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Submitted: 15 Jun 2025
Revision: 28 Jul 2025
Accepted: 02 Aug 2025
ePublished: 22 Nov 2025
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Avicenna J Clin Microbiol Infect. 2025;12(4): 188-194.
doi: 10.34172/ajcmi.3712
  Abstract View: 11
  PDF Download: 6

Original Article

Isolation, Molecular Detection, and Sequence Analysis of Pathogenic Leptospira spp. in Paddy Field Water Samples in Amol, Northern Iran

Fereydoun Imani 1 ORCID logo, Esmail Fattahi 2* ORCID logo, Rahem Khoshbakht 3,4* ORCID logo, Sadegh Fattahi 5 ORCID logo, Mohsen Asouri 6 ORCID logo

1 Department of Microbiology, Am.C., Islamic Azad University, Amol, Iran
2 Department of Biology, Am.C., Islamic Azad University, Amol, Iran
3 Department of Pathobiology, Amol University of Special Modern Technologies, Amol, Iran
4 Zoonotic Diseases Research Group, Faculty of Veterinary Medicine, Amol University of Modern Technologies, Amol, Iran
5 North Research Center of Pasteur Institute, Amol, Iran
6 Department of Paramedicine, Amol School of Paramedicine, Mazandaran University of Medical Sciences, Sari, Iran
*Corresponding Authors: Esmail Fattahi, Email: esmail_fattahy@yahoo.com, Email: e.fattahi@iauamol.ac.ir; Rahem Khoshbakht, Email: r.khoshbakht@ausmt.ac.ir, Email: khoshbakht.r@gmail.com

Abstract

Background: Leptospirosis constitutes a zoonotic affliction prevalent in tropical and subtropical locales, instigated by the pathogenic strains of Leptospira. The objective of the present study was to isolate and detect Leptospira from water samples sourced from paddy fields located in Amol, northern Iran.

Methods: A total of 108 samples were procured from rice fields during the spring and summer of 2023. Cultivation, microscopy, and molecular analyses were employed to accurately identify L. interrogans. Both conventional and real-time polymerase chain reactions (RT-PCR) targeting the lipL32 gene were executed for the detection of L. interrogans. In addition, the sequence analysis of the sucA and gluM genes was performed after PCR.

Results: The findings revealed that 6/108 (5.55%) of the samples were culture-positive, with all cases corroborated through both RT-PCR and traditional PCR methods. A simple PCR assay showed positive results for 17/108 (15.74%) water samples. RT-PCR successfully identified 17/108 (15.74%) samples as positive for L. interrogans. The sequence analysis of the sucA gene from the water sample demonstrated high similarity to Leptospira weilii. Eventually, the sequence analysis of the gluM gene from two water samples displayed high similarity to Leptospira borgpetersenii.

Conclusion: This investigation highlights the efficacy of synergizing molecular techniques with traditional culture methodologies for the surveillance of Leptospira in areas characterized by an elevated risk. The current research serves as the inaugural report, delineating the emergence of leptospires from rice field samples, alongside the characterization of isolates obtained from culture.



Please cite this article as follows: Imani F, Fattahi E, Khoshbakht R, Fattahi S, Asouri M. Isolation, molecular detection, and sequence analysis of pathogenic leptospira spp. In paddy field water samples in Amol, northern Iran. Avicenna J Clin Microbiol Infect. 2025;12(4):188-194. doi:10.34172/ajcmi.3712
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