Anaerobic Bottles Research Articles

Comparison of microbial growth on primed extracorporeal membrane oxygenation circuits in varying environments using different priming solutions.

Extracorporeal Membrane Oxygenation (ECMO) is a life support device for patients with severe heart and/or lung failure. Emergency situations require immediate ECMO response. Primed circuits have become a routine practice, as it may take 30-60min to assemble and prime. There remains a lack of data to support the sterility of primed and stored ECMO circuits. This bench study assessed the impact of storage environment and priming solution on specific microbial growth of primed ECMO circuits. Twelve adult ECMO circuits were tested for sterility for 56 days between September-December 2020. Circuits were assembled and primed in a perfusion lab in Chicago, IL. Six were stored in a sterile environment and six in a non-sterile environment, with three circuits primed using normal saline (NaCl) and three with Plasmalyte-A for each environment. Samples were collected on days 0, 3, 7, 14, 28, 42, and 56 in anaerobic bottle cultures testing for potential pathogen growth, such as Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. Samples obtained from the 12 primed ECMO circuits demonstrated no microbial growth of S. aureus, P. aeruginosa, and E. coli in the bottle cultures. Similarly, there was no difference in the circuit sterility based on the storage environment (sterile vs nonsterile) or priming solution (NaCl vs Plasmalyte-A). Our findings showed that ECMO circuits can be primed for 56 days without evidence of the specified bacterial growth. Furthermore, the storage conditions and the prime utilized did not affect the sterility of the primed ECMO circuits.

Microbiological performance and adherence in blood culture protocols: The role of a second anaerobic bottle

BackgroundBacteremia represents high rates of morbidity and mortality, especially in developing countries, highlighting the need for a diagnostic method that allows prompt and appropriate patient treatment. This study compared microbiological performance and adherence of two blood culture protocols for the diagnosis of bacteremia. MethodsQuasi-experimental study conducted between June 2022 and February 2023. Two blood culture protocols were evaluated. Protocol 1 included two aerobic bottles and one anaerobic bottle. Protocol 2 included two aerobic and two anaerobic bottles. Protocols were analyzed in three phases: evaluation of protocol 1 (Phase 1); evaluation of protocol 1 plus educational activities for healthcare staff (Phase 2) and evaluation of protocol 2 (Phase 3). Results342 patients and 1155 blood culture bottles (732 aerobic and 423 anaerobic) were included. Positivity was 17.6 %, 22.8 % and 19.4 % in phases 1, 2 and 3, respectively. Among patients with bacteremia, 84.5 % had positive anaerobic bottles, with 9.9 % showing growth only in this bottle. The contamination rates were 1.9 %, 0.3 %, and 0.8 % for each phase, mainly in aerobic bottles. Median positivity time was 11 h for both bottes aerobic and anaerobic. Overall nursing adherence increased from 13.1 % in Phase 1, 25.9 % in Phase 2, and 28.1 % in Phase 3 (p = 0.009). ConclusionsThe findings indicate that adding a second anaerobic bottle does not enhance blood culture positivity. Rather than increasing bottle quantity, staff training might be a more effective approach to optimize results.

Evaluation of BACTEC™ FX and BacT/Alert™ Automated Blood Culture Systems for Detection of Clinically Relevant Bacterial and Yeast Species

Background: Bloodstream infections (BSI) are a significant health concern, necessitating effective diagnostic tools. This study evaluates the performance of the BD Bactec FX system in comparison to the established BacT/Alert system, examining mean time till detection (TTD) across various blood culture bottles.Methods: The study involves 148 blood culture bottles, inoculated with representative bacterial ATCC strains (n= 15) and 04 yeast isolates. Parallel testing is conducted by seeding 10–30 colony-forming units (CFU) in duplicate in both BD Bactec FX and BacT/Alert systems. TTD is assessed across aerobic, anaerobic, and pediatric bottles for diverse microbial species.Results: All 148 bottles tested in parallel show positive signals in both systems. BD Bactec FX demonstrates significantly shorter TTD for adult-seeded cultures in aerobic and anaerobic bottles compared to BacT/Alert Similarly, pediatric bottles with BD Bactec FX exhibit a shorter TTD compared to BacT/Alert. The statistical significance of TTD, 95% confidence intervals (CI), and p-values is evident for Bactec FX aerobic, anaerobic, and pediatric bottles across tested organisms. Notable examples of faster TTD include; Bacteroides ovatus (16.6h by Bactec FX vs. 70.5h by Bact/Alert 3D), Stenotrophomonas maltophilia (33.7h vs. 72.5h), and Streptococcus pyogenes (8.1h vs. 11.9h).Conclusions: In conclusion, the study’s findings demonstrate that BD Bactec FX surpasses BacT/Alert in prompt microbial detection, showcasing potential for early identification of bacteremia and fungemia. Faster TTD implies the potential to initiate timely antimicrobial treatment, thereby reducing patient morbidity and mortality. However, exceptions in certain microbial species highlight the need for comprehensive clinical validation to establish the broad applicability of these findings.Keywords: Blood Culture System; Recovery Rate; Time to detection; Blood stream infection; BTA3D; Bactec FX

