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Wet Pile Microbiome: The 45-Day Fermentation Timeline

Direct Answer: During 45-day Shou Puerh fermentation (Wo Dui), microbial succession follows: Days 1-5: Environmental colonizationDays 5-15: Yeast/Bacteria bloomDays 15-30: Mold dominance & peak heatDays 30-45: Stabilization & cooling. Each phase produces different compounds that collectively transform green leaves to dark, smooth tea.

This timeline is not linear—temperature, humidity, and pile turning affect microbial succession. Understanding the phases helps identify when fermentation is on-track or failing.

Cross-section of Wo Dui fermentation pile showing microbial succession over 45-day Puerh Shou fermentation timeline

The microbial Succession: An Ecological Timeline

A fermentation pile isn't random chaos. It follows predictable ecological succession—like how bare rock becomes lichen, then moss, then plants over decades. In tea, this happens over 45 days.

This succession is driven by:

Phase 1: Inoculation (Days 1-5) - The Environmental Stage

Temperature: 20-25°C | pH: 5.5-6.0 | Dominant Organisms: Environmental microbes

What's Happening

Freshly processed green tea leaves are moisture-adjusted (water added to ~70% humidity) and piled in a fermentation chamber. The pile is inoculated naturally—using microbes from the air, the processing equipment, previous fermentations, or added culture.

These first organisms are primarily:

Microbial Activity Level: Low

Little fermentation is occurring. Microbes are simply establishing on the substrate. Temperature remains ambient because microbial respiration hasn't yet accelerated. The pile smells fresh, like moist green tea.

Phase 2: Enzyme Release & Fermentation (Days 5-15) - The Yeast-Bacteria Stage

Temperature: 25-40°C (rising) | pH: 4.8-5.5 (declining) | Dominant Organisms: Yeasts and lactic acid bacteria

What's Happening

Environmental microbes begin exponential growth. Aspergillus niger starts enzyme production, degrading cell walls. Yeasts ferment simple sugars anaerobically, producing ethanol and CO₂. Lactic acid bacteria (LAB) produce organic acids, lowering pH.

Temperature Rising

Microbial metabolism generates heat. If environmental controls are good (humidity ~70%, pile turning), temperature rises steadily. By Day 10-12, the pile's interior reaches 30-35°C. This heat acceleration is critical—it selects for thermophilic organisms and accelerates chemical reactions.

Aroma Change

Days 5-8: Slightly fruity, yeasty aroma emerges (ethanol production). Days 10-15: Increasingly "fermented" smell—sour, slightly vinegary notes as LAB organic acids accumulate.

Microbial Activity Level: Rising

This is an active, rapid growth phase. Doubling times for bacteria are 20-30 minutes. The community is still diverse, but yeasts and LAB are dominant.

Phase 3: Thermophilic Peak (Days 15-30) - The mold-Dominated Heat Stage

Temperature: 45-60°C (peak, Days 18-25) | pH: 3.5-4.5 (acidic) | Dominant Organisms: Aspergillus niger, Blastobotrys, thermophilic bacteria

What's Happening

This is the critical transformation phase. Aspergillus niger and Blastobotrys adeninivorans reach peak activity. Temperature rises to 50-60°C (often with hotspots exceeding this).

The Heat Generation

Blastobotrys adeninivorans performs aerobic respiration at maximum rate. Each cell uses glucose to generate ATP, releasing massive amounts of heat (exothermic fermentation). This is why experienced fermenters must turn the pile regularly—to prevent thermal runaway exceeding 65°C (which kills desired organisms and creates off-flavors).

Chemical Transformation Accelerates

At these elevated temperatures, catechins oxidize to theaflavins and theabrownins at 10-100x normal rates. Maillard reactions occur (browning), creating dark color and savory compounds. Volatile aromatics condense or evaporate—determining final tea aroma profile.

Color Change: Green to Brown

By Day 20-22, visual inspection shows dramatic darkening. Fresh green leaves become brown. By Day 25-28, they're dark brown to nearly black. This is the visible marker of successful fermentation.

Aroma Peak

Days 15-20: Rich, earthy, mushroom-like aroma. Days 20-30: Increasingly mellow, less sharp. The peak fermentation smell is quite strong—some describe it as "petrichor" (earthy after rain) or "forest floor."

Microbial Activity Level: Peak

Maximum metabolic activity. Enzyme production is highest. If turning is poor or environmental control fails, temperature can exceed safe limits, and pathogenic bacteria may colonize.

