Research article · 2023 · Full text
International Journal of Veterinary Sciences and Animal HusbandryDetection of virulence factors of Pseudomonas aeruginosa from dog’s wound infections by using biochemical and molecular methods
Volume 8(3) · Pages 01–07 · DOI: 10.22271/veterinary.2023.v8.i3a.506 ↗
Research areasAntimicrobial Peptides and Activities
Abstract
Author-writtenThis investigation's goal was to find Pseudomonas aeruginosa virulence factors in canine wound infections. 100 samples were gathered and tested for eight virulence variables utilizing biochemical and molecular techniques for this purpose. According to the findings, women (72%) had the highest isolation ratio. The virulence factors were found in various ratios, with capsule being found in (16.6%) of them, amylase enzyme detected in (3.3%), hemolysin enzyme detected in (70%), protease detected in (10%), phospholipase enzyme detected in (83.3%), urease enzyme detected in (100%), DNAase enzyme detected in (80%), pigments production detected in (93.3%) from wound infections isolates, gal and lasA (86.6%), toxA (16.6%), pslA (96.6%),16SrRNA (100%). From this research, we conclude that dogs are more prone to bacterial infections.
Summary
Source-checkedPlain-language reading of the full paper, prepared with AI and checked against the text. Each point links to the authors' own words.
Why it matters
- Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium capable of infecting both humans and animals, causing various conditions such as pneumonia and dermatitis in animals. View evidence ↗
- This bacterium is noted for its high resistance to common antibacterial agents, including tetracyclines and penicillins. View evidence ↗
- The pathogen utilizes various virulence factors, such as phospholipases and hemolysin, to degrade the host's immune system effector molecules. View evidence ↗
The question
- The study aimed to identify the specific virulence factors of Pseudomonas aeruginosa present in wound infections of dogs. View evidence ↗
What they did
- The research analyzed 100 dog wound infection samples, consisting of 72 females and 28 males. View evidence ↗
- Bacterial isolation involved culturing swabs on nutrient, blood, and MacConkey agar at 37°C for 24 hours, followed by subculturing on Pseudomonas chromogenic agar. View evidence ↗
- Identification of the bacteria was performed using biochemical tests, including catalase, oxidase, and urease tests, as well as growth checks on nutrient agar at 42°C. View evidence ↗
What they found
- The highest isolation rate of the bacteria was found in female dogs (72%). View evidence ↗
- Urease enzyme was detected in 100% (30/30) of the Pseudomonas isolates. View evidence ↗
- Molecular analysis showed that the 16SrRNA gene was present in 100% (30/30) of the isolates. View evidence ↗
What it means
- The authors conclude that dogs are highly susceptible to bacterial infections. View evidence ↗
Visual summary
2 visualsIllustrations explain the paper; charts are replotted from the paper's own table values.
Study Workflow
AI-generated conceptual illustration (Pseudomonas aeruginosa bacteria from dog wound swabs). It explains the idea and does not show study data.
View source evidence ↗
%: gal vs lasA vs toxA vs pslA vs 16SrRNA
Replotted from Table 2 (page 2) of the original paper. Every bar is the paper's own table value.
View source evidence ↗Figures & tables
From the original paper
Table 1
Table 2
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Figure 13Introduction
A saprophytic and opportunistic pathogen, Pseudomonas aeruginosa is a G-bacterium that may infect both people and animals [2] . It causes pneumonia, dermatitis, mastitis, metritis, enteritis, and dermatitis in animals [16] . Despite the fact that Pseudomonas aeruginosa organisms have been found to be highly resistant to typical antibacterial substances like penicillin's and tetracyclines [19] . Pseudomonas aeruginosa has a variety of virulence factors, including hemolysin, proteases, ureases, and phospholipases, which break down immune system effector molecules. They also have components necessary for the cellular and tissue structure of the host [12,21] . Pseudomonas aeruginosa has a solo polar flagellum that it uses to move, and by adhering to the asialyated glycolipid, it can attach to host epithelial cells [5] . Exotoxin A, which is present in both pathogenic and nonpathogenic strains of Pseudomonas aeruginosa and is located in its outer membrane proteins (toxA), prevents protein synthesis by ADP ribosylation of elongation factor 2 and consequently causes cell death [10] . (pslA): This gene was discovered in 517 genera and is a cluster that participates in cell-cell and/or cell-surface interaction in the formation of biofilms [11,22] . The Pseudomonas aeruginosa gene (lasA), which is implicated in both proteolysis and elastolysis, was the first to be cloned and sequenced. Using genetic methods, its importance and function have been investigated [20] . (gal): cytoplasmic localization, present in both pathogenic and nonpathogenic strains 642 genera possessed this gene [20] . (16SrRNA): The majority of housekeeping genetic markers have been utilized to research bacterial phylogeny and taxonomy using 16S rRNA gene sequences for a variety of reasons [24] . These proteins may be a reliable factor for Pseudomonas aeruginosa fast identification in clinical samples because they are only present in this bacterium [7] . Consequently, the objectives of this study were to find Pseudomonas aeruginosa virulence factors in canine wound infections.
Materials and methods
Baghdad province served as the design hub for this study. The samples were collected from February to December 2022 from (Al-Shifa veterinary clinic, Mila veterinary clinic, dr. Shefa badrana clinic, Adhamiya veterinary clinic, Shabaad veterinary clinic\in the Adhamiya area), (Al-Farah veterinary clinic, dr. Muhannad veterinary clinic\ Al-Bayaa 20 th Street). This study included a total of 100 (28 male and 72 female) of dog's wound infection. The samples were obtained and delivered right away to the labs of Baghdad University and Al-Nahrain University for bacterial culturing and molecular technique.
