Pathogens

2024

Jump to: 2022, 2021, 2020, 2019

10 pages, 234 KiB

Open AccessArticle

Evaluation of Three Different Selective Media for Enumeration of Clostridium perfringens in Untreated and Treated Wastewater

by A. J. Karon, Emily S. Bailey and Mark D. Sobsey

Pathogens 2024, 13(7), 526; https://doi.org/10.3390/pathogens13070526 - 21 Jun 2024

Cited by 1 | Viewed by 962

Abstract

Current and emerging legislation in North Carolina and other regions calls for the enumeration of Clostridium perfringens as a surrogate indicator for protozoan parasites in various types of waters. Past studies that have evaluated selective media for the detection of this bacterium have [...] Read more.

Current and emerging legislation in North Carolina and other regions calls for the enumeration of Clostridium perfringens as a surrogate indicator for protozoan parasites in various types of waters. Past studies that have evaluated selective media for the detection of this bacterium have provided limited, conflicting, and inconclusive results. In this study, membrane filtration was used to enumerate C. perfringens as culturable spores or total culturable cells in 19 samples of untreated and 25 samples of partially treated wastewaters on 3 candidate media, Tryptose Sulfite Cycloserine Agar (TSC), CP ChromoSelect Agar (CPCS), and membrane Clostridium perfringens Agar (m-CP) in parallel, and the results were compared. Presumptive isolates from each agar were further subjected to phenotypic confirmation tests for acid phosphatase production and stormy fermentation to further determine the performance of each agar. The CPCS agar was determined to have the highest enumerative capacity of total C. perfringens cells when compared to both TSC agar and m-CP agar (p-value < 0.05), but there was no significant difference in its ability to detect spores when compared to TSC agar (p-value > 0.05). The overall specificity of CPCS agar as determined by agreement of results from both confirmation tests was 0.81, while the specificity of TSC agar was only 0.28. Based on its performance, ease of preparation and use and consistency of colony characteristics, CPCS agar is recommended as the preferred medium for C. perfringens enumeration in wastewater. Full article

2022

Jump to: 2024, 2021, 2020, 2019

15 pages, 1625 KiB

Open AccessArticle

The Geological Characteristics of the Vadose Zone Influence the Impact of Treated Wastewater on the Groundwater Quality (SCA.Re.S. Project 2019–2020)

by Osvalda De Giglio, Francesco Triggiano, Francesca Apollonio, Chrysovalentinos Pousis, Carla Calia, Giusy Diella, Francesco Bagordo, Sapia Murgolo, Tiziana Grassi, Cristina De Ceglie, Silvia Brigida, Giuseppina La Rosa, Pamela Mancini, Giusy Bonanno Ferraro, Antonella De Donno, Giuseppe Mascolo, Maria Clementina Caputo and Maria Teresa Montagna

Pathogens 2022, 11(6), 677; https://doi.org/10.3390/pathogens11060677 - 11 Jun 2022

Cited by 3 | Viewed by 2192

Abstract

This study evaluated whether some chemical and microbial contaminants in treated sewage effluents from two wastewater treatment plants (WWTP) reached the groundwater when they drained through a fractured karst vadose zone (WWTP-K) and a porous vadose zone (WWTP-P). Forty-five samples of sewage water [...] Read more.

This study evaluated whether some chemical and microbial contaminants in treated sewage effluents from two wastewater treatment plants (WWTP) reached the groundwater when they drained through a fractured karst vadose zone (WWTP-K) and a porous vadose zone (WWTP-P). Forty-five samples of sewage water (SW), treated water (TW), and monitoring well (MW), collected from WWTP-P (24) and WWTP-K (21), were analyzed for a range of microbiological and chemical properties. The E. coli and Salmonella counts were below the limits outlined in the Legislative Decree 152/06 in effluents from both types of WWTP. Enteric viruses were found in 37.5% and 12.5% of the SW and TW from WWTP-P, respectively. The percentages of Pepper mild mottle virus isolated were higher in TW (62.5% in WWTP-P, 85.7% in WWTP-K) than in SW and MW. The residual concentrations of contaminants of emerging concern (CEC) of each drug category were higher in the MW downstream of WWTP-K than of WWTP-P. Our results showed that the porous vadose zone was more effective at reducing the contaminant loads than the fractured karst one, especially the CEC, in the effluent. The legislation should include other parameters to minimize the risks from treated effluent that is discharged to soil. Full article

► Show Figures

Figure 1

13 pages, 846 KiB

Open AccessArticle

Irrigation of Olives with Reclaimed Wastewaters and Deficit Strategies Affect Pathogenic Bacteria Contamination of Water and Soil

by Gaetano Alessandro Vivaldi, Salvatore Camposeo, Gabriele Caponio, Giuseppe Lopriore, Francesco Discipio, Francesca Apollonio, Francesco Triggiano, Osvalda De Giglio and Maria Teresa Montagna

Pathogens 2022, 11(5), 488; https://doi.org/10.3390/pathogens11050488 - 20 Apr 2022

Cited by 3 | Viewed by 2242

Abstract

This study aimed to evaluate pathogenic bacterial contamination of the water-soil-plant system in potted olive trees irrigated with reclaimed wastewater. Desalinated water (DW) obtained by treating municipal wastewater (SW) and reclaimed water (RW) obtained by mixing SW with the brine (BR) produced by [...] Read more.

