rf mcfeeters, im pérez-díaz and sd johanningsmeier usda
TRANSCRIPT
RF McFeeters, IM Pérez-Díaz and SD Johanningsmeier
USDA-ARS Food Science Research Unit
Nancy Custis, Carl Gilbert, and Julius Rideout, B&G Foods, Inc.
Lisa Moeller, Janet Turner, Kelly Beene,
Dwight Bass, Chris Whitley and Doug Brock; Mount Olive Pickle Company
Waste water containing high levels of NaCl from cucumber fermentation tank yards is a continuing problem for the pickled vegetable industry.
A major reduction in waste salt could be achieved if NaCl was eliminated from the cucumber fermentation process.
Thus the objective is to ferment cucumbers in cover brines containing CaCl2 as the only salt.
And to compare firmness retention of cucumbers fermented in CaCl2 brine during subsequent storage to cucumbers fermented in brines containing NaCl.
IPD-150-12
CaCl2 fermentations were similar to NaCl fermentations • in the chemical changes caused by the fermentative
microorganisms
• and in the retention of firmness in the fermented cucumbers
• CaCl2 fermentations were also stable (in jars, in the absence of air)
However, post-fermentation pH was lower in the
presence of CaCl2 than in the presence of sodium chloride (~3.0) McFeeters RF, Pérez-Díaz IM. 2010. Fermentation of cucumbers brined with
calcium chloride instead of sodium chloride. J Food Sci 75(3):C291-C296.
IPD-151-12
Trials at B&G Foods, MD
in open top tanks
• Primary fermentation
proceeded successfully,
however a secondary
fermentation developed
after 4 months of storage
IPD-152-12
• It was observed that a key factor in doing fermentations without salt is to limit purging only to what is needed to prevent bloating, though the minimum required is not completely clear.
• Sorbic acid is effective in limiting yeast growth, but we wanted to also look at other preservatives such as benzoate and AITC to see if we had multiple options.
IPD-153-12
Aromas from the fermentation barrels were clean, although some film yeast formed on the surfaces of the cover brines.
Finally there was the question of whether post-fermentation spoilage would occur over extended periods of storage; and whether preservatives added to the cover solution before or after fermentation would prevent it.
IPD-154-12
Control Tanks Experimental Tanks
Air Purging Routine
• Initiated the morning after the tanking day
• 4 hours on and 4 hours off for 7 days
• purged on that same schedule on days 9, 11, 13 and then
stopped the purging
6% NaCl
1.1% CaCl2
0.05% acetic acid
1.1% CaCl2
0.09% potassium sorbate
starter culture (L. plantarum)
0.32% fumaric acid (3 weeks later)
IPD-155-12
CaCl2: firmness retention with time
Potassium sorbate and the minimized air purging routine: prevention of the proliferation of yeasts during fermentation and storage
Fumaric acid: to prevent the growth of spoilage associated lactic acid bacteria during storage
Removal of acetic acid: to reduce the extent and/or probability of secondary cucumber fermentation or spoilage
Starter culture: outcompete the natural microbiota
IPD-156-12
Fresh Cucumbers: 5.22 ± 0.93 Log of CFU/mL
Fresh CaCl2 Brines: 1.44 ± 1.89 Log of CFU/mL
Fresh NaCl Brines: below detection level
Inocula: 8.17 ± 0.55 Log of CFU/mL
• Preliminary data suggest that Starter Cultures used as inocula prior to reaching maximum cell density (9 Log of CFU/mL) grow faster in the commercial fermentation.
Initial LAB Counts: 5.88 ± 0.59 Log of CFU/mL
IPD-157-12
Time (days)
0 50 100 150 200 250 300
LA
B C
ou
nts
(L
og
CF
U/m
L)
2
3
4
5
6
7
8
9
CaCl2 MRS
NaCl MRS
Counts of lactic
acid bacteria
reached a
maximum after
0.75 days in the
CaCl2
fermentations as
compared to 4
days in the NaCl
fermentation.
