CoChat
Sign Up
Cited by 68Ruminant Nutrition and Digestive PhysiologyProbiotics and Fermented FoodsAnimal Nutrition and Physiology

Potential of Lactic Acid Bacteria Isolated From Different Forages as Silage Inoculants for Improving Fermentation Quality and Aerobic Stability

Open access
Puntillo, Melisa, Gaggiotti, Mónica, Oteiza, Juan Martín, Binetti, Ana, Massera, Ariel, Vinderola, Gabriel·2020·Frontiers in Microbiology
DOI 10.3389/fmicb.2020.586716PMC PMC7752947
Open this paper in CoChat — free accountOpen PDF

Abstract

We aimed at isolating lactic acid bacteria (LAB) from different plant materials to study their crossed-fermentation capacity in silos and to find strains able to confer enhanced aerobic stability to silage. A total of 129 LAB isolates were obtained from lucerne (alfalfa), maize, sorghum, ryegrass, rice, barley, canola, Gatton panic, Melilotus albus, soy, white clover, wheat, sunflower, oat, and moha. Four Lactiplantibacillus plantarum subsp. plantarum strains (isolated from oat, lucerne, sorghum, or maize) were selected for their growth capacity. Identity (16S sequencing) and diversity (RAPD-PCR) were confirmed. Fermentative capacity (inoculated at 10 4, 10 5, 10 6, 10 7 CFU/g) was studied in maize silage and their cross-fermentation capacity was assessed in oat, lucerne, sorghum, and maize. Heterofermentative strains with the highest acetic acid production capacity conferred higher aerobic stability to maize silages. Regardless the source of isolation, L. plantarum strains, inoculated at a rate of 10 6 CFU/g, were effective to produce silage from different plant materials. From more than 100 isolates obtained, the application of a succession of experiments allowed us to narrow down the number of potential candidates of silage inoculants to two strains. Based on the studies made, L. plantarum LpM15 and Limosilactobacillus fermentum LfM1 showed potential to be used as inoculants, however further studies are needed to determine their performance when inoculated together. The former because it positively influenced different quality parameters in oat, lucerne, sorghum, and maize silage, and the latter because of its capacity to confer enhanced aerobic stability to maize silage. The rest of the strains constitute a valuable collection of autochthonous strains that will be further studied in the future for new applications in animal or human foods.

AI snapshot

Written from the paper’s full text.

TL;DR

A total of 129 LAB isolates from 15 forage crops were screened, and four *Lactiplantibacillus plantarum* strains were tested as silage inoculants, with LpM15 improving quality parameters across oat, lucerne, sorghum, and maize silages. The heterofermentative *Limosilactobacillus fermentum* LfM1 conferred enhanced aerobic stability to maize silage, and the authors propose LpM15 and LfM1 as promising inoculant candidates pending further study.

Key findings

  • From 15 plant materials, 161 presumptive LAB isolates were obtained from 14 crops (not canola), and 129 were identified by MALDI-TOF; *L. pentosus/plantarum* was the most frequent homofermentative group, while *L. brevis* was the most frequent heterofermentative species, found in 6 of 14 plant materials.
  • Four *L. plantarum* strains (LpAv, LpA3, LpS13, LpM15) were selected; at 10⁶ CFU/g they were effective across forages, and LpM15 reduced oat silage ADF and NDF to 33.37 ± 1.39% and 52.20 ± 2.02%, respectively, versus control values of 35.53 ± 0.47% and 55.18 ± 0.40%.
  • The highest acetic acid producers among heterofermentative isolates were *L. fermentum* LfM1 (7.56 ± 0.73 g/l), *L. brevis* LbM6 (6.89 ± 0.43 g/l), *L. brevis* LbB2 (6.39 ± 0.58 g/l), and *L. fermentum* LfSY1 (5.87 ± 0.25 g/l); *L. buchneri* reference strains Ls141 and 463 produced 5.42 ± 0.36 g/l and 5.30 ± 0.53 g/l.
  • In maize silage after 90 days, control lost aerobic stability at 123 h (5 days), LbB2 at 212 h (8 days), and LfSY1 at 224 h (9 days), whereas Ls141, 463, LfM1, and LbM6 kept silos stable for at least 12 days, when the experiment was stopped.

