Evaluation of leech salivation on infectious opened-wound healing with Proteus mirabilis in rat

Document Type : Original Article

Authors
Seyedeh Shaghayegh Zenoozzadeh 1
Sahar Ghaffarikhaligh 2
Hamid Estaji 3
Hamidreza Moslemi 4
1 Graduated from Veterinary medicine School, Semnan University, Semnan, Iran.
2 Department of Pathobiology, Veterinary medicine School , Semnan University, Semnan, Iran.
3 Department of pathobiology, Veterinary medicine School , Semnan University, Semnan, Iran.
4 Department of clinical sciences, veterinary medicine school, Semnan University, Semnan, Iran.
10.61882/eltiamj.12.1.12
Abstract
Background: Infection control and improvement of the wound healing process are of great importance. Leech saliva has anti-inflammatory, antimicrobial, and restorative properties, so it can be a good alternative to common antibiotics resistant to many bacteria.
 Objectives: This study aimed to investigate the effect of leech saliva on the healing process of an open skin wound infected by Proteus mirabilis by examining histopathological and microbiological changes.
Methods: After preparing leech saliva, a 5 percent leech ointment (95 grams of Eucerin, 5 grams of leech saliva) was made, and 60 male Wistar rats were prepared and divided into 4 groups. A wound with a 1.5cm diameter was created by a biopsy punch in the posterior area between the two scapulae skin of each rat. Then, it was infected by inoculation of 106cfu of bacteria. Then, the groups were treated separately with gentamicin ointment, Eucerin, and leech ointment. One group didn’t receive any medication as a control. Ointments were renewed every other day for up to 21 days. For microbial and histopathological investigations, sterile samples were taken randomly from 5 rats on days 7, 14, and 21 after euthanizing.
 Results: In the microbial investigation of the total number of bacteria during 21days, the bacterial population in the group treated with leech saliva was less than the other groups, which had a significant difference with the control group, and specifically, the difference in the number of bacteria on the seventh day was significant with the other groups. In the examination of pathology sections, including the examination of collagen, epithelization, fibroblast density, and the amount of inflammation and angiogenesis, there was a significant difference between the treatment group with leech saliva and the control group in inflammation, epithelization, and collagen production, and no significant difference was observed between the leech and gentamicin groups in most of the parameters, except for the epithelizing factor, which had a significant difference on the 21st day of treatment.
Conclusions: This research aims to investigate leech saliva's effect in controlling inflammation and accelerating healing in infected wounds with Proteus mirabilis bacteria. The findings of this study show that leech saliva causes collagen production and increases the speed of the epithelialization process. Also, due to its antimicrobial properties, leech saliva can control the Proteus mirabilis bacterial population and can be a suitable substitute for antibiotics in short-term use. According to the discussion of antibiotic resistance regarding this bacterium, leech saliva can be used with appropriate antibiotics to control and accelerate the process of infected wounds, especially wounds infected with Proteus mirabilis.

Keywords

Subjects

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Eltiam
Volume 12, Issue 1 - Serial Number 23
November 2025
Pages 172-185

  • Receive Date 02 January 2025
  • Revise Date 01 February 2025
  • Accept Date 04 March 2026
  • Publish Date 22 November 2025