This study aimed to evaluate pathogenic bacterial contamination of the water-soil-plant system in potted olive trees irrigated with reclaimed wastewater. Desalinated water (DW) obtained by treating municipal wastewater (SW) and reclaimed water (RW) obtained by mixing SW with the brine (BR) produced by DESERT technology (tertiary treatment by ultrafiltration, active carbon and reverse osmosis) were used. Two different irrigation regimes were compared: full irrigation (FI) and regulated deficit irrigation (RDI). During two irrigation seasons the concentrations of Escherichia coli, enterococci, spores of sulfite-reducing Clostridia (SRC) and Salmonella spp. were monitored in water, soil and fruit samples. Microbial concentrations in DW were always below the threshold for reuse in agriculture, while RW showed the highest level of contamination for all observed parameters. RDI management appeared to increase the soil content of SRC spores with respect to FI. Sporadically low SRC spore contamination was recorded in some fruits only in 2018, regardless of the irrigation source, probably because of accidental contamination during sampling or post-harvest handling. This study encourages the creation of a better regulatory framework reference, with specific guidelines for the use of RW as part of integrated environmental systems for the management of sustainable agriculture. Full article

► Show Figures

Figure 1

8 pages, 227 KiB

Open AccessArticle

Desiccation-Tolerance of Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium chimaera, Mycobacterium abscessus and Mycobacterium chelonae

by Joseph O. Falkinham III and Myra D. Williams

Pathogens 2022, 11(4), 463; https://doi.org/10.3390/pathogens11040463 - 13 Apr 2022

Cited by 3 | Viewed by 1844

Abstract

Desiccation-tolerance of cells of four strains of Mycobacterium chimaera and individual strains of Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, and Mycobacterium chelonae were measured by two methods. The survival of water-acclimated cells both in filter paper and on the surface [...] Read more.

Desiccation-tolerance of cells of four strains of Mycobacterium chimaera and individual strains of Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, and Mycobacterium chelonae were measured by two methods. The survival of water-acclimated cells both in filter paper and on the surface of stainless-steel coupons were measured. In filter paper at 40% relative humidity at 25 °C, survival of patient isolates of M. avium and M. chimaera cells was 28% and 34% after 21 days of incubation, whereas it was 100% for the Sorin 3T isolate of M. chimaera. On stainless-steel biofilms after 42 days of incubation at 40% relative humidity at 25 °C, survival of water-acclimated cells of M. intracellulare was above 100%, while M. chelonae cells did not survive beyond 21 days, and survival of water-acclimated cells of M. avium and M. abscessus was 18% and 14%, respectively. On stainless-steel coupons, survival of patient and Sorin 3T isolates of M. chimaera was quite similar, specifically between 14% and 28% survival, after 42 days of incubation at 40% relative humidity at 25 °C. The experiments would support the hypothesis that some nontuberculous mycobacterial species are relatively desiccation-tolerant, whereas others are not. Further, long-term survival of the two M. chimaera strains is consistent with the presence of that species in Sorin 3T heater-coolers shipped throughout the world. Full article

8 pages, 1870 KiB

Open AccessArticle

Low Efficacy of Periodical Thermal Shock for Long-Term Control of Legionella spp. in Hot Water System of Hotels

by Jhon J. Molina, Magdalena Bennassar, Edwin Palacio and Sebastian Crespi

Pathogens 2022, 11(2), 152; https://doi.org/10.3390/pathogens11020152 - 25 Jan 2022

Cited by 7 | Viewed by 3229

Abstract

Different guidelines and regulations for the prevention of legionellosis in public facilities include the recommendation of a periodical thermal shock in the hot water system. The purpose of this study was to assess the efficacy of periodical thermal shocks along a 1-year period [...] Read more.

Different guidelines and regulations for the prevention of legionellosis in public facilities include the recommendation of a periodical thermal shock in the hot water system. The purpose of this study was to assess the efficacy of periodical thermal shocks along a 1-year period on the presence of Legionella spp. in the domestic hot water system of hotels. The Legionella testing results from the period January–December 2019 coming from a group of 77 hotel facilities in the Balearic Islands (Spain) conducting periodical thermal shocks were analyzed. A second group of 44 hotels operating without periodical thermal shocks was used for a comparative analysis. In the facilities where the periodical thermal shock was performed, 16.0% of the results (429 hot water samples collected) were positive for Legionella spp., compared to 21.1% (298 samples), where periodical thermal shock was not performed. Overall, in the thermal shock group, 32.5% of the sites presented at least 1 positive sample along the period of study versus 45.5% in the control group. None of these differences was statistically significant (p-value > 0.05). These findings suggest that the efficacy of regular thermal shock for long-term control of Legionella spp. in domestic hot water systems of hotels is low. Full article

► Show Figures

Figure 1

2021

Jump to: 2024, 2022, 2020, 2019

15 pages, 3834 KiB

Open AccessArticle

Occurrence of Legionella spp. in Man-Made Water Sources: Isolates Distribution and Phylogenetic Characterization in the Emilia-Romagna Region

by Marta Mazzotta, Silvano Salaris, Maria Rosaria Pascale, Luna Girolamini and Sandra Cristino

Pathogens 2021, 10(5), 552; https://doi.org/10.3390/pathogens10050552 - 3 May 2021

Cited by 16 | Viewed by 2664

Abstract

Legionella species distribution in the Emilia-Romagna region, involving hospital (H) and community (C) environments, was conducted. Legionella culture, agglutination test, and mip-gene sequencing were applied on 240 isolates. The analysis showed a higher prevalence of non-Legionellapneumophila (n-Lp) species [...] Read more.

Legionella species distribution in the Emilia-Romagna region, involving hospital (H) and community (C) environments, was conducted. Legionella culture, agglutination test, and mip-gene sequencing were applied on 240 isolates. The analysis showed a higher prevalence of non-Legionellapneumophila (n-Lp) species (84.1%) compared with L. pneumophila (Lp) (15.9%), with a higher frequency of n-Lp with respect to Lp species in both environments (77.6% and 96.4%, in H and C, respectively). The Shannon index showed a significant difference in Legionella distribution (p = 0.00017), with a significant abundance of Lp in the H compared with C environment (p = 0.00028). The continuous disinfection treatment in H could contribute to adaptive survival of the Lp species. Phylogenetic analysis revealed a conservative clade distribution between H and C: L. feeleii clade with three subclades in C and the Lp clade with five subclades in H and two in C, respectively. Our findings suggest the importance of Legionella surveillance both in H and C, with a focus on n-Lp species less connected to human disease. The Legionella prevalence and diversity found here indicate that geographical and temporal isolate evolution should be considered during surveillance, particularly in the light of global warming and changes in population risk factors. Full article

