Release of Hypoglycin A from Hypoglycin B and Decrease of Hypoglycin A and Methylene Cyclopropyl Glycine Concentrations in Ruminal Fluid Batch Cultures
The transformation of hypoglycin A (HGA), hypoglycin B (HGB), and methylene cyclopropyl glycine (MCPrG) in ruminal fluid batch cultures was investigated, and the effect of these toxins on the batch culture microorganisms using microbial metabolites was measured. An experiment was conducted using ovi...
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2025-01-01
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| author | Anna Maria Engel Ahmed H. El-Khatib Martin Bachmann Monika Wensch-Dorendorf Fenja Klevenhusen Stefan Weigel Robert Pieper Annette Zeyner |
| author_facet | Anna Maria Engel Ahmed H. El-Khatib Martin Bachmann Monika Wensch-Dorendorf Fenja Klevenhusen Stefan Weigel Robert Pieper Annette Zeyner |
| author_sort | Anna Maria Engel |
| collection | DOAJ |
| description | The transformation of hypoglycin A (HGA), hypoglycin B (HGB), and methylene cyclopropyl glycine (MCPrG) in ruminal fluid batch cultures was investigated, and the effect of these toxins on the batch culture microorganisms using microbial metabolites was measured. An experiment was conducted using ovine ruminal fluid batch cultures and the ANKOM RF Gas Production System over four runs, each with an incubation period of 48 h. The fermenters contained 200 mg of (i) a substrate mixture (80% cellulose, 20% starch; CSM), (ii) CSM and 1.5 mL of a solution of pure toxins (a mixture of 500 ng/mL HGA and MCPrG each; PCM), or (iii) CSM and 100 mg sycamore maple seeds (SMS). Each fermenter contained 30 mL of inoculum (ruminal fluid and buffer, 1:2 <i>v</i>/<i>v</i>). For control, autoclaved ruminal fluid was incubated with CSM, PCM, and SMS, respectively. Samples were taken from the liquid phase of the fermenters and analyzed using liquid chromatography–tandem mass spectrometry (LC/MS-MS) for sycamore maple toxins and metabolites. Microbial activity was assessed using gas production, short chain fatty acids, and NH<sub>3</sub> concentration. Additionally, pH and redox potentials were measured. In PCM, HGA and MCPrG concentrations rapidly decreased (<i>p</i> < 0.05), and were not measurable anymore after a 24 h incubation period. In SMS, the initial concentrations were 4.7 ± 1.4 µg/mL HGA, 19.9 ± 5.41 µg/mL HGB, and 1.2 ± 0.33 µg/mL MCPrG. In SMS, HGA increased in 24 h, coincidently to a decrease in HGB concentration (<i>p</i> < 0.05). We modeled a rapid conversion of HGB to HGA, accompanied by progressive HGA transformation. The concentration of MCPrG was constant until 4 h and decreased afterwards (<i>p</i> < 0.05). In SMS incubations, HGA and MCPrG concentrations of 5.6 ± 1.5 and 0.32 ± 0.090 µg/mL remained after 48 h, respectively. The HGB to HGA conversion and transformation of HGA and MCPrG also occurred in autoclaved ruminal fluid. Gas production and microbial metabolite concentrations were higher in SMS compared to CSM and PCM (<i>p</i> < 0.05), as the seeds were used as an additional substrate by the batch culture microorganisms. |
| format | Article |
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| institution | DOAJ |
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| spelling | doaj-art-a4fc87345ac14f66844e76c0cc17e1ce2025-08-20T02:45:31ZengMDPI AGToxins2072-66512025-01-011724610.3390/toxins17020046Release of Hypoglycin A from Hypoglycin B and Decrease of Hypoglycin A and Methylene Cyclopropyl Glycine Concentrations in Ruminal Fluid Batch CulturesAnna Maria Engel0Ahmed H. El-Khatib1Martin Bachmann2Monika Wensch-Dorendorf3Fenja Klevenhusen4Stefan Weigel5Robert Pieper6Annette Zeyner7German Federal Institute for Risk Assessment, 10589 Berlin, GermanyGerman Federal Institute for Risk Assessment, 10589 Berlin, GermanyGerman Federal Institute for Risk Assessment, 10589 Berlin, GermanyInstitute of Agricultural and Nutritional Sciences, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), GermanyFaculty of Organic Agriculture, University of Kassel, 37213 Witzenhausen, GermanyGerman Federal Institute for Risk Assessment, 10589 