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Negative Pressure Wound Therapy (NPWT) has been widely adopted in wound healing strategies due to its multimodal mechanism of action. While NPWT’s positive impression on wound healing is well-established, its effect on bacterial load reduction remains equivocal. This study investigates NPWT’s efficacy in reducing bioburden using an in vitro porcine skin model, focusing on the impact of Staphylococcus aureus and Staphylococcus epidermidis. Custom-made negative pressure chambers were employed to apply varying negative pressures. Porcine skin was cut into 5 × 5 cm squares and three standardized wounds of 6 mm each were created using a biopsy punch. Then, wounds were infected with S. aureus and S. epidermidis bacterial suspensions diluted 1:10,000 to obtain a final concentration of 1.5 × 104 CFU/ml and were placed in negative pressure chambers. After incubation, bacterial counts were expressed as colony-forming units (CFU) per ml. For S. aureus at 120 hours, the median CFU, mean area per colony, and total growth area were notably lower at −80 mmHg when compared to −250 mmHg and −50 mmHg, suggesting an optimal negative pressure for the pressure-dependent inhibition of the bacterial proliferation. While analyzing S. epidermidis at 120 hours, the response to the negative pressure was similar but less clear, with the minor CFU at −100 mmHg. The influence of intermittent negative pressure on the S. epidermidis growth showed notably lower median CFU with the interval therapy every hour compared to the S. aureus control group. This study contributes valuable insights into NPWT’s influence on the bacterial load, emphasizing the need for further research to reformulate its role in managing contaminated wounds.

Rapid and sensitive bioburden detection is of paramount importance in different applications including public health, and food and water safety. To overcome the traditional limitations of bacterial detection i.e., lengthy culture time, and complicated procedure, a low-cost, portable multichannel fluorometer coupled with machine learning (ML) has been implemented in this study. Five different strains of bacterial samples were tested along with the negative control for time-series fluorescence data collection and analysis. We applied different conventional unsupervised and supervised machine learning techniques with extracted features followed by preprocessing of the data. Initially, machine learning algorithms were applied for the qualitative detection of bacteria by binary classification followed by regression analysis to predict the level of contamination for E. coli. The multiclass classification was used to identify gram-positive, and gram-negative bacterial strains and differentiate all the bacterial strains tested. Our results show that around 97.9% accuracy can be achieved for bacterial contamination detection for as low as 1 CFU/mL while 92.1% accuracy can be achieved for differentiating the gram-positive and gram-negative strains. Additionally, with 1 minute of data, high accuracy is obtained for detecting bioburden, proving the multichannel fluorometer’s rapid detection capability. The multichannel fluorometer integrated with ML analytics is capable of automating data analysis and determining accurate and rapid bacterial detection on-site with the prediction of bioburden levels and differentiating bacterial strains and the protocol can be applied to the biosensors with a similar data type.

Abstract Introduction Infections due to extended spectrum beta-lactamase producing organisms (ESBL) have emerged as the leading cause of sepsis among hospitalized neonates in Botswana and much of sub-Saharan Africa and south Asia. Yet, ESBL reservoirs and transmission dynamics within the neonatal intensive care unit (NICU) environment are not well-understood. This study aimed to assess the efficiency and feasibility of a chromogenic-culture-media-based environmental sampling approach to characterize the ESBL bioburden within a NICU. Methods A series of four point-prevalence surveys were conducted at a 36-bed NICU at a public tertiary referral hospital in Botswana from January-June 2021. Samples were collected on 4 occasions under semi-sterile technique using 1) flocked swabs & templates (flat surfaces); 2) sterile syringe & tubing (water aspiration); and 3) structured swabbing techniques (hands & equipment). Swabs were transported in physiological saline-containing tubes, vortexed, and 10 µL was inoculated onto chromogenic-agar that was selective and differential for ESBL (CHROMagar™ ESBL, Paris, France), and streaking plates to isolate individual colonies. Bacterial colonies were quantified and phenotypically characterized using biochemical identification tests. Results In total, 567 samples were collected, 248 (44%) of which grew ESBL. Dense and consistent ESBL contamination was detected in and around sinks and certain high-touch surfaces, while transient contamination was demonstrated on medical equipment, caregivers/healthcare worker hands, insects, and feeding stations (including formula powder). Results were available within 24–72 h of collection. To collect, plate, and analyse 50 samples, we estimated a total expenditure of $269.40 USD for materials and 13.5 cumulative work hours among all personnel. Conclusions Using basic environmental sampling and laboratory techniques aided by chromogenic culture media, we identified ESBL reservoirs (sinks) and plausible transmission vehicles (medical equipment, infant formula, hands of caregivers/healthcare workers, & insects) in this NICU environment. This strategy was a simple and cost-efficient method to assess ESBL bioburden and may be feasible for use in other settings to support ongoing infection control assessments and outbreak investigations.

