Identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profiling
Objective Cow urine possesses several bioactive properties but the responsible components behind these bioactivities are still far from identified. In our study, we tried to identify the possible components behind the antimicrobial activity of cow urine by exploring the peptidome and metabolome. Met...
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| Format: | Article |
| Language: | English |
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Asian-Australasian Association of Animal Production Societies
2023-07-01
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| Series: | Animal Bioscience |
| Subjects: | |
| Online Access: | http://www.animbiosci.org/upload/pdf/ab-22-0249.pdf |
| _version_ | 1797787275511201792 |
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| author | Rohit Kumar Jai Kumar Kaushik Ashok Kumar Mohanty Sudarshan Kumar |
| author_facet | Rohit Kumar Jai Kumar Kaushik Ashok Kumar Mohanty Sudarshan Kumar |
| author_sort | Rohit Kumar |
| collection | DOAJ |
| description | Objective Cow urine possesses several bioactive properties but the responsible components behind these bioactivities are still far from identified. In our study, we tried to identify the possible components behind the antimicrobial activity of cow urine by exploring the peptidome and metabolome. Methods We extracted peptides from the urine of Sahiwal cows belonging to three different physiological states viz heifer, lactation, and pregnant, each group consisting of 10 different animals. The peptides were extracted using the solid phase extraction technique followed by further extraction using ethyl acetate. The antimicrobial activity of the aqueous extract was evaluated against different pathogenic strains like Staphylococcus aureus, Escherichia coli, and Streptococcus agalactiae. The safety of urinary aqueous extract was evaluated by hemolysis and cytotoxicity assay on the BuMEC cell line. The urinary peptides were further fractionated using high-performance liquid chromatography (HPLC) to identify the fraction(s) containing the antimicrobial activity. The HPLC fractions and ethyl acetate extract were analyzed using nLC-MS/MS for the identification of the peptides and metabolites. Results A total of three fractions were identified with antimicrobial activity, and nLC-MS/MS analysis of fractions resulted in the identification of 511 sequences. While 46 compounds were identified in the metabolite profiling of organic extract. The urinary aqueous extract showed significant activity against E. coli as compared to S. aureus and S. agalactiae and was relatively safe against mammalian cells. Conclusion The antimicrobial activity of cow urine is a consequence of the feeding habit. The metabolites of plant origin with several bioactivities are eliminated through urine and are responsible for their antimicrobial nature. Secondly, the plethora of peptides generated from the activity of endogenous proteases on protein shed from different parts of tissues also find their way to urine. Some of these sequences possess antimicrobial activity due to their amino acid composition. |
| first_indexed | 2024-03-13T01:19:41Z |
| format | Article |
| id | doaj.art-a424391a5c5a4823a9d630209baab4b0 |
| institution | Directory Open Access Journal |
| issn | 2765-0189 2765-0235 |
| language | English |
| last_indexed | 2024-03-13T01:19:41Z |
| publishDate | 2023-07-01 |
| publisher | Asian-Australasian Association of Animal Production Societies |
| record_format | Article |
| series | Animal Bioscience |
| spelling | doaj.art-a424391a5c5a4823a9d630209baab4b02023-07-05T04:51:53ZengAsian-Australasian Association of Animal Production SocietiesAnimal Bioscience2765-01892765-02352023-07-013671130114210.5713/ab.22.024925004Identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profilingRohit Kumar0Jai Kumar Kaushik1Ashok Kumar Mohanty2Sudarshan Kumar3 ICAR-National Dairy Research Institute, Cell Biology and Proteomics Lab, Animal Biotechnology Center (ABTC), Karnal, Haryana, 132001, India ICAR-National Dairy Research Institute, Cell Biology and Proteomics Lab, Animal Biotechnology Center (ABTC), Karnal, Haryana, 132001, India ICAR-National Dairy Research Institute, Cell Biology and Proteomics Lab, Animal Biotechnology Center (ABTC), Karnal, Haryana, 132001, India ICAR-National Dairy Research Institute, Cell Biology and Proteomics Lab, Animal Biotechnology Center (ABTC), Karnal, Haryana, 132001, IndiaObjective Cow urine possesses several bioactive properties but the responsible components behind these bioactivities are still far from identified. In our study, we tried to identify the possible components behind the antimicrobial activity of cow urine by exploring the peptidome and metabolome. Methods We extracted peptides from the urine of Sahiwal cows belonging to three different physiological states viz heifer, lactation, and pregnant, each group consisting of 10 different animals. The peptides were extracted using the solid phase extraction technique followed by further extraction using ethyl acetate. The antimicrobial activity of the aqueous extract was evaluated against different pathogenic strains like Staphylococcus aureus, Escherichia coli, and Streptococcus agalactiae. The safety of urinary aqueous extract was evaluated by hemolysis and cytotoxicity assay on the BuMEC cell line. The urinary peptides were further fractionated using high-performance liquid chromatography (HPLC) to identify the fraction(s) containing the antimicrobial activity. The HPLC fractions and ethyl acetate extract were analyzed using nLC-MS/MS for the identification of the peptides and metabolites. Results A total of three fractions were identified with antimicrobial activity, and nLC-MS/MS analysis of fractions resulted in the identification of 511 sequences. While 46 compounds were identified in the metabolite profiling of organic extract. The urinary aqueous extract showed significant activity against E. coli as compared to S. aureus and S. agalactiae and was relatively safe against mammalian cells. Conclusion The antimicrobial activity of cow urine is a consequence of the feeding habit. The metabolites of plant origin with several bioactivities are eliminated through urine and are responsible for their antimicrobial nature. Secondly, the plethora of peptides generated from the activity of endogenous proteases on protein shed from different parts of tissues also find their way to urine. Some of these sequences possess antimicrobial activity due to their amino acid composition.http://www.animbiosci.org/upload/pdf/ab-22-0249.pdfantimicrobial peptidescow urinemass spectrometrymetabolitessolid-phase extraction |
| spellingShingle | Rohit Kumar Jai Kumar Kaushik Ashok Kumar Mohanty Sudarshan Kumar Identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profiling Animal Bioscience antimicrobial peptides cow urine mass spectrometry metabolites solid-phase extraction |
| title | Identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profiling |
| title_full | Identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profiling |
| title_fullStr | Identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profiling |
| title_full_unstemmed | Identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profiling |
| title_short | Identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profiling |
| title_sort | identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profiling |
| topic | antimicrobial peptides cow urine mass spectrometry metabolites solid-phase extraction |
| url | http://www.animbiosci.org/upload/pdf/ab-22-0249.pdf |
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