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Article ; Online: Wilderness Medical Society Clinical Practice Guidelines on Water Treatment for Wilderness, International Travel, and Austere Situations: 2024 Update.

Backer, Howard D / Derlet, Robert W / Hill, Vincent R

2023 Volume 35, Issue 1_suppl, Page(s) 45S–66S

Abstract: To provide guidance to medical providers, wilderness users, and travelers, the Wilderness Medical Society convened an expert panel to develop evidence-based guidelines for treating water in situations where the potability of available water is not ... ...

Abstract	To provide guidance to medical providers, wilderness users, and travelers, the Wilderness Medical Society convened an expert panel to develop evidence-based guidelines for treating water in situations where the potability of available water is not assured, including wilderness and international travel, areas impacted by disaster, and other areas without adequate sanitation. The guidelines present the available methods for reducing or eliminating microbiological contamination of water for individuals, groups, or households; evaluation of their effectiveness; and practical considerations. The evidence base includes both laboratory and clinical publications. The panel graded the recommendations based on the quality of supporting evidence and the balance between benefits and risks/burdens according to the criteria published by the American College of Chest Physicians.
MeSH term(s)	Humans ; Disasters ; Societies, Medical ; Wilderness Medicine
Language	English
Publishing date	2023-12-27
Publishing country	United States
Document type	Journal Article ; Practice Guideline
ZDB-ID	1238909-2
ISSN	1545-1534 ; 1080-6032
ISSN (online)	1545-1534
ISSN	1080-6032
DOI	10.1177/10806032231218722
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Article ; Online: Detection of Cryptosporidium Recovered from Large-Volume Water Samples Using Dead-End Ultrafiltration.

Kahler, Amy M / Hill, Vincent R

Methods in molecular biology (Clifton, N.J.)

2019 Volume 2052, Page(s) 23–41

Abstract: The procedure described here provides instructions for detection of Cryptosporidium recovered from large-volume water samples. Water samples are collected by dead-end ultrafiltration in the field and ultrafilters are processed in a laboratory. Microbes ... ...

Abstract	The procedure described here provides instructions for detection of Cryptosporidium recovered from large-volume water samples. Water samples are collected by dead-end ultrafiltration in the field and ultrafilters are processed in a laboratory. Microbes recovered from the filters are further concentrated and subjected to Cryptosporidium isolation or nucleic acid extraction methods for the detection of Cryptosporidium oocysts or Cryptosporidium DNA.
MeSH term(s)	Cryptosporidium/genetics ; Cryptosporidium/isolation & purification ; DNA, Protozoan/genetics ; DNA, Protozoan/isolation & purification ; Fluorescent Antibody Technique/methods ; Microscopy, Fluorescence/instrumentation ; Microscopy, Fluorescence/methods ; Oocysts/immunology ; Real-Time Polymerase Chain Reaction/methods ; Ultrafiltration/instrumentation ; Ultrafiltration/methods ; Water/parasitology ; Workflow
Chemical Substances	DNA, Protozoan ; Water (059QF0KO0R)
Language	English
Publishing date	2019-08-27
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-4939-9748-0_3
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Article ; Online: Detection and identification of Giardia species using real-time PCR and sequencing.

Jothikumar, N / Murphy, Jennifer L / Hill, Vincent R

Journal of microbiological methods

2021 Volume 189, Page(s) 106279

Abstract: We report a specific region of Giardia spp. 18S ribosomal RNA (18S rDNA) that serves as an ideal target for quantitative PCR (qPCR) detection and sequencing to identify Giardia species, including the clinically-relevant G. duodenalis, in clinical and ... ...

