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A Tandem Liquid Chromatography–Mass Spectrometry-based Approach for Metabolite Analysis of Staphylococcus aureus
Published on: March 28, 2017
1H NMR metabolic profiling of Staphylococcus pseudintermedius isolated from canine uroliths
Nahathai Uttamamul1,2, Manida Suksawat3,4, Jutarop Phetcharaburanin3,4
1Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand.
Abstract:
Staphylococcus pseudintermedius is a urease-producing bacteria which is a major cause of magnesium ammonium phosphate (MAP) urolithiasis in canine. A positive urolith culture is an important risk factor for MAP urolithiasis in canine. The mechanism underlying the metabolic changes of S. pseudintermedius after crystallization in artificial urine (AU) needs more defined baseline metabolic information. Therefore, we extensively investigated the metabolic changes of S. pseudintermedius extensively after crystallization in AU. A high urease activity and positive biofilm formation strain, entitled the S. pseudintermedius (SPMAP09) strain, was isolated from canine MAP uroliths, and analyzed using nuclear magnetic resonance (NMR) spectroscopy-based metabolomics. The molecular mechanism-specific metabolic phenotypes were clearly observed after crystallization in AU at day 3. The crystals induced by SPMAP09 were also confirmed and the major chemical composition identified as struvite. Interestingly, our findings demonstrated that a total of 11 identified metabolites were significantly changed. The levels of formate, homocarnosine, tyrosine, cis-aconitate, glycolate, ethyl malonate, valine and acetate level were significantly higher, accompanied with decreased levels of inosine, glucose, and threonine at day 3 compared with the initial time-point (day 0). In addition, our results exhibited that the glyoxylate and dicarboxylate metabolism was significantly related to the SPMAP09 strain at day 3 in AU. Thus, metabolic changes of the SPMAP09 strain after crystallization in AU potentially helps to explain the preliminary molecular mechanism for the crystals induced by S. pseudintermedius.
Insights
Staphylococcus pseudintermedius causes canine magnesium ammonium phosphate (MAP) urolithiasis. This study reveals key metabolic shifts in this bacteria after struvite crystal formation in artificial urine, offering insights into urolith development.
Area of Science:
- Veterinary Microbiology
- Bacterial Metabolism
- Urolithiasis Research
Background:
- Staphylococcus pseudintermedius is a urease-producing bacterium linked to canine magnesium ammonium phosphate (MAP) urolithiasis.
- Positive urolith culture is a significant risk factor for MAP urolithiasis in dogs.
- Understanding S. pseudintermedius metabolic changes during crystallization is crucial for elucidating urolith formation mechanisms.
Purpose of the Study:
- To investigate the metabolic alterations of S. pseudintermedius following crystallization in artificial urine (AU).
- To identify specific metabolic phenotypes and pathways involved in crystal induction by S. pseudintermedius.
Main Methods:
- Isolation and characterization of a high urease activity S. pseudintermedius strain (SPMAP09) from canine MAP uroliths.
- Analysis of metabolic changes using nuclear magnetic resonance (NMR) spectroscopy-based metabolomics.
- Confirmation of struvite crystal formation induced by the bacterial strain.
Main Results:
- Significant changes in 11 metabolites were observed in S. pseudintermedius after 3 days in AU.
- Increased levels of formate, homocarnosine, tyrosine, cis-aconitate, glycolate, ethyl malonate, valine, and acetate were noted.
- Decreased levels of inosine, glucose, and threonine were observed, alongside significant involvement of glyoxylate and dicarboxylate metabolism.
Conclusions:
- Metabolic profiling of S. pseudintermedius (SPMAP09) provides baseline data on its adaptation to crystallization conditions.
- The observed metabolic shifts, particularly in glyoxylate and dicarboxylate metabolism, are potentially linked to struvite crystal induction.
- These findings contribute to understanding the molecular mechanisms underlying S. pseudintermedius-induced canine urolithiasis.
