Water Research Vol. 14, pp.

545 to 551
Pergamon Press Ltd 1980. Printed in Great Britain

THE TREATMENT OF MALT WHISKEY

DISTILLERY WASTE USING T H E FUNGUS
G E O T R I C H U M CANDIDUM
J. P. Q u r ~ and R. MARCHANT
School of Biological and Environmental Studies
New University of Ulster, Coleraine, Northern Ireland

(Received 13 February 1978)

Abstract--The imperfect fungus Geotrichum candidura, when grown in batch and in single and two-stage
continuous laboratory culture, has been shown to produce reductions of up to 92% in the BOD loading
of Irish malt whiskey distillery waste. Maximum COD and TOC reductions were 81% and 76% respect-
ively. Yields of fungal biomass of over 30 g 1-1, containing more than 45% "true" protein, have been
obtained. The economic possibilities of employing such a process for waste treatment and perhaps
single-cell protein production seem encouraging.

INTRODUCTION duction has become increasingly uneconomic due to

the high capital outlay on plant, the necessity to pro-
Current annual production of whiskey in the British
vide biological treatment plant for the highly acidic
Isles totals some 1.9 x l0 s gallons (8.7 x l0 s 1), in the
evaporator condensate, which itself can have a BOD
course of which about 2.1 x 109 gallons (9.6 x 1091)
of as much as 8400 nag 1-1 (Brown et al., 1976) and
of liquid wastes are generated---the equivalent, on a
will be of a similar volume to the spent wash from
BOD basis, of a city of 6.1 x 106 people. The prob-
which it is derived, but particularly because of the
lem of disposing of such vast quantities in the face of
large amount of fuel required to carry out the evapor-
increasingly stringent anti-pollution legislation and at
ation. Shortage of fuel supplies at one Scottish distil-
a time of general economic difficulty has been a very
lery during the "oil crisis" of 1973 led to the dumping
considerable one for distillers (Campbell, 1970; Wil-
of spent wash in disused quarries and caused the local
kin, 19771
water authority to point out the inherent danger to
By far the most important element in whiskey distil-
the quality of groundwaters and local streams (Anon.,
lery wastes, in terms of both volume and organic
1973). It seems inconceivable that energy costs will
loading, is the spent wash which remains after the first
not continue to rise and the economic feasibility of
distillation of alcohol from the fermented wort. It rep-
spent wash evaporation become increasingly ques-
resents approximately 90% of the original fermenta-
tionable.
tion volume. Normally termed "pot ale" in malt whis-
Against this background it seemed important to
key distilleries and "stillage" by grain distillers it is a
investigate the possibility of devising a low-tech-
brown, sweet-smelling, low-pH liquor which is high in
nology microbial treatment process which would not
both dissolved and suspended organic material. Until
only remove the bulk of the organic loading from
early this century it was normally discharged
spent wash and produce an effluent which could be
untreated into rivers, and although this practice has
safely disposed of, but which would possibly refix this
now completely ceased a recent survey of Scottish
organic material as high-protein biomass to serve as a
distilleries by Mackel (1976) showed that 27% were
dietary supplement for animals or even humans. This
still permitted to dispose of their spent wash at sea
concept of upgrading the nutrients contained in an
while a further 10% used public sewage treatment
organic effluent through a microbial digestion process
facilities and 5% sprayed it on to derelict land. Such
is a comparatively recent one, but has akeady been
methods are extremely wasteful and introduce the
tentatively proposed in the context of distillery wastes
possibility of accidental leakage; the alternative tech-
(Tomlinson, 1976; Wilkin, 1977). More intensive
nique, now adopted by about half of the 104 Scottish
studies on other carbohydrate-rich effluent streams
whiskey distilleries and at the two major Irish plants,