► Show Figures

Figure 1

2020

Jump to: 2024, 2022, 2021, 2019

6 pages, 224 KiB

Open AccessEditorial

Editorial Comments to the Special Issue: “Legionella Contamination in Water Environment”

by Silvia Bonetta and Sara Bonetta

Pathogens 2020, 9(12), 1017; https://doi.org/10.3390/pathogens9121017 - 2 Dec 2020

Cited by 2 | Viewed by 1885

Abstract

Legionella spp [...] Full article

23 pages, 3778 KiB

Open AccessArticle

Biogeography and Environmental Drivers of Legionella pneumophila Abundance and Genotype Composition across the West Bank: Relevance of a Genotype-Based Ecology for Understanding Legionella Occurrence

by Ashraf R. Zayed, Suha Butmeh, Marina Pecellin, Alaa Salah, Hanna Alalam, Michael Steinert, Manfred G. Höfle, Dina M. Bitar and Ingrid Brettar

Pathogens 2020, 9(12), 1012; https://doi.org/10.3390/pathogens9121012 - 1 Dec 2020

Cited by 11 | Viewed by 2460

Abstract

The West Bank can be considered as a high-risk area for Legionella prevalence in drinking water due to high ambient temperature, intermittent water supply, frequent pressure loss, and storage of drinking water in roof containers. To assess occurrence of Legionella species, especially L. [...] Read more.

The West Bank can be considered as a high-risk area for Legionella prevalence in drinking water due to high ambient temperature, intermittent water supply, frequent pressure loss, and storage of drinking water in roof containers. To assess occurrence of Legionella species, especially L. pneumophila, in the drinking water of the West Bank, the drinking water distribution systems of eight hospitals were sampled over a period of 2.3 years covering the seasonal cycle and the major geographic regions. To gain insight into potential environmental drivers, a set of physico-chemical and microbiological parameters was recorded. Sampling included drinking water and biofilm analyzed by culture and PCR-based methods. Cultivation led to the isolation of 180 strains of L. pneumophila that were genotyped by Multi-Locus Variable Number of Tandem Repeat Analysis (MLVA). Surprisingly, the abundance of culturable L. pneumophila was low in drinking water of the sampling sites, with only three out of eight sites where Legionella was observed at all (range: 30–500 CFU/Liter). By contrast, biofilm and PCR-based analyses showed a higher prevalence. Statistical analyses with physico-chemical parameters revealed a decrease of L. pneumophila abundance for water and biofilm with increasing magnesium concentrations (>30 mg/L). MLVA-genotype analysis of the L. pneumophila isolates and their spatial distribution indicated three niches characterized by distinct physico-chemical parameters and inhabited by specific consortia of genotypes. This study provides novel insights into mechanisms shaping L. pneumophila populations and triggering their abundance leading to an understanding of their genotype-specific niches and ecology in support of improved prevention measures. Full article

► Show Figures

Figure 1

34 pages, 2696 KiB

Open AccessEditor’s ChoiceReview

Critical Review: Propensity of Premise Plumbing Pipe Materials to Enhance or Diminish Growth of Legionella and Other Opportunistic Pathogens

by Abraham C. Cullom, Rebekah L. Martin, Yang Song, Krista Williams, Amanda Williams, Amy Pruden and Marc A. Edwards

Pathogens 2020, 9(11), 957; https://doi.org/10.3390/pathogens9110957 - 17 Nov 2020

Cited by 47 | Viewed by 8318

Abstract

Growth of Legionella pneumophila and other opportunistic pathogens (OPs) in drinking water premise plumbing poses an increasing public health concern. Premise plumbing is constructed of a variety of materials, creating complex environments that vary chemically, microbiologically, spatially, and temporally in a manner likely [...] Read more.

Growth of Legionella pneumophila and other opportunistic pathogens (OPs) in drinking water premise plumbing poses an increasing public health concern. Premise plumbing is constructed of a variety of materials, creating complex environments that vary chemically, microbiologically, spatially, and temporally in a manner likely to influence survival and growth of OPs. Here we systematically review the literature to critically examine the varied effects of common metallic (copper, iron) and plastic (PVC, cross-linked polyethylene (PEX)) pipe materials on factors influencing OP growth in drinking water, including nutrient availability, disinfectant levels, and the composition of the broader microbiome. Plastic pipes can leach organic carbon, but demonstrate a lower disinfectant demand and fewer water chemistry interactions. Iron pipes may provide OPs with nutrients directly or indirectly, exhibiting a high disinfectant demand and potential to form scales with high surface areas suitable for biofilm colonization. While copper pipes are known for their antimicrobial properties, evidence of their efficacy for OP control is inconsistent. Under some circumstances, copper’s interactions with premise plumbing water chemistry and resident microbes can encourage growth of OPs. Plumbing design, configuration, and operation can be manipulated to control such interactions and health outcomes. Influences of pipe materials on OP physiology should also be considered, including the possibility of influencing virulence and antibiotic resistance. In conclusion, all known pipe materials have a potential to either stimulate or inhibit OP growth, depending on the circumstances. This review delineates some of these circumstances and informs future research and guidance towards effective deployment of pipe materials for control of OPs. Full article

► Show Figures

Figure 1

24 pages, 1648 KiB

Open AccessEditor’s ChoiceArticle

Characterization of Legionella pneumophila Populations by Multilocus Variable Number of Tandem Repeats (MLVA) Genotyping from Drinking Water and Biofilm in Hospitals from Different Regions of the West Bank

by Ashraf R. Zayed, Marina Pecellin, Alaa Salah, Hanna Alalam, Suha Butmeh, Michael Steinert, Rene Lesnik, Ingrid Brettar, Manfred G. Höfle and Dina M. Bitar

Pathogens 2020, 9(11), 862; https://doi.org/10.3390/pathogens9110862 - 22 Oct 2020

Cited by 10 | Viewed by 3919

Abstract

The West Bank can be considered a high-risk area for Legionnaires’ disease (LD) due to its hot climate, intermittent water supply and roof storage of drinking water. Legionella, mostly L. pneumophila, are responsible for LD, a severe, community-acquired and nosocomial pneumonia. [...] Read more.

