Detection of Bacterial Indicators and Human and Bovine Enteric Viruses in Surface Water and Groundwater Sources Potentially Impacted by Animal and Human Wastes in Lower Yakima Valley, Washington

Gibson, Kristen E.; Schwab, Kellogg J.

doi:10.1128/aem.01407-10

Cited by 42 publications

(28 citation statements)

References 26 publications

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“…Swine pathogens were indicated by stx 2e , STII, PAdV, and PTV (Table 1). Genogroup II (GII) norovirus was detected in 2 of 3 slurry samples by using HuNoV GII-specific primers and probe (26) and was confirmed as swine norovirus by sequencing per reference 27. The STII gene was quantifiable by qPCR (7.8 log 10 gene copies/100 g [wet weight]) in swine manure slurry from one of the three farms.…”

Section: Resultsmentioning

confidence: 99%

“…Large-volume water samples (n ϭ 5; 40 to 100 liters) were concentrated by tangential-flow ultrafiltration (UF) as described previously (26). Prior to nucleic acid extraction, 1 ml of each swine manure slurry sample was clarified by centrifugation at 3,000 ϫ g for 5 min at 4°C.…”

Section: 6-25%mentioning

confidence: 99%

“…Extracted NA was aliquoted and archived at Ϫ80°C until analysis. Amplification of viral DNA and RNA targets was performed as described previously for human norovirus (HuNoV), bovine enterovirus (BoEV), bovine norovirus (BoNoV), human adenovirus (HuAdV), and human polyomavirus (HuPyV) (26). Information on hepatitis E virus (HEV), pan-enterovirus (pan-EV), porcine teschovirus (PTV), and porcine adenovirus (PAdV) assays is in Table 3.…”

Section: 6-25%mentioning

confidence: 99%

“…Aliquots (200 l) of the secondary concentrates and pellets were processed for total NA extraction using the QIAamp MinElute virus spin kit (Qiagen) as described previously (26). Extracted NA was aliquoted and archived at Ϫ80°C until analysis.…”

Section: 6-25%mentioning

confidence: 99%

“…All assays were validated with positive and negative (nontarget NA and DEPC-treated water) controls. An internal standard assay for the identification of inhibition in qPCR and RT-qPCR was applied to each NA extract prior to analysis for target viruses (26). RT-qPCR assay conditions for five of the assays (HuNoV GI and GII, pan-EV, BoEV, BoNoV, and PTV) were RT for 30 min at 50°C, denaturation for 15 min at 95°C, 40 cycles at 95°C for 15s, and annealing/extension at 60°C (57°C for BoNoV) for 60 s. qPCR conditions for HuAdV were 15 min at 95°C, 40 cycles at 94°C for 15 s, and annealing/extension at 60°C for 60 s; qPCR conditions for HuPyV were 15 min at 95°C, 40 cycles at 95°C for 15 s and 55°C for 15 s, and extension at 60°C for 60 s; and qPCR conditions for PAdV were 15 min at 95°C, 40 cycles at 95°C for 10 s and 55°C for 20 s, and extension at 60°C for 20 s. All qPCR analyses were performed utilizing a positive control for each target and DEPC-treated water as the negative control with each run.…”

Section: 6-25%mentioning

confidence: 99%

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Genes Indicative of Zoonotic and Swine Pathogens Are Persistent in Stream Water and Sediment following a Swine Manure Spill

Haack

Duris

Kolpin

et al. 2015

Appl Environ Microbiol

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Manure spills into streams are relatively frequent, but no studies have characterized stream contamination with zoonotic and veterinary pathogens, or fecal chemicals, following a spill. We tested stream water and sediment over 25 days and downstream for 7.6 km for the following: fecal indicator bacteria (FIB), the fecal indicator chemicals cholesterol and coprostanol, 20 genes for zoonotic and swine-specific bacterial pathogens by presence/absence PCR for viable cells, one swine-specific Escherichia coli toxin gene (STII gene) by quantitative PCR (qPCR), and nine human and animal viruses by qPCR or reverse transcription-qPCR. Twelve days postspill, and 4.2 km downstream, water concentrations of FIB, cholesterol, and coprostanol were 1 to 2 orders of magnitude greater than those detected before, or above, the spill, and genes indicating viable zoonotic or swine-infectious Escherichia coli were detected in water or sediment. STII gene levels increased from undetectable before or above the spill to 10 5 copies/100 ml of water 12 days postspill. Thirteen of 14 water (8/9 sediment) samples had viable STII-carrying cells postspill. Eighteen days postspill, porcine adenovirus and teschovirus were detected 5.6 km downstream. FIB concentrations (per gram [wet weight]) in sediment were greater than in water, and sediment was a continuous reservoir of genes and chemicals postspill. Constituent concentrations were much lower, and detections less frequent, in a runoff event (200 days postspill) following manure application, although the swine-associated STII and stx 2e genes were detected. Manure spills are an underappreciated pathway for livestock-derived contaminants to enter streams, with persistent environmental outcomes and the potential for human and veterinary health consequences.A cross the United States, animal agriculture accounts for the production of more than 1 billion tons of manure annually (1). Manure can contain numerous animal-specific and zoonotic (transferable between animals and humans) pathogens, including bacteria such as Salmonella, pathogenic Escherichia coli, Campylobacter, Leptospira spp., Listeria monocytogenes, vancomycin-resistant enterococci (VRE), and methicillin-resistant Staphylococcus aureus (MRSA) and zoonotic protozoa such as Cryptosporidium and Giardia spp. (1-4). Additionally, although viruses tend to be host-specific pathogens, there is some evidence that swine influenza viruses, hepatitis E virus (HEV), and possibly other porcine viruses may be transmitted to humans (4, 5). Thus, human and animal health is a common concern if swine (or other animal) manures are not properly managed (1, 4).Untreated manure is applied to land as fertilizer in agricultural settings; most U.S. states require manure application practices that minimize contamination of surface water or groundwater (1). Nevertheless, manure can enter water through inadvertent mechanisms, including equipment failures, overapplication, runoff from open feedlots, storage overflow, accidents with manuretransporting equipment, se...

