The tolerance and nutritional value of two

microfungal foods in human subjects13

John N Udall,4

Clifford

W Lo,

Vernon

R Young,

and

Nevin

S Scrimshaw

KEY

WORDS

Microfungal

food,

Introduction
With increasing
pressure
on the worlds
agricultural
and fishery
resources
from
an
expanding
population,
the search
for new
and abundant
protein
sources
for human
consumption
has turned
to single-cell
proteins. Studies
of their nutritive
value in experimental
and farm animals
indicate
them
to be a source
of protein,
comparable
to
casein,
and of higher
quality
with added
methionine
(1-3).
In human
feeding
trials,
however,
both bacterial
(4) and yeast proteins (5) have often
been
associated
with
adverse
side effects.
Gastrointestinal
symptoms have been observed
when some microbial proteins
were consumed
in amounts
as
low as 10 g/day (6). Rashes
have also been
associated
with the consumption
of microbial proteins
(7). Moreover,
the relatively
high purine
content
of single-cell
organisms
results
in increased
blood and urinary
uric
acid in individuals
ingesting
them in significant amounts
(8). Processing
to reduce
the
RNA content
solves this problem
(3, 9, 10).
There is only one published
report
of the
acceptability
and nutritional
value of a microfungal
food product.
A novel food from
the hyphae
of the mushroom,
Polvporus
The American
Journal
of Clinical
Nutrition
40:
1984 American
Society for Clinical
Nutrition

AUGUST

single-cell

protein,

novel

food,

nutritive

value

squamosus-64,
produced
in submerged
culture in Bulgaria
has been fed to farm animals
and humans
without
adverse
side effects
( 1 1).
The tolerance
and nutritional
value of two
similar
microfungal
proteins
for humans
are
the basis of this report.
Both novel proteins
were extensively
studied
in animals
before
the human
studies
(12-14).
There
were no
signs oftoxicity
or carcinogenicity
associated
with the use of these novel foods in experimental
animals.
Animals
thrived
and gained
weight
when fed diets that contained
up to
40% (w/w)
microfungal
products
(12- 14).
The human
studies
were approved
by the
I

From

the Department

of Nutrition

and

Food

Sd-

ence and the Clinical

Research
Center,
Massachusetts
Institute
of Technology,
Cambridge,
MA 02139.
2 Supported
in part by the National
Institute
of
Health/General
Clinical
Research
Center Grant 5MOlRR00088-20,
by Ranks
Hovis
McDougall
Ltd. High
Wycombe,
England
and by Tampella
Process
Engineering Co, Tampere,
Finland.
3 Address
reprint
requests
to: John N Udall,
MD,
Clinical
Research
Center,
50 Ames Street,
Massachusetts Institute
of Technology,
Cambridge,
MA 02142.
Recipient
of a Clinical
Associate
Physician
Award
(SMO l-RR00088-20).
Received
September
26, 1983.
Accepted
for publication
February
28, 1984.
1984,

pp 285-292.

Printed

in USA

285

Downloaded from www.ajcn.org by on November 11, 2010

ABSTRACT
The tolerance
ofhuman
subjects
to two microfungal
food products
was studied
in separate
double-blind
cross-over
studies.
As an addition
to the subjects
usual diets, cookies
with and without
20 g of a product
from Fusarium
graminearium
were fed to a group of 100
individuals
daily. In a second
study, cupcakes
with and without
10 g of Paeciomyces
variotii
were given daily to 50 individuals.
Mild rashes possibly
related to one of the microfungal
food
products
occurred
in two individuals
fed P variotii.
Except for a decrease
in serum cholesterol
during the F graminearium
study, no significant
changes
were noted in I 7 serum constituents.
During
nutritive
value studies,
digestibility,
biological
value, and net protein
utilization
were
calculated
for the two microfungal
proteins
and for milk. The values for milk were 95, 85, and
80%, respectively.
The values for F graminearium
were 78, 84, and 65%, respectively.
For P
variotii corresponding
figures were 81,67, and 54%. On the basis of these resultsboth microfungal
foods may be deemed
safe for human
consumption
at the levels tested.
Am J Clin Nuir
I 984;40:285-292.