Abstract	We report a specific region of Giardia spp. 18S ribosomal RNA (18S rDNA) that serves as an ideal target for quantitative PCR (qPCR) detection and sequencing to identify Giardia species, including the clinically-relevant G. duodenalis, in clinical and environmental samples. The presence of multiple copies of the 18S rDNA gene and variations in the selected 18S genomic region enabled the development of a rapid, sensitive qPCR screening method for the detection of Giardia spp. The analytical sensitivity of the Giardia qPCR assay was determined to be a cyst equivalent of 0.4 G. duodenalis cysts per PCR reaction. Amplicon sequencing of the PCR product confirmed Giardia spp. detection and among the 35 sequences obtained, 31, 3 and 1 isolates were classified as belonging to G. duodenalis, G. microti and G. muris, respectively. The TaqMan assay reported here may be useful for the detection of low levels of Giardia in clinical and environmental samples, and further enables the effective use of direct sequencing of the PCR product for Giardia confirmation and to identify major species of Giardia, including G. duodenalis.
MeSH term(s)	DNA, Ribosomal/genetics ; Feces/parasitology ; Genotype ; Giardia/classification ; Giardia/genetics ; Giardiasis/diagnosis ; Giardiasis/parasitology ; RNA, Ribosomal, 18S/genetics ; Real-Time Polymerase Chain Reaction/methods ; Real-Time Polymerase Chain Reaction/standards ; Sequence Analysis, DNA/methods
Chemical Substances	DNA, Ribosomal ; RNA, Ribosomal, 18S
Language	English
Publishing date	2021-07-14
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	604916-3
ISSN	1872-8359 ; 0167-7012
ISSN (online)	1872-8359
ISSN	0167-7012
DOI	10.1016/j.mimet.2021.106279
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Article: Detection and identification of Giardia species using real-time PCR and sequencing

Jothikumar, N. / Murphy, Jennifer L. / Hill, Vincent R.

Journal of microbiological methods. 2021 Oct., v. 189

2021

Abstract	We report a specific region of Giardia spp. 18S ribosomal RNA (18S rDNA) that serves as an ideal target for quantitative PCR (qPCR) detection and sequencing to identify Giardia species, including the clinically-relevant G. duodenalis, in clinical and environmental samples. The presence of multiple copies of the 18S rDNA gene and variations in the selected 18S genomic region enabled the development of a rapid, sensitive qPCR screening method for the detection of Giardia spp. The analytical sensitivity of the Giardia qPCR assay was determined to be a cyst equivalent of 0.4 G. duodenalis cysts per PCR reaction. Amplicon sequencing of the PCR product confirmed Giardia spp. detection and among the 35 sequences obtained, 31, 3 and 1 isolates were classified as belonging to G. duodenalis, G. microti and G. muris, respectively. The TaqMan assay reported here may be useful for the detection of low levels of Giardia in clinical and environmental samples, and further enables the effective use of direct sequencing of the PCR product for Giardia confirmation and to identify major species of Giardia, including G. duodenalis.
Keywords	detection limit ; genes ; genomics ; quantitative polymerase chain reaction ; ribosomal RNA
Language	English
Dates of publication	2021-10
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	604916-3
ISSN	1872-8359 ; 0167-7012
ISSN (online)	1872-8359
ISSN	0167-7012
DOI	10.1016/j.mimet.2021.106279
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Article ; Online: Wilderness Medical Society Clinical Practice Guidelines for Water Disinfection for Wilderness, International Travel, and Austere Situations.

Backer, Howard D / Derlet, Robert W / Hill, Vincent R

Wilderness & environmental medicine

2019 Volume 30, Issue 4S, Page(s) S100–S120

Abstract: To provide guidance to clinicians, the Wilderness Medical Society convened experts to develop evidence-based guidelines for water disinfection in situations where the potability of available water is not ensured, including wilderness and international ... ...

Abstract	To provide guidance to clinicians, the Wilderness Medical Society convened experts to develop evidence-based guidelines for water disinfection in situations where the potability of available water is not ensured, including wilderness and international travel, areas affected by disaster, and other areas without adequate sanitation. The guidelines present the available methods for reducing or eliminating microbiologic contamination of water for individuals, groups, or households; evaluation of their effectiveness; and practical considerations. The evidence evaluation includes both laboratory and clinical publications. The panel graded the recommendations based on the quality of supporting evidence and the balance between benefits and risks or burdens, according to the criteria published by the American College of Chest Physicians.
MeSH term(s)	Disasters ; Disinfection/methods ; Humans ; Practice Patterns, Physicians' ; Societies, Medical ; Travel-Related Illness ; Water Microbiology ; Water Purification/methods ; Wilderness Medicine/methods ; Wilderness Medicine/standards
Language	English
Publishing date	2019-10-23
Publishing country	United States
Document type	Journal Article ; Practice Guideline
ZDB-ID	1238909-2
ISSN	1545-1534 ; 1080-6032
ISSN (online)	1545-1534
ISSN	1080-6032
DOI	10.1016/j.wem.2019.06.006
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Article ; Online: Estimating Waterborne Infectious Disease Burden by Exposure Route, United States, 2014.

