Influence of seasons and sites on bioaerosols in indoor wastewater treatment plants and proposal for air quality indicators.

During wastewater treatment, bioaerosols are generated and, can either remain in suspension for several hours or settle on surfaces and workers may be exposed. The presence of pathogens in the air could contribute to an increased frequency of gastrointestinal or respiratory illness amongst workers. Due to harsh winter conditions in Eastern Canada, many of the steps in the wastewater treatment process occur indoors, leading to a greater risk of significant occupational exposure especially if there is inadequate ventilation or a lack of personal protection. This work has used stationary sampling at various indoor wastewater treatment steps both in winter and summer. Bioaerosols were evaluated using both culture and molecular methods along with ventilation characterization. Endotoxins were quantified, as well as total cultivable and gram-negative bacteria and pathogen indicators using qPCR. This study highlights the presence of potential pathogens at all steps in the treatment process, which may represent a potential occupational hazard. Comparisons between summer and winter data suggest that water temperature is an important factor for microbial activity and suggest that increasing the rate of air changes per hour in summer would be beneficial to reduce the concentration of bioaerosols during this time of the year. The screening, grit/FOGs removal and biofiltration were the most bioaerosol-loaded sites. Based on strong correlations, we suggest the reconsideration of exposure limits in WWTPs. Workers should be encouraged to use personal respiratory protection to limit the risk of health problems, especially during long-term work.Implications: The work presented herein showcases significant correlations between concentrations of endotoxins, cultivable bacteria, gram-negative bacteria, and total bacteria by qPCR from air collected in indoor wastewater treatment plants. These correlations lead us to propose new limit of exposure values, revisited to fit the endotoxin exposure limits recommendations. The results can serve as guidelines for future proposals for air quality indicators.

Human viral pathogens are pervasive in wastewater treatment center aerosols.

Wastewater treatment center (WTC) workers may be vulnerable to diseases caused by viruses, such as the common cold, influenza and gastro-intestinal infections. Although there is a substantial body of literature characterizing the microbial community found in wastewater, only a few studies have characterized the viral component of WTC aerosols, despite the fact that most diseases affecting WTC workers are of viral origin and that some of these viruses are transmitted through the air. In this study, we evaluated in four WTCs the presence of 11 viral pathogens of particular concern in this milieu and used a metagenomic approach to characterize the total viral community in the air of one of those WTCs. The presence of viruses in aerosols in different locations of individual WTCs was evaluated and the results obtained with four commonly used air samplers were compared. We detected four of the eleven viruses tested, including human adenovirus (hAdV), rotavirus, hepatitis A virus (HAV) and Herpes Simplex virus type 1 (HSV1). The results of the metagenomic assay uncovered very few viral RNA sequences in WTC aerosols, however sequences from human DNA viruses were in much greater relative abundance.

Comparison of four ß-glucuronidase and ß-galactosidase-based commercial culture methods used to detect Escherichia coli and total coliforms in water.

The MI agar, Colilert(®), Chromocult coliform(®) agar, and DC with BCIG agar chromogenic culture-based methods used to assess microbiological quality of drinking water were compared in terms of their ubiquity, sensitivity, ease of use, growth of atypical colonies and affordability. For ubiquity, 129 total coliform (representing 76 species) and 19 Escherichia coli strains were tested. Then, 635 1-L well water samples were divided into 100 mL subsamples for testing by all four methods. Test results showed that 70.5, 52.7, 36.4, and 23.3% of the non-E. coli total coliform strains and 94.7, 94.7, 89.5, and 89.5% of the 19 E. coli strains yielded a positive signal with the four methods, respectively. They also yielded a total coliform positive signal for 66.5, 51.7, 64.9, and 55.0% and an E. coli positive signal for 16.1, 14.8, 17.3, and 13.4% of the 635 well water samples tested, respectively. Results showed that Colilert(®) is the most expensive method tested in terms of reactants, yet it is the easiest to use. Large numbers of atypical colonies were also often observed on Chromocult coliform(®) and DC with BCIG, thereby challenging the target microorganism count. Thus, the MI agar method seems to be the best option for the assessment of drinking water quality.

