Evaluation of a new culture medium for isolation of nontuberculous mycobacteria from environmental water samples.

Nontuberculous mycobacteria (NTM) are waterborne pathogens commonly found in building water systems where they are a primary concern to vulnerable patient populations and can cause severe disease. The recovery of NTM from environmental samples can be a laborious undertaking and current pre-treatment methods and selective media lack sensitivity. We explored the use of the highly selective Rapidly Growing Mycobacteria (RGM) medium for culturing NTM from environmental water samples compared to existing methods. In total, 223 environmental water samples, including potable and non-potable water, were cultured for NTM using three culture media. In addition to direct culture on RGM medium, each sample was cultured on Middlebrook 7H10 medium and Mitchison 7H11 medium after pre-treatment with 0.2M KCl-HCl. Additionally, 33 distinct species of NTM were inoculated onto RGM medium and 7H10 medium in parallel to directly compare their growth. The use of RGM medium alone without pre-treatment provided a sensitivity (91%) comparable to that offered by culture on both 7H10 and 7H11 with acid pretreatment (combined sensitivity; 86%) with significantly less overgrowth and interference from other organisms on RGM medium. The average concentration of NTM observed on RGM medium alone was comparable to or greater than the NTM concentration on either medium alone or combined. Thirty-three species were examined in parallel and all tested strains of 27 of these species successfully grew on RGM medium, including 19 of 21 from the CDC's healthcare-associated infections species list. RGM medium was successful at recovering environmental NTM without a pre-treatment, greatly reducing labor and materials required to process samples. Simplification of culture processing for environmental NTM will allow for a better assessment of their presence in building water systems and the potential for reduced exposure of susceptible populations.

Effects of volume, velocity, and composition on the resistance to synthetic blood penetration of N95 filtering facepiece respirators and other head/facial personal protective equipment.

Surgical N95 filtering facepiece respirators (surgical N95 FFRs) are National Institute for Occupational Safety and Health-approved N95 filtering facepiece respirators (N95 FFRs) cleared by the Food and Drug Administration for resistance to liquid penetration and flammability. A recent study showed that several N95 FFR models performed as well as surgical N95 FFRs in synthetic blood penetration tests that evaluate resistance to penetration by horizontal projection. This aspect, in addition to the influence of other factors on liquid penetration, are not well studied. To address this issue, the effect of liquid volume (1 mL and 2 mL), spray velocity (450 cm/sec and 635 cm/sec), and liquid composition (synthetic blood and diluted synthetic blood) were evaluated. Four types of common protective devices were studied: N95 FFRs, surgical N95 FFRs, surgical masks, and powered air-purifying respirator (PAPR) hoods. For each protective device type, five models were analyzed using a protocol based on the F1862 ASTM International (2017) test method. Reduced liquid volume had a significant effect in only 3 of 20 models. Increased velocity had significantly greater penetration in 9 of 20 models. Diluted synthetic blood had significantly more penetration in 8 of 20 models. This last result was not expected because, in hydrostatic tests, surface tension of the diluted blood would be expected to reduce penetrability; however, across all models tested, data showed that the diluted spray was more penetrable. The study results suggest that fluid composition may be as important as velocity when considering liquid spray penetration. Furthermore, the penetrability of a spray may be inversely related to the penetrability through direct hydrostatic contact.

A new approach to measure the resistance of fabric to liquid and viral penetration.

Protective clothing manufacturers routinely test their products for resistance to liquid and viral penetration. Several of the test methods specified by the American Society for Testing and Materials (ASTM) and the International Organization for Standardization (ISO) for penetration testing produce binary results (i.e. pass or fail), deliver imprecise pressure regulation, and do not record the location at which penetration events occur. Instead, our approach measures a continuous variable (time of penetration) during a slow and continuous increase of hydrostatic pressure and retains the location of penetration events. Using a fluorescent dye to enhance visual detection, we evaluate temporal and spatial patterns of penetration events. We then compare the time of liquid penetration with the time of penetration of two bacteriophages (Phi-X174 and MS2). For the fabric tested, the mean viral penetration occurred 0.29 minutes earlier than liquid penetration when solved by logistic regression. The breakthrough time of MS2 was not different from the Phi-X174 bacteriophage. The time of liquid penetration was a latent indicator of the time of viral penetration.

Evaluation of Apparatus Used to Test Liquid through Protective Materials: Comparison of a Modified Dot-Blot Apparatus to the ASTM Penetration Cell.

Personal protective equipment (PPE), such as gowns used in the latest Ebola outbreak in Western Africa, are critical in preventing the spread of deadly diseases. Appropriate test systems and test soils are needed to adequately evaluate PPE. ASTM F903, Standard Test Method for Resistance of Materials Used in Protective Clothing to Penetration by Liquid, has been used for decades to test fabrics' resistance to liquid penetration. However, this test apparatus requires at least 60 mL of test solutions, is labor intensive, and has problems with leakage around the gaskets. We compared the F903 test apparatus to a modified dot-blot apparatus to evaluate the visual penetration of a blood test soil. A series of commercially available gowns and drapes were tested in each apparatus. Using blood test soil at 2 psi, there was no statistically significant difference between the two methods except for in one gown. By comparing this gown in the ASTM test apparatus with and without a screen, the particular screen selected did not account for the difference between the dot-blot and F903 apparatuses; however, it is conceivable that a particular screen/fabric combination could account for this difference. The modified dot-blot apparatus was evaluated using three different test solutions: blood, vomit, and a labeled protein (goat anti-rabbit immunoglobulin G-horseradish peroxidase [GaR IgG-HRP]) in a blood test soil solution. This testing revealed significant difference in penetration for some of the PPE garments. The modified dot-blot had several large advantages over the ASTM apparatus-over six times less specimen volume and no edge or gasket leakage. In addition, nitrocellulose can be easily incorporated into the modified dot-blot apparatus, enabling the trapping of viruses and proteins that penetrate PPE-thus permitting the use of antibodies to quickly and sensitively detect penetration.