Gerdes, Megan E / Miko, Shanna / Kunz, Jasen M / Hannapel, Elizabeth J / Hlavsa, Michele C / Hughes, Michael J / Stuckey, Matthew J / Francois Watkins, Louise K / Cope, Jennifer R / Yoder, Jonathan S / Hill, Vincent R / Collier, Sarah A

Emerging infectious diseases

2023 Volume 29, Issue 7, Page(s) 1357–1366

Abstract: More than 7.15 million cases of domestically acquired infectious waterborne illnesses occurred in the United States in 2014, causing 120,000 hospitalizations and 6,600 deaths. We estimated disease incidence for 17 pathogens according to recreational, ... ...

Abstract	More than 7.15 million cases of domestically acquired infectious waterborne illnesses occurred in the United States in 2014, causing 120,000 hospitalizations and 6,600 deaths. We estimated disease incidence for 17 pathogens according to recreational, drinking, and nonrecreational nondrinking (NRND) water exposure routes by using previously published estimates. In 2014, a total of 5.61 million (95% credible interval [CrI] 2.97-9.00 million) illnesses were linked to recreational water, 1.13 million (95% CrI 255,000-3.54 million) to drinking water, and 407,000 (95% CrI 72,800-1.29 million) to NRND water. Recreational water exposure was responsible for 36%, drinking water for 40%, and NRND water for 24% of hospitalizations from waterborne illnesses. Most direct costs were associated with pathogens found in biofilms. Estimating disease burden by water exposure route helps direct prevention activities. For each exposure route, water management programs are needed to control biofilm-associated pathogen growth; public health programs are needed to prevent biofilm-associated diseases.
MeSH term(s)	Humans ; United States/epidemiology ; Drinking Water ; Communicable Diseases/epidemiology ; Waterborne Diseases/epidemiology ; Water Supply ; Water Microbiology
Chemical Substances	Drinking Water
Language	English
Publishing date	2023-06-21
Publishing country	United States
Document type	Journal Article
ZDB-ID	1380686-5
ISSN	1080-6059 ; 1080-6040
ISSN (online)	1080-6059
ISSN	1080-6040
DOI	10.3201/eid2907.230231
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Article ; Online: A novel photoinduced electron transfer (PET) primer technique for rapid real-time PCR detection of Cryptosporidium spp.

Jothikumar, N / Hill, Vincent R

Biochemical and biophysical research communications

2013 Volume 436, Issue 2, Page(s) 134–139

Abstract: We report the development of a fluorescently labeled oligonucleotide primer that can be used to monitor real-time PCR. The primer has two parts, the 3'-end of the primer is complimentary to the target and a universal 17-mer stem loop at the 5'-end forms ... ...