Berlin, GermanyGerman Federal Institute for Risk Assessment, 10589 Berlin, GermanyInstitute of Agricultural and Nutritional Sciences, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), GermanyThe transformation of hypoglycin A (HGA), hypoglycin B (HGB), and methylene cyclopropyl glycine (MCPrG) in ruminal fluid batch cultures was investigated, and the effect of these toxins on the batch culture microorganisms using microbial metabolites was measured. An experiment was conducted using ovine ruminal fluid batch cultures and the ANKOM RF Gas Production System over four runs, each with an incubation period of 48 h. The fermenters contained 200 mg of (i) a substrate mixture (80% cellulose, 20% starch; CSM), (ii) CSM and 1.5 mL of a solution of pure toxins (a mixture of 500 ng/mL HGA and MCPrG each; PCM), or (iii) CSM and 100 mg sycamore maple seeds (SMS). Each fermenter contained 30 mL of inoculum (ruminal fluid and buffer, 1:2 <i>v</i>/<i>v</i>). For control, autoclaved ruminal fluid was incubated with CSM, PCM, and SMS, respectively. Samples were taken from the liquid phase of the fermenters and analyzed using liquid chromatography–tandem mass spectrometry (LC/MS-MS) for sycamore maple toxins and metabolites. Microbial activity was assessed using gas production, short chain fatty acids, and NH<sub>3</sub> concentration. Additionally, pH and redox potentials were measured. In PCM, HGA and MCPrG concentrations rapidly decreased (<i>p</i> < 0.05), and were not measurable anymore after a 24 h incubation period. In SMS, the initial concentrations were 4.7 ± 1.4 µg/mL HGA, 19.9 ± 5.41 µg/mL HGB, and 1.2 ± 0.33 µg/mL MCPrG. In SMS, HGA increased in 24 h, coincidently to a decrease in HGB concentration (<i>p</i> < 0.05). We modeled a rapid conversion of HGB to HGA, accompanied by progressive HGA transformation. The concentration of MCPrG was constant until 4 h and decreased afterwards (<i>p</i> < 0.05). In SMS incubations, HGA and MCPrG concentrations of 5.6 ± 1.5 and 0.32 ± 0.090 µg/mL remained after 48 h, respectively. The HGB to HGA conversion and transformation of HGA and MCPrG also occurred in autoclaved ruminal fluid. Gas production and microbial metabolite concentrations were higher in SMS compared to CSM and PCM (<i>p</i> < 0.05), as the seeds were used as an additional substrate by the batch culture microorganisms.https://www.mdpi.com/2072-6651/17/2/46sycamore maple treesecondary plant metabolitesphytotoxinsrumen fermentationatypical myopathy |
| spellingShingle | Anna Maria Engel Ahmed H. El-Khatib Martin Bachmann Monika Wensch-Dorendorf Fenja Klevenhusen Stefan Weigel Robert Pieper Annette Zeyner Release of Hypoglycin A from Hypoglycin B and Decrease of Hypoglycin A and Methylene Cyclopropyl Glycine Concentrations in Ruminal Fluid Batch Cultures Toxins sycamore maple tree secondary plant metabolites phytotoxins rumen fermentation atypical myopathy |
| title | Release of Hypoglycin A from Hypoglycin B and Decrease of Hypoglycin A and Methylene Cyclopropyl Glycine Concentrations in Ruminal Fluid Batch Cultures |
| title_full | Release of Hypoglycin A from Hypoglycin B and Decrease of Hypoglycin A and Methylene Cyclopropyl Glycine Concentrations in Ruminal Fluid Batch Cultures |
| title_fullStr | Release of Hypoglycin A from Hypoglycin B and Decrease of Hypoglycin A and Methylene Cyclopropyl Glycine Concentrations in Ruminal Fluid Batch Cultures |
| title_full_unstemmed | Release of Hypoglycin A from Hypoglycin B and Decrease of Hypoglycin A and Methylene Cyclopropyl Glycine Concentrations in Ruminal Fluid Batch Cultures |
| title_short | Release of Hypoglycin A from Hypoglycin B and Decrease of Hypoglycin A and Methylene Cyclopropyl Glycine Concentrations in Ruminal Fluid Batch Cultures |
| title_sort | release of hypoglycin a from hypoglycin b and decrease of hypoglycin a and methylene cyclopropyl glycine concentrations in ruminal fluid batch cultures |
| topic | sycamore maple tree secondary plant metabolites phytotoxins rumen fermentation atypical myopathy |
| url | https://www.mdpi.com/2072-6651/17/2/46 |
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