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Food and Water-Borne Pathogens: Epidemiology, Genomics, Virulence and Resistance Profiles

A special issue of Microorganisms (ISSN 2076-2607). This special issue belongs to the section "Environmental Microbiology".

Deadline for manuscript submissions: 30 June 2024 | Viewed by 5354

Special Issue Editors


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Guest Editor
Instituto Nacional de Saúde Dr. Ricardo Jorge, Lisbon, Portugal
Interests: enterobacteria; foodborne pathogens; molecular epidemiology; AMR, virulence; molecular microbiology; one health; WGS

E-Mail Website
Guest Editor Assistant
Instituto Nacional de Saúde Dr. Ricardo Jorgedisabled, Lisbon, Portugal
Interests: foodborne pathogens

Special Issue Information

Dear Colleagues,

We are pleased to invite you to submit your original research manuscripts to this Special Issue.

Food- and water-borne infections cause a major economic burden, having a great impact on food safety, human and animal health worldwide. These infections are widely spread through the consumption of food and/or water contaminated by bacteria or their toxins, parasites, and viruses.

This Special Issue aims to better understand the epidemiology, ecology, genomics, virulence, and transmission mechanisms of resistance markers of some of the most important food- and waterborne pathogens, covering the following topics:

  • Food- and waterborne bacterial pathogens (Campylobacter, Salmonella, Yersinia, coli, L.monocytogenes, Shigella, Vibrio, S. aureus, among others);
  • Food- and waterborne viral pathogens (Hepatitis A, Norovirus, among others);
  • Food- and waterborne parasites (gondii, T.spiralis, Criptosporidium, among others);
  • Virulence factors and mechanisms of infection;
  • Antimicrobial resistance;
  • Mobile genetic elements;
  • Persistence and biofilms;
  • Whole-genome sequencing;
  • Public health and epidemiology;
  • Trends of microbial pathogens;
  • Influence of environmental changes in gene expression;
  • Animal health;
  • Food safety;
  • Water quality;
  • Prevention and control of food- and waterborne pathogens.

Dr. Angela Pista
Dr. Rita Batista
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Microorganisms is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • antimicrobial resistance
  • mobile genetic elements
  • persistence and biofilms
  • whole-genome sequencing
  • public health and epidemiology
  • trends of microbial pathogens
  • influence of environmental changes in gene expression
  • animal health
  • food safety
  • water quality
  • Prevention and control of food- and waterborne pathogens.

Published Papers (4 papers)

