Abstract

In the past decade, two leptospirosis outbreaks occurred among strawberry harvesters in Germany, with 13, and 45 reported cases respectively. In both outbreaks, common voles (Microtus arvalis) infected with Leptospira kischneri serovar Grippotyphosa were identified as the most likely outbreak source. In an univariate analysis, eating unwashed strawberries was identified as one of the risk factors associated with Leptospira infection. The aim of this study was to evaluate the survival time of L. kirschneri serovar Grippotyphosa on strawberries under varying conditions. Strawberries were spiked with 5x109 of both a laboratory reference strain (strain Moskva V) and an outbreak field strain (94-6/2007) of L. kirschneri serovar Grippotyphosa sequence type 110. Survival times were investigated in a fully crossed design with three incubation times (2h, 4h, 6h and 8h) and three temperatures (15°C, 21°C and 25°C) with three replicated for each condition. A wash protocol was developed and recovered Leptospira were determined by qPCR, dark field microscopy and culturing. Viable L. kirschneri of both the reference strain and the field strain were identified in all samples at 25°C and an incubation time of 2h, but only 1/9 (11%) and 4/9 (44%) of the samples incubated at 15°C were positive, respectively. Both reference and field strain were viable only in 2/9 (22%) at 25° after 6h. After an 8h incubation, viable Leptospira could not be identified on the surface of the strawberries or within the fruit for any of the tested conditions. Based on these results, the exposure risk of consumers to viable Leptospira spp. through the consumption of strawberries bought at the retail level is most likely very low. However, there is a potential risk of Leptospira infection by consumption of strawberries on pick-your-own farms.

Highlights

  • Leptospirosis is a zoonotic disease, which occurs worldwide in both humans and animals [1, 2]

  • Both a laboratory and a field strain (94-6/2007) of L. kirschneri serovar Grippotyphosa sequence type 110 were used in this study

  • Washing the strawberries in flasks showed the highest recovery of 3.1x106 genome equivalents (GE)/ml (94% compared to the reference sample) by quantification using qPCR and 4.6x107 cells/ml (99%) using microscopy (Fig 1)

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Summary

Introduction

Leptospirosis is a zoonotic disease, which occurs worldwide in both humans and animals [1, 2]. Pathogenic leptospires colonize the kidneys of animal hosts and are shed with the urine. Humans and animals are mostly infected through urine contaminated water or soil which come in contact with abrasions or cuts in the skin or mucous membranes [3].

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