Megan Hargreaves

Characterization of expiration air jets and droplet size distributions immediately at the mouth opening

Abstract Size distributions of expiratory droplets expelled during coughing and speaking and the velocities of the expiration air jets of healthy volunteers were measured. Droplet size was measured using the interferometric Mie imaging (IMI) technique while the particle image velocimetry (PIV) technique was used for measuring air velocity. These techniques allowed measurements in close proximity to the mouth and avoided air sampling losses. The average expiration air velocity was 11.7m/s for coughing and 3.9m/s for speaking. Under the experimental setting, evaporation and condensation effects had negligible impact on the measured droplet size. The geometric mean diameter of droplets from coughing was 13.5μm and it was 16.0μm for speaking (counting 1–100). The estimated total number of droplets expelled ranged from 947 to 2085 per cough and 112–6720 for speaking. The estimated droplet concentrations for coughing ranged from 2.4 to 5.2cm−3 per cough and 0.004–0.223cm−3 for speaking.

A pilot investigation into associations between indoor airborne fungal and non-biological particle concentrations in residential houses in Brisbane, Australia

Indoor air contains a complex mixture of bioaerosols such as fungi, bacteria and allergens, as well as non-biological particles including products from various combustion processes. To date little work has been done to investigate the interactions and associations between particles of biological and non-biological origin, however, any occurring interactions could affect pollutant behaviour in the air and ultimately the effect they have on health. The aim of this work was to examine associations between the concentration levels of airborne particles and fungi measured in 14 residential suburban houses in Brisbane. The most frequently isolated fungal genus was Cladosporium, Curvularia, Alternaria, Fusarium and Penicillium. The average outdoor and indoor (living room) concentrations of fungal colony forming units were 1133+/-759 and 810+/-389, respectively. Average outdoor and indoor (normal ventilation) concentrations of submicrometre and supermicrometre particles were 23.8 x 10(3) and 21.7 x 10(3) (particles/cm(3)), 1.78 and 1.74 (particles/cm(3)), respectively. The study showed that no statistically significant associations between the fungal spore and submicrometre particle concentrations or PM(2.5) were present, while a weak but statistically significant relationship was found between fungal and supermicrometre particle concentrations (for the outdoors R(2)=0.4, P=0.03 and for a living room R(2)=0.3, P=0.04). A similarity in behaviour between the submicrometre particle and fungal spore concentrations was that the fungal spore concentrations were related directly to the distance from the source (a nearby park), in a very similar way in which the submicrometre particles originating from vehicle emissions from a road, were dependent on the distance to the road. In the immediate proximity to the park, fungal concentrations rose up to approximately 3100 CFU/m(3), whereas for houses more than 150 m away from the park the concentrations of fungi were below 1000 CFU/m(3). Recommendations have been provided as the future study designs to gain a deeper insight into the relationships between biological and non-biological particles.

Isolation of Nontuberculous Mycobacteria (NTM) from Household Water and Shower Aerosols in Patients with Pulmonary Disease Caused by NTM

ABSTRACT It has been postulated that susceptible individuals may acquire infection with nontuberculous mycobacteria (NTM) from water and aerosol exposure. This study examined household water and shower aerosols of patients with NTM pulmonary disease. The mycobacteria isolated from clinical samples from 20 patients included M. avium (5 patients), M. intracellulare (12 patients), M. abscessus (7 patients), M. gordonae (1 patient), M. lentiflavum (1 patient), M. fortuitum (1 patient), M. peregrinum (1 patient), M. chelonae (1 patient), M. triplex (1 patient), and M. kansasii (1 patient). One-liter water samples and swabs were collected from all taps, and swimming pools or rainwater tanks. Shower aerosols were sampled using Andersen six-stage cascade impactors. For a subgroup of patients, real-time PCR was performed and high-resolution melt profiles were compared to those of ATCC control strains. Pathogenic mycobacteria were isolated from 19 homes. Species identified in the home matched that found in the patient in seven (35%) cases: M. abscessus (3 cases), M. avium (1 case), M. gordonae (1 case), M. lentiflavum (1 case), and M. kansasii (1 case). In an additional patient with M. abscessus infection, this species was isolated from potable water supplying her home. NTM grown from aerosols included M. abscessus (3 homes), M. gordonae (2 homes), M. kansasii (1 home), M. fortuitum complex (4 homes), M. mucogenicum (1 home), and M. wolinskyi (1 home). NTM causing human disease can be isolated from household water and aerosols. The evidence appears strongest for M. avium, M. kansasii, M. lentiflavum, and M. abscessus. Despite a predominance of disease due to M. intracellulare, we found no evidence for acquisition of infection from household water for this species.