Microbiological screening of corneas stored in organ culture medium at Lions Eye Bank of Western Australia from 2011 to 2022.

The purpose of this study was to analyse the contamination rate of corneal samples stored in OCM at Lions Eye Bank of Western Australia over a 12-year period. All OCM samples used to preserve corneas from 2011 to 2022 (inclusive) underwent microbiological testing. Samples were collected into aerobic and anaerobic culture bottles on day 3-5 of corneal preservation and 24h after transfer to thinning medium. Samples were tested for 7days using the BACTEC FX system. Corneas remained in quarantine until clearance was obtained. From 2011 to 2022, 3009 corneas were retrieved and 2756 corneas were stored in OCM. Thirty one (1.1%) positive samples were reported, with 20 growths of bacterial origin and 11 fungal. Microbial contamination was mostly identified on day 1 of culture (77.5%). Donors of contaminated samples had a mean age of 55years, with 17 male and 14 female donors. The highest incidence of contamination came from donors whose cause of death was cancer. Death to enucleation times of contaminated samples ranged from 3.5 to 25.5h (mean = 13.5 ± 7.3) and death to preservation time ranged from 4.1 to 27.5h (mean = 14.8 ± 7.2). These did not significantly differ from the average time from death to enucleation (mean = 13.9 ± 3) and death to preservation (mean = 16.3 ± 4.2) of non-contaminated samples. Microbiological screening of corneas stored in OCM at LEBWA showed a very low rate of positive cultures with no predictive donor characteristics.

Identification of novel bacterial species in the blood of patients with neonatal sepsis

BackgroundThe clinical diagnosis of neonatal sepsis remains difficult because of various challenges, such as culturing the bacteria and avoiding contamination. Therefore, this study aimed to identify bacterial pathogens in patients with clinically diagnosed neonatal sepsis by next-generation sequencing (NGS).MethodsHigh-throughput NGS and traditional culture identification were performed by comparing samples from newborns with neonatal sepsis with healthy control infants. All blood samples were separately inoculated into anaerobic and aerobic bottles and incubated for 7 days at 37 °C, the positive specimens were then identified. Novel bacteria identified through high-throughput NGS were analysed using polymerase chain reaction (PCR), PCR products were verified by Sanger sequencing. Wilcoxon rank-sum and chi-square tests were performed to assess the significance of differences in species abundance between groups. Subjects were clinically diagnosed and hospitalized at the Pediatrics Department of Shenzhen Seventh People’s Hospital and Pediatrics Department of the Longhua Branch of Shenzhen People’s Hospital. Experiments were performed at the Shenzhen Seventh People’s Hospital. The experimental group comprised 45 newborns clinically diagnosed with neonatal sepsis (age: 0–28 days; 28 males, 17 females). Fifteen normal newborns aged 0–28 days (7 males, 8 females) were included as the control group.ResultsHigh-throughput NGS showed a positivity rate of 44% (20/45) for bacteria in patients clinically diagnosed with neonatal sepsis, whereas traditional bacterial culture identification showed a positivity rate of 0% (0/45). The four main bacterial species identified were Anoxybacillus kestanbolensis, Geobacillus vulcani, Klebsiella oxytoca, and Acinetobacter guillouiae.ConclusionsA. kestanbolensis, G. vulcani, K. oxytoca, and A. guillouiae, newly discovered bacteria in patients with neonatal sepsis, were identified with high-throughput NGS. Which may result from maternal intrauterine infection or birth-canal infection and have a high clinical-cure rate. Owing to a lack of methods to culture these bacteria, their role in neonatal sepsis remains unclear. A definite diagnosis cannot rely solely on bacterial culture identification for patients with a suspected diagnosis and clinical diagnosis of neonatal sepsis and should involve other effective diagnostic techniques.