IPD-158-12
Time (days)
0 50 100 150 200 250 300
Glu
cose
con
cen
trati
on
(m
M)
0
2
4
6
8
10
12
14
16
NaCl Fermentation
CaCl2 Fermentation
Time (days)
0 50 100 150 200 250 300
Fru
cto
se c
on
cen
trati
on
(m
M)
0
5
10
15
20
25
NaCl Fermentation
CaCl2 Fermentation
Carbohydrate utilization proceeds faster in CaCl2 fermentations as compared
to NaCl fermentations
IPD-159-12
0.25% 0.36%
Time (days)
0 50 100 150 200 250
pH
2
3
4
5
6
7
8
NaCl Fermentation
CaCl2 Fermentation
Time (days)
0 50 100 150 200 250
La
ctic
aci
d c
on
cen
trati
on
(m
M)
0
20
40
60
80
100
NaCl Fermentation
CaCl2 Fermentation
IPD-160-12
0.72%
Yeasts proliferated immediately after the primary fermentation,
presumably deriving energy from the residual carbohydrates or from
lactic acid.
Time (days)
0 50 100 150 200 250 300
Ye
as
t C
ou
nts
(L
og
CF
U/m
L)
2
3
4
5
6
7
CaCl2 YMA
NaCl YMA
IPD-161-12
Time (days)
0 50 100 150 200 250 300
So
rbic
aci
d c
on
cen
trati
on
(m
M)
0
2
4
6
8
10
12
14
16
18
NaCl Fermentation
CaCl2 Fermentation
0.18%
0.02%
Time (days)
0 50 100 150 200 250
Ace
tic a
cid
con
cen
trati
on
(m
M)
0
10
20
30
40
50
NaCl Fermentation
CaCl2 Fermentation
IPD-162-12
0.24%
0.08%
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
0 0.5 1 2 3 7 14 21 30 60 77 89 103 118 153 187
LA
B C
ou
nts
(L
og
of
CF
U/
mL
)
Time (days)
Processed
Phosphoric acid & sodium benzoate
were added
Potassium sorbate
Sodium benzoate
Fumaric acid
Phosphoric Acid
IPD-163-12
2
3
4
5
6
7
8
9
0 0.5 1 2 3 7 14 21 30 48 60 74 89 124 158
LA
B C
ou
nts
(L
og
of
CF
U/
mL
)
Time (days)
Potassium sorbate was added
Potassium sorbate &
fumaric acid were added
IPD-164-12
Counts of enterobacteriacea after
culture enrichment
2.00
2.50
3.00
3.50
4.00
4.50
5.00
5.50
6.00
6.50
Day 1 Day 3 Day 7
En
tero
ba
cte
ria
Co
un
ts (
Lo
g o
f C
FU
/m
L)
Time
Exocarp (Skin) Mesocarp Endocarp (Seed Cavity)
2.00
2.50
3.00
3.50
4.00
4.50
5.00
5.50
6.00
6.50
Day 1 Day 3 Day 7
En
tero
ba
cte
ria
Co
un
ts (
Lo
g o
f C
FU
/m
L)
Time
Exocarp (Skin) Mesocarp Endocarp (Seed Cavity)
CaCl2 fermentations NaCl fermentations
IPD-165-12
Air Purging Routine • Initiated the morning after the tanking day
• 4 hours on and 4 hours off for 7 days
• purged on that same schedule on days 9, 11, 13 and then stopped the purging
1.1% CaCl2
0.09% potassium sorbate
starter culture (L. plantarum)
2 weeks post-tanking
0.32% fumaric acid, 0.17% sodium benzoate, 200 ppm AITC
or
combinations of these preservatives
IPD-166-12
2.80
3.30
3.80
4.30
4.80
5.30
5.80
1 3 7 14 21 28 43 60 62 76 90 105 112 121
pH
Time (days)
CaCl2 with Fumaric Acid and Sodium Benzoate CaCl2 with Fumaric Acid CaCl2 and Sodium Benzoate CaCl2 CaCl2 with Fumaric Acid and AITC NaCl
IPD-167-12
0.00
20.00
40.00
60.00
80.00
100.00
120.00
1 3 7 14 21 28 43 60 62 76 90 105 112 121
La
cti
c A
cid
Co
nc
en
tra
tio
n (
mM
)
Time (days)
CaCl2 with Fumaric
Acid and Sodium
Benzoate CaCl2 with Fumaric
Acid
CaCl2 and Sodium
Benzoate
CaCl2
CaCl2 with Fumaric
Acid and AITC
NaCl
IPD-168-12
0.81%
IPD-169-12
0.00
10.00
20.00
30.00
40.00
50.00
60.00
1 3 7 14 21 28 43 60 62 76 90 105 112 121
Ac
eti
c A
cid
Co
nc
en
tra
tio
n (
mM
)
Time (days)
CaCl2 with Fumaric
Acid and Sodium
Benzoate CaCl2 with Fumaric
Acid
CaCl2 and Sodium
Benzoate
CaCl2
CaCl2 with Fumaric
Acid and AITC
NaCl
0.3%
2.4
3.4
4.4
5.4
6.4
7.4
8.4
9.4
1 3 7 14 21 28 43 60 76 90 105 112 121 154