Screening Heroin Smokers Attending Community Drug Services for COPD.

BACKGROUND: Heroin smoking is associated with deprivation, early onset severe emphysema, premature morbidity and mortality, and high use of health care, but individuals engage poorly with traditional health services. METHODS: In this cross-sectional study, we screened a population of heroin smokers, prescribed opiate substitution therapy by community drug services, for airway disease. We assessed drug exposure, respiratory symptoms, health status, and COPD prevalence. Subjects completed spirometry, completed Medical Research Council (MRC) Dyspnea Scale, COPD Assessment Tool (CAT) questionnaire, recorded drug exposure, and provided feedback. RESULTS: A total of 753 people (73% of those approached) completed screening, with 260 participants (35%) having COPD using FEV1/FVC < 0.7 and 293 (39%) participants having COPD using the lower limit of normal. A further 112 participants (15%) had asthma-COPD overlap (ACO) with features of COPD and asthma. Compared with those with normal spirometry, participants with COPD were more breathless (MRC score 3.1 vs 1.9; P < .001) and had worse health status (CAT score 22.9 vs 13.4; P < .001), respectively. Individuals with COPD had smoked cigarettes (P < .001), heroin (P < .001), and crack (P = .03) for longer and were more likely to still be smoking heroin (P < .01). Feedback was strongly positive, with 92% of respondents happy for other health-care appointments to be colocated with drug key worker appointments. CONCLUSIONS: Most heroin smokers had COPD or ACO, most commonly mild to moderate disease. In high-risk areas, screening this population provides an opportunity to reduce symptoms and risk. Anchoring respiratory health screening to drug center appointments delivers high completion and satisfaction and is an appropriate model for screening other hard-to-reach populations.

The Surface Tension of Synthetic Blood used for ASTM F1670 Penetration Tests.

The American Society for Testing and Material (ASTM International) F1670 test method was based on research involving transmission of bloodborne pathogens (Hepatitis B, Hepatitis C, and HIV) in the 1980s. The test method details the measurement of synthetic blood penetration through garments. A key parameter affecting penetration is synthetic blood surface tension which is measured via du Noüy ring tensiometer. However, little is known about the sources of variation impacting surface tension measurements. In this study, the synthetic blood used for ASTM F1670 was evaluated from within the ASTM F903 test apparatus and with two mixing treatments. Measurements were compared against two outside laboratories and with two alternate tensiometric methods (pendant drop and capillary rise). It was found that using the methods specified in the ASTM F1670 test method, surface tension of the synthetic blood was not 40-44 dynes/cm as was expected. The surface tension was initially above 50 dynes/cm and declined to below 40 dynes/cm after 60 minutes. The surface tension within the penetration cell was relatively constant over time, showing that the surface tension measurements outside the penetration cell are not indicative of the surface tension within the apparatus during the test. Shaking the synthetic blood, a mixing procedure detailed in the ASTM F1670 test method, increased the surface tension. The increase was greatest in a container having more airspace. Du Noüy ring measurements by NIOSH compared to external labs were within 15%. Testing with alternate methods showed that the "open-to-atmosphere" methods (ring and drop) began lower and declined rapidly when compared to the "closed-to-atmosphere" method (capillary). Results of this research will help amend the ASTM F1670 standard to better characterize the measurement and handling of synthetic blood used in the ASTM F1670 test and to provide a framework for consideration of test fluid used in future ASTM standards.

The size of lipid rafts: an atomic force microscopy study of ganglioside GM1 domains in sphingomyelin/DOPC/cholesterol membranes.

Atomic force microscopy has been used to study the distribution of ganglioside GM1 in model membranes composed of ternary lipid mixtures that mimic the composition of lipid rafts. The results demonstrate that addition of 1% GM1 to 1:1:1 sphingomyelin/dioleoylphosphatidylcholine/cholesterol monolayers leads to the formation of small ganglioside-rich microdomains (40-100 nm in size) that are localized preferentially in the more ordered sphingomyelin/cholesterol-rich phase. With 5% GM1 some GM1 microdomains are also detected in the dioleoylphosphatidylcholine-rich phase. A similar preferential localization of GM1 in the ordered phase is observed for bilayers with the same ternary lipid mixture in the upper leaflet. The small GM1-rich domains observed in these experiments are similar to the sizes for lipid rafts in natural membranes but considerably smaller than the ordered bilayer domains that have been shown to be enriched in GM1 in recent fluorescence microscopy studies of lipid bilayers. The combined data from a number of studies of model membranes indicate that lateral organization occurs on a variety of length scales and mimics many of the properties of natural membranes.