have successfully reached pilot plant or even produc-
has been to evaporate spent wash to a syrup contain-
tion level (Jarl, 1969; Birch et al., 1976; Church et al.,
ing 45% solids and at this point to add the spent malt,
1977).
i.e. that fraction which was not solubilized during
extraction of the malt prior to fermentation and nor- METHODS
mally termed "dralI". The mixture is further dried and
pelleted, being sold as "Distillers Dark Grains". a Composition of the waste
low-grade cattle food with a crude protein content of Samples were collected from an Irish malt whiskey distil-
some 23%. In recent years, however, dark grains pro- lery in Bushmills, Co. Antrim, immediately after distillation
545
546 J.P. QUINNand R. MARCHANT
had taken place and were stored deep-frozen until required
for analysis. Prior to such analysis they were filtered
through Whatman No. 1 filter paper, but it was felt that an
adequate determination of suspended solids, reported in
the region of 6g1-1 by Campbell (1970), could not be
adequately carried out on a small sample. Total carbo-
hydrate was determined in the filtrate by the anthrone
method of Fairbairn (1953); total protein was measured
using the biuret method (Herbert et aL, 1971). Free amino
acids were measured by the method of Yemm & Cocking
(1955L and free glycerol by the periodate/acetylacetone
colorimetric technique for triglyceride determination
(Sigma Chemical Company) omitting the saponification
step. Organic acids were detected qualitatively by the
paper chromatographic method of Isherwood and Hanes
quoted in Ranson (1955), and total titrable acidity
determined by titration to pH 7.0 with 0.01 M NaOH
using bromothymol blue as indicator. Total dissolved
phosphorus was determined using the ascorbic acid
method of Herbert et al. (1971), but increasing the concen-
tration of reductant to 0.1 M, and total organic carbon
(TOC) using a Tocsin Aqueous Carbon Analyser (Phase Sep-
arations Ltd., Queensferry, Flintshire). Biochemical Oxy-
gen Demand (BOD) determinations were made according
to APHA Standard Methods 13th edition (1971)--the dilu-
tion water containing seed previously "adapted" to spent
wash--and Chemical Oxygen Demand (COD) determina-
tions by the method of Canelli et al. (1976). Copper was
measured either by atomic absorption spectrophotometry
or colorimetrically after reaction with bathocuproin disul-
phonate (Boehringer Mannheim G.m.b.H). The two copper
determination methods showed good agreement.
Selection of organism
In similar situations, e.g. the work of Smith & Bull
(1976) on coconut processing wastes, the decision was
taken to use a microorganism which is already an
approved food source and therefore invariably one of the
fourteen yeasts listed by Smith (1975). In the present
instance, when the primary aim was to reduce the pollution
load of a waste which is itself the by-product of a yeast
fermentation this approach seemed futile. Enrichment cul-
tures were accordingly set up using inocula from a variety
of natural habitats including aerobic and anaerobic soils,
and activated sludge plants treating domestic and food
processing wastes. In all instances the predominant or-
ganism in such cultures was identified as the imperfect
fungus Geotrichum candidum Link ex Pers.; the strain iso-
lated from a municipal activated sludge plant in Coleraine
was chosen, and maintained as a stock culture on spent
wash solidified with agar (1.3~ w/v).
Batch culture experiments
Batch culture experiments were normally carried out
using 25 ml samples of spent wash in 250 ml Erlenmeyer
flasks, inoculated with 0.2 ml of a suspension of stale G.
candidum arthrospores, which had been precultured in the
same medium. The spent wash was collected as required
(normally monthly) immediately after distillation, and the
supernatant decanted after having been allowed to settle
for a few hours so that its suspended solids content was
negligible. It was then sterilized by autoclaving for 20 min
at 115°C, preliminary experiments having shown that