The West Bank can be considered a high-risk area for Legionnaires’ disease (LD) due to its hot climate, intermittent water supply and roof storage of drinking water. Legionella, mostly L. pneumophila, are responsible for LD, a severe, community-acquired and nosocomial pneumonia. To date, no extensive assessment of Legionella spp and L. pneumophila using cultivation in combination with molecular approaches in the West Bank has been published. Two years of environmental surveillance of Legionella in water and biofilms in the drinking water distribution systems (DWDS) of eight hospitals was carried out; 180 L. pneumophila strains were isolated, mostly from biofilms in DWDS. Most of the isolates were identified as serogroup (Sg) 1 (60%) and 6 (30%), while a minor fraction comprised Sg 8 and 10. Multilocus Variable number of tandem repeats Analysis using 13 loci (MLVA-8(12)) was applied as a high-resolution genotyping method and compared to the standard Sequence Based Typing (SBT). The isolates were genotyped in 27 MLVA-8(12) genotypes (Gt), comprising four MLVA clonal complexes (VACC 1; 2; 5; 11). The major fraction of isolates constituted Sequence Type (ST)1 and ST461. Most of the MLVA-genotypes were highly diverse and often unique. The MLVA-genotype composition showed substantial regional variability. In general, the applied MLVA-method made it possible to reproducibly genotype the isolates, and was consistent with SBT but showed a higher resolution. The advantage of the higher resolution was most evident for the subdivision of the large strain sets of ST1 and ST461; these STs were shown to be highly pneumonia-relevant in a former study. This shows that the resolution by MLVA is advantageous for back-tracking risk sites and for the avoidance of outbreaks of L. pneumophila. Overall, our results provide important insights into the detailed population structure of L. pneumophila, allowing for better risk assessment for DWDS. Full article

► Show Figures

Graphical abstract

6 pages, 439 KiB

Open AccessArticle

Evaluation of GVPC and BCYE Media for Legionella Detection and Enumeration in Water Samples by ISO 11731: Does Plating on BCYE Medium Really Improve Yield?

by Maria Scaturro, Elisa Poznanski, Mariarosaria Mupo, Paola Blasior, Margit Seeber, Anna-Maria Prast, Elisa Romanin, Antonietta Girolamo, Maria Cristina Rota, Antonino Bella, Maria Luisa Ricci and Alberta Stenico

Pathogens 2020, 9(9), 757; https://doi.org/10.3390/pathogens9090757 - 16 Sep 2020

Cited by 10 | Viewed by 4812

Abstract

Legionella spp are the causative agents of Legionnaires’ diseases, which is a pneumonia of important public health concern. Ubiquitous freshwater and soil inhabitants can reach man-made water systems and cause illness. Legionella enumeration and quantification in water systems is crucial for risk assessment [...] Read more.

Legionella spp are the causative agents of Legionnaires’ diseases, which is a pneumonia of important public health concern. Ubiquitous freshwater and soil inhabitants can reach man-made water systems and cause illness. Legionella enumeration and quantification in water systems is crucial for risk assessment and culture examination is the gold standard method. In this study, Legionella recovery from potable water samples, at presumably a low concentration of interfering microorganisms, was compared by plating on buffered charcoal yeast extract (BCYE) and glycine, vancomycin, polymyxin B, cycloheximide (GVPC) Legionella agar media, according to the International Standard Organization (ISO) 11731: 2017. Overall, 556 potable water samples were analyzed and 151 (27.1%) were positive for Legionella. Legionella grew on both BCYE and GVPC agar plates in 85/151 (56.3%) water samples, in 65/151 (43%) on only GVPC agar plates, and in 1/151 (0.7%) on only BCYE agar plates. In addition, GVPC medium identified Legionella species other than pneumophila in six more samples as compared with the culture on BCYE. Although the medians of colony forming units per liter (CFU/L) detected on the BCYE and GVPC agar plates were 2500 and 1350, respectively (p-value < 0.0001), the difference did not exceed one logarithm, and therefore is not relevant for Legionella risk assessment. These results make questionable the need to utilize BCYE agar plates to analyze potable water samples. Full article

► Show Figures

Figure 1

17 pages, 3472 KiB

Open AccessArticle

Interactive Effects of Copper Pipe, Stagnation, Corrosion Control, and Disinfectant Residual Influenced Reduction of Legionella pneumophila during Simulations of the Flint Water Crisis

by Rebekah L. Martin, Owen R. Strom, Amy Pruden and Marc A. Edwards

Pathogens 2020, 9(9), 730; https://doi.org/10.3390/pathogens9090730 - 4 Sep 2020

Cited by 23 | Viewed by 4894

Abstract

Flint, MI experienced two outbreaks of Legionnaires’ Disease (LD) during the summers of 2014 and 2015, coinciding with use of Flint River as a drinking water source without corrosion control. Using simulated distribution systems (SDSs) followed by stagnant simulated premise (i.e., building) plumbing [...] Read more.

Flint, MI experienced two outbreaks of Legionnaires’ Disease (LD) during the summers of 2014 and 2015, coinciding with use of Flint River as a drinking water source without corrosion control. Using simulated distribution systems (SDSs) followed by stagnant simulated premise (i.e., building) plumbing reactors (SPPRs) containing cross-linked polyethylene (PEX) or copper pipe, we reproduced trends in water chemistry and Legionella proliferation observed in the field when Flint River versus Detroit water were used before, during, and after the outbreak. Specifically, due to high chlorine demand in the SDSs, SPPRs with treated Flint River water were chlorine deficient and had elevated L. pneumophila numbers in the PEX condition. SPPRs with Detroit water, which had lower chlorine demand and higher residual chlorine, lost all culturable L. pneumophila within two months. L. pneumophila also diminished more rapidly with time in Flint River SPPRs with copper pipe, presumably due to the bacteriostatic properties of elevated copper concentrations caused by lack of corrosion control and stagnation. This study confirms hypothesized mechanisms by which the switch in water chemistry, pipe materials, and different flow patterns in Flint premise plumbing may have contributed to observed LD outbreak patterns. Full article

