Survival of the green fluorescent protein-transformed human pathogens Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium was studied in a laboratory-simulated lettuce production chain. Dairy cows were fed three different roughage types: high-digestible grass silage plus maize silage (6:4), low-digestible grass silage, and straw. Each was adjusted with supplemental concentrates to high and low crude protein levels. The pathogens were added to manure, which was subsequently mixed (after 56 and 28 days for E. coli O157:H7 and Salmonella serovar Typhimurium, respectively) with two pairs of organically and conventionally managed loamy and sandy soil. After another 14 days, iceberg lettuce seedlings were planted and then checked for pathogens after 21 days of growth. Survival data were fitted to a logistic decline function (exponential for E. coli O157:H7 in soil). Roughage type significantly influenced the rate of decline of E. coli O157:H7 in manure, with the fastest decline in manure from the pure straw diet and the slowest in manure from the diet of grass silage plus maize silage. Roughage type showed no effect on the rate of decline of Salmonella serovar Typhimurium, although decline was significantly faster in the manure derived from straw than in the manure from the diet of grass silage plus maize silage. The pH and fiber content of the manure were significant explanatory factors and were positively correlated with the rate of decline. With E. coli O157:H7 there was a trend of faster decline in organic than in conventional soils. No pathogens were detected in the edible lettuce parts. The results indicate that cattle diet and soil management are important factors with respect to the survival of human pathogens in the environment.

The recent increase in foodborne disease associated with the consumption of fresh vegetables stresses the importance of the development of intervention strategies that minimize the risk of preharvest contamination. To identify risk factors for Escherichia coli O157:H7 persistence in soil, we studied the survival of a Shiga-toxin-deficient mutant in a set of 36 Dutch arable manure-amended soils (organic/conventional, sand/loam) and measured an array of biotic and abiotic manure-amended soil characteristics. The Weibull model, which is the cumulative form of the underlying distribution of individual inactivation kinetics, proved to be a suitable model for describing the decline of E. coli O157:H7. The survival curves generally showed a concave curvature, indicating changes in biological stress over time. The calculated time to reach the detection limit ttd ranged from 54 to 105 days, and the variability followed a logistic distribution. Due to large variation among soils of each management type, no differences were observed between organic and conventional soils. Although the initial decline was faster in sandy soils, no significant differences were observed in ttd between both sandy and loamy soils. With sandy, loamy and conventional soils, the variation in ttd was best explained by the level of dissolved organic carbon per unit biomass carbon DOC/biomC, with prolonged survival at increasing DOC/biomC. With organic soils, the variation in ttd was best explained by the level of dissolved organic nitrogen (positive relation) and the microbial species diversity as determined by denaturing gradient gel electrophoresis (negative relation). Survival increased with a field history of low-quality manure (artificial fertilizer and slurry) compared with high-quality manure application (farmyard manure and compost). We conclude that E. coli O157:H7 populations decline faster under more oligotrophic soil conditions, which can be achieved by the use of organic fertilizer with a relatively high C/N ratio and consequently a relatively low rate of nutrient release.

No abstract

Manure samples were collected from 16 organic (ORG) and 9 low-input conventional (LIC) Dutch dairy farms during August and September 2004 to determine the prevalence of the STEC virulence genes stx 1 (encoding Shiga toxin 1), stx 2 (encoding Shiga toxin 2), and eaeA (encoding intimin), as well as the rfbE gene, which is specific for Escherichia coli O157. The rfbE gene was present at 52% of the farms. The prevalence of rfbE was higher at ORG farms (61%) than at LIC farms (36%), but this was not significant. Relatively more LIC farms were positive for all Shiga toxin-producing E. coli (STEC) virulence genes eaeA, stx 1 , and stx 2 , which form a potentially highly virulent combination. Species richness of Enterobacteriaceae, as determined by DGGE, was significantly lower in manure positive for rfbE. Survival of a green fluorescent protein-expressing E. coli O157:H7 strain was studied in the manure from all farms from which samples were obtained and was modeled by a biphasic decline model. The time needed to reach the detection limit was predominantly determined by the level of native coliforms and the pH (both negative relationships). Initial decline was faster for ORG manure but leveled off earlier, resulting in longer survival than in LIC manure. Although the nonlinear decline curve could theoretically be explained as the cumulative distribution of an underlying distribution of decline kinetics, it is proposed that the observed nonlinear biphasic pattern of the survival curve is the result of changing nutrient status of the manure over time (and thereby changing competition pressure), instead of the presence of subpopulations differing in the level of resistance.Shiga-toxin producing Escherichia coli (STEC) is an important group of zoonotic human pathogens, with E. coli O157 being the best known and most studied serotype (10). STEC strains are generally carried asymptomatically by cattle and shed in their feces, which in turn may serve as a means of maintenance and spread of these pathogens among cattle herds (47). Since the reported increase in food-borne disease associated with the consumption of fresh vegetables (51, 54), the potential contamination of vegetable crops that are grown in fields enriched with manure has raised increasing concern (18, 29, 52). Understanding on-farm survival and spread of human pathogens is of utmost importance in preventing the spread of this organism to the environment, groundwater, food, and crops and subsequently back to cattle.A considerable body of literature exists on the prevalence of STEC on cattle farms and statistically associated management factors which could be used for potential intervention strategies to reduce shedding of these pathogens (2, 53, 57). However, these studies are not unambiguous and have resulted in the identification of only a very few risk factors, like season and cattle age, which are ubiquitous. The direction of influence of the majority of the management practices like feeding regimen and housing conditions remains unclear and is under debate.D...

No abstract