Utility of Anaerobic Blood Cultures in Neonatal Sepsis Evaluation.

Clinicians variably obtain anaerobic blood cultures as part of sepsis evaluations in the neonatal intensive care unit (NICU). Our objective was to determine if anaerobic blood culture bottles yielded clinically relevant information by either recovering pathogens exclusively or more rapidly than the concurrently obtained aerobic culture bottle in the NICU. A retrospective cohort study of blood cultures obtained from infants admitted to the NICU from August 01, 2015 to August 31, 2023. Standard practice was to inoculate 2 mL of blood divided equally between an aerobic and an anaerobic culture bottle. We analyzed positive blood cultures where both aerobic and anaerobic bottles were obtained and compared pathogen recovery and time to positivity between the bottles. During the study period, 4599 blood cultures were obtained from 3665 infants, and 265 (5.8%) were positive. Of these, 182 cultures were sent as aerobic-anaerobic pairs and recovered pathogenic organisms. Organisms were recovered exclusively from the anaerobic bottle in 32 (17.6%) cultures. Three organisms were obligate anaerobes; the rest were facultative anaerobes including Coagulase-negative staphylococci (40.6%), Escherichia coli (15.6%), and Staphylococcus aureus (15.6%). Cultures with exclusive recovery in the anaerobic bottle were more frequently obtained ≤3 days after birth, compared to other cultures (31.3% vs 15.3%, P = .03). When both bottles recovered the pathogen (n = 113), the anaerobic bottle had a shorter time to positivity in 76 (67.3%) cultures. Including anaerobic culture bottles could lead to the identification of pathogens not recovered in the aerobic bottle, as well as earlier identification of pathogens.

Effects of yak rumen anaerobic fungus Orpinomyces sp. YF3 fermented on in vitro wheat straw fermentation and microbial communities in dairy goat rumen fluid, with and without fungal flora.

Rumen fungi play an essential role in the breakdown of dietary fibrous components, facilitating the provision of nutrients and energy to the host animals. This study investigated the fermentation characteristics and effects on rumen microbiota of yak rumen anaerobic fungus Orpinomyces sp. YF3 in goat rumen fluid, both with and without fungal flora, utilizing anaerobic fermentation bottles. Crushed and air-dried wheat straw served as the fermentation substrate, and cycloheximide was used to eradicate microorganisms from the rumen fluid of dairy goats. The experiment compromised four treatment groups (2×2 factorial design): control (C); yak fungus group (CF, Orpinomyces sp. YF3); goat fungi eliminated group (CA, antibiotic: 0.25 mg/mL cycloheximide); goat fungi eliminated+yak fungus group (CAF). Each treatment had six replicates. Fermentation characteristics and microbial composition of the fermentation media were analyzed using one-way analysis of variance and high-throughput sequencing technology. The findings revealed that in the Orpinomyces sp. YF3 addition group (CF and CAF groups), there were significant increases in ammonia nitrogen concentration by 70%, total volatile fatty acids (VFA) by 53%, as well as acetate, isobutyrate, and valerate concentrations, and the ratio of acetate to propionate (p < 0.05), while the propionate proportion declined by 13%, alongside a reduction of butyrate concentration (p < 0.05). Similarly, in the CF and CAF groups, there were a notable increase in the relative abundance of Bacteroidota, Synergistota, Desulfobacterota, Actinobacteria, and Fusobacteriota, alongside a decrease in the relative abundance of Fibrobacterota and Proteobacteria (p < 0.05). Bacteria exhibiting increased relative abundance were positively correlated with the activity of carboxymethyl cellulase and avicelase, total VFA concentration, and acetate proportion, while showing a negatively correlation with propionate proportion. In conclusion, supplementing rumen fermentation media with yak rumen anaerobic fungus Orpinomyces sp. YF3 led to an increase in bacteria associated with fibre degradation and acetic acid production, a decrease in propionate-producing bacteria, enhanced the activity of plant cell wall degrading enzymes, and promoted cellulose degradation, ultimately elevating total VAF concentration and acetate proportion. This presents a novel approach to enhance roughage utilization in ruminants.

