Applying probability-weighted incubation period distributions to traditional wind rose methodology to improve public health investigations of Legionnaires' disease outbreaks.

In the event of a Legionnaires' disease outbreak, rapid location and control of the source of bacteria are crucial for outbreak management and regulation. In this paper, we describe an enhancement of the traditional wind rose for epidemiological use; shifting the focus of measurement from relative frequency of the winds speeds and directions to the relative volume of air carried, whilst also incorporating probability distributions of disease incubation periods to refine identification of the important wind directions during a cases window of exposure, i.e. from which direction contaminated aerosols most likely originated. The probability-weighted wind rose offers a potential improvement over the traditional wind rose by weighting the importance of wind measurements through incorporation of probability of exposure given an individual's time of symptom onset (obtained through knowledge of the incubation period), and by instead focusing on the volume of carrying air which offers better insight into the most probable direction of the source. This then provides a probabilistic distribution of which direction the wind was blowing around the time of infection. We discuss how the probability-weighted wind rose can be implemented during a Legionnaires' disease outbreak, and how outbreak control teams might use it as supportive evidence to identify the most likely direction of the contaminated source from the presumed site of exposure. In addition, this paper discusses how minor adjustments can be made to the method allowing the probability-weighted wind rose to be applied to other non-communicable airborne diseases, providing the disease's probability distribution for the incubation period distribution is well known.

Two Community Clusters of Legionnaires' Disease Directly Linked to a Biologic Wastewater Treatment Plant, the Netherlands.

A biologic wastewater treatment plant was identified as a common source for 2 consecutive Legionnaires' disease clusters in the Netherlands in 2016 and 2017. Sequence typing and transmission modeling indicated direct and long-distance transmission of Legionella, indicating this source type should also be investigated in sporadic Legionnaires' disease cases.

Legionnaires' Disease Outbreak on a Merchant Vessel, Indian Ocean, Australia, 2015.

Two cases of Legionnaires' disease and 1 of Pontiac fever occurred among the crew of a merchant ship operating off the shores of Australia. PCR assays identified potential sources in the ship's cabins. Modification of maritime regulations for Legionnaires' disease prevention in commercial vessels is needed for nonpassenger merchant ships.

Legionnaires' disease.

Since first identified in early 1977, bacteria of the genus Legionella are recognised as a common cause of community-acquired pneumonia and a rare cause of hospital-acquired pneumonia. Legionella bacteria multisystem manifestations mainly affect susceptible patients as a result of age, underlying debilitating conditions, or immunosuppression. Water is the major natural reservoir for Legionella, and the pathogen is found in many different natural and artificial aquatic environments such as cooling towers or water systems in buildings, including hospitals. The term given to the severe pneumonia and systemic infection caused by Legionella bacteria is Legionnaires' disease. Over time, the prevalence of legionellosis or Legionnaires' disease has risen, which might indicate a greater awareness and reporting of the disease. Advances in microbiology have led to a better understanding of the ecological niches and pathogenesis of the condition. Legionnaires' disease is not always suspected because of its non-specific symptoms, and the diagnostic tests routinely available do not offer the desired sensitivity. However, effective antibiotics are available. Disease notification systems provide the basis for initiating investigations and limiting the scale and recurrence of outbreaks. This report reviews our current understanding of this disease.

Legionella longbeachae detected in an industrial cooling tower linked to a legionellosis outbreak, New Zealand, 2015; possible waterborne transmission?

A legionellosis outbreak at an industrial site was investigated to identify and control the source. Cases were identified from disease notifications, workplace illness records, and from clinicians. Cases were interviewed for symptoms and risk factors and tested for legionellosis. Implicated environmental sources were sampled and tested for legionella. We identified six cases with Legionnaires' disease and seven with Pontiac fever; all had been exposed to aerosols from the cooling towers on the site. Nine cases had evidence of infection with either Legionella pneumophila serogroup (sg) 1 or Legionella longbeachae sg1; these organisms were also isolated from the cooling towers. There was 100% DNA sequence homology between cooling tower and clinical isolates of L. pneumophila sg1 using sequence-based typing analysis; no clinical L. longbeachae isolates were available to compare with environmental isolates. Routine monitoring of the towers prior to the outbreak failed to detect any legionella. Data from this outbreak indicate that L. pneumophila sg1 transmission occurred from the cooling towers; in addition, L. longbeachae transmission was suggested but remains unproven. L. longbeachae detection in cooling towers has not been previously reported in association with legionellosis outbreaks. Waterborne transmission should not be discounted in investigations for the source of L. longbeachae infection.