Methods

  • Samples from 15 forage crops were collected in duplicate from Santa Fe and Buenos Aires provinces, Argentina, during 2016; LAB were isolated on MRS agar, identified by MALDI-TOF MS, and selected *L. plantarum* strains were confirmed by 16S rRNA sequencing and RAPD-PCR.
  • Lab-scale micro-silos were used: maize silage dose-response at 10⁴, 10⁵, 10⁶, and 10⁷ CFU/g; crossed-fermentation of four *L. plantarum* strains in oat, lucerne, sorghum, and maize at 10⁶ CFU/g; microbial counts, pH, and chemical composition were measured over fermentation.
  • Heterofermentative strains were screened for lactic and acetic acid production by HPLC, then selected strains were inoculated at 10⁶ CFU/g in maize silage and aerobic stability was measured after 90 days as time for internal silage temperature to rise 2°C above room temperature; data were analyzed by one-way ANOVA with Tukey or Dunnett tests (p < 0.05).

Limitations

  • The paper states that further studies are needed to determine the performance of LpM15 and LfM1 when inoculated together, and that the proposed metabolic adaptation at the lowest inoculation rate is a hypothesis yet to be proved.
  • Evident from the text: the work used lab-scale micro-silos and in vitro forage-based media rather than farm-scale silos, and samples came from one collection year and region, so broader ensiling and animal-performance outcomes are not established.
  • Evident from the text: the aerobic stability assay was stopped at 12 days, so the maximum stability duration of the most effective strains was not determined.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.3389/fmicb.2020.586716

Cites

59 of 59 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 52 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Except from Canola, a total of 161 presumptive LAB isolates were obtained from the other 14 forage crops.

    “Except from Canola, a total of 161 presumptive LAB isolates were obtained from the other 14 forage crops.”
    Quote in the paperNot a substantial claim
  • Findingown data

    From more than 100 isolates obtained, the application of a succession of experiments allowed us to narrow down the number of potential candidates of silage inoculants to two strains.

    “From more than 100 isolates obtained, the application of a succession of experiments allowed us to narrow down the number of potential candidates of silage inoculants to two strains.”
    • Also stated in: § Conclusion · plain
      “From more than 100 isolates obtained, the application of a succession of experiments allowed us to narrow down the number of potential candidates of silage inoculants to two strains.”
    Quote in the paperSubstantial claim
  • Findingown data

    L. plantarum LpM15 positively influenced different quality parameters in oat, lucerne, sorghum, and maize silage.

    “The former because it positively influenced different quality parameters in oat, lucerne, sorghum, and maize silage”
    Quote in the paperSubstantial claim
  • Findingown data

    A total of 129 LAB isolates were obtained from lucerne (alfalfa), maize, sorghum, ryegrass, rice, barley, canola, Gatton panic, Melilotus albus, soy, white clover, wheat, sunflower, oat, and moha.

    “A total of 129 LAB isolates were obtained from lucerne (alfalfa), maize, sorghum, ryegrass, rice, barley, canola, Gatton panic, Melilotus albus, soy, white clover, wheat, sunflower, oat, and moha.”
    Quote in the paperSubstantial claim
  • Findingnone given

    Heterofermentative strains with the highest acetic acid production capacity conferred higher aerobic stability to maize silages.

    “Heterofermentative strains with the highest acetic acid production capacity conferred higher aerobic stability to maize silages.”
    Quote in the paperSubstantial claim
  • Classified as probably not a substantial claim. 30
  • The quote only partly states this claim. 2
  • May combine two claims. 36

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    Fifty-one samples from 15 forage crops were processed.

    “Fifty-one samples from 15 forage crops were processed.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Findingnone given

    Four Lactiplantibacillus plantarum subsp. plantarum strains (isolated from oat, lucerne, sorghum, or maize) were selected for their growth capacity.

    “Four Lactiplantibacillus plantarum subsp. plantarum strains (isolated from oat, lucerne, sorghum, or maize) were selected for their growth capacity.”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    Limosilactobacillus fermentum LfM1 has the capacity to confer enhanced aerobic stability to maize silage.

    “the latter because of its capacity to confer enhanced aerobic stability to maize silage”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In terms of cell counts, the higher growth was supported by ABM (>1.6 Δlog10 CFU/ml), followed by MBM (1.2-1.4 Δlog10 CFU/ml).

