With the increased consumption of fresh produce, a proportional increase in numbers of produce-related foodborne illness has been observed. An estimate of foodborne illness during 1998 to 2008 attributed ∼46% of the incidences to produce. Any foodborne illness associated with produce can have devastating consequences to the industry. The most recent data from the Centers for Disease Control and Prevention implicate leafy vegetables, vine-stalk vegetables, root vegetables, and sprouts as the most common cause of produce-related foodborne outbreaks. Excess rainfall or flooding, mainly by altering levels of soil moisture and oxygen content, affects the microbial community in soil. The goal of this research was to determine the survivability of a three-serovar Escherichia coli and a five-serovar Salmonella enterica cocktail in microcosms prepared with Candler sand (CS) and Orangeburg sandy loam (OSL) soils. Microcosms were prepared with low, medium, and high volumetric water contents and were incubated at 20 and 30°C. Serotyping was used to determine which E. coli or Salmonella serovar(s) from each cocktail persisted. Microcosm inoculation levels were ∼7.0 log CFU/g. Sampling for CS and OSL microcosms incubated at 20°C ended on day 364 and 357, respectively. The reduction of Salmonella and E. coli to below the limit of detection (extinction) in CS microcosms (incubated at 30°C at all volumetric water content [VWC] levels) was reached on day 168 and 56, respectively. Extinction of Salmonella and E. coli in OSL microcosms (incubated at 30°C at all VWCs) was reached on day 168 and 224, respectively. Of the Salmonella and E. coli serovars analyzed, Salmonella Javiana persisted the longest in both soil types, whereas E. coli O104:H4 and E. coli O145 persisted the longest in CS and OSL microcosms, respectively. Results from the current study suggest that soil type and temperature influenced pathogen persistence in CS and OSL soils more than moisture level and pathogen type.
For over a decade, Salmonella contamination has increasingly led to outbreaks of foodborne illness associated with fresh produce. The use of untreated animal manures, or biological soil amendments of animal origin (BSAAO), to amend agricultural soils holds a risk of contamination from foodborne pathogens, such as Salmonella . This study was conducted to determine the prevalence, concentration, serotypes, and antimicrobial resistance (AMR) profiles of Salmonella in poultry litter from Florida farms. Litter pH, total Kjeldahl nitrogen (TKN), total ammonia nitrogen (TAN), total phosphorus (P 2 O 5 ), total potassium (K 2 O), moisture content, total solids, total ash, organic matter, and aerobic plate count (APC) were also measured. Litter samples (n = 54) were collected from 18 broiler farms across three seasons (spring, summer, and winter). Salmonella concentrations were enumerated using a most probable number (MPN) method and antimicrobial susceptibility testing (AST) was performed. The prevalence of Salmonella in litter samples was 61.1%, with a geometric mean of 0.21 ± 20.7 MPN/g. Across all seasons, Salmonella concentrations were not influenced by the chemical, physical, or microbial properties measured. Recovered Salmonella isolates (n = 290) were grouped into serogroups O:4 (43.1%), O:7 (26.9%), O:8 (11.0%), O:1,3,10,19 (7.9%), and O:9,46 (7.2%). Serotyping select Salmonella isolates (n = 47) resulted in 12 serotypes, with the most common being Typhimurium (27.7%), Kentucky (17.0%), Enteritidis (14.9%), and Mbandaka (14.9%). Antimicrobial resistance to tetracycline (29.8%), sulfisoxazole (23.4%), and streptomycin (14.9%) was observed. No isolates were resistant to more than two antimicrobial agents. This study provides valuable information towards future risk assessments for the use of poultry litter as an untreated BSAAO.
