United States
Environmental Protection
Agency
Industrial Environmental Research
Laboratory
Cincinnati OH 45268
EPA-600/2-80-046
February 1980
Research and Development
Reducing
Wastewater from
Cucumber Pickling
Process by
Controlled Culture
Fermentation
PROTECTION
AGENCY
DALLAS, TEXAS
UNUOT
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development, U S Environmental
Protection Agency, have been grouped into nine series These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are
1 Environmental Health Effects Research
2 Environmental Protection Technology
3 Ecological Research
4 Environmental Monitoring
5 Socioeconomic Environmental Studies
6 Scientific and Technical Assessment Reports (STAR)
7 Interagency Energy-Environment Research and Development
8 "Special" Reports
9 Miscellaneous Reports
This report has been assigned to the ENVIRONMENTAL PROTECTION TECH-
NOLOGY series This series describes research performed to develop and dem-
onstrate instrumentation, equipment, and methodology to repair or prevent en-
vironmental degradation from point and non-point sources of pollution. This work
provides the new or improved technology required for the control and treatment
of pollution sources to meet environmental quality standards.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161
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EPA-600/2-80-046
February 1980
REDUCING WASTEWATER FROM CUCUMBER PICKLING PROCESS
BY CONTROLLED CULTURE FERMENTATION
by
Linda W. Little
Jeffrey G. Wendle
Jeffrey Davis
University of North Carolina at Chapel Hill
and
Robert M. Harrison
Samuel J. Dunn
North Carolina A & T State University
Greensboro, North Carolina
Grant No. S-804220
Project Officers
Kenneth A. Dostal
and
Harold W. Thompson
Food and Wood Products Branch
Industrial,Environmental Research Laboratory
Cincinnati, Ohio 45268
INDUSTRIAL ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI, OHIO 45268
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DISCLAIMER
This report has been reviewed by the Industrial Environmental Research
Laboratory-Cincinnati, U.S. Environmental Protection Agency, and approved
for publication. Approval does not signify that the contents necessarily
reflect the views and policies of the U.S. Environmental Protection Agency,
nor does mention of trade names or commercial products constitute endorse-
ment or recommendation for use.
ii
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FOREWORD
When energy and material resources are extracted, processed, converted,
and used, the related pollutional Impacts on our environment and even on
our health often require that new and increasingly more efficient pollution
control methods be used. The Industrial Environmental Research Laboratory -
Cincinnati (lERL-Ci) assists in developing and demonstrating new and im-
proved methodologies that will meet these needs both efficiently and econo-
mically.
This report presents an evaluation of a modified cucumber pickling
process using controlled culture fermentation as compared to the conven-
tional natural fermentation process. At commercial scale the modified pro-
cess produced a product equal to or exceeding that of the natural fermen-
tation with a significant reduction in the quantity of salt used during
fermentation. Further information on the subject can be obtained by con-
tacting the Food and Wood Products Branch, Industrial Environmental Research
Laboratory-Cincinnati.
David G. Stephan
Director
Industrial Environmental Research Laboratory
Cincinnati
iii
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ABSTRACT
On a demonstration scale, the controlled culture fermentation process
(CCF) developed by the U. S. Food Fermentation Laboratory was compared with
the conventional natural fermentation process (NF) in regard to product
quality and yield and volume and concentration of wastewaters. The experi-
ments were conducted at the Perfect Packed Products Company, Inc.,
Henderson, North Carolina. Nine 800-gal. tanks were brined. In each case,
weight of cucumbers, volume of water, and amounts of additives were recorded.
pH, acidity, salinity, and temperature were closely monitored. After
brining, brinestock quality was evaluated by a panel of experts from the
US Food Fermentation Laboratory and the Heinz Company. The brinestock
was then processed; spent brines and processing waters were collected.
Volume and wastewater characteristics (salinity, BOD, N and P forms,
residues) were determined for the waters and weight of brinestock was
determined. The cucumbers were then packed using a conventional finishing
procedure for whole dill pickles and hamburger dill chips. Yield of final
product was determined. Acceptability of the finished products was
evaluated by a panel.
Analysis of data indicates that the CCF produces a product of quality
equal to or exceeding that of NF; that a reduction of the total dissolved
solids load in the wastewaters was achieved; and that fermentation occurs
more rapidly and predictably. Under the carefully controlled conditions
of the experiment, the NF procedure produced brinestock of better quality
than that usually achieved by this process, indicating that higher yields
could be obtained with existing tankyard procedure if better control over
tank condition and over salting schedules was maintained.
Experiments on recycling of spent brines indicate that in the
coagulation-precipitation reconditioning procedures inactivation of
undesirable enzymes is due to denaturation of the enzymes at high pH rather
than to physical removal of the enzymes by flocculation. A combination of
lime and sodium hydroxide at pH's above 10 produced a clear brine with
little or no enzyme activity. Lower pH levels, or use of alum and polymers
could achieve clarification of the brine but enzyme activity remained.
Brine recycling studies also indicate that CCF spent brine cannot be reused
directly unless the brine pH is raised above the brine's characteristic
pH 3.2 - 3.5, which is too low for the desirable Lactobacilli.
NOTE: This report follows the prevailing canning industry practice of using
the international system of units in the laboratory and U.S. units in the
manufacturing operations. A table of conversion factors is included.
iv
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CONTENTS
Foreword ill
Abstract iv
Figures vi
Tables vii
Abbreviations and Symbols ix
Conversion Factors x
Acknowledgments xii
1 Introduction 1
General 1
Types of packs 1
Pickle production and the wastewater generated 1
Approaches to reduction of wastes 3
Objectives of project 4
2 Conclusions 5
3 Recommendations 6
4 Experimental Design 7
Approach 7
Materials 7
Methods for brine and wastewater analyses 9
Evaluation of quality of brinestock and finished products 12
Procedures for fermenting cucumbers 12
Experiments on brine recycling 16
5 Results and Discussion 19
Comparison of natural and controlled culture fermentations 19
Brine recycling studies 41
Economic evaluation 60
6 References 65
7 List of Publications 67
8 Appendix 68
v
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FIGURES
Number Page
1 Photograph of experimental tanks 14
2 Comparison of lactic acid production in controlled culture
and natural fermentations (three experiments) 20
3 Evaluation of hamburger chips prepared by natural fermentation
(including a commercial product) and by controlled culture
fermentation 22
4 Evaluation of whole dill pickles prepared by natural fermenta-
tion and by controlled culture fermentation 23
5 Progress of desalting in experiment 1: Salt concentration in
processing water as a function of time 35
6 Carbon dioxide accumulation in experimental tanks, experiment 1 . 37
7 Carbon dioxide accumulation in experimental tanks, experiment 2 . 38
8 Carbon dioxide accumulation in experimental tanks, experiment 3 . 39
9 Brine turbidity as a function of alum dosage, initial pH = 7 . . . 43
10 Enzyme activity as a function of ludox dosage . 49
11 Enzyme activity as a function of celite dosage 51
12 The effect of pH on the enzyme activity of a no. 2 spent brine . . 53
13 Effect of pH on enzyme activity in "spiked" brine samples .... 54
14 Enzyme activity as a function of pH for 35 °S, 0.1 lactic acid
"synthetic" brine 56
15 Enzyme activity as a function of pH synthetic brine, 35 °S,
acetic acid 57
16 Enzyme activity of a spent brine as a function of pH 58
17 Pectinase activity as a function of alum dosage in a distilled
water sample 59
vi
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TABLES
Number Page
1 Salt (NaCl) solution conversion chart 10
2 Controlled culture fermentation procedure 13
3 Natural fermentation procedure 15
4 Evaluation of brinestock from natural fermentations and from
controlled culture fermentations 21
5 Taste tests, experiment 3 - (2 cores per tank tested) 25
6 Statistical analysis of experimental results, Analysis: One-
way analysis of variance, different sample sizes 26
7 Comparison of quantity and composition of brines from natural
and controlled fermentations (expt. 1) 27
8 Comparison of quantity and composition of brines from natural
and controlled fermentations (expt. 2) 29
9 Comparison of quantity and composition of brines from natural
and controlled fermentations (expt. 3) 32
10 Summary comparison of quantity and composition of brines from
natural and controlled fermentations 34
11 CC>2 accumulation in large commercial brinings 40
12 Sodium aluminate jar test 42
13 Turbidity and enzyme activity as a function of alum dosage,
initial pH = 5 44
14 Turbidity as a function of alum dosage, initial pH = 5.45 ... 45
15 Turbidity and enzyme activity as a function of alum dosage
@ initial pH = 7 45
16 Effects of NALCO 8151 and optimum alum dosage, initial
pH = 7 46
vii
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Number Page
17 Effects of NALCO 8151 and optimum alum dosage, initial
pH = 7 46
18 Metals content in a no. 2 brine used for jar tests, compared
with those reported by Henne and Geisman (1973) 47
19 Effects of high molecular weight, anionic polymer (NALCO 7744A)
on turbidity and enzyme removal at pH = 5.6 48
20 The effects of pH on turbidity and enzyme activity 52
21 Effects of treatment on brinestock quality, recycling studies . 61
22 Progress of fermentation in recycling studies 61
vlii
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ABBREVIATIONS AND SYMBOLS
BOD, BOD,. 5-day 20°G biochemical oxygen demand
G Degrees Celsius
CCF Controlled culture fermentation
Cl Chloride
COD Chemical oxygen demand
g Gram
gal Gallon
i.d. Inside dimension
JTU Jackson Turbidity Unit
kg Kilogram
1 Liter
Ib Pound
mg Milligram
min Minute
ml Milliliter
N Nitrogen
NF Natural fermentation
o.d. Outside dimension
P Phosphorus
ppm Parts per million
S Degrees salometer
SS Suspended solids
TDS Total dissolved solids
TKN Total Kjeldahl nitrogen
TOC Total organic carbon
TSS Total suspended solids
IX
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CONVERSION FACTORS AND METRIC PREFIXES3
CONVERSION FACTORS
To convert from
to
Degree Fahrenheit (°F)
inch (in)
foot (ft)
gallon (gal)
bushel (bu)
3.8 gal/bu
grain (gr)
ounce (oz)
pound (Ib)
gallons per minute
(gal/min)
ounces per gallons
(oz/gal)
pounds per ton
(Ib/ton)
pounds per 1000 pounds
(lb/1000 Ib)
tons per year
(ton/yr)
gallons per ton
(gal/ton)
Degree Celsius (°C)
metre (m)
metre (m)
metre3 (m3)
metre (m )
0.408 m3/m3
kilogram (kg)
kilogram (kg)
kilogram (kg)
O
metre /second
(m3/s)
f\
kilogram/metre
(kg/m3)
kilogram/kilokilogram
(kg/kkg)
kilogram/kilokilogram
(kg/kkg)
kilokilogram/year
(kkg/yr)
metre /kilokilogram
(m3/kkg)
Multiply by
t°c =0.56 (t°F-32)
2.54 x 10~2
3.048 x 10"1
3.784 x 10-3
3.524 x 10~2
1.0
6.48 x 10~5
3.11 x 10~2
4.536 x 10"1
-5
-1
6.308 x 10
8.218
4.643 x 10
1.0
9.074 x 10~]
4.17 x 10~ 3
x
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To convert from
to
pounds per gallon
(Ib/gal)
pounds per 1000 gallons
(lb/1000 Ib)
cost per pound
Multiply by
cost per gallon
($/gal)
cost per 1000 gallon
($/ton)
kilogram/metre3
(kg/m3)
o
kilogram/metre
(kg/m3)
cost/kilogram
($/kg)
cost/metre3
($/m3)
cost/metre3
($/kkg)
1.1984 x 102
1.1984 x KT1
4.536 x 10"1
2.642 x 102
2.642 x 10
-1
METRIC PREFIXES
Prefix
kilo
centi
Symbol
k
Multiplication factor
10-
10
-2
Example
2 kg = 2 x 103 grams
2 cm = 2 x 10~2 metre
aStandard for Metric Practice. ANSI/ASTM Designation: E 380-766, IEEE Std
268-1976, American Society for Testing and Materials, Philadelphia,
Pennsylvania, February 1976. 37 pp.
xi
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ACKNOWLEDGMENTS
Without the cooperation and support of Perfect Packed Products Company,
Inc., Henderson, N. C., a division of Heinz Company, this project would not
have been possible. Personnel at this plant provided invaluable assistance.
Special recognition is due to George Daily, Plant Manager, Bailey Kearson,
Tankyard Manager, and Milton Alligood, Quality Control.
Mr. Thomas A. Bell and Dr. Henry Fleming, U. S. Food Fermentation Lab-
oratory, Raleigh, N. C., who participated in the development of the controlled
culture fermentation process, assisted in the project in experimental
design, evaluation of brinestock and product quality, and enzyme analysis.
Their participation in this project was in great measure responsible for
its successful completion.
Special thanks is extended to Mr. Lloyd Hontz, Vice-President, Mount
Olive Pickle Company, for supplying cucumbers for laboratory scale experi-
ments. Appreciation is also due Miles Laboratories Inc. which provided
some of the cultures used in the project.
Ms. Mablelene Smith, Program Coordinator, gave freely of her time and
contributed exceptional skills in the preparation of this manuscript for
which we are deeply grateful.
xii
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SECTION 1
INTRODUCTION
GENERAL
The cucumber pickle has long been a popular item in the American diet,
and its popularity is increasing as shown by the following per capita con-
sumption figures from the USDA Statistical Reporting Service:
Year lb/capita
1940 2.88
1950 4.62
1960 5.19
1970 7.60
1976 8.26
In terms of standard 24/303 cases, in 1976 74 million cases were sold for a
total retail sales value of $593,982,000.