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Research

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13 pages, 5851 KiB  
Article
Complete Genome Sequence and Pan-Genome Analysis of Shewanella oncorhynchi Z-P2, a Siderophore Putrebactin-Producing Bacterium
by Ying Zhang, Mengjie Pan, Qiaoyun Wang, Lan Wang and Li Liao
Microorganisms 2023, 11(12), 2961; https://doi.org/10.3390/microorganisms11122961 - 11 Dec 2023
Viewed by 1274
Abstract
In this study, we reported the complete genome sequence of Shewanella oncorhynchi for the first time. S. oncorhynchi Z-P2 is a bacterium that produces the siderophore putrebactin. Its genome consists of a circular chromosome of 5,034,612 bp with a G + C content [...] Read more.
In this study, we reported the complete genome sequence of Shewanella oncorhynchi for the first time. S. oncorhynchi Z-P2 is a bacterium that produces the siderophore putrebactin. Its genome consists of a circular chromosome of 5,034,612 bp with a G + C content of 45.4%. A total of 4544 protein-coding genes, 109 tRNAs and 31 rRNAs were annotated by the RAST. Five non-ribosomal peptide synthetase (NRPS) and polyketide synthetase (PKS) gene clusters were identified by the antiSMASH analysis. The pan-genome analysis of Z-P2 and 10 Shewanella putrefaciens revealed 9228 pan-gene clusters and 2681 core gene clusters, with Z-P2 having 618 unique gene clusters. Additionally, the gene cluster involved in putrebactin biosynthesis in Z-P2 was annotated, and the mechanism of putrebactin biosynthesis was analyzed. The putrebactin produced by Z-P2 was detected using UPLC-MS analysis, with an [M + H]+ molecular ion at m/z 373.21. These findings provide valuable support for further research on the genetic engineering of putrebactin biosynthetic genes of Z-P2 and their potential applications. Full article
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Circular genome map of <span class="html-italic">S. oncorhynchi</span> Z-P2. From outer to the inner: ring 1 represents genome size; ring 2 represents forward and reverse chains of coding DNA sequences (CDSs) colored according to COG categories; ring 3 represents forward and reverse chains of non-coding RNA (ncRNA) genes; ring 4 represents the G + C content (red, higher than the average; blue, lower than the average); ring 5 represents the GC skew (orange, positive skew; green, negative skew). Numbers in parentheses represent the count of genes with this functional type of COG.</p>
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<p>RAST subsystem distribution of <span class="html-italic">S. oncorhynchi</span> Z-P2.</p>
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<p>KEGG pathways annotation of <span class="html-italic">S. oncorhynchi</span> Z-P2. The level 1 pathways (in black) and level 2 pathways (in different color according to level 1 categories) were described on the left.</p>
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<p>Neighbor-joining tree with a bootstrap test (1000 replicates) based on the 16S rDNA gene sequences of <span class="html-italic">S. oncorhynchi</span> Z-P2. The scale bar indicates the evolutionary distance in nucleotide substitutions per site.</p>
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<p>The secondary metabolite biosynthetic gene clusters identified in <span class="html-italic">S. oncorhynchi</span> Z-P2. (<b>A</b>): APE biosynthetic gene cluster; (<b>B</b>): beta-lactone biosynthetic gene cluster; (<b>C</b>): putrebactin biosynthetic gene cluster; (<b>D</b>): eicosapentaenoic acid biosynthetic gene cluster; (<b>E</b>): RiPP biosynthetic gene cluster.</p>
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<p>Pan-genome analysis of <span class="html-italic">S. oncorhynchi</span> Z-P2 and 10 <span class="html-italic">S. putrefaciens</span> strains. (<b>A</b>) The number of pan-gene clusters (blue) and core gene clusters (orange) among Z-P2 and different <span class="html-italic">S. putrefaciens</span> strains. (<b>B</b>) Phylogenetic tree of Z-P2 and 10 <span class="html-italic">S. putrefaciens</span> strains based on whole genomes. (<b>C</b>) Heatmap and hierarchical clustering based on pairwise average nucleotide identity (ANI) values.</p>
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<p>Distribution of genes among <span class="html-italic">S. oncorhynchi</span> Z-P2 and 10 <span class="html-italic">S. putrefaciens</span> strains. (<b>A</b>) Upset plot of comparisons among unique genes of Z-P2 and <span class="html-italic">S. putrefaciens</span> strains. (<b>B</b>) Pan-genome profile of Z-P2 and <span class="html-italic">S. putrefaciens</span> strains. COG annotation showing the core genes and unique genes of Z-P2 and <span class="html-italic">S. putrefaciens</span> strains.</p>
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<p>Putrebactin biosynthesis gene cluster (<b>A</b>) and proposed biosynthetic pathway (<b>B</b>) of <span class="html-italic">S. oncorhynchi</span> Z-P2.</p>
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<p>Siderophore production of <span class="html-italic">S. oncorhynchi</span> Z-P2. <b>Left</b>: CAS solution and uninoculated broth. <b>Right</b>: CAS solution and cell-free supernatant of Z-P2.</p>
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<p>Putrebactin production of <span class="html-italic">S. oncorhynchi</span> Z-P2. (<b>A</b>) Base peak chromatogram (BPC). (<b>B</b>) Mass spectrum.</p>
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12 pages, 1243 KiB  
Article
Lethality Validation for Human Pathogenic Salmonella enterica on Chicken Feathers and Blood during Simulated Commercial Low-Temperature Dry Rendering
by Aime L. Shimwa Mvuyekure, Rosana G. Moreira and Thomas Matthew Taylor
Microorganisms 2023, 11(8), 2071; https://doi.org/10.3390/microorganisms11082071 - 12 Aug 2023
Viewed by 922
Abstract
Poultry rendering is the process of upcycling inedible poultry carcass materials into useful animal food/feed components as well as other valuable commercial products. Microbiological safety validation is nonetheless critical to ensuring the prevention of food safety hazard(s) transmission. This study determined the death [...] Read more.
Poultry rendering is the process of upcycling inedible poultry carcass materials into useful animal food/feed components as well as other valuable commercial products. Microbiological safety validation is nonetheless critical to ensuring the prevention of food safety hazard(s) transmission. This study determined the death kinetics of the thermotolerant Salmonella enterica serovar Senftenberg isolate 775W in chicken feathers and blood in low-temperature dry rendering (i.e., no direct contact with heating medium) to validate pathogen inactivation in commercial processing. Chicken feathers and blood were inoculated with Salmonella Senftenberg 775W and heated to 60, 70, or 80 °C for up to 60, 20, and 5 min, respectively. Three identically completed replicates (N = 3) for each product were conducted. Pathogen inactivation data were fitted to a non-linear model, providing for the detection and characterization of shoulder, log-linear death, and tailing components in death curves. The analysis showed a >7-log10 reduction in Salmonella was achieved across all processing temperatures, with t7D values (time for 7.0 log-cycle lethality) ranging from 21.68, 7.30, and 4.26 min for feathers and 18.38, 5.03, and 2.79 min in blood at 60, 70, and 80 °C, respectively. Study findings validate that low-temperature processing conditions can inactivate Salmonella in poultry-rendered offal. Full article
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Figure 1