Optimized decision algorithm for the microbiological diagnosis of osteoarticular infections in adults using synovial fluid samples: a prospective study in two French hospitals including 423 samples of synovial fluid.

No consensus exists about the techniques to use for microbiological diagnosis of bone and joint infections (BJIs). The objective herein was to define an algorithm to optimize BJI diagnosis in adults using various bacteriological methods on synovial fluid samples. This prospective multi-center study included 423 synovial fluids collected from adult patients with suspected BJIs. Culture (using five solid media, an enrichment broth, and blood culture bottles), universal 16S rRNA PCR followed by Sanger sequencing, and seven specific bacterial PCRs were systematically performed. Combinations of methods were compared to arrive at the optimized algorithm. Among 423 synovial fluids, 242 infections were diagnosed (57.2 %): 213 mono- and 29 poly-microbial for a total of 284 bacteria (staphylococci at 54.6 %, streptococci-enterococci at 16.5 %, Gram-negative bacilli at 15.5 %, anaerobic species at 8.8 %). Comparing culture techniques, blood culture bottles had the highest sensitivity (67.6 % for pediatric and 63.9 % for anaerobic bottles) but are not sufficient alone and require being combined with solid media. The 16S rDNA PCR detected only 52.3 % of the bacteria, whereas specific PCRs had a higher sensitivity (Staphylococcus spp. at 66.2 %, S.aureus at 85.2 %, Streptococcus spp. at 91.2 %). Based on these results, an algorithm was proposed associating three solid media; inoculation into blood culture bottles; and 16S, Staphylococcus spp., and Streptococcus spp. PCRs, which would have detected 90.5 % of bacteria in the present cohort versus 79.2 % using all culture techniques on synovial fluid. This prospective study shows that a combination of culture and molecular methods on synovial fluids allows the optimization of bacterial detection.

Bacterial Contamination of Platelet Products.

Transfusion of bacterially contaminated platelets, although rare, is still a major cause of mortality and morbidity despite the introduction of many methods to limit this over the past 20 years. The methods used include improved donor skin disinfection, diversion of the first part of donations, use of apheresis platelet units rather than whole-blood derived pools, primary and secondary testing by culture or rapid test, and use of pathogen reduction. Primary culture has been in use the US since 2004, using culture 24 h after collection of volumes of 4-8 mL from apheresis collections and whole-blood derived pools inoculated into aerobic culture bottles, with limited use of secondary testing by culture or rapid test to extend shelf-life from 5 to 7 days. Primary culture was introduced in the UK in 2011 using a "large-volume, delayed sampling" (LVDS) protocol requiring culture 36-48 h after collection of volumes of 16 mL from split apheresis units and whole-blood derived pools, inoculated into aerobic and anaerobic culture bottles (8 mL each), with a shelf-life of 7 days. Pathogen reduction using amotosalen has been in use in Europe since 2002, and was approved for use in the US in 2014. In the US, recent FDA guidance, effective October 2021, recommended several strategies to limit bacterial contamination of platelet products, including pathogen reduction, variants of the UK LVDS method and several two-step strategies, with shelf-life ranging from 3 to 7 days. The issues associated with bacterial contamination and these strategies are discussed in this review.

Performance evaluation of 4-day versus 5-day blood cultures using the BD BACTEC FX system

Background: Blood culture (BC) systems have evolved to increase sensitivity and reduce turnaround times. This study compared the performance of a 4-day versus a 5-day BC incubation period using the BD BACTEC™ FX (Becton, Dickinson and Company, USA). Methods: A total of 37,379 consecutive sets of BC were evaluated over a 3-month period in a 2,700-bed tertiary care hospital. Positive BC results were reviewed to assess time-to-positivity (TTP) and species identification of the isolates. The BCs were performed in pairs of vials, utilizing either BD BACTEC Plus Aerobic/F or Peds Plus/F with BD BACTEC Lytic Anaerobic media. Results: A total of 14,899 episodes, averaging 2.51 sets per episode, were analyzed. Of these, 1,398 (9.38%) were positive, yielding 1,465 isolates. TTP (hours) were &lt; 12 in 48.87%, 12-24 in 31.40%, 24-48 in 13.38%, 48-72 in 3.28%, 72-96 in 1.43%, and &gt;96 in 1.64%. The two most prevalent organisms, Escherichia coli and Klebsiella spp. were detected within 12 hours in 88.75% and 78.90%, respectively. The respective median TTP (T50) values for E. coli, Klebsiella spp., Enterococcus faecalis/E. faecium, and Staphylococcus aureus were 9.24, 9.60, 13.75, and 14.20. T50 values for these species were significantly shorter in anaerobic bottles than in aerobic bottles. Of 24 BCs with TTP &gt; 96, only 4 containing anaerobic bacteria or molds were f irst detected after 96 hours. Conclusion: A 4-day incubation has demonstrated excellent sensitivity. However, a 5-day incubation may be beneficial for hospitals caring for patients at high risk for infections with slow-growing fungi or fastidious bacteria.