    “In terms of cell counts, the higher growth was supported by ABM (>1.6 Δlog10 CFU/ml), followed by MBM (1.2–1.4 Δlog10 CFU/ml)”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    All heterofermentative strains under study were effective in conferring enhanced aerobic stability, with different capacity among them.

    “In general, all strains under study were effective in conferring enhanced aerobic stability, with different capacity among them.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Similar growth kinetics were observed for the four strains on each plant material, but different cell counts were observed after 24 and 48 h of growth in each plant material.

    “Similar growth kinetics (figures not shown) were observed for the four strains on each plant material, but different cell counts were observed after 24 and 48 h of growth in each plant material (Figure 3).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In sorghum silage after 30 days, inoculated silages displayed pH lower than 3.84, while control pH was above 4.

    “Furthermore, after 30 days of fermentation, inoculated silages displayed a pH lower than 3.84, while the pH of the control was above 4.”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    For the dose of 105 CFU/g in maize silage, pH differences were observed among all L. plantarum strains (p < 0.05).

    “For the dose of 105 CFU/g, differences were observed among all L. plantarum strains (p < 0.05).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In sorghum silage after 24 h, pH values were 4.20 ± 0.01 for LpAv, 4.29 ± 0.05 for LpA3, 4.26 ± 0.13 for LpS13, 4.16 ± 0.02 for LpM15, and 4.71 ± 0.04 for control.

    “After 24 h the pH values were 4.20 ± 0.01 (LpAv, p < 0.001), 4.29 ± 0.05 (LpA3, p < 0.001), 4.26 ± 0.13 (LpS13, p < 0.001), 4.16 ± 0.02 (LpM15, p = 0.000) and 4.71 ± 0.04 (control).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    L. plantarum strains obtained from maize reduced pH values of the MBM from 6.19 to 3.91-4.25 after 24 h, and then a negligible additional pH drop after 48 h of incubation was observed.

    “L. plantarum strains obtained from maize reduced pH values of the MBM from 6.19 to 3.91–4.25 after 24 h, and then a negligible additional pH drop after 48 h of incubation was observed.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The concentrations of acetic acid produced by L. buchneri Ls141 and L. buchneri 463 were 5.42 ± 0.36 g/l and 5.30 ± 0.53 g/l, respectively.

    “The concentrations of acetic acid produced by L. buchneri Ls141 and L. buchneri 463 were 5.42 ± 0.36 g/l and 5.30 ± 0.53 g/l, respectively.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Most isolates grew from 1 to 1.5 log orders, while some isolates obtained from maize were able to reach the expected 2 log orders of growth in MBM.

    “Most isolates grew from 1 to 1.5 log orders, while some isolates obtained from maize were able to reach the expected 2 log orders of growth in MBM (as 1% v/v inoculum was used).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In maize silage after 30 days, pH values were lower than 3.90 in all cases, and LpS13 silage was the only one with a significantly lower pH value compared to control.

    “After 30 days of fermentation, pH values were lower than 3.90 in all cases, being the LpS13 silage the only one with a significantly lower pH value, compared to control.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    LpS13 displayed the highest growth capacity whereas the growth of LpAv was significantly lower in the four media (p < 0.05), compared to LpS13, particularly in SBM.

    “LpS13 displayed the highest growth capacity whereas the growth of LpAv was significantly lower in the four media (p < 0.05), compared to LpS13, particularly in SBM.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Findingown data

    For the highest dose assessed in maize silage, pH differences were observed between LpM15 and LpAv (p = 0.002) and LpA3 (p = 0.006).

    “Finally, for the highest dose assessed, differences were observed between LpM15 and LpAv (p = 0.002) and LpA3 (p = 0.006).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    After 90 days of fermentation, DM ranged from 38.9 ± 0.2% to 43.1 ± 3.4%, with no significant differences among treatments (p > 0.05).

    “After 90 days of fermentation, DM ranged from 38.9 ± 0.2% to 43.1 ± 3.4%, but again no significant differences were observed (p > 0.05) among treatments.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In ABM, LpA1, LpA2, and LpA3 decreased the pH from 6.26 to 5.80-5.95 after 24 h of fermentation, and there was no further pH reduction after 48 h of incubation.

