HyTide

The science behind D68

A surfactin-family lipopeptide, produced by a selected Bacillus strain and screened for one job: keeping pathogenic bacteria from taking hold in the gut.

How it works

D68's core molecule is a cyclic lipopeptide: a ring of seven amino acids linked to a beta-hydroxy fatty acid. It acts on bacteria in two ways, depending on concentration.

At low levels it interferes with quorum sensing, the chemical signals bacteria use to coordinate and build biofilms, so colonies struggle to form. At higher levels it penetrates mature biofilms, breaks bacterial membranes and disperses what remains, which stops pathogens from attaching to the gut wall.

Conventional antimicrobial peptides punch holes in cell membranes through electrical charge. Used long-term, that single mode of action can select for resistant strains. D68's biofilm and signalling route avoids that trap.

Diagram showing D68 inhibiting biofilm formation, penetrating mature biofilm and dispersing it
Concentration-dependent action: inhibit early biofilm, then penetrate, rupture and disperse mature biofilm.

How the strain was found

D68 came out of a systematic screening program that combined multi-omics with layered functional testing, so strain selection was driven by data rather than chance.

  1. Sample & sequence

    High-throughput sequencing to profile microbial diversity and spot differential microbiota.

  2. Isolate

    Target strains isolated and grown in pure culture under controlled conditions.

  3. Test function

    Phenotypic and functional assays for the traits that matter in production.

  4. Identify

    Molecular sequencing confirms strain identity and genetic uniqueness.

  5. Characterize actives

    Key bioactive proteins purified and identified by mass spectrometry.

Antibacterial activity

Measured by agar well diffusion (Oxford cup): D68 is placed in a well and the clear zone where bacteria fail to grow is recorded. Higher dilutions that still produce a zone mean stronger activity.

PathogenRelevanceResult
Escherichia coli K88Swine, poultryInhibited at 1:500 (1:2000 in a second lab)
Salmonella spp.Poultry, food safetyInhibited at 1:500
Staphylococcus aureusPoultry, dairyInhibited at 1:2000
Clostridium perfringensNecrotic enteritisInhibited at 1:2000
Riemerella anatipestiferDucksZone >10 mm at 1:2000; a commercial antimicrobial peptide showed none
Vibrio harveyi, V. parahaemolyticusShrimp, fishZone >18 mm

Tests run independently at several partner laboratories, including a pharmaceutical lab and an international feed company.

Agar plates showing clear inhibition zones against E. coli K88, Salmonella, Staphylococcus aureus and Clostridium perfringens
Left to right: E. coli K88, Salmonella, S. aureus, C. perfringens.
Agar plate showing a clear inhibition zone against Vibrio
Inhibition zone against Vibrio.

Safety

The D68 strain carries no acquired antibiotic resistance. It is fully susceptible to all eight antibiotics tested, with inhibition zones of 23 to 46.5 mm: cefradine, erythromycin, chloramphenicol, clindamycin, norfloxacin, amikacin, carbenicillin and gentamicin.

Animal safety studies found no lesions in the heart, liver, spleen, kidneys, jejunum, ileum or colon. The strain is not genetically modified or gene-edited.

Manufacturing

D68 is produced by submerged liquid fermentation, then concentrated and spray-dried or carrier-dried at low temperature to preserve the active compounds. The finished powder contains at least 1×109 CFU/g of the producing Bacillus together with its metabolites, including lipopeptides, protease and amylase.

The strain grows quickly and tolerates heat and bile salts at 0.1 to 0.3%.

D68 compared with antimicrobial peptides

D68 postbioticTypical antimicrobial peptide
Mode of actionQuorum-sensing interference, biofilm disruption and immune supportCharge-based membrane damage; can select for resistance
SpectrumBroad: Gram-positive and Gram-negativeOften narrow, aimed at specific bacteria
StabilityHeat- and acid-tolerantReadily broken down by proteases
Added valueDigestive enzymes and metabolites support feed efficiencyAntibacterial only
Cost in useLow production cost, competitive inclusion costExpensive to produce

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