1. ProSil M-100
Silage Additive Incorporating a Unique Combination of Lactic Acid Bacteria for Improving Stability of Whole-Crop Cereal, Maize and Crimped Grain Silage after Opening

2. Features & Benefits
Contains 3 unique lactic acid bacteria (LAB) isolated from farm silages: Lactobacillus fermentum, Lactobacillus plantarum and Pediococcus acidilactici.
Improves the stability of whole-crop cereal, maize and crimped grain silages after opening
Increases acetic acid to improve stability after opening
Reduces the number of yeasts which cause stability problems in silage
Improves silage fermentation
Successfully used on farms
Easy to use
Safe
EU notified strains

3. Maize Post-opening problems
Maize is a great forage for silage due to it’s high dry matter and energy contents
The well known problem with maize silage is that it is very susceptible to heating after opening
This heating is mainly caused by yeasts
ProSil M-100 contains Lactobacillus fermentum which produces acetic acid, which inhibits the activities of yeasts

4. Unique Combination Inoculant
There are 2 main types of lactic acid bacteria: homofermentative and heterofermentative
ProSil M-100 contains a combination of: Homofermentative LAB: Lactobacillus plantarum and Pediococcus acidilactici, which improve silage fermentation, and Heterofermentative LAB: Lactobacillus fermentum, which improves the stability of silage

5. Homofermentative Mode of Action
Homofermentative LAB are very efficient at fermenting hexose sugars in forage to lactic acid: 1 glucose or 1 fructose → 2 lactic acid Fermentation of the rarer pentose sugars: 1 arabinose or 1 xylose → 1 lactic acid + 1 acetic acid
This efficient utilisation of sugar results in a fast pH fall, resulting in: - improved silage fermentation - lower losses - improved nutritional value of silage - improved livestock performance

6. Heterofermentative Mode of Action
The mode of action of the heterofermentative Lactobacillus fermentum is similar to that of L. buchneri
Acetic acid is formed from the fermentation of hexose and pentose sugars and from the fermentation of lactic acid
Pathways for fermentation of lactic acid to acetic acid in silage are not fully understood and the pathway shown in slide 7 is one proposed without an external electron acceptor and under anaerobic conditions by Oude Elferink et al Other possible pathways also exist

7. ProSil M-100 Acetic Acid Pathways
From hexose sugars: 3 fructose → 1 lactic acid + 1 acetic acid + 2 mannitol + 1 CO2 1 glucose + 2 fructose → 1 lactic acid + 1 acetic acid + 2 mannitol + 1 CO2
From pentose sugars: 1 arabinose or 1 xylose → 1 lactic acid + 1 acetic acid
From lactic acid: 2* lactic acid → 1 acetic acid + 1 1,2-propanediol + (trace) ethanol + 1 CO2 * Proposed molar levels

8. It Begins with Good Strain Selection (1)
The 2 homofermentative LAB in ProSil M-100 were each isolated from well preserved silage, so they originate from silage and grow in that habitat.
Good selection criteria are essential to ensure that the best lactic acid bacteria are selected for inoculants. These selection criteria included:
Strain safety
Fast growth rate
The ability to ferment silage quickly / fast pH fall
Strong production of lactic acid, with check on when the lactic acid was produced
Non-proteolytic to preserve more protein in silage
No production of extracellular polysaccharides

9. It Begins with Good Strain Selection (2)
The heterofermentative Lactobacillus fermentum was selected from over 600 strains of LAB which were isolated from stable high dry matter silages on dairy farms. It originates from silage and grows in that habitat.
The selection criteria for L. fermentum included:
Strain safety
The ability to ferment lactic acid to acetic acid when presented with lactic acid as the only source of carbon and energy
Non-proteolytic to preserve more protein in silage
No production of extracellular polysaccharides

10. A Combination That Works
Homofermentative LAB are faster growing than heterofermentatives (see slide 11) and would dominate silage fermentation if the wrong ratio of each was used.
Extensive testing on dairy farms has identified the optimum ratio of homofermentatives and heterofermentatives to include in ProSil M-100 to achieve both improved stability and improved silage fermentation and feeding quality