TYPES OF PACKS
There are two major types of cucumber pickle packcured and freshpack.
Cured pickles undergo natural fermentation and storage in salt brines.
During the storage period the salt concentration of the brine is very high
(45-65% saturation) so the brined cucumbers must be partially desalted before
being packed in vinegar solution. Fresh-pack pickles, on the other hand,
are prepared from uncured, unfermented cucumbers, packed directly in vinegar
solution, and heat-sterilized. Currently about 40% of the annual crop is
made into freshpack products.
PICKLE PRODUCTION AND THE WASTEWATERS GENERATED
Fresh packing of cucumbers creates relatively small wastewater loads.
The two major sources of wastewater are washwaters and pasteurizer cooling
waters. These streams are typically low in BOD, COD, nutrients, and
chlorides. Due to the seasonal availability of fresh cucumbers, these fresh
pack products can only be made during the "green season."
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On the other hand, cucumbers preserved by the ancient practice of
pickling can be stored up to three years before final packing. The three
major operations in production of cured pickles are (1) brining, (2)
"processing" or freshening (desalting), and (3) finishing. In the first
stage the green cucumbers, usually unwashed, are placed in a salt brine
maintained at about 25-30 °S.* This salt concentration favors growth of
indigenous lactic acid bacteria which convert the sugars diffusing out of
the cucumbers into lactic acid, lowering the pH and further suppressing the
growth of undesirable organisms. When the fermentation is complete the pH
will be about 3.4 to 3.6 and most of the sugar will have been converted to
lactic acid. After the active fermentation period, the level is raised to
45-65 °S to further suppress bacterial activity. In the low pH, high salt
environment the cucumber tissue cures, i.e., it loses its opaque white
appearance and becomes translucent. When the brinestock is ready for
packing, it is removed from the spent brine and processed in fresh water to
lower the salt content. The spent brine and desalting (or processing) waters
are characterized by their low pH, high salt content, and high oxygen demand.
The spent brine and desalting waters usually become wastewaters. Before, or
after processing the brinestock may be subjected to various treatments such
as slicing, chipping, and dicing. Wastewater from these operations is also
characterized by high salt concentration, low pH, and high organic load.
A detailed characterization of the pollutional characteristics of spent
brines and other pickling wastewaters was conducted in a previous study
(Little, Lamb, and Horney, 1976). A typical spent brine has the following
characteristics:
Total organic carbon, mg/1 3,400
Suspended solids, mg/1 330
Total Kjeldahl nitrogen, mg/1 732
Total phosphorus, mg/1 87
Chlorides, g/1 111
pH 3.4
Spent brines represent a major source of the total load of wastewaters
generated in pickle packing. They are the major source of the salt
content.
*In the pickle industry brine strengths are expressed in terms of degrees
salometer, measured with a hydrometer calibrated in percent saturation with
respect to sodium chloride. A saturated salt solution would read 100°S.
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APPROACHES TO REDUCTION OF WASTES
Because of the difficulty and expense of removing salt from wastewaters,
a number of approaches have been proposed for reducing the quantity of salt
discharged in the final plant effluent. One approach is regeneration and
reuse of the salt brines. Regeneration may be accomplished with chemical
treatment (Popper, 1967; Geisman and Henne, 1973; Palnitkar and McFeeters,
1974; Little et al., 1976), heat treatment (McFeeters et al., 1978) or
physical treatment such as reverse osmosis or molecular filtration (Little
et al. ,1976).
Another approach is adoption of lower salometer salt storage for the
brinestock. Despite the heavy use of salt, natural fermentations have been
described as "unrestricted, heterogeneous, highly complex, and variable,"
often leading to production of defective brinestock (Etchells et al.,1973).
Lowered brinestock quality is especially apt to result from growth of yeasts
or coliform-type bacteria in the brines.
Because of the problems and unpredictability of natural fermentations,
the U.S. Food Fermentation Laboratory has developed the controlled culture
fermentation (CCF) process (Etchells et al. ,1973). This process was designed
to minimize such brinestock defects as bloaters, poor texture, and off-
tastes.
In brief, the CCF process involves removal or suppression of indigenous
bacteria, followed by heavy inoculation with the desired lactic acid
bacteria. During the resulting rapid fermentation, the sugars are rapidly
consumed and high acidity is achieved. Because of the high acidity and the
absence of undesirable organisms, the brinestock can be stored at 25 °S,
requiring about half or less of the salt commonly used. The following
calculations indicate the potential salt savings with this procedure:
At 65 °S, 16.25 Ib of salt is required for each bushel of cucumbers;
At 25 °S, only 6.18 Ib/bushel is required, a reduction of 10.07
Ib/bushel (62% reduction)
Additionally, the lower salt concentration in the brinestock means that less
salt will have to be removed by processing.
The CCF process had been extensively tested in the laboratory and in
relatively small-scale tankyard studies. However, studies on a large scale,
accompanied by assessment of the potential for reducing pollutional loads,
had not been conducted.
This project was initiated in 1975. Three tankyard experiments were
conducted during the 1976 green season. In each case, both CCF and NF
(natural fermentation) tanks were set up. The raw product was weighed
before brining and records of all additions of salt and other materials
were kept. Fermentation progress was closely monitored, After completion
of brining, quality and quantity of brinestock, spent brines, processing
water, and finished products were compared.
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OBJECTIVES OF PROJECT
This project addressed the following:
1. Demonstrate on a commercial scale the potential for reducing the
salt and water required for brining and processing cucumbers by
substitution of the controlled culture fermentation procedure for
the current natural fermentation procedure.
2. Compare product quantity and quality of processed pickles produced
by controlled culture fermentation with those produced in the
conventional natural fermentation procedure.
3. Compare waste streams (volume, oxygen demand, residues, chlorides,
N and P forms) under conventional and demonstration conditions.
4. Investigate the possibility of recycling CCF and NF brines both
directly and after treatment.
5. Collect data from both natural and controlled fermentation on
labor, chemical costs, capital costs, water use, wastewater charac-
teristics, and product quality to enable an economic analysis of
the financial feasibility of CCF.
6. Collect data for computation of a mass balance of salt (i.e.,
how much salt is retained in the product and sold, how much is
required in brining, and how much salt is wasted).
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SECTION 2
CONCLUSIONS
1. Controlled culture fermentations (CCF) proceed more rapidly and result
in higher levels of acidity than do natural fermentations (NF). CCF
fermentations were also more reproducible and produced consistently
good brinestock.
2. Under the conditions tested, the volume of wastewater produced per unit
weight of raw product was approximately the same. However, desalting
of CCF brinestock required less time.
3. Products made from CCF were equal to or superior to those produced By
NF. Whole dills were equal in taste and superior in appearance and
texture to those from NF. Hamburger chips from brinestock from the
two processes were similar.
4. Under the careful experimental procedures employed in this project,
the brinestock and finished products from NF test tanks were of higher
quality than those generally obtained on the tankyard, indicating that
more attention to salting schedules, control of leaks, etc., could
substantially reduce brinestock damage in the NF procedure.
5. Spent brine from the CCF tanks is substantially higher in acidity than
that from NF tanks. It cannot be recycled directly because the low pH
depresses growth of the desired lactobacilli. Adjustment to near
neutral should be the only pretreatment necessary.
6. In the high pH procedure for regenerating spent brines, the removal of
pectinase activity is due to denaturation of the enzyme by the elevated
pH rather than to removal of the enzyme by coagulation-precipitation.
Clarification of the brines by coagulation at neutral pH's failed to
eliminate pectinase activity.
7. As expected, substantially less salt was required for CCF than for NF
brinings; thus the salt concentration in the spent brines was lower.
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SECTION 3
RECOMMENDATIONS
1. Adoption of the CCF procedure by pickle companies offers the following
advantages:
(1) Reduction of salt requirements
(2) Less salt to be discharged as a waste
(3) More rapid and consistent fermentation
(4) Quicker desalting of brinestock per volume of processing water
(or if desired, lower volume of processing water per a given
desalting time)
(5) Consistent production of brinestock equal or superior to that from
NF
(6) Less loss of brinestock due to bloater formation
2. If CCF is not adopted, the NF process can be managed to produce a much
better quality and quantity of brinestock than is usually the case
by careful attention to salting schedules and improved housekeeping
practices.
3. CCF brines should be recycled after appropriate treatment to adjust the
acidity and pH.
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SECTION 4
EXPERIMENTAL DESIGN
APPROACH
The work plan for this project was directed to three major areas:
1. Large-scale demonstration of controlled culture fermentation
(CCF) and comparison with conventional natural fermentation (NF).
2. Characterization of wastewaters generated.
3. Laboratory studies on recycling spent brines.
The large-scale demonstration was conducted at Perfect Packed Products,
Inc., (PPP) Henderson, N. C., using 800-gallon tanks. Size 3 cucumbers,
the size most commonly used for hamburger chips and whole dills, were used.
PPP supplied cucumbers, water, salt, and acetic acid, as well as labor for
tanking, heading, routine monitoring of fermentation, untanking, processing,
and packing. Three experiments were conducted. In each experiment, half the
tanks were brined conventionally and the other half according to the CCF
process. Progress of fermentations was closely monitored by PPP, North
Carolina Agricultural and Technical State University (A&T), and University
of North Carolina at Chapel Hill (UNC-CH) personnel.
After brining, brinestock quality and quantity were determined in
accordance with procedures developed by the U.S. Food Fermentation Labora-
tory. The brinestock was then processed into hamburger dill chips and
whole dills.
Spent brines and desalting waters were collected for determination
of quantity and quality. A&T University personnel determined the concen-
tration of BOD5, COD, TDS, TSS, TKN, TP, and other wastewater parameters.
These brines were also used in brine recycling experiments.
After packing and storage, the quality of the finished products was
evaluated by a taste panel.
MATERIALS
The raw materials used in this project were, where possible, those in
general use on the commercial scale, since a major goal of the project was
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to compare feasibility of natural and controlled culture fermentation pro-
cesses under actual tankyard conditions.
Cucumbers
Fresh cucumbers were obtained from those received by Perfect Packed
Products, Inc. They were graded by PPP routinely into three to four major
size categories. Size 3 (1^-2 inches in diameter) cucumbers were used for
all experiments as it was felt that the large size would be the most likely
to bloat and would thus provide the most stringent test of the CCF procedure.
Salt
The salt used was that in common use at the plant. It was rock salt of
a grade suitable for brf.nirg food products.
Cultures
Cultures of Lactobacillus plantarum were employed in controlled culture
fermentation. L. plantarum, an exceptionally acid-tolerant organism capable
of rapid fermentation, has been recommended for the CCF procedure (Etchells
et. al., 1973). Starter cultures were obtained from Chr. Hansen's Labora-
tory, Inc., Milwaukee, Wisconsin, and from Miles Laboratories, Inc., Madison,
Wisconsin. L. plantarum is homofermentative, producing primarily lactic
acid and a small amount of CC^. It tolerates salt levels as high as 26-28 °S
and pH as low as pH 3.2. Cultures were shipped in dry ice and stored at
-70 &C.
Inert Gas
Compressed nitrogen gas (Air Products Co.) was used to purge controlled
culture tanks.
Tanks
Tanks were fiberglass coated wooden tanks of 800-gallon capacity,
80"-83" I. D. and 37 3/4" tall. They were fitted with stainless steel
sample ports and drains. The tanks were headed using plastic netting and
widely spaced boards drilled with holes to facilitate gas escape.
Sparging Apparatus for CCF Tanks
Each CCF tank was provided with equipment through which nitrogen gas
could be introduced to sweep CCL out of the brine. The sparging equipment
was located at the bottom of the tank and consisted of a single coil (3.5'
in diameter) of perforated plastic tubing (polyvinyl chloride tubing, Hi-Mol,
Carlon Products, Wilton, Conn.), 1.3 cm 0. D. In each coil, 12 equally spaced
perforations (1/64" I. D.) were made with a special drill and bit. The two
ends of the coil were attached to a tube through which compressed nitrogen
gas was supplied. The coil was attached to a plywood board which was
weighted down with bricks coated in parafin. The whole unit was placed in the
bottom of the tank prior to filling. The gas flow rate was controlled with
8
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a rotameter. Initial flow rate during the early rapid fermentation was
2400 ml/min (3 ml/min/gal). Flow rate was decreased when CCL levels
remained consistently low ( < 15 mg CCWlOO ml) for two consecutive readings.
Flow rate was decreased stepwise (2000 ml/min, 1600 ml/min), then maintained
at. 800 ml/min until fermentation was completed.
METHODS FOR BRINE AND WASTEWATER ANALYSES
Acidity
Determination of percent titratable acidity (expressed as lactic acid)
was performed as follows: (1) pipet 10 ml of brine sample into titration
flask, (2) add two drops of phenolphthalein indicator, (3) titrate with 0.1N
NaOH to a permanent pink end point, (4) multiply the ml of titrant required
by the factor0.09 to give percent lactic acid by volume.
ml 0.1N NaOH x 0.09 x 100 . . , , ..
10 ml brine sample = Percent lactlc acld by V°lume
This determination was specified by Mr. T. A. Bell of the U.S. Food Fermen-
tation Laboratory and is in common use in the pickle industry.
£H
For the plant studies, pH of samples was measured electrometrically with
a Fisher Accumet pH Meter standardized with two buffers at pH's bracketing
the pH of the test sample (APHA et al., 1976, Method 424). For laboratory
recycling studies, Leeds and Northrup or Beckman pH meters were employed.
Salometer
In the pickle industry brine strengths are expressed in terms of degrees
salometer ( S), as measured with a hydrometer calibrated in percent satura-
tion 100 °S. For this project, salometer readings were performed with a com-
mon salometer routinely used on the tankyard, A salt solution conversion
chart is shown in Table 1 .
