Search

Background: Environmental sampling and detection methods for fungi in healthcare settings are not well-established. We previously refined methods for fungal sampling and detection in a controlled laboratory environment and aimed to validate them in a real-world healthcare setting. Methods: We performed a microbiological analysis of air and surfaces in three inpatient units at a tertiary care center. Surface samples were obtained with foam sponges from 3 locations in patient rooms (Patient bedrails, bathroom floor, HVAC export) and 5 locations in units (HVAC exports 3x, clean linen storage, soiled linen storage). Air samples were taken with an active air sampler directly below HVAC exports. Sponges were processed using the stomacher technique. Samples underwent DNA extraction followed by qPCR with FungiQuant primers targeting the 18S rRNA gene. Amplicons from positive samples were sequenced (NextSeq 1000, 300bp PE) and SmartGene databases were used to interpret sequence data. For comparison to culture methods, samples were also plated onto Sabouraud and HardyCHROM Candida + auris medias. Fungal growth underwent DNA extraction, 18S PCR and Sanger sequencing for genus and species identification. Results: A total of 85 samples were obtained, from 15 patient rooms and three units resulting in 61 surface and 24 air samples. Patients in study rooms had a median age of 53, 9 (60%) were male, and no patients had an invasive fungal infection during their hospital encounter. 44 (53%) and 39 (46%) samples were positive for fungi via qPCR and culture, respectively. Of the 44 positive qPCR samples, microbiome analyses identified at least one fungi to the species, genus and family levels in 43 (98%), 28 (64%), 18 (41%) samples, respectively (Table 1). 114 total isolates were identified of which the most common were Mallassezia restricta (30 [26%]), Malassezia globose (29 [25%]), and Pennicillium paradoxum (4 [4%]). 39 genera were identified of which the most common were Mucor (19 [49%]) and Candida (8 [21%]). Of the 39 culture positive samples, 90 total isolates were recovered. The most common species were Paradendryphiella arenariae (19 [21%]), Aspergillus niger (12 [13%]) and Penicillium commune (12 [13%]). Conclusion: These results demonstrate the presence of diverse fungal species in both air and surface samples across inpatient units. Higher sensitivity was noted utilizing qPCR, however, identified genera and species were markedly different between qPCR and culture methods. Larger studies are needed to assess the efficacy of qPCR for fungal detection in the healthcare environment.

Background: Frequently used physical therapy (PT) equipment is notably difficult to disinfect due to equipment material and shape, however, the efficacy of standard disinfection of PT equipment is poorly understood. Methods: We completed a prospective observational microbiological analysis of fomites used in adult or pediatric PT at Duke University Health System, Durham, North Carolina, from September to December 2022. Predetermined study fomites were obtained after being used during a clinical shift and standard disinfection had been completed by clinical service staff. Fomites were split into 2 halves, left and right, for sampling. Samples were taken with premoistened cellulose sponges processed using the stomacher technique and were incubated on appropriate selective and general medias. We defined antimicrobial-resistant, clinically important pathogens (AMR-CIP) as MRSA, VRE, and MDR-gram-negative isolates, and non–AMR-CIP as MSSA, VSE, and gram-negative species. Study fomites were grouped as follows: (1) pediatric pig toy, (2) walking aids (walkers or canes), (3) balls (medicine, dodge, etc), and (4) other (foam roller, sliding board, etc). Results: In total, 47 patients, 61 fomites, and 122 were analyzed. Of the study patients, 24 (51%) were female, 13 (27%) had active infections, and 15 (32%) were on contact precautions. Because fomites were split in half, patients in the left and right study arms were identical. Overall, the median total colony-forming-units (CFU) of study fomites was 1,348 (IQR, 398–2,365): 468 (IQR, 161–1,230) for the left side study arm and 540 (IQR, 102–1,221) for the right study arm (P = .45). At the sample level, 52 (43%), 15 (12%), and 37 (30%) of 122 samples harbored any CIPs, AMR CIPs, or non-AMR CIPs, respectively. At the fomite level, 27 (44%), 5 (8%), 15(25%), and 7 (11%) of 61 fomites harbored any CIPs, only AMR-CIPs, only non-AMR CIPs, or both AMR and non-AMR CIPs, respectively. Generally, therapy balls were the most contaminated study fomites (n = 2,237; IQR, 1,425–2,658), and walking aids were most frequently contaminated with any CIPs (n = 26, 72%), AMR CIPs (n = 8, 22%), and non-AMR CIPs (n = 15, 47%). Discussion: Following routine disinfection, frequently used PT equipment remained heavily contaminated and harbored AMR and non-AMR CIPs, supporting the notion that PT equipment is difficult to disinfect via standard disinfection. Additionally, left-, and right-side fomite divisions had similar pathogens, suggesting that this sampling model of intrapatient comparisons may be helpful for resolving case-mix issues in future studies. Future work should focus on PT-specific enhanced disinfection strategies to improve the disinfection of PT equipment.