► Show Figures

Figure 1

8 pages, 241 KiB

Open AccessArticle

Performance of Legiolert Test vs. ISO 11731 to Confirm Legionella pneumophila Contamination in Potable Water Samples

by Maria Scaturro, Matteo Buffoni, Antonietta Girolamo, Sandra Cristino, Luna Girolamini, Marta Mazzotta, Maria Antonietta Bucci Sabattini, Cristina Maria Zaccaro, Leonarda Chetti, Microbiology Arpa Novara Laboratory, Antonino Bella, Maria Cristina Rota and Maria Luisa Ricci

Pathogens 2020, 9(9), 690; https://doi.org/10.3390/pathogens9090690 - 23 Aug 2020

Cited by 23 | Viewed by 3928

Abstract

Detection and enumeration of Legionella in water samples is of great importance for risk assessment analysis. The plate culture method is the gold standard, but has received several well-known criticisms, which have induced researchers to develop alternative methods. The purpose of this study [...] Read more.

Detection and enumeration of Legionella in water samples is of great importance for risk assessment analysis. The plate culture method is the gold standard, but has received several well-known criticisms, which have induced researchers to develop alternative methods. The purpose of this study was to compare Legionella counts obtained by the analysis of potable water samples through the plate culture method and through the IDEXX liquid culture Legiolert method. Legionella plate culture, according to ISO 11731:1998, was performed using 1 L of water. Legiolert was performed using both the 10 mL and 100 mL Legiolert protocols. Overall, 123 potable water samples were analyzed. Thirty-seven (30%) of them, positive for L. pneumophila, serogroups 1 or 2–14 by plate culture, were used for comparison with the Legiolert results. The Legiolert 10 mL test detected 34 positive samples (27.6%) and the Legiolert 100 mL test detected 37 positive samples, 27.6% and 30% respectively, out of the total samples analyzed. No significant difference was found between either the Legiolert 10 mL and Legiolert 100 mL vs. the plate culture (p = 0.9 and p = 0.3, respectively) or between the Legiolert 10 mL and Legiolert 100 mL tests (p = 0.83). This study confirms the reliability of the IDEXX Legiolert test for Legionella pneumophila detection and enumeration, as already shown in similar studies. Like the plate culture method, the Legiolert assay is also suitable for obtaining isolates for typing purposes, relevant for epidemiological investigations. Full article

29 pages, 2943 KiB

Open AccessEditor’s ChoiceArticle

Legionella Diversity and Spatiotemporal Variation in the Occurrence of Opportunistic Pathogens within a Large Building Water System

by Helen Y. Buse, Brian J. Morris, Vicente Gomez-Alvarez, Jeffrey G. Szabo and John S. Hall

Pathogens 2020, 9(7), 567; https://doi.org/10.3390/pathogens9070567 - 13 Jul 2020

Cited by 21 | Viewed by 4082

Abstract

Understanding Legionella survival mechanisms within building water systems (BWSs) is challenging due to varying engineering, operational, and water quality characteristics unique to each system. This study aimed to evaluate Legionella, mycobacteria, and free-living amoebae occurrence within a BWS over 18–28 months at [...] Read more.

Understanding Legionella survival mechanisms within building water systems (BWSs) is challenging due to varying engineering, operational, and water quality characteristics unique to each system. This study aimed to evaluate Legionella, mycobacteria, and free-living amoebae occurrence within a BWS over 18–28 months at six locations differing in plumbing material and potable water age, quality, and usage. A total of 114 bulk water and 57 biofilm samples were analyzed. Legionella culturability fluctuated seasonally with most culture-positive samples being collected during the winter compared to the spring, summer, and fall months. Positive and negative correlations between Legionella and L. pneumophila occurrence and other physiochemical and microbial water quality parameters varied between location and sample types. Whole genome sequencing of 19 presumptive Legionella isolates, from four locations across three time points, identified nine isolates as L. pneumophila serogroup (sg) 1 sequence-type (ST) 1; three as L. pneumophila sg5 ST1950 and ST2037; six as L. feeleii; and one as Ochrobactrum. Results showed the presence of a diverse Legionella population with consistent and sporadic occurrence at four and two locations, respectively. Viewed collectively with similar studies, this information will enable a better understanding of the engineering, operational, and water quality parameters supporting Legionella growth within BWSs. Full article

► Show Figures

Figure 1

9 pages, 236 KiB

Open AccessArticle

Sensitivity and Selectivity of Two Commercially Available Media for Legionella spp. Recovery from Environmental Water Samples

by Savina Ditommaso, Monica Giacomuzzi, Gabriele Memoli, Jacopo Garlasco and Carla M. Zotti

Pathogens 2020, 9(7), 523; https://doi.org/10.3390/pathogens9070523 - 29 Jun 2020

Cited by 3 | Viewed by 3858

Abstract

The quality control of culture media used for Legionella spp. isolation and enumeration is paramount to achieve a satisfactory degree of comparability among water testing results from different laboratories. Here, we report on a comparative assessment of the sensitivity and selectivity of MWY [...] Read more.

