Browsing by Author "Weller, Daniel L."
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- Factors Associated With E. coli Levels in and Salmonella Contamination of Agricultural Water Differed Between North and South Florida WaterwaysMurphy, Claire M.; Strawn, Laura K.; Chapin, Travis K.; McEgan, Rachel; Gopidi, Sweeya; Friedrich, Loretta; Goodridge, Lawrence D.; Weller, Daniel L.; Schneider, Keith R.; Danyluk, Michelle D. (Frontiers, 2022-02-02)The microbial quality of agricultural water is often assessed using fecal indicator bacteria (FIB) and physicochemical parameters. The presence, direction, and strength of associations between microbial and physicochemical parameters, and the presence of human pathogens in surface water vary across space (e.g., region) and time. This study was undertaken to understand these associations in two produce-growing regions in Florida, USA, and to examine the pathogen ecology in waterways used for produce production. The relationship between Salmonella presence, and microbial and physicochemical water quality; as well as weather and land use factors were evaluated. Water samples were collected from six sites in North Florida (N = 72 samples) and eight sites in South Florida (N = 96 samples) over 12 sampling months. Land use around each sampling site was characterized, and weather and water quality data were collected at each sampling. Salmonella, generic Escherichia coli, total coliform, and aerobic plate count bacteria populations were enumerated in each sample. Univariable and multivariable regression models were then developed to characterize associations between microbial water quality (i.e., E. coli levels and Salmonella presence), and water quality, weather, and land use factors separately for North and South Florida. The E. coli and total coliforms mean concentrations (log(10) MPN/100 mL) were 1.8 +/- 0.6 and >3.0 +/- 0.4 in North and 1.3 +/- 0.6 and >3.3 +/- 0.2 in South Florida waterways, respectively. While Salmonella was detected in 23.6% (17/72) of North Florida and 28.1% (27/96) of South Florida samples, the concentration ranged between <0.48 and 1.4 log(10) MPN/100 mL in North Florida, and E. coli levels, and if a sample was Salmonella-positive. The factors associated with Salmonella presence and log(10) E. coli levels in North Florida differed from those in South Florida; no factors retrained in multivariable regression models were the same for the North and South Florida models. The differences in associations between regions highlight the complexity of understanding pathogen ecology in freshwater environments and suggest substantial differences between intra-state regions in risk factors for Salmonella contamination of agricultural water.
- Humans and Hoofed Livestock Are the Main Sources of Fecal Contamination of Rivers Used for Crop Irrigation: A Microbial Source Tracking ApproachDiaz-Gavidia, Constanza; Barria, Carla; Weller, Daniel L.; Salgado-Caxito, Marilia; Estrada, Erika M.; Araya, Anibal; Vera, Leonardo; Smith, Woutrina; Kim, Minji; Moreno-Switt, Andrea I.; Olivares-Pacheco, Jorge; Adell, Aiko D. (Frontiers, 2022-06-30)Freshwater bodies receive waste, feces, and fecal microorganisms from agricultural, urban, and natural activities. In this study, the probable sources of fecal contamination were determined. Also, antibiotic resistant bacteria (ARB) were detected in the two main rivers of central Chile. Surface water samples were collected from 12 sampling sites in the Maipo (n = 8) and Maule Rivers (n = 4) every 3 months, from August 2017 until April 2019. To determine the fecal contamination level, fecal coliforms were quantified using the most probable number (MPN) method and the source of fecal contamination was determined by Microbial Source Tracking (MST) using the Cryptosporidium and Giardia genotyping method. Separately, to determine if antimicrobial resistance bacteria (AMB) were present in the rivers, Escherichia coli and environmental bacteria were isolated, and the antibiotic susceptibility profile was determined. Fecal coliform levels in the Maule and Maipo Rivers ranged between 1 and 130 MPN/100-ml, and 2 and 30,000 MPN/100-ml, respectively. Based on the MST results using Cryptosporidium and Giardia host-specific species, human, cattle, birds, and/or dogs hosts were the probable sources of fecal contamination in both rivers, with human and cattle host-specific species being more frequently detected. Conditional tree analysis indicated that coliform levels were significantly associated with the river system (Maipo versus Maule), land use, and season. Fecal coliform levels were significantly (p < 0.006) higher at urban and agricultural sites than at sites immediately downstream of treatment centers, livestock areas, or natural areas. Three out of eight (37.5%) E. coli isolates presented a multidrug-resistance (MDR) phenotype. Similarly, 6.6% (117/1768) and 5.1% (44/863) of environmental isolates, in Maipo and Maule River showed and MDR phenotype. Efforts to reduce fecal discharge into these rivers should thus focus on agriculture and urban land uses as these areas were contributing the most and more frequently to fecal contamination into the rivers, while human and cattle fecal discharges were identified as the most likely source of this fecal contamination by the MST approach. This information can be used to design better mitigation strategies, thereby reducing the burden of waterborne diseases and AMR in Central Chile.
