Past studies have shown that the on-farm distribution of Listeria monocytogenes is affected by environmental factors (e.g., weather). However, most studies were conducted at large scales (e.g., across farms), whereas few studies examined drivers of L. monocytogenes prevalence at smaller scales (e.g., within a single field). This study was performed to address this knowledge gap by (i) tracking L. monocytogenes distribution in two fields on one farm over a growing season and (ii) identifying factors associated with L. monocytogenes isolation from drag swab, soil, and agricultural water samples. Overall, L. monocytogenes was detected in 78% (21 of 27), 19% (7 of 36), and 8% (37 of 486) of water, drag swab, and soil samples, respectively. All isolates were characterized by pulsed-field gel electrophoresis. Of the 43 types identified, 14 were isolated on multiple sampling visits and/or from multiple sample types, indicating persistence in or repeated introduction into the farm environment during the study. Our findings also suggest that L. monocytogenes prevalence, even at the small spatial scale studied here, (i) was not uniform and (ii) varied more within fields than between fields or over time. This is illustrated by plot (in-field variation), field (between-field variation), and sampling visit (time), accounting for 18, 2, and 3% of variance in odds of isolating L. monocytogenes, respectively. Moreover, according to random forest analysis, water-related factors were among the top-ranked factors associated with L. monocytogenes isolation from all sample types. For example, the likelihood of isolating L. monocytogenes from drag and soil samples increased monotonically as rainfall increased. Overall, findings from this single-farm study suggests that mitigation strategies for L. monocytogenes in produce fields should focus on water-associated risk factors (e.g., rain and distance to water) and be tailored to specific high-risk in-field areas.

  • L. monocytogenes (LM) prevalence varied more over space than time.

  • Water samples had a higher LM prevalence than soil samples.

  • Despite high LM prevalence in environmental samples, produce contamination was rare.

  • Rain and distance to water were positively associated with LM isolation.

  • Repeated isolation of PFGE types suggests persistence or repeated introduction.

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