286

UDALL

ET

and methods

The nature and intent of each study was explained

thoroughly
to participants
and a consent
form was
signed by each individual.
A careful
medical
history
was obtained
followed
by a complete
physical
examination.
The tolerance
and nutritive
value studies were
under the supervision
of a physician
who was available
to evaluate
any symptoms
that occurred.
Participants
were asked to keep personal
diaries and to record any
unusual
symptoms.
Data from the studies
were analyzed using both paired and sample I tests (15).
Tolerance

studies

Subjects.
A total of 100 healthy
adult volunteers
participated
in the Fusarium
graminearium
protein
(FGP) study and 50 in the Paecilomyces
variolii protein
(PVP)
study.
The mean
age, weight,
and height
of
subjects
participating
in each study are shown in Tabk
Study design.
Participants
in the studies
remained
on their usual diets and went about
their customary
activities
during both study periods.
Each study lasted
67 days and followed
a double-blind
cross-over
design
(Fig I). For 30 days each, subjects
ingested
either
a
control food or an experimental
microfungal
food. The
foods were eaten daily at the Massachusetts
Institute
of
Technology
research
kitchen
5 days/wk.
Subjects
were
given
a ration
of either the control
or the microfungal
food to take home for Saturday
and Sunday.
Initial
assignment
to the control
or experimental
food period was random.
After the first 30-day period,
the control
or experimental
food was stopped
for 1 wk
before subjects
were changed
over to the alternative
food. The foods were coded in such a way that neither
the physicians
nor the subjects
knew whether
a participant was receiving
the control
or the microfungal
preparation.
Diets. The first study was conducted
with FOP produced
by Ranks
Hovis
McDougall
Ltd. High Wy-

TABLE
1
Characteristics

of subjects

participating

Study

in microfungal

No/sex

feeding

studies

Age

(mean

Wt and

Ht
(rn

Studies

PVP

Nutritive
FGP
PVP

range)

kg

1T

Tolerance
FGP

and

Wt

value

ht of subjects

50/F
50/M
25/F
25/M

23.7(18-59)
22.6 (18-40)
25.6(18-53)
22.2(18-30)

62.0(43-86)
72.0 (56-88)
61.0(45-80)
72.0(51-97)

167 (153-178)
177 (160-195)
160(156-178)
179(160-196)

13/M
6/M

19.9 (18-25)
20.3(18-22)

73.0 (59-86)
66.0(60-75)

180 (169-187)
175 (163-186)

studies

obtained

during

the physical

examination

before

each

study.

Downloaded from www.ajcn.org by on November 11, 2010

combe,
England,
and commercially
available
as Mycoprotein.
FGP was produced
by continuous
fermentation
on a medium
of commercial
glucose
syrup, treated
to
reduce the nucleic acid content
and dried. The product
contains
54. 1% protein
(N x 6.25) by weight (43.7%
true protein),
14.9% fiber, 5.8% moisture,
6.1% ash,
and 1.0% ether extract (12). The amino acid composition of FOP is shown in Table 2. It was fed in experimental
cookies
each containing
S g of FGP (2.7 g of
protein)
or otherwise
identical
cookies
without
the
added
FOP. Subjects
ate two cookies
each morning
between
9:00 and 10:00 AM and two cookies
in the
afternoon
between
4:00 and 5:00 PM. Therefore,
participants
consuming
the experimental
cookies
ingested
20 g ofFGP
per day.
For the second
study, PVP was produced
by Tampella Process EngineeringCompany,
Tampere,
Finland,
on an energy substrate
of the spent liquor of a sulfite
wood pulping
mill (13, 14). The product
is an almost
odorless,
cream-colored
powder
that is 55.9% protein
by weight
(46.2%
true protein),
28.6%
nitrogen-free
extract,
8.8% crude
fiber, 5.6% ash, and 1.1% ether
extract
(13, 14). It is commercially
available
as Pekilo
protein
and was not treated to reduce the nucleic
acid
content.
The amino acid composition
of PVP is shown
in Table 2. A commercial
cake mix was used in the
Massachusetts
Institute
ofTechnology
research
kitchen
to prepare
muffins
with and without
10 g of PVP (5.6
g of protein).
Subjects
ate one muffin a day.
Specimen
collection.
Blood specimens
obtained
during each of the two tolerance
studies
were drawn
between 8:00 and 9:00 AM after an overnight
fast. Four
25.0-ml
venous
blood samples
were taken from each
participant
during
the FGP study.
One sample
was
obtained
before and after each ofthe two study periods
(Fig 1). Three
venous
blood
samples
were obtained
during
the PVP study. One sample
was drawn
before
the initial 30-day study period,
a second
sample
after
this period,
and the third at the termination
of the
second 30-day period (Fig 1).
The concentrations
of 17 serum constituents
were
measured:
glucose
(Sigma Bulletin
635, Sigma Chemical Co. St Louis,
MO), blood
urea nitrogen
(BUN)
(Hyland
UN-Test
Reagent
Kit, Hyland
Division
Trayenol Laboratories,
Inc. Costa Mesa, CA), sodium
and
potassium
(analyzed
on an IL 443 flame photometer,
Instrumentation
Laboratory,
Inc. Lexington,
MA), cal-