Comparison of new and old BacT/ALERT aerobic bottles for detection of Candida species.

A new version of aerobic blood culture media has been developed for the BacT/ALERT (bioMérieux) blood culture system. We evaluated the time to detection and yeast cell counts in positive blood cultures for each Candida spp. according to changes in media. Isolates from defibrinated horse blood were inoculated into three types of bottles: the old version of aerobic bottle, new version of aerobic bottle, and anaerobic bottle. All bottles were incubated in the BacT/ALERT Virtuo blood culture system. The time to detection was monitored for each bottle, and yeast cell counts were performed immediately after testing positive, determined via the plate count method. Clinical retrospective data of the candidemia samples before and after aerobic bottle change also were analyzed. The median time to detection was 52.47 hours in the old aerobic bottles versus 19.92 hours in the new aerobic bottles (P < 0.001) for Candida glabrata, and standard and clinical strains showed similar results. C. albicans (27.6 to 24.95 hours) and C. guilliermondii (28.92 to 26.9 hours) had shorter time to detection. However, C. auris (25.43 to 28.25 hours) had a longer time to detection in the new aerobic bottle. The retrospective clinical analysis showed a significant decrease in time to detection (45.0 to 19.4 hours) for C. glabrata, which is consistent with our simulated study result for C. glabrata. As a result of analysis including all blood specimens, C. tropicalis showed a significant delay in time to detection in new aerobic bottles. In an analysis limited to peripheral blood specimens, the time to detection of C. parapsilosis was longer in new aerobic bottles than in old aerobic bottles. Most Candida species did not show remarkable TTD differences, but TTD of C. glabrata was markedly reduced in the New FA Plus bottle. The reduction of time to detection enables faster detection and therapeutic approach for C. glabrata infections.

172. Impact of a quality improvement re-training program on the blood culture volume, yield and contamination rates; experience from South India

Abstract Background Blood cultures help to detect causative agents in bacteremia and are important for targeted therapy. Adequate blood culture volume is considered as 8-10ml in each bottle and at least 2 aerobic and 2 anaerobic blood culture bottles are recommended. Inadequate blood culture fills are common and may affect yield. Figure 1 Methods The study was done in our healthcare facility-KIMSHEALTH, Trivandrum, India. Pre-intervention period was between May 2021 and June 2021; post-intervention was between May 2022 and June 2022. The intervention was through plan/do/study/act (PDSA) cycles and included multiple training sessions to the phlebotomy team regarding adequate fill volume which is 8-10 ml in each blood culture bottle. Training for maintaining asepsis during collection was reinforced. All blood culture samples received in the hospital microbiology laboratory during the above period were included in this study and compared between the pre-intervention and post-intervention period. Pediatric blood culture samples were excluded. Results Blood culture samples studied during pre and post intervention period were 1428 and 1716 respectively. In the pre-intervention phase adequate fill volume was noted in 30.81% bottles (figure 1), while in the post intervention adequate fill volume was noted in 68.06% bottles (p &amp;lt; 0.01). Blood culture contamination rate was 3.01% in the pre-intervention period which improved to 2% in the post-intervention period and the difference was significant (p&amp;lt; 0.01). Blood culture yield of a pathogenic organism did not show significant difference after the intervention (11.2% in pre-intervention group Vs 11.45% in post intervention group). Conclusion In this study, an improvement was noted in the blood culture fills by 38% and contamination rate reduced by 1%. Though there was no significant increase in blood culture yield with intervention, the reduction in contamination was noted which will reduce unnecessary antibiotic initiation. Disclosures All Authors: No reported disclosures

587. Research and Development of Image Recognition AI to Estimate Bacterial Species using Gram Stain Findings in Aerobic and Anaerobic Blood Culture Bottle