The quality control of culture media used for Legionella spp. isolation and enumeration is paramount to achieve a satisfactory degree of comparability among water testing results from different laboratories. Here, we report on a comparative assessment of the sensitivity and selectivity of MWY and BCYEα media supplied by two different manufacturers (i.e., Xebios Diagnostics GmbH and Oxoid Ltd) for the detection of Legionella spp. from environmental water samples. Even though our analysis showed an excellent agreement between the recovery rates of the four media tested (90.5%), the quantitative recovery of Legionella spp. colonies using Xebios media was significantly greater than that achieved by Oxoid media (P = 0.0054). Furthermore, the sensitivity of detection was significantly higher when samples were plated on MWY ^Xebios agar (P = 0.0442), while the selectivity of MWY appeared to be the same regardless of the manufacturer. Furthermore, MWY^Xebios agar favored the growth of much larger colonies compared to those observed on MWY^Oxoid agar. Finally, MWY^Xebios medium enhanced the recovery of non-pneumophila Legionella species. Collectively, our findings demonstrate that quality control is crucial to ensure high selectivity and sensitivity of the culture media used for the detection and enumeration of Legionella spp. from environmental water resources. As water remediation measures strictly depend on Legionella spp. recovery, culture protocol standardization, as well as quality control of the culture media, is essential to achieve intra- and interlaboratory reproducibility and accuracy. Full article

12 pages, 1085 KiB

Open AccessArticle

Mitigation of Expression of Virulence Genes in Legionella pneumophila Internalized in the Free-Living Amoeba Willaertia magna C2c Maky

by Rayane Mouh Mameri, Jacques Bodennec, Laurent Bezin and Sandrine Demanèche

Pathogens 2020, 9(6), 447; https://doi.org/10.3390/pathogens9060447 - 5 Jun 2020

Cited by 4 | Viewed by 2700

Abstract

Legionella pneumophila is a human pathogen responsible for a severe form of pneumonia named Legionnaire disease. Its natural habitat is aquatic environments, being in a free state or intracellular parasites of free-living amoebae, such as Acanthamoeba castellanii. This pathogen is able to [...] Read more.

Legionella pneumophila is a human pathogen responsible for a severe form of pneumonia named Legionnaire disease. Its natural habitat is aquatic environments, being in a free state or intracellular parasites of free-living amoebae, such as Acanthamoeba castellanii. This pathogen is able to replicate within some amoebae. Willaertia magna C2c Maky, a non-pathogenic amoeba, was previously demonstrated to resist to L. pneumophila and even to be able to eliminate the L. pneumophila strains Philadelphia, Lens, and Paris. Here, we studied the induction of seven virulence genes of three L. pneumophila strains (Paris, Philadelphia, and Lens) within W. magna C2c Maky in comparison within A. castellanii and with the gene expression level of L. pneumophila strains alone used as controls. We defined a gene expression-based virulence index to compare easily and without bias the transcript levels in different conditions and demonstrated that W. magna C2c Maky did not increase the virulence of L. pneumophila strains in contrast to A. castellanii. These results confirmed the non-permissiveness of W. magna C2c Maky toward L. pneumophila strains. Full article

► Show Figures

Figure 1

17 pages, 1838 KiB

Open AccessArticle

The Role of Sensor-Activated Faucets in Surgical Handwashing Environment as a Reservoir of Legionella

by Marta Mazzotta, Luna Girolamini, Maria Rosaria Pascale, Jessica Lizzadro, Silvano Salaris, Ada Dormi and Sandra Cristino

Pathogens 2020, 9(6), 446; https://doi.org/10.3390/pathogens9060446 - 5 Jun 2020

Cited by 6 | Viewed by 3223

Abstract

Surgical handwashing is a mandatory practice to protect both surgeons and patients in order to control Healthcare-Associated Infections (HAIs). The study is focused on Legionella and Pseudomonas aeruginosa contamination in Surgical Handwashing Outlets (SHWOs) provided by sensor-activated faucets with Thermostatic Mixer Valves (TMVs), [...] Read more.

Surgical handwashing is a mandatory practice to protect both surgeons and patients in order to control Healthcare-Associated Infections (HAIs). The study is focused on Legionella and Pseudomonas aeruginosa contamination in Surgical Handwashing Outlets (SHWOs) provided by sensor-activated faucets with Thermostatic Mixer Valves (TMVs), as correlated to temperature, technologies, and disinfection used. Samples were analyzed by standard culture techniques, comparing hot- and cold-water samples. Legionella isolates were typed by an agglutination test and by mip sequencing. Legionella contamination showed the same distribution between hot and cold samples concerning positive samples and mean concentration: 44.5% and 1.94 Log₁₀ cfu/L vs. 42.6% and 1.81 Log₁₀ cfu/L, respectively. Regarding the distribution of isolates (Legionella pneumophila vs. Legionella non-pneumophila species), significant differences were found between hot- and cold-positive samples. The contamination found in relation to ranges of temperature showed the main positive samples (47.1%) between 45.1–49.6 °C, corresponding to high Legionella concentrations (2.17 Log₁₀ cfu/L). In contrast, an increase of temperature (>49.6 °C) led to a decrease in positive samples (23.2%) and mean concentration (1.64 Log₁₀ cfu/L). A low level of Pseudomonas aeruginosa was found. For SHWOs located in critical areas, lack of consideration of technologies used and uncorrected disinfection protocols may lead to the development of a high-risk environment for both patients and surgeons. Full article

► Show Figures

Graphical abstract

14 pages, 879 KiB

Open AccessArticle

A Snapshot of the Prevalence and Molecular Diversity of Legionella pneumophila in the Water Systems of Israeli Hotels

by Eugenia Yakunin, Eszter Kostyal, Vered Agmon, Itamar Grotto, Lea Valinsky and Jacob Moran-Gilad

Pathogens 2020, 9(6), 414; https://doi.org/10.3390/pathogens9060414 - 27 May 2020

Cited by 14 | Viewed by 2690

Abstract

Exposure to Legionella spp. contaminated aerosols in hotel settings confers risk for travel-associated Legionnaire’s disease (TALD). In this study, we investigated the prevalence of Legionella contamination and its molecular diversity in hotels and resorts across Israel. The study was comprised of a convenience [...] Read more.

