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Legionella-related disease is caused by an intracellular bacteria mainly living in water. Contamination results from inhalation of Legionella sp containing aerosolized water. Main risk factors are tobacco, immunodeficiency, and advanced age. Antigenuria is the cornerstone of the diagnosis. Immunocompromised patients, more commonly infected with non pneumophilaLegionella, present negative antigenuria, and culture and PCR are essential for the diagnosis. Legionnaires' disease may be severe, especially in elderly and/or immunocompromised patients. Mortality rate varies from 10 % in the general population to 50 % in intensive care. Treatment is based on macrolides or fluoroquinolones. Antibiotic resistance is very rare., (Copyright © 2019. Published by Elsevier Masson SAS.)

Objectives: Infections caused by Legionella are the leading cause of waterborne disease outbreaks in the United States. We investigated a large outbreak of Legionnaires' disease in New York City in summer 2015 to characterize patients, risk factors for mortality, and environmental exposures., Methods: We defined cases as patients with pneumonia and laboratory evidence of Legionella infection from July 2 through August 3, 2015, and with a history of residing in or visiting 1 of several South Bronx neighborhoods of New York City. We describe the epidemiologic, environmental, and laboratory investigation that identified the source of the outbreak., Results: We identified 138 patients with outbreak-related Legionnaires' disease, 16 of whom died. The median age of patients was 55. A total of 107 patients had a chronic health condition, including 43 with diabetes, 40 with alcoholism, and 24 with HIV infection. We tested 55 cooling towers for Legionella, and 2 had a strain indistinguishable by pulsed-field gel electrophoresis from 26 patient isolates. Whole-genome sequencing and epidemiologic evidence implicated 1 cooling tower as the source of the outbreak., Conclusions: A large outbreak of Legionnaires' disease caused by a cooling tower occurred in a medically vulnerable community. The outbreak prompted enactment of a new city law on the operation and maintenance of cooling towers. Ongoing surveillance and evaluation of cooling tower process controls will determine if the new law reduces the incidence of Legionnaires' disease in New York City.

Legionella spp. are pathogens that can cause Legionnaires' disease in humans through inhalation of contaminated aerosols. The principal reservoir for these microorganisms is water, but Legionella spp. have been isolated from composted vegetable and plant material, and from many potting mixes as well. In Australia, there have been several cases of Legionnaires' disease in which Legionella longbeachae has been isolated from potting soils. In Switzerland, the source of infection cannot always be identified as water or cooling towers: therefore, we have investigated 46 commercially available potting soils in Switzerland to determine the presence of Legionella spp. We were able to detect Legionella spp. in 45.7% (21/46) of the potting soil samples analysed by culture. Legionella pneumophila was present in 19.6% (9/46) of the samples and L. pneumophila serogroup 1 in 6.5% (3/46). Quantification by both culture and quantitative real-time PCR revealed high concentrations of legionellae in potting soils, ranging between 10(3) CFU/g and 10(5) CFU/g and 10(4) genomic units (GU)/g and 10(6) GU/g, respectively. Thus, potting soils may represent an alternative reservoir for Legionella spp. in Switzerland.

The authors discuss the case of a 79-year-old patient who suffered from a swallowing disorder and developed Legionnaires' disease 2 days after her dismissal from an orthopedics ward, where she had recovered from hip surgery. To determine the source of the Legionnaires' disease, the authors performed an environmental investigation, which included a national, standardized questionnaire and a microbiological investigation of suspected sources. The investigation revealed ice from an ice-making machine in the hospital as the most probable source of the infection through aspiration, even though the hospital had rigorously adhered to strict assessment and decontamination schedules. The infectious serogroup was one that was not common to the area. From the data available, the authors inferred that a dose of 1-2000 colony-forming units might have caused Legionnaires' disease in this patient.

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 <100 microg/L and copper levels of >50 microg/L appeared to be protective against Legionella colonization [corrected]. 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.

There have been reported outbreaks of Legionnaires' disease at hospitals and industrial facilities, which prompted the development of various preventive measures. For example, Ford has been developing and implementing such a measure at its facilities worldwide to provide technical guidance for controlling Legionella in water systems. One of the key issues for implementing the measure is the selection of a disinfectant(s) and optimum conditions for its use. Therefore, available publications on various disinfectants and disinfection processes used for the inactivation of Legionella bacteria were reviewed. Two disinfection methods were reviewed: chemical and thermal. For chemical methods, disinfectants used were metal ions (copper and silver), oxidizing agents (halogen containing compounds [chlorine, bromine, iodine, chlorine dioxide, chloramines, and halogenated hydantoins], ozone, and hydrogen peroxide), non-oxidizing agents (heterocyclic ketones, guanidines, thiocarbamates, aldehydes, amines, thiocyanates, organo-tin compounds, halogenated amides, and halogenated glycols), and UV light. In general, oxidizing disinfectants were found to be more effective than non-oxidizing ones. Among oxidizing agents, chlorine is known to be effective and widely used. Among non-oxidizing agents, 2,2-dibromo-3-nitropropionamide appears to be the most effective followed by glutaraldehyde. Isothiazolin (known as Kathon), polyhexamethylene biguanide, and 2-bromo-2-nitropropionamide (known as Bronopol) were found to be less effective than glutaraldehyde. Thermal disinfection is effective at > 60 degrees C (140 degrees F).

