Implications of Ad-Hoc Molecular Typing for Infection Control Measures in a Multi-cluster, Multi-phenotypic Serratia marcescens Outbreak in a Neonatal Intensive Care Unit.

INTRODUCTION: Serratia marcescens is known to cause outbreaks in Neonatal Intensive Care Units (NICU). Traditionally epidemiological data, antimicrobial resistance patterns and epidemiological typing are being used to guide infection prevention methods. Whole genome sequencing (WGS) applications such as core-genome multi-locus sequence typing (cgMLST) applied during an outbreak would potentially give more detailed information on the source and spread of bacteria enabling more efficient control measures. CgMLST was used during an S. marcescens outbreak at a NICU. METHODS: Neonates admitted to the NICU of the Leiden University Medical Center (LUMC) during an outbreak between September 2023-January 2024, with S. marcescens being cultured were included. Environmental samples were taken to search for a common source, antibiotic susceptibility testing was performed, and antimicrobial resistances genes were analysed. RESULTS: S. marcescens strains from 17 of the 20 positive patients were available for molecular typing. The cgMLST scheme revealed five different complex types consisting of four separate clusters. Multiple clusters made an unidentified persistent environmental source as cause of the outbreak less likely, leading to a quick downscaling of infection prevention measures. Differences were shown in aminoglycoside resistance patterns of isolates within the same complex types and patients. DISCUSSION: The use of ad-hoc cgMLST provided timely data for rational decision making during an S. marcescens outbreak at the NICU. Antibiotic phenotyping alone was found not to be suitable for studying clonal spread during this outbreak with S. marcescens.

Antimicrobial susceptibility profile of clinically relevant Bacteroides, Phocaeicola, Parabacteroides and Prevotella species, isolated by eight laboratories in the Netherlands.

OBJECTIVES: Recently, reports on antimicrobial-resistant Bacteroides and Prevotella isolates have increased in the Netherlands. This urged the need for a surveillance study on the antimicrobial susceptibility profile of Bacteroides, Phocaeicola, Parabacteroides and Prevotella isolates consecutively isolated from human clinical specimens at eight different Dutch laboratories. METHODS: Each laboratory collected 20-25 Bacteroides (including Phocaeicola and Parabacteroides) and 10-15 Prevotella isolates for 3 months. At the national reference laboratory, the MICs of amoxicillin, amoxicillin/clavulanic acid, piperacillin/tazobactam, meropenem, imipenem, metronidazole, clindamycin, tetracycline and moxifloxacin were determined using agar dilution. Isolates with a high MIC of metronidazole or a carbapenem, or harbouring cfiA, were subjected to WGS. RESULTS: Bacteroides thetaiotaomicron/faecis isolates had the highest MIC90 values, whereas Bacteroides fragilis had the lowest MIC90 values for amoxicillin/clavulanic acid, piperacillin/tazobactam, meropenem, imipenem and moxifloxacin. The antimicrobial profiles of the different Prevotella species were similar, except for amoxicillin, for which the MIC50 ranged from 0.125 to 16 mg/L for Prevotella bivia and Prevotella buccae, respectively. Three isolates with high metronidazole MICs were sequenced, of which one Bacteroides thetaiotaomicron isolate harboured a plasmid-located nimE gene and a Prevotella melaninogenica isolate harboured a nimA gene chromosomally.Five Bacteroides isolates harboured a cfiA gene and three had an IS element upstream, resulting in high MICs of carbapenems. The other two isolates harboured no IS element upstream of the cfiA gene and had low MICs of carbapenems. CONCLUSIONS: Variations in resistance between species were observed. To combat emerging resistance in anaerobes, monitoring resistance and conducting surveillance are essential.

Characterization of mobile genetic elements in multidrug-resistant Bacteroides fragilis isolates from different hospitals in the Netherlands.