Microbial contamination in the hospital environment is a major concern for public health, since it significantly contributes to the onset of healthcare-associated infections (HAIs), which are further complicated by the alarming level of antimicrobial resistance (AMR) of HAI-associated pathogens. Chemical disinfection to control bioburden has a temporary effect and can favor the selection of resistant pathogens, as observed during the COVID-19 pandemic. Instead, probiotic-based sanitation (probiotic cleaning hygiene system, PCHS) was reported to stably abate pathogens, AMR, and HAIs. PCHS action is not rapid nor specific, being based on competitive exclusion, but the addition of lytic bacteriophages that quickly and specifically kill selected bacteria was shown to improve PCHS effectiveness. This study aimed to investigate the effect of such combined probiotic–phage sanitation (PCHSφ) in two Italian hospitals, targeting staphylococcal contamination. The results showed that PCHSφ could provide a significantly higher removal of staphylococci, including resistant strains, compared with disinfectants (−76%, p < 0.05) and PCHS alone (−50%, p < 0.05). Extraordinary sporadic chlorine disinfection appeared compatible with PCHSφ, while frequent routine chlorine usage inactivated the probiotic/phage components, preventing PCHSφ action. The collected data highlight the potential of a biological sanitation for better control of the infectious risk in healthcare facilities, without worsening pollution and AMR concerns.

The current communication seeks to provide an updated narrative review on latest methods of reducing implant contaminations used during spine surgery. Recent literature review has shown that both preoperative reprocessing and intraoperative handling of implants seem to contaminate implants. In brief, during preoperative phase, the implants undergo repeated bulk cleaning with dirty instruments from the OR, leading to residue buildup at the interfaces and possibly on the surfaces too. This, due to its concealed nature, remains unnoticed by the SPD (sterile processing department) or other hospital staff. Nevertheless, these can be avoided by using individually prepackaged presterilized implants. In the intraoperative phase, the implants (in the sterile field) are directly touched by the scrub tech with soiled (assisting the surgeon dispose the tissues from the instruments in use) gloves for loading onto an insertion device. It is then kept exposed on the working table (either separately or next to the used instruments as the pedicles hole are being prepared). Latest investigation has shown that by the time it is implanted in the patient, it can harbor up to 10e7 bacterial colony-forming units. The same implants were devoid of such colony-forming units, when sheathed by an impermeable sterile sheath around the sterile implant.

Background: The microbial contamination of a respirator can be evaluated through a count of the number of bacteria living on a non-sterilized surface (bioburden). This preliminary study investigated the external contamination of two different FFP2s over time by studying the bioburden values in increasing exposure times. Methods: FFP2 respirators of two different brands were used during routine clinical settings and examined through the bioburden test; for each brand, three devices were tested at 8, 16, and 30 h. Results: No significant differences were observed between mask brands (p = 0.113). There were only significant CFU differences between each mask and its control (p = 0.027 and p = 0.004). Conclusions: Both brands of respirators were found to be contaminated and this contamination increased with the increase in exposure time. Further studies are needed to investigate the exact amount of contamination that could be considered acceptable before discarding each used mask.