autoclaving of the growth medium had no effect on the
subsequent yield of organism. Preliminary work had also
indicated that the growth rate of the organism was
retarded if the Erlenmeyer flask contents exceeded 10~o of
the flask volume.
The flasks were incubated at 22°C in a New Brunswick
reciprocal shaker (120 rev rain- 1). Early batch experiments
were carried out using spent wash diluted rivet'old with
distilled water--in later experiments, and in all continuous
culture work, the waste was used undiluted. Dry weight
determinations were carried out 'after filtration of repli-
cated volumes of the culture through dry, pre-weighed
Oxoid 0.45/tin membrane filters. These were then washed
and dried to constant weight at 40°C prior to a final re-
weighing. The culture filtrates were collected and stored
deep-frozen; analyses of pH, BOD, COD and TOC were
later carried out.
To ascertain if supplementation of the spent wash would
be required on a routine basis to obtain maximal reduction
of its organic loading and yield of G. candidura, samples
collected on four separate occasions were supplemented
with NH,NO3, (4gl - t ) KH2PO,, (4g1-1) urea, 14gl - l )
or glucose (10 g l- 1), and inoculated and incubated for 40 h
as described previously.
Similar 40 h batch culture studies were carried out on
twelve samples of unsupplemented spent wash collected on
separate occasions over the period 1976-1977.
Continuous culture experiments
(a) Single stage. These experiments were carried out in a
glass, water-jacketed, column fermenter of 350 ml capacity
(Fig. 1), latterly at a temperature of 22°C when it has been
established that this was optimal. Aeration and mixing
were achieved by the passage of pre-humidified air through
a sintered plate in the bottom of the fermenter, supplied at
the rate of 6 culture vols min- 1 to ensure adequate mixing,
since this is often difficult to achieve in column fermenters.
Because of the relatively small volume of the fermenter the
sample size for biomass determinations and chemical
analyses was kept to a minimum--if possible 25 ml; sam-
piing was carried out when a steady state had been
attained at each dilution rate studied--normally this
required the passage of three to five culture volumes of
medium. The spent wash, which was supplied to the fer-
menter via an LKB Varioperpex peristaltic pump, had
been sterilized after collection to permit its storage; other-
wise no precautions were taken to ensure axenic operation.
(b) Two stage. Two fermenters similar to that described
above were arranged in series, so that the effluent from the
first became the input to the second (there was no attempt
at biomass separation between the stages). Dilution rate in
the first fermenter was maintained at 0.125 h - 1; by altering
the relative culture volumes in the two fermenters (and
their air flows accordingly) dilution rates could be achieved
in the second ranging from 0.085 h - 1 to 0.15 h- 1.
RESULTS
Analytical details of some of the various spent wash
samples collected during the course of this work are
presented in Table 1 ; those relating to a single sample
of a local grain distillery spent wash lcollected
October 1977) are included for comparison. In addi-
tion qualitative paper c h r o m a t o g r a p h y indicated that
only two organic acids were present, lactic and succl-
n i c - - t h e latter in much smaller quantities. Specific
chemical a n d enzymic analyses for these have proved
ineffective to date, presumably because of the general
complexity of the liquor a n d also because of its high
b a c k g r o u n d colour which makes colorimetric
measurement extremely difficult. Accordingly, total
titrable acidity has been expressed as lactate.
Figure 2 illustrates a batch culture experiment in
which the unsupplemented spent wash used as growth
m e d i u m h a d been diluted fivefold with distilled water.
The growth curve shows two growth phases---the first
e x p o n e n t i a l - - a n d a diauxic lag. Cultures grown on
other spent wash samples did not always reveal the
r