OBJECTIVES: Five human clinical multidrug-resistant (MDR) Bacteroides fragilis isolates, including resistance to meropenem and metronidazole, were recovered at different hospitals in the Netherlands between 2014 and 2020 and sent to the anaerobic reference laboratory for full characterization. METHODS: Isolates were recovered from a variety of clinical specimens from patients with unrelated backgrounds. Long- and short-read sequencing was performed, followed by a hybrid assembly to study the presence of mobile genetic elements (MGEs) and antimicrobial resistance genes (ARGs). RESULTS: A cfxA gene was present on a transposon (Tn) similar to Tn4555 in two isolates. In two isolates a novel Tn was present with the cfxA gene. Four isolates harbored a nimE gene, located on a pBFS01_2 plasmid. One isolate contained a novel plasmid carrying a nimA gene with IS1168. The tetQ gene was present on novel conjugative transposons (CTns) belonging to the CTnDOT family. Two isolates harbored a novel plasmid with tetQ. Other ARGs in these isolates, but not on an MGE, were: cfiA, ermF, mef(EN2), and sul2. ARGs harboured differed between isolates and corresponded with the observed phenotypic resistance. CONCLUSIONS: Novel CTns, Tns, and plasmids were encountered in the five MDR B. fragilis isolates, complementing our knowledge on MDR and horizontal gene transfer in anaerobic bacteria.

Toxinotype A Clostridium perfringens causing septicaemia with intravascular haemolysis: two cases and review of the literature.

BACKGROUND: Septicaemia with intravascular haemolysis is a rare, but often fatal, presentation of Clostridium perfringens infection. C. perfringens is a Gram-positive, anaerobic bacterium that can produce multiple toxins. Toxinotyping is not performed regularly. METHODS: This article describes two human cases of C. perfringens infections. Toxinotyping was performed using polymerase chain reaction (PCR). Additionally, a structured review of the literature was performed which searched specifically for cases of C. perfringens infection with haemolysis. RESULTS: Both cases were identified as toxinotype A strains and both cases were fatal. Also, both cases showed marked haemolysis during their clinical course, which is assumed to have played a significant role in their outcome. In total, 83 references were identified describing human C. perfringens infection with haemolysis. Mortality rates have been stable over the last 10 years at 80%. Toxinotyping has been performed in a total of six cases. Of the four cases analysed by PCR, all were identified as toxinotype A. CONCLUSIONS: Haemolytic C. perfringens infections are rare but are fatal in most cases. Toxinotyping is performed rarely. The authors advocate increased use of toxinotyping to gain insight into pathophysiology and more effective interventions.

Ten-Year Follow-Up of Patients Treated with Fecal Microbiota Transplantation for Recurrent Clostridioides difficile Infection from a Randomized Controlled Trial and Review of the Literature.

Fecal microbiota transplantation (FMT) has become a well-established treatment for recurrent Clostridioides difficile infection (rCDI). While short-term outcomes and adverse events relating to FMT have been well documented, there still is a paucity of data with regard to long-term safety. In this report, we describe the long-term follow-up of the prospective cohort of the first randomized controlled trial of FMT for rCDI, and review the existing literature. A total of 34 patients were treated with FMT for rCDI. Seven patients were still alive after a follow-up of more than 10 years and three patients were lost to follow-up. None of the 34 patients had experienced a new-onset autoimmune, gastrointestinal, or malignant disorder during follow-up. We did not find any deterioration or amelioration of pre-existing medical conditions. Furthermore, no deaths directly attributable to FMT could be identified. These findings are in accordance with the data in available literature. In conclusion, no long-term adverse events or complications directly attributable to FMT were found in our prospective cohort. Review of the available literature does not point to long-term risks associated with FMT in this elderly population, provided that carefully screened fecal suspensions are being used. No firm conclusion on the long-term safety of FMT in younger patients could be drawn.

Diagnostic yield of repeat sampling with immunoassay, real-time PCR, and toxigenic culture for the detection of toxigenic Clostridium difficile in an epidemic and a non-epidemic setting.