Abstract	We report the development of a fluorescently labeled oligonucleotide primer that can be used to monitor real-time PCR. The primer has two parts, the 3'-end of the primer is complimentary to the target and a universal 17-mer stem loop at the 5'-end forms a hairpin structure. A fluorescent dye is attached to 5'-end of either the forward or reverse primer. The presence of guanosine residues at the first and second position of the 3' dangling end effectively quenches the fluorescence due to the photo electron transfer (PET) mechanism. During the synthesis of nucleic acid, the hairpin structure is linearized and the fluorescence of the incorporated primer increases several-fold due to release of the fluorescently labeled tail and the absence of guanosine quenching. As amplicons are synthesized during nucleic acid amplification, the fluorescence increase in the reaction mixture can be measured with commercially available real-time PCR instruments. In addition, a melting procedure can be performed to denature the double-stranded amplicons, thereby generating fluorescence peaks that can differentiate primer dimers and other non-specific amplicons if formed during the reaction. We demonstrated the application of PET-PCR for the rapid detection and quantification of Cryptosporidium parvum DNA. Comparison with a previously published TaqMan® assay demonstrated that the two real-time PCR assays exhibited similar sensitivity for a dynamic range of detection of 6000-0.6 oocysts per reaction. PET PCR primers are simple to design and less-expensive than dual-labeled probe PCR methods, and should be of interest for use by laboratories operating in resource-limited environments.
MeSH term(s)	Base Sequence ; Cryptosporidium parvum/genetics ; DNA Primers/chemistry ; DNA Primers/genetics ; DNA, Protozoan/genetics ; Electron Transport/radiation effects ; Fluorescent Dyes/chemistry ; Nucleic Acid Conformation/radiation effects ; Oocysts/metabolism ; Real-Time Polymerase Chain Reaction/methods ; Reproducibility of Results
Chemical Substances	DNA Primers ; DNA, Protozoan ; Fluorescent Dyes
Language	English
Publishing date	2013-06-28
Publishing country	United States
Document type	Journal Article
ZDB-ID	205723-2
ISSN	1090-2104 ; 0006-291X ; 0006-291X
ISSN (online)	1090-2104 ; 0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2013.05.032
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Article: Calculation and uncertainty of zeta potentials of microorganisms in a 1:1 electrolyte with a conductivity similar to surface water

Polaczyk, Amy L / Amburgey, James E / Alansari, Amir / Poler, Jordan C / Propato, Marco / Hill, Vincent R

Colloids and surfaces. 2020 Feb. 05, v. 586

2020

Abstract: The electrophoretic mobilities (EPM’s) of fifteen different microbes (6 viruses, 5 vegetative bacteria, 2 bacterial endospores, 2 protozoa) and one microbial particle surrogate (Polystyrene microspheres) were measured, and five models were used to ... ...

Abstract	The electrophoretic mobilities (EPM’s) of fifteen different microbes (6 viruses, 5 vegetative bacteria, 2 bacterial endospores, 2 protozoa) and one microbial particle surrogate (Polystyrene microspheres) were measured, and five models were used to convert EPM's of these microorganisms to zeta potentials. The Helmholtz-Smoluchowski, Hückel-Onsager, Henry, modified Booth, and O’Brien and Hunter models were compared over their ranges of applicability for various microbes in a weak electrolyte solution intended to simulate the conductivity of surface water. The results from each of the models were compared by assessing the magnitude of the error due to inherent limitations of the models and comparing it to the error associated with the measurement of the EPM. Results indicated that differences imparted to the calculated zeta potentials by double layer distortion corrections were typically smaller than the uncertainty of the EPM measurement from which the zeta potential value was calculated. Based on our analyses, the Helmholtz-Smoluchowski equation was most appropriate for application to bacteria (vegetative and endospores) and parasites, while the Henry or modified Booth models were necessary for viruses. Zeta potential calculations with corresponding uncertainty values are presented for each of the microbes and the surrogate for each of the five models studied. A zone chart was created to help avoid unnecessary error in calculating microbial zeta potentials that can exceed 50%.
Keywords	Protozoa ; bacteria ; electrolytes ; electrophoresis ; endospores ; equations ; microparticles ; models ; parasites ; polystyrenes ; surface water ; uncertainty ; viruses ; zeta potential
Language	English
Dates of publication	2020-0205
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	1500517-3
ISSN	0927-7757
ISSN	0927-7757
DOI	10.1016/j.colsurfa.2019.124097
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Article: A novel photoinduced electron transfer (PET) primer technique for rapid real-time PCR detection of Cryptosporidium spp

Jothikumar, N / Hill, Vincent R

Biochemical and biophysical research communications. 2013 June 28, v. 436, no. 2

2013

Abstract: We report the development of a fluorescently labeled oligonucleotide primer that can be used to monitor real-time PCR. The primer has two parts, the 3′-end of the primer is complimentary to the target and a universal 17-mer stem loop at the 5′-end forms ... ...

