Development and Validation of a Novel Anaerobic Carbapenem Inactivation Method (Ana-CIM) for the Detection of Carbapenemase Production in Bacteroides fragilis.

Antibiotic resistance, particularly to carbapenems, is of increasing concern in Bacteroides fragilis. Carbapenem resistance in B. fragilis is most often mediated by the activation of chromosomally encoded metallo-ß-lactamase cfiA by the presence of an upstream insertion sequence (IS). While traditional phenotypic susceptibility methods and molecular tests to detect carbapenem resistance in B. fragilis exist, they are not available in most clinical microbiology laboratory settings. Here, we describe the development of the anaerobic carbapenem inactivation method (Ana-CIM) for predicting carbapenemase production in B. fragilis based off the principles of the well-established modified carbapenem inactivation method (mCIM) for Enterobacterales and Pseudomonas aeruginosa. We also present the clinical validation and reproducibility of the Ana-CIM at three clinical laboratory sites (with 60 clinical isolates, 45% ertapenem resistant). Compared to ertapenem susceptibility by Etest interpreted by CLSI M100 Ed30, the Ana-CIM accurately detected carbapenem resistance in B. fragilis with categorical agreement (CA) of 87% (52/60) and 0% (0/21) very major error (VME), 11% (4/36) major error (ME), and 7% (4/60) minor error (mE) rates across all sites. Additionally, the Ana-CIM demonstrated high reproducibility with 5 clinical and 3 quality control (QC) isolates tested in triplicate with 3 commercial Mueller-Hinton media across all sites, with 93% (604/648) of replicates within a 2-mm zone size of the mode for each isolate. We conclude that the Ana-CIM can be readily deployed in clinical laboratories at a low cost for detection of carbapenemase-mediated resistance in B. fragilis.

Clostridioides difficile Whole-genome Sequencing Differentiates Relapse With the Same Strain From Reinfection With a New Strain.

BACKGROUND: Current approaches in tracking Clostridioides difficile infection (CDI) and individualizing patient management are incompletely defined. METHODS: We recruited 468 subjects with CDI at Mayo Clinic Rochester between May and December 2016 and performed whole-genome sequencing (WGS) on C. difficile isolates from 397. WGS was also performed on isolates from a subset of the subjects at the time of a recurrence of infection. The sequence data were analyzed by determining core genome multilocus sequence type (cgMLST), with isolates grouped by allelic differences and the predicted ribotype. RESULTS: There were no correlations between C. difficile isolates based either on cgMLST or ribotype groupings and CDI outcome. An epidemiologic assessment of hospitalized subjects harboring C. difficile isolates with ≤2 allelic differences, based on standard infection prevention and control assessment, revealed no evidence of person-to-person transmission. Interestingly, community-acquired CDI subjects in 40% of groups with ≤2 allelic differences resided within the same zip code. Among 18 subjects clinically classified as having recurrent CDI, WGS revealed 14 with initial and subsequent isolates differing by ≤2 allelic differences, suggesting a relapse of infection with the same initial strain, and 4 with isolates differing by >50 allelic differences, suggesting reinfection. Among the 5 subjects classified as having a reinfection based on the timing of recurrence, 3 had isolates with ≤2 allelic differences between them, suggesting a relapse, and 2 had isolates differing by >50 allelic differences, suggesting reinfection. CONCLUSIONS: Our findings point to potential transmission of C. difficile in the community. WGS better differentiates relapse from reinfection than do definitions based on the timing of recurrence.

Clinical utility of anaerobic culture of cerebrospinal fluid.

Anaerobic meningitis is a rare serious clinical condition which mainly affects vulnerable populations and patients with predisposing factors such as head trauma, prior neurosurgical procedures or implantable medical devices such as ventriculoperitoneal shunts or ventricular drains. In this study we retrieved data from aerobic and anaerobic cultures of cerebrospinal (CSF) or ventricular fluid ordered over a 5 year period at our institution. A total of 8868 aerobic and 594 anaerobic cultures were performed from 2013 to 2017. 24/594 (4%) anaerobic cultures from 14 patients were positive for anaerobes. Only 3 of those patients were diagnosed clinically with anaerobic meningitis, each with predisposing factors, while anaerobes (Cutibacterium acnes and Clostridium perfringens) recovered from the remaining 21 patients were regarded as contaminants. 129/8868 (1.45%) aerobic CSF cultures were positive for anaerobes. 120/129 (93%) cultures recovered C. acnes while non-C. acnes anaerobes were recovered in the remaining 9 cultures and were deemed to be contaminants. In the majority of situations, recovery of C. acnes from CSF or ventricular fluid was regarded as contamination. Our cohort included 18 patients with a ventriculoperitoneal shunt or ventricular drain, 17 of whom had C. acnes recovered from either aerobic or anaerobic culture, and 10 were treated with targeted antibiotics and surgical replacement of the shunt or drain. Anaerobic culture of the CSF or ventricular fluid aided in identification of two patients with anaerobic meningitis and an additional two patients with shunt infection. Anaerobe culture of CSF is important in identification of anaerobic meningitis, as growth of anaerobes other than C. acnes is rare from aerobic CSF culture.