Current international guidelines lack definite conclusions regarding repeat stool sampling for the detection of toxigenic Clostridium difficile. We assessed the value of repeat sampling and compared the diagnostic yield in an epidemic to a non-epidemic setting. Consecutive fecal samples obtained during two time frames were analyzed using direct stool immunoassay toxin testing (enzyme immunoassay [EIA]), direct stool real-time PCR toxin gene testing, and toxigenic culture. Samples collected within 7 days of the initial sample were considered repeat tests. In the epidemic setting 989 patients were analyzed, and in the non-epidemic setting 1,015. In the epidemic setting 204 patients had two or more specimens included for analysis and in the non-epidemic setting 287 patients. In the epidemic setting 136 samples yielded a positive results, either by EIA or toxigenic culture; of these, 108 were positive according to EIA and 123 according to toxigenic culture. In the first test round 98 (90.7%, 95% CI 85.3 to 96.2), 114 (92.7%, 88.1 to 97.3), and 126 (92.6%, 88.3 to 97.0) positives were detected. Subsequent test rounds yielded 10 (9.3%, 3.8 to 14.7), 9 (7.3%, 2.7 to 11.9), and 10 (7.4%, 3.0 to 11.7) extra positives. In the non-epidemic setting EIA, toxigenic culture and PCR detected 33, 66, and 83 positives. The three tests combined 93 detected positives. In the first test round 30 (90.9%, 81.1 to 100.7), 63 (95.5%, 90.4 to 110.5), 76 (91.6%, 85.6 to 97.5), and 87 (93.5%, 88.6 to 98.5) positives were detected. Subsequent test rounds yielded 3 (9.1%, -0.7 to 18.9), 3 (4.5%, -0.5 to 9.6), 7 (8.4%, 2.5 to 14.4), and 6 (6.5%, 1.5 to 11.4) extra positives. In conclusion, repeat testing resulted in 4.5% to 9.3% extra positives. No significant difference between the settings studied could be demonstrated. Repeat sampling and multimodality testing may be chosen in an outbreak situation to detect all cases, effectively controlling nosocomial spread.

The influence of different types of stent grafts on aneurysm neck dynamics after endovascular aneurysm repair.

OBJECTIVE: Dynamic imaging provides insight into aortic shape changes throughout the cardiac cycle. These changes may be important for proximal aortic stent graft fixation, sealing and durability. The objective of this study is to analyse the influence of different types of stent grafts on dynamic changes of the aneurysm neck. METHODS: Pre- and postoperative electrocardiography (ECG)-gated computed tomographic angiography (CTA) scans were obtained in 30 abdominal aortic aneurysm (AAA) patients, 10 each from three different types of stent grafts (10 Talent, Endurant, and Excluder). Each dynamic CTA dataset consisted of eight reconstructed images over the cardiac cycle. Aortic area and radius changes during the cardiac cycle were determined at two levels: (A) 3 cm above and (B) 1 cm below the lowermost renal artery. Radius changes were measured over 360 axes, and plotted in a polar plot. An ellipse was fitted over the plots to determine radius changes over the major and minor axis for assessment of the asymmetric aspect and most prominent direction of distension. RESULTS: Baseline characteristics did not differ significantly between the three groups. Preoperatively, the aortic area increased significantly (p < 0.001) over the cardiac cycle in all patients at both levels: (A) mean increase 8.3 +/- 4.1% (2.0-17.3%); (B) mean increase 5.9 +/- 4.2% (1.9-12.4%). The postoperative aortic area increase over the cardiac cycle did not differ significantly from preoperative increases: (A) mean increase 9.9 +/- 2.2% (4.4-20.0%); (B) mean increase 7.7 +/- 2.4% (3.8-12.4%). The difference between radius change over the major and minor axis was significant both pre- and postoperatively for all three stent grafts, indicating asymmetric distension. Suprarenal, the distension showed a tendency to right-anterior and infrarenal to left-anterior. The distension and direction of the aortic expansion was preserved after stent grafting. There were no differences between the three types of stent grafts regarding their impact on the aortic distension or direction of this distension. CONCLUSION: The aorta expands significantly and asymmetrically throughout the cardiac cycle. After implantation of abdominal aortic stent grafts, the aortic distension and direction of distension remain equally preserved in all three groups. The three stent graft types studied seem to be able to adapt to the asymmetric dynamic aortic shape changes.

Dynamics of the aorta before and after endovascular aneurysm repair: a systematic review.