Phase 4: Cooling & Stabilization (Days 30-45) - The Bacterial Transition

Temperature: 35-45°C (declining) | pH: 3.5-4.2 (stabilized) | Dominant Organisms: Thermophilic bacteria, residual molds

What's Happening

Temperature naturally declines as the pile's nutrient-rich substrate is progressively depleted. Yeasts and Blastobotrys slow their activity. Bacillus and thermophilic actinomycetes (heat-loving bacteria) become dominant. These organisms consolidate the chemical transformations and stabilize flavor compounds.

Final Aroma Development

Days 30-40: The sharp fermentation aroma mellows. Pleasant fruity, woody notes emerge. Days 40-45: The tea smells "aged"—earthy, slightly sweet, mellow.

Flavor Stabilization

This phase locks in the flavor profile. Late-stage microbial enzymes create the final compound cocktail. Lovastatin production peaks during this phase. Remaining polyphenols stabilize in their oxidized forms.

Microbial Activity Level: Declining

Growth slows, then plateaus. The community stabilizes into a mature state. By Day 40, significant activity has ceased. The pile is considered "fermented" by professional standards at Day 35-40, though some makers extend to Day 45 for additional aging.

Detailed Fermentation Timeline Table

Days Temp (°C) Phase Dominant Microbes Key Changes
1-5 20-25 Inoculation Environmental microbes, spores Colonization. Mild moisture absorption. No color change.
5-10 25-32 Yeast Bloom Yeasts (S. cerevisiae, others), early Aspergillus Temperature rises. Slight fruity aroma. Early color darkening.
10-15 32-42 LAB Dominance Lactic acid bacteria, Aspergillus niger Rapid enzyme production. Sour aroma. Continued darkening. pH drops.
15-20 42-55 Heat Peak Begins Blastobotrys adeninivorans, A. niger (peak) Rapid temperature rise. Earthy aroma. Color turns to dark brown. Major polyphenol oxidation.
20-25 50-60 Heat Peak Blastobotrys (peak), thermophilic bacteria Maximum temperature. Maillard reactions. Deep brown to black color. Rich aroma.
25-30 45-55 Heat Decline Begins Blastobotrys (declining), Bacillus (rising) Temperature moderates. Flavor compounds stabilize. Aroma mellows. Lovastatin production peak.
30-35 40-50 Bacterial Phase Bacillus, thermophilic actinomycetes Continued cooling. Aroma becomes aged and mellow. Chemical reactions slow. Fermentation largely complete.
35-45 35-42 Stabilization Thermophilic bacteria (Bacillus, others) Final consolidation. Flavor stabilizes. Aroma fully developed. Microbial activity plateaus. Ready for rest and packaging.

The Critical Window: Days 20-25

This is where fermentation success or failure is determined. If temperature reaches 65°C+, unwanted microbes colonize and the tea develops harsh, off-flavors. If temperature stays below 45°C, fermentation stalls and the tea remains bitter. Professional fermenters monitor this window obsessively.

Environmental Factors That Alter This Timeline

Humidity

Too dry (<60% RH): Fermentation slows. Microbes struggle to metabolize. Timeline extends to 60+ days.

Optimal (65-75% RH): Microbial growth is optimal. Timeline follows schedule.

Too wet (>80% RH): Pathogenic bacteria (Bacillus, Clostridium) may colonize. Off-flavors develop.

Temperature Control

Cool environment (15°C): Timeline extends to 60+ days. Slow fermentation.

Warm environment (25°C): Fermentation accelerates. Timeline may compress to 35-40 days.

Pile Turning

Regular turning (every 3-5 days) aerates the pile, prevents hotspots, and ensures even fermentation. Lack of turning creates anoxic zones and uneven microbial colonization.

How to Identify Fermentation Progress (Practical Signs)

Days 5-10: Slight fruity smell. Leaves beginning to brown. Temperature rising to 28-32°C.

Days 10-15: Sour, fermented aroma. Leaves noticeably darker. Temperature 35-40°C.

Days 15-20: Rich earthy smell. Leaves dark brown. Temperature 45-55°C.

Days 20-25: Peak aroma. Leaves nearly black. Temperature 50-60°C. This is the critical decision point.

Days 25-35: Aroma mellowing. Leaves fully dark. Temperature declining to 40-45°C.

Days 35-45: Aged, mellow aroma. Leaves black. Temperature 35-40°C. Fermentation visually complete.


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