Legionnaires' disease in Europe, 2011 to 2015.

Under the coordination of the European Centre for Disease Prevention and Control (ECDC), the European Legionnaires' disease Surveillance Network (ELDSNet) conducts surveillance of Legionnaires' disease (LD) in Europe. Between 2011 and 2015, 29 countries reported 30,532 LD cases to ECDC (28,188 (92.3%) confirmed and 2,344 (7.7%) probable). Four countries (France, Germany, Italy and Spain) accounted for 70.3% of all reported cases, although their combined populations represented only 49.9% of the study population. The age-standardised rate of all cases increased from 0.97 cases/100,000 population in 2011 to 1.30 cases/100,000 population in 2015, corresponding to an annual average increase of 0.09 cases/100,000 population (95%CI 0.02-0.14; p = 0.02). Demographics and infection setting remained unchanged with ca 70% of cases being community-acquired and 80% occurring in people aged 50 years and older. Clinical outcome was known for 23,164 cases, of whom 2,161 (9.3%) died. The overall case fatality ratio decreased steadily from 10.5% in 2011 to 8.1% in 2015, probably reflecting improved reporting completeness. Five countries (Austria, Czech Republic, Germany, Italy, and Norway) had increasing age-standardised LD notification rates over the 2011-15 period, but there was no increase in notification rates in countries where the 2011 rate was below 0.5/100,000 population.

Heated birthing pools as a source of Legionnaires' disease.

In June 2014 Public Health England confirmed a case of Legionnaires' disease (LD) in a neonate following birth at home in a hired birthing pool incorporating a heater and a recirculation pump which had been filled in advance of labour. The case triggered a public health investigation and a microbiological survey of an additional ten heated birthing pools hired or recently hired to the general public across England. The birthing pool used by the parent of the confirmed case was identified as the source of the neonate's infection following detection of Legionella pneumophila ST48 in both patient and environmental samples. Legionella species were detected by quantitative polymerase chain reaction but not culture in a further three pools together with other opportunistic pathogens identified by culture and matrix-assisted laser desorption ionization-time of flight (MALDI-ToF) mass spectrometry. A Patient Safety Alert from NHS England and Public Health England was issued stating that heated birthing pools filled in advance of labour should not be used for home births. This recommendation remains in place. This investigation in conjunction with other recent reports has highlighted a lack of awareness regarding the microbiological safety of heated birthing pools and their potential to be a source of LD and other opportunistic infections. Furthermore, the investigation raised important considerations with regards to microbiological sampling and testing in such incidents. Public health authorities and clinicians should consider LD in the differential diagnosis of severe respiratory infection in neonates within 14 days of a water birth.

Hospital-acquired Legionella infections: an update on the procedures for controlling environmental contamination.

The waterborne healthcare-associated infections are mainly sustained by Legionella and Pseudomonas spp. Various water factors and plumbing characteristics, and the interaction with other water microorganisms are considered to be predictive of Legionella contamination. It is therefore mandatory to organize plans of surveillance, prevention and control in order to avoid disease appearance in immunosuppressed patients, with higher risk of death. Guidelines for the prevention of Legionnaires' disease have been published, benefiting those who face this problem, but definitive standardized solutions do not exist yet. Here we describe fifteen years of activity, during which our study group gathered interesting data on the control of Legionella contamination. Water disinfection is not generally sufficient to control the risk of infection, but a complex water safety plan should be developed, including system maintenance, training of staff and implementation of a clinical surveillance system aimed at early detection of cases. Concerning the control measures, we evaluated the effectiveness of different treatments suggested to reduce Legionella spp contamination, comparing our results with the current literature data. The performance ranking was highest for the filter, followed by boilers at high temperature, monochloramine and, at a lower level, chlorine dioxide; the effectiveness of hyperchlorination was limited, and thermal shock was even more ineffective.