    “In ABM, LpA1, LpA2, and LpA3 decreased the pH from 6.26 to 5.80–5.95 after 24 h of fermentation, and there was no further pH reduction after 48 h of incubation.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In maize silage, LpAv, LpS13, and LpM15 produced significantly lower ammonia-N content compared to control, with no differences among inoculated silages.

    “LpAv, LpS13, and LpM15 produced significantly lower ammonia-N content (4.56 ± 0.05, 4.51 ± 0.30, 4.91 ± 0.10%, respectively) compared to control (5.83 ± 0.11%), with no differences among inoculated silages (Table 7).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    For LpS13 and LpM15 in maize silage, no pH differences were observed between doses 104 and 105 CFU/g (p = 0.581 and 0.842, respectively).

    “however, no differences were observed between the doses 104 and 105 CFU/g (p = 0.581 and 0.842, for LpS13 and LpM15, respectively).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    At silo manufacture, DM ranged from 42.6 ± 0.4% to 43.6 ± 1.5%, without significant differences among samples (p > 0.05).

    “When silos were manufactured, DM ranged from 42.6 ± 0.4% to 43.6 ± 1.5%, without significant differences (p > 0.05) among samples.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The homo- and hetero-fermentative species with the highest frequency of isolation in this study were L. pentosus/plantarum and L. brevis, respectively.

    “The homo- and hetero-fermentative species with the highest frequency of isolation were L. pentosus/plantarum and L. brevis, respectively.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    LpS13 and LpM15 were both able to control yeasts and molds in oat silage.

    “LpS13, derived from sorghum, and LpM15, derived from maize, were effective strains for the acidification of oat silage and were both also able to control yeasts and molds.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Similar to oat silage, improvements in the nutritional value and pH decrease of the forage can be observed despite the fact that epiphytic LAB content in fresh maize was higher than the inoculation rate.

    “Similar to oat silage, these improvements in the nutritional value and pH decrease of the forage can be observed despite the fact that epiphytic LAB content in fresh maize was higher than the inoculation rate.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    LpAv accelerated the pH drop and decreased the proteolysis process in lucerne.

    “LpM15 derived from maize was the most effective strain for oat ensiling and would be the strain of choice for sorghum silage, whereas LpAv, isolated from oat, showed a better performance than the other strains in lucerne, both accelerating the pH drop and decreasing the proteolysis process.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Lactiplantibacillus plantarum subsp. plantarum, Pediococcus pentosaceus and Enterococcus faecium are probably the homofermentative LAB species most intensively used as silage inoculants.

    “Lactiplantibacillus plantarum subsp. plantarum (formerly Lactobacillus plantarum, Zheng et al., 2020), Pediococcus pentosaceus and Enterococcus faecium are probably the homofermentative lactic acid bacteria (LAB) species most intensively used as silage inoculants (Ogunade et al., 2019).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Ogunade et al., 2019 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Heterofermentative LAB, mainly Lentilactobacillus buchneri subsp. buchneri, have been used for more than two decades so far for silage preservation.

    “In order to overcome this problem, heterofermentative LAB, mainly Lentilactobacillus buchneri subsp. buchneri, have been used for more than two decades so far for silage preservation (Filya, 2003; Schmidt et al., 2009).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Filya, 2003 source

      Not checked in the free run.

    • cites Schmidt et al., 2009 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    This is the study where the most diverse variety of forages was used within the same work (15 different plant materials), to isolate new LAB strains for preliminary characterization and future exploitation.

    “To the best of our knowledge, this is the study where the most diverse variety of forages was used within the same work (15 different plant materials), to isolate new LAB strains for preliminary characterization and future exploitation.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    L. plantarum LpM15 and Limosilactobacillus fermentum LfM1 showed potential to be used as inoculants.

    “Based on the studies made, L. plantarum LpM15 and Limosilactobacillus fermentum LfM1 showed potential to be used as inoculants”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The importance of local research and evaluation of new strains adapted to the fermentative process of local forages was recently highlighted.

    “Then, the importance of local research and evaluation of new strains adapted to the fermentative process of local forages was recently highlighted (Amaral et al., 2020).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Amaral et al., 2020 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    When silos are opened and the ensiled material is exposed to the air, a deterioration process may begin, mainly by lactate assimilating yeasts, which leads to an increase in the silage temperature followed by an increase in the pH values, allowing the growth of other aerobic microorganisms and further deterioration.