Chloride
Chloride determinations were performed with a specific ion probe
(Orion) and a Fisher Accumet pH meter, according to the directions provided
by Orion.
Turbidity
In laboratory recycling studies, turbidity was determined with the aid
of a commercial turbidimeter (Each, Model 2100) using directions supplied
by the manufacturer. This procedure was consistent with Method 214A
(APHA et al., 1976).
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K
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Metals
In laboratory recycling studies, metals were measured by atomic absorp-
tion spectrophotometry. Brine samples were collected in acid-washed glass-
ware, diluted with deionized water, and acidified with concentrated nitric
acid. Analyses were performed by the UNC-CH Limnology Laboratory using a
Perkin-Elmer Model 303 atomic absorption spectrophotometer according to the
manufacturer's specifications as described in the manual "Analytical
Methods for Atomic Absorption Spectrophotometry."
Chemical Oxygen Demand (COD)
COD was measured by the dichromate reflux method (APHA et al., 1976,
Method 508) using appropriate addition of mecuric sulfate to compensate for
chloride interference.
Biochemical Oxygen Demand (BOD)
Five-day BOD was determined according to APHA et al. (1976), Method 507.
To ensure a sufficient concentration of microorganisms in the sample, the
normal flora in the sample were supplemented by addition of seed from a
municipal wastewater source.
Residue
Residues were determined according to APHA et al. (1976). Total sus-
pended solids (SS) were determined by Gooch crucible filtration followed by
drying at 103 °C and weighing of the residue (Method 208D). Total dissolved
solids (TDS) were measured gravimetrically after evaporation at 103 °C
(Method 208C).
Carbon Dioxide
Carbon dioxide was estimated by a modification of the Harleco procedure
(Fleming et al., 1974), using the Harleco C02 Apparatus.
Phosphorus
Total phosphorus (TP) measurements were performed according to the
automated ascorbic acid method (EPA, 1974), using a Technicon Autoanalyzer.
All standards and samples were run in duplicate. Prior to analysis, brines
were digested by addition of ammonium persulfate and concentrated sulfuric
acid, followed by autoclaving.
Nitrogen
Total Kjeldahl nitrogen determinations were performed according to EPA
procedures,
-------
EVALUATION OF QUALITY OF BRINESTOCK AND FINISHED PRODUCTS
Sample Procedure for Brinestock
Representative samples of brinestock were obtained by passing a lucite
cylinder (1 ft I. D.) to the bottom of the tank and removing all the brine-
stock therein by netting. Two cores were removed from each tank.
Evaluation of Brinestock for Bloater Damage
Brinestock cucumbers were cut longitudinally and examined for balloons
honeycomb, lens, and other types of bloaters (Etchells et al., 1968). Sub-
jective evaluations were based on type and severity of the defect (Fleming
et al., 1973a).
Brinestock Texture
For each core, firmness of 20 pickles was measured by the USDA Fruit
Pressure Tester with 5/16 inch tip. Firmness is expressed in-terms of
pounds resistance of the pickle. A rating of 18 and above indicates a very
firm pickle; 14-17, firm; 11-13, inferior; 5-10, soft; and below 5, mushy.
A firm pickle is considered desirable.
Evaluation of Finished Products
Experimental brinestock was processed and packed as hamburger dill
chips or whole dills in 5-gal plastic pails. After storage, selected pails
were coded and sampled for evaluation by a taste panel consisting of
project personnel, industry personnel, and U.S. Food Fermentation Laboratory
personnel. The products were rated on a 10-point scale for appearance,
taste, and texture.
PROCEDURES FOR FERMENTING CUCUMBERS
Controlled Culture Fermentation Procedure
The CCF procedure was that suggested by Etchells et al. (1973). It is
outlined in Table 2. In-tank shrinking was employed. Chlorination was
achieved by addition of calcium hypochlorite (Lo-Bax , Olin Corporation).
The only exception to the published procedure was omission of the second
chlorination 10-12 hours before inoculation. It was assumed that suffi-
cient sanitizing was provided by the washing and initial chlorination
and that further chlorination after the cucumbers had been soaking in
the brine might cause production of undesirable chlorinated organic
compounds.
Natural Fermentation Procedure
The NF procedure was that commonly used by the pickle company. It is
outlined in Table 3.
12
-------
TABLE 2. CONTROLLED CULTURE FERMENTATION PROCEDURE
(Adapted from Etchells et _al^., 1973)
1. Raw product receiving
Visual inspection. Grade out moldy
and diseased cucumbers.
2. Washing
Remove field soil with reel-type
washer.
3. In-tank
4. Coveirbrine
Covering and heading
Acidification and
circulation
7. Salt additions
8. Acetate addition
Place in clean sanitized tank contain-
ing a 10-12 inch deep cushion of
25 °S brine containing about 20 mg/
liter C12. Use a 65:35 (wt/wt)
ratio of cucumbers to brine.
Add chlorinated 25 °S brine to a
level about 4-6 inches above the
headboard. Circulate tank to allow
shrinkage of cucumbers.
Head with plastic netting and widely
spaced boards with holes for gas
escape.
Acidify with acetic acid (vinegar) at
the rate of 6 ml of glacial acetic
acid per gallon of brined material.
Add required salt to equalize at 25 °S.
Add 0.5% sodium acetate (18.8 g/gal)
about 2-3 hours before culture
addition.
9. Culture addition
Add lactic acid bacteria, Lactobacillus
plantarum, 2-3 hours after acetate
addition.
10. Purging action
As soon as the tank is headed, brined,
CO
Test
and acidified, purge the dissolved ^w~
from the brine with N2 gas
brine for C02, sugar, and acidity.
Discontinue purging when fermentation
is complete.
(continued)
13
-------
TABLE 2 (continued)
11. Maintaining brine strength -
12. Quality of stock
Hold at 25 °S by additions of salt
as necessary.
Examine brinestock for firmness,
bloaters, color, and cure.
Figure 1. Photograph of experimental tanks.
14
-------
TABLE 3 . NATURAL FEEMENTATION PROCEDURE
Raw product receiving
In-tank
3. Covering and heading
Salt additions
Increasing brine strength
Quality of stock
Visual inspection. Grade out moldy
and diseased cucumbers.
Place cucumbers in clean tank
containing a 12" cushion of 30 °S
brine. Use a 65:35 (wt) ratio of
cucumbers to brine.
Add 30 °S cover brine. Circulate to
allow cucumbers to shrink. Head
with plastic netting and widely
spaced boards with holes for gas
escape.
Add 1.25 Ib. salt per bushel of
cucumbers on head of tank after
covering with 30 °S brine. On the
second morning, check salt readings
at bottom and top of tank. If
readings are under 25 °S, add second
day salt addition of 1 Ib./bu. If
readings are ^_ 25 °S, recheck
salometer at end of day. If reading
is <25 °S, add required salt for
second addition. On the third morning,
check salt readings at top and bottom
of tanks. If <25 °S, add 0.5 Ib.
salt/bu. If >25 °S, recheck in
afternoon and adjust. Check on a
daily basis until fermentation is
complete. Adjust as necessary to
maintain 25 °S.
When 0.6% lactic acid acidity has
been achieved, start feeder salt.
Raise tanks ~3 °S weekly by addition
on the head of 0.6 Ib. salt/bu.
Continue addition to 40 °S, or to
45 °S if tanks will be held over the
winter.
Examine brinestock for firmness,
bloaters, color, and cure.
15
-------
EXPERIMENTS ON BRINE RECYCLING
A major consideration in brine recycling is the possibility of carryover
of pectinases, the enzymes responsible for softening of brinestock. A
secondary consideration is the carryover of suspended solids which represent
microorganisms and debris. Recycling studies were chiefly addressed to
removal of pectinases and turbidity from the brines. An additional study
involved actual recycling, on a bench-scale, of brine collected in the
demonstration phase of the project. Much of the work on recycling has been
presented as a master's report (Wendle, 1977).
The laboratory investigations involved three major experimental
procedures: (1) jar testing to determine feasibility of enzyme and turbidity
removal by coagulation-sedimentation processes, (2) analysis of enzyme
activity in treated and untreated brines to determine the extent of the
enzyme problem and to evaluate success of treatment, (3) acidity titrations
with real and synthesized brines to predict the amount of alkali needed
for pH adjustment and to determine fate of metals during brine clarification.
Jar Test Methods
Jar tests were conducted with the aid of a. six-paddle gang stirrer, using
200 ml aliquots of brine in 250 ml beakers. After addition of coagulants
or coagulant aids, the samples were flash-mixed at 100 rpm for 30 seconds.
The samples were then flocculated at 30 rpm for 30 minutes. While jar
tests generally allow for settling of 30 min to 1 hr, settling time was
not considered critical in these experiments since full-scale treatment
of brine at pickle plants is likely to be a batch operation. Consequently,
sedimentation was allowed to continue until floe had fully settled (a
minimum of 30 min) or for a maximum of 24 hr. Supernatant was removed with
a pipette. In most cases, jar tests were preceded by preliminary tests in
which visual analysis was sufficient to determine the potential of chemical
additions to provide clarification at a reasonable dosage and thus indicate
if further investigation was warranted.
A slight variation of the above procedure was used when evaluating
coagulation by pH adjustment. Because the spent brines contained such metal
species as aluminum, calcium, and magnesium, the corresponding metal precipi-
tates formed at pH levels above pH 7. In investigations of this effect, pH
was adjusted with sodium hydroxide (NaOH) or with a lime slurry (CaO and
water) during rapid mixing on a magnetic stirrer. When the sample reached
the desired pH, it was placed on the gang stirrer, flocculated at 30 rpm
for 30 min, and allowed to settle for approximately an hour.
Sample brines were obtained from actual spent brines collected from the
tankyard of Perfect Packed Products. Spent brines from brining Number 2
size cucumbers were used, since this size is fairly small and thus more
likely to be associated with pectinase enzyme. However, in most cases the
brines did not contain significant pectinase enzyme activity. Therefore,
for the purposes of experimentation, the brines were "spiked" with Pectinol,
a commercial polygalacturonase available from Rohm & Haas. The product,
supplied in powder form, was preserved by storage in a freezer. For
16
-------
experiments, a stock solution was prepared by dissolving the powder in dis-
tilled water; the solution was preserved by refrigeration and addition of
toluene.
A variety of coagulants and coagulant aids were tested:
(1) Sodium aluminate. Both reagent grade sodium aluminate and a
commercial liquid sodium aluminate preparation (Nalco 2) were evaluated.
Stock solutions of each were prepared by dilution with distilled water.
(2) Aluminum sulfate. A stock solution was prepared from Al£(804)3.
18 H20 to Rive a concentration of 100 g/1.
(3) Clay. Nalco 8151, a slightly anionic bentonite supplied in
solution,was added directly as obtained.
(4) Polymers. Polymers evaluated included Nalco 7144 A and Dow A-21,
high molecular weight anionic polymers, and Dow C-31, a high molecular
weight cationic polymer.
(5) Sodium hydroxide. Stock solutions were prepared from reagent
grade NaOH and distilled water.
(6) Lime. Lime was added as a slurry prepared from reagent grade CaO
and distilled water.
Enzyme Activity Analysis
Enzyme analyses were performed according to U.S. Agricultural Research
Service recommendations (Bell Etchells, and Jones, 1955). This procedure
is based on indirect determination of enzyme activity via measurement of the
change in viscosity of a 1.2% sodium polypectate solution. In the presence
of pectinase enzyme, the viscosity is decreased in proportion to the amount
of enzyme activity present.
Samples for enzyme analysis were collected by pipette, placed in 20 ml
screw-top test tubes, preserved by addition of 1-3 drops of toluene, and
refrigerated until dialysis.
Dialysis was performed to reduce interference from chlorides, which
cause gelling of pectate solution and thus interfere with viscosity changes
(Bell, Etchells, and Jones, 1955). Dialyses were performed with seamless
cellulose dialysis tubing (.Fisher 8-667C 1974). Samples were immersed
for 3 hr in a continuous-flow tapwater bath. The bath was then drained and
refilled with distilled water, in which the sairples were immersed for a
minimum of an hour. Samples were then transferred to screw-top test tubes
and refrigerated until analysis.
Enzyme activity was measured in an Ostwald-Fenske viscometer as
specified. Loss of viscosity was calculated and pectinase enzyme activity
was determined as specified.
17
-------
The recommended procedure was modified in that only 20 hr viscosity
determinations were made, whereas, the protocol states that if a 50% loss in
viscosity does not occur within the first 20 hr, then a 44 hr determination
should be made. Within the scope of this project, it was felt that the
additional time and effort needed for 44 hr tests was unwarranted, since
preliminary tests indicated that the 20 hr tests were somewhat conservative
and yielded the same or slightly higher readings than the 44 hr tests.
Acidity Titrations
To determine acidity and buffer capacity, acidity titrations were per-
formed using 50 ml sample aliquots of spent brines. Titrations were generally
completed at pH 11, the pH generally recommended for coagulation with bases
(Little et al., 1976). Further titrations were conducted on synthetic
brines which were prepared to simulate spent brines in terms of acidity and
aluminum, magnesium, and salt concentration. Titration curves generated
with these synthetic brines were compared with those from actual brines to
help identify the processes responsible for precipitation. Sodium hydroxide
solution (0.1 N, Fisher Scientific Co.) was used for all acidity titrations.
Synthetic brines were prepared with distilled water and the following
reagent grade chemicals: sodium chloride, concentrated lactic acid, aluminum
sulfate, magnesium chloride. The resulting brine was 35 °S with an acidity
of 0.1 M as lactic acid, containing 70 mg and 500 mg A12(804)3- 18 H2° Per
liter.
