Impact of a real-time diagnostic and antimicrobial stewardship workflow on time to appropriate therapy for infections caused by multidrug-resistant Gram-negative organisms.

INTRODUCTION: Multidrug-resistant (MDR) Gram-negative organisms cause life-threatening infections, and the incidence is rising globally. Timely therapy for these infections has a direct impact on patient survival. This study aimed to determine the impact of a multidisciplinary diagnostic and antimicrobial stewardship (AMS) workflow on time to appropriate therapy (TAP) for these infections using novel beta-lactam/beta-lactamase inhibitors. METHODS: This was a retrospective quasi-experimental study of adult patients with carbapenem-resistant Enterobacterales (CRE) and multidrug-resistant Pseudomonas (MDR PsA) infections at a 1500 bed university hospital. Included patients who received ≥ 72 hours of ceftazidime-avibactam (CZA) or ceftolozane-tazobactam (C/T) from December 2017 to December 2019. During the pre-intervention period (December 2017 to December 2018), additional susceptibilities (including CZA and C/T) were performed only upon providers' request. In 2019, reflex algorithms were implemented for faster identification and testing of all CRE/MDR PsA isolates. Results were communicated in real-time to the AMS team to tailor therapy. RESULTS: A total of 99 patients were included, with no between-group differences at baseline. The median age was 60 years and 56 (56.7%) were in intensive care at the time of culture collection. Identified organisms included 71 (71.7%) MDR PsA and 26 CRE, of which 18 were carbapenemase producers (Klebsiella-producing carbapenemase = 12, New Delhi metallo-ß-lactamase = 4, Verona integron-encoded metallo-ß-lactamase = 2). The most common infections were pneumonia (49.5%) and bacteraemia (30.3%). A decrease was found in median TAP (103 [IQR 76.0-156.0] vs. 75 [IQR 56-100] hours; P < 0.001). Median time from culture collection to final susceptibility results was shorter in the post-intervention group (123 vs. 93 hours; P < 0.001). CONCLUSION: This study identified improvement in TAP in MDR PsA and CRE infections with implementation of a reflex microbiology workflow and multidisciplinary antimicrobial stewardship initiatives.

Mycobacterium abscessus Infections in Solid Organ Transplant Recipients: Single-Center Experience in the United States, 2013-2018.

Background: Mycobacterium abscessus is increasingly recognized as a human pathogen causing life-threatening infections in immunocompromised patients. There is a paucity of data around this topic in solid organ transplant (SOT) recipients. Methods: This work was a single-center retrospective cohort study of all SOT recipients with a positive culture for M abscessus between 2013 and 2018. Results: A total of 20 patients (55% female) met inclusion criteria, including 1 kidney recipient (5.0%), 2 liver recipients (10.0%), 12 lung recipients (60.0%), 1 heart recipient (5.0%), and 4 combined organ recipients (20.0%). The median time from SOT to infection was 100 days (range, 30-431 days). Thirteen (65.0%) patients (1 kidney, 1 heart, 7 lung, 1 liver, 1 intestine, and 2 multivisceral) were treated with a median duration of 185 antibiotic days (range, 20-523 days). Among them, M abscessus was isolated from respiratory samples in 8 and nonrespiratory samples in 5; 4 of 13 (30.8%) patients had treatment failure and 3 of 13 (23.1%) had unrelated deaths within 1 year after diagnosis. Seven patients (5 lung transplant recipients) with the organism isolated from respiratory samples were not treated as their cultures represented airway colonization or contamination; of those, 2 (28.6%) died (unrelated to infection) and 5 (71.4%) were alive without the infection after 1 year of follow-up. Conclusions: Mycobacterium abscessus infections affect SOT recipients with a high proportion of clinical failures. However, in lung recipients, not all positive cultures correlated with infection, and without treatment some patients had good clinical outcomes. Thus, differentiating colonization from infection is important, and infection prevention measures and novel therapeutic agents are needed for SOT recipients.

Implementing carbapenem-resistance testing algorithms for Enterobacteriales and Pseudomonas aeruginosa: diagnostic and antimicrobial stewardship with timely infection prevention.

Early detection of carbapenem resistance in Gram-negative (CRGN) infections can help tailor therapy sooner and prevent horizontal transmission more efficiently. This involves timely, multidisciplinary communication. To this aim, we describe our experience with implementing reflex testing algorithms for expanded antibiotic susceptibilities and molecular mechanisms of resistance in a large academic hospital.

Successful Treatment of Invasive Fungal Infection Due to Highly Resistant Aspergillus calidoustus in an Allogeneic Hematopoietic Cell Transplant Recipient.

Invasive aspergillosis (IA) is the most common invasive fungal infection following a hematopoietic cell transplant, with emerging cryptic species exhibiting resistance to commonly used antifungals such as azoles. These species have been increasingly found after the introduction of anti-mold prophylaxis. We report a case of a 56-year-old female with primary myelofibrosis whose allogeneic hematopoietic cell transplant was complicated by disseminated fungal infection (skin, lung) due to Aspergillus calidoustus, a cryptic specie. Treatment of Aspergillus species remains challenging as these cryptic species are usually resistant to azoles including voriconazole which is the first line of treatment of IA. Infection was successfully treated with surgical excision and combination antifungal therapy based on in vitro susceptibility and synergy testing. Therapy included isavuconazole, a drug that has been shown to be non-inferior to voriconazole in the treatment of invasive mold infections.