Table 1. Analytical details of whiskey spent wash samples

Total
titrable Total
Free acidity dissolved
Date of Carbohydrate Protein amino acids Glycerol (as lactate) BOD COD TOC phosphorus Copper
collection pH gl-I gl-I g l -I gl t gl-t mgl-i mg 1-1 mg I - l P.P.M. P.P.M
?,
1976
May 3.80 13.5 19.8 2.9 5.0 5.8 28,500 47,000 18,820 1270 <0.2
June 3.25 I 1.0 21.5 3.9 6.4 14.0 35,500 42,250 25,620 740 0.4
Sept. 3.20 6.9 15.1 3.4 4.4 11.6 27,500 47,000 35,000 960 2.0
Nov. 3.30 1!.6 17.3 2.8 5. I 9.2 26,000 48,000 N.D. 1280 1.2
Nov. 3.30 9.5 19.0 3.2 5.1 12.3 30,000 52,000 24,320 1140 N.D.
1977
Jan. 3.20 7.0 17.5 2.8 4.4 12.4 27,500 35,500 21,440 1310 1.9
Mar. 3.10 I I.I 23.0 3.3 5.6 17.6 40,500 51,000 30,020 1380 3.5
May 2.90 20.0 24.1 2.1 5.4 18.4 38,500 58,500 28,660 1300 N.D.
June 3.20 8.0 17.3 2.1 6.1 13.5 24,500 35,500 20,660 1180 4.8
Aug. 3.85 21.2 26.5 2.2 6.9 4.9 33,000 52,000 18,820 1460 4.8
Sept. 3.35 6.7 23.0 3.4 6.2 10.7 28,500 36,000 21,440 1570 0.5
Oct. 3.20 10.4 31.0 4.3 7.5 18.4 43,000 47,500 32,780 1210 0.2
Oct.* 3.20 2.9 4.9 0.6 2.6 2.3 10.500 15,000 4920 1960 N.D.

* Grain whiskey spent wash.

N.D. not determined.
548 J.P. QUINNand R. MARCHANT

Hedium

4°r
C0o(ing
30.

~ 2o. 80

60
10-

40
(
00( • -3-0
10 20 3O
Time (hi

Fig. 2. Batch culture of Geotrichum candidum grown on

distillery spent wash (diluted fivcfold). O dry weight, • pH,
TernS,,ture C0rltrol t ~ ~ Spent

Woter ~-~ , 5 cm [;uiture

Fig. 1. Diagram of the basic column fermenter used in
continuous culture experiments.
second growth phase and/or diauxic lag so clearly.
Specific growth rate during the first phase of exponen-
tial growth was 0.250 h - t, and during the initial part
of the second phase 0.217h-t; later experiments
using undiluted spent wash samples showed specific
growth rates for the first exponential phase of as high
nutrient status of the unsupplemented spent wash (as
reflected in the level of biomass production) was rela-
tively low e.g. November 1975, December 1975, such
supplementation appeared to have negligible effect;
when larger amounts of carbon source were available,
however, the addition of P or organic nitrogen
appeared to increase biomass production. The effec-
tiveness of supplementation with inorganic N as
NH,NO3 in such instances is unclear. Since the
fungus, however, was able to convert to biomass ad-
ditional C added as glucose to the otherwise unSUl>
plemented wash with an efficiency of about 50% (ap-
proximately that obtained in studies using defined
media), and in view of the fact that supplementation
did not appear greatly to improve the quality of the
10.0OO'

as 0.385 h - t although this varied widely. A shar p rise

in pH was associated with the end of the first phase of
growth; this phenomenon was observed in all the
spent wash samples in which growth of the fungus
was studied.
Figure 3 illustrates the decline in BOD, COD and
TOC of the culture filtrate in the course of the batch E
experiment. After 34h these showed decreases of
92.2% (BOD), 80.6% (COD) and 63.7% (TOC) on
their initial levels. Productivity of the culture over the
34h before stationary phase was reached was
0.106 g 1 1 h- l, rate of T O C removal 0.08481 - 1 h - l
-

and yield of organism in terms of organic carbon util-

ized 126.4%. (It was felt that such values as yield and
rate of substrate utilization would be best calculated
in terms of total organic carbon, this being the par-
ameter whose determination appeared least subject to
experimental error.) Time (h)
Table 2 illustrates the effect of supplementation of Fig. 3. Batch cultuxe of Geotrichum c ~ grown on
the spent wash growth medium with N H , N O 3 , distillery spent wash: BOD (O), COD (O) and TOC (I-1)
KH2PO,, urea and glucose. When the overall values of culture filtrate.
 Treatment of whiskey distillery waste 549

