Title

Microbiome and nutrient quality of sugarcane mill/press mud as bio-fertilizer and bio-stimulant

Author

Sophie Minori Uchimiya

Affiliation

U.S. Department of Agriculture (USDA), Agricultural Research Service (ARS), Southern

Regional Research Center, 1100 Robert E. Lee Blvd., New Orleans, Louisiana 70124

Abstract

Bio-fertilizer and bio-stimulant could be developed from sugarcane mill mud/filter cake to
provide (1) carbon source for natural micro-organisms to promote soil health and (2) controlled-
release of nitrogen, potassium, phosphorus, and other essential plant nutrients to reduce the cost
to purchase imported inorganic fertilizers. In Louisiana, waste management for muds became
expensive at mills, as on-site waste piling space is becoming exhausted. Research is needed to
identify the quality of organic carbon and microbiome in mill mud. This study used
bioinformatics to statistically evaluate complex microbial community in mill mud, and to
understand their environmental fate. Fresh mill mud was enriched with heat- and drought-
tolerant microbiota, which became depleted after prolonged environmental exposure (soil
amendment, composting). The rate of environmental transformation is the key to fertilizer value,
as chemical and microbial aging occurs concurrently. In addition, biochemical, thermal, and
electrochemical augmentation approaches can be used to add values to biostimulants from mill
mud. Organic waste-to-biofertilizer concept of mill muds offers a real world message, beyond
United Nation’s Sustainable Development Goals, to diversifying, sustainability-conscious
consumers. Those approaches also address U.S. Department of Agriculture’s Agricultural
Innovation Agenda to “increase the productivity of U.S. agriculture by 40%, while cutting the
environmental footprint of U.S. agriculture in half by 2050”.

Introduction

Mill mud is the solid byproduct from producing raw sugar from harvested sugarcane plants.
Sugarcane mill mud/filter cake is enriched with (1) organic carbon and natural microorganisms
to benefit soil health and (2) phosphorus, potassium, nitrogen (PKN) and other essential plant
nutrients [1] to reduce the cost to purchase inorganic fertilizer. Mill mud is applied to the
sugarcane fields in Florida and Puerto Rico. In Florida, mill mud is used to replicate the fertility
level of organic “muck soil” (Histosols), which is becoming depleted, because of the microbial
oxidation of soil organic carbon [2]. However, there are much wider potential applications for
sugarcane mill mud in the U.S. agriculture. In March of 2022, USDA announced $250 million
investment from commodity credit corporation funds to “support innovative American-made
fertilizer to give US farmers more choices in marketplace”. This initiative originated from the
price hike (more than double relative to last year), foreign dependence (U.S. is among the top

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PKN importer globally), and lack of competition in fertilizer market. Biofertilizer and
biostimulants could be developed, tested, and deployed from mill mud to enhance soil health and
nutrient use efficiency within the framework of best management practice adoption.

Biostimulants are used to enhance crop yields by means other than the PKN fertilizer values, e.g.,
by enhancing resistance to disease, pest, and stress; increasing bioavailability of nutrients in soil;
and by releasing molecules that act as plant hormones directly or indirectly. Mill mud could be
engineered via biochemical, thermal, and electrochemical conversion methods to achieve the
target specs of biostimulants. Currently available biostimulants are limited primarily to humic
substances, microbial inoculants, or seaweed and protein extracts [3]. Humic substances and
soil’s native microbes (and their exudates) are examples of natural soil components capable of
stimulating plant growth [4].

Soil application of sugarcane mill mud will benefit Louisiana agriculture, beyond sugarcane, in
the near term by offering free, renewable, locally-sourced organic fertilizer to offset the capital
cost. For example, soybean makes up the largest export market of $700 million in Louisiana
(USDA-ERS, 2018); yet, $27 million per year is spent to import PKN chemical fertilizer. Global
supply of phosphorus and other imported, fossil-based chemical fertilizer is predicted to become
depleted in the coming decades. Longer-term benefits to soybean and other commodity
production will include the following: (1) diversified foreign and domestic marketing strategy to
target health- and sustainability-conscious consumers; (2) soil health promotion by mill mud’s
ability to supply beneficial microbial community and controlled-release of nutrients; and (3)
locally-sourced farming practice to mutually benefit soybean and sugarcane growers by reducing
capital costs on fertilizers and waste management.

Materials and Methods

A total of 15 mill mud, bagasse, and soil samples were analyzed chemically and by 16S rRNA
gene sequencing [5]. Greenhouse experiments were conducted at 10, 20, 30, and 40 g mill mud
per kg soil application rates with 10 replicates in a randomized block design to test germination
rate, yield, and biomass weight for soybeans.

Results and Discussion

Figure 1 shows representative mill mud, bagasse, and soil samples to illustrate heterogeneous
nature of field samples. Statistical analysis of various chemical parameters (% solid, % organic
matter, pH, soluble salts, C/N, ammonium, nitrate, P, K, Ca, Mg, Na, Fe, Cu, Mn, Na, Al, and
ash contents) afforded chemical signatures to distinguished mill mud (high P 2O5) from bagasse
(high C/N) and soil (high pH) samples of diverse geographical/environmental origins (Figure 2).

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Figure 1. Illustrative examples of mill mud, bagasse, and soil samples for chemical and
microbial characterization.

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Figure 2. Chemical classification of soil, bagasse, and mill mud samples.

Chemically distinguished sample groups (mill mud, bagasse, and soil in Figure 2) were then
analyzed for microbial composition in Figure 3. Factory-generated (under elevated
temperatures) mill mud was uniquely enriched with heat/drought/cold tolerant microbes (Figure
3). Upon environmental exposure, microbial composition of mill mud diversified to include
community often found in soils. Similar changes in microbial composition were observed for
bagasse as a result of aging in open fields. The results suggest rapid diversification of factory-
generated sugarcane processing byproducts as a result of aging, which include natural
composting in a waste pile, outdoor storage until next campaign season, and soil amendment.

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Figure 3. Microbial composition of mill mud, bagasse, and soil samples.

Figure 4 summaries how chemical and microbial composition of mill mud could be used to trace
the environmental fate of mill mud after application to the field. First, samples are chemically
categorized for high mineral (soil), high organic (bagasse), and high nutrient (mill mud) contents
in Figure 4, left. Samples that contain mill mud may be chemically categorized as soil, because
of high mineral contents, e.g., soil amended with mill mud or legacy waste pile with high sand
content. Next, each class (mill mud, bagasse, or soil) is examined for microbial composition to
understand their environmental fate. Upon environmental exposure, factory-generated microbial
community diversified to resemble the microbial composition of soil samples.

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Figure 4. Methods to trace environmental fate of sugarcane mill mud after soil application.

Preliminary greenhouse experiments on soybeans indicated no statistically significant effects of

mill mud on germination rate, under the standard fertilization rate (Figure 5). Much like other
sustainable farming practices (e.g., cover crops, allelochemicals, mulching, green manure),
growers are more likely to incorporate mill mud application, if it does not negatively impact crop
yield and quality [6].

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Figure 5. Gemination rate of soybean grown in pots (10 replicates) at 0-40 g mill mud per kg
soil application rates.

References

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