11. Doubling time and Temperature Range
Double in number every: Pediococcus acidilactici 35 minutes Lactobacillus plantarum 33 minutes Lactobacillus fermentum 53 minutes
Temperature range for growth Pediococcus acidilactici °C Lactobacillus plantarum °C Lactobacillus fermentum °C This covers the range of temperatures found in silage

12. Fermentation of the most important (5 and 6 Carbon) Sugars
P. acidilactici
Arabinose
Ribose
Galactose
Glucose
Fructose
Mannose
L. plantarum
Xylose
L . fermentum

13. More Acetic Acid = Improved Aerobic Stability in Maize Silage
Parameter
Control
ProSil M-100
Significance
DM %
32.5
32.2 pH
3.6 ns
Lactic acid % DM
4.2
4.3
Acetic acid % DM
1.3
1.9
P<0.001
Average temperature after opening °C
25.5
24.7
P<0.01
ProSil M-100 significantly increased (P<0.001) acetic acid in maize silage,
leading to a significantly lower (P<0.01) average temperature after opening
Lower temperatures mean that more energy is retained in the silage, which increases feeding value

14. More Acetic Acid = Improved Aerobic Stability in Whole-Crop Wheat Silage
Parameter
Control
ProSil M-100
Significance
DM %
38.6
38.5 pH
4.2
4.1 ns
Lactic acid % DM
3.5
3.8
Acetic acid % DM
1.4
2.3
P<0.001
Average temperature after opening °C
25.6
21.5
ProSil M-100 significantly increased (P<0.001) acetic acid in whole-crop wheat silage, leading to a significantly lower (P<0.001) average temperature after opening

15. ProSil M-100 Reduces Temperature after Opening Trial 3

16. Temperature deg C Change in temperature with time of maize silages10 20 30 1 13 25 37 49 61 73 85 97
109
121
133
145
Temperature deg C
Time hours
Change in temperature with time of maize silages
exposed to air
Untreated
ProSil M-100
Homoferm.
Ambient
Maize dry matter 31.7%

17. ProSil M-100 Reduces Temperature after Opening Trial 4

18. Effect of different treatments on heating of maize silage10 12 14 16 18 20 22 24 1 19 28 37 46 55 64 73 82 91
100
109
118
Time Hours
Temperature deg C
Untreated
ProSil M-100
ambient
Maize dry matter 29.3%

19. Hours to +2°C Rise above Ambient
Trial
Control
ProSil
M-100
Significance 3 21
100
P<0.001 4 30
105
Stability significantly extended (P<0.001) by over 3 days after opening in both trials

20. Reduced Yeast in Silage
Control
ProSil M-100
Yeasts CFU / g
2,100,000
17,000
CFU = colony forming units
Substantial reduction in the number of yeasts at opening
Reduction in yeasts leads to greater stability

21. Specifications
Active ingredients: Lactobacillus fermentum, Lactobacillus plantarum and Pediococcus acidilactici.
Carrier: Highly soluble dextrose
Packed in 100g amounts in alufoil sachets to treat 100 tonnes of whole-crop cereal, maize or crimped grains
All strains have been notified to the European Commission and to the European Food Safety Authority
Storage stability: 18 months from the date of manufacture when stored at ambient temperature (<20°C).

22. Usage instructions
One 100g sachet of ProSil M-100 treats 100 tonnes of whole-crop cereal, maize or crimped grains
Mix the contents thoroughly in a small amount of clean cold tap water
Dilute the solution as necessary in clean cold tap water to suit the applicator and speed of ensiling.
ProSil M-100 can be applied at any application rate between 100 ml and 2 litres per tonne of forage, provided always that 1 g of product is applied per tonne of forage

23. ProSil M-100 Summary
Contains unique lactic acid bacteria isolated from good quality silages and from stable silages
The formulation is based on what works on dairy farms
Lengthens the stability of whole-crop cereal and maize silage after opening
Increases the content of acetic acid
Fewer yeasts recorded in maize silage
Improves silage quality
EU notified strains
Easy to use