Bacteria associated with ensiling fermentation and aerobic stability of total mixed ration silage
Naoki Nishino and Chao Wang Okayama University, Japan
Minimize the risk of effluent production
Counterbalance the nutrients of by-products
Improve odors and flavors during fermentation
Stabilize microbial function and improve energy and protein utilization in the rumen
Reduce the labors associated with mixing prior to feeding
Advantages of ensiling wet by-products as TMR silage XVI I S C 2012
Characteristics of TMR silage XVI I S C 2012
More than10 ingredients including wet by-products and dry feeds
High nutritive value targeted for dairy cows (CP >160 g/kg DM, TDN 720-760 g/kg DM)
Low moisture (DM 500-600 g/kg)
Short storage usually within one month
Wet brewers grains TMR silage
(oC)
26
28
30
32
24 1 2 3 4 5 6 7 d
Tem
per
atu
re
Non-ensiled TMR TMR silage
(oC)
30
40
50
20 1 2 3 4 5 6 7 d
Resistance to aerobic deterioration XVI I S C 2012
Tem
per
atu
re
-4
-3
-2
-1
0
1
2
3
4
Pala
tab
ility
Sco
re
Non-ensiled TMR TMR silage
0 h 24 h 0 h 24 h
(Yoshihara et al. 2006)
Prevention of intake depression in summer XVI I S C 2012
Lactobacillus fermentum
Lactobacillus delbrueckii
Lactobacillus plantarum
Lactobacillus brevis
Lactobacillus buchneri
Pediococcus acidilactici
Lactobacillus plantarum
Lactobacillus brevis
Lactobacillus parvulus
67%
33%
44%
30%
20%
3%
3%
Day 56
Day 14
XVI I S C 2012
Lactobacillus delbrueckii
Lactobacillus delbrueckii
Lactobacillus brevis
Lactobacillus fermentum
Lactobacillus fermentum
Lactobacillus fermentum
Lactobacillus buchneri
M 14 56
43%
57%
Pre-ensiled TMR mixture
TMR silage
Plate-culture Culture-independent DGGE
Lactobacillus buchneri
Lactobacillus buchneri 44%
Association of L. buchneri in the ensiling process
Seasonal changes − a survey of commercial products − XVI I S C 2012
−2 − 10℃ 5 − 25℃ 20 − 30℃ 5 − 22℃
Jan−Feb Apr−May Jul −Aug Oct−Nov
50
40
30
20
10
0
40
30
20
10
0
80
60
40
20
0
Lactic acid (g/kgDM)
Acetic acid (g/kgDM)
Ethanol (g/kgDM)
−2 − 10℃ 5 − 25℃ 20 − 30℃ 5 − 22℃
XVI I S C 2012
Jan−Feb Apr−May Jul −Aug Oct−Nov
Weissella paramesenteroides Weissella paramesenteroides Lactobacillus plantarum Lactobacillus fermentum Lactobacillus brevis Pediococcus pentosaceus Lactobacillus panis Lactobacillus pontis Lactobacillus vaginalis Lactobacillus frumenti
Lactobacillus plantarum Lactobacillus fermentum
Pediococcus pentosaceus
Jan-Feb Apr-May Jul-Aug Oct-Nov
Lactobacillus panis Lactobacillus pontis Lactobacillus vaginalis Lactobacillus frumenti
Seasonal changes − a survey of commercial products −
Pre-ensiled TMR (commercial product)
5℃
15℃
25℃
35℃
Storage for 10, 30 & 90 days
5℃
15℃
25℃
35℃
5℃ 25℃ 15℃ 25℃
25℃ 25℃ 35℃
XVI I S C 2012
−2 − 10℃ 5 − 25℃ 20 − 30℃ 5 − 22℃
Jan−Feb Apr−May Jul −Aug Oct−Nov
Temperature effects − a laboratory-scale experiment −
Storage temp
Stability test
10 days 30 days 90 days
5℃ 5℃
25℃
15℃ 15℃
25℃
25℃ 25℃
35℃ 25℃
35℃
○ ○ ○
○ ○ ○
○ ○ ○
○ ○ ○
× × ○
× ○ ○
○ ○ ○
XVI I S C
2012 Warm temperature and long storage confirm stability
Days after ensiling Days after ensiling Days after ensiling
5℃ 15℃ 25℃ 35℃
0 10 30 90
80
60
40
20
0
40
30
20
10
0
0 10 30 90
40
30
20
10
0
0 10 30 90
Lactic acid (g/kgDM)
Acetic acid (g/kgDM)
Ethanol (g/kgDM)
XVI I S C 2012 Temperature effects on acetic acid and ethanol contents
Lactobacillus helveticus Lactobacillus brevis Lactobacillus sp. Lactobacillus plantarum Acinetobacter baumannii Lactobacillus sp. Tetragenococcus halophilus Lactobacillus buchneri Lactobacillus panis Lactobacillus sp. Lactobacillus panis Lactobacillus sp. Lactobacillus delbrueckii Lactobacillus pontis
XVI I S C 2012
Pre
-en
sile
d
5 o
C
15
oC
2
5 o
C
35
oC
5
oC
1
5 o
C
25
oC
3
5 o
C
5 o
C
15
oC
2
5 o
C
35
oC
10 days 30 days 90 days Co
mm
erci
al
sila
ge
Lactobacillus helveticus Lactobacillus brevis
Lactobacillus sp. Tetragenococcus halophilus Lactobacillus buchneri
Acinetobacter baumannii
Temperature effects on bacterial community
Lactobacillus helveticus Lactobacillus brevis Lactobacillus sp. Lactobacillus plantarum Acinetobacter baumannii Lactobacillus sp. Tetragenococcus halophilus Lactobacillus buchneri Lactobacillus panis Lactobacillus sp. Lactobacillus panis Lactobacillus sp. Lactobacillus delbrueckii Lactobacillus pontis
XVI I S C 2012
Pre
-en
sile
d
5 o
C
15
oC
2
5 o
C
35
oC
5
oC
1
5 o
C
25
oC
3
5 o
C
5 o
C
15
oC
2
5 o
C
35
oC
10 days 30 days 90 days Co
mm
erci
al
sila
ge
Lactobacillus brevis
Lactobacillus sp.
Lactobacillus buchneri
Lactobacillus panis Lactobacillus sp.
Lactobacillus pontis
Lactobacillus panis
Temperature effects on bacterial community
XVI I S C 2012 Conclusions
Product seasons (storage temperature) have effects on fermentation and bacterial community
Ehanol may increase in winter and acetic acid will increase in summer.
Long storage can confirm aerobic stability even at low temperature storage.
A number of bacteria have potential to inhibit spoilage
L. buchneri is not essential to support stability.
Inoculant effects and functions of L. panis and L. pontis are worth examining.