Legionella spp. Colonization in Water Systems of Hotels Linked with Travel-Associated Legionnaires’ Disease
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sample Collection
2.2. Risk Assessment, Data Collection and Corrective Action
2.3. Plate Culture Method
2.4. Typing of Legionella Isolates
2.5. Identification—MALDI-TOF Mass Spectrometry
2.6. Statistical Analysis
3. Results
3.1. Descriptive Data
3.2. Isolation and Identification of Legionella Species
3.3. Univariate Examination of Factors
3.4. Inspection Results and Implementation of WSPs
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sample Description | No of Positive Samples/Total (%) | p-Values (Proportional z-Test) | ||||||
---|---|---|---|---|---|---|---|---|
Legionella spp. | L. pneumophila Serogroup 1 | L. pneumophila Serogroup 2–15 | L. non-pneumophila | Serogroup 1 vs. Serogroup 2–15 | Serogroup 1 vs. non-pneumophila | Serogroup 2–15 vs. non-pneumophila | ||
Cold water system | Cold water first catch sample (rooms closest to boilers) | 31/127 (24.41) | 6/127 (4.72) | 16/127 (12.60) | 17/127 (13.39) | 0.026 * | 0.016 * | 0.852 |
Cold water first catch sample (rooms distal to boilers) | 102/212 (48.11) | 27/212 (12.74) | 61/212 (28.77) | 32/212 (15.09) | <0.001 ** | 0.468 | <0.001 ** | |
Cold water sample after two min flush (rooms closest to boilers) | 21/87 (24.14) | 6/87 (6.90) | 9/87 (10.34) | 11/87 (12.64) | 0.419 | 0.202 | 0.634 | |
Cold water sample after two min flush (rooms distal to boilers) | 25/106 (23.58) | 6/106 (5.66) | 16/106 (15.09) | 8/106 (7.55) | 0.024 * | 0.580 | 0.083 | |
Hotel room cold water first catch sample | 4/8 (50.00) | 1/8 (12.50) | 2/8 (25.00) | 1/8 (12.50) | 0.522 | 1.000 | 0.552 | |
Hotel room cold water sample after two minutes flush | 1/7 (14.29) | 1/7 (14.29) | - | - | - | - | - | |
Hot water system | Hotel room hot water first catch sample | 5/8 (62.50) | 1/8 (12.50) | 4/8 (50.00) | 1/8 (12.50) | 0.106 | 1.000 | 0.106 |
Hotel room hot water sample after two minutes flush | 3/7 (42.86) | 1/7 (14.29) | 3/7 (42.86) | - | 0.237 | - | - | |
Returning hot water sample | 26/76 (34.21) | 7/76 (9.21) | 18/76 (23.68) | 6/76 (7.89) | 0.016 * | 0.771 | 0.008 ** | |
Hot water first catch sample (rooms distal to boilers) | 45/142 (31.69) | 14/142 (9.86) | 27/142 (19.01) | 16/142 (11.27) | 0.028 * | 0.966 | 0.069 | |
Hot water sample after two min flush (rooms closest to boilers) | 47/110 (42.73) | 22/110 (20.00) | 28/110 (25.45) | 12/110 (10.91) | 0.335 | 0.062 | 0.005 | |
Hot water sample after two min flush (rooms distal to boilers) | 81/158 (51.27) | 32/158 (20.25) | 60/158 (37.97) | 19/158 (12.03) | <0.001 ** | 0.047* | <0.001 * | |
Hot water sample heated from solar panels | 2/7 (28.57) | 1/7 (14.29) | 1/7 (14.29) | - | 1.000 | - | - | |
Water sample directly from boiler | 38/115 (33.04) | 9/115 (7.83) | 27/115 (23.48) | 8/115 (6.96) | <0.001 ** | 0.801 | <0.001 ** | |
Water sample from alternative heating source | 1/3 (33.33) | 1/3 (33.33) | 1/3 (33.33) | - | 1.000 | - | - | |
Water sample from spa establishment showers | 3/11 (27.27) | 1/11 (9.09) | 2/11 (18.18) | - | 0.534 | - | - | |
Water sample from swimming pool showers | 32/182 (17.58) | 7/182 (3.85) | 20/182 (10.99) | 10/182 (5.49) | 0.009 ** | 0.548 | 0.056 | |
Hot water first catch sample (rooms closest to boilers) | 194 (51.55) | 38/194 (19.59) | 59/194 (30.41) | 27/194 (13.92) | 0.014 * | 0.139 | <0.001 ** |
Sample Type | Number of Samples | |||
---|---|---|---|---|
Low (≤1.000) | Medium (>1.000 & <10.000) | High (≥10.000) | Total | |
Cold water distribution system | 177 (57.84) | 79 (35.11) | 64 (41.56) | 320 |
Hot water distribution system | 129 (42.16) | 144 (64.00) | 87 (56.49) | 360 |
Sediment from room shower water sample and from filtering systems | 0 (0.00) | 2 (0.89) | 3 (1.95) | 5 |
Soil | 0 (0.00) | 0 (0.00) | 0 (0.00) | 0 |