OBJECTIVE: An overview of the knowledge of thoracic (TAA), and abdominal aortic aneurysm (AAA) dynamics, before and after endovascular repair, is given. METHODS: Medline, EMBASE and the Cochrane database were searched for relevant articles. After inclusion and exclusion, 25 relevant articles reporting on aneurysm dynamics remained, allowing for comparison. Results provided in the included studies were assumed (statistically) significant if they were larger than the repeatability of the used method. RESULTS: The sample size of dynamic studies is limited and translational studies are missing. Magnetic resonance angiography (MRA) and computed tomographic angiography (CTA) were shown to have lower inter-observer variabilities than ultrasonography (US). The distension of several relevant stent-graft-landing zones during the cardiac cycle in both the abdominal and thoracic aorta are significant (mean diameter change of the AAA neck in the included studies ranged from 0.9 mm to 2.4 mm; mean area change of the thoracic aorta ranged from 4.8% to 12.7% at various levels). This distension remained preserved after stent-graft placement. Preoperatively, the renal arteries displace per heartbeat. Significant movement of the aorta in the anteroposterior (AP) and lateral direction, during the cardiac cycle, was observed. CONCLUSION: The aorta exhibits a wide variety of morphologic changes throughout the cardiac cycle. CTA and MRA are reliable modalities to investigate aortic shape changes during the cardiac cycle. Significant changes per heartbeat are reported in the AAA neck and thoracic aorta. The renal artery displaces per heartbeat. The clinical relevance of dynamic imaging has not been proven yet, but dynamic changes of the aorta have to be taken into account in stent-graft selection and future stent-graft design.

Oversizing of aortic stent grafts for abdominal aneurysm repair: a systematic review of the benefits and risks.

OBJECTIVE: Sizing of aortic endografts is an essential step in successful endovascular treatment of aortic pathology, although consensus regarding the optimal sizing strategy is lacking. Some proximal oversizing is necessary to obtain a seal between the stent graft and the aortic wall and to prevent the graft from migrating, but excessive oversizing might influence the results negatively. In this systematic review, we investigated the current literature to obtain an overview of the risks and benefits of oversizing and to determine the optimal degree of oversizing of stent grafts used for endovascular abdominal aortic aneurysm repair. METHODS: PUBMED, EMBASE and Cochrane Library databases were searched for articles related to the impact of proximal endograft oversizing on complications after endovascular aneurysm repair. After in- and exclusion, 23 relevant articles reporting on 8415 patients remained for analysis and critical appraisal. RESULTS: Most studies that investigated neck dilatation are flawed by poor methodology. No clear relationship between proximal oversizing and neck dilatation relative to the first post-operative scan was found. None of the studies described a positive relationship between the degree of oversizing and the incidence of endoleaks. On the contrary, oversizing up to 25% seems to decrease the risk of proximal endoleaks. There are conflicting data regarding the risk of graft migration when oversizing by more than 30%. CONCLUSIONS: Based on the best available evidence, the current standard of 10-20% oversizing regime appears to be relatively safe and preferable. Oversizing >30% might negatively impact the outcome after EVAR. Studies of higher quality are needed to further assess the relationship between proximal oversizing and the incidence of complications, particularly regarding the impact on aneurysm neck dilatation.

Aortic pulsatile distention in young healthy volunteers is asymmetric: analysis with ECG-gated MRI.