Analytical Methods
Turbidity was measured with the aid of a Hach turbidimeter, Model 2100.
pH was determined electrometrically with a Leeds & Northrup or Beckman pH
meter.
For metal analyses, samples were collected in acid-washed glassware,
diluted with deionized water, and acidified with concentrated nitric acid.
Analyses were performed with the aid of a Perkin-Elmer atomic absorption
spectrophotometer, Model 303, according to the manufacturer's instructions
as described in "Analytical Methods for Atomic Absorption Spectrophotometry."
Bench-scale Recycling Studies in 5~Gal Pails
Limited recycling studies with spent brines were conducted. Fresh cu-
cumbers (no. 3) were obtained from a nearby commercial pickling plant. They
were packed into 5~gal plastic pails and "headed" with a rigid plastic grid.
The brinings were conducted in the laboratory at 19-22"G; throughout the test
the pails were held under constant ultraviolet lighting from a bank of germi-
cidal lamps, in order to minimize surface growths. Six pails were brined
according to the NF procedure (Table 3). Tw° pails (l and 2) served as con-
trols and received the usual new brine (30 °S). Pails 3 and ^ were brined with
NF spent brine (Tank 6, Expt. 3) diluted to 30°S; the brine for pail 3 was
treated with the high pH method before recycle, while pail 4 received untreated
brine. Pails 5 and 6 were brined with undiluted, untreated brine from GCF
fermentation (Tank 5i Expt. 3)i initial salometer was 25°S. Temperature,
salometer, acidity, and sugar content were monitored; brinestock was evaluated
at completion of the study. Sugar in brines was measured with a kit(Diastix ).
18
-------
SECTION 5
RESULTS AND DISCUSSION
COMPARISON OF NATURAL AND CONTROLLED CULTURE FERMENTATIONS
During this project, three plant-scale experiments were conducted. In
experiment I, initiated June 23, 1976, two tanks were brined, one by natural
fermentation (NF) and one by controlled culture fermentation (CCF). In
experiment II, initiated July 10, 1976, four tanks were brined, two by NF
and two by CCF; in experiment III, August 3, 1976, one by NF and two by CCF.
Day to day measurements on the brines are shown in the Appendix in
tabular and graphic form. Figure 2 shows the progress of fermentation in
the tanks as indicated by the production of acidity. In the case of dupli-
cate tanks, the average value is shown. The median afternoon brine tempera-
tures were 80.5 °F (expt. I); 84.2 °F (expt. II); and 84.8 °F (expt. III).
The initial acidity was higher in the CCF tanks due to addition of acetic
acid, as directed in the CCF procedure. Note that rate of acid production
was generally more rapid in CCF tanks and that higher final acidities were
attained in these tanks. This indicates the desired high activity of the
bacteria used for the inoculum, and it also indicates that fermentation time
can be shortened by use of the CCF procedure.
In each case, the brinestock produced in the vats was evaluated for
texture and for defects, especially those due to bloating. Overall accept-
ability and determination of usable brdnestock was computed by two different
numerical systems, one developed by Fleming et al. (1977) and one developed
by S. D. Rubin of Perfect Packed Products. A summary of the evaluation
is shown in Table 4, which indicates that CCF brinestock typically had a
lower bloater index. It is also apparent that the quality of the CCF
brinestock was much more consistent and predictable than that of NF brine-
stock, despite the fact that the NF experimental tanks received much more
attention than would a tank in the typical tankyard.
The brinestock from the experimental tanks was processed into hamburger
dill chips and into whole dill pickles. Sample packs from the first two
experiments were evaluated by a six-member panel made up of USDA, Perfect
Packed Products, and A & T personnel. Figure 3 indicates that in terms of
appearance, taste, and texture, chips prepared by NF and CCF brinings were
similar in quality and compared well with those produced commercially.
Figure 4 indicates that appearance and texture of whole dills prepared from
CCF were better than those from NF, while taste was similar.
19
-------
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TABLE 4. EVALUATION OF BRINESTOCK FROM NATURAL FERMENTATIONS
AND FROM CONTROLLED CULTURE FERMENTATIONS
Natural fermentations
Examination
for freedom from defects
Pressure test
Expt. #Tested Average (Ib) # Examined
I
II
III
Average
Controlled
I
II
III
Average
40
40
40
40
culture
40
40
40
40
40
16.7
20.0
17.0
20.5
18.6
fermentations
17.8
19.5
18.5
21.5
22.0
19.9
200
200
200
160
200
200
190
200
200
Estimated yield (
82.4
97.0
96.0
78.0
88.4
97.5
93.0
92.5
97.5
95.5
95.2
Bloater
:%)" index2
13.8
1.1
1.6
17.2
8.4
2.1
1.9
1.2
1.0
2.3
1.7
Using system devised by S. D. Rubin, Perfect Packed Products, Inc.
2Fleming et al. (1977)
21
-------
Commercial
Natural
10
Appearance
Average ratings:
NF
GGF
Taste
Appearance
8.2
8.3
Texture
Taste
7.2
Texture
7.6
FIGURE 3. Evaluation of hamburger chips prepared by natural fermentation
(Including a commercial product) and by controlled culture
fermentation.
22
-------
Natural
Appearance
Average ratings;
NP
GGF
Taste
Texture
Appearance
7.0
8.2
Taste
6.3
6.1
Texture
6.9
7.8
FIGURE 4. Evaluation of whole dill pickles prepared by natural
fermentation and by controlled culture fermentation.
23
-------
Taste tests were also conducted on the products prepared from brinestock
in experiment III. In this case, a five-member panel judged the hamburger
chips and a four-member panel judged the whole dill pickles. As shown in
Table 5, CCF chips were markedly superior in appearance and somewhat
superior in texture and taste. CCF whole dills were similar in appearance
to the NF product and markedly superior in texture, taste, and overall
acceptability. The NF tank in this experiment was the most difficult to
manage of all the NF tanks in the project. Difficulties were encountered in
keeping down yeast growth. The tank showed a more noticeable lag before
production of lactic acid (Figure 2). Softening enzyme activity was the
highest encountered in the experiments.
These results and observations served to further confirm the predict-
ability of the CCF procedure, in which the brine is initially "swamped" with
a high concentration of the desired organisms, compared to the NF procedure
in which one must depend to a large extent on luck in getting a sufficient
number of acidformers to initiate the fermentation.
Experimental results of tests of product quality from the three experi-
ments were pooled and subjected to statistical analysis to determine if
noted differences could be considered significant based on the available
amount of information. Results are summarized in Table 6. Overall, CCF
products were significantly superior to NF products in terms of texture and
acceptability.
Quantity and composition of brines from NF and CCF fermentations were
extensively monitored. Results are presented in Table 7 (expt. I), Table 8
(expt. II), and Table 9 (expt. III). They are summarized in Table 10, which
indicates the similarity of the processes in terms of amount of spent brine
generated, amount of process water generated, and BOD, COD, TKN, and TP
loads in wastewaters. As expected, TDS and TSS levels were significantly
greater in the NF brinings (Table 6). Unexpectedly, the amount of wastewater
generated in the two desalting processes did not differ significantly
(Table 6). This can be partly explained by failure to sufficiently desalt
the NF brinestock in the first experiment. In addition, it was observed
that plant personnel tended to vary duration of desalting, rather than
volume of desalting water, in response to the amount of salt left in the
brinestock. Based on plant priorities, the CCF procedure could offer
reduced desalting time per given volume of water or reduced volume of
water for a given desalting time (Figure 5).
The large significant difference in salt loading in the wastewaters from
NF and CCF brinings would have been even greater if the tanks were held for
longer periods and the salt addition to NF tanks was continued, according
to the usual practice, to a final level of 45-65 °S.
ATI examination of the data collected during the active fermentation
periods indicates that the improvement in brinestock quality noted in CCF
tanks might not be due simply to the presence of inoculum but might also
be in part due to the use of sparging. Fleming et al. (1973), noting that
bloater damage causes "serious economic losses to the pickle industry,"
reported that dissolved CO- concentration in brines is directly related to
24
-------
TABLE 5. TASTE TESTS, EXPERIMENT 3 - (2 CORES PER
TANK TESTED)*
CCF NF
Type A ₯
Whole
Appearance 7.5 7.9 7,9
Texture 7.6 7.9 6.0
Taste 5.9 7.0 5.4
Overall 6.9 7.4 5.4
Chips
Appearance 8.2 8.2 7,7
Texture 8.1 8.1 7.9
Taste 6.2 6.6 6.0
Overall 7.0 7.7 7.2
*A five-member taste panel evaluated chips; a four-member panel evaluated
whole pickles.
25
-------
TABLE 6 . STATISTICAL ANALYSIS OF EXPERIMENTAL RESULTS,
ANALYSIS: ONE-WAY ANALYSIS OF VARIANCE, DIFFERENT SAMPLE SIZES
Parameter
1) Waste waters
(spent brines
and desalting
waters)
a) Salt
b) Total sus-
pended solids
c) Volume of
wastewaters
2) Product quality
a) Pressure test
of brine-
stock
b) Texture of
whole finish-
ed product
c) Appearance of
whole finish-
ed product
d) Overall
acceptability
of whole
finished
product
Units
kg/ton of
raw product
g/ton of raw
product
gal/ton of
raw product
Ib pressure
1-10 scale
1-10 scale
1-10 scale
Mean Value
NF
105
665
279
18.6
6.6
7.2
6.0
CCF
66
481
267
19.9
7.8
8.0
7.1
a
0.005
0.05
0.05
0.05
0.005
0.05
0.01
Result
NF $ CCF
NF # CCF
NF = CCF
NF = CCF
NF ^ CCF
NF = CCF
NF + CCF
26
-------
TABLE 7 . COMPARISON OF QUANTITY AND COMPOSITION
OF BRINES FROM NATURAL AND CONTROLLED FERMENTATIONS (EXPT. 1)
Parameters
No . tanks
Cucumbers tanked, bushels
Cucumbers tanked, tons
Final Salometer
Spent brine
Volume, gal.
BOD5, mg/1
COD, mg/1
COD: BOD
TSS, mg/1
TDS, g/1
TKN, mg/1
NH3-N, mg/1
TP, mg/1
PH
Turbidity, ppm as Si02
Process (Desalting) Water
Volume, gal.
BOD5, mg/1
COD, mg/1
NF
1
72
1.80
45
364
5,000
11,160
2.2
580
67.5
425
70
93
3.5
600
182
2,700
6,475
CCF
1
74
1.85
29
284
7,800
16,000
2.1
685
48.3
418
45
84
3.8
400
178
3,300
8,330
(continued)
27
-------
Gal.
Gal.
Gal.
Gal.
BOD,
BOD,
COD,
COD,
TDS,
IDS,
TSS,
TSS,
TKN,
TO,
TP,
TP,
TABLE 7
Parameters
COD: BOD
TSS, mg/1
TDS, g/1
TKN, mg/1
NH3-N, mg/1
TP, mg/1
PH
Turbidity, ppm as SiO,
spent brine /ton tanked
spent brine/bu. tanked
process water/ton tanked
process water/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
(continued)
NF
2.4
42
48.2
200
26
32
3.2
425
202
5.05
101
2.53
4,800
120
11,040
276
70,000
1,750
600
15
404
10.1
80
2.0
CCF
2.5
49
27.4
230
9
30
3.2
750
154
3.84
96
2.41
5,600
140
12,320
308
37,600
940
560
14
328
8.2
60
1.5
28
-------
TABLE 8. COMPARISON OF QUANTITY AND COMPOSITION
OF BRINES FROM NATURAL AND CONTROLLED FERMENTATION (EXPT. 2)
Parameters
Cucumbers tanked, bu.
Cucumbers tanked, tons
Final Salometer
Spent brine
Vol., gal
BOD5, mg/1
TSS, mg/1
TDS, g/1
TKN, mg/1
TP, mg/1
pH
Turbidity, ppm as Si02
First process water
Vol., gal
BOD5, mg/1
COD, mg/1
TSS, mg/1
TDS, g/1
NF-1
80
2.0
45
297
7,200
400
152
555
122
3.8
450
52
7,200
3,820
195
63
NF-2
80
2.0
47
234
6,600
550
138
750
103
3,5
475
52
2,600
-
125
170
CCF-1
80
2.0
30
275
8,800
355
92
450
105
3,8
500
364
3,000
-
415
39
CCF-2
80
2.0
28
288
8,800
380
121
855
108
3.8
500
295
3,200
-
630
39
(continued)
29
-------
TABLE 8 (continued)
Parameters
TKN, mg/1
TP, mg/1
PH
Turbidity, ppm as S102
NF-1
245
41
3.4
350
NF-2
223
36
3.3
150
CCF-1
300
29
3.6
475
CCF-2
238
30
3.5
525
Second process water
Gal.
Gal.
Gal.
Gal.
BOD,
BOD,
TDS,
IDS,
TSS,
TSS,
Vol. gal
BOD5, mg/1
TSS, mg/1
TDS, g/1
TKN, mg/1
TP, mg/1
PH
Turbidity, ppm as Si02
spent brine /ton tanked
spent brine/bu. tanked
process water/ton tanked
process water/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
241
4,000
760
66
293
45
3.4
475
148
3.7
146
3.6
7,280
182
120,000
3,000
600
15
245
3,200
780
76
298
42
3.3
500
117
2.9
148
3.7
10,160
127
110,000
2,750
600
15
-
138
3.4
182
4.6
6,800
170
75,200
1,880
480
12
-
144
3.6
148
3.7
6,600
165
84,800
2,120
560
14
(continued)
30
-------
TABLE 8 (continued)
Parameters NF-1 NF-2 CCF-1 CCF-2
TKN, g/ton tanked 472 488 440 600
TKN, g/bu. tanked 11.8 12.2 11.0 15.0
TP, g/ton tanked 93 69 75 76
TP, g/bu. tanked 2.3 1.7 1.9 1.9
31
-------
TABLE 9. COMPARISON OF QUANTITY AND COMPOSITION
OF BRINES FROM NATURAL AND CONTROLLED FERMENTATIONS (EXPT. 3)
Parameters
Cucumbers tanked , bu .