Total (n) | 306 (44.67) | 225 (32.80) | 154 (22.48) | 685 |
Legionella Species and Serogroup (sg) | Legionella spp. Concentration (CFU/L) | |||
---|---|---|---|---|
Species | ≤103 (%) | >103 and <104 (%) | ≥104 (%) | Total (%) |
L. sg1 | 51 (68.00) | 19 (25.33) | 5 (6.67) | 75 (22.20) |
L. sgs 2-15 | 99 (54.10) | 45 (24.59) | 39 (21.31) | 183 (49.39) |
L. non-pneumophila | 63 (55.75) | 28 (24.78) | 22 (19.47) | 113 (30.46) |
Total number of samples | 222 (59.84) | 84 (22.64) | 72 (19.41) | 320 (16.98) |
Parameter | Number of Samples | Positive Samples (≥50 CFU/L) Legionella Species and Serogroup (sg) | ||||
---|---|---|---|---|---|---|
Legionella spp. * | sg 1 | sg 2–15 | L. non-pneumophila | |||
Hot water temperature (Celsius) | 20–40 | 197 | 62 | 19 | 45 | 16 |
41–50 | 285 | 90 | 40 | 51 | 25 | |
51–55 | 170 | 38 | 11 | 26 | 10 | |
>55 | 304 | 17 | 5 | 13 | 3 | |
Total | 956 | 207 | 75 | 135 | 54 | |
Cold water temperature (Celsius) | 10–20 | 140 | 20 | 6 | 12 | 2 |
21–25 | 497 | 64 | 16 | 36 | 21 | |
26–30 | 315 | 65 | 12 | 33 | 34 | |
>30 | 71 | 15 | 2 | 8 | 8 | |
Total | 1023 | 164 | 36 | 89 | 65 | |
Residual chlorine (mg/L) | 0–0.20 | 424 | 110 | 29 | 48 | 50 |
0.21–0.50 | 226 | 31 | 5 | 16 | 12 | |
>0.51 | 345 | 20 | 2 | 12 | 8 | |
Total | 995 | 161 | 36 | 76 | 70 |
Risk Factors | Odds Ratio (95% Confidence Interval) |
---|---|
Boiler outflowing water temperature <60 °C | 27.5455 (1.6349–464.1095) |
Boiler returning hot water temperature <55 °C | 9.1698 (1.1613–72.4041) |
No use of alternative disinfection procedures | 7.2528 (2.2864–23.0074) |
Hot water temperature <55 °C | 5.9124 (3.8358–9.4106) |
Boiler returning hot water temperature <50 °C | 4.6667 (1.5273–14.2593) |
Incorrect application of WSP measures | 3.4593 (2.0965–5.7078) |
Residual Chlorine <0.2 mg/L | 3.3242 (2.3876–4.6595) |
Start of season | 2.2562 (1.3326–3.8200) |
Star classification <4 | 1.982 (1.6442–2.3894) |
Exclusive use of solar panels and hot water temperature <55 °C | 1.9438 (1.0398–3.6335) |
Absence of a Water Safety Plan | 1.7459 (1.4109–2.1604) |
Population using municipality water distribution system <10.000 residents | 1.4624 (1.2243–1.7469) |
Cold water temperature >25 °C | 1.4238 (1.0979–1.8464) |
Residual Chlorine <0.2 mg/L & pH out of range | 1.4075 (0.6882–2.8786) |
No guidance by the public health authority (1st inspection) | 1.249 (1.0444–1.4965) |
End of season | 1.0643 (0.7742–1.4630) |
High season period | 1.0578 (0.8626–1.2971) |
Non-automated disinfection system | 0.9769 (0.6039–1.5802) |
Exclusive use of solar panels | 0.9396 (0.5886–1.5000) |
Seasonal operation | 0.8499 (0.6968–1.0367) |
Hot water distribution system—distal room to boiler | 0.8217 (0.6022–1.1212) |
Unsatisfactory operations according to the checklist | 0.6672 (0.3539–1.2576) |
pH out of range | 0.5481 (0.3432–0.8753) |
Number of rooms >80 | 0.4427 (0.3652–0.5366) |
Number of beds >200 | 0.4351 (0.3529–0.5366) |
Groundwater as a source of water supply | 0.3717 (0.2564–0.5388) |
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Papadakis, A.; Keramarou, M.; Chochlakis, D.; Sandalakis, V.; Mouchtouri, V.A.; Psaroulaki, A. Legionella spp. Colonization in Water Systems of Hotels Linked with Travel-Associated Legionnaires’ Disease. Water 2021, 13, 2243. https://doi.org/10.3390/w13162243
Papadakis A, Keramarou M, Chochlakis D, Sandalakis V, Mouchtouri VA, Psaroulaki A. Legionella spp. Colonization in Water Systems of Hotels Linked with Travel-Associated Legionnaires’ Disease. Water. 2021; 13(16):2243. https://doi.org/10.3390/w13162243
Chicago/Turabian StylePapadakis, Antonios, Maria Keramarou, Dimosthenis Chochlakis, Vassilios Sandalakis, Varvara A. Mouchtouri, and Anna Psaroulaki. 2021. "Legionella spp. Colonization in Water Systems of Hotels Linked with Travel-Associated Legionnaires’ Disease" Water 13, no. 16: 2243. https://doi.org/10.3390/w13162243