Abstract	We report the development of a fluorescently labeled oligonucleotide primer that can be used to monitor real-time PCR. The primer has two parts, the 3′-end of the primer is complimentary to the target and a universal 17-mer stem loop at the 5′-end forms a hairpin structure. A fluorescent dye is attached to 5′-end of either the forward or reverse primer. The presence of guanosine residues at the first and second position of the 3′ dangling end effectively quenches the fluorescence due to the photo electron transfer (PET) mechanism. During the synthesis of nucleic acid, the hairpin structure is linearized and the fluorescence of the incorporated primer increases several-fold due to release of the fluorescently labeled tail and the absence of guanosine quenching. As amplicons are synthesized during nucleic acid amplification, the fluorescence increase in the reaction mixture can be measured with commercially available real-time PCR instruments. In addition, a melting procedure can be performed to denature the double-stranded amplicons, thereby generating fluorescence peaks that can differentiate primer dimers and other non-specific amplicons if formed during the reaction. We demonstrated the application of PET-PCR for the rapid detection and quantification of Cryptosporidium parvum DNA. Comparison with a previously published TaqMan® assay demonstrated that the two real-time PCR assays exhibited similar sensitivity for a dynamic range of detection of 6000–0.6 oocysts per reaction. PET PCR primers are simple to design and less-expensive than dual-labeled probe PCR methods, and should be of interest for use by laboratories operating in resource-limited environments.
Keywords	Cryptosporidium parvum ; DNA ; DNA primers ; electron transfer ; equipment ; fluorescence ; fluorescent dyes ; guanosine ; melting ; oocysts ; quantitative polymerase chain reaction ; rapid methods
Language	English
Dates of publication	2013-0628
Size	p. 134-139.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	205723-2
ISSN	0006-291X ; 0006-291X
ISSN (online)	0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2013.05.032
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Article ; Online: Recovery of diverse microbes in high turbidity surface water samples using dead-end ultrafiltration.

Mull, Bonnie / Hill, Vincent R

Journal of microbiological methods

2012 Volume 91, Issue 3, Page(s) 429–433

Abstract: Dead-end ultrafiltration (DEUF) has been reported to be a simple, field-deployable technique for recovering bacteria, viruses, and parasites from large-volume water samples for water quality testing and waterborne disease investigations. While DEUF has ... ...

Abstract	Dead-end ultrafiltration (DEUF) has been reported to be a simple, field-deployable technique for recovering bacteria, viruses, and parasites from large-volume water samples for water quality testing and waterborne disease investigations. While DEUF has been reported for application to water samples having relatively low turbidity, little information is available regarding recovery efficiencies for this technique when applied to sampling turbid water samples such as those commonly found in lakes and rivers. This study evaluated the effectiveness of a DEUF technique for recovering MS2 bacteriophage, enterococci, Escherichia coli, Clostridium perfringens, and Cryptosporidium parvum oocysts in surface water samples having elevated turbidity. Average recovery efficiencies for each study microbe across all turbidity ranges were: MS2 (66%), C. parvum (49%), enterococci (85%), E. coli (81%), and C. perfringens (63%). The recovery efficiencies for MS2 and C. perfringens exhibited an inversely proportional relationship with turbidity, however no significant differences in recovery were observed for C. parvum, enterococci, or E. coli. Although ultrafilter clogging was observed, the DEUF method was able to process 100-L surface water samples at each turbidity level within 60 min. This study supports the use of the DEUF method for recovering a wide array of microbes in large-volume surface water samples having medium to high turbidity.
MeSH term(s)	Animals ; Bacteria/classification ; Bacteria/isolation & purification ; Biodiversity ; Fresh Water/analysis ; Fresh Water/microbiology ; Fresh Water/parasitology ; Parasites/chemistry ; Parasites/isolation & purification ; Ultrafiltration/methods ; Viruses/classification ; Viruses/isolation & purification ; Water Microbiology
Language	English
Publishing date	2012-10-12
Publishing country	Netherlands
Document type	Evaluation Study ; Journal Article ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	604916-3
ISSN	1872-8359 ; 0167-7012
ISSN (online)	1872-8359
ISSN	0167-7012
DOI	10.1016/j.mimet.2012.10.001
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