Co
un
ts o
f la
cti
c a
cid
ba
cte
ria
(Lo
g o
f C
FU
/m
L)
Time (day)
fresh cucumbers CaCl2 and Sodium Benzoate CaCl2 NaCl
IPD-170-12
2.4
2.6
2.8
3.0
3.2
3.4
3.6
3.8
4.0
4.2
4.4
1 3 7 14 21 28 43 60 76 90 105 112 121 154
Ye
as
t C
ou
nts
(L
og
of
CF
U/
mL
)
Time (day)
fresh cucumbers
CaCl2 and Sodium Benzoate
CaCl2
NaCl
IPD-171-12
The temperature fluctuated between 68 and 90
degree F
Fumaric acid and benzoic acid concentrations
were stable thru time.
Sorbic acid concentrations were unstable thru
time.
Sodium benzoate was the most robust
preservative in preventing microbial growth.
IPD-172-12
NaCl Fermented CaCl2 Fermented
pH 3.49 ± 0.11
Calcium 22 ± 5.5 mM
Slightly darker appearance
Differences in texture
varied in independent
trials
pH 3.30 ± 0.02
Calcium 22 ± 2.8 mM
Brighter in appearance,
especially as observed in
jars
SDJ-174-12
All products were within the range of a normal hamburger dill chip
product
No significant off-flavors were associated with either treatment
It is possible to ferment cucumbers and stored them for six months in a cover brine without NaCl.
The proposed CaCl2 system has the potential
to reduce the high salt wastes from vegetable fermentations.
Hamburger dill chips prepared from calcium chloride fermentations were nearly identical in sensory quality after 6 months storage in glass jars
IPD-175-12
Disadvantages: (1) new process, uncertainty (2) freshly made fermentation cover brines cannot be stored for extended periods of time (3) need to add a preservative post-fermentation (4) need to generate large volumes of starter cultures in-house (5) water usage; however a lot of water is used today with the purpose of diluting chlorides in waste waters
IPD-176-12
Advantages: (1) reduces pollution from the disposal of high
salt brines
(2) presents an opportunity for the manufacture
of low-sodium products
(3) create opportunities for new flavors in finished
products
(4) eliminates the need to recycle cover brines
(5) no carry-over of undesired flavors, enzymes,
an high acid levels from the recycled brines
IPD-175-12
Advantages: (more)
(6) reduces cost of labor and equipment operation
(7) minimizes the need for pumps on the platforms
(8) reduces energy demands
(9) eliminates the need to store used cover brines
(10) more consistent end-product
(11) less air purging
(12) the CaCl2 containing brines can be sprayed on
landfills
IPD-176-12
Ilenys M. Pérez-Díaz, PhD Research Microbiologist
USDA-ARS Food Science Research Unit, Raleigh NC
919-513-0165
IPD-177-12
Step 6: As needed a small piece of ice from
the frozen bacterial culture is transferred
to fresh and sterile 1 ml MRS broth. Here
the MRS broth can be substituted by
cucumber juice.
Step 7: The tube containing the 1 ml cucumber juice
and the bacterial cells is incubated at 30 C for 48 hours to
encourage growth (multiplication of the bacterial cells).
Preparation of a Kosher starter culture for
vegetable fermentations
IDP-33-12 Friday, January 27th, 2012
Step 8: Jars are inoculated
and incubated for 3 days at
30°C
Step 9: A portion of the
brine from this jar is pour
off onto a 1-gallon Kosher
dill spears.
Step 11: The total content
of 4 turbid 1-gallon jars ,
is pour off into a tank of
(6000-10000 gallons).
Step 10: The 1-gallon
jar is maintained at
room temperature until
turbidity develops.