Next-generation sequencing of microbial cell-free DNA for rapid noninvasive diagnosis of infectious diseases in immunocompromised hosts.

Background: Cell-free DNA (cfDNA) sequencing has emerged as an effective laboratory method for rapid and noninvasive diagnosis in prenatal screening testing, organ transplant rejection screening, and oncology liquid biopsies but clinical experience for use of this technology in diagnostic evaluation of infections in immunocompromised hosts is limited.  Methods: We conducted an exploratory study using next-generation sequencing (NGS) for detection of microbial cfDNA in a cohort of ten immunocompromised patients with febrile neutropenia, pneumonia or intra-abdominal infection.  Results: Pathogen identification by cfDNA NGS demonstrated positive agreement with conventional diagnostic laboratory methods in 7 (70%) cases, including patients with proven/probable invasive aspergillosis, Pneumocystis jirovecii pneumonia, Stenotrophomonas maltophilia bacteremia, Cytomegalovirus and Adenovirus viremia. NGS results were discordant in 3 (30%) cases including two patients with culture negative sepsis who had undergone hematopoietic stem cell transplant in whom cfDNA testing identified the potential etiological agent of sepsis; and one kidney transplant recipient with invasive aspergillosis who had received >6 months of antifungal therapy prior to NGS testing. Conclusion: These observations support the clinical utility of measurement of microbial cfDNA sequencing from peripheral blood for rapid noninvasive diagnosis of infections in immunocompromised hosts. Larger studies are needed.

Carbapenemase-Producing Raoultella Planticola: A Rare Cause of Pneumonia and Bacteremia.

Raoultella planticola is a gram-negative bacterium of the Enterobacteriaceae family that is usually found in soil, plant and aquatic environments. It is an uncommon human pathogen and has been associated with cases of bacteremia, pneumonia, urinary tract infections, among others. Here, we present the case of an 85-year-old female that developed nosocomial pneumonia and bacteremia caused by Raoultella planticola. Pertinent microbiological studies detected carbapenemase production codified by the blaKPC gene. The patient was successfully treated with ceftazidime/avibactam and polymyxin. Our case illustrates the pathogenic potential of this organism and highlights the importance of phenotypic and genotypic assays for the appropriate identification of carbapenemase production.

Invasive Rhinosinusitis Caused by Lasiodiplodia theobromae in an Allogeneic Hematopoietic Cell Transplant Recipient Case Report and Review of Literature.

Lasiodiplodia theobromae is a known plant pathogen in tropical and subtropical areas. Few cases have been reported in humans (usually keratitis and endophthalmitis) with only two cases of fungal sinusitis in immunocompromised and immunocompetent patients published to date. We report a case of invasive sinusitis secondary to L. theobromae in an allogeneic hematopoietic cell transplant recipient successfully treated with surgical debridement and triazole antifungals with a review of available literature.

A comparison of two microbial detection methods used in aseptic processing of musculoskeletal allograft tissues.

Tissues from 78 musculoskeletal donors were concurrently tested for microorganisms using both a swab and liquid culture method. An aggregate total of 20 organisms were detected by both methods. The swab detected 4/20 organisms while the liquid culture detected 18/20 organisms. The swab method yielded sensitivity and negative predictive values of 20 and 92.3%, respectively. Comparatively, the liquid culture displayed a sensitivity of 90% and a negative predictive value of 99%. These results clearly demonstrate that the liquid culture method is superior to swab cultures in microbial detection. Additional studies are necessary to determine the optimal culture conditions for different types of tissues when utilizing the liquid culture method.

Detection of group B streptococci in Lim broth by use of group B streptococcus peptide nucleic acid fluorescent in situ hybridization and selective and nonselective agars.

The sensitivity, specificity, and positive and negative predictive values for the detection of group B streptococci from Lim enrichment broth with sheep blood agar (SBA), with selective Streptococcus agar (SSA), and by a peptide nucleic acid fluorescent in situ hybridization (PNA FISH) assay were as follows: for culture on SBA, 68.4%, 100%, 100%, and 87.9%, respectively; for culture on SSA, 85.5%, 100%, 100%, and 94.1%, respectively; and for the PNA FISH assay, 97.4%, 98.3%, 96.1%, and 98.9%, respectively.

Efficacy of linezolid versus vancomycin in the treatment of methicillin-resistant Staphylococcus aureus discitis: a controlled animal model.