Since Legionnaires' Disease (LD) was first reported in 1976, outbreaks have been associated with airborne transmission of Legionella bacteria through cooling towers, showers, and other aerosolizing devices (1). However, most LD cases are sporadic, and the source and mode of infection in many cases are unknown. Infections with one species, Legionella longbeachae, have been associated with gardening and use of potting soil in Australia and Japan (2,3). This report summarizes the findings of LD investigations in California, Oregon, and Washington, that suggest that transmission from potting soil has occurred for the first time in the United States, and that active surveillance and case finding are warranted to explore this association.

A lack of standardization of environmental monitoring techniques for Legionella spp. complicates the interpretation of results and comparisons of results from different institutions. A comparative assessment of techniques recommended by the Centers for Disease Control and Prevention, the Hygiene Institute (Graz, Austria), and our laboratory was performed. Variables investigated were sampling method (swabbing and collection of water samples [250 ml] before and after swabbing), method of concentration (none, filtration, and centrifugation), acid buffer treatment (no acid treatment, treatment for 3 min, and treatment for 15 min), and choice of medium (five formulations of buffered charcoal yeast extract agar with glycine, vancomycin, polymyxin B, anisomycin, or cycloheximide). Thirty-three sites in seven hospital buildings were studied. Recovery by swab correlated with recovery from water after swabbing (P < 0.05). However, the quantity of Legionella spp. recovered from swab specimens (mean, 3.0 x 10(4) CFU per swab) was greater than that recovered from water (mean, 4.7 x 10(3) CFU/250 ml). Filtration resulted in recovery rates (mean, 5.2 x 10(3) CFU/250 ml) higher than those by centrifugation (mean, 2.3 x 10(3) CFU/250 ml). Three minutes of acid buffer treatment to reduce overgrowth by commensal flora did not improve selectivity or sensitivity for Legionella spp. if glycine-containing selective media were used. Fifteen minutes of acid buffer treatment reduced recovery compared with that after a 3-min treatment. All glycine-containing media tested effectively inhibited background flora, but one selective medium containing dyes, glycine, vancomycin, and polymyxin B (DGVP) resulted in the greatest quantitative recovery of Legionella pneumophila. Use of buffered charcoal yeast extract agar and the acid buffer treatment gave the greatest recovery of non-pneumophila species. A standardized protocol with an emphasis on the culturing of swab samples is presented.

We studied all cases of nosocomial pneumonia at our 800-bed tertiary care hospital from September 1983 to September 1987. Of the 813 cases of nosocomial pneumonia, 31 (3.8%) were definite (isolation of organism or fourfold rise in titer) and 21 (2.5%) were possible cases (single or stable antibody titer of greater than or equal to 1:256) of legionnaires' disease. The definite cases involved a more severe form of pneumonia and a significantly higher mortality rate--64% versus 14% (p less than 0.0009) compared with the possible cases. Despite attempted comprehensive surveillance, only four (13%) of the definite cases of legionnaires' disease were found that would not have been diagnosed if the study were not ongoing. The yield from adequate (4- to 6-week convalescent serum samples) serologic testing was 5%, whereas the yield from sputum culture was 11%. We conclude that targeted surveillance of immunosuppressed patients with nosocomial pneumonia by culture of respiratory tract secretions for Legionella pneumophila is adequate for monitoring for the presence of legionnaires' disease in a hospital.

Guidelines for the prevention of nosocomial pneumonia specify that only sterile fluids should be used for aerosol therapy; however, this recommendation may not be uniformly followed. Thirteen patients with nosocomial pneumonia due to Legionella pneumophila serogroup 3 (Lp3) were identified at a community hospital in the period from 1984 through 1988; 12 patients (92%) had chronic obstructive pulmonary disease; and 9 patients (69%) died. An epidemiologic investigation suggested that the use of nebulizers to deliver medication was associated with acquiring legionnaires' disease. The hospital potable water system was contaminated with Lp3, and a survey indicated that tap water was commonly used to wash medication nebulizers. Lp3 in respirable-size droplets was isolated from aerosols generated by a nebulizer containing Lp3 at one-tenth the concentration found in the hospital potable water. These findings support the recommendation that only sterile fluids be used for filling or cleaning respiratory care equipment and suggest that this guideline is not universally followed.