Colonization of Legionella species in Turkish baths in hotels in Alanya, Turkey.

This study evaluated the prevalence of Legionella species in water samples collected from Turkish baths in hotels in Alanya, Turkey, from August 2003 to September 2013. Water samples were collected in 100-mL sterile containers and then concentrated by filtration. Heat treatment was used to eliminate other microorganisms from the samples, which were then spread on Legionella-selective-buffered charcoal yeast extract alpha (BCYE-α) agar and on BCYE-α agar supplemented with glycine, vancomycin, polymyxin, and cycloheximide. Cysteine-dependent colonies were identified by latex agglutination. In total, 135 samples from 52 hotels with Turkish baths were evaluated. Legionella species were identified in 11/52 (21.2%) hotels and 18/135 (13.3%) samples. The most frequently isolated species was Legionella pneumophila, with most isolates belonging to serogroups 6 (55.6%) and 1 (22.2%). The colony count was <100 colony-forming units (CFU) mL(-1) in nine samples, from 100 to 1000 CFU mL(-1) in six samples, and >1000 CFU mL(-1) in three samples. These findings suggest that the hot water systems of Turkish baths in hotels must be viewed as a possible source of travel-associated Legionnaires' disease, and preventative measures should be put in place.

Persistence of viable but nonculturable Legionella pneumophila state in hospital water systems: A hidden enemy?

There is little evidence of the long-term consequences of maintaining sanitary hot water at high temperatures on the persistence of Legionella in the plumbing system. The aims of this study were to describe the persistence and genotypic variability of L. pneumophila in a hospital building with two entirely independent hot water distribution systems, and to estimate the thermotolerance of the genotypic variants by studying the quantity of VBNC L. pneumophila. Eighty isolates from 55 water samples obtained between the years 2012-2017 were analyzed. All isolates correspond to L. pneumophila serogroup 6. The isolates were discriminated in four restriction patterns by pulsed-field gel electrophoresis. In one installation, pattern A + Aa predominated, accounting for 75.8 % of samples, while the other installation exhibited pattern B as the most frequent (81.8 % of samples; p < 0.001). The mean temperature of the isolates was: 52.6 °C (pattern A + Aa) and 55.0 °C (pattern B), being significantly different. Nine strains were selected as representative among patterns to study their thermotolerance by flow-cytometry after 24 h of thermic treatment. VBNC bacteria were detected in all samples. After thermic treatment at 50 °C, 52.0 % of bacteria had an intact membrane, and after 55 °C this percentage decreased to 23.1 %. Each pattern exhibited varying levels of thermotolerance. These findings indicate that the same hospital building can be colonized with different predominant types of Legionella if it has independent hot water installations. Maintaining a minimum temperature of 50 °C at distal points of the system would allow the survival of replicative L. pneumophila. However, the presence of Legionella in hospital water networks is underestimated if culture is considered as the standard method for Legionella detection, because VBNC do not grow on culture plates. This phenomenon can carry implications for the Legionella risk management plans in hospitals that adjust their control measures based on the microbiological surveillance of water.

Epidemiological investigation of a Legionnaires' disease outbreak in Christchurch, New Zealand: the value of spatial methods for practical public health.

Between April and August 2005 Christchurch, New Zealand experienced an outbreak of Legionnaires' disease. There were 19 laboratory-confirmed case including three deaths. Legionella pneumophila serogroup 1 (Lpsg1) was identified as the causative agent for all cases. A case-control study indicated a geographical association between the cases but no specific common exposures. Rapid spatial epidemiological investigation confirmed the association and identified seven spatially significant case clusters. The clusters were all sourced in the same area and exhibited a clear anisotropic process (noticeable direction) revealing a plume effect consistent with aerosol dispersion from a prevailing southwesterly wind. Four out of five cases tested had indistinguishable allele profiles that also matched environmental isolates from a water cooling tower within the centre of the clusters. This tower was considered the most probable source for these clusters. The conclusion would suggest a maximum dispersal distance in this outbreak of 11·6 km. This work illustrated the value of geostatistical techniques for infectious disease epidemiology and for providing timely information during outbreak investigations.