Table 2. Biomass production by Geotrichum candidum cul- Table 5. Percentage decrease in BOD, COD and TOC
tured in batch on spent wash with various supplementa- achieved as a result of the batch culture of Geotrichum
tions. Levels of supplementation were: NH,NO3 4 g i- t; candidura on samples of unsupplemented spent wash
KH2PO, 4 g l - t ; urea 4 g l - t ; glucose 10g1-1
BOD % COD % TOC
Date of Biomass Date reduction reduction reduction
sample Supplementation production (g 1- t)
1976
Nov. 1975 NIL 17.6 May 64.9 31.7 22.5
NH4NO3 14.8 June 73.2 38.6 58.2
KH2PO4 18.0 Sept. 74.1 61.5 51.0
UREA 18.4 Nov. 63.5 48.4 N.D.
GLUCOSE 22.4 Nov. 68.3 53.3 55.6
Dec. 1975 NIL 17.6 1977
NH,NOa 17.2 Jan. 74.5 50.6 52.5
KH2PO, 17.2 Mar. 91.4 68.0 61.9
UREA 18.4 May 87.0 77.8 75.6
GLUCOSE 21.6 June N.D. 68.9 58.2
March 1976 NIL 20.4 Aug. 72.1 57.4 61.1
NH4NOa 25.6 Sept. 75.7 76.5 N.D.
KH2PO4 23.2 Oct. 70.4 68.6 69.4
UREA 22.0
GLUCOSE 25.6
August 1977 NIL 27.2 such errors are magnified by the very large dilutions
NH,,NOa 26.0
KHzPO4 29.6 of the waste which were necessary for most determin-
UREA 31.2 ations. In this instance the increases in diluent BOD
GLUCOSE 32.8 and TOC compared with the effluent from unsupple-
mented waste are in good agreement.
Accordingly the data reported in Tables 4 and 5 on
final culture filtrates, it was decided that such addi- the reduction of the organic loading of a series of
tions would not be made. The l o w effluent COD spent wash samples, through the batch culture of G.
value of 18,000 after supplementation with urea candidum, and on the biomass yields thus produced
(Table 3) must be attributed to experimental error-- were obtained without supplementation. Maximal
rate of TOC removal obtained in these experiments
Table 3. The effect of supplementation on the quality of was 0.54 g l - ' h - t and of biomass production
the final culture filtrate after batch growth of Geotrichum 0.68 g 1- t h - 1 after an arbitrary 40 h incubation.
candidum in spent wash (collected August 1977) Nor was supplementation practised in the experi-
ments carried out on the continuous culture of the
BOD COD TOC
Supplementation (mg 1-1) (mg !- t) (mg 1- t) fungus on spent wash. In a preliminary experiment in
which a fixed dilution rate of 0.125 h -~ was main-
NIL 6000 23,500 7320 tained and temperature varied between 14.5°C and
NH,NOa 6000 26,000 N.D. 40°C the optimum temperature in terms of TOC
KH2PO4 6000 28,500 7320 removal and biomass production, was found to be
UREA 6500 18,000 8360
UREA + KH2PO4 7000 26,000 8880 - 22°C. This was the temperature chosen for the sub-
sequent experiment in which dilution rate was varied.

Table 4. Biomass production, pH, and residual BOD, COD and TOC levels after
batch culture of Geotrichura candidum on samples of unsupplemented spent wash

Residual Residual Residual

Date of Biomass BOD COD TOC
sample (g 1- t) (mg 1- ') (rag 1-1) (rag 1- ') pH

1976
May 10.6 I0,000 28,000 12,700 6.9
June 21.6 9500 27,000 10,720 6.8
Sept. 16.0 7500 15,000 8580 6.9
Nov. 13.4 9500 25,000 11,380 6.8
Nov. 16.4 9500 25,000 10,810 6.8
1977
Jan. 14.6 7000 20,500 10,180 6.9
Mar. 18.6 3500 20,000 11,440 6.2
May 26.2 5000 14,000 7000 6.9
June . 15.0 10,000 14,500 8640 6.2
Aug. 26.5 6000 23,000 7320 6,8
Sept. 19.5 4500 10,000 N.D. 6.8
Oct. 24.5 8000 19,500 10,040 6.7
550 J. P. QUINt<and R. MARCHANT