Abstract Background Background In the highly fatal infectious disease of bacteremia, the ability to select antimicrobial agents at an earlier time is critical not only for early patient recovery, but also for reducing the development of resistant organisms. Gram staining of blood cultures is a useful test for early selection of antimicrobial agents, but it requires some skill in deciphering. Therefore, we developed an image analysis system for Gram stain images of blood cultures and have verified whether it is possible to estimate the bacterial species to facilitate the initial selection of antimicrobial agents, regardless of the level of proficiency. Methods Method Slides and bacterial species-identification information from an anonymized Gram stain registry at two medical institutions, National Center for Global health and Medicine (NCGM) and Kobe University Hospital (KUH), were used for the study. 1113 cases of aerobic bottles and 1060 cases of anaerobic bottles were included. Mock-specimens were used for the rare bacteria. A total of 23,947 images were generated by capturing the observation field of view of an optical microscope with a smartphone. Table 1 shows the bacterial classification of aerobic bottles and Table 2 shows the bacterial classification of anaerobic bottles. The data were divided into training data and test data at a ratio of 8 to 2 for each category.Table 1.A total of 16 categories of bacteria were used for classification of aerobic bottlesTable 2.A total of 16 categories of bacteria were used for classification of anaerobic bottles Results The macro-average recall (sensitivity) and accuracy of the test data were 64% and 82% for aerobic bottles and 68% and 86% for anaerobic bottles, respectively. The results showed the possibility of classifying the performing bacterial species with an accuracy of over 70% using only Gram stained images via image recognition AI. For aerobic bottles in particular, the seven categories of GNR, GNC, GPR, GPC, yeast, No organism, and multiple bacteria are predicted to have 97% accuracy, and 90% macro average recall. Conclusion The research has shown that it is possible to classify bacterial species to a certain extent only by observing the Gram stain images. Our goal is to narrow down and focus more on the clinically-meaningful bacterial species, and to improve the accuracy of our classification. Also, we are planning to make comparisons with specialists in the future. Disclosures Kei Yamamoto, MD, Canon medical systems: Grant/Research Support|CarbGeM: Grant/Research Support|CarbGeM: 7090302|Fujirebio: Grant/Research Support|Sanyo Chemical Industries: Grant/Research Support|VisGene: Grant/Research Support Goh Ohji, MD, PhD, DTMH, CarbGeM: Advisor/Consultant

Clinical usefulness of anaerobic blood culture in pediatric patients with bacteremia

ObjectivesThe purpose of this study is to evaluate the usefulness of anaerobic blood culture in pediatric patients by comparing the detection rate and distribution of bacteria between aerobic and anaerobic blood culture bottles. MethodsA retrospective analysis was conducted on 11,664 blood cultures obtained from children under the age of 14 between January 2013 and June 2020. The positive rate of total, aerobic, and anaerobic blood culture, as well as the species distribution of each blood culture bottle, were investigated. ResultsThe positive rate of blood culture was 2.4 % (N = 281). Among them, 67 (23.8 %), 85 (30.3 %) and 129 (45.9 %) organisms were grown in only aerobic, only anaerobic, and both blood culture bottles, respectively. Gram-positive cocci were cultured on both, only aerobic, and only anaerobic blood culture bottles in proportions of 46.4 %, 23.4 %, and 30.2 %, respectively. Gram-negative bacilli were cultured on both, only aerobic, and only anaerobic blood culture bottles in proportions of 58.5 %, 12,3 %, and 29.2 %, respectively. Gram-positive bacilli grew best in aerobic bottle only. There were seven strains of obligate anaerobes. ConclusionBecause many facultative anaerobic bacteria are recognized primarily from anaerobic blood culture bottles, combining aerobic and anaerobic blood culture bottles might be beneficial in pediatric patients with suspected blood stream infection.