Committee
on the Use of Humans
as Experimental
Subjects
and the Executive
and
Policy Committees
of the Clinical
Research
Center
of Massachusetts
Institute
of Technology.
Materials

AL

MICROFUNGAL
TOLERANCE

STUDIES

30

DAY 0

287

FOOD

67

37

CONTROL

CONTROL

DAY

%r%rJ3

11

CONTROL

CONTROL

/_y_
NUTRITIVE

VALUE

STUDIES

DAY 0

7FGP6f

W/FGP+METH
11/.,J
. i

21
I

t DAY PROTEIN
FREE

NITROGEN
BALANCE

PERIOD

BLOOD
SAMPLE

PERIOD
FIG I. Experimental
alternative
food (control
were not aware ofwhich
random.
Each nutritive
are designated.

TABLE
2
Representative
microfungal

amino
products

design of microfungal
tolerance
and nutritive
value studies.
Subjects
crossed over to the
or experimental)
after the initial 30 days ofthe tolerance
studies.
Physicians
and subjects
food was being taken during the 60-day periods,
and assignment
to treatment
groups was
value study began with a protein-free
day. Blood sampling
and nitrogen
balance
periods

acid composition
(gil 6 g N)

of the two

FGP(12)

Indispensable
Cystine
Isoleucine
Leucine
Lysine
Methionine
Phenylalanine
Threonine
Valine
Tyrosine
Tryptophan
Dispensable
Alanine
Arginine
Aspartic
Glutamic
Glycine
Histadine
Proline
Serine

acid
Acid

S Calculated
from a protein
43.7 g/lOO g dried product.

cium (Technicon
Method
no SE-COO3FJ4,
Technicon
Instrument
Corp, Tarrytown,
NY), phosphorus
(Technicon method
no SE4-0004FH4),
uric acid (16), creatmine (17), lactic acid dehydrogenase
(Worthington
LP
set SM520093,
Worthington
Biochemical
Corp, Freehold, NJ), alkaline
phosphatase
(Worthington
PNP set
SM5 10072),
amylase
(Harleco
amylase
reagent
set
64191, Harleco,
Division
of EM Industries
Inc, Gibbstown,
NJ)
(I 8), SOOT
(Worthington
GPT
set
SMS10031),
SGPT(Worthington
GPT set SM5 10046),
total protein
(Technicon
method
no SE4-0014FC4),
albumin
(Technicon
method
no SE4-0030FD4),
cholesterol (Enzymatic
serum cholesterol
determination
by
Boehringer-Mannheim
kit, 124087,
Boehringer-Mannheim
Biochemical
Co, Brookfield,
CT), triglyceride
(Worthington
triglyceride
reagent
set 27977).
In addition, a complete
blood count and urinalysis
were obtamed.
The urine collected
on each of the blood sampling days was tested for pH, glucose,
protein,
ketones,
and white and red blood cells.

0.73
4.31
6.96
7.24
1.92
3.73
5.24
5. 1 1
3.4 1
1.83
8.52
6.11
9.80
14.32
4.83
3.25
5.38
4.74
(N x 6.25)

PVP(I3)

0.77
4.12
6.49
6.33
1.52
5.78
4.14
4.93
3.49
1.88

Nutritive
5.78
6.10
8.09
10.82
4.37
1.89
3.81
3.94
content

of

value

studies

Subjects. Thirteen
healthy
male MIT students
were
recruited
for the FOP nutritive
value study and six for
the PVP study (Table 1). One subject (RS) participating
in the FGP nutritive
value study presented
aberrant
resultsand
therefore
was not included
in the calculation
of the mean values. Since variation
in his daily creatinine excretion
was excessive,
it is likely that he was not
adhering
strictly to the experimental
protocol.
Study design.
The 48-day
FGP study, was divided

Downloaded from www.ajcn.org by on November 11, 2010

1!

t#{188}6

MILK

288

UDALL

TABLE
3
Composition
ofthe
liquid formulas
used
and PVP nutritive
value studies (amounts
70-kg reference
man)

in the FGP
in g/day for

Diet
Ingredient
FOP

PVP
FOP
Dried skim milk
Polycosef
Dextri-Maltose*
CornOil
Avicel (Cellulose)lI
NaCl
Ca10 (OH)2 (PO41
K2HPO4
Water
Lemon juice
Pure vanilla
C To
enhance
mixing

Milk

PVP

46.4
45.7t
62.5
180.0
130.0
3.0
1.0
2.0
6.1
400.0

and

I 50.0
I 52.0
130.0
115.0
5.0
5.0
1.0
2.0
0. 1
2.0
3.5
6.1
500.0
450.0
5.0
10.0
palatability
properties,

PVP was combined

with 200 ml water boiled for 7 mm,
cooled, and water replaced
to 200 ml and blended
with
other ingredients.
FGP and skim milk powders
were
mixed directly with the other constituents.
t In the methionine
supplemented
FOP diet, L-methionine
was used at a level of 1.0 g/l#{174}g of FGP(lOO
g of protein).
t Ross Laboratories,
Columbus,
OH.
Mend Johnson
Laboratories,
Evansville,
IN.
H Donated
by FMC Corporation,
Food and Pharmaceutical
Products
Division,
Philadelphia,
PA.

