Ligilactobacillus agilis W70: a probiotic with capacity to enhance ammonia assimilation in the rumen

Ruminants excrete both undigested and excess consumed nitrogen (N) through feces and urine, which results in the waste of feed protein and environmental pollution. This study aims to screen bacteria to identify strains with potential to improve nitrogen utilization in the rumen. In a tube screening...

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Main Authors: Xiaowei Duan, Stafford Vigors, Rui Ma, Lu Ma, Jingang Gu, Dengpan Bu
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-01-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2024.1498109/full
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author Xiaowei Duan
Xiaowei Duan
Stafford Vigors
Rui Ma
Lu Ma
Lu Ma
Jingang Gu
Dengpan Bu
author_facet Xiaowei Duan
Xiaowei Duan
Stafford Vigors
Rui Ma
Lu Ma
Lu Ma
Jingang Gu
Dengpan Bu
author_sort Xiaowei Duan
collection DOAJ
description Ruminants excrete both undigested and excess consumed nitrogen (N) through feces and urine, which results in the waste of feed protein and environmental pollution. This study aims to screen bacteria to identify strains with potential to improve nitrogen utilization in the rumen. In a tube screening test, when 30 mmol/L ammonium sulfate was added as the only nitrogen source in liquid medium, among 115 bacterial strains belonging to Bacillus and lactic-acid bacteria, 10 strains with the highest growth (OD600nm level) in comparison to the other evaluated strains were identified, and of these, Ligilactobacillus agilis W70 and Limosilactobacillus fermentum M50, had the highest ammonia nitrogen utilizing capabilities of ammonium sulfate (32.09 and 40.64%, respectively). A subsequent in vitro fermentation experiment was conducted with the diet consisting of 0.5 g TMR, 50 mL buffer solution, and 25 mL fresh rumen fluid per serum bottle which was incubated at 39°C for 24 h. The experiment consisted of 3 treatments, CTL, L. agilis W70 (1 × 109 cfu/mL), and L. fermentum M50 (1 × 109 cfu/mL), each treatment had 6 replicates, repeated across 3 batches. The addition of L. agilis W70 decreased NH3N (p < 0.01), the ratio of acetate to propionate (A:P) (p < 0.01), and increased the yields of microbial protein (MCP) (p < 0.01) and enzymatic activities of glutamate dehydrogenase (GDH) (p < 0.05) and glutamine synthetase (GS) (p < 0.05). Further genome analysis revealed that the L. agilis W70 harbors the glutamate dehydrogenase and glutamine synthetase-encoding genes gdhA and glnA that play key roles in ammonia utilization. This study identified L. agilis W70 as a strain which exhibits high NH3N utilization capability and enhances ammonia assimilation in vitro. The strain was further characterized to elucidate this ammonia assimilation potential. Further studies will be conducted to develop strain L. agilis W70 as a new feed additive to improve the nitrogen utilization efficiency in ruminant animals.
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publisher Frontiers Media S.A.
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series Frontiers in Microbiology
spelling doaj-art-2c68bd4105d548b9b0d36a573e5991d62025-01-17T06:50:59ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-01-011510.3389/fmicb.2024.14981091498109Ligilactobacillus agilis W70: a probiotic with capacity to enhance ammonia assimilation in the rumenXiaowei Duan0Xiaowei Duan1Stafford Vigors2Rui Ma3Lu Ma4Lu Ma5Jingang Gu6Dengpan Bu7State Key Laboratory of Animal Nutrition and Feeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, ChinaSchool of Agriculture and Food Science, University College Dublin, Dublin, IrelandSchool of Agriculture and Food Science, University College Dublin, Dublin, IrelandInstitute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing, ChinaState Key Laboratory of Animal Nutrition and Feeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, ChinaNational Center of Technology Innovation for Dairy, Inner Mongolia, ChinaInstitute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing, ChinaState Key Laboratory of Animal Nutrition and Feeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, ChinaRuminants excrete both undigested and excess consumed nitrogen (N) through feces and urine, which results in the waste of feed protein and environmental pollution. This study aims to screen bacteria to identify strains with potential to improve nitrogen utilization in the rumen. In a tube screening test, when 30 mmol/L ammonium sulfate was added as the only nitrogen source in liquid medium, among 115 bacterial strains belonging to Bacillus and lactic-acid bacteria, 10 strains with the highest growth (OD600nm level) in comparison to the other evaluated strains were identified, and of these, Ligilactobacillus agilis W70 and Limosilactobacillus fermentum M50, had the highest ammonia nitrogen utilizing capabilities of ammonium sulfate (32.09 and 40.64%, respectively). A subsequent in vitro fermentation experiment was conducted with the diet consisting of 0.5 g TMR, 50 mL buffer solution, and 25 mL fresh rumen fluid per serum bottle which was incubated at 39°C for 24 h. The experiment consisted of 3 treatments, CTL, L. agilis W70 (1 × 109 cfu/mL), and L. fermentum M50 (1 × 109 cfu/mL), each treatment had 6 replicates, repeated across 3 batches. The addition of L. agilis W70 decreased NH3N (p < 0.01), the ratio of acetate to propionate (A:P) (p < 0.01), and increased the yields of microbial protein (MCP) (p < 0.01) and enzymatic activities of glutamate dehydrogenase (GDH) (p < 0.05) and glutamine synthetase (GS) (p < 0.05). Further genome analysis revealed that the L. agilis W70 harbors the glutamate dehydrogenase and glutamine synthetase-encoding genes gdhA and glnA that play key roles in ammonia utilization. This study identified L. agilis W70 as a strain which exhibits high NH3N utilization capability and enhances ammonia assimilation in vitro. The strain was further characterized to elucidate this ammonia assimilation potential. Further studies will be conducted to develop strain L. agilis W70 as a new feed additive to improve the nitrogen utilization efficiency in ruminant animals.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1498109/fullLigilactobacillus agilisNH3Nrumen fermentationscreeningfunctionalphysicochemical characterization
spellingShingle Xiaowei Duan
Xiaowei Duan
Stafford Vigors
Rui Ma
Lu Ma
Lu Ma
Jingang Gu
Dengpan Bu
Ligilactobacillus agilis W70: a probiotic with capacity to enhance ammonia assimilation in the rumen
Frontiers in Microbiology
Ligilactobacillus agilis
NH3N
rumen fermentation
screening
functional
physicochemical characterization
title Ligilactobacillus agilis W70: a probiotic with capacity to enhance ammonia assimilation in the rumen
title_full Ligilactobacillus agilis W70: a probiotic with capacity to enhance ammonia assimilation in the rumen
title_fullStr Ligilactobacillus agilis W70: a probiotic with capacity to enhance ammonia assimilation in the rumen
title_full_unstemmed Ligilactobacillus agilis W70: a probiotic with capacity to enhance ammonia assimilation in the rumen
title_short Ligilactobacillus agilis W70: a probiotic with capacity to enhance ammonia assimilation in the rumen
title_sort ligilactobacillus agilis w70 a probiotic with capacity to enhance ammonia assimilation in the rumen
topic Ligilactobacillus agilis
NH3N
rumen fermentation
screening
functional
physicochemical characterization
url https://www.frontiersin.org/articles/10.3389/fmicb.2024.1498109/full
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