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Section: Resultsmentioning

confidence: 99%

Section: 6-25%mentioning

confidence: 99%

Section: 6-25%mentioning

confidence: 99%

Section: 6-25%mentioning

confidence: 99%

Section: 6-25%mentioning

confidence: 99%

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Genes Indicative of Zoonotic and Swine Pathogens Are Persistent in Stream Water and Sediment following a Swine Manure Spill

Haack

Duris

Kolpin

et al. 2015

Appl Environ Microbiol

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Microbial source tracking and land use associations for antibiotic resistance genes in private wells influenced by human and livestock fecal sources

et al. 2023

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Antimicrobial resistance is a growing public health problem that requires an integrated approach among human, agricultural, and environmental sectors. However, few studies address all three components simultaneously. We investigated the occurrence of five antibiotic resistance genes (ARGs) and the class 1 integron gene (intI1) in private wells drawing water from a vulnerable aquifer influenced by residential septic systems and land‐applied dairy manure. Samples (n = 138) were collected across four seasons from a randomized sample of private wells in Kewaunee County, Wisconsin. Measurements of ARGs and intI1 were related to microbial source tracking (MST) markers specific to human and bovine feces; they were also related to 54 risk factors for contamination representing land use, rainfall, hydrogeology, and well construction. ARGs and intI1 occurred in 5%–40% of samples depending on target. Detection frequencies for ARGs and intI1 were lowest in the absence of human and bovine MST markers (1%–30%), highest when co‐occurring with human and bovine markers together (11%–78%), and intermediate when co‐occurring with just one type of MST marker (4%–46%). Gene targets were associated with septic system density more often than agricultural land, potentially because of the variable presence of manure on the landscape. Determining ARG prevalence in a rural setting with mixed land use allowed an assessment of the relative contribution of human and bovine fecal sources. Because fecal sources co‐occurred with ARGs at similar rates, interventions intended to reduce ARG occurrence may be most effective if both sources are considered.

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Leaching of viruses and other microorganisms naturally occurring in pig slurry to tile drains on a well-structured loamy field in Denmark

et al. 2017

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The amount of animal manure used in modern agriculture is increasing due to the increase in global animal production. Pig slurry is known to contain zoonotic bacteria such as E. coli, Salmonella spp. and Campylobacter spp., and viruses such as hepatitis E virus and group A rotavirus. Coliform bacteria, present in manure, have previously been shown to leach into tile drains. This poses a potential threat to aquatic environments and may also influence the quality of drinking water. As knowledge is especially scarce about the fate of viruses when applied to fields in natural settings, this project sets out to investigate the leaching potential of six different microorganisms: E. coli and Enterococcus spp. (detected by colony assay), somatic coliphages (using plaque assays), and hepatitis E virus, porcine circovirus type 2, and group A rotavirus (by real-time polymerase chain reaction). All six microorganisms leached through the soil entering the tile drains situated at 1-m depth the first day following pig slurry application. The leaching pattern of group A rotavirus differed substantially from the pattern for somatic coliphages, which are otherwise used as indicators for virus contamination. Furthermore, group A rotavirus was detected in monitoring wells at 3.5-m depth up to 2 months after pig slurry application. The detection of viral genomic material in drainage water and shallow groundwater signifies a potential hazard to human health that needs to be investigated further, as water reservoirs used for recreational use and drinking water are potentially contaminated with zoonotic pathogens.

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Detection of Bacterial Indicators and Human and Bovine Enteric Viruses in Surface Water and Groundwater Sources Potentially Impacted by Animal and Human Wastes in Lower Yakima Valley, Washington

Cited by 42 publications

References 26 publications

Genes Indicative of Zoonotic and Swine Pathogens Are Persistent in Stream Water and Sediment following a Swine Manure Spill

Genes Indicative of Zoonotic and Swine Pathogens Are Persistent in Stream Water and Sediment following a Swine Manure Spill

Microbial source tracking and land use associations for antibiotic resistance genes in private wells influenced by human and livestock fecal sources

Leaching of viruses and other microorganisms naturally occurring in pig slurry to tile drains on a well-structured loamy field in Denmark

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