Assessing the risk of Legionnaires' disease: the inhalation exposure model and the estimated risk in residential bathrooms.

Legionella are widely found in the built environment. Patients with Legionnaires' disease have been increasing in Japan; however, health risks from Legionella bacteria in the environment are not appropriately assessed. We performed a quantitative health risk assessment modeled on residential bathrooms in the Adachi outbreak area and estimated risk levels. The estimated risks in the Adachi outbreak approximately corresponded to the risk levels exponentially extrapolated into lower levels on the basis of infection and mortality rates calculated from actual outbreaks, suggesting that the model of Legionnaires' disease in residential bathrooms was adequate to predict disease risk for the evaluated outbreaks. Based on this model, the infection and mortality risk levels per year in 10 CFU/100 ml (100 CFU/L) of the Japanese water quality guideline value were approximately 10(-2) and 10(-5), respectively. However, acceptable risk levels of infection and mortality from Legionnaires' disease should be adjusted to approximately 10(-4) and 10(-7), respectively, per year. Therefore, a reference value of 0.1 CFU/100 ml (1 CFU/L) as a water quality guideline for Legionella bacteria is recommended. This value is occasionally less than the actual detection limit. Legionella levels in water system should be maintained as low as reasonably achievable (<1 CFU/L).

Cluster of Legionnaires disease cases caused by Legionella longbeachae serogroup 1, Scotland, August to September 2013.

We report six confirmed cases of Legionnaires' disease in Scotland caused by Legionella longbeachae serogroup 1, identified over a four-week period in August­September 2013. All cases required admission to hospital intensive care facilities. All cases were amateur gardeners with frequent exposure to horticultural growing media throughout their incubation period. L. longbeachae was identified in five samples of growing media linked to five cases. Product tracing did not identify a common product or manufacturer.

Legionnaires disease in Europe, 2009-2010.

The surveillance of Legionnaires' disease (LD) in Europe is carried out by the European Legionnaires' Disease Surveillance Network (ELDSNet) and coordinated by the European Centre for Disease Prevention and Control (ECDC). All cases reported in 2009 and 2010 and meeting the European case definition were electronically transmitted to The European Surveillance System (TESSy) database. A total of 5,551 and 6,305 cases were reported by 29 European countries in 2009 and 2010, respectively. The age-standardised rate of all cases was 1.20 per 100,000 inhabitants in 2010, 12% higher than in 2009, which was consistent with the increasing trend observed since 2005. Most of this increase consisted of community-acquired cases reported by France, Germany and the Netherlands with dates of onset in August­September. The exceptionally hot summer of 2010 in some parts of Europe may have played a role in this increase.

Eight years of Legionnaires' disease transmission in travellers to a condominium complex in Las Vegas, Nevada.

Travel is a risk factor for Legionnaires' disease. In 2008, two cases were reported in condominium guests where we investigated a 2001 outbreak. We reinvestigated to identify additional cases and determine whether ongoing transmission resulted from persistent colonization of potable water. Exposures were assessed by matched case-control analyses (2001) and case-series interviews (2008). We sampled potable water and other water sources. Isolates were compared using sequence-based typing. From 2001 to 2008, 35 cases were identified. Confirmed cases reported after the cluster in 2001-2002 were initially considered sporadic, but retrospective case-finding identified five additional cases. Cases were more likely than controls to stay in tower 2 of the condominium [matched odds ratio (mOR) 6·1, 95% confidence interval (CI) 1·6-22·9]; transmission was associated with showering duration (mOR 23·0, 95% CI 1·4-384). We characterized a clinical isolate as sequence type 35 (ST35) and detected ST35 in samples of tower 2's potable water in 2001, 2002, and 2008. This prolonged outbreak illustrates the importance of striving for permanent Legionella eradication from potable water.