Caroline E Fife,1 Raphael A Yaakov,2 Thomas E Serena2 1CHI St. Luke’s Health, Wound Care Clinic, The Woodlands, TX, USA; 2SerenaGroup, Clinical Research, Cambridge, MA, USA Abstract: Epidermolysis bullosa (EB), often referred to as the butterfly disease, is a group of rare genetic conditions characterized by skin that is delicate and fragile as butterfly wings. The skin blisters in response to friction, minor injury, or trauma. In certain types of EB, other organs, such as the esophagus, can also be affected, and secondary complications may require multiple interventions. While there has been significant progress in classifying the disease – identifying genes and proteins involved – there have been few advances in the treatment of the disease. The care of the EB patient focuses on management of symptoms, protecting the skin, and preventing complications. In this case report, the use of a multivalent wound-healing ointment (Terrasil®) was evaluated in a 60-year-old patient with a history of junctional EB. A polymerase chain reaction-based culturing was utilized to quantitatively test for bacteria and fungi at baseline and follow-up visits. Pain assessment and wound area were also documented at each visit. Following the application of the wound care ointment, there was a reduction in wound surface area on central (96%) and distal mid-back (92%) by treatment visit three, and there was a 96% reduction on the left shoulder blade ulcer by treatment visit four. Moreover, there was a noticeable drop in the percentage of bacteria detected by polymerase chain reaction. The wound care ointment was also effective in eliminating the fungal species and reducing pain, itching, blistering, and cracking around the wound. Keywords: mechanobullous disorders, adjunctive therapy, bioburden, therapeutic management

Abstract Background Expressed human milk (EHM) feed preparation areas represent a potential source of unintentional nosocomial infection. Daily disinfection of environmental surfaces remains an essential intervention to mitigate nosocomial infections. The inefficiency of conventional cleaning and disinfection contributes to an increased risk for the acquisition of multi-drug resistant pathogens. “Non touch” technologies such as the pulsed xenon ultraviolet (PX-UVD) light device have documented sustained reduction in surface bacterial colonization and reduced cross contamination. Methods The impact of a PX-UVD on surface colony forming units per square centimeter (cfu/cm2) in feed preparation areas was evaluated following its implementation as standard care. A quasi-experimental study was performed documenting bacterial colonization from 6 high risk feed preparation areas in a community care hospital in South Africa. Pre and post conventional cleaning neutralizing rinse swabs were collected fortnightly over a 16 week control period prior to the introduction of the PX-UVD and compared to a matching set of samples for the PX-UVD period. Results A 90% reduction in total surface bioburden was noted from the control period (544 cfu/cm2) compared to the corresponding PX-UVD period (50 cfu/cm2). Sub -analysis of both the Pre-clean Control: Pre-clean PX-UVD counts as well as the Post-clean Control: Post-clean PX-UVD counts noted significant improvements (p

Sampling campaigns indoors have shown that occupants exposed to contaminated air generally exhibit diverse health outcomes. This study intends to assess the deposition rates of total settleable dust and bioburden in the indoor air of dwellings onto quartz fiber filters and electrostatic dust collectors (EDCs), respectively. EDC extracts were inoculated onto malt extract agar (MEA) and dichloran glycerol (DG18) agar-based media used for fungal contamination characterization, while tryptic soy agar (TSA) was applied for total bacteria assessment, and violet red bile agar (VRBA) for Gram-negative bacteria. Azole-resistance screening and molecular detection by qPCR was also performed. Dust loading rates ranged from 0.111 to 3.52, averaging 0.675 μg cm−2 day−1. Bacterial counts ranged from undetectable to 16.3 colony-forming units (CFU) m−2 day−1 and to 2.95 CFU m−2 day−1 in TSA and VRBA, respectively. Fungal contamination ranged from 1.97 to 35.4 CFU m−2 day−1 in MEA, and from undetectable to 48.8 CFU m−2 day−1 in DG18. Penicillium sp. presented the highest prevalence in MEA media (36.2%) and Cladosporium sp. in DG18 (39.2%). It was possible to observe: (a) settleable dust loadings and fungal contamination higher in dwellings with pets; (b) fungal species considered indicators of harmful fungal contamination; (c) Aspergillus section Candidi identified in supplemented media with voriconazole and posaconazole; (d) specific housing typologies and (e) specific housing characteristics influencing the microbial contamination.