Background: SARS-CoV-2 N95 mask contamination in healthcare providers (HCPs) treating patients with COVID-19 is poorly understood. Method: We performed a prospective observational study of HCP N95 respirator SARS-CoV-2 contamination during aerosol-generating procedures (AGPs) on SARS-CoV-2–positive patients housed in a COVID-19–specific unit at an academic medical center. Medical masks were used as surrogates for N95 respirators to avoid waste and were worn on top of HCP N95 respirators during study AGPs. Study masks were provided to HCPs while donning PPE and were retrieved during doffing. Additionally, during doffing, face shields were swabbed with Floq swabs premoistened with viral transport media (VTM) prior to disinfection. Medical masks were cut into 9 position-based pieces, placed in VTM, vortexed, and centrifuged (Fig. 1). RNA extraction and RT-PCR were completed on all samples. RT-PCR–positive samples underwent cell culture infection to detect cytopathic effects (CPE). Contamination was characterized by mask location and front and back of face shields. Patient COVID-19 symptoms were collected from routine clinical documentation. Study HCPs completed HCP-role–specific routine care (eg, assessing, administering medications, and maintaining oxygen supplementation) while in patient rooms and were observed by study team members. Results: We enrolled 31 HCPs between September and December 2021. HCP and patient characteristics are presented in Table 1. In total, 330 individual samples were obtained from 31 masks and 26 face shields among 12 patient rooms. Of the 330 samples, 0 samples were positive for SARS-CoV-2 via RT-PCR. Positive controls were successfully performed in the laboratory setting to confirm that the virus was recoverable using these methods. Notably, all samples were collected from HCPs caring for COVID-19 patients on high-flow, high-humidity Optiflow (AGP), with an average of 960 seconds (IQR, 525–1,680) spent in each room. In addition to Optiflow and routine care, study speech pathologists completed an additional AGP of fiberoptic endoscopic evaluation of swallowing. Notably, 29 (94%) of 31 study HCP had physical contact with their patient. Conclusions: Overall, mask contamination in HCPs treating patients with COVID-19 undergoing AGPs was not detectable while wearing face shields, despite patient contact and performing AGP.

Background: Enhanced strategies for daily disinfection in acute-care hospital rooms are needed but are poorly understood. Methods: We conducted a randomized control trial pilot study in acute-care hospital rooms at Duke University Health System in Durham, North Carolina, comparing the efficacy of a novel EPA-registered quaternary ammonium disinfectant with 24-hour activity, Sani24, to routine daily disinfection. Rooms housing patients on contact precautions were enrolled. In each study room, the bedrails, overbed table, and sink were divided into 2 equal halves, or sides, labeled left and right, with sample areas of 2,000 cm2, 1,750 cm2, and 400 cm2, respectively. Each sample area side was then randomized 1:1 to intervention or control by a coin toss. Sani24 was applied to the surface of each intervention sample side and allowed to air dry. Control sides were left alone. Environmental services (EVS) staff were not involved in the study and were blinded to randomization status. Glogerm dots were applied to all 6 sample-area sides after application of the intervention to measure compliance of daily disinfection by EVS and the removal of the intervention agent. Microbiological samples were taken with sponges premoistened with neutralizing buffer from each sample area side for 6 total samples (3 intervention and 3 control) immediately before and 24 hours following application of the intervention agent. Clinically important pathogens (CIP) were defined as MRSA, VRE, and CRE. The primary outcome was room CFU on study day 1, which was compared using a Wilcoxon rank-sum test. Results: In total, 20 patient rooms were enrolled in the study, and 240 samples were obtained from 120 sites (60 intervention and 60 control) from November 2021 to January 2022. Enrolled patients were all on contact isolation and had an active infection; 15 (75%) were bedridden and 8 (40%) were female. On day 0, baseline contamination was similar between study arms: 7,460 (IQR,4,204–16,482) room CFU and 18 samples (30%) harboring CIP in the intervention arm versus 7,273 (IQR, 3,142–21,117) and 15 samples (25%) in the control arm (P = .49 and .47, respectively). On day 1, intervention areas had significantly lower CFU at 4,016 (IQR, 2,339–7,358) compared to controls at 6,112 CFU (IQR, 3,484–11,356; P = .01). No significant differences were detected between study arms regarding CIP recovery. Glogerm was minimally removed from sample areas (n = 7, 3%), and the result was similar between study arms. Conclusions: The use of the quaternary ammonium disinfectant with 24-hour activity on high-touch healthcare surfaces led to reduced contamination over a 24-hour period. Routine daily disinfection compliance by EVS was low since minimal sample areas had Glogerm removed