Figure 1
<p>Growth of <span class="html-italic">Salmonella</span> Senftenberg 775W in tryptic soy broth at 37 °C. Symbols depict least squares means from three replications (<span class="html-italic">N</span> = 3); error bars depict the standard deviation of the mean.</p>
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<p>Survival of rifampicin-resistant <span class="html-italic">Salmonella</span> Senftenberg 775W inoculated on chicken feathers predicted by Geeraerd et al.’s [<a href="#B17-microorganisms-11-02071" class="html-bibr">17</a>] model at temperatures 60, 70, and 80 °C. Symbols depict the means of three identical complete replicates (<span class="html-italic">N</span> = 3); error bars depict one sample standard deviation from the mean. Surviving <span class="html-italic">Salmonella</span> were enumerated on tryptic soy agar supplemented with 100 μg/mL rifampicin following 24 h of incubation at 37 °C.</p>
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<p>Survival of rifampicin-resistant <span class="html-italic">Salmonella</span> Senftenberg 775W inoculated on chicken blood predicted by Geeraerd et al.’s [<a href="#B17-microorganisms-11-02071" class="html-bibr">17</a>] model at temperatures 60, 70, and 80 °C. Symbols depict the means of three identical complete replicates (<span class="html-italic">N</span> = 3); error bars depict one sample standard deviation from the mean. Surviving <span class="html-italic">Salmonella</span> were enumerated on tryptic soy agar supplemented with 100 μg/mL rifampicin following 24 h of incubation at 37 °C.</p>
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11 pages, 2831 KiB  
Article
Detection of Cyclomodulin CNF-1 Toxin-Producing Strains of Escherichia coli in Pig Kidneys at a Slaughterhouse
by Arturo Herrera-Vázquez, Rebeca Arellano-Aranda, Daniel Hernández-Cueto, Esmeralda Rodríguez-Miranda, Sergio López-Briones and Marco Antonio Hernández-Luna
Microorganisms 2023, 11(8), 2065; https://doi.org/10.3390/microorganisms11082065 - 11 Aug 2023
Cited by 1 | Viewed by 979
Abstract
Food is often contaminated with Escherichia coli (E. coli) bacteria strains, which have been associated with different diseases, including urinary tract infections. The consumption of meat by humans is a potential route of transmission of antimicrobial resistance, and food-producing animals have been associated [...] Read more.
Food is often contaminated with Escherichia coli (E. coli) bacteria strains, which have been associated with different diseases, including urinary tract infections. The consumption of meat by humans is a potential route of transmission of antimicrobial resistance, and food-producing animals have been associated as a major reservoir of resistant bacterial strains. The aim of this study was to determine the presence of the E. coli strains producing the CNF-1 toxin in pig kidneys. Pig kidneys were collected from a Mexican slaughterhouse and classified according to their coloration into reddish kidneys (RK) and yellowish kidneys (YK). A tissue sample from each kidney was processed for histological analysis, the presence of E. coli was determined by conventional PCR assay, and the CNF-1 toxin was detected by both conventional PCR and Western blotting. Herein, an inflammatory cell infiltrate was found in all collected kidneys, regardless of macroscopic differences. Surprisingly, E. coli and the CNF-1 toxin were detected in all kidney samples. We clearly demonstrate contamination by CNF-1 toxin-producing E. coli in pork kidneys from a slaughterhouse, even in those without apparent damage. This suggests that pork may serve as a reservoir for pathogens, representing an important risk to human health. Full article
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Figure 1