    “When silos are opened and the ensiled material is exposed to the air, a deterioration process may begin, mainly by lactate assimilating yeasts, which leads to an increase in the silage temperature followed by an increase in the pH values, allowing the growth of other aerobic microorganisms and further deterioration (Kung et al., 2018).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Kung et al., 2018 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    In Argentina, the ensiling market is highly dominated by products manufactured abroad with foreign strains, being the European countries the ones leading the market with a share of 44% of the global silage additives market.

    “In Argentina, the ensiling market is highly dominated by products manufactured abroad with foreign strains, being the European countries the ones leading the market with a share of 44% of the global silage additives market (Fabiszewska et al., 2019).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Fabiszewska et al., 2019 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Interpretationown data

    Although statistical differences were observed among pH values, these were very small and may be not relevant for practical purposes.

    “Although statistical differences were observed among pH values, these were very small and may be not relevant for practical purposes.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Epiphytic LAB widely vary in composition and numbers in plant materials (101-107 CFU/g), depending on many environmental factors.

    “Epiphytic LAB widely vary in composition and numbers in plant materials (101–107 CFU/g), depending on many environmental factors (Pahlow et al., 2003).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Pahlow et al., 2003 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    The strains quickly reached the stationary phase, impairing the proliferation of the epiphytic microbiota, and entered the dead phase.

    “For example, in samples inoculated with 107 CFU/g, total counts after 30 days of fermentation were fairly low, suggesting that the strains quickly reached the stationary phase, impairing the proliferation of the epiphytic microbiota, and entered the dead phase.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    When favorable conditions of anaerobiosis, water activity, and temperature occur, spontaneous lactic acid fermentation may take place on plant material, with LAB becoming dominant in the fermented substrate.

    “However, when favorable conditions of anaerobiosis, water activity, and temperature occur, spontaneous lactic acid fermentation may take place on the plant material, becoming LAB the dominants of the fermented substrate (Di Cagno et al., 2013).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Di Cagno et al., 2013 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    The different growth kinetics of isolates from the same plant material indicate intrinsic diverse growth capabilities that justify strain screening.

    “indicating intrinsic diverse growth capabilities that justify strain screening.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    This adaptation is crucial for survival because it stimulates the production of additional energy and lowers the stress level, e.g., through alkalization of the cytosol under acidic conditions.

    “This adaptation is crucial for survival because it stimulates the production of additional energy and lowers the stress level, e.g., through alkalization of the cytosol under acidic conditions (Papadimitriou et al., 2016).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Papadimitriou et al., 2016 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    In previous reports, a mean DM after ensiling slightly lower than the initial mean value was observed.

    “In previous reports, a mean DM after ensiling, slightly lower than the initial mean value, was observed (Whiter and Kung, 2001; Rizk et al., 2005; Liu et al., 2016).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Whiter and Kung, 2001

      Not checked in the free run.

    • cites Rizk et al., 2005 source

      Not checked in the free run.

    • cites Liu et al., 2016 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Less duplication cycles are needed to reach the stationary phase when inoculating at 107 CFU/g, and in consequence less time is available to develop resistance due to exposure to environmental stresses.

    “The latter concentration is closer to the one commonly reached by LAB at the stationary phase (ca. 108 CFU/g), then less duplication cycles are needed to reach the stationary phase, and in consequence less time is available to develop resistance due to exposure to environmental stresses.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The concentration of 107 CFU/g is closer to the one commonly reached by LAB at the stationary phase (ca. 108 CFU/g).

  • Classified as probably not a substantial claim.

    Premisecitation

    Ammonia-N is generally used as an indicator of silage protein degradation.

    “This parameter is generally used as an indicator of silage protein degradation (Whiter and Kung, 2001).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Whiter and Kung, 2001

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    L. plantarum is the most prevalent species in commercial inoculants.

    “In fact, L. plantarum is the most prevalent species in commercial inoculants (Fabiszewska et al., 2019).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Fabiszewska et al., 2019 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Lactiplantibacillus plantarum is extensively used as an inoculant for ensiling of lucerne, sorghum, maize, ryegrass, and other forages.

