Research · Evidence · Patent
SSF Seaweed Science:
In-Vitro Evidence & Strain Registry
All quantitative claims on this page are preliminary in-vitro results. In-vivo validation is scheduled for Q4 2026. Data is shared to enable scientific review and partnership due diligence — not as final proof of commercial efficacy.
⚗️ In-Vitro: Preliminary
🐄 In-Vivo: Q4 2026 Planned
🔬 Proprietary SSF Process
In-Vitro Methane Inhibition
~50% CH₄ Inhibition:
Gas Production Assay Results
All values are in-vitro only. PRELIMINARY. Not validated in live animals.
~50%
CH₄ inhibition vs. control
Preliminary in-vitro result — batch rumen gas production assay. In-vivo results not yet available.
Preliminary in-vitro result — batch rumen gas production assay. In-vivo results not yet available.
Assay Conditions & Methodology
Gas production assay — batch rumen fermentation in vitro
| Parameter | Value / Description |
|---|---|
| Assay type | Batch rumen fermentation gas production assay |
| Incubation period | 24 hours |
| Temperature | 39°C (rumen physiological temperature) |
| Inoculum | Buffered rumen fluid from rumen-cannulated Holstein cows |
| Substrate | SSF-processed seaweed blend (Eucheuma denticulatum + Kappaphycus alvarezii, proprietary formulation) |
| Dose tested | Proprietary (commercial dosage to be confirmed post in-vivo validation) |
| Control | Basal substrate without seaweed addition |
| Gas measurement | Syringe gas production (total gas); CH₄ measured by gas chromatography |
| Replicates | 3 replicate syringes per treatment; 2 experimental runs |
| Primary outcome | ~50% reduction in CH₄ production (% of total gas vs. control) |
| Secondary outcomes | Total gas volume, volatile fatty acid (VFA) profile, dry matter disappearance (DMD) |
| Conducting institution | Aenon R&D — external verification pending |
| Data status | Internal preliminary results. Peer review not completed. |
Key Preliminary Findings
Reported alongside supporting measurements — not standalone claims
| Measurement | Direction vs. Control | Status |
|---|---|---|
| CH₄ production (% total gas) | ↓ ~50% | Preliminary |
| Total gas volume | Moderate reduction (fermentation continues) | Preliminary |
| Propionate ratio in VFA | ↑ Trend (propionate pathway — H₂ sink) | Monitoring |
| Dry matter disappearance | Maintained (no negative digestibility signal) | Preliminary |
| pH after 24h incubation | Within normal rumen range (6.4–6.8) | Preliminary |
⚠ Propionate A/P ratio trend is YELLOW evidence. Numerical claim not asserted until in-vivo data is available.
Proposed Dual Inhibition Mechanism
🦠
Mechanism 1: Methanogenic Archaea Inhibition
Bioactive polysaccharide fractions from SSF-processed seaweed are proposed to disrupt the cell membrane integrity of methanogenic archaea (primarily Methanobrevibacter ruminantium), directly reducing CH₄-generating metabolic activity. SSF fermentation at 45°C modifies carrageenan structure, potentially enhancing bioavailability of inhibitory compounds.
⚗️
Mechanism 2: H₂ → Propionate Redirection
Fermentation metabolites from L. buchneri OV-5 and W. cibaria KACC 92499P are hypothesised to promote propionate-pathway bacteria, shifting H₂ from methanogenesis (CH₄ production) toward propionigenesis. This represents an alternative H₂ sink that maintains energy yield while reducing methane output.
Note: Proposed mechanisms are based on preliminary in-vitro observations and current literature. Causal confirmation requires in-vivo mechanistic studies (planned Q4 2026).
Research Milestones
Q1 2024
Species Selection
Spinosum + Cottonii selected based on composition screening and gel-formation profile analysis
Q3 2024
Strain Isolation
Fermentation strains isolated and characterised for SSF suitability. Institutional deposit completed.
Q1 2025
SSF Process Optimisation
Key SSF process parameters optimised at bench scale. Process details are proprietary.
Q2 2025
In-Vitro Assay
~50% CH₄ inhibition observed in preliminary batch rumen gas production assay (in-vitro only).
Q3 2025
IP Development
Proprietary SSF process and formulation documentation completed.
Q4 2026
In-Vivo Trial (Planned)
Animal validation study planned to evaluate methane emissions, feed intake, digestibility and animal performance. [Planned]
Microbiology · Strain Registry
Fermentation Strains:
Registration & Functional Profile
Both strains are selected for thermotolerance at 45°C — a critical requirement imposed by the gel-forming property of carrageenan in Eucheuma denticulatum, which prevents liquid-phase fermentation and necessitates solid-state processing.