Figure 1
<p>Macroscopic Characteristics of Pig Kidneys. Pig kidneys were sagittally dissected. Reddish kidneys (RK) without apparent damage and yellowish kidneys with possible damage (YK) are shown in (<b>A</b>,<b>B</b>), respectively.</p>
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<p>Cellular infiltration in pig kidney tissues. A slice of renal cortex was evaluated histologically by hematoxylin and eosin staining and analyzed by conventional light microscopy at 20× and 40× magnification, respectively. Cellular infiltrate was observed in both reddish kidney (<b>A</b>,<b>B</b>) and yellowish kidney tissue (<b>C</b>,<b>D</b>). The arrows indicate the cellular infiltrate in each sample.</p>
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<p>Quantitative assessment of inflammatory cell infiltrate. Cellular infiltration was analyzed by conventional light microscopy, counting 10 different fields of each kidney sample. The bars represent the average of the infiltrating inflammatory cell counts in at least 5 different RK (black bar) and YK (gray bar) samples, respectively. * <span class="html-italic">p</span> &lt; 0.05 by U-Mann–Whitney test.</p>
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<p>Detection of bacterial ribosome 16S subunit and <span class="html-italic">E. coli</span> identification. Total DNA was purified from renal cortex tissue. Amplification of 16S ribosomal subunit and <span class="html-italic">E. coli</span> specific gene was performed by end-point PCR. Amplification of the 16S ribosome subunit (<b>A</b>,<b>B</b>) and the <span class="html-italic">E. coli</span>-specific gene (<b>C</b>,<b>D</b>) is shown in six different samples of reddish kidneys (RK 1–6) and six different samples of yellowish kidneys (YK1–6), respectively. As a positive control for both 16S ribosome subunit and <span class="html-italic">E. coli</span>, a mouse intestinal microbiota DNA sample was used.</p>
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<p>Gene amplification of <span class="html-italic">cnf-1</span>. Total DNA was purified from renal cortex tissue. Gene amplification of <span class="html-italic">cnf-1</span> was performed by endpoint PCR. The <span class="html-italic">cnf-1</span> gen is shown in four different samples of reddish kidneys (RK 1–4) and four different samples of yellowish kidneys (YK1–4), respectively. Amplification of the 16S ribosomal subunit was used as a control.</p>
Full article ">Figure 6
<p>Detection of CNF-1 protein expression levels. Total protein was purified from renal cortex and subjected to SDS-PAGE. CNF-1 toxin protein was determined by Western blot assay. The protein expression levels of CNF-1 toxin are shown in three different reddish kidney samples (RK 1–3) and three different yellowish kidney samples (YK1–3), respectively. Detection of β-actin was used as a control to compare CNF-1 toxin expression levels.</p>
Full article ">