Limiting acetoin generation during 2,3-butanediol fermentation with Paenibacillus polymyxa using lignocellulosic hydrolysates

Recent decarbonization efforts have led to interests in producing more bio-based chemicals. One attractive compound produced biochemically is the platform chemical known as 2,3-butanediol (2,3-BDO). In this work a mild alkaline pretreatment using sodium carbonate was performed on corn stover (CS) and switchgrass (SG) to generate hydrolysates for fermentation with the 2,3-BDO producer bacteria strain Paenibacillius polymyxa. Enzymatic hydrolysis performed on the pretreated CS and SG produced theoretical sugar yields of 80 % and 95 % for glucose and xylose, respectively. Fermentations with P. polymxya conducted in anaerobic bottles produced 2,3-BDO reaching concentrations ranging from 14 to 18 g/L with negligible conversion into acetoin. Bioreactor fermentations using the hydrolysate media generated up to 43 g/L and 34 g/L of 2,3-BDO from pretreated CS and SG, respectively, within 24 h of fermentation. However, 2,3-BDO product output was reduced by 40–50 % over the remainder of the fermentation due to conversion into acetoin caused by glucose depletion.

Nutrient supplementation of culture media improves the detection of Cutibacterium acnes in platelet components by an automated culture system.

Platelet concentrates (PCs) contaminated with Cutibacterium acnes are often transfused prior to detection by the BACT/ALERT system. Though C. acnes is implicated in mild transfusion reactions, delayed clinical effects are unknown. This study assessed the ability to enhance C. acnes detection by supplementing culture media with Tween 80 (T80, an oleic acid source) and a commercial nutrient supplement. Anaerobic culture bottles (BPN) were supplemented with T80 or oleic acid. T80-supplemented BPN bottles were inoculated with four C. acnes isolates (10 or 100 colony-forming units [CFU]/bottle) or other transfusion-relevant bacteria (10 CFU/bottle). Samples of plasma containing SSP+ (platelet additive solution [PAS]) (PAS-plasma) at different concentrations, plasma-PCs and PAS-PCs, spiked with two C. acnes isolates (10 CFU/bottle), were inoculated into T80-supplemented BPN bottles. Furthermore, plasma-PCs were spiked with C. acnes and tested in BPN bottles supplemented with the BD Difco Supplement VX (BDVx). Bottles were incubated in the BACT/ALERT system and times to detection (TtoD) were compared (N = 3). A reduction in TtoD of ≤3.5 days was observed for C. acnes in T80-supplemented BPN, while other species did not show the same effect. However, false positives were observed when T80-supplemented BPN was inoculatedwith PAS-plasma (except for 70% PAS:30% plasma), plasma-PCs or PAS-PCs. Oleic acid supplementation also resulted in false positives. Interestingly, BDVx-supplemented BPN reduced the TtoD of C. acnes in PCs by ≤1.2 days (p < 0.05), with no false-positive results. BDVx supplementation for detection of C. acnes from PCs could result in timely unit retrieval, preventing the transfusion of contaminated products. In clinical settings, T80 supplementation could significantly enhance C. acnes detection from non-blood-derived samples.

Dual-Temperature Microbiological Control of Cellular Products: A Potential Impact for Bacterial Screening of Platelet Concentrates?

An experimental study by the Paul-Ehrlich Institute (PEI) demonstrated that temperatures between 35 and 37 °C are too high for the growth of some bacterial strains (e.g., Pseudomonas fluorescens), leading to false negative results. Thus, the question of whether it is necessary to adapt incubation temperatures for the microbiological control of blood products, especially platelet concentrates (PCs), to enhance safety and regulatory compliance has arisen. In order to further elucidate this issue, the growth capability of different bacterial strains of interest in PCs and the detection efficacy of cultivation of these at different incubation temperatures must be taken into account. Therefore, we inoculated PCs with 46 different strains (3-6 PCs from different donors per strain) from different origins (PC isolates, reference strains) and stored PCs at 20-22 °C under constant agitation. On day three of storage, the inoculated PCs were sampled; aerobic and anaerobic culture bottles (BacT/Alert AST/NST) were each inoculated with 5 mL of sample, and culture bottles were incubated at 25 and 35 °C using the automated BacT/Alert Dual-temperature system. Bacterial proliferation was enumerated using a colony-forming assay. All strains of Enterobacteriacae (n = 5), Staphy-lococcus spp. (n = 11), Streptococcus spp. (n = 5), and Bacillus spp. (n = 4) and most Pseudomonas aeruginosa strains (4 of 5) tested showed the capability to grow in most inoculated PCs, revealing a faster time to detection (TTD) at an incubation temperature of 35 °C. The tested Pseudomonas putida (n = 3) strains showed a noticeably reduced capability to grow in PCs. Nonetheless, those with a notable growth capability revealed a faster TTD at an incubation temperature of 35 °C. Only one of the four Pseudomonas fluorescens strains tested (strain ATCC 13525) was able to grow in PCs, showing a faster TTD at an incubation temperature of 25 °C but also detection at 35 °C. The commonly detected bacteria involved in the bacterial contamination of PCs showed a superior TTD at 35 °C incubation. Only one P. fluorescens strain showed superior growth at 25 °C; however, the microbiological control at 35 °C did not fail to identify this contamination. In conclusion, the use of PC screening using a dual-temperature setting for microbiological control is presently not justified according to the observed kinetics.