STUDY DESIGN: A rabbit model was used to assess the efficacy of linezolid and vancomycin for the treatment of discitis due to methicillin-resistant Staphylococcus aureus (MRSA). Nontreated controls were used for comparison. OBJECTIVE: The purpose of this study was to determine if there was a therapeutic difference between using linezolid and vancomycin in the treatment of MRSA discitis. SUMMARY OF BACKGROUND DATA: Vancomycin is currently the gold standard treatment for medical management of MRSA discitis. Linezolid is a relatively new drug that has been approved for treatment of MRSA infections, but currently there is no research demonstrating its efficacy at treating infections of the disc space. METHODS: Twenty-four rabbits were inoculated with MRSA at two adjacent lumbar disc spaces via an anterior retroperitoneal approach. Six rabbits were to receive only pain medication and to serve as controls. Ten rabbits were assigned to a 5-day course of intravenous vancomycin, and 8 were assigned to a 5-day course of intravenous linezolid. Disc spaces were sent for quantitative culture after the 5-day treatment course. RESULTS: The mean culture growth for the disc spaces was not statistically different between the linezolid treated group and the nontreated controls. While vancomycin treatment did lead to lower bacterial loads when compared with controls, the reduction was not statistically significant. When bacterial counts for the vancomycin group and linezolid group were compared, vancomycin treatment resulted in less bacterial growth. This difference was statistically significant. CONCLUSIONS: Linezolid is a clinically attractive alternative to vancomycin due to its mild side effect profile and oral bioavailability. However, in this MRSA discitis model with a short treatment course, vancomycin was superior to linezolid.

Variations in lethal toxin and cholesterol-dependent cytolysin production correspond to differences in cytotoxicity among strains of Clostridium sordellii.

Clostridium sordellii is an emerging human pathogen and frequent contaminant of cadaver-derived tissue transplant material. Herein, we provide data suggesting the potential for severe C. sordellii-associated disease may be dictated by whether the specific strain produces lethal toxin (TcsL) or sordellilysin (SDL), a cholesterol-dependent cytolysin. The virulence factor profiles of 14 C. sordellii isolates were determined, and culture supernatant from six of the isolates was found to be cytotoxic to mammalian cells; yet, only one of these strains conferred cytotoxicity via production of TcsL. Cytotoxicity of TcsL- strains correlated with the production of sordellilysin, which was also recognized by an antiperfringolysin O antibody. However, supernatant from TcsL+, SDL- strains demonstrated a lower LD50 relative to TcsL-, SDL+ strains, suggesting the potential for severe C. sordellii-associated disease may be determined by the particular strain colonizing the host.

Antifungal penetration into normal rabbit nucleus pulposus.

STUDY DESIGN: A rabbit model was used to assess the penetration into the nucleus pulposus of 3 commonly used antifungal medications: amphotericin B, amphotericin B lipid complex, and fluconazole. OBJECTIVES: The purpose of this study was to quantitate the penetration of antifungal medications into the normal rabbit nucleus pulposus. SUMMARY OF BACKGROUND DATA: Fungal infections of the spine are rarely, if ever, treated with medical management alone. Although antibiotic penetration into the nucleus pulposus has been studied extensively, no previous studies have attempted to quantitate the penetration of antifungals into the nucleus pulposus. METHODS: Twenty-four rabbits were given 2 doses of 1 of the antifungal medications studied. One hour after completion of the second dose, the animal was killed and the thoracolumbar spine was excised en bloc. Specimens of nucleus pulposus and serum were obtained and sent to an outside laboratory for analysis. Gas chromatography was used to determine the fluconazole tissue levels, and a bioassay was used to measure amphotericin B tissue levels. RESULTS: Three animals in the amphotericin B group died either after the first or second dose of medication was administered. Although amphotericin B and amphotericin B lipid complex did not show adequate penetration into the nucleus pulposus in 12 out of 12 animals, fluconazole reached therapeutic tissue levels in 5 out of 7 animals. CONCLUSIONS: Fluconazole showed superior penetration into the nucleus pulposus in an uninfected rabbit model when compared to amphotericin B and amphotericin B lipid complex. These findings were found to be statistically significant (P = 0.021), and they suggest that fluconazole may be a better choice for empiric therapy of fungal spine infections while cultures and sensitivities are pending.

Blood and marrow cultures as indicators of bone contamination in cadaver donors.

Of 770 cadaver bone donors evaluated, 185 had positive blood or ilium marrow aspirate cultures. These donors were matched with an immediately preceding or subsequent donor with negative blood and marrow cultures. Donors with cultures positive for skin contaminants only were not included in the study. Samples of the blood and bone marrow, surface swab cultures, and cultures of tissue samples of the excised skeletal tissues were obtained at the time of tissue procurement. There were 88 (48%) donors with similar microbial species recovered from the blood and ilium marrow. These donors had a higher rate of positive bone cultures (30%) than donors with positive blood (15%) or marrow cultures only (11%) or donors with negative blood and marrow culture results (7.3%). Recovery of similar isolates from the blood and marrow had a positive predictive value of 72% for the isolation of the same types of organisms from the excised tissues compared with 38% for donors with positive blood cultures only. Although not absolute predictors of tissue contamination, combined blood and bone marrow cultures were more reliable indicators of tissue contamination than blood cultures alone.