The study of the biological properties of monoclonal antibodies (McAb) to L. pneumophila cytolysin has been carried out. These McAb have been shown to possess no capacity for the in vitro neutralization of cytolysin and the protection of guinea pigs from aerosol infection with L. pneumophila. Still the protective effect of the McAb under study has been observed in experiments with the intraperitoneal infection of guinea pigs, which is indicative of the possibility, in principle, of involving humoral immunity into the protection of the body from Legionella infection.

Legionellae are opportunistic facultative intracellular pathogens with several antigens and toxic components which may function synergistically in contribution to pathogenesis and immunity. The serogroup specific antigens are associated with endotoxic activities of the bacteria and are potent adjuvants in inducing hypersensitivity and potentiating antibody responses to various antigens. Two protein toxins interfere with oxidative metabolism of host cells and are potential inhibitors of intracellular bactericidal activities. Tissue damage in the host appears to be caused by lysis of infected cells with release of toxic substances of host and bacterial origins. In host defense, the serogroup antigens provide primarily group-specific immunity, and the acid soluble protein toxin induces cross-protection among different serogroups. The combined effects of antibodies to these antigens in promoting phagocytosis and abrogating toxicity would enhance intracellular killing of the invading bacteria. The serogroup antigen complexes may also function as a potent adjuvant to modulate host responses to the invading bacteria.

To investigate the pathogenesis of Legionnaires disease at a molecular level, we mutated by directed allelic exchange a gene encoding a Legionella pneumophila-specific 24,000-dalton (Da) surface protein. Southern hybridization and immunoblot analyses demonstrated that the predicted DNA rearrangement occurred in L. pneumophila with a specific loss of 24-kDa antigen expression. Compared with its isogenic parent, the mutant was significantly impaired in its ability to infect transformed U937 cells, a human macrophagelike cell line; i.e., the bacterial inoculum of the mutant strain that was required to initiate infection of the macrophage monolayer was ca. 80-fold greater than that of the isogenic parent strain. The mutant strain regained full infectivity on reintroduction of a cloned 24-kDa protein gene, indicating that the reduced infectivity was due specifically to the mutation in that gene. Compared with the parent strain, the mutant strain was recovered at titers that were ca. 10-fold lower shortly after infection, but it exhibited a similar intracellular growth rate over the next 40 h, indicating that the mutant was defective in its ability to initiate macrophage infection rather than in its ability to replicate intracellularly. When opsonized, the mutant strain was still significantly less infectious than the parent strain, despite equivalent macrophage association, suggesting that the mutant was not merely missing a ligand for macrophage attachment. The mutant also exhibited reduced infectivity in explanted human alveolar macrophages, demonstrating the relevance of the U937 cell model for analyzing this mutant phenotype. These results represent the first identification of a cloned L. pneumophila gene that is necessary for optimal intracellular infection; we designate this gene mip, for macrophage infectivity potentiator.

Five cases of Legionnaires' disease caused by Legionella dumoffii were identified within an 11-month period in a hospital in the Quebec City area. In four cases bacterial isolates were obtained from clinical specimens, and in one case seroconversion was demonstrated. All the patients had been admitted to hospital within 10 days before diagnosis. Two of the patients were immunosuppressed children. Only 1 of the 40 hot water samples from the hospital yielded L. dumoffii; however, 6 of 11 distilled water samples contained the bacterium. All the patients had been exposed to distilled water, four through respiratory therapy equipment and one through a room humidifier. Following the use of sterile distilled water in the apparatus, no further cases were identified. This is the first reported outbreak of Legionnaires' disease caused by L. dumoffii, and it is the first time that nosocomial legionellosis has been linked to contaminated distilled water in Canada.

Following isolation of Legionella pneumophila from a special dental station water circuit, used primarily to cool high-speed dental drills which produce fine aerosols, a case finding and environmental survey was undertaken. Widespread colonization of the dental stations was found and the results suggested that amplification of the background levels of L. pneumophila was taking place within the stations. However there was no evidence for transmission causing human infection.

In 1 year 12 of 48 patients who developed fatal pneumonia following admission with non-respiratory disorders to the Hospital Molinette, Torino, yielded Legionella pneumophila serogroup 1 from lung at autopsy. Patients were hospitalized on seven different wards for different conditions; only two of the wards had air conditioning but legionellas were not isolated from these. All patients were in poor health or immunocompromised. Some patients had inhaled humidified oxygen from piped supplies and three had undergone surgery. Legionella pneumophila serogroup 1 was detected in the water of oxygen bubble humidifiers and an underwater chest drain. The contaminated devices had been filled with tap or distilled water and the hospital water supply was found to be contaminated with L. pneumophila serogroup 1. Our findings suggest that filling bubble humidifiers or underwater chest drains with tap water is a potential hazard and should be avoided.

