Abstract: Wounds on the skin are a common health issue affecting working equines. This study aimed to evaluate the in-vitro antibacterial properties of crude methanolic extracts from selected medicinal plants against pathogens isolated from equine skin wounds in Merti district. Agar well and disc diffusion tests were used to determine the mean zone of inhibition, while broth dilution methods were used to determine the minimum inhibitory concentrations and minimum bactericidal concentrations (MBCs), further confirming the potent antibacterial effects of the selected medicinal plant extracts. One way analysis of variance was used to compute the mean zone of inhibition (mm ± SEM) using SPSS version 20. The results showed that Staphylococcus aureus was the most frequently isolated bacterial pathogen. The medicinal plants Zingiber officinale (ginger), Allium sativum (garlic), Croton macrostachyus, and Solanum incanum exhibited significant antibacterial activity against S. aureus, Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae. On agar well diffusion method, the highest and lowest zone of inhibition was recorded in C. macrostachyus (28.0 ± 1.2 mm for S. aureus standard strain) and Z. officinale (12.7 ± 0.7 mm for K. pneumoniae wound isolate), respectively at 780 mg/mL concentration. Similarly, C. macrostachyus showed the highest (28.3 ± 0.9) zone of inhibition using the disc diffusion test for S. aureus standard strain. In combined extracts, the highest zone of inhibition was found against standard strain of S. aureus with Croton macrostachyus + Zingiber officinale (33.0 ± 0.6), followed by both Allium sativum + Zingiber officinale and Allium sativum + Croton macrostachyus at 32 ± 1.2 mm at 780 mg/mL concentration. The minimum inhibitory concentration ranged from 12.18 mg/mL for A. sativum (against standard S. aureus) to 390 mg/mL for S. incanum (against P. aeruginosa), while MBC ranged from 24.38 mg/mL for A. sativum (against S. aureus and E. coli) to > 390 mg/mL for S. incanum against K. pneumoniae and P. aeruginosa. The findings demonstrate the potential of these natural resources as alternative or complementary treatments for managing bacterial infections associated with equine skin wounds, which is particularly relevant in the context of the growing global challenge of antibiotic resistance, and provide valuable baseline information for future in-vivo investigations and the potential development of herbal-based wound management strategies in equine veterinary practice, warranting further in-vivo studies.
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Research Overview
This study investigated the types of bacteria found in equine skin wounds and tested how effective certain medicinal plant extracts are at killing these bacteria in laboratory conditions.
Background and Objectives
Equine skin wounds are common health issues that can be complicated by bacterial infections.
The study was conducted in the Merti district, focusing on working equines which are frequently affected by wounds.
The main aim was to isolate major bacterial pathogens from these wounds and evaluate the antibacterial activity of selected medicinal plants against these bacteria.
Methods Used
Bacterial isolation and identification from equine skin wounds.
Preparation of crude methanolic extracts from selected medicinal plants:
Zingiber officinale (ginger)
Allium sativum (garlic)
Croton macrostachyus
Solanum incanum
Antibacterial activity assessed using:
Agar well diffusion method
Disc diffusion test
Broth dilution method to determine Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)
Statistical analysis using one-way ANOVA with SPSS software to compare the zones of inhibition (measures of antibacterial effectiveness).
Key Findings on Bacterial Isolates
The most frequently isolated bacterial pathogen was Staphylococcus aureus.
Other isolated pathogens included:
Pseudomonas aeruginosa
Escherichia coli
Klebsiella pneumoniae
Antibacterial Activity of Medicinal Plants
All four medicinal plant extracts showed significant antibacterial effects against the isolated bacteria.
Specific observations from agar well diffusion:
Croton macrostachyus had the largest zone of inhibition (28.0 ± 1.2 mm) against the standard strain of S. aureus at 780 mg/mL concentration.
Zingiber officinale showed the smallest zone (12.7 ± 0.7 mm) against K. pneumoniae wound isolates at the same concentration.
Disc diffusion results confirmed a strong antibacterial effect of C. macrostachyus, particularly against standard S. aureus (28.3 ± 0.9 mm zone of inhibition).
Combined extracts exhibited even greater antibacterial activity:
Combination of C. macrostachyus and Z. officinale showed the highest zone (33.0 ± 0.6 mm) against standard S. aureus.
Other combinations such as A. sativum + Z. officinale and A. sativum + C. macrostachyus also had high inhibition zones (32 ± 1.2 mm).
Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)
MIC values:
Lowest MIC was 12.18 mg/mL for A. sativum against standard S. aureus.
Highest MIC was 390 mg/mL for S. incanum against P. aeruginosa.
MBC values:
Lowest MBC was 24.38 mg/mL for A. sativum against both S. aureus and E. coli.
MBC for S. incanum was greater than 390 mg/mL against K. pneumoniae and P. aeruginosa, indicating lower bactericidal potency.
Implications and Conclusions
The study demonstrated that these medicinal plants have significant potential as natural antibacterial agents for treating bacterial infections in equine wounds.
Such alternatives are particularly valuable given the growing global concern over antibiotic resistance.
The results provide a foundation for:
Future in-vivo studies to assess effectiveness and safety in live animals.
Possible development of herbal-based wound management products for veterinary use in equines.
Overall, the work supports integrating traditional medicinal plants into modern veterinary care, especially for hard-to-treat infections.
Cite This Article
APA
Beshir A, Kemal J, Abraha B, Tola EH.
(2025).
Isolation of major bacterial species associated with equine skin wounds and in-vitro antibacterial activities of selected medicinal plants.
Sci Rep, 15(1), 31942.
https://doi.org/10.1038/s41598-025-01062-7
Livestock and Fishery Office, Chiro, Oromia Regional State, Ethiopia.
Kemal, Jelalu
College of Veterinary Medicine, Haramaya University, Dire Dawa, Ethiopia.
Abraha, Bruk
College of Veterinary Medicine, Haramaya University, Dire Dawa, Ethiopia.
Tola, Eyob Hirpa
Addis Ababa University College of Veterinary Medicine and Agriculture, Bishoftu, Ethiopia. eyob.hirpa@aau.edu.et.
MeSH Terms
Animals
Equidae
Horses
Wounds and Injuries / complications
Wounds and Injuries / microbiology
Wounds and Injuries / veterinary
Bacterial Infections / drug therapy
Bacterial Infections / microbiology
Plants, Medicinal / chemistry
Plants, Medicinal / classification
Bacteria / drug effects
Bacteria / isolation & purification
Ethiopia
Staphylococcus aureus / drug effects
Staphylococcus aureus / isolation & purification
Pseudomonas aeruginosa / drug effects
Pseudomonas aeruginosa / isolation & purification
Escherichia coli / drug effects
Escherichia coli / isolation & purification
Klebsiella pneumoniae / drug effects
Klebsiella pneumoniae / isolation & purification
Plant Extracts / pharmacology
Zingiber officinale / chemistry
Garlic / chemistry
Apocynaceae / chemistry
Solanum / chemistry
Horse Diseases / drug therapy
Horse Diseases / microbiology
Conflict of Interest Statement
Declarations. Competing interests: The authors declare no competing interests.
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