Direct healthcare costs of selected diseases primarily or partially transmitted by water.

Despite US sanitation advancements, millions of waterborne disease cases occur annually, although the precise burden of disease is not well quantified. Estimating the direct healthcare cost of specific infections would be useful in prioritizing waterborne disease prevention activities. Hospitalization and outpatient visit costs per case and total US hospitalization costs for ten waterborne diseases were calculated using large healthcare claims and hospital discharge databases. The five primarily waterborne diseases in this analysis (giardiasis, cryptosporidiosis, Legionnaires' disease, otitis externa, and non-tuberculous mycobacterial infection) were responsible for over 40 000 hospitalizations at a cost of $970 million per year, including at least $430 million in hospitalization costs for Medicaid and Medicare patients. An additional 50 000 hospitalizations for campylobacteriosis, salmonellosis, shigellosis, haemolytic uraemic syndrome, and toxoplasmosis cost $860 million annually ($390 million in payments for Medicaid and Medicare patients), a portion of which can be assumed to be due to waterborne transmission.

Development of a new CARD-FISH protocol for quantification of Legionella pneumophila and its application in two hospital cooling towers.

AIMS: Open cooling towers are frequent sources of infections with Legionella pneumophila. The gold standard for the detection of Leg. pneumophila is based on cultivation lasting up to 10 days and detecting only culturable cells. Alternative fluorescence in situ hybridization (FISH) protocols have been proposed, but they result in faint fluorescence signals and lack specificity because of cross-hybridization with other Legionella species. Our aim was thus to develop a new FISH protocol for rapid and specific detection of Leg. pneumophila in water samples. METHODS AND RESULTS: A novel catalysed reporter deposition FISH (CARD-FISH) protocol for the detection of Leg. pneumophila was developed, which significantly enhanced signal intensity as well as specificity of the probe through the use of a novel competitor probe. The developed protocol was compared with the culture method for monitoring the seasonal development of culturable and nonculturable Leg. pneumophila in two hospital cooling tower systems. Seasonal fluctuations of Leg. pneumophila concentrations detected via CARD-FISH were related to the development of the total bacterial community in both cooling towers, with temperature and biocide as the main factors controlling this development. CONCLUSIONS: Our results clearly showed that the majority of the Leg. pneumophila cells were in a nonculturable state. Thus, detection of Leg. pneumophila with culture methods may underestimate the total numbers of Leg. pneumophila present. SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid, sensitive and specific detection and quantification of Leg. pneumophila in water systems is prerequisite for reliable risk estimation. The new protocol significantly improves current methodology and can be used to monitor and screen for Leg. pneumophila concentrations in cooling towers or other water systems.

The application of geographic information systems and spatial data during Legionnaires disease outbreak responses.

A literature review was conducted to highlight the application and potential benefit of using geographic information systems (GIS) during Legionnaires' disease outbreak investigations. Relatively few published sources were identified, however, certain types of data were found to be important in facilitating the use of GIS, namely: patient data, locations of potential sources (e.g. cooling towers), demographic data relating to the local population and meteorological data. These data were then analysed to gain a better understanding of the spatial relationships between cases and their environment, the cases' proximity to potential outbreak sources, and the modelled dispersion of contaminated aerosols. The use of GIS in an outbreak is not a replacement for traditional outbreak investigation techniques, but it can be a valuable supplement to a response.

Preliminary report: outbreak of Legionnaires disease in a hotel in Calp, Spain, update on 22 February 2012.

Research is ongoing on eighteen cases of Legionellosis, including four deaths, identified among tourists and employees in a hotel in Calp, Spain. Cases occurred during a period of two months, indicating the possibility of a point-source transmission at the hotel. An environmental investigation identified several positive samples in the hotel, which as a precautionary measure, was closed until requested improvements were made. Surveillance measures currently remain active.