Isolation of Pseudomonas aeruginosa from dog's wound infection
After transporting to the laboratory, the swabs were cultured on MacConkey agar, blood agar, nutrient agar and incubated at 37°C for 24 hours. After incubation, were subcultured on Pseudomonas chromogenic agar.
Identification of Pseudomonas aeruginosa A. Colonial morphology
The form, color, diameter, and odor of the colonies of the bacterial isolates that grow on blood agar, MacConkey agar, nutritional agar, and Pseudomonas chromogenic agar were characterized [6] .
B. Microscopical examination
One isolated colony was moved to a microscope slide, which was then properly treated and stained with Gram stain. Gram reaction, cell arrangement, and cell morphology were noted [9] .
C. Biochemical tests
Oxidase test, catalase test, urease test, Lactose fermentation on MacConkey, Growth on Nutrient agar at 42 ˚C [8] .
Results
The ratio of virulence factors were capsule: 16.6%, amylase enzyme: 3.3% (figure 6) , hemolysin enzyme: 70% (figure 2), protease: 10% (figure 3), phospholipase enzyme: 83.3% (figure 7), urease enzyme: 100% (figure 5), DNAase enzyme: 80% (figure 4), pigments: 93.3%. Table 1 & figure 13 demonstrated Pseudomonas aeruginosa virulence components that were identified from canine wound infections. Lane L:molecular ladder of 100bp. Electrophorasis was carried out in 1.5% agaros gel supplied with Ethidium Bromide at 7V/cm for 90 minutes. Lane L:molecular ladder of 100bp . Electrophorasis was carried out in 1.5% agaros gel supplied with Ethidium Bromide at 7V/cm for 90 minutes. Lane L: molecular ladder of 100bp. Electrophorasis was carried out in 1.5% agaros gel supplied with ethidium bromide at 7V/cm for 90 minutes. Lane L: molecular ladder of 100bp . Electrophorasis was carried out in 1.5% agaros gel supplied with Ethidium Bromide at 7V/cm for 90 minutes.
| Samples of dog’s wound infections (100) | — | — | — |
|---|---|---|---|
| Pseudomonas isolates (30) | |||
| Virulence factors | No | % | |
| 1 | Capsule | 5 | 16.6% |
| 2 | Amylase enzyme | 1 | 3.3% |
| 3 | Hemolysin enzyme | 21 | 70% |
| 4 | Protease | 3 | 10% |
| 5 | Phospholipase enzyme | 25 | 83.3% |
| 6 | Urease enzyme | 30 | 100% |
| 7 | DNAase enzyme | 24 | 80% |
| 8 | Pigments | 28 | 93.3% |
Table text extracted from the original PDF; superscript letters are shown inline. The original table image is below.
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Discussion
In this investigation, P. aeruginosa was isolated from dog wound infections. Dog samples had the highest isolation rate, which was consistent with [15] . The isolation of P. aeruginosa from dog's wound infections, all the isolates have the ability to grow at 42C• but not at 4C• this result agreement [4] . Some isolates appeared to have capsule, this result differ from result showed by [1] were 0%, while [3] reported a 42.8% success rate. To protect bacteria from other types of immunological invasion, capsules play a significant role in preventing phagocytosis [17] . The detection of the hemolysin enzyme was 70%, in agreement with [15,1] and differing from (5were 100%). 80% of samples tested positive for DNAase, which is a different outcome from the 15% who tested negative. The majority of isolates produced pigments when tested, which was in consistence with [13] . The virulence of P. aeruginosa is significantly influenced by pigments, which also lower host immunity [18] . As urease is an enzyme that is present in all isolates, it is able to divert urea into CO2 and NH3 and raise PH, all of which promote bacterial growth. This result varies from that reported by [15,5] were negative. Also, the study's results for the protease detection test were favorable. By destroying host cells and impairing immune defenses like skin and mucous membranes, protease enzymes promote infection [14, 15] . In current study (gal, toxA, lasA, PslA, 16SrRNA) genes were detected in most isolates. The toxA gene was found in 16.6% of P. aeruginosa isolated from wound infections in dogs. ToxA is a naturally occurring genetic segment on the P. aeruginosa chromosome that controls the generation of exotoxin A, this study differs from what was reported [4,23,25,26] .
Figures and tables from the paper
| Samples of dog’s wound infections (100) | — | — | — |
|---|---|---|---|
| Pseudomonas isolates (30) | |||
| Virulence factors | N0 | % | |
| 1 | gal | 26 | 86.6% |
| 2 | lasA | 26 | 86.6% |
| 3 | toxA | 5 | 16.6% |
| 4 | pslA | 29 | 96.6% |
| 5 | 16SrRNA | 30 | 100% |
Table text extracted from the original PDF; superscript letters are shown inline. The original table image is below.
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References
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- Outbreak of Carbapenem-Resistant High-Risk Clone ST244 of Pseudomonas aeruginosa in Dogs and Cats in Algeria ↗
Amina Badis, Nouzha Heleili, Manel Merradi, Ammar Ayachi, Piera Anna Martino, Gabriele Meroni et al. · Antibiotics · 2025
Background/Objectives: Pseudomonas aeruginosa causes chronic infections in humans and animals, especially cats and dogs. This bacterium’s ability to adapt and acquire antibiotic resistance traits may complicate and exacerbate antibacterial therapy. This study aimed to evaluate the antibiotic resistance patterns, virulence factors and ability to form biofilms of P. aeruginosa strains isolated from Algerian dogs and cats. Methods: Nineteen samples were…
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