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Published on: July 12, 2018
Molecular Detection and Phylogenetic Analysis of Pseudomonas aeruginosa Isolated from Some Infected and Healthy
1College of Veterinary Medicine, University of Basrah, Basrah, Iraq.
Abstract:
Although P. aeruginosa is an environmental organism, it is infrequently found on the skin, mucous membranes, and in the feces of some healthy animals (wild, companion, or farm animals). P. aeruginosa produces a variety of toxins and enzymes which promote tissue invasion and damage. P. aeruginosa demonstrated resistance to several antimicrobial agents. It is of significant importance in both animal and human medicine. The present study aimed to isolate and diagnose P. aeruginosa isolates from some ruminants, cow and sheep, from different regions of Basrah, Iraq. A total of 200 samples were taken from infected and healthy ruminants, as well as the environment surrounding the animal in Basrah, Iraq. The identification of Pseudomonas aeruginosa was performed by conventional and molecular methods using the 16S rRNA gene and aroE gene by polymerase chain reaction (PCR). The recorded data pointed out that P. aeruginosa was successfully isolated from infected animals (cows and sheep) with total percentages of 46% and 22%, respectively. These percentages were obtained at 8% and 4% from healthy cows and sheep, respectively. The percentages of isolation of the environment surrounding cows and sheep were 40% and 32%, respectively. A higher percentage of infection was observed in the eye, skin, and wound swabs of cows. Healthy cows and sheep gave only three isolates of P. aeruginosa, while the environmental swabs recorded 18 isolates. Bacterial isolates were identified by culture methods and Vitek- 2. To confirm the diagnosis more accurately at the level of the species, the molecular confirmation was performed by PCR amplification of genus and species with 16S rRNA gene sequences. The results pointed out that all 10 selected isolates gave positive results, and the gene size was ≈ 1500 bp. New strains were recorded in GenBank/NCBI, and the phylogenetic tree was constructed. The isolates fall in three clads. Molecular confirmation of other isolates in this study (42 isolates) was carried out by PCR amplification of aroE gene. All PCR products of these isolates were amplified≈ 495 pb on agarose gel electrophoresis.
Insights
Pseudomonas aeruginosa was isolated from cows and sheep in Basrah, Iraq, with higher prevalence in infected animals and their environment. Molecular methods confirmed the presence of this opportunistic pathogen in ruminants and surrounding areas.
Area of Science:
- Veterinary Microbiology
- Bacterial Pathogenesis
- Molecular Diagnostics
Background:
- Pseudomonas aeruginosa is an opportunistic pathogen with significant implications in both human and veterinary medicine.
- This bacterium produces toxins and enzymes that contribute to tissue invasion and damage.
- P. aeruginosa exhibits resistance to various antimicrobial agents, posing treatment challenges.
Purpose of the Study:
- To isolate and identify Pseudomonas aeruginosa from ruminants (cows and sheep) and their environment in Basrah, Iraq.
- To characterize the prevalence of P. aeruginosa in infected and healthy animals and their surroundings.
- To confirm the identification using conventional and molecular techniques.
Main Methods:
- Collection of 200 samples from cows, sheep, and their environment in Basrah, Iraq.
- Isolation and identification using conventional culture methods and Vitek-2 system.
- Molecular confirmation via Polymerase Chain Reaction (PCR) targeting the 16S rRNA and aroE genes.
Main Results:
- P. aeruginosa was isolated from 46% of infected cows and 22% of infected sheep.
- Isolation rates from healthy cows and sheep were 8% and 4%, respectively.
- Environmental samples showed isolation rates of 40% for cow surroundings and 32% for sheep surroundings, with higher infection rates in eye, skin, and wound swabs of cows.
Conclusions:
- Pseudomonas aeruginosa is prevalent in ruminant populations and their environment in Basrah, Iraq.
- Molecular methods, including 16S rRNA and aroE gene PCR, effectively confirmed the species identification.
- The study identified new strains and established a phylogenetic relationship among the isolates.