In addition to providing a powerful approach for identifying bacterial factors required for full infectivity and disease production, genetic analysis of Legionella pathogenesis should also lend critical insight into the biology of the macrophage and into the pathogenesis of other intracellular parasites. The interaction between L. pneumophila and the macrophage exhibits many features found in a wide variety of prokaryotic and eukaryotic intracellular human pathogens. For example, binding to complement receptors has been shown to occur for Mycobacterium tuberculosis, M. leprae, Leishmania donovani, Leishmania major and Histoplasma capsulatum. Coiling phagocytosis has been observed during entry of L. donovani. Phagosomes that contain Toxoplasma gondii or M. tuberculosis fail to fuse with lysosomes and, in the case of T. gondii, have been shown to remain close to neutral pH. Although the molecular bases for these phenomena are unknown, their functional similarities to the L. pneumophila-macrophage interaction provide optimism that generally applicable principles are involved. The genetic techniques reviewed here will provide the molecular tools with which such questions of a general biologic nature can be framed and eventually answered. Together with more traditional methods in biochemistry, microbiology and cell biology, molecular genetics offers a robust means toward identifying and understanding the bacterial factors involved in the pathogenesis of Legionnaires' disease. Molecular studies of L. pneumophila can also help address questions concerning the epidemiology, diagnosis and prevention of disease. For example, the distribution of virulence factors might help explain and predict the attack rates of different L. pneumophila strains or Legionella species. Moreover, bacterial genes/factors that are shown to be conserved in Legionella strains could be used to develop such diagnostic tools as DNA probes. Novel types of vaccines consisting of genetically constructed, avirulent L. pneumophila strains or subunit vaccines based on the molecular characterization of virulence factors might be developed and tested as protective immunogens. In this way, the capacity to analyze and to manipulate L. pneumophila genetically may facilitate the use of Legionnaires' disease as a model infection for studying protective cell-mediated immunity. Apart from its clinical significance as the etiologic agent of Legionnaires' disease, L. pneumophila may be a key to broader understandings in microbial pathogenesis and human cell biology and immunology. Although the extremely complex processes of bacterial infection and virulence are best understood when a variety of experimental approaches are employed, we believe that the evolving molecular genetic techniques reviewed here will be critical elements in many important breakthroughs in the future.

Legionella pneumophila serogroup 1 was isolated from post mortem specimens from 13 out of 58 patients with pneumonia diagnosed at autopsy. The results of a study undertaken in the hospital environment showed that the water plumbing system was colonized with L. pneumophila serogroup 1 which could also be isolated from respiratory devices filled with tap water. Control measures instituted are described.

An epidemiological link was found between contamination of a hospital water-supply by Legionella pneumophila and by Pittsburgh pneumonia agent (PPA) and subsequent cases of nosocomial legionnaires' disease and Pittsburgh pneumonia. The extent of L pneumophila isolation from the water-supply paralleled the occurrence of disease. Whenever L pneumophila was isolated from more than 30% of ten selected water sites, nosocomial legionellosis occurred. The temperature of the hot water tanks was raised to 60-77 degrees C for 72 h, and water outlets were flushed for 30 min with hot water. A decline in numbers of L pneumophila and PPA in the water-supply was followed by a fall in the incidence of legionnaires' disease and Pittsburgh pneumonia. In addition, intermittent raising of the temperature in the hot water system decreased both the number of months in which disease occurred and the proportion of nosocomial pneumonias caused by these organisms.

The etiological structure of acute pneumonia and acute respiratory diseases was studied with a view to establishing the proportion of L. pneumophila among other causative agents of such diseases. A total of 299 patients were examined over time. The etiological diagnosis based on the data of serological examination was made in 70.6% of the patients with acute pneumonia and in 65% of the patients with acute respiratory viral infections and influenza. In the etiology of pneumonia, the leading role was found to belong to influenza A (H3N2) and B viruses, as well as to adenovirus, while in the etiology of acute respiratory viral infections and influenza, to influenza B virus, adenovirus and Mycoplasma pneumoniae. The importance of L. pneumophila in the etiology of acute pneumonia and acute respiratory diseases was shown. The proportion of L. pneumophila proved to be, on the average, 9.9% in acute pneumonia and 9.8% in acute respiratory diseases. L. pneumophila occurred most frequently in mixed infections in combination with adenovirus and influenza B virus. Diseases of Legionella etiology were found to have a seasonal character, occurring mostly in winter and spring.

Six discrete protease activities were recovered from the supernatant broth of Legionella pneumophila cultures by ion-exchange chromatography. One of these demonstrated in vitro activity against collagen, casein and gelatin. When administered into the lungs of guinea-pigs this protease elicited lesions which were pathologically similar to those seen in clinical and experimentally induced Legionnaires' disease.