E-Book 2nd Congress

  • Detection of Leptospira spp., from sheep and goats blood in West Azerbaijan province, Iran
  • Ahmad Enferadi,1,* Abdolghaffar Ownagh,2 Amir Tukmechi,3 Mohsen soltani,4 Peyman Khademi,5 Amin Jaidari,6
    1. Urmia university
    2. Urmia University
    3. Urmia university
    4. Urmia university
    5. UrmiaUniversity
    6. Lorestan University


  • Introduction: Leptospirosis, an infectious disease that affects both humans and animals, is recognized as one of the most widespread zoonosis worldwide. Annually, an estimated one-half million cases of severe leptospirosis are reported globally. However, this number is probably underestimated because of the lack of reported cases and the misdiagnosis of this disease in many countries. Leptospirosis is caused by the pathogenic strains of the bacterium Leptospira. Currently, there are nearly 300 known serovars, and most of them have their primary reservoirs in wild and domestic animals, of which rodents and rats are the most common source worldwide. Infected rats shed Leptospira spp., in their urine over an extended period of time. Humans and animals get infected through direct or indirect contact with urine, water, or soil contaminated by Leptospira spp. Approximately one-half of the pathogenic serovars belong to L. interrogans or L. borgpetersenii. Classically, the diagnosis of leptospirosis is based on serological tests, such as the microscopic agglutination test (MAT). In this test, reaction takes place between a leptospiral isolate and reference hyperimmune rabbit antisera. However, this method is laborious and time-consuming, and it requires extensive collection of reference strains and their corresponding rabbit antisera. Various molecular approaches have been developed, such as polymerase chain reaction (PCR)-based methods, to improve the diagnosis of leptospirosis. PCR has been successfully applied as a rapid, sensitive, and specific tool for the detection of several microorganisms, including Leptospira, in a variety of specimens from different hosts. The rapidity and reproducibility of pulsed-field gel electrophoresis (PFGE) makes it a very useful technique for typing Leptospira strains.
  • Methods: A number of 215 blood samples were randomly collected from sheep (n = 117) and goats (n = 98) belonged to 15 sheep and goats' flocks randomly selected in three different geographical regions of west Azerbaijan (north, center and south) during four seasons in 2022. A number of ten flocks from each region, including five flocks of sheep and goats were randomly selected and 25 animals were sampled from each flock. Sampling of apparently healthy animals was performed. Each blood sample was collected from the jugular vein of the sheep and goat into a sterile tube containing an anticoagulant (ethylene diamine tetra acetic acid, EDTA).The collected blood samples were placed on ice and immediately transferred to the microbiology laboratory at the Faculty of Veterinary Medicine. DNA extraction from re-suspended pellet was performed using Blood Genomic DNA Extraction Mini Kit, (Favorgen-Taiwan) according to the kit's manufacturer instructions. The quality and quantity of the extracted DNA was evaluated by Nano Drop 2000c (Termo Scientific, USA). The extracted DNA from samples was kept at −20 °C for the later use in Nested PCR. During DNA extraction procedure, elution buffer from the extraction kit was used as Negative Control of Extraction. Primer for Nested PCR F- CATGCAAGTCAAGCGGAGTA FN- ACGCCCAATGATTCCGAACA R- AGTTGAGCCCGCAGTTTTC RN- TTCGGCCACAATGGAACTGAGA
  • Results: The percentage of positive blood samples for Leptospira spp., in sheep and goat was by using 16S rRNA gene 16.2% and 8.1% respectively. Prevalence of Leptospira spp., in the blood samples collected from sheep and goat dairy farms from West Azerbaijan Province. Animal Sheep (No.117) Goat (No.98) p-value Gene 16SrRNA 16SrRNA Genus female 13/77 (16.8%) 5/67 (7.4%) P < 0.05 male 6/40 (15%) 3/31 (9.6%) Total 19/117 (16.2%) 8/98 (8.1%) P < 0.05
  • Conclusion: It was concluded that sheep and goats can play an important role in the epidemiology of Leptospiral disease as the reservoir for Leptospira spp. The molecular detection of Leptospira spp., using Nested-PCR method in blood samples showed that PCR can be used an easy and reliable approach for detecting Leptospira spp. According to the reported results, the prevalence of Leptospira spp., was higher in sheep blood than goat blood.
  • Keywords: Leptospira spp., Nested-PCR, blood, sheep, goat