Routine use of MALDI-TOF MS for anaerobic bacterial identification in clinical microbiology.

In 2013, we adopted MALDI-TOF MS using the Bruker Biotyper system for identification of anaerobic bacteria into our routine clinical practice. Here, we describe our experience with the use of MALDI-TOF MS for anaerobic bacterial identification, highlighting its value in replacing the more costly and time-consuming 16S ribosomal RNA gene PCR plus sequencing-based approach as the primary method of anaerobic bacterial identification. We also describe our more recent experience with the use of early/rapid MALDI-TOF MS for identification of anaerobic bacteria performed on short incubation (4-6 h) plated aerobic media from anaerobic blood culture bottles positive for Gram-negative bacilli.

Laboratory Workflow Analysis of Culture of Periprosthetic Tissues in Blood Culture Bottles.

Culture of periprosthetic tissue specimens in blood culture bottles is more sensitive than conventional techniques, but the impact on laboratory workflow has yet to be addressed. Herein, we examined the impact of culture of periprosthetic tissues in blood culture bottles on laboratory workflow and cost. The workflow was process mapped, decision tree models were constructed using probabilities of positive and negative cultures drawn from our published study (T. N. Peel, B. L. Dylla, J. G. Hughes, D. T. Lynch, K. E. Greenwood-Quaintance, A. C. Cheng, J. N. Mandrekar, and R. Patel, mBio 7:e01776-15, 2016, https://doi.org/10.1128/mBio.01776-15), and the processing times and resource costs from the laboratory staff time viewpoint were used to compare periprosthetic tissues culture processes using conventional techniques with culture in blood culture bottles. Sensitivity analysis was performed using various rates of positive cultures. Annualized labor savings were estimated based on salary costs from the U.S. Labor Bureau for Laboratory staff. The model demonstrated a 60.1% reduction in mean total staff time with the adoption of tissue inoculation into blood culture bottles compared to conventional techniques (mean ± standard deviation, 30.7 ± 27.6 versus 77.0 ± 35.3 h per month, respectively; P < 0.001). The estimated annualized labor cost savings of culture using blood culture bottles was $10,876.83 (±$337.16). Sensitivity analysis was performed using various rates of culture positivity (5 to 50%). Culture in blood culture bottles was cost-effective, based on the estimated labor cost savings of $2,132.71 for each percent increase in test accuracy. In conclusion, culture of periprosthetic tissue in blood culture bottles is not only more accurate than but is also cost-saving compared to conventional culture methods.

Blame it on patriarchy: more sexist attitudes are associated with stronger consideration of cosmetic surgery for oneself and one's partner.

In the present work, we examined associations between oppressive, sexist beliefs and consideration of cosmetic surgery for oneself and also endorsement of cosmetic surgery for one's romantic partner. A total of 554 German-speaking volunteers from the community, mainly in Austria, completed measures of consideration of cosmetic surgery and three measures of sexist attitudes, while a subset of participants in romantic relationships completed a measure of endorsement of cosmetic surgery for their partners along with the measures of sexism. Preliminary analyses showed that women and single respondents were more likely to consider having cosmetic surgery than men and committed respondents, respectively. Further analyses showed that consideration of cosmetic surgery for oneself was significantly associated with sexist attitudes, particularly hostile attitudes to women. In addition, among participants in a relationship, sexist attitudes were associated with endorsement of cosmetic surgery for one's partner. These results indicate that attitudes to cosmetic surgery for oneself and one's partner are shaped by gender-ideological belief systems in patriarchal societies. Possible implications for understanding the motivations for having cosmetic surgery, among both single respondents and couples, are discussed.