Erica Leoni

Legionella waterline colonization: detection of Legionella species in domestic, hotel and hospital hot water systems

Aims:  An evaluation was made of the prevalence of Legionella species in hot water distribution systems in the city of Bologna (Italy) and their possible association with bacterial contamination (total counts and Pseudomonadaceae) and the chemical characteristics of the water (pH, Ca, Mg, Fe, Mn, Cu, Zn and Total Organic Carbon, TOC).

Legionella Infection Risk from Domestic Hot Water

We investigated Legionella and Pseudomonas contamination of hot water in a cross-sectional multicentric survey in Italy. Chemical parameters (hardness, free chlorine, and trace elements) were determined. Legionella spp. were detected in 33 (22.6%) and Pseudomonas spp. in 56 (38.4%) of 146 samples. Some factors associated with Legionella contamination were heater type, tank distance and capacity, water plant age, and mineral content. Pseudomonas presence was influenced by water source, hardness, free chlorine, and temperature. Legionella contamination was associated with a centralized heater, distance from the heater point >10 m, and a water plant >10 years old. Furthermore, zinc levels of <20 μg/L and copper levels of >50 μg/L appeared to be protective against Legionella colonization. Legionella species and serogroups were differently distributed according to heater type, water temperature, and free chlorine, suggesting that Legionella strains may have a different sensibility and resistance to environmental factors and different ecologic niches.

Comparison of selective procedures for isolation and enumeration of Legionella species from hot water systems

E. LEONI AND P.P. LEGNANI. 2001. Various sample pre‐treatment techniques and different growth media for the isolation of Legionellae from hot water supplies in public buildings were compared. A total of 102 hot water samples from taps and showers was examined. The highest recovery frequency was obtained with the heat pre‐treatment method and using the selective medium GVPC. However, the results differed according to the concentration of legionellas. In the case of low plate counts (≤5000 cfu l−1), the heat pre‐treatment technique gave a significantly higher percentage of positive samples compared with other techniques (P < 0·05). With increasing concentration, the differences between the procedures decreased until they became statistically not significant for concentrations above 50 000 cfu l−1. The direct inoculum method allowed a significantly higher detection of concentrations (P < 0·001) compared with heat and acid decontamination methods, which brought about a 67–68% reduction in detectable Legionellae. Heat decontamination techniques show greater sensitivity and specificity. However, they underestimate the number of legionellas. In environmental surveillance programmes, this underestimate must be taken into consideration when assessing the health risk.

Prevalence of legionella spp. In swimming pool environment

A study was carried out to evaluate the prevalence of bacteria of the Legionella genus in the water from the pools and showers of 12 swimming pools in the city of Bologna (Italy). Sampling took place four times for every establishment, once for each season of the year. Legionella spp. were found in 2 of the 48 pool water samples (L. micdadei and L. bozenanii) and in 27 of the 48 samples taken from the hot water of the showers: 19 were positive for L. pneumophila (10-19,250 cfu/l) and 18 for other species (20-6000 cfu/l). The contamination was seen to be consistent throughout the year, since the same species of legionellae were isolated at different samplings and in concentrations that did not reveal any seasonal trend. The legionellae and the Gram negative bacteria were not found in shower water with a temperature above 43 degrees C. However, in the samples of shower water with a lower temperature a statistically significant inverse correlation was seen between legionellae and Pseudomonas aeruginosa (r = -0.51; p<0.01) as well as between legionellae and Gram negative bacteria (r = -0.70; p<0.01). The potential risk of contracting infections from Legionella spp. in the swimming pool environment does not seem to be linked to the pool water, but to that of the showers. The water temperature of showers should therefore be maintained at a level high enough to prevent the reproduction of these bacteria.

Prevalence of mycobacteria in a swimming pool environment

A study was performed to evaluate the prevalence of non‐tubercular mycobacteria in swimming pool environments. The bacteria in question were found in 88·2% of pool water samples. The most frequent species were Mycobacterium gordonae (73·5% of samples; range 1–840 cfu 100 ml−1), M. chelonei (38·2% 2–360 cfu 100 ml−1) and M. fortuitum (35·3% 2–250 cfu 100 ml−1). The same species were also recovered from the water at the different phases of the treatment cycle, with relative percentages similar to those of the pool water. Shower floors and pool edges also presented high concentrations of the mycobacteria (100% of samples) and M. marinum was isolated from the surfaces of pool edges on two occasions (4·5% of samples). The swimming pool environment provides a suitable habitat for the survival and reproduction of mycobacteria. Although mycobacteria are common in swimming pools, human mycobacterial disease associated with their use is rare. Apart from superficial infections with M. marinum, the risk of more serious diseases in subjects with weakened immune systems should not be underestimated, given the widespread presence of mycobacteria that are possible opportunistic pathogens and the direct contact bathers have with the water and aerosol.

Distribution of indicator bacteria and bacteriophages in shellfish and shellfish‐growing waters

Shellfish (mussels and clams) and shellfish‐growing waters were examined for indicator bacteria according to the EC regulations, Salmonella spp., coliphages and anti‐Salmonella phages. Samples were collected both from natural‐growing areas along the coast and from authorized shellfish‐harvesting beds. The coastal area was affected by organic pollution and extensive faecal contamination and, according to the legal requirements, was unsuitable for shellfish farming. The shellfish collected along the coast also showed faecal contamination at levels which did not conform to legal standards. No significant differences were observed between the frequency of isolation of somatic coliphages and indicator bacteria from sea water. In contrast, both the authorized and wild coastal shellfish were contaminated by coliphages at a significantly higher level than the corresponding bacterial indicators for faecal contamination (χ2 test, P < 0·01). Coliphage concentrations were significantly correlated with faecal indicators in marine waters (P < 0·001) and sediments (P < 0·05), but no correlation was found in shellfish, thus showing their low specificity as indicators of faecal pollution of human origin in shellfish of economic importance.