Real-time measurement of bacterial aerosols with the UVAPS: performance evaluation

The Ultraviolet Aerodynamic Particle Sizer (UVAPS, Model 3312, TSI Inc., St. Paul, MN) spectrometer is the only commercially available aerosol counter for real-time monitoring of viable bioaerosols. Though the feasibility of this technique to monitor bioaerosols has been previously demonstrated by the instrument designers in a number of studies, the collection of meaningful data and their correct interpretation are still not possible without a thorough understanding of its capabilities and limitations. This paper presents the results of the first independent study aimed towards evaluating selectivity, sensitivity, counting efficiency, and the detection limits of the UVAPS. The study has demonstrated limitations in the capability of the instrument to measure bacterial spores that is explained by biochemical composition of the spores, which contain only minute amounts of the specific fluorophores that appeared to be below the instrument sensitivity level. The results were also indicative of strong sensitivity of the UVAPS to the physiological state of bacteria. Counting efficiency of the fluorescent particles was shown to depend on particle concentration with the upper limit of detection of the UVAPS around 6 x 107 particles/ m3.

Performance evaluation of the UVAPS: Influence of physiological age of airborne bacteria and bacterial stress

This study evaluated the effect of bacterial physiology, such as physiological age and stress, on the performance of the ultraviolet aerodynamic particle sizer (UV-APS, model 3312, TSI Inc., St. Paul, MN). Intensity of the fluorescent signals was measured for three bacteria having various sensitivities to environmental stresses, Bacillus subtilus (spores and vegetative cells), Pseudomonas fluorescens, and Micrococcus luteus. The performance of the UVAPS was found to depend on the type of airborne bacteria. In addition, the fluorescence signals for stationary-phase bacteria were generally stronger than for their log-phase counterparts. These results indicated that bacterial injury due to environmental stresses has a strong influence on the measured fluorescence signals. This hypothesis was confirmed by obtaining a linear relationship between the percentage of fluorescent particles and the proportion of injured bacteria in the total population of cultivable bacteria in samples simultaneously collected with the AGI-30 impingers. This indicates that the amount of fluorophors (specifically NADH) within injured bacteria is below the UVAPS sensitivity level. The practical implications of these findings are discussed in the paper. The reported results contribute to broadening our understanding of the method and may assist in developing sampling strategies for the application of the UVAPS to various bioaerosol studies.

Factors associated with the isolation of Nontuberculous mycobacteria (NTM) from a large municipal water system in Brisbane, Australia

BackgroundNontuberculous mycobacteria (NTM) are normal inhabitants of a variety of environmental reservoirs including natural and municipal water. The aim of this study was to document the variety of species of NTM in potable water in Brisbane, QLD, with a specific interest in the main pathogens responsible for disease in this region and to explore factors associated with the isolation of NTM. One-litre water samples were collected from 189 routine collection sites in summer and 195 sites in winter. Samples were split, with half decontaminated with CPC 0.005%, then concentrated by filtration and cultured on 7H11 plates in MGIT tubes (winter only).ResultsMycobacteria were grown from 40.21% sites in Summer (76/189) and 82.05% sites in winter (160/195). The winter samples yielded the greatest number and variety of mycobacteria as there was a high degree of subculture overgrowth and contamination in summer. Of those samples that did yield mycobacteria in summer, the variety of species differed from those isolated in winter. The inclusion of liquid media increased the yield for some species of NTM. Species that have been documented to cause disease in humans residing in Brisbane that were also found in water include M. gordonae, M. kansasii, M. abscessus, M. chelonae, M. fortuitum complex, M. intracellulare, M. avium complex, M. flavescens, M. interjectum, M. lentiflavum, M. mucogenicum, M. simiae, M. szulgai, M. terrae. M. kansasii was frequently isolated, but M. avium and M. intracellulare (the main pathogens responsible for disease is QLD) were isolated infrequently. Distance of sampling site from treatment plant in summer was associated with isolation of NTM. Pathogenic NTM (defined as those known to cause disease in QLD) were more likely to be identified from sites with narrower diameter pipes, predominantly distribution sample points, and from sites with asbestos cement or modified PVC pipes.ConclusionsNTM responsible for human disease can be found in large urban water distribution systems in Australia. Based on our findings, additional point chlorination, maintenance of more constant pressure gradients in the system, and the utilisation of particular pipe materials should be considered.