Monitoring campaigns in several buildings have shown that occupants exposed to contaminated indoor air generally exhibit diverse health symptoms. This study intends to assess settleable dust loading rates and bioburden in Portuguese dwellings by passive sampling onto quartz fiber filters and electrostatic dust cloths (EDCs), respectively. Settled dust collected by EDCs was analyzed by culture-based methods (including azole-resistance screening) and qPCR, targeting four different toxigenic Aspergillus sections (Flavi, Fumigati, Circumdati, and Nidulantes). Dust loading rates and bioburden showed higher variability in the summer season. In both seasons, Penicillium sp. was the one with the highest prevalence (59.1% winter; 58.1% summer), followed by Aspergillus sp. in winter (13.0%). Fungal contamination increased in the winter period, while bacterial counts decreased. Aspergillus sections Circumdati and Nidulantes, detected in voriconazole supplemented media, and Aspergillus sections Fumigati and Nidulantes, detected by molecular tools, were found in the winter samples. This study reinforces the importance of applying: (a) Passive sampling methods in campaigns in dwellings; (b) two different culture media (MEA and DG18) to assess fungi; (c) in parallel, molecular tools targeting the most suitable indicators of fungal contamination; and (d) azole resistance screening to unveil azole resistance detection in fungal species.

The exposure to particles and bioaerosols has been associated with the increase in health effects in children. The objective of this study was to assess the indoor exposure to bioburden in the indoor microenvironments more frequented by children. Air particulate matter (PM) and settled dust were sampled in 33 dwellings and four schools with a medium volume sampler and with a passive method using electrostatic dust collectors (EDC), respectively. Settled dust collected by EDC was analyzed by culture-based methods (including azole resistance profile) and using qPCR. Results showed that the PM2.5 and PM10 concentrations in classrooms (31.15 μg/m3 and 57.83 μg/m3, respectively) were higher than in homes (15.26 μg/m3 and 18.95 μg/m3, respectively) and highly exceeded the limit values established by the Portuguese legislation for indoor air quality. The fungal species most commonly found in bedrooms was Penicillium sp. (91.79%), whereas, in living rooms, it was Rhizopus sp. (37.95%). Aspergillus sections with toxigenic potential were found in bedrooms and living rooms and were able to grow on VOR. Although not correlated with PM, EDC provided information regarding the bioburden. Future studies, applying EDC coupled with PM assessment, should be implemented to allow for a long-term integrated sample of organic dust.

The assessment and control of microbial contamination in health care facilities is presently a mandatory and vital part of strategies to prevent and control hospital-acquired infections. This study aims to assess the bioburden with two passive sampling methods (30 ventilations grids swabs and 16 electrostatic dust collectors (EDCs)) at Clinical Pathology Services. The fungal burden was characterized through molecular tools, antifungal resistance, and the mycotoxins and cytotoxicity profile. Total bacteria presented the highest prevalence in both matrixes, whereas Gram-bacteria presented the lowest. Swabs presented a higher prevalence (27.6%) for fungal burden. Chrysonilia sitophila presented the highest prevalence in swabs, whereas for EDCs, C. sitophila and Mucor sp. were the most prevalent. Concerning Aspergillus genera on swabs, section Flavi was the one with the highest prevalence (58.02%), whereas, for EDCs, section Versicolores was the only section observed (100%). Aspergillus section Fumigati was detected in 10 swabs and 7 EDC samples and Aspergillus section Versicolores was detected in one EDC sample. Fungal growth on azole-supplemented media was observed in eight EDC samples. No mycotoxins were detected in any of the samples. A low cytotoxic effect was observed in two sites upon incubation of collected samples with A549 and SK cells and in two other sites upon incubation of collected samples with SK cells only. A medium cytotoxic effect was observed with one EDC sample upon incubation with A549 cells. This study reinforces the need of determination of the azole resistance profile for fungal species and allowed a preliminary risk characterization regarding the cytotoxicity. An intervention including the use of a ultraviolet with wavelength between 200 nm and 280 nm (UVC)—emitting device and an increased maintenance and cleaning of the central heating, ventilation, and air conditioning (HVAC) systems should be ensured to promote the reduction of microbial contamination.

