. The microbial identification rate and the time from sample collection to microbial identification of the BCB system were compared with conventional blood culture and tissue cultures using the Wilcoxon signed-rank test. 【Results】 We included 17 patients (12 men, five women; mean age, 72.8 ± 11.9 years). Bacteria were cultured from 3 (17.6%), 13 (76.5%), and 12 (70.6%) patients by blood culture, tissue culture, and the BCB system, respectively. Tissue culture and the BCB system had significantly higher detection rates than blood culture (P = .002, P =.003), and there was no significant difference between tissue culture and BCB system (P = .655). In 15 cases (88.2%), the causative organism was identified by at least one method. In two cases, the causative organism (Escherichia coli) was only identified by the BCB system. The BCB system required amount of time for microbial identification (3.9 ± 3.0 days), compared with the time required for blood culture (5.0 ± 1.4 days, P =.180) and tissue culture (11.9 ± 15.1 days, P =.012). 【Conclusions】 Results suggest the possibility of improving the detection rate and time to detection of causative organisms by the BCB system as an adjunct to the conventional method.