Mycobacterium abscessus isolated from municipal water - a potential source of human infection

BackgroundMycobacterium abscessus is a rapidly growing mycobacterium responsible for progressive pulmonary disease, soft tissue and wound infections. The incidence of disease due to M. abscessus has been increasing in Queensland. In a study of Brisbane drinking water, M. abscessus was isolated from ten different locations.The aim of this study was to compare genotypically the M. abscessus isolates obtained from water to those obtained from human clinical specimens.MethodsBetween 2007 and 2009, eleven isolates confirmed as M. abscessus were recovered from potable water, one strain was isolated from a rainwater tank and another from a swimming pool and two from domestic taps. Seventy-four clinical isolates referred during the same time period were available for comparison using rep-PCR strain typing (Diversilab).ResultsThe drinking water isolates formed two clusters with ≥97% genetic similarity (Water patterns 1 and 2). The tankwater isolate (WP4), one municipal water isolate (WP3) and the pool isolate (WP5) were distinctly different. Patient isolates formed clusters with all of the water isolates except for WP3. Further patient isolates were unrelated to the water isolates.ConclusionThe high degree of similarity between strains of M. abscessus from potable water and strains causing infection in humans from the same geographical area, strengthens the possibility that drinking water may be the source of infection in these patients.

Sourcing faecal pollution from onsite wastewater treatment systems in surface waters using antibiotic resistance analysis

Aims:  To identify the sources of faecal contamination in investigated surface waters and to determine the significance of onsite wastewater treatment systems (OWTS) as a major contributor to faecal contamination.

Antibiotic resistance and virulence traits in clinical and environmental Enterococcus faecalis and Enterococcus faecium isolates.

This study compared virulence and antibiotic resistance traits in clinical and environmental Enterococcus faecalis and Enterococcus faecium isolates. E. faecalis isolates harboured a broader spectrum of virulence determinants compared to E. faecium isolates. The virulence traits Cyl-A, Cyl-B, Cyl-M, gel-E, esp and acm were tested and environmental isolates predominantly harboured gel-E (80% of E. faecalis and 31.9% of E. faecium) whereas esp was more prevalent in clinical isolates (67.8% of E. faecalis and 70.4% of E. faecium). E. faecalis and E. faecium isolated from water had different antibiotic resistance patterns compared to those isolated from clinical samples. Linezolid resistance was not observed in any isolates tested and vancomycin resistance was observed only in clinical isolates. Resistance to other antibiotics (tetracycline, gentamicin, ciprofloxacin and ampicillin) was detected in both clinical and water isolates. Clinical isolates were more resistant to all the antibiotics tested compared to water isolates. Multi-drug resistance was more prevalent in clinical isolates (71.2% of E. faecalis and 70.3% of E. faecium) compared to water isolates (only 5.7% E. faecium). tet L and tet M genes were predominantly identified in tetracycline-resistant isolates. All water and clinical isolates resistant to ciprofloxacin and ampicillin contained mutations in the gyrA, parC and pbp5 genes. A significant correlation was found between the presence of virulence determinants and antibiotic resistance in all the isolates tested in this study (p<0.05). The presence of antibiotic resistant enterococci, together with associated virulence traits, in surface recreational water could be a public health risk.

Mycobacterium lentiflavum in Drinking Water Supplies, Australia

Humans may acquire infection from potable water.