Control of Legionella pneumophila contamination in a respiratory hydrotherapy system with sulfurous spa water.

OBJECTIVE To evaluate the effectiveness of different disinfection treatments in a spa water system contaminated by Legionella pneumophila and associated with a case of Legionella pneumonia. DESIGN During an 18-month period, the spa water was analyzed by taking samples from the well, the recirculation line, and the final distribution devices (nebulizers and nasal irrigators). Various attempts were made to eradicate Legionella organisms by chemical and thermal shock. The final protocol consisted of heat shock treatment at 70 degrees C-75 degrees C for 3 hours, 2 nights per week, followed by a lowering of the water temperature to 30 deg C+/-1 deg C for use in the plant. In addition, 3 times a week superheated steam (at a pressure of 1 atmosphere) was introduced for 1 hour into the nebulization machines. SETTING A spa at which sulfurous water was used for hydrotherapy by means of aerosol and nasal irrigation. PATIENT A 74-year-old woman with legionnaires disease. RESULTS After the case of infection occurred, L. pneumophila was isolated from the recirculation line at a concentration of 400,000 cfu/L and from the nebulizers and nasal irrigators at levels ranging from 3,300 to 1,800,000 cfu/L. The colonizing organisms consisted of a mixture of L. pneumophila serogroup 1 (12%) and serogroup 5 (88%). The shock treatment with chlorine dioxide and peracetic acid resulted in the eradication of Legionella organisms from the recirculation line but not from the water generated from the final distribution devices. After the restructuring of the plant and the application of thermal shock protocol, an evaluation after 12 months revealed no evidence of Legionella contamination. CONCLUSION To prevent Legionella colonization, disinfection treatment is effective if associated with carefully selected materials, good circuit design, and good maintenance practices.

An outbreak of Salmonella Hadar associated with roast rabbit in a restaurant

In August 1997, an outbreak of gastroenteritis from SalmonellaHadar phage type 2 occurred among customers of a restaurant in Rimini (Emilia-Romagna region, Italy). Twenty-nine people who had eaten food prepared in the restaurant on 2 or 3 August had symptoms of acute gastroenteritis. The infection was culture-confirmed in 24 cases and the stool specimens of four healthy people were positive for SalmonellaHadar. Twelve people had to be hospitalized and a 3-year old girl died. The case–control study identified roast rabbit as the likely vehicle of infection (OR: 6.00; CI 95%: 1.65–22.83). The microbiological investigation carried out on food taken from the restaurant confirmed high levels of SalmonellaHadar in a sample of roast rabbit. Since the rabbit was well cooked, the food contamination likely occurred after cooking. Poor hygienic conditions found in the restaurant, together with inappropriate food-handling practices and inadequate storage temperatures may have contributed to spread to other foods and the severity of the outbreak.

Effect of different disinfection protocols on microbial and biofilm contamination of dental unit waterlines in community dental practices.

Output water from dental unit waterlines (DUWLs) may be a potential source of infection for both dental healthcare staff and patients. This study compared the efficacy of different disinfection methods with regard to the water quality and the presence of biofilm in DUWLs. Five dental units operating in a public dental health care setting were selected. The control dental unit had no disinfection system; two were disinfected intermittently with peracetic acid/hydrogen peroxide 0.26% and two underwent continuous disinfection with hydrogen peroxide/silver ions (0.02%) and stabilized chlorine dioxide (0.22%), respectively. After three months of applying the disinfection protocols, continuous disinfection systems were more effective than intermittent systems in reducing the microbial contamination of the water, allowing compliance with the CDC guidelines and the European Council regulatory thresholds for drinking water. P. aeruginosa, Legionella spp, sulphite-reducing Clostridium spores, S. aureus and β-haemolytic streptococci were also absent from units treated with continuous disinfection. The biofilm covering the DUWLs was more extensive, thicker and more friable in the intermittent disinfection dental units than in those with continuous disinfection. Overall, the findings showed that the products used for continuous disinfection of dental unit waterlines showed statistically better results than the intermittent treatment products under the study conditions.

Impact of a risk management plan on Legionella contamination of dental unit water.

The study aimed to assess the prevalence of Legionella spp. in dental unit waterlines of a dental clinic and to verify whether the microbiological parameters used as indicators of water quality were correlated with Legionella contamination. A risk management plan was subsequently implemented in the dental health care setting, in order to verify whether the adopted disinfection protocols were effective in preventing Legionella colonization. The water delivered from syringes and turbines of 63 dental units operating in a dental clinic, was monitored for counts of the heterotrophic bacteria P. aeruginosa and Legionella spp. (22 °C and 37 °C). At baseline, output water from dental units continuously treated with disinfection products was more compliant with the recommended standards than untreated and periodically treated water. However, continuous disinfection was still not able to prevent contamination by Legionella and P. aeruginosa. Legionella was isolated from 36.4%, 24.3% and 53.3% of samples from untreated, periodically and continuously treated waterlines, respectively. The standard microbiological parameters used as indicators of water quality proved to be unreliable as predictors of the presence of Legionella, whose source was identified as the tap water used to supply the dental units. The adoption of control measures, including the use of deionized water in supplying the dental unit waterlines and the application of a combined protocol of continuous and periodic disinfection, with different active products for the different devices, resulted in good control of Legionella contamination. The efficacy of the measures adopted was mainly linked to the strict adherence to the planned protocols, which placed particular stress on staff training and ongoing environmental monitoring.