Chapter 1
Isolation and Culture of Leptospira from Clinical
and Environmental Samples
Cyrille Goarant, Dominique Girault, Roman Thibeaux,
and Marie-Estelle Soupe ´ -Gilbert
Abstract
Leptospires, the etiological agents of leptospirosis, are fastidious slow-growing organisms. Here we
describe the isolation and routine maintenance of leptospires from clinical (blood, urine, or tissue) and
environmental (water or soil) samples. Using combinations of filtration, agar plating, and selective agents,
leptospires can be isolated in pure cultures even from complex contaminated sources in standard EMJH
culture medium.
Key words Leptospira, Isolation, Culture, Filtration, Selective agents, Soil, Water, Blood, Urine,
Kidney
1 Introduction
Culture and isolation of Leptospira has long been a reference technique
to biologically confirm acute leptospirosis in humans and animals as
well as to evidence chronic shedding by animal reservoirs [1]. Successful culture of virulent leptospires was achieved in 1915 using complex
culture media [2]. Only half a century later it was shown that leptospires could be cultured in a serum-free medium, leading to the development of the most widely used medium: the “EMJH” medium,
named from Ellinghausen, McCullough, Johnson and Harris
[3, 4]. Because leptospires are fastidious and slow-growing organisms,
the chemical inhibition of contaminants was also considered at the
same period in the 1960s [5, 6]. The advent of molecular techniques
[7–9] has revolutionized the diagnosis turnaround time, leading to
disregard Leptospira culture and isolation for diagnosis. In recent years
however, the development of whole genome sequencing and genomewide molecular studies has highlighted the usefulness of culture isolation [10], leading to a renewed interest in isolating leptospires from
clinical, but also environmental samples [11]. Here, we summarize the
Nobuo Koizumi and Mathieu Picardeau (eds.), Leptospira spp.: Methods and Protocols, Methods in Molecular Biology, vol. 2134,
https://doi.org/10.1007/978-1-0716-0459-5_1, © Springer Science+Business Media, LLC, part of Springer Nature 2020
1
Isolation and Culture of Leptospira from Clinical
and Environmental Samples
Cyrille Goarant, Dominique Girault, Roman Thibeaux,
and Marie-Estelle Soupe ´ -Gilbert
Abstract
Leptospires, the etiological agents of leptospirosis, are fastidious slow-growing organisms. Here we
describe the isolation and routine maintenance of leptospires from clinical (blood, urine, or tissue) and
environmental (water or soil) samples. Using combinations of filtration, agar plating, and selective agents,
leptospires can be isolated in pure cultures even from complex contaminated sources in standard EMJH
culture medium.
Key words Leptospira, Isolation, Culture, Filtration, Selective agents, Soil, Water, Blood, Urine,
Kidney
1 Introduction
Culture and isolation of Leptospira has long been a reference technique
to biologically confirm acute leptospirosis in humans and animals as
well as to evidence chronic shedding by animal reservoirs [1]. Successful culture of virulent leptospires was achieved in 1915 using complex
culture media [2]. Only half a century later it was shown that leptospires could be cultured in a serum-free medium, leading to the development of the most widely used medium: the “EMJH” medium,
named from Ellinghausen, McCullough, Johnson and Harris
[3, 4]. Because leptospires are fastidious and slow-growing organisms,
the chemical inhibition of contaminants was also considered at the
same period in the 1960s [5, 6]. The advent of molecular techniques
[7–9] has revolutionized the diagnosis turnaround time, leading to
disregard Leptospira culture and isolation for diagnosis. In recent years
however, the development of whole genome sequencing and genomewide molecular studies has highlighted the usefulness of culture isolation [10], leading to a renewed interest in isolating leptospires from
clinical, but also environmental samples [11]. Here, we summarize the
Nobuo Koizumi and Mathieu Picardeau (eds.), Leptospira spp.: Methods and Protocols, Methods in Molecular Biology, vol. 2134,
https://doi.org/10.1007/978-1-0716-0459-5_1, © Springer Science+Business Media, LLC, part of Springer Nature 2020
1