Cucumbers tanked, tons
Final Salome ter
Spent brine
Vol., gal
BOD5, mg/1
COD, mg/1
COD: BOD
TSS, mg/1
TDS, g/1
TKN, mg/1
TP, mg/1
PH
Turbidity, as ppm Si02
Process water
Vol., gal
BOD5, mg/1
COD, mg/1
COD: BOD
TSS, mg/1
NF
80
2
37
312
8,400
16,630
2.0
410
120
495
118
3.3
250
194
3,400
10,925
3.2
1,680
CCF
80
2
33
273
11,100
20,115
1.8
335
102
495
110
3.4
250
196
6,100
13,225
2.2
740
CCF
80
2
24
270
9,400
18,650
2.0
320
80
453
105
3.5
300
204
4,900
11,625
2.4
500
(continued)
32
-------
TABLE 9 (continued)
Gal.
Gal.
Gal.
Cal.
BOD,
BOD,
COD,
COD,
IDS,
IDS,
TSS,
TSS,
TKN,
TKN,
TP,
TP,
Parameters
TDS, g/1
TKN, mg/1
pH
Turbidity, as ppm Si02
spent brine/ton tanked
spent brine/bu. tanked
process water /ton tanked
process water/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
g/ton tanked
g/bu. tanked
NF
31
328
2.8
400
156
3.9
97
2.4
6,200
155
13,600
340
66,000
1,650
860
21.5
413
10.3
82
2.0
CCF
84
290
2.9
1,000
136
3.4
96
2.4
8,000
200
15,200
380
80,000
2,000
448
11.2
364
9.1
84
2.1
CCF
50
280
3.
625
135
3.
102
2.
6,640
166
8,800
220
48,000
1,200
355
8.
337
8.
79
2.
0
4
6
9
4
0
33
-------
TABLE 10 . SUMMARY COMPARISON OF QUANTITY AND COMPOSITION
OF BRINES FROM NATURAL AND CONTROLLED FERMENTATIONS
Parameters
No . tanks
Spent brine, gal/ ton
Process water, gal/ton
Total water, gal /ton
BOD, g/ton
BOD:N:P
COD, g/ton
TDS, g/ton
TSS, g/ton
TSS, Ib/ton
TKN, g/ton
TP, g/ton
BOD, Ib/ton
Salt, 103 g/ton
NF
4
156
123
279
7,110
100:6:1
13,320*
91,500
665
1.46
444
81
15.6
105
CCF
5
141
125
267
6,730
100:6:1
12,110*
65,120
480
1.06
414
75
14.7
66
*CODs were not available for experiment 2. Value represents average of 2 NF
tanks and 3 CCF tanks.
34
-------
0
So
-------
bloating of cucumbers and that even with a CCF procedure substantial concen-
trations of CCL build up unless sparging is practiced. They noted visible
bloater damage in No. 3 cucumbers when the CO^ concentration was >_ 60 mg/100
ml brine. During the course of our experiments, CC^ was monitored in both
the NF and CCF tanks (Figures 6, 7, and 8). As a comparison, C0£ levels in
tanks in the PPP tankyard were also monitored (Table 11). Note that in all
the experimental NF tanks, the CCL concentration exceeded 60 mg/100 ml during
the active fermentation period, despite the relatively large surface-to-
volume ratio in these tanks. In the 4 large commercial brinings, the CCU
concentrations were excessive throughout the entire first six days in
which they were monitored. In contrast, because of the nitrogen-sparging,
the CCF tanks maintained low C02 levels (<~ 32 mg/100 ml). Obviously,
further studies must be made of the relative contributions to brinestock
quality of culture addition and nitrogen sparging, since the experimental
design of this project does not permit this distinction.
36
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Figure 8. Carbon dioxide accumulation in experimental tanks,
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BRINE RECYCLING STUDIES
Laboratory studies of brine recycling were initially directed toward
finding a coagulant or coagulant aid which, at the low pH typical of spent
brine, would effectively reduce enzyme activity. In addition, an attempt was
made to correlate turbidity removal with reduction of enzyme activity. Pre-
liminary experiments, however, indicated that substantial removals of turbid-
ity could occur with little or no effect on enzyme activity. Since in each
of these cases enough turbidity remained to be visible to the naked eye,
further work was directed to provide a "completely clarified brine," de-
fined as a treated brine which in a test tube does not contain visible
colloids and has a turbidity of less than 7 JTU.
Low pH Coagulation with Sodium Aluminate
Sodium aluminate (NaAlCv,), a coagulant not previously investigated with
spent brines, was evaluated at low pH. It was selected primarily for its
alkaline properties. Characteristic spent brine pH of about 3-2 is below
the range of the isoelectric point for enzymes (Tenney and Stumm, 1965) and
it was postulated that addition of sodium aluminate might raise the pH to a
point at which coagulation of the enzyme and other colloids would occur by
adsorption and charge neutralization. It was also hypothesized that raising
the pH might bring about formation of a "sweep floe" of gelatinous aluminum
hydroxide which would adsorb the enzyme.
Results of the first jar test (Table 12) indicated that increasing doses
of sodium aluminate reduced turbidity of the brine, but even at large doses
residual brine turbidity was still substantial. Although the reduction in
turbidity did not result in a reduction in enzyme activity, these results
were considered inconclusive since the enzyme activities were so high that
accurate determinations were not possible.
Several additional jar tests were performed with sodium aluminate in an
effort to find a reasonable dosage that would provide complete clarification.
Visual inspection, in each case, was sufficient to indicate that even massive
doses (up to 5?6 mg/1 as Al"^) failed to completely clarify the brine.
However, substantial floe formation was observed with sodium aluminate.
Immediately after the aluminate addition, large amounts of floe could be
seen, but the amount appeared to decrease during flocculation and settling.
This behavior could be due to initial formation of aluminum hydroxide on
contact between brine and the coagulant, followed by dissolution of the
hydroxide in the acid environment in the brine; if so, use of sodium aluminate
alone for coagulation would require tremendouse doses to achieve a pH high
enough to maintain the precipitate.
Since these experiments with sodium aluminate did indicate the ability
of aluminum to form a floe in the brine, use of. alum addition and pH adjust-
ment was investigated. Selection of alum appeared more favorable from the
standpoint of coagulant requirements and in addition settling properties of
alum precipitates are frequently superior to those produced by sodium alumi-
nate.
41
-------
TABLE 12. SODIUM ALUMINATE JAR TEST
Sodium
alumina te
dosage (mg/1)
0 (raw brine)
250
500
750
1,000
1,500
Initial
PH
3.2
3.2
3.2
3.2
3.2
3.2
Final
PH
3.2
3.4
3.5
3.6
3.7
3.9
Turbidity
(JTU)
240
190
100
75
56
Enzyme
activity
units
800
800
800
800
800
800
Low pH Coagulation with Alum, AlgCSQ^).1 18
In initial jar tests with alum, the brine was raised with sodium hydrox-
ide to pH 5, a level at which precipitation of aluminum hydroxide would be
expected to occur. The results from the first experiment are shown in Table
13. Although floe formation was observed at all alum dosages, greater amounts
were present at the lower dosages. Some turbidity reduction was realized, but
it was insufficient to actually clarify the brine or to noticably affect
pectinase activity. The turbidity data actually indicated that the lowest
dose (100 mg/1) was more effective than higher doses. In a second jar test
(Table 14) these results were confirmed. In this test pH was monitored more
closely. Addition of alum depressed the pH. Gelatinous floe formed at pH
5.45, which indicated that the brine already contained precipitable metal
species. Turbidity reduction at pH 5.45 was better than at pH 5, but the
brine still had a cloudy appearance.
Increased turbidity removal with incrased pH was further obtained in a
third experiment in which initial pH of the brine was set at pH 7. Results
are shown in Figure 9 and Table 15. Again, significant turbidity removal
was accomplished by simply raising the brine pH, flocculating and settling,
indicating possible presence of aluminum in the brine. To confirm this
hypothesis, metals analyses were conducted, as discussed later below.
Effect of alum dosage on turbidity is apparent in Figure 9 . In-
creasing alum dosage increased turbidity removal to a point at which fur-
ther alum addition resulted in increased turbidity. Addition of alum at
150 mg/1 at pH 7 reduced brine turbidity from 90 to 10 JTU. The treated
brine, though visibly better than that obtained in previous experiments,
still contained faintly visible cloudiness. At the same time, enzyme
activity remained virtually unaffected. Although the enzyme activities
were again so high as to prevent accurate measurement, the data indicate that
42
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43
-------
TABLE 13. TURBIDITY AND ENZYME ACTIVITY AS A FUNCTION OF ALUM DOSAGE,
INITIAL pH = 5
Alum dosage
mg/1*
0
100
200
300
500
700
Turbidity
(JTU)
89
44
48
48
55
54
Enzyme activity
units
385
378
380
400
389
387
*As [A12(S04)3 ' 18H20]
removal of most of the turbidity-causing colloids had little if any effect on
reducing the enzyme activity. Since it was still uncertain, however, if re-
moval of the remaining visible colloids would result in physical removal of
the enzyme, further experiments considered use of coagulant aids to enhance
precipitation,
Low pH Coagulation with Alum and Clay
The first coagulant aid investigated was a clay solution, Nalco 8151,
a slightly anionic bentonite. Use the "optimum" alum dosage previously de-
termined, 150 mg/1, two tests were performed with two different brine samples,
each spiked with pectinase. Results are shown in Tables 16 and 17. Turbid-
ity was not measured in these experiments since in both cases visible colloids
remained after settling and because the major objective was to reduce enzyme
activity. It is apparent from the results that addition of Nalco 8151 at the
selected alum dosage did not reduce enzyme activity. The enayme activities in
each experiment merely indicate expected variations based on the sensitivity
of the enzyme analysis. The results in Table 16 also illustrated the failure
of even large clay dosages to affect enzyme removal. No further investigations
appeared warranted.
Kaolinite was also briefly examined as a possible coagulant aid. Brine
at pH 5.5 with a spiked enzyme activity of about 1?0 units was flocculated with
1000 mg/1 of kaolinite, then settled. Enzyme activity in the supernatant was
l6l, not a significant reduction.
44
-------
TABLE 14 . TURBIDITY AS A FUNCTION OF ALUM DOSAGE,
INITIAL pH = 5.45
Alum dosage
mg/1* Initial pH Final pH
0 (raw brine) 3.2 3.2
0 5.45 5.45
20 5.45 5.4
50 5.45 5.35
100 5.45 5.3
200 5.45 5.2
400 5.45 4.95
*As [A12(S04)3 18H20]
TABLE 15 . TURBIDITY AND ENZYME ACTIVITY AS A FUNCTION
ALUM DOSAGE @ INITIAL j»H = 7
Alum .dbsage Turbidity
mg/1* Initial pH (JTU)
0 3.2 90
0 7 27
50 7 14
100 7 12
150 7 10
200 7 12
300 7 21
500 7 27
Turbidity
(JTU)
90
23
24
25
26
29
34
OF
Enzyme
Activity
units
373
350
577
522
580
570
605
543
*As [A12(S04)3 18H20]
45
-------
TABLE 16. EFFECTS OF NALCO 8151 AND OPTIMUM ALUM DOSAGE,
INITIAL pH = 7
Alum dosage
mg/1*
0
150
150
150
150
150
150
*As [A12(S04)3 '
TABLE
Alum dosage
mg/1*
0
150
150
150
150
150
150
Nalco 8151
mg/1
0
50
75
100
125
150
200
18H2Q]
17. EFFECTS OF NALCO 8151 AND
INITIAL JDH = 7
Nalco 8151
mg/1
0
5
10
20
30
40
50
Enzyme activity
units
185
168
175
180
182
178
140
OPTIMUM ALUM DOSAGE,
Enzyme activity
units
16
21
34
28
35
24
24
*As [A12(S04)3 18H20]
46
-------
TABLE 18. METALS CONTENT IN A NO. 2 SPENT BRINE USED FOR JAR TESTS,
COMPARED WITH THOSE REPORTED BY HENNE AND
Concentration (mg/1)
Metal
Aluminum
Magnesium
Manganese
Zinc
Calcium
Iron
Brine #2
41
58
1.5
1.4
400
10
Geisman brine
13
300
11
12
1,000
40
Metal Analyses of Spent Brines
During earlier jar tests with alum, it was observed that turbidity of the
brine was substantially reduced if the pH was raised to between 5 and 6 and
then the brine was allowed to flocculate (30 min) and settle. Appearance of
a white precipitate at these pH levels suggested that aluminum (or other metal)
might be present in the untreated brine. Subsequent metal analyses revealed
that not only was there a substantial concentration of aluminum, but of other
metals as well. The results of the metal analyses are provided in Table 18
along with those reported by Henne and Geisman (1973), for comparison. The
observed differences would be expected, since metals content of the brines
should vary with differences in soils in which the cucumbers were grown, in
the water used for brine preparation, and in the rock salt used for the brine.
Apparently, some plants also add alum directly to the brines on occasion for
help in controlling unwanted algal growth.