Table 6. Analyses from a two stage continuous culture of Geotrichum candtdum on distillery spent wash

o~, Biomass
Dilution increase
rate Biomass in second Filtrate "i, TOC Filtrate % BeD
(h - i) pH (g 1- 1) stage TOC removal BeD removal

First 0.125 3.35 15.6 24,800 24.4 20,000 25.9

stage
Second 0.10 4.70 31.0 98.7 19,720 39.8 13.500 500
stage
First 0.125 3.70 25.6 19,720 39.8 15,000 44.4
stage
Second 0.125 6.60 33.2 29.6 17,180 47.6 10,000 63.0
stage
First 0.125 4.20 28.8 N.D. N.D. 13,000 51.9
stage
Second 0.15 6.60 35.2 22.2 18.500 43.6 4000 85.2
stage
First 0.125 4.80 20.8 17,800 45.7 12,500 53.8
stage
Second 0.085 6.80 34.0 63.5 9900 69,8 3500 87.0
stage

It must be emphasized that during this experiment,
despite the absence of aseptic precautions, contami-
nation did not occur at any dilution rate.
Wash-out took place at a dilution rate between
0.175h -1 and 0.200h -1, while substrate utilization
and biomass production were greatest at the lowest
dilution rate (0.025 h - 1). A dilution rate of 0.125 h - 1
however, proved optimal in terms of biomass produc-
tivity ( 2 . 2 4 g l - t h -1) and rate of TOC removal
( 1 . 5 3 g l - l h - 1 ) ; yield was 1.46g (g C utilized)-1.
However, at D = 0.125 h -~ BeD, COD and TOC
reductions of only 50.0~/~ 30.09/~ and 40.7% respect-
ively were obtained, and it was felt that by the addi-
tion of a second fermenter, coupled in series with the
frst, the high productivities and yield associated with
the higher dilution rates could be retained, whilst also
achieving a high concentration of biomass and a low
orgamc loading in the final effluent--features associ-
ated with the lower dilution rates.
The results produced by the second stage in such a
two-stage system, in which the dilution rate in the
first fermenter was maintained at 0.125 h-1 and that
in the second varied between 0.085 h-1 and 0.15 h - 1
are shown in Table 6 together with the values simul-
taneously attained in the first fermenter, since it
proved impossible to maintain a steady state in the
first fermenter over the entire 300-h duration of the
experiment. (The reasons for this are currently under
investigation and are possibly related to shifts in
population morphology or a result of the sel~tion of
an improved strain of the organism in the highly
selective environment of a high dilution rate con-
tinuous culture regime.)
On the basis of Table 6 it can be seen that with an
overall retention time between the two stages of
19.75 h it was possible to effect an 87% removal of
B e D : this corresponds to a rate of TOC removal of
1.16 g 1- ~ h - I and to a yield of 1.48 g (g C utilized)- i.
Overall productivity was 1.72 g 1-1 h - 1. However,
since this was a non-aseptically operated process it
might be preferable if a dilution rate in the second
stage were chosen such that the pH of the culture did
not rise so close to neutrality, for at D = 0.085 h -
first signs of bacterial contamination were observed.
Dilution rates in the two stages of 0.125 h -1 and
0.10 h-1 would seem to adequately optimize produc-
tivity and substrate utilization respectively.
Biomass removed from the second fermenter when
operated at a dilution rate of 0.125 h -~ was found to
have a "true" protein content (as measured by the
Biuret reaction) of 45.5%.
DISCUSSION
The composition of distillery spent wash has
received little detailed attention in the literature,
partly because of the variability produced by vari-
ations in malt quality or in the conditions of the
manufacturing process, and partly because it has
never been regarded as a particular asset by distillers.
The present data indicate that carbohydrate and
protein form the bulk of the dissolved organic content
of spent wash: the wide variations encountered in
their levels is probably an indication of the varying
degrees to which saccharification of the grain polysac-
charide can proceed during the malting and mashing
processes. It is anticipated that most of the anthrone-
positive material would consist of limit dextrins and
maltotetraose and more complex oligosaccharides,