- Land Use, Weather, and Water Quality Factors Associated With Fecal Contamination of Northeastern Streams That Span an Urban-Rural GradientWeller, Daniel L.; Murphy, Claire M.; Johnson, Stephanie; Green, Hyatt; Michalenko, Edward M.; Love, Tanzy, M.T.; Strawn, Laura K. (Frontiers, 2022-02)Fecal contamination of surface water has been associated with multiple enteric disease outbreaks and food recalls. Thus, it is important to understand factors associated with fecal contamination of agricultural water sources. Since fecal indicator bacteria (FIB) were used to monitor surface water for potential fecal contamination, the purpose of the present study was to characterize associations between environmental factors, and (i) FIB (E. coli, Enterococcus, and coliform) levels, and (ii) host-specific fecal marker detection. This study used data collected from 224 sites along 3 waterways, which spanned an urban-rural gradient around Syracuse, New York. Between 2008 and 2017, 2,816 water samples were collected, and E. coli, Enterococcus, and/or coliform concentrations were enumerated. Thirty-one samples were also tested for human and ruminant microbial source-tracking markers. Water quality (e.g., turbidity, nitrate) and weather data were also collected for each site. Univariable Bayesian regression was used to characterize the relationship between each microbial target and land use, water quality, and weather factor. For eachmodel, probability of direction and region of practical equivalence overlap (ROPE) were calculated to characterize the association’s direction and strength, respectively. While levels of different FIB were not correlated with each other, FIB levels were associated with environmental conditions. Specifically, FIB levels were also positively associated with temperature, nutrient and sediment levels. Log10 E. coli levels increased by 0.20 (CI = 0.11, 0.31) and log10 Enterococcus levels increased by 0.68 (CI=0.08, 1.24) for each log10 increase in salinity and nitrate, respectively. These findings may indicate that similar processes drove microbial, sediment, and nutrient contamination of the sampled watersheds. While fecal contamination was strongly associated with land use, the direction of association varied between FIBs and the buffer distance used to calculate land use metrics. E. coli levels and human marker detection were positively associated with percent pasture cover within 122, 366, and 1,098mof the sampling site, while Enterococcus and coliform levels were only associated with pasture cover within 1,098m (not 122 or 366m). Ruminant markers were positively associated with pasture cover within 122m, but not 366 or 1,098m. These findings highlight the importance of considering (i) adjacent land use (and associated non-point sources of contamination) when developing strategies for managing fecal hazards associated in agricultural and recreational water, and (ii) spatial scale (e.g., 122 vs. 1,098m) when developing these strategies.
- Salmonella Prevalence Is Strongly Associated with Spatial Factors while Listeria monocytogenes Prevalence Is Strongly Associated with Temporal Factors on Virginia Produce FarmsMurphy, Claire M.; Weller, Daniel L.; Strawn, Laura K. (American Society for Microbiology, 2023-02-02)The heterogeneity of produce production environments complicates the development of universal strategies for managing preharvest produce safety risks. Understanding pathogen ecology in different produce-growing regions is important for developing targeted mitigation strategies. This study aimed to identify environmental and spatiotemporal factors associated with isolating Salmonella and Listeria from environmental samples collected from 10 Virginia produce farms. Soil (n = 400), drag swab (n = 400), and irrigation water (n = 120) samples were tested for Salmonella and Listeria, and results were confirmed by PCR. Salmonella serovar and Listeria species were identified by the Kauffmann-White-Le Minor scheme and partial sigB sequencing, respectively. Conditional forest analysis and Bayesian mixed models were used to characterize associations between environmental factors and the likelihood of isolating Salmonella, Listeria monocytogenes (LM), and other targets (e.g., Listeria spp. and Salmonella enterica serovar Newport). Surrogate trees were used to visualize hierarchical associations identified by the forest analyses. Salmonella and LM prevalence was 5.3% (49/920) and 2.3% (21/920), respectively. The likelihood of isolating Salmonella was highest in water samples collected from the Eastern Shore of Virginia with a dew point of >9.4°C. The likelihood of isolating LM was highest in water samples collected in winter from sites where <36% of the land use within 122 m was forest wetland cover. Conditional forest results were consistent with the mixed models, which also found that the likelihood of detecting Salmonella and LM differed between sample type, region, and season. These findings identified factors that increased the likelihood of isolating Salmonella- and LM-positive samples in produce production environments and support preharvest mitigation strategies on a regional scale. IMPORTANCE This study sought to examine different growing regions across the state of Virginia and to determine how factors associated with pathogen prevalence may differ between regions. Spatial and temporal data were modeled to identify factors associated with an increased pathogen likelihood in various on-farm sources. The findings of the study show that prevalence of Salmonella and L. monocytogenes is low overall in the produce preharvest environment but does vary by space (e.g., region in Virginia) and time (e.g., season), and the likelihood of pathogen-positive samples is influenced by different spatial and temporal factors. Therefore, the results support regional or scale-dependent food safety standards and guidance documents for controlling hazards to minimize risk. This study also suggests that water source assessments are important tools for developing monitoring programs and mitigation measures, as spatiotemporal factors differ on a regional scale.