We investigated the effects of chromate (CrVI) and sulfate on their uptake and translocation in As-hyperaccumulator Pteris vittata. Plants were exposed to 1) 0.1 mM CrVI and 0, 0.25, 1.25 or 2.5 mM sulfate or 2) 0.25 mM sulfate and 0, 0.5, 2.5 or 5.0 mM CrVI for 1 d in hydroponics. P. vittata accumulated 26 and 1261 mg kg(-1) Cr in the fronds and roots at CrVI0.1, and 2197 and 1589 mg kg(-1) S in the fronds and roots at S0.25. Increasing sulfate concentrations increased Cr root concentrations by 16-66% and helped CrVI reduction to CrIII whereas increasing CrVI concentrations increased frond sulfate concentrations by 3-27%. Increasing sulfate concentrations enhanced TBARS concentrations in the biomass, indicating oxidative stress caused lipid peroxidation in plant cell membranes. However, addition of 0.25-2.5 mM sulfate alleviated CrVI's toxic effects and decreased TBARS from 23.5 to 9.46-12.3 μmol g(-1) FW. Though CrVI was supplied, 78-96% of CrIII was in the biomass, indicating efficient CrVI reduction to CrIII by P. vittata. The data indicated the amazing ability of P. vittata in Cr uptake at 289 mg kg(-1) h(-1) with little translocation to the fronds. These results indicated that P. vittata had potential in Cr phytoremediation in contaminated sites but further studies are needed to evaluate this potential. The facts that CrVI and sulfate helped each other in uptake by P. vittata suggest that CrVI was not competing with sulfate uptake in P. vittata. However, the mechanisms of how sulfate and CrVI enhance each other's accumulation in P. vittata need further investigation.
Fresh produce outbreaks due to Shiga toxin-producing Escherichia coli (STEC) continue to occur in the United States (US). Manure-amended soils can pose a public health risk when used for growing raw agricultural commodities. Knowing the prevalence and concentration of STEC in untreated biological soil amendments of animal origin (BSAAO) is important to help guide the most appropriate pre-harvest interval(s) following application to limit risks from these soil amendments. Bovine manure samples were collected from 12 farms in Florida, including samples from piles, lagoons, barns, and screened solids. Two methods were used to detect stx1 / 2 and rfbE genes in samples. A prevalence rate of 9% for stx1 and/or stx2 and 19% for rfbE was observed from the 518 bovine manure samples evaluated. A most probable number (MPN) assay was performed on stx + samples when applicable. The geometric mean for stx+ samples (n = 20) was 3.37 MPN g -1 (0.53 log MPN g -1 ) with a maximum value of 6,800 MPN g -1 (3.83 log MPN g -1 ). This research was part of a larger nationwide geographical study on the prevalence and concentration of STEC in bovine manure to help guide regulations on feasible pre-harvest intervals for the application of untreated BSAAO.
While the soil microbiome may influence pathogen survival, determining the major contributors that reduce pathogen survival is inconclusive. This research was performed to determine the survival of E. coli O157 in autoclaved and natural (unautoclaved) sandy soils. Soils were inoculated with three different E. coli O157 strains (stx1+/stx2+, stx1-/stx2-, and stx1-/ stx2+), and enumerated until extinction at 30˚C. There was a significant difference in the survival of E. coli O157 based on soil treatment (autoclaved versus natural) at 30˚C on days 1 (P = 0.00022), 3, (P = 2.53e-14), 7 (P = 5.59e-16), 14 (P = 1.072e-12), 30 (P = 7.18e-9), and 56 (P = 0.00029), with greater survival in autoclaved soils. The time to extinction (two consecutive negative enrichments) for all three strains was 169 and 84 days for autoclaved and natural soils, respectively. A separate E. coli O157 trial supplemented with 16S rRNA gene sequencing of the soil microbiome was performed at 15˚C and 30˚C on days 0, 7, 14, and 28 for each soil treatment. Greater species richness (Chao1, P = 2.2e-16) and diversity (Shannon, P = 2.2e-16) was observed in natural soils in comparison with autoclaved soils. Weighted UniFrac (beta-diversity) showed a clear distinction between soil treatments (P = 0.001). The greatest reduction of E. coli O157 was observed in natural soils at 30˚C, and several bacterial taxa positively correlated (relative abundance) with time (day 0 to 28) in these soils (P < 0.05), suggesting that the presence of those bacteria might cause the reduction of E. coli O157. Taken together, a clear distinction in E. coli O157 survival, was observed between autoclaved and natural soils along with corresponding differences in microbial diversity in soil treatments. This research provides further insights into the bacterial taxa that may influence E. coli O157 in soils.
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