    “Lactiplantibacillus plantarum is a species extensively used as inoculant for ensiling of lucerne (Filya et al., 2007; Zhao et al., 2020), sorghum (Thomas et al., 2013), maize (Dogi et al., 2013; Herrmann et al., 2015), ryegrass (Srigopalram et al., 2017; Muthusamy et al., 2020), and other forages (Ávila et al., 2010; Ni et al., 2015).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Filya et al., 2007 source

      Not checked in the free run.

    • cites Zhao et al., 2020 source

      Not checked in the free run.

    • cites Thomas et al., 2013 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Maize is the main forage material used for ensiling in Argentina and worldwide.

    “maize, the main forage material use for ensiling in Argentina and worldwide (Weinberg and Ashbell, 2003; Khan et al., 2015)”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Weinberg and Ashbell, 2003

      Not checked in the free run.

    • cites Khan et al., 2015 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    pH values reported after fermentation with L. plantarum ranged from 4.29 to 5.01.

    “However, L. plantarum has been successfully used over a wide range of DM, pH values reported after fermentation ranged from 4.29 to 5.01 (Filya et al., 2007; Ogunade et al., 2016).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Filya et al., 2007 source

      Not checked in the free run.

    • cites Ogunade et al., 2016 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The strategy of inducing lactic fermentation to promote LAB growth has been largely used for isolating novel LAB candidates for forage fermentations such as ensiling.

    “The strategy of inducing lactic fermentation to promote the growth of LAB has been largely used for the isolation of novel LAB candidates for their exploitation in forage fermentations, such as ensiling (Burns et al., 2018)”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Burns et al., 2018 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    L. plantarum has been successfully used over a wide range of DM.

    “However, L. plantarum has been successfully used over a wide range of DM, pH values reported after fermentation ranged from 4.29 to 5.01 (Filya et al., 2007; Ogunade et al., 2016).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Filya et al., 2007 source

      Not checked in the free run.

    • cites Ogunade et al., 2016 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationnone given

    The lower DM induced by LpM15 use might be due to a slow pH drop.

    “The lower DM induced by LpM15 use, which might be due to a slow pH drop, is an unfortunate event as it could lead to a DM lost (Thomas et al., 2013).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Ensiling lucerne may be challenging because of the low content of water-soluble carbohydrates and its high buffering capacity.

    “Ensiling lucerne may be challenging because of the low content of water-soluble carbohydrates and its high buffering capacity (Pang et al., 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Pang et al., 2011 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation

    These results agreed with the previous dose-response assay in maize, in line with the in vitro to in situ correlation observed by Saarisalo et al. (2007).

    “These results agreed with the previous dose-response assay in maize, in line with the in vitro to in situ correlation observed by Saarisalo et al. (2007).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Saarisalo et al. (2007) source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisereasoning

    When a difference is not statistically significant, one cannot conclude that a negative loss of DM was indeed an increase of DM during ensiling; there was just preservation of DM.

    “When a difference is not statistically significant, then we cannot conclude that a negative loss of DM was indeed an increase of DM during ensiling, there was just preservation of DM.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Lentilactobacillus buchneri is presently the gold standard to promote aerobic stability in corn silage or Napier grass.

    “Lentilactobacillus buchneri is presently the gold standard to promote aerobic stability in corn silage (Santos et al., 2013; Da Silva et al., 2014) or Napier grass (Guan et al., 2020).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Santos et al., 2013 source

      Not checked in the free run.

    • cites Da Silva et al., 2014 source

      Not checked in the free run.

    • cites Guan et al., 2020 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Acetic acid is one of the main organic acids produced by heterofermentative LAB and it has the capacity of promoting aerobic stability when silos are opened.

    “Acetic acid is one of the main organic acids produced by heterofermentative LAB and it has the capacity of promoting aerobic stability when silos are opened (Danner et al., 2003).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Danner et al., 2003 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Although L. buchneri is the main species used in commercial inoculants to improve aerobic stability, some strains may produce biogenic amines, which is an undesirable characteristic.

    “Although L. buchneri is the main species used in commercial inoculants to improve aerobic stability, some strains may produce biogenic amines, which is an undesirable characteristic (Nishino et al., 2007) that justify the research of other heterofermentative LAB (Muck et al., 2018).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Nishino et al., 2007 source

      Not checked in the free run.

    • cites Muck et al., 2018 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisereasoning

    A mean DM after ensiling slightly lower than the initial mean value may occur when initial and final individual values are close among them.