Exposure to Legionella spp. contaminated aerosols in hotel settings confers risk for travel-associated Legionnaire’s disease (TALD). In this study, we investigated the prevalence of Legionella contamination and its molecular diversity in hotels and resorts across Israel. The study was comprised of a convenience sample of water systems from 168 hotels and resorts countrywide, routinely inspected between March 2015 and February 2017. Isolation and quantitation of Legionella were performed in a water laboratory using the ISO 11731 method. The distribution of Legionella isolates was analyzed according to geography and source. The genetic diversity of a subset of isolates was analyzed by sequence-based typing (SBT) at the National Reference Laboratory for Legionella and compared to the national database. Out of 2830 samples tested, 470 (17%) obtained from 102 different premises (60% of hotels) were positive for Legionella spp. In 230 samples (49% of all positive, 8% of total samples), accounting for 37% of hotels, Legionella spp. counts exceeded the regulatory threshold of 1000 CFU/L. The most frequently contaminated water sources were cooling towers (38%), followed by faucets, hot tubs, water lines, and storage tanks (14–17% each). Furthermore, 32% and 17% of samples obtained from cooling towers and hot tubs, respectively, exceeded the regulatory thresholds. SBT was performed on 78 strains and revealed 27 different sequence types (STs), including two novel STs. The most prevalent STs found were ST1 (26%), ST87 (10%), ST93 (6%), and ST461 and ST1516 (5% each). Several L. pneumophila STs were found to be limited to certain geographical regions. This is the first study to investigate the prevalence and diversity of Legionella in hotels and resorts in Israel during non-outbreak environmental inspections. These findings will inform risk assessment, surveillance, and control measures of TALD. Full article

► Show Figures

Figure 1

12 pages, 409 KiB

Open AccessEditor’s ChoiceArticle

Prevention and Control of Legionella and Pseudomonas spp. Colonization in Dental Units

by Benedetta Tuvo, Michele Totaro, Maria Luisa Cristina, Anna Maria Spagnolo, David Di Cave, Sara Profeti, Angelo Baggiani, Gaetano Privitera and Beatrice Casini

Pathogens 2020, 9(4), 305; https://doi.org/10.3390/pathogens9040305 - 21 Apr 2020

Cited by 29 | Viewed by 4009

Abstract

Introduction: Dental Unit Waterlines (DUWLs) have shown to be a source of Legionella infection. We report the experience of different dental healthcare settings where a risk management plan was implemented. Materials and methods: In a Hospital Odontostomatology Clinic (HOC) and three Private Dental [...] Read more.

Introduction: Dental Unit Waterlines (DUWLs) have shown to be a source of Legionella infection. We report the experience of different dental healthcare settings where a risk management plan was implemented. Materials and methods: In a Hospital Odontostomatology Clinic (HOC) and three Private Dental Clinics (PDCs) housing 13 and six dental units (DUs), respectively, an assessment checklist was applied to evaluate staff compliance with guideline recommendations. DUWLs microbial parameters were investigated before and after the application of corrective actions. Results: In the HOC a poor adherence to good practices was demonstrated, whereas protocols were carefully applied in PDCs. L. pneumophila sg 2–15 was isolated in 31% (4/13) and 33% (2/6) of DUs in HOC and PDCs, respectively, mainly from handpieces (32%, 6/19) with counts >10² colony-forming units per milliliter (CFU/L), often associated with P. aeruginosa (68%, 13/19). The shock disinfection with 3% v/v hydrogen peroxide (HP) showed a limited effect, with a recolonization period of about 4 weeks. Legionella was eradicated only after 6% v/v HP shock disinfection and filters-installation, whilst P. aeruginosa after the third shock disinfection with a solution of 4% v/v HP and biodegradable surfactants. Conclusions: Our data demonstrate the presence and persistence of microbial contamination within the DUWLs, which required strict adherence to control measures and the choice of effective disinfectants. Full article

► Show Figures

Figure 1

23 pages, 487 KiB

Open AccessEditor’s ChoiceReview

Legionella pneumophila and Protozoan Hosts: Implications for the Control of Hospital and Potable Water Systems

by Muhammad Atif Nisar, Kirstin E. Ross, Melissa H. Brown, Richard Bentham and Harriet Whiley

Pathogens 2020, 9(4), 286; https://doi.org/10.3390/pathogens9040286 - 15 Apr 2020

Cited by 33 | Viewed by 4992

Abstract

Legionella pneumophila is an opportunistic waterborne pathogen of public health concern. It is the causative agent of Legionnaires’ disease (LD) and Pontiac fever and is ubiquitous in manufactured water systems, where protozoan hosts and complex microbial communities provide protection from disinfection procedures. This [...] Read more.

Legionella pneumophila is an opportunistic waterborne pathogen of public health concern. It is the causative agent of Legionnaires’ disease (LD) and Pontiac fever and is ubiquitous in manufactured water systems, where protozoan hosts and complex microbial communities provide protection from disinfection procedures. This review collates the literature describing interactions between L. pneumophila and protozoan hosts in hospital and municipal potable water distribution systems. The effectiveness of currently available water disinfection protocols to control L. pneumophila and its protozoan hosts is explored. The studies identified in this systematic literature review demonstrated the failure of common disinfection procedures to achieve long term elimination of L. pneumophila and protozoan hosts from potable water. It has been demonstrated that protozoan hosts facilitate the intracellular replication and packaging of viable L. pneumophila in infectious vesicles; whereas, cyst-forming protozoans provide protection from prolonged environmental stress. Disinfection procedures and protozoan hosts also facilitate biogenesis of viable but non-culturable (VBNC) L. pneumophila which have been shown to be highly resistant to many water disinfection protocols. In conclusion, a better understanding of L. pneumophila-protozoan interactions and the structure of complex microbial biofilms is required for the improved management of L. pneumophila and the prevention of LD. Full article

► Show Figures

Figure 1

15 pages, 3498 KiB

Open AccessArticle

Intracellular Behaviour of Three Legionella pneumophila Strains within Three Amoeba Strains, Including Willaertia magna C2c Maky

by Issam Hasni, Antoine Jarry, Benjamin Quelard, Antoine Carlino, Jean-Baptiste Eberst, Olivier Abbe and Sandrine Demanèche

Pathogens 2020, 9(2), 105; https://doi.org/10.3390/pathogens9020105 - 6 Feb 2020

Cited by 13 | Viewed by 4432

Abstract

Legionella pneumophila is a facultative intracellular pathogen found in aquatic environments as planktonic cells within biofilms and as intracellular parasites of free-living amoebae such as Acanthamoeba castellanii. This pathogen bypasses the elimination mechanism to replicate within amoebae; however, not all amoeba species [...] Read more.