A sporicidal agent formula based on a mixture of peroxyacetic acid and hydrogen peroxide was assessed for its efficacy on a representative sample of vinyl surface material. Vinyl is the construction material of wall and floor lining in pharmaceutical plants. The experimental manipulations; applied herein, simulated the actual biocidal agent preparation and were carried out using USP purified water, test temperature was 20–25 °C, RH% was 40–60% and pH was 3.08 and 2.86 for 1% and 2% (v/v) respectively. Following the selection of the optimum method of antimicrobial activity neutralization, two disinfectant concentrations were examined for their sporicidal activity. The results of carrier test revealed that the disinfectant concentration (2%) (v/v) was significantly effective as a sporicidal agent after 5 and 10 min for Aspergillus brasiliensis and Bacillus subtilis subsp. spizizenii, respectively, while the concentration of 1% (v/v) did not achieve even one logarithmic reduction after 20 min. The agent was able to achieve more than 100 times reduction from the initial bioburden on the surfaces when used in the concentration of 2% (v/v) after 10 min of contact time. The ideal kinetics of microbial death usually follows 2 parts (by averaging the responses with time for the 3 replicates): initial slow rate of death followed by higher rate. The initial sigmoidal part was only observed with B. subtilis upon exposure to 2% (v/v) sporicidal agent. Elimination time for B. subtilis spores was 15 min which was about double the time required for eradication of A. brasiliensis.

Ventilator associated pneumonia (VAP) remains a serious problem for critically ill patients. We swabbed nine reusable areas on 20 clinically-used ventilators from a VA Hospital shortly after they had been removed from patients and identified bacterial isolates. No bacteria were isolated from most of the samples and of the samples that did grow bacteria, the majority of those had fewer than 10 colonies. The bacteria that were isolated were primarily non-pathogenic Gram-positive skin flora. Of the 20 ventilators swabbed, only one cultured bacteria associated with nosocomial infections: methicillin-resistant S.aureus. The most commonly contaminated areas were those most likely to be touched by healthcare professionals: the power button and the screen. The areas in closest proximity to the patients, the inspiratory and expiratory ports were the least often contaminated areas. Overall, very few bacteria were transferred to the reusable areas of the ventilators following clinical use.

The research has been carried out on the microbial hazards by using contaminated surgical instruments. In the operating room medical devices that have contact with sterile body tissues or fluids are considered as dangerous surgical tools. These surgical tools should be sterile when used because any microbial contamination could result in disease transmission and would leads to high number of Surgical Site Infections (SSI). The aim of this study is to establish the possible presence of known bacterial pathogens on surgical instruments in hospital settings, and to evaluate their antibiotic susceptibility profile. For this purpose a comparative research was conducted; the 40 samples were collected from instruments at the start and end of the procedure with the sterile cotton swabs; from the Gynea department, Abbasi Shaheed Hospital. These samples were cultured on the selective and differential media and incubated at 37°C for 24hr. Next day, these samples were processed and identified in Microbiology laboratory as per standard microbiological techniques; including the microscopy, biochemical tests, enzymes test and other culture characterizations test. The isolated microorganisms include E.coli, Staph aureus, Staph. saprophyticus, Strep viridians, Strep. bovis, Bacillus subtilis, and Pseudomonas spp. These microorganism were isolated from both autoclaved and non-autoclaved pre and post-operative surgical instruments respectively. Antibiotic susceptibility test was carried out by Kirby Bauer Method. Meropenum found to be the most effective against E.coli, Gentamycin and Novobiocin shows activity against S.aureus, Erythromycin also give zone of inhibition against S.bovis, Vancomycin shows activity against S.viridans, where Cephalenin and Ampicillin were also found to be effective against S. saprophyticus. Surgical instruments should be regarded as a possible source of nosocomial infection s since bacteria from them can be carried from the hands of theatre personnel to the patient undergoing surgery or through re-dispersed bacteria from surfaces during surgery. Therefore surgical procedure to prevent cross contamination instruments are sterilized and decontaminated. There has been advance in Surgical Site Infection (SSI) control practices which involves improved operating room ventilation, sterilization methods, surgical methods and availability of antimicrobial prophylaxis.