while Patel & Ingledew (I973) have shown that wort
protein is not used as a carbon source by Saccharo-
myces carlsberoensis during brewery fermentations.
The pH of tbe liquor is consistently low despite the
very considerable buffering effect exerted by its com-
poaents. The lactate fraction of the total titrable
acidity is due to the inevitable contamination of the
fermentation by the rich distillery microfiora. Both
succinate and glycerol, however, are by-products of
 Treatment of whiskey distillery waste
yeast metabolism (Oura, 1977) and are formed in
strictproportion to the amount of ethanol produced.
The copper content of the wash--derived from the
distillery fittings--varies considerably, but does not
appear to bc correlated with its p H as might have
been anticipated.
B O D : P ratios arc low--on average 30:1--~espitc
the extremely high values for B O D and C O D loading
which indicate that malt distilleryspent wash is per-
haps the most concentrated organic waste produced
in the British Isles (cf. Tomlinson, 1976). While tbe
disparity between B O D and C O D values shows that
a significantproportion of the organic content is not
biodegradable, it yet seems one of tbe most appro-
priate substrates with which to conduct a biological
treatment process combining removal of organic
matter with its refixation as high-protein biomass.
The fact that such a process can be carried out by
an organism previously cultivated for human con-
sumption because of its high protein content and
good harvestability(Ledingham et al., 1946) and that
aseptic precautions need not be preserved to maintain
a monoculturc would seem to increase considerably
its economic viability.
Even if it is judged purely as a waste treatment
process it would appear to have advantages over the
evaporation methods currently employed. Effluents of
a strength no greater than "evaporator condensate"
could certainly be achieved by a batch or continuous
two-stage mode of operation, and with the advantage
that they would not require neutralization or nutrient
supplementation before final treatment by conventio-
nal biological filtration. Indeed the provision of bio-
mass feedback would almost certainly make a single-
stage continuous process feasible, and at quite high
dilution rates.
In terms of removal of organics the results of the
present study compare very favourably with those of
other workers who have investigated similar pro-
cesses. Shannon & Stevenson (1975) using eight fungal
and yeast strains achieved BOD removals from brew-
ery effluents of 15.5-56.2% after laboratory batch cul-
ture for up to 8 days, while Updegraff et al. (1973)
obtained a TOC reduction of 50-80% in batch and
48% in continuous culture in coffee wastes using a
variety of fungi. Most efficient removals appear to be
those of Church et al. (1973) who achieved 95% BOD
reduction in corn and pea canning wastes using Tri-
choderma viride and Gliocladium deliquescens in a
non-aseptic continuous process operated at a dilution
rate of 0.05 h = 1 in 10,000 gallon tanks. Initial BOD
values, however, were relatively low, about
1600 mg 1 - 1 . Interestingly, G. candidum was an impor-
tant contaminant in the process, occasionally becom-
ing dominant.
Intensive studies on optimization of the conditions
for biomass production and on the nutritional value
of the product are currently being carried out.
Acknowledgements--This work was supported by a
551
Research Studentship awarded to one of us (J.Q.) by the
Northern Ireland Department of Education. We also grate-
fully acknowledge the co-operation of the staff of Old
Bushmilis Distillery Ltd., and of Mrs. Mary Parr of the
Northern Ireland Department of Agriculture Freshwater
Biological Investigation Unit who carried out the TOC
analyses.
REFERENCES
American Public Health Association 0971) Standard
Methods for the Examination of Water and Wastewater.
American Public Health Association, Inc., New York.
Anon (1973) Annual Report of the Banff, Moray and Nairn
River Purification Board, p. 18. Elgin, Morayshire, U.K.
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