Review

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58 pages, 5337 KiB  
Review
Opportunistic Pathogens in Drinking Water Distribution Systems—A Review
by Mark W. LeChevallier, Toby Prosser and Melita Stevens
Microorganisms 2024, 12(5), 916; https://doi.org/10.3390/microorganisms12050916 - 30 Apr 2024
Viewed by 1479
Abstract
In contrast to “frank” pathogens, like Salmonella entrocolitica, Shigella dysenteriae, and Vibrio cholerae, that always have a probability of disease, “opportunistic” pathogens are organisms that cause an infectious disease in a host with a weakened immune system and rarely in [...] Read more.
In contrast to “frank” pathogens, like Salmonella entrocolitica, Shigella dysenteriae, and Vibrio cholerae, that always have a probability of disease, “opportunistic” pathogens are organisms that cause an infectious disease in a host with a weakened immune system and rarely in a healthy host. Historically, drinking water treatment has focused on control of frank pathogens, particularly those from human or animal sources (like Giardia lamblia, Cryptosporidium parvum, or Hepatitis A virus), but in recent years outbreaks from drinking water have increasingly been due to opportunistic pathogens. Characteristics of opportunistic pathogens that make them problematic for water treatment include: (1) they are normally present in aquatic environments, (2) they grow in biofilms that protect the bacteria from disinfectants, and (3) under appropriate conditions in drinking water systems (e.g., warm water, stagnation, low disinfectant levels, etc.), these bacteria can amplify to levels that can pose a public health risk. The three most common opportunistic pathogens in drinking water systems are Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa. This report focuses on these organisms to provide information on their public health risk, occurrence in drinking water systems, susceptibility to various disinfectants, and other operational practices (like flushing and cleaning of pipes and storage tanks). In addition, information is provided on a group of nine other opportunistic pathogens that are less commonly found in drinking water systems, including Aeromonas hydrophila, Klebsiella pneumoniae, Serratia marcescens, Burkholderia pseudomallei, Acinetobacter baumannii, Stenotrophomonas maltophilia, Arcobacter butzleri, and several free-living amoebae including Naegleria fowleri and species of Acanthamoeba. The public health risk for these microbes in drinking water is still unclear, but in most cases, efforts to manage Legionella, mycobacteria, and Pseudomonas risks will also be effective for these other opportunistic pathogens. The approach to managing opportunistic pathogens in drinking water supplies focuses on controlling the growth of these organisms. Many of these microbes are normal inhabitants in biofilms in water, so the attention is less on eliminating these organisms from entering the system and more on managing their occurrence and concentrations in the pipe network. With anticipated warming trends associated with climate change, the factors that drive the growth of opportunistic pathogens in drinking water systems will likely increase. It is important, therefore, to evaluate treatment barriers and management activities for control of opportunistic pathogen risks. Controls for primary treatment, particularly for turbidity management and disinfection, should be reviewed to ensure adequacy for opportunistic pathogen control. However, the major focus for the utility’s opportunistic pathogen risk reduction plan is the management of biological activity and biofilms in the distribution system. Factors that influence the growth of microbes (primarily in biofilms) in the distribution system include, temperature, disinfectant type and concentration, nutrient levels (measured as AOC or BDOC), stagnation, flushing of pipes and cleaning of storage tank sediments, and corrosion control. Pressure management and distribution system integrity are also important to the microbial quality of water but are related more to the intrusion of contaminants into the distribution system rather than directly related to microbial growth. Summarizing the identified risk from drinking water, the availability and quality of disinfection data for treatment, and guidelines or standards for control showed that adequate information is best available for management of L. pneumophila. For L. pneumophila, the risk for this organism has been clearly established from drinking water, cases have increased worldwide, and it is one of the most identified causes of drinking water outbreaks. Water management best practices (e.g., maintenance of a disinfectant residual throughout the distribution system, flushing and cleaning of sediments in pipelines and storage tanks, among others) have been shown to be effective for control of L. pneumophila in water supplies. In addition, there are well documented management guidelines available for the control of the organism in drinking water distribution systems. By comparison, management of risks for Mycobacteria from water are less clear than for L. pneumophila. Treatment of M. avium is difficult due to its resistance to disinfection, the tendency to form clumps, and attachment to surfaces in biofilms. Additionally, there are no guidelines for management of M. avium in drinking water, and one risk assessment study suggested a low risk of infection. The role of tap water in the transmission of the other opportunistic pathogens is less clear and, in many cases, actions to manage L. pneumophila (e.g., maintenance of a disinfectant residual, flushing, cleaning of storage tanks, etc.) will also be beneficial in helping to manage these organisms as well. Full article