Ventilator associated pneumonia (VAP) remains a serious problem for critically ill patients. We swabbed nine reusable areas on 20 clinically-used ventilators from a VA Hospital shortly after they had been removed from patients and identified bacterial isolates. No bacteria were isolated from most of the samples and of the samples that did grow bacteria, the majority of those had fewer than 10 colonies. The bacteria that were isolated were primarily non-pathogenic Gram-positive skin flora. Of the 20 ventilators swabbed, only one cultured bacteria associated with nosocomial infections: methicillin-resistant S.aureus. The most commonly contaminated areas were those most likely to be touched by healthcare professionals: the power button and the screen. The areas in closest proximity to the patients, the inspiratory and expiratory ports were the least often contaminated areas. Overall, very few bacteria were transferred to the reusable areas of the ventilators following clinical use.

Background and Objectives: Paper and paperboard packaging play an important role in safety and quality of food products. Common bacteria of paper and paperboard food packaging could grow due to specific conditions included humidity, tem- perature and major nutrition to contaminate the food. The purpose of this research was to investigate numbers and the types of bacteria in the food packaging paperboard. Materials and Methods: The surface and the depth of the each paperboard sample were examined by the dimension of one cm2 and one gram. The paperboard samples were randomly collected from popular confectionaries and fast food restaurants in Tehran, Iran. Results: The results indicated the range of 0.2×103 to >1.0×105 cfu/1g bacterial contamination in paperboard food packaging. Also, most detected bacteria were from spore forming and family Bacillaceae. Conclusion: The bioburden paperboard used for food packaging showed high contamination rate more than standard accep- tance level.

Organic dust is widespread in the environment including occupational settings, such as bakeries. Recently, a new collection device—the electrostatic dust cloth (EDC)—has been described for the assessment of occupational exposures. The aim of this study was to investigate the suitability of EDC for identifying the distribution patterns and exposure concentrations of particulate matter and microbial contaminants such as fungi and bacteria in bakeries. Twelve bakeries were selected, and dust was allowed to settle for 13 to 16 days on EDCs (a total of 33 samples). Particle counts and size distribution (0.3 µm, 0.5 µm, 1 µm, 2.5 µm, 5 µm and 10 µm) were measured with direct-reading equipment. Higher EDC mass was significantly correlated (p values < 0.05) with higher fungal load on dichloran glycerol (DG18) and with particle size distribution in the 0.3 µm, 0.5 µm, 1.0 µm and 10.0 µm range. Fungal levels on malt extract agar (MEA) ranged from 0 to 2886 CFU/m2 EDC in the warehouse setting, 0 to 500 CFU/m2 EDC in the production setting, and 0 to 3135 CFU/m2 EDC in the store. Penicillium sp. (42.56%) was the most frequent fungi. Total bacterial load ranged from 0 to 18,859 CFU/m2 EDC in the warehouse, 0 to 71,656 CFU/m2 EDC in production, and 0 to 21,746 CFU/m2 EDC in the store. EDC assessment provided a longer-term integrated sample of organic dust, useful for identifying critical worksites in which particulate matter and bio-burden exposures are elevated. These findings suggest that EDC can be applied as a screening method for particulate matter-exposure assessment and as a complementary method to quantify exposures in occupational environments.