Legionella pneumophila is a facultative intracellular pathogen found in aquatic environments as planktonic cells within biofilms and as intracellular parasites of free-living amoebae such as Acanthamoeba castellanii. This pathogen bypasses the elimination mechanism to replicate within amoebae; however, not all amoeba species support the growth of L. pneumophila. Willaertia magna C2c Maky, a non-pathogenic amoeba, was previously demonstrated to possess the ability to eliminate the L. pneumophila strain Paris. Here, we study the intracellular behaviour of three L. pneumophila strains (Paris, Philadelphia, and Lens) within W. magna C2c Maky and compare this strain to A. castellanii and W. magna Z503, which are used as controls. We observe the intracellular growth of strain Lens within W. magna Z503 and A. castellanii at 22 °C and 37 °C. Strain Paris grows within A. castellanii at any temperature, while it only grows at 22 °C within W. magna Z503. Strain Philadelphia proliferates only within A. castellanii at 37 °C. Within W. magna C2c Maky, none of the three legionella strains exhibit intracellular growth. Additionally, the ability of W. magna C2c Maky to decrease the number of internalized L. pneumophila is confirmed. These results support the idea that W. magna C2c Maky possesses unique behaviour in regard to L. pneumophila strains. Full article

► Show Figures

Figure 1

2019

Jump to: 2024, 2022, 2021, 2020

11 pages, 1253 KiB

Open AccessArticle

Water Quality as a Predictor of Legionella Positivity of Building Water Systems

by David Pierre, Julianne L. Baron, Xiao Ma, Frank P. Sidari III, Marilyn M. Wagener and Janet E. Stout

Pathogens 2019, 8(4), 295; https://doi.org/10.3390/pathogens8040295 - 13 Dec 2019

Cited by 33 | Viewed by 5256

Abstract

Testing drinking water systems for the presence of Legionella colonization is a proactive approach to assess and reduce the risk of Legionnaires’ disease. Previous studies suggest that there may be a link between Legionella positivity in the hot water return line or certain [...] Read more.

Testing drinking water systems for the presence of Legionella colonization is a proactive approach to assess and reduce the risk of Legionnaires’ disease. Previous studies suggest that there may be a link between Legionella positivity in the hot water return line or certain water quality parameters (temperature, free chlorine residual, etc.) with distal site Legionella positivity. It has been suggested that these measurements could be used as a surrogate for testing for Legionella in building water systems. We evaluated the relationship between hot water return line Legionella positivity and other water quality parameters and Legionella colonization in premise plumbing systems by testing 269 samples from domestic cold and hot water samples in 28 buildings. The hot water return line Legionella positivity and distal site positivity only demonstrated a 77.8% concordance rate. Hot water return line Legionella positivity compared to distal site positivity had a sensitivity of 55% and a specificity of 96%. There was poor correlation and a low positive predictive value between the hot water return line and distal outlet positivity. There was no correlation between Legionella distal site positivity and total bacteria (heterotrophic plate count), pH, free chlorine, calcium, magnesium, zinc, manganese, copper, temperature, total organic carbon, or incoming cold-water chlorine concentration. These findings suggest that hot water return line Legionella positivity and other water quality parameters are not predictive of distal site positivity and should not be used alone to determine the building’s Legionella colonization rate and effectiveness of water management programs. Full article

► Show Figures

Figure 1

21 pages, 794 KiB

Open AccessArticle

Advances in Legionella Control by a New Formulation of Hydrogen Peroxide and Silver Salts in a Hospital Hot Water Network

by Luna Girolamini, Ada Dormi, Tiziana Pellati, Paolo Somaroli, Davide Montanari, Andrea Costa, Francesca Savelli, Andrea Martelli, Antonella Grottola, Giulia Fregni Serpini and Sandra Cristino

Pathogens 2019, 8(4), 209; https://doi.org/10.3390/pathogens8040209 - 29 Oct 2019

Cited by 18 | Viewed by 4255

Abstract

Legionella surveillance is an important issue in public health, linked to the severity of disease and the difficulty associated with eradicating this bacterium from the water environment. Different treatments are suggested to reduce Legionella risk, however long-term studies of their efficiency are lacking. [...] Read more.

Legionella surveillance is an important issue in public health, linked to the severity of disease and the difficulty associated with eradicating this bacterium from the water environment. Different treatments are suggested to reduce Legionella risk, however long-term studies of their efficiency are lacking. This study focused on the activity of a new formulation of hydrogen peroxide and silver salts, WTP828, in the hospital hot water network (HWN) to contain Legionella contamination during two years of treatment. The effectiveness of WTP828 was tested measuring physical-chemical and microbiological parameters such as Legionella, Pseudomonas aeruginosa (P. aeruginosa), and a heterotopic plate count (HPC) at 36 °C. Legionella isolates were identified by serotyping and genotyping. WTP 828 induced a reduction in Legionella–positive sites (60% to 36%) and contamination levels (2.12 to 1.7 log₁₀ CFU/L), with isolates belonging to L. pneumophila SG1 (ST1 and ST104), L. anisa and L. rubrilucens widely distributed in HWN. No relevant contamination was found for other parameters tested. The long-term effect of WTP828 on Legionella containment suggest the easy and safe application of this disinfectant, that combined with knowledge of building characteristics, an adequate environmental monitoring and risk assessment plan, become the key elements in preventing Legionella contamination and exposure. Full article

► Show Figures

Graphical abstract

Journal Menu

Journal Browser

Waterborne Pathogens: Epidemiology, Surveillance, Control and Risk

Share This Topical Collection

Editors

Topical Collection Information

Keywords

Related Special Issue

Published Papers (23 papers)

2024

Jump to: 2022, 2021, 2020, 2019

2022

Jump to: 2024, 2021, 2020, 2019

2021

Jump to: 2024, 2022, 2020, 2019

2020

Jump to: 2024, 2022, 2021, 2019

2019

Jump to: 2024, 2022, 2021, 2020

Further Information

Guidelines

MDPI Initiatives

Follow MDPI