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Figure 1
<p>Etiology of drinking water associated outbreaks (<span class="html-italic">n</span> = 928) by year, US, 1971–2014. Source: Benedict et al. [<a href="#B9-microorganisms-12-00916" class="html-bibr">9</a>].</p>
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<p>Examples of biofilms in water systems. From: LeChevallier [<a href="#B33-microorganisms-12-00916" class="html-bibr">33</a>].</p>
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<p>Average and standard deviation of water temperatures in treated water storages within a distribution system in Australia. Data from 2013–2022.</p>
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<p>Percentage of chlorine residuals greater than 0.2 mg/L in three regions of an Australian water system.</p>
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<p>AOC levels in 94 North American water systems. From Volk and LeChevallier [<a href="#B29-microorganisms-12-00916" class="html-bibr">29</a>].</p>
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<p>BDOC levels in 30 North American water systems. From Volk and LeChevallier [<a href="#B29-microorganisms-12-00916" class="html-bibr">29</a>].</p>
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<p>Decision tree for coliform occurrences in drinking water. Abbreviations: T °C, temperature in degrees Celsius, AOC, assimilable organic carbon. From Volk and LeChevallier [<a href="#B29-microorganisms-12-00916" class="html-bibr">29</a>].</p>
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<p>Legionella cases in Victoria, 2012 to 2020.</p>
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<p>Comparison of Legionnaires’ Disease (LD) and Pontiac Fever (PF) Cases 2006–2017. Adapted from Hamilton et al. [<a href="#B120-microorganisms-12-00916" class="html-bibr">120</a>].</p>
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<p>Concentration of culturable Legionella during outbreaks (red-orange) and routine monitoring (green). The black line is the 5 × 10<sup>4</sup> cfu/L action level as a break between sporadic cases and outbreaks. From NASEM [<a href="#B111-microorganisms-12-00916" class="html-bibr">111</a>].</p>
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<p>Relationship between <span class="html-italic">L. pneumophila</span> concentration and free chlorine residual. From LeChevallier [<a href="#B34-microorganisms-12-00916" class="html-bibr">34</a>].</p>
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<p>Relationship between temperature and concentration of <span class="html-italic">L. pneumophila</span>. From LeChevallier [<a href="#B34-microorganisms-12-00916" class="html-bibr">34</a>].</p>
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<p>Impact of distribution system flushing at site 824 to control the occurrence of culturable <span class="html-italic">L. pneumophila</span>. Symbols: d, downstream f site 824; u, upstream of site 824. From LeChevallier [<a href="#B34-microorganisms-12-00916" class="html-bibr">34</a>].</p>
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<p>Projected NTM cases in Queensland, Australia from 2020 to 2040. From Ratnatunga et al. [<a href="#B194-microorganisms-12-00916" class="html-bibr">194</a>].</p>
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<p>Occurrence of <span class="html-italic">M. avium</span> and <span class="html-italic">M. intracellulare</span> in distribution system biofilm samples (N = 55). Site # - site number. From Falkinham et al. [<a href="#B50-microorganisms-12-00916" class="html-bibr">50</a>].</p>
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<p>Disinfection of mycobacteria by free chlorine. Experimental conditions: pH 7.0, 25 °C, initial free chlorine concentration 0.5 mg/L.Adapted from Le Dantec et al. [<a href="#B216-microorganisms-12-00916" class="html-bibr">216</a>].</p>
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<p>Impact of pipe composition on biofilm growth of <span class="html-italic">M. avium</span>. Adapted from LeChevallier et al. [<a href="#B231-microorganisms-12-00916" class="html-bibr">231</a>].</p>
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<p>Seasonal occurrence of P. aeruginosa in Croatia. Adapted from Vukić Lušić et al. [<a href="#B246-microorganisms-12-00916" class="html-bibr">246</a>].</p>
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