    “This may occur when initial and final individual values are close among them. Then, the differences of the means may be negative, but not statistically significant, and so the loss of DM may look negative.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Wild strains of Lactobacillus hilgardii were proposed for this aim too.

    “Wild strains of Lactobacillus hilgardii were proposed for this aim too (Reis et al., 2017).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Reis et al., 2017 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    In a recent work, two L. plantarum strains were employed for sorghum silages (a commercial one and another isolated from sorghum) and both strains preserved the silages overall quality.

    “In a recent work (Alhaag et al., 2019), two L. plantarum strains were employed for sorghum silages (a commercial one and another isolated from sorghum) and, although both strains preserved the silages overall quality, the commercial strain was not found viable after 30 days of ensiling.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Alhaag et al., 2019 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    These pH values are commonly achieved in the first hours of fermentation, even without the use of microbial inoculants, so there may not be too much room for accelerating the process.

    “These pH values are commonly achieved in the first hours of fermentation, even without the use of microbial inoculants, so there may not be too much room for accelerating the process (Nkosi et al., 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Nkosi et al., 2011 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation

    Our results support the fact that the origin of the strain is of secondary importance, being its own ensiling capacity and the silage material characteristics what matters.

    “Our results support the fact that the origin of the strain is of secondary importance, being its own ensiling capacity and the silage material characteristics what matters (Ávila et al., 2011).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Ávila et al., 2011

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    There are some reports which show that L. brevis enhanced the acetic acid content and aerobic stability of maize silages compared to control but less efficiently than L. buchneri.

    “There are some reports which show that L. brevis enhanced the acetic acid content and aerobic stability of maize silages compared to control but less efficiently than L. buchneri (Danner et al., 2003; Acosta Aragón et al., 2012).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Danner et al., 2003 source

      Not checked in the free run.

    • cites Acosta Aragón et al., 2012 source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Interpretationreasoning

    LpM15 would be the strain of choice for sorghum silage.

    “LpM15 derived from maize was the most effective strain for oat ensiling and would be the strain of choice for sorghum silage, whereas LpAv, isolated from oat, showed a better performance than the other strains in lucerne, both accelerating the pH drop and decreasing the proteolysis process.”
    Quote in the paperSubstantial claim
60 of 60

  • The paper relies on this without support.

    The growth stage at harvesting for each forage was proper for ensiling.

    “Samples (1 Kg) were harvested at an adequate physiological state for silage, indicated by the local farmers”
    “Growth stage at harvesting, for each forage, was the proper one for ensiling according to the farmer that provided the material.”

  • The paper relies on this without support.

    In the authors' region, lucerne, oat, sorghum, and maize are the crops most frequently used for ensiling.

    “In our region, lucerne, oat, sorghum, and maize are the crops more frequently used for ensiling and then attention was focused on these crops.”

  • The paper relies on this without support.

    The concentration of 107 CFU/g is closer to the one commonly reached by LAB at the stationary phase (ca. 108 CFU/g).

    “The latter concentration is closer to the one commonly reached by LAB at the stationary phase (ca. 108 CFU/g), then less duplication cycles are needed to reach the stationary phase, and in consequence less time is available to develop resistance due to exposure to environmental stresses.”

  • The paper relies on this without support.

    A fast decrease of pH close to 4.2 or less is adequate for maize preservation.

    “It is generally considered that a fast decrease of pH close to 4.2 or less is adequate for maize preservation.”

  • The paper relies on this without support.

    Once silos are opened, fermented material is exposed to air and yeasts fermentation may occur.

    “Once silos are opened, fermented material is exposed to air and yeasts fermentation may occur.”

  • The paper relies on this without support.

    L. fermentum has never been employed in commercial inoculants as aerobic stability promoter in the southern cone.

    “To the best of our knowledge, L. fermentum has never been employed in commercial inoculants as aerobic stability promoter in the southern cone.”
6 of 6

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Additives, particularly inoculants containing lactic acid bacteria like Lactobacillus buchneri, are crucial for enhancing fermentation, controlling undesirable microbial activity, and improving overall preservation and nutritive value of silage.

    “Additives, particularly inoculants containing lactic acid bacteria like Lactobacillus buchneri, are crucial for enhancing fermentation, controlling undesirable microbial activity, and improving the overall preservation and nutritive value of the silage [4,9].”