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Lactobacillus buchneri OV-5
Registration: OV-5 (Internal Designation)
Species: Lactobacillus buchneri (Lb. buchneri) — lactic acid bacterium
Thermotolerance: Active at 45°C — essential for SSF at carrageenan gel-forming temperature
Proposed function: Primary lactic acid fermentation; bioactive compound modification via enzymatic activity; partial acidification of substrate
SSF role: Initiates fermentation cascade; produces acetate and 1,2-propanediol from carbohydrates; modifies substrate matrix
Safety profile: L. buchneri has a long history of safe use in feed fermentation (silage); classified as generally recognised as safe in fermented feed contexts
Status: Internal strain designation OV-5 — KIPO patent application includes strain characterisation data
Thermotolerance: Active at 45°C — essential for SSF at carrageenan gel-forming temperature
Proposed function: Primary lactic acid fermentation; bioactive compound modification via enzymatic activity; partial acidification of substrate
SSF role: Initiates fermentation cascade; produces acetate and 1,2-propanediol from carbohydrates; modifies substrate matrix
Safety profile: L. buchneri has a long history of safe use in feed fermentation (silage); classified as generally recognised as safe in fermented feed contexts
Status: Internal strain designation OV-5 — KIPO patent application includes strain characterisation data
🧫
Secondary Fermentation Strain (Patent-Protected)
KIPO Patent Application Strain 2 · KACC-Registered Patent Strain
Species class: Heterofermentative lactic acid bacterium — thermotolerant at 45°C
Institutional deposit: Formally deposited at the Korean Agricultural Culture Collection (KACC) — National Institute of Agricultural Sciences, RDA
Thermotolerance: Active at 45°C; adapted for solid-state conditions
Proposed function: Secondary fermentation; exopolysaccharide (EPS) production from seaweed polysaccharides; potential prebiotic activity in rumen
SSF role: Synergistic with primary strain; contributes to substrate matrix modification and bioactive compound profile shaping
KACC registration: Deposited with KACC — "P" suffix indicates patent strain deposit; verifiable through KACC database (kacc.rda.go.kr)
Status: Formally registered — institutional deposit confirms existence; strain identity will be publicly disclosed upon patent grant
Institutional deposit: Formally deposited at the Korean Agricultural Culture Collection (KACC) — National Institute of Agricultural Sciences, RDA
Thermotolerance: Active at 45°C; adapted for solid-state conditions
Proposed function: Secondary fermentation; exopolysaccharide (EPS) production from seaweed polysaccharides; potential prebiotic activity in rumen
SSF role: Synergistic with primary strain; contributes to substrate matrix modification and bioactive compound profile shaping
KACC registration: Deposited with KACC — "P" suffix indicates patent strain deposit; verifiable through KACC database (kacc.rda.go.kr)
Status: Formally registered — institutional deposit confirms existence; strain identity will be publicly disclosed upon patent grant
Note on strain identity: Both fermentation strains are formally characterised and documented in the KIPO patent application. Strain identities are patent-protected and will be disclosed upon grant. KACC registration is verifiable through kacc.rda.go.kr. Functional roles are proposed based on species-class literature and preliminary in-vitro observations; in-vivo mechanistic confirmation is pending.
Intellectual Property
KIPO Patent-Pending:
22 Claims Overview
The KIPO (Korean Intellectual Property Office) patent application covers the SSF process, strain combination, and composition. Full patent number will be disclosed upon grant. Claims listed below are structural summaries — exact legal language is in the filed application.
Patent Status
KIPO Application Filed
Korean Intellectual Property Office — Pending Grant
Total Claims
22
Process + Composition + Application claims
Claim Category Summary
1
SSF process at 45°C using Eucheuma denticulatum (Spinosum) as primary substrate
2
Substrate composition: Spinosum 70% + Cottonii 30% blend ratio
3
Strain combination: L. buchneri OV-5 + W. cibaria KACC 92499P co-culture
4
Fermentation duration optimisation (72h primary cycle)
5
Solid-state configuration — gel-formation avoidance vs. liquid fermentation
6–8
Process parameter ranges: temperature, humidity, substrate moisture content
9–12
Composition claims: final product dry matter, bioactive fraction, particle size
13–16
Application claims: dosage range in ruminant feed, target species, inclusion method
17–19
Method claims: enteric methane reduction using the SSF-processed composition
20–22
Dependent claims: Korea–Vietnam supply chain integration, drying specifications
Patent disclosure note: Full application number and independent claim text will be publicly disclosed upon KIPO examination and grant. The 22-claim structure above summarises the scope of protection sought. Patent ownership: Aenon Co., Ltd. is the applicant. External IP co-ownership or licensing arrangements, if any, will be disclosed separately.
Next Stage · In-Vivo
In-Vivo Trial:
Planned Q4 2026
Facility
SNU Pyeongchang Campus
Seoul National University, Pyeongchang
Design
6-Head Crossover
Dairy cows — treatment vs. control periods
Measurement
Respiration Chamber
Direct CH₄ and CO₂ flux measurement
Timeline
Q4 2026 Start
IRB and facility approval process ongoing
In-vivo data is YELLOW evidence — it will be published with full methodology when available. No in-vivo results are available as of the date of this page.
Substrate Biology
Why Spinosum + Cottonii?
Substrate Selection Rationale
The 70:30 Spinosum-to-Cottonii ratio is determined by carrageenan content, gel-forming behaviour, bioactive compound profile, and supply chain availability from Vietnam. This is not arbitrary — the ratio is a core patent claim.
Primary Substrate · 70%
Eucheuma denticulatum
Spinosum — ι-carrageenan dominant
- ι-carrageenan gels at room temperature → liquid fermentation impossible
- Gel formation is why SSF is not merely preferred — it is the only viable process
- High bioactive sulphated polysaccharide fraction
- Commercial scale availability: Vietnam (Ninh Thuận, Khánh Hòa)
- Established supply chain via AENON Vietnam agricultural network
Secondary Substrate · 30%
Kappaphycus alvarezii
Cottonii — κ-carrageenan dominant
- κ-carrageenan — complementary gel-forming profile to ι-carrageenan
- Contributes distinct sulphated polysaccharide fractions to final product
- 30% ratio modulates total substrate viscosity and SSF matrix structure
- Also commercially cultivated in Vietnam — same supply chain
- Both species: bromoform-free (no halogenated compound concerns)
The specific 70:30 ratio and SSF process combination is the subject of patent Claim 2. Variations from this ratio require separate validation.
Scientific Due Diligence & Partnership
Researchers, investors, and institutional partners may request our full internal technical dossier for due diligence review.