- Spatial Versus Nonspatial Variance in Fecal Indicator Bacteria Differs Within and Between PondsMurphy, Claire M.; Weller, Daniel L.; Ovissipour, Reza; Boyer, Renee R.; Strawn, Laura K. (Elsevier, 2023-03)Surface water environments are inherently heterogenous, and little is known about variation in microbial water quality between locations. This study sought to understand how microbial water quality differs within and between Virginia ponds. Grab samples were collected twice per week from 30 sampling sites across nine Virginia ponds (n = 600). Samples (100 mL) were enumerated for total coliform (TC) and Escherichia coli (EC) levels, and physicochemical, weather, and environmental data were collected. Bayesian models of coregionalization were used to quantify the variance in TC and EC levels attributable to spatial (e.g., site, pond) versus nonspatial (e.g., date, pH) sources. Mixed-effects Bayesian regressions and conditional inference trees were used to characterize relationships between data and TC or EC levels. Analyses were performed separately for each pond with ≥3 sampling sites (5 intrapond) while one interpond model was developed using data from all sampling sites and all ponds. More variance in TC levels were attributable to spatial opposed to nonspatial sources for the interpond model (variance ratio [VR] = 1.55) while intrapond models were pond dependent (VR: 0.65–18.89). For EC levels, more variance was attributable to spatial sources in the interpond model (VR = 1.62), compared to all intrapond models (VR < 1.0) suggesting that more variance is attributable to nonspatial factors within individual ponds and spatial factors when multiple ponds are considered. Within each pond, TC and EC levels were spatially independent for sites 56–87 m apart, indicating that different sites within the same pond represent different water quality for risk management. Rainfall was positively and pH negatively associated with TC and EC levels in both inter- and intrapond models. For all other factors, the direction and strength of associations varied. Factors driving microbial dynamics in ponds appear to be pond-specific and differ depending on the spatial scale considered.
- Strain, Soil-Type, Irrigation Regimen, and Poultry Litter Influence Salmonella Survival and Die-off in Agricultural SoilsBardsley, Cameron; Weller, Daniel L.; Ingram, David T.; Chen, Yuhuan; Oryang, David O.; Rideout, Steven L.; Strawn, Laura K. (2021-03-16)The use of untreated biological soil amendments of animal origin (BSAAO) have been identified as one potential mechanism for the dissemination and persistence of Salmonella in the produce growing environment. Data on factors influencing Salmonella concentration in amended soils are therefore needed. The objectives here were to (i) compare die-off between 12 Salmonella strains following inoculation in amended soil and (ii) characterize any significant effects associated with soil-type, irrigation regimen, and amendment on Salmonella survival and die-off. Three greenhouse trials were performed using a randomized complete block design. Each strain (similar to 4 log CFU/g) was homogenized with amended or non-amended sandy-loam or clay-loam soil. Salmonella levels were enumerated In 25 g samples 0, 0.167 (4 h), 1,2, 4, 7, 10, 14, 21,28, 56, 84, 112, 168, 210, 252, and 336 days post-inoculation (dpi), or until two consecutive samples were enrichment negative. Regression analysis was performed between strain, soil-type, Irrigation, and (i) time to last detect (survival) and (li) concentration at each time-point (die-off rate). Similar effects of strain, irrigation, soil-type, and amendment were identified using the survival and die-off models. Strain explained up to 18% of the variance in survival, and up to 19% of variance In die-off rate. On average Salmonella survived for 129 days in amended soils, however, Salmonella survived, on average, 30 days longer In clay-loam soils than sandy-loam soils [95% Confidence interval (Cl) = 45, 15], with survival time ranging from 84 to 210 days for the individual strains during dally irrigation. When strain- specific associations were investigated using regression trees, S. Javiana and S. Saintpaul were found to survive longer In sandy-loam soil, whereas most of the other strains survived longer In clay-loam soil. Salmonella also survived, on average, 128 days longer when irrigated weekly, compared to daily (Cl = 101, 154), and 89 days longer in amended soils, than non-amended soils (Cl = 61,116). Overall, this study provides insight into Salmonella survival following contamination of field soils by BSAAO. Specifically, Salmonella survival may be strain- specific as affected by both soil characteristics and management practices. These data can assist in risk assessment and strain selection for use in challenge and validation studies.