Presence of metals in the brines has several implications. An aluminum
concentration of 40 mg/1 is equivalent to an alum dosage of 4-50 mg/1 as
Alp (SO ). ' HO, a substantial dose in terms of wastewater treatment, yet at
littl
e precipitation was observed. Presence of metals which resist
precipitation during recycling could possibly lead to buildup, which appears
to be the case in studies by McFeeters et al. (1978) though the significance
of the amount they observed may be small.
47
-------
TABLE 19, EFFECTS OF HIGH MOLECULAR WEIGHT, ANIONIC POLYMER
(NALCO 7744A) ON TURBIDITY AND ENZYME REMOVAL AT pH = 5.6
Nalco 7744A
(mg/1)
0
0.1
1
3
10
30
Initial pH
5.6
5.6
5.6
5.6
5.6
5.6
Final pH
5.6
5.6
5.6
5.6
5.6
5.6
Turbidity
(JTU)
24
21
19
20
21
23
Enzyme activity
Knits
135
123
121
134
120
116
Low pH Coagulation, Polymer Addition
Addition of polymers was evaluated in an effort to find some means to
generate the expected precipitation and formation of aluminum hydroxide floe
at pH 5-6. Since the aluminum hydroxide, as well as the enzyme, is expected
to have a slight positive charge at this pH, a high molecular weight anionic
polymer, Nalco ?7^A, was first evaluated. It was hoped that the polymer
might bridge the positively charged particles to form large settleable floe.
Results (Table 19) indicated that a slight improvement in turbidity was ob-
tained with increased polymer dosage to about 1 mg/1. However, all tested
dosages failed to produce heavy floe and brine clarity. There also appeared
to be no significant reduction in enzyme activity. The evaluation of Nalco
7?4^A was discontinued since larger dosages give no indication of improvement
and polymer doses above 30 mg/1 were considered to "be economically prohibitive,
Other polymers were evaluated. Dow A-21, a high molecular weight anionic
polymer, and Dow C-31f a cationic polymer, were tested at pH 5-5 at 0.3, 1, 3>
10, and 30 rng/1, "but neither provided complete clarification or sufficient
reduction of enzyme activity.
Low pH Coagulation with Ludox and Gelite Addition
Since the pectinase enzyme should be positively charged at low pH, it
was hypothesized that the addition of a negatively charged adsorbent to the
untreated brine (pH=3.2) might effect enzyme removal by adsorption. Two such
negative adsorbents were evaluated. Ludox, a negatively charged silica, was
considered a potentially useful adsorbent as its high surface area: weight
ratio provides a large number of adsorption sites per unit dose. Ludox
solution !«iaL6 evaluated in jar tests at doses of 359-3590 mg/1 (Figure 10 ).
As indicated, ^there was no significant reduction of enzyme activity. Further
tests were conducted with Gelite, another negatively charged silica. A brine
48
-------
30
V)
Z
D
>
o
UJ
5
N
Z
UJ
10
1,000
2,000 3,000
LUDOX DOSAGE (mg/l)
4,000
Figure 10. Enzyme activity as a function of Ludox dosage.
49
-------
"spiked" with a larger enzyme dosage was used in these tests, but again no
effects on enzymes activity were seen even at doses as high as 1 g/1 (Fig- 11) ,
Most probable cause for failure of these materials to sorb the enzyme is the
nature of the brine, which contains high salt concentration and large amounts
of organics which could interfere with sorption.
pH Effects on Coagulation and Enzyme Activity
A study of the pH effects on coagulation and enzyme activity was
initiated for two reasons: (l) all attempts to reduce pectinase activity
or generate a completely clarified brine by coagulation and sedimentation at
pH less than 7 were unsuccessful, and (2) the metals analyses indicated that
the brines contained substantial amounts of aluminum, magnesium, and calcium
and it was anticipated that these metals might precipitate at higher pH.
To assess effects of pH, a "spiked" spent brine (designated #2) was
treated at a series of pH values ranging from 3.2 (raw) to 10.1. pH of the
samples was adjusted with NaOH. Following pH adjustment, each sample was
flocculated and settled and the supernatant was withdrawn for enzyme analysis.
At pH 6.7 after flocculation and settling, the brine supernatant was completely
clarified (Table 20). Complete clarification was also achieved at every pH
above 6.7. Of especial significance are the corresponding enzyme activity
measurements. Even though the brine was completely clarified at pH 6.7, no
significant reduction in enzyme activity was observed below pH 10.1, thus
indicating that no correlation exists between enzyme activity and turbidity
removal.
To establish this conclusion, an additional series of samples was
examined, this time adjusting pH with a lime slurry. Effect of pH on
enzyme activity is shown in Figure 12 . Again the brine was completely
clarified at pH 7, but there was no substantial effect on enzyme activity
below pH 9» with the most drastic effects occurring between pH 10.1 and 10.^-.
The fact that significant enzyme reductions did not occur below pH 10.1 even
though massive precipitation and complete clarification occurred at pF as lew
as 6.7 appeared to indicate that reduction of enzyme activity was not the
result of physical removal by adsorption on the precipitate but was, instead,
the result of denaturation at high pH. This denaturation appeared to be
irreversible since readjustment of the sample to pH 5 for enayme analysis
failed to restore enzyme activity to the original value.
Several experiments were performed to confirm these findings. Tests
of spent brine spiked with pectinase enzyme and subjected to high pH by
various procedures indicated that high pH destroyed enzyme activity regardless
of the method of pH adjustment (Figure 13).
Since there was also heavy precipitation and flocculation accompanying
the high pH levels, and since the brines were completely clarified, there
still remained some question as to whether the enzyme was removed physically
by sorption or whether denaturation destroyed enzyme activity. Several
analyses which were performed appeared to favor the denaturation theory.
50
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51
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A sludge sample was removed from a brine at pH 11.5t dissolved, and
the resulting solution adjusted to pH 5- An enzyme analysis indicated an
enzyme activity of about 21 units, about the same as that of the supernatant.
To determine the enzyme activity as a function of pH without interference
from precipitation and floe formation, "synthetic" brines, free of metals,
were produced in the laboratory, one of 50 S acidified to pH 3.2 with acetic
acid; the other, a 35' S 0.1 M lactic acid solution. After "spiking" the
synthetic brines with enzyme, several aliquots were taken from each for pH
adjustment and subsequent enzyme analysis. Enzyme activities of these
brines as a function of pH are shown in Figures 14 and 15 . In both brines
pH decreased sharply above pH 10. However, in the higher salinity brine
difficulties were encountered in maintaining the high pH levels. After allow-
ing the samples to stand for 30 min, pH was again recorded, indicating that the
pH 11.5 brine had dropped to pH 8-9 and possibly accounting for the lesser
reduction expected. In either case, dependence of enzyme activity on pH is
clearly demonstrated. In these brines there was no possibility of physical
removal of the enzyme since there was no precipitation in the solution; the
enzyme activity losses in these brines corresponds to those observed in the
actual brines.
Further confirmation is provided in Figure 16 in which enzyme activity of
a pectinase-containing spent brine from an actual fermentation is plotted as
a function of pH. As in the cases with "spiked" spent brines and synthetic
brines, the enzyme activity decreased rapidly between pH 9 and 11.
Discussion of Failure of Coagulation and Precipitation to Remove Pectinase
In the course of the studies described above, it was observed that in
spite of massive precipitation of aluminum hydroxide and subsequent brine
clarification, enzyme activity remained unaffected, indicating that aluminum
floe had failed to adsorb the pectinase. Several factors described in the
literature may account, at least in part, for this phenomenon. These include
interference by high salt concentration and by competing metal species, as
well as interference by organics (Dixon and Webb, 1964). It is not known
which of these factors, or combination thereof, were responsible for the
observed failure of aluminum precipitation. However, a further experiment
did demonstrate that in itself aluminum hydroxide precipitate is capable of
sorbing the enzyme. A solution of distilled water "spiked" with pectinase
was treated with alum doses of 300 and 500 mg/1. After buffering at pH 5.8
with sodium bicarbonate, the sample was flocculated and settled. Analysis of
the supernatant (Figure 17 ) indicated that enzyme activity was reduced from
220 units to 24 units by the 300 mg/1 dose and further reduced to 15 units by
the 500 mg/1 dose. Since the pH of the samples never was extreme enough to
inhibit enzyme activity these results show that in the absence of interfer-
ences alumina floe is an effective sorbent of the pectinase enzyme.
In summary, removal of pectinase activity by high pH coagulation-
precipitation is due to denaturation of the enzyme at the high pH rather
than to physical removal by precipitation when real or synthetic brines are
employed. Failure of widely used coagulants to remove pectinase activity from
spent brine is probably due, at least in part, to interferences such as salt.
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Due to constraints of space, time, and money, only limited studies were
conducted on recycling spent brines. These laboratory studies were conducted
as described above (p. 18), using large (no. 3) cucumbers and the conventional
NF procedure. Two control pails received fresh 30 *S brine according to plant
specifications. Two pails were brined with recycled NF brine from the tankyard
studies, diluted to 30°S. One of these received brine which had received the
high pH (lime, sodium hydroxide) treatment; the other, untreated brine from the
same tank. The NF brine came from Tank 6, which had a high residual pectinase
activity. The final two pails received undiluted, untreated CCF brine from Tank
5. Salometer data for the studies is shown in the Appendix (Table A-ll). Re-
sults of the studies are shown in Tables 21 and 22, Following active fermen-
tation and a three month curing period, the brinestock was examined for its
quality. While these studies were done on a very limited scale, some ob-
servations seem warranted. Acidity in the pails receiving recycled GCF brine
was high to begin with and increased rapidly in the early part of the fer-
mentation (Table 22). Acidity developed the slowest in the untreated recycled
NF brine and reached the lowest final value(equal, in fact, to the starting
value of the GGF brines). Treated recycled NF brine started with low acidity,
but reached a level nearly as high as that achieved in recycled GGF brine.
The most intriguing results were noted in evaluating brinestock quality
(Table 21). In terms of bloaters, brinestock quality was poorest in the case
of the directly recycled GGF brines. It was conjectured that this was due
to the high initial acidity of the brines and possible inhibition of the desired
fermenting organisms. However, since the pails were relatively shallow they
were not purged. Therefore, an alternate possibility was that in the GGF
brine there was such a high inoculum of the desired organisms that the fer-
mentation was intense and accompanied by unusually extensive carbon dioxide
production, concommitant with bloating. In terms of bloaters, the high quality
of the NF brinestock is worth noting. On the whole, differences among the tests
in terms of firmness (pressure test) is unremarkable. It is remarkable that
untreated recycled NF brine produced firm brinestock, despite the pectinase
content. From a subjective standpoint, those evaluating the brinestock felt
that the quality of brinestock from CCF recycling was substantially poorer from
that from other treatments.
Degree of cure in different treatments deserves comment, as this study
corroborates studies with tankyard recycling of spent NF brines. That is,
degree of cure achieved was more variable and tended to be lower in brinestock
brined in high pH treated NF brine.
ECONOMIC EVALUATION
A realistic evaluation of the differences in cost of the NF and GCF
procedures could not be made. It became apparent early in the study that on
the typical commercial tankyard the monitoring and care of NF tanks was con-
siderably less extensive than that outlined in the recommended procedure and
therefore less extensive than that provided for our experimental NF tanks.
It was our distinct impression that the overall high quality of the NF brine-
stock produced in our studies was due in large part to the care given the
tanks during the early fermentation period. It should be noted, however,
60
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that we did achieve better consistency in the GGF than in the NF fermentations,
despite our care, leading us to believe that while the NF stock can be markedly
improved by closer attention to recommended procedures, it is unlikely to reach
the levels attained in GGF.
In essence, the only extra labor required for GCF is as follows: l)
initial washing of the cucumbers, 2) sanitizing the brine, 3) acidifying
and adjusting the brine, ^4-) adding the culture, 5) monitoring the purging.
If purging is widely adopted for NF brinings, as appears likely, item 5 will
no longer constitute a difference. GGF tanks consistently showed less scum
and film growth than did NF tanks, and consequently required less maintenance
after fermentation was underway.
The only additional capital costs associated with GGF brining are those
for the.purging apparatus and for storage facilities for the starter cultures.
As noted above, the cost differential due to purging will be eliminated if
purging is generally accepted for all brinings. Proper storage of currently
available commercial cultures turned out to be a major nuisance in this project,
since the frozen cultures had to be shipped in dry ice by air freight, picked
up immediately, transported 50 miles to the plant site, and stored at approxi-
mately -70 G until use. Maintaining such low temperatures requires a source
of dry ice and preferably an ultracold freezer. These amenities are rarely
available at commercial tankyards or even in nearby towns. A local dairy
was able to assist us during the project. However, on a tankyard scale,
procuring and storing starter cultures would be a major operation. The
manufacturer of one of the starter cultures (Chr. Hansen) indicates to us
the possibility that there will soon be available commercial freeze-dried
cultures which can be stored in an ordinary deep-freezer.
Water use is initially greater with CGF, since the cucumbers must be
washed before brining. However, less water is required for desalting GGF
brinestock, providing that plant personnel are educated to gauge volume of
desalting water to the amount of salt to be removed from the brinestock.
Relative costs of recycling GGF and NF brines must await further studies.
As indicated above, it appears at least possible, if not likely, that direct
recycling of GCF brines without treatment may not be an option. Obviously,
additional assessment of CGF brine recycle must be made. The aspects briefly
noted above (purging vs no purging; neutralized vs untreated; diluted vs
direct recycle) could be readily studied in the laboratory to determine the
conditions necessary for recycling. Such questions will need resolution before
large scale studies should be attempted with recycled GGF brines, since the
concept is evidently not quite as simple and straightforward as initially
expected.