    From Influence of Ensiling Timing and Inoculation on Whole Plant Maize Silage Fermentation and Aerobic Stability (Preliminary Research) (2024) · cites this paper as [9]

    The Introduction and Discussion describe L. buchneri inoculants as used for silage preservation, and the Discussion reports that selected LAB strains improved fermentation, controlled yeasts and molds, or improved indicators of nutritional quality. But the paper does not establish that additives are crucial: its Discussion says maize can reach a preservative pH even without microbial inoculants, and the reported benefits vary by strain. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

  • The absence of statistical significance in LAB numbers among treatments tested was probably due to the typically high numbers of epiphytic LAB (<10^5 CFU g^-1) in the fresh crop.

    “The absence of statistical significance in LAB numbers among treatments tested was probably due to the typically high numbers of epiphytic LAB (<10 5 colony-forming units g -1 ) in the fresh crop [13,54].”

    From The Effects of Short-Time Delayed Sealing on Fermentation, Aerobic Stability and Chemical Composition on Maize Silages (2023) · cites this paper as [54]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Conflicting endpoint for LfSY1

    Major

    The Results say LfSY1 lost aerobic stability at 224 hours, but the Discussion places it among the remaining strains that maintained stability until the assay ended at 9.75 days (234 hours).

  • Conflicting aerobic-stability result for strain 463

    Major

    The Results group strain 463 among those stable until the experiment stopped, while the Discussion says its silage lost stability after 9.3 days.

  • The aerobic-stability assay lasted 9.75 days, not almost 12

    Major

    The reported 234-hour observation period is 9.75 days. The claim that some silages remained stable for at least 12 days when the experiment stopped therefore exceeds the experiment's duration.

    Reported 12 · recomputed 9.75

  • Lucerne-derived strain is labeled LpA03 and LpA3

    Minor

    The Methods name the lucerne-derived strain LpA03, but the Results and tables name it LpA3. The paper does not distinguish these as separate strains.

  • Table 6 promises day-2 results but contains no day-2 row

    Minor

    The table title says it presents analyses after 2 and 30 days, while its time rows show only the beginning and day 30.

5 of 5

Cites

Flagged first, then by citations

1 of the 60 references couldn’t be matched to a known paper.

Research works printed but not linked: 1

  1. Ávila, 2011, Potential use of native microorganisms strains of forage for silage production, Proceedings of the II International Symposium on forage quality and conservation, 25

Cited by

Sorted by citations

Related

Authors
Puntillo, M., Gaggiotti, M., Oteiza, J. M., Binetti, A., Massera, A., & Vinderola, G.
Affiliations
  • Consejo Nacional de Investigaciones Científicas y Técnicas (AR)
  • National Agricultural Technology Institute (AR)
Corresponding author
Vinderola, Gabriel
ORCID
Venue
Frontiers in Microbiology
Publisher
Frontiers Media SA
Published
2020-12-08
Volume
11
Pages
586716-586716
ISSN
1664-302X (ISSN-L)
DOI
10.3389/fmicb.2020.586716
PMC
PMC7752947
OpenAlex
W3112596707
Cited by
65
References
60
Citations by year
2026: 6 · 2025: 17 · 2024: 11 · 2023: 12 · 2022: 11 · 2021: 8
FWCI
9.69
Type
journal article
Language
en
Open access
Licence
Locations
  • Frontiers in Microbiology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • LA Referencia (Red Federada de Repositorios Institucionales de Publicaciones Científicas) (acceptedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • LA Referencia (Red Federada de Repositorios Institucionales de Publicaciones Científicas) (submittedVersion, cc-by-nc-sa)
  • Greater South Information System
Funders
Consejo Nacional de Investigaciones Científicas y Técnicas
Topics
  • Life Sciences › Agricultural and Biological Sciences › Agronomy and Crop Science › Ruminant Nutrition and Digestive Physiology
  • Life Sciences › Agricultural and Biological Sciences › Food Science › Probiotics and Fermented Foods
  • Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Nutrition and Physiology
Keywords
lactic acid bacteria, silage inoculants, aerobic stability, corn silage, Lactobacillus plantarum, Lactobacillus fermentum, heterofermentative bacteria, acetic acid production, forage, silage fermentation quality, autochthonous strains

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-09.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-09.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review