Multiparametric comparison of chromogenic-based culture methods used to assess the microbiological quality of drinking water and the mFC method combined with a molecular confirmation procedure.

MI agar and Colilert(®), as well as mFC agar combined with an Escherichia coli-specific molecular assay (mFC + E. coli rtPCR), were compared in terms of their sensitivity, ease of use, time to result and affordability. The three methods yielded a positive E. coli signal for 11.5, 10.8, and 11.5% of the 968 well water samples tested, respectively. One hundred and thirty-six (136) samples gave blue colonies on mFC agar and required confirmation. E. coli-specific rtPCR showed false-positive results in 23.5% (32/136) of cases. In terms of ease of use, Colilert was the simplest method to use while the MI method provided ease of use comparable to all membrane filtration methods. However, the mFC + E. coli rtPCR assay required highly trained employees for confirmation purposes. In terms of affordability, and considering contamination rate of well water samples tested, the Colilert method and the mFC + E. coli rtPCR assay were at least five times more costly than the MI agar method. Overall, compared with the other two methods tested, the MI agar method offers the most advantages to assess drinking water quality.

Molecular method for detection of total coliforms in drinking water samples.

This work demonstrates the ability of a bacterial concentration and recovery procedure combined with three different PCR assays targeting the lacZ, wecG, and 16S rRNA genes, respectively, to detect the presence of total coliforms in 100-ml samples of potable water (presence/absence test). PCR assays were first compared to the culture-based Colilert and MI agar methods to determine their ability to detect 147 coliform strains representing 76 species of Enterobacteriaceae encountered in fecal and environmental settings. Results showed that 86 (58.5%) and 109 (74.1%) strains yielded a positive signal with Colilert and MI agar methods, respectively, whereas the lacZ, wecG, and 16S rRNA PCR assays detected 133 (90.5%), 111 (75.5%), and 146 (99.3%) of the 147 total coliform strains tested. These assays were then assessed by testing 122 well water samples collected in the Québec City region of Canada. Results showed that 97 (79.5%) of the samples tested by culture-based methods and 95 (77.9%), 82 (67.2%), and 98 (80.3%) of samples tested using PCR-based methods contained total coliforms, respectively. Consequently, despite the high genetic variability of the total coliform group, this study demonstrated that it is possible to use molecular assays to detect total coliforms in potable water: the 16S rRNA molecular assay was shown to be as efficient as recommended culture-based methods. This assay might be used in combination with an Escherichia coli molecular assay to assess drinking water quality.

Detection of Escherichia coli colonies on confluent plates of chromogenic media used in membrane filtration.

MI agar (MI), Chromocult® Coliform agar ES (Chromocult), and DC with BCIG agar (DC) are chromogenic membrane filtration culture-based methods used to assess microbiological water quality. In this study, their ability to detect Escherichia coli colonies on confluent growth plates was determined by testing water samples containing increasing concentrations of a non-E. coli growing bacterium, Citrobacter youngae. Then, their ability to inhibit the growth of non-coliform bacteria was determined by simultaneously testing 603 well water samples. Results were compared with those obtained with mFC and Colilert® methods. Results showed that the E. coli count was difficult to determine on mFC, Chromocult and DC when non-E. coli colonies reached levels of 10(4)colony forming units (CFU)/100 mL. However, the E. coli count did not interfere with Colilert until non-E. coli colonies reached concentrations of 10(7)CFU/100 mL. No inhibition was observed with MI as E. coli colonies could be easily detected in the presence of at least 10(7)CFU/100 mL of C. youngae. Using well water samples, confluent growth plates were observed for 144, 177, and 185 of the 603 well water samples tested with the MI, Chromocult and DC methods, respectively. Among these confluent growth plates, E. coli colonies were not detected for 10, 20, and 31 water samples. However, they were detected by the mFC and/or Colilert methods. Thus, among the three methods tested, the MI method presented the lowest grow rate of atypic colonies and was the only one that presents no interference in the E. coli count.