The major advantage of CGF appeared to be the reliable production of a
consistently high quality brinestock and final product. Less variation was
noted in quality and in course of active fermentation in the case of the
CCF tanks. Taste of the GGF products was quite similar from one brining to
another, in contrast to marked taste differences among NF products. However,
it must be noted that some people do not favor the pronounced lactic acid
flavor of GCF products. With rising costs of raw products (1979 NC costs per
hundredweight: 1's, $12; 2's, $6; 3's, $3.75) it appears likely that
62
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cost due to GGF will be minimal ^hen "balanced against the possibility of losing
a tank of brinestock.
In addition, even if recycling is not practiced, the GGF procedure
greatly reduces the salt content of the wastewaters, an important factor in
producing a satisfactory plant effluent.
In reference to costs of recycling NF brines, Wendle (197?) performed a
desktop evaluation of costs of the three alternate treatment methods usually
suggested in the literature: ultrafiltration, coagulation/sedimentation, and
heat treatment. For the purpose of this cost analysis, all assumptions in
regard to the volume of brine to be recycled were based on operations of the
Perfect Packed Products Go. in Henderson. The size of this plant was assumed
to be typical of the industry. At this facility, during the first six weeks of
green season, there are generated approximately 320,000 gal of 36-hr brines from
brining the smaller cucumbers likely to be associated with high pectinase
activity. This volume would be generated from brining about 60,000 bushels
of no. 1 cucumbers. In addition, about 270,000 bushels of larger sizes would
be brined during the season, producing bout 1,^50,00 gal of spent brine. For
purposes of this comparison, it is assumed that enzyme levels would be
checked prior to treatment, and only those with significant pectinase activity
would be treated. This is estimated to be about 393?000 gal (all from the
no. 1 cucumbers and 5 % from the larger cucumbers).
It was assumed that coagulation/sedimentation would be accomplished by
lime addition to a pH of 11, followed by mixing with compressed air, then
by sedimentation and clarification under quiescent conditions. It was
assumed that ultrafiltration would be accomplished by passage through a
commercial hollow-fiber type unit, at a rate of 10 gpm or 15 gpm, at pore
sizes suitable for pectinase removal (Little et al., 1976). It was assumed
that heat treatment would be accomplished by flash pasteurizagion, by raising
the brine temperature to l65°F for 15 sec, followed by cooling to 50 °F. Costs
of three fuels were estimated, i.e., natural gas, no.2 fuel oil, and propane.
Complete details a.nd calculations are given in Wendle (1977). In brief,
for heat treatment fuel costs alone would be, per year, 3xUK)0 for natural gas,
$2100 for no. 2 fuel oil, and $3000 for propane. Current prices would be
somewhat higher, as costs of fuels are generally rising. For coagulation/
sedimentation, followed by recycle after neutralization with acetic acid,
total annual costs were estimated to be $1900; without the neutralization, $1200.
Costs of ultrafiltration would be largely associated with the capital cost and
the life of the unit would be critical. At 15 gpm capacity with an estimated
10 yr economic life, cost would be per year $2450; with a 20 yr life, $1950.
At 10 gpm capacity, at 10 yr cost would be $2050; 20 yr life, $1650. Labor
costs are not considered for any of these alternatives. Wendle pointed out
that ultrafiltration might provide an attractive treatment alternative since
in addition to economic feasibility there would be the following advantages: l)
the chemical addition needed for coagulation/precipitation would not be a
factor, 2) the brine would be basically unchanged in soluble chemical content,
but removal of enaymes and bacteria would leave a clear sterile brine, 3) wastes
from the process would be minimal, 4) the process would be relatively independ-
ent of rising fuel and chemical costs. The major disadvantage would be that
estimating the life of the equipment under the low pH-high salt conditions
imposed by the contact with the brines is presently impossible. It
63
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would be possible to get realistic information on performance only by large
scale studies over a period of time.
Wendle (197?) also made the interesting observation that each of the
three alternate methods would provide a net savings to the industry just in
terms of reduction of salt use. He noted that under the assumptions he
employed, approximately 120 tons of salt would be wasted by discarding only the
36-hr brines. If only QQ% of this could be recovered for reuse, at a cost of
$32 per ton for rock salt an annual savings of around $3100 could be realized,
more than enough to pay for high pH treatment or ultrafiltration.
64
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REFERENCES
APHA, AWWA, WPGF. 1976. Standard Methods for the Examination of Water and
Wastewater, 14th ed. American Public Health Association, Washington, DC.
*
Bell, T.A., J.L. Etchells, and I.D. Jones. 1955. A Method for Testing
Cucumber Salt-Stock Brine for Softening Activity. Agricultural Research
Service ARS-72-5, U.S. Dept. of Agriculture.
Dixon, M., and E.G. Webb. 1964. Enzymes. Academic Press, Inc., N.Y.
EPA. 197^. Methods for Chemical Analysis of Water and Wastes. US-EPA,
Methods Development and Quality Assurance Research Laboratory, Cincinnati,
OH.
Etchells, J.L., T.A. Bell, H.P. Fleming, R.E. Kelling, and R.L. Thompson.
1973. Suggested Procedure for the Controlled Fermentation of Commercially
Brined Pickling CucumbersThe Use of Starter Cultures and Reduction of
Carbon Dioxide Accumulation. Pickle Pak Sci. 3: 4-14.
Etchells, J.L., T.A. Bell, H.P. Fleming, R.E. Kelling, and R.L. Thonpson.
1974. Bloater Chart. Published and distributed by Pickle Packers
International, Inc., St. Charles, IL.
Fleming, H.P., R.L. Thompson, J.L. Etchells, R.E. Kelling, and T.A. Bell.
1973&. Bloater Formation in Brined Cucumbers Fermented by Lactobacillus
plantarum. Jour. Food Sci. 38: 499-503.
Fleming, H.P., R.L. Thompson, J.L. Etchells, R.E. Kelling, and T.A. Bell.
1973b. Carbon Dioxide Formation in the Fermentation of Brined Cucumbers.
Jour. Food Sci. 38: 504-506.
Fleming, H.P., R.L. Thompson, and T.A. Bell. 1974. Quick Method for Est-
imating GOp in Cucumber Brines. Advisory Statement from the U.S. Food
Fermentation Laboratory, Agricultural Research Service, USDA, Raleigh, NC.
Fleming, H.P., R.L. Thompson, T.A. Bell, and R.J. Monroe. 1977. Effect of
Brine Depth on Physical Properties of Brine-Stock Cucumbers. Jour. Food
Sci. 42: 1464-1470.
Geisman, J.R., and R.E. Henne. 1973. Recycling Brine from Pickling. Ohio
Report 58(4): 76-77.
Henne, R.E., and J.R. Geisman. 1973. Recycling Spent Cucumber Brines.
Presented at 1973 Food Processing Waste Management Conf., Syracuse, NY,
Mar. 26-28, 1973.
65
-------
Little, L.W., J.G. Lamb III, and L.F. Horney. 19?6. Characterization and
Treatment of Brine Wastewaters from the Cucumber Pickle Industry. Report
No. 99, Univ. of NC Water Resources Research Institute, Raleigh, NC.
McFeeters, R.F., W. Coon, M.P. Palnitkar, M. Velting, and N. Fehringer. 19?8.
Reuse of Fermentation Brines in the Cucumber Pickling Industry. EPA-600/
2-78-207, US Environmental Protection Agency, Cincinnati, OH.
Palnitkar, M.P., and R.F. McFeeters. 197^. Use of Recycled Brines for Cucum-
ber Fermentations. Paper presented at annual meeting of Pickle Packers
International, Inc., Chicago, IL, December,
Popper, K., W.M. Camirand, G.G. Walters, R.J. Bouthilet, and F.P. Boyle.
1967. Recycles Process Brine, Prevents Pollution. Food Engng. 39: 78-
80.
Tenney, M.W., and W. Stumm. 1965. Chemical Flocculation of Microorganisms in
Biological Waste Treatment. Jour. Water Poll. Control Fed. 37: 1370-1388.
Wendle, J.G. 1977- Treatment Methods for Removal of Pectinase Activity in
Spent Cucumber Pickling Brines. Master's Report for M.S. in Environmental
Engineering in the Dept. of Environmental Sciences and Engineering, Uni-
versity of North Carolina at Chapel Hill.
66
-------
SECTION 7
LIST OF PUBLICATIONS
To date, publications resulting from this project are as follows:
Little, L. W., R. Harrison, J. Davis, J. Harris, and S. J. Dunn. 1977.
Reduction of Wastes from Cucumber Pickle Processing by Use of the
Controlled Culture Fermentation Process, pp. 322-332, in Proc.
Eighth National Symp. on Food Processing Wastes, EPA - 600/
2-17-184.
Wendle, J. G. 1977. Treatment Methods for Removal of Pectinase
Activity in Spent Cucumber Pickling Brines. Master's report for
M.S. in Environmental Engineering in the Department of Environ-
mental Sciences & Engineering, University of North Carolina at
Chapel Hill. 121 pp.
67
-------
Appendix
TABLE A-l. BRINE CHARACTERISTICS DURING NATURAL FERMENTATION PROCESS
TANK NO. 1, EXPERIMENT NO. 1
Time
Date
June 23
June 24
June 25
June 26
June 27
June 29
July 1
July 5
July 9
July 13
July 19
July 23
Aug. 5
Aug. 11
Aug. 16
Days
0
1
2
3
4
6
8
12
16
20
26
30
43
49
54
Degrees Salom
Top
17
20
20
24
24
24
15
24
30
33
32
23
39
34
Bottom
18
21
22
26
25
25
26
27
34
36
35
40
41
43
% Acid
as
Lactic
0.05
0.21
0.23
0.28
0.38
0.50
0.65
0.68
0.53
0.41
0.48
0.47
0.50
0.45
PH
5.3
5.2
4.6
3.9
3.7
3.4
3.3
3.2
3.2
3.2
3.0
3.2
3.2
3.5
TemD
V
82
87
85
84
87
83
86
82
84
84
85
78
80
85
68
-------
§ »
2 Cft
m o
a> co
IO
(0
in
in
o
in
UlUJ<
QtKtO
in
(0
a;
2
UJ
i
O.
I
i
o
V)
Q
<
I
I
*
Q O
O I-
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in
in
o
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in
in
to
in
CM
in
o
in
CO
o
CD
in
0
in
in
q
in
ro
O
IO
m
CM
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CD
in
in
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to
in
<*
in
ro
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to
in
CVJ
00
CD
CO
Q
UI
z
o
- 2
2 I
u.
co
CO
CM
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c
c
to
co
co
QJ
o
O
a,
c
o
H
4-J
CO
4-J
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0)
B
CO
C
CO
CD
D
O
o
H 4-1
u c
to cu
^ e
n -H
OJ ^J
E OJ
^ ex
CD X
PH OJ
I
<
OJ
>-,
p
M
69
-------
TABLE A-2. BRINE CHARACTERISTICS DURING
CONTROLLED CULTURE FERMENTATION PROCESS, TANK NO. 3,
EXPERIMENT NO. 1
TI'TTI<=>
Date
June 23
June 24
June 25
June 26
June 27
June 29
July 1
July 5
July 9
July 13
July 19
July 23
Aug. 5
Aug. 11
Aug. 16
Days
0
1
2
3
4
6
8
12
16
20
26
30
43
49
54
T><=>gTPp« fSatnm
Top
26
22
21
23
22
25
25
25
25
27
27
27
27
25
Bottom
27
21
21
23
23
26
26
26
27
28
28
27
29
32
"/> Aeid
as
Lactic
0.21
0.27
0.48
0.68
0.86
0.91
0.94
0.97
0.98
1.04
0.96
1.03
1.04
0.84
PH
3.6
4.7
4.0
3.9
3.8
3.6
3.6
3.4
3.3
3.3
3.3
3.5
3.5
3.7
Temp
op
80
86
86-
88
87
83
84
82
80
84
85
77
83
83
70
-------
u.
o
9 in
2 en
Q
o>
10 o
co co
in
o
r-
m
co
o
co
m
io
O
10
Ouj^j
UJUJ<
Q a: co
co in
m
ro
Q
no
in o .«,
oj c\j !2
CL
2
UJ
l-
I
4
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h-
co<
x
a.
m c
5. 10
CO
o
CO
in
10
0
in
o
in
ro
q tf
ro CN
in
in
o
in
in
o
m
ro
o
to
in
OJ
co
CM
CO
OJ ±
UJ
-------
TABLE A-3. BRINE CHARACTERISTICS DURING NATURAL FERMENTATION PROCESS,
TANK NO. 7, EXPERIMENT NO. 2
Time
Date
July 8
July 9
July 10
July 11
July 12
July 14
July 16
July 19
July 21
July 23
Aug. 5
Aug . 11
Aug. 16
Aug. 25
Days
0
1
2
3
4
6
8
11
13
15
28
34
39
48
Deerees Salom
Top
19
24
23
24
24
25
26
25
20
32
35
38
Bottom
22
28
28
28
27
31
30
28
30
33
39
40
% Acid
As
Lactic
0.06
0.08
0.24
0.51
0.63
0.41
0.72
0.77
0.81
0.83
0.71
0.70
PK
4.9
4.0
3.4
3.0
3.0
3.2
3.2
3.1
3.2
3.3
3,4
3.2
Temp
°t
81
86
86
84
85
85
86
85
78
82
80
82
72
-------
8
»
in
oo
o
oo
in
o
r-
m
(O
o
CO
in
in
o
in
08'
UJUJ
UJ<
crco
in
(0
o
co
in
m
o
m
in
M-
a:
2
UJ
X
Q.
o
<
i
<
i
9 u
o H
< o
in
ro
in
CM
q
cvj
t\j 9
x
O.
in
in q
cb co
in
in
0
in
in
in q
ro ro
in
in
o
in
in
o
in
ro
o
JO
in
OJ
00
CO
2
00
CO
^t
CJ
co
UJ
o
c
c
cti
0)
o
o
o
H
C
-------
TABLE A-4. BRINE CHARACTERISTICS DURING
CONTROLLED CULTURE FERMENTATION PROCESS, TANK NO. 8,
EXPERIMENT NO. 2
Time
Date
July 9
July 10
July 11
July 12
July 13
July 15
July 17
July 19
July 21
July 23
Aug. 5
Aug. 11
Aug. 16
Aug. 25
Days
0
1
2
3
4
6
8
10
12
14
27
33
38
47
Degrees Salom
Top
25
23
24
22
23
24
24
24
25
21
27
Bottom
26
24
25
25
25
26
26
25
25
31
28
% Acid
As
Lactic
0.25
0.35
0.47
0.72
0.90
0.96
0.96
1.02
1.12
1.13
0.92
1.03
PH
4.6
4.1
3.8
3.4
3.5
3.4
3.6
3.2
3.6
3.6
3.3
Temp
83
82
82
86
86
86
86
86
74
84
82
74
-------
810 o to o 10 o
_ o> en co co r- r>-
10 o 10 o
tD CO IO 1O
UJUJ<
QtEw
JO O 10 O "O
^ CO 10 tf) ^t
IO
ro
IO
CVJ CM ii
IO
a:
UJ
X
a.
j
4
3
a
u
a o
o r-
<
a *
10
IO
o
IO
10
o
IO
10
o
ro
10
CM
a>
(O
8 ?