Validation of the Accuracy of Automatic Measurement of Blood Volume in Culture Bottles for Blood Culture.

Several manufacturers have developed systems that automatically measure the amount of blood in culture bottles. We compared the volumes measured automatically by the Virtuo instrument (bioMérieux, France) (height-based volumes) and those calculated by weighing the bottles. In all, 150 pairs of aerobic and anaerobic blood culture bottles (BacT/ALERT FA/FN Plus, bioMérieux) were randomly selected over two periods to compare the height- and weight-based volumes and analyze the effect of foam. We also estimated the limit of detection (LOD) and the cut-off value for 5 mL equine blood. The mean height-based volume was approximately 0.67 mL greater than the weight-based volume, particularly in anaerobic culture bottles. Foam did not have a significant effect. The LOD for the automatic height-based volume of equine blood was 0.2-0.4 mL. The 5 mL cut-off was 4-4.2 mL. Therefore, when reporting or monitoring blood volume within culture bottles in the laboratory, these performance characteristics should be adequately considered.

Microbial characteristics and potential mechanisms of souring control for a hypersaline oil reservoir

The production of hydrogen sulfide (H2S) had caused huge economic losses and security risks to oil fields, which had attracted the attention of many researchers. Studies focused on hypersaline oil reservoirs were scarce. Here, we simulated H2S inhibition by adding NO3 - and NO2 - in anaerobic bottles and up-flow sand-packed bioreactor in the laboratory using water samples from injection wells in the Jianghan reservoir. The microbial characteristics, pH, salinity, sulfate, sulfide, NO3 - and NO2 - were measured at every stage. The content of H2S and elemental sulfur (S0) gradually reduced to 0 mg/L with injected 400 mg/L of NO3 - and 300 mg/L of NO2 -. Halophilic nitrate-reducing bacteria (NRBs) related to Halomonas and Arcobacter could suppress the activity of sulfate-reducing bacteria (SRBs) through the accumulation of nitrite toxicity and the competition of organic substrates with SRBs. When SRBs activity could not be suppressed, the sulfur-oxidizing bacteria related to Sulfurimonas, Thiomicrospira and Acrobacter were accumulated for souring control through microbial sulfur cycle of H2S production and oxidation. Salinity could be a key parameter affecting microbial development and interactions. This study provides a fundamental data to souring control in the hypersaline oil reservoir by reinjection produced water amended with NO3 - and NO2 -.

Increased frequency of false-positive bacterial detection after implementation of new guidelines for large-volume delayed sampling of platelets.

The updated guidance for improving bacterial detection (BD) of platelets has included the implementation of large-volume delayed sampling (LVDS) with the addition of anaerobic culture bottles (BPNs) and sampling of each platelet split product. The frequency of BD was reviewed during this LVDS time period in comparison with pre-LVDS and the Post-Approval Surveillance Study of Platelet Outcomes, Release Tested (PASSPORT) study (when BPNs were last used). There was more than a twofold increase in bottles inoculated per collection during LVDS, with an almost fivefold increase in sample volume collected. During LVDS, the concordance of split products within an initial reactive collection was only 8.7%. There was no difference in LVDS aerobic culture bottle (BPA) true positives (TPs), but there was a significant increase in LVDS false positives (FPs), p < .0001, compared to both PASSPORT and pre-LVDS, respectively. There was an increase in BPN TPs during LVDS (p < .05 compared to PASSPORT), with predominance of Cutibacter acnes (C. acnes), noted exclusively in BPN, and accounting for more than two-fifths of all organisms detected. Time to alarm during LVDS for TPs had two peaks with one due to C. acnes at 96 h compared to 17 h for non-C. acnes. The high FP frequency, along with low clinical significance of TPs found in BPNs, has led to the needless discard of inventory, as the utility of BPNs in BD for platelets is yet to be established and may require much larger studies.