OBJECTIVE: Knowledge of aortic shape changes throughout the cardiac cycle can offer improved understanding of vascular pathophysiology and may have crucial impact on stentgraft design and EVAR durability. To understand underlying mechanisms of dynamic changes in aortic aneurysm (neck) morphology, the undiseased aorta has to be studied first. Objective is to visualize and characterize dynamic aortic shape changes in young healthy volunteers. MATERIALS AND METHODS: Fifteen healthy volunteers (7 male, median age 24 year, range 18-28) were scanned using ECG-gated balanced gradient-echo MRI, with 16 reconstructed cardiac phases. Transverse scans were made perpendicular to the aorta: (A) above the aortic bifurcation, (B) infrarenal, (C) juxtarenal, (D) suprarenal and (E) above the celiac trunk. After aortic lumen segmentation, radial changes during the cardiac cycle were measured, from the center of mass, over 360 degrees, and plotted. An ellipse was fitted over the distention plots, yielding the direction (AP:0 degrees, Right: -90 degrees, Left: 90 degrees ) and magnitude of radius change over the major and minor axis. RESULTS: Asymmetric distention was observed, with a variable rate per patient and level. Radius changes decreased from the proximal to distal aorta. Radius changes over the major axis ranged from 14% to 41%. At level A mean change in radius over the minor versus major axis was 1.4+/-0.2mm (17%) versus 1.6+/-0.2mm (20%), respectively. At B 1.7+/-0.4mm (22%) versus 2.0+/-0.4mm (25%), at C 1.7+/-0.4mm (22%) versus 2.2+/-0.4mm (27%) at D 2.0+/-0.4mm (25%) versus 2.4+/-0.5mm (30%) and at E 2.2+/-0.3mm (27%) versus 2.6+/-0.3mm (32%). Mean orientation of the major axis was (A) 0.8+/-23.3 degrees , (B) 1.8+/-31.3 degrees , (C) 14.0+/-15.5 degrees , (D) -28.8+/-48.0 degrees and (E) 18.4+/-22.2 degrees. CONCLUSIONS: Aortic pulsatile distention in young healthy volunteers is asymmetric, with up to 41% radius change in the descending aorta. This study offers a frame of reference for dynamic imaging studies in patients with aortic pathology and provides a valuable non-invasive tool for future research into aortic distention, development and localization of vascular pathology.

Multidimensional characterization of carotid artery stenosis using CT imaging: a comparison with ultrasound grading and peak flow measurement.

PURPOSE: Clinical decision making for carotid surgery depends largely upon stenosis grade. While digital subtraction angiography remains the gold standard for stenosis grading, many physicians use less invasive modalities. The purpose of this study was to compare the results of multidimensional Computed tomography (CTA) with ultrasound (US) grading and peak flow velocity (PSV). METHODS: 37 stenosed carotid arteries were studied retrospectively in 36 consecutive patients. US grading and PSV were compared to multidimensional CTA analysis (diameter, area and volumetric measurements), performed by a medical software company. Calculations of stenosis percentage on CTA were made using the NASCET and ECST methodology. Diameter measurements were also performed by a neuroradiologist. RESULTS: All CTA diameter, area and volume measurements had only modest correlation with PSV (r<0.5) and ultrasound grading (p<0.5). There was concordant classification of stenosis grades in only 40-60% of cases. CTA diameter, area and volume measurements had good correlation (0.69

Dynamic cine-CT angiography for the evaluation of the thoracic aorta; insight in dynamic changes with implications for thoracic endograft treatment.

OBJECTIVE: Thoracic aneurysm preoperative imaging is performed using static techniques without consideration of normal aortic dynamics. Improved understanding of the native aortic environment into which thoracic endografts are placed may aid in device selection. It is unclear what comprises normal thoracic aortic pulsatility. We studied these phenomena dynamically using ECG-gated 64-slice CTA. METHODS: Maximum diameter and area change per cardiac cycle was measured at surgically relevant anatomic thoracic landmarks in ten patients; 1.0 cm proximal and distal to the subclavian artery, 3.0 cm distal to the subclavian artery, and 3.0 cm proximal to the celiac trunk. Data was acquired using a novel ECG-gated dynamic 64-slice CT scanner during a single breath hold with a standard radiation dose and contrast load. Eight gated data sets, covering the cardiac cycle were reconstructed, perpendicular to the central lumen. RESULTS: There is impressive change in both maximum diameter and area in the thoracic aorta during the cardiac cycle. Mean maximum diameter changes of greater than 10% are observed in the typical sealing zones of commercially available endografts corresponding to diameter increases of up to 5mm. Aortic area increases by over 5% per cardiac cycle. CONCLUSIONS: ECG-gated dynamic CTA with standard radiation dose is feasible on a 64-slice scanner and provides insight into (patho) physiology of thoracic aortic conformational changes. Clinicians typically oversize thoracic endografts by 10%. With aortic pulsatility resulting in diameter changes of up to 17.8%, the potential exists for endograft undersizing, graft migration, intermittent type I endoleak, and poor patient outcome. Furthermore, aortic pulsatility is not evenly distributed, and non-circular stentgraft designs should be considered in the future since aortic distension in the aneurysm neck is not evenly distributed.