UJ
o
* -
~ H-
-------
TABLE A-5. BRINE CHARACTERISTICS DURING NATURAL FERMENTATION PROCESS
TANK NO. 9, EXPERIMENT NO. 2
.Time
Date
July 8
July 9
July 10
July 11
July 12
July 14
July 16
July 19
July 21
Aug. 5
Aug. 11
Aug. 16
Aug. 25
Days
0
1
2
3
4
6
8
11
13
28
34
39
48
Degrees Salom
Top
18
24
23
23
23
25
24
20
32
36
36
Bo 1 1 DIP
20
27
26
25
26
30
29
32
32
39
38
% Acid
As
Lactic
.06
.14
.25
.47
.54
.47
.68
.58
.70
,63
.67
PH
4.9
3.8
3.5
3.2
3.2
3.2
3.0
3.0
3.3
3.6
3.3
Temp
°F
80
84
84
83
83
84
85
77
82
82
82
76
-------
o
o
10
Q
55
"O O
co co
10
10 O
to M
s-i a)
3 a
o x
QJ
o -
H ON
4J
Cfl
4-1 O
C C
0)
G ^i
a) nj
J~T^ 4_J
cu
Sn
3
60
H
I
a.
ID (O 10
10 10 OJ
77
-------
TABLE A-6. BRINE CHARACTERISTICS DURING
CONTROLLED CULTURE FERMENTATION PROCESS, TANK NO. 10,
EXPERIMENT NO. 2
Ti'mp
Date
July 9
July 10
July 11
July 12
July 13
July 15
July 17
July 19
July 21
July 23
Aug. 5
Aug. 11
Aug . 16
Aug. 25
Days
0
1
2
3
4
6
8
10
12
14
27
33
38
47
r)£*crT£ic»G £,a1 rvm
Top
26
24
24
23
24
24
24
24
26
22
26
Bottom
28
25
26
24
26
27
26
26
27
26
27
1
% Acid
As
Lactic
0.23
0.32
0.48
0.70
0,82
0.91
0.97
1.03
1.12
1.07
0.95
0.95
pH
3.6
4.8
4.6
4.2
3.9
3.6
3.4
3.3
3.5
3,3
3.4
3.7
3.4
Temp
°F
82
83
83
84
87
85
87
87
75
85
83
78
-------
2
10
o
m o
oo oo
10
10
CO
10
1O
O
in
cisg
UJUJ<
Qtrco
i
o
co
10
m
o
10
in
10
to
Q
ro
10
CM
8 2
OL.
Z
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g
cj
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4
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to
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to
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to
in
CM
-8s
CO
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CNJ
in
o
in
CD
O
CD
in
in
o
in
in
q
m
to
o
K)
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00
CM
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O
z
UJ
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a:
UJ
en
0)
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o
o
H
4J
cfl
4J
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0)
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a)
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t-i
3
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S-J QJ
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a ai
o o
H iI
4-1
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4-J O
C C
(1)
0 ^
S-i C
a) cd
79
-------
TABLE A-7. BRINE CHARACTERISTICS DURING
CONTROLLED CULTURE FERMENTATION PROCESS, TANK NO. 4,
EXPERIMENT NO. 3
Time
Date
Aug. 3
Aug. 4
Aug. 5
Aug. 6
Aug. 7
Aug. 8
Aug. 11
Aug. 16
Aug. 19
Aug. ,25
Aug. 30
Sept. 2
Sept. 8
Sept. 23
Days
0
1
2
3
4
5
8
13
16
22
27
30
36
51
Degrees Salom
Top
27
21
26
25
24
23
27
28
28
25
25
35
35
Bottom
29
22
27
28
26
24
31
30
30
30
35
36
35
% Acid
As
Lactic
0.27
0.24
0.33
0.68
0.77
1.05
1.05
1.13
1.15
1.40
1.40
1.30
1.30
PH
4.7
4.6
3.9
3.8
3.6
3.6
3.4
3.2
3.2
3.6
Tenro
V
81
81
80
80
77
83
81
78
82
76
69
69
72
80
-------
S 10 o 10 o
2 o> o> to oo
10
o
1^
m o
CO CD
10
to
o
IO
J5
10
o
CD
10
10
air co
o >o
10 ^
IO
ro
o
to
10
evj
IO
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4
i
o
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CSJ _:_:
* 2 CD
10
ip q
co co
IO
o
IO
IO
iq q
to 10
in
csi
o
(O
10
IO
o
10
IO
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to
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to
IO
CM
CO
UJ
00 CD
CO
W UJ
^. UJ
CO
CO
CJ
CO
CD
0)
a
o
a
o
H
4-1
CO
4-1
CU
S-i
0)
M-l
3
O
O
5-i
C
O
O O
C
5-i
O 4-)
CU
co 6
a) -H
> 5-1
5-1 CU
3 a
o K
cu
c
o -
*H ^t~'
4J
rt
4J O
c c
cu
5-i C
CU cfl
J"T| 4J
CU
5-1
3
&0
H
Pn
81
-------
TABLE A-8. BRINE CHARACTERISTICS DURING
CONTROLLED CULTURE FERMENTATION PROCESS, TANK NO. 5,
EXPERIMENT NO. 3
Time
Date
Aug. 3
Aug. 4
Aug. 5
Aug. 6
Aug. 7
Aug. 8
Aug. 11
Aug. 16
Aug. 19
Aug. 25
Aug. 30
Sept. 2
Sept. 8
Sept. 23
Days
0
1
2
3
4
.5
8
13
16
22
27
30
36
51
Degrees Salom
Top
27
21
27
26
25
25
22
26
26
25
25
26
25
Bottom
27
23
28
33
26
26
26
27
27
28
29
27
26
% Acid
As
Lactic
0.26
0.24
0.36
0.56
0.71
0.96
1.03
1.10
1.07
1.31
1.31
1.21
1.17
pH
4.6
4.4
4.0
3.8
3.6
3.5
3.6
3.4
3.3
3.7
Temp
F
81
81
80
80
77
83
86
81
83
76
69
70
72
82
-------
i i
UJUJ<
ace co
CD to 10
o
1O
10
a:
z
ui
i
CL
a u
u
<
T
s
IO
IO
o
10
10
1-
o
IO
CO
o
IO
10
CM
CO
$
a
UJ
oo cp
₯ UJ
u.
00
CD
t
CM
10°
en
en
OJ
y
O
M
a.
a
o
H
4-1
03
0)
4-1
0)
4-101
a
o
a o
a
M
O 4J
IH S
0)
tn e
QJ-rl
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S-i 1)
3 a<
u X
0)
o «
HUI
4-1 O
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-------
TABLE A.-9. BRINE CHARACTERISTICS DURING NATURAL FERMENTATION PROCESS,
TANK NO. 6, EXPERIMENT NO. 3
Time
Date
Aug. 3
Aug. 4
Aug. 5
Aug. 6
Aug. 7
Aug. 8
Aug. 11
Aug . 16
Aug. 19
Aug. 25
Aug . 30
Sept. 2
Sept. 8
Sept. 21
Sept. 23
Days
0
1
2
3
4
5
8
13
16
22
27
30
36
49
51
Degrees Salom
Top
27
20
27
25
26
26
16
28
29
30
27
34
36
39
Bottom
29
23
29
26
27
26
26
30
30
30
30
35
39
% Acid
As
Lactic
0.04
0.05
0.10
0.18
0.24
0.55
0.61
0.76
0.78
0.91
0.89
0.86
0.87
PH
4.8
4.0
3.9
3.8
3.6
3.7
3.4
3.2
3.2
3.5
Temp
F
81
83
78
78
77
82
84
82
80
76
70
70
75
72
84
-------
S to o io o io oioomo
2 Oi (fi 00 00 N- N (0 (0 10 10
UIUI
IO
IO
IO
ro
8
IO
CM
a.
Z
UJ
i
Q.
\
g o
o i-
< u
d
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IO
IO
IO
ro
3
10
CJ
00
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CM u
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U.
co
CO
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c
cfl
CO
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QJ
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C
0)
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CO
0)
S-( CO
3
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c a
o
H 4J
j-i a
Cfl OJ
u 0
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6 a)
M a
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0)
(-1
M
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10
10
« 9
roro
"9
ca
85
-------
TABLE A-10. STATISTICAL ANALYSIS OF ENZYME ANALYSIS DATA
Sample Size
Mean
Variance (s^)
Std. Dev. (s)
95% confidence
Brine #1
(weak)
6.3
4
6
6
5
4
3
4
4
4
10
4.6
1.26
1.12
2.4-6.8
Brine #1
(moderately active)
19
17
20
19
18
16
19
20
8
18.5
2
1.4
15.7-21.3
Brine #3
(extremely strong)
500
350
280
280
300
146
100
250
8
276
15,130
123
30-522
86
-------
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UUJ
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c
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1)
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CC
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o
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H
c
O
Z
CO
CO
[l [
0
o
cc
CL
3
H
O
H
CX
5
O
i-l
CO
O
[5
0
CD
Z
2
CC
CO
i i
O
rH
1
0)
^
5b
H
M
OJ
rH
y
O
H
a
VJ
-------
TABLE A-ll. SALOMETER DATA FOR RECYCLING STUDIES
Fermentation time,
days
0
3
5
7
11
12
18
25
35
Controls
30
21
21
2k
22
24
25
25
31
30
20
18
25
25
24
26
25
31
°S
NF.trt.
30
20
22
24
25
25
26
28
31
NF.untrt.
30
20
24
26
27
30
30
31
31
CCF
25
17
19
22
25
25
28
30
32
25
17
17
22
30
31
31
32
38
88
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/2-80-046
2.
3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
Reducing Wastewater From Cucumber Pickling
Process By Controlled Culture Fermentation
5. REPORT DATE
February 1980 issuing date
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
8. PERFORMING ORGANIZATION REPORT NO.
Linda W. Little, Jeffrey g.
Davis, Robert M. Harrison ,
Wendle, Jeffrey
Samuel J. Dunn
9. PERFORMING ORGANIZATION NAME AND ADDRESS
University of North Carolina at Chapel Hill
10. PROGRAM ELEMENT NO.
1BB610
11. CONTRACT/GRANT NO.
S-804220
12. SPONSORING AGENCY NAME AND ADDRESS
Industrial Environmental Research Lab.-Cin.,OH
Office of Research and Development
U.S. Environmental Protection Agency
Cincinnati, OH 45268
13. TYPE OF REPORT AND PERIOD COVERED
Final Report 1975-79
14. SPONSORING AGENCY CODE
EPA/600/12
15. SUPPLEMENTARY NOTES
North Carolina A&T State University, Greensboro, North Carolina
16. ABSTRACT Qn & demonstration scale, the controlled culture fermentation process
(CCF) developed by the U.S. Food Fermentation Laboratory was compared with the
conventional natural fermentation process (NF) in regard to product quality and
yield and volume and concentration of wastewaters. Weight of cucumbers, volume
of water, and amounts of additives were recorded. pH, acidity, salinity, and
temperature were closely monitored. After brining, brinestock quality was evaluated
by a panel of experts from the US Food Fermentation Laboratory and the Heinz Company.
The brinestock was then processed; spent brines and processing waters were collected.
Volume and wastewater characteristics (salinity, BOD, N and P forms, residues) were
determined for the waters and weight of brinestock was determined. The cucumbers
were then packed using a conventional finishing procedure for whole dill pickles and
hamburger dill chips. Yield of final product was determined. Acceptability of the
finished products was evaluated by a panel.
Analysis of data indicates that the CCF produces a product of quality equal to
or exceeding that of NF; that a reduction of the total dissolved solids load in the
wastewaters was achieved; and that fermentation occurs more rapidly and predictably.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS C. COSATI Field/Group
Food Processing
Fermentation
Quality Control
vvastewater
Cucumber Pickling
Process Modification
18. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY CLASS (ThisReport)
Unclassified __
21. NO. OF PAGES
101
20. SECURITY CLASS (This page)
Unclassified
22. PRICE
EPA Form 2220-1 (Rev. 4-77) PREVIOUS EDI TION is OBSOLETE
ft US GOVERNMENT HUNTING OFFICE 1980 -657-146/559Z
89
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