AL

TABLE
4
Ingredients
of vitamin
and mineral
during the nutritive
value studies

supplements

given

One-a-Day
tablet (1 each day)
Vitamin
A 5000 IU
Vitamin
E 15 IU
Vitamin
C 60 mg
Folic acid 0.4 mg
Thiamin
1.5 mg
Riboflavin
1.7 mg
Niacin 20 mg
Vitamin
B 2 mg
Vitamin
B12 6 ig
Vitamin
D 400 IU
Pantothenic
acid 10 mg
Iron (elemental)
18 mg
Calcium
100 mg
Phosphorous
100 mg
Iodine
150 mEq
Magnesium
100 mg
Copper
2 mg
Zinc 15 mg
Trace mineral
capsule (1 each day)
MgO 398 mg Mg
CuCl2. 2 H2O 2 mg Cu
MnSO4.H2O
1.66 mg Mn
Na2 MoO4 .2H2O 0.083 mg Mo
Cr(SO4)3
15 H2O 0. 168 mg Cr
Na2SeO3 0.100 mg Se
AISO4.12H2O
1.61 mgAl
KI 0.29 ng I
Choline

bitartrate

(2 each

day)

250 mg choline/tablet

Energy need was determined

for each subject according to dietary history and an estimation
ofdaily activity.
This procedure
had been followed
previously,
usually
without
the need to adjust
energy
intake
during
the
conduct
of 60- to 80-day
studies.
Additional
energy
intake
was provided
when necessary
by protein-free
biscuits,
desserts,
and soft drinks.
Specimen
collection.
Four
venous
blood
samples
were taken during
the FGP study. The first two were
drawn just before and after the first 16-day period. The
other two were obtained
at the beginning
and end of
the third dietary
period (Figure
1). Two venous
blood
samples
were obtained
during the PVP balance
study,
at the beginning
and at the end of the 1 1-day period.
All samples
were drawn
between
8:00 and 9:00 AM,
after an overnight
fast. The same blood
tests were
performed
asdescribed
for the tolerance
studies. A urine
specimen
for routine
urinalysis
was also obtained
on
each ofthe
blood sampling
days.
A 24-h urine sample
was taken
1 wk before each
microfungal
study for determination
of urea nitrogen,
uric acid, and creatinine
(21). In addition,
complete
24h urine and fecal collections
were made throughout
the
study and tested for the same constituents
(21). Daring
the FGP study, fecal samples
were collected
and prepared as pooled samples
for the last 10 days ofeach
16day period
(Fig I). For the PVP study, fecal samples
were collected
for the last 7 days of the 1 1-day period.

Downloaded from www.ajcn.org by on November 11, 2010

into three
16-day dietary
periods:
FGP, cows milk
protein,
and FGP plus methionine.
Two dietary
sequences
were used: FGP-cows
milk-FGP
plus methionine or the reverse (Fig 1). Subjects
were enrolled
in
the study in two phases; four initially started the study,
followed
several
weeks
later by the remaining
nine
individuals.
The PVP study lasted 1 1 days; there was
no cows milk period (Fig 1). Each dietary
period was
begun after a 1-day protein-free
period
to accelerate
adaptation
to the dietary
period
(19). Subjects
were
weighed daily.
Diet. The sole source of dietary
protein
during
the
nutritive
value studies
was either
FGP, cows milk
protein,
or PVP. There were two experimental
periods in the FGP study in which participants
took either
FGP or FOP with 1% methionine
supplementation.
The ingredients
of the liquid formula
diet used in
these studies are shown in Table 3. The subjects
consumed their diets as four equal meals at 8 to 9:00 AM,
12 noon, 5:30 to 6:30, and 9 to 10:00 PM. A vitamin
tablet was given to each subject daily (One-A-Day
tablet, Miles Laboratory
Inc. Elkhart,
IN) and one mineral
tablet and two choline bitartrate
tablets (Plus Products,
Irvine, CA) were given to meet US National
Academy
of Sciences/National
Research
Council
recommendations (20) (Table 4).
In the FGP study, the total protein
intake during the
two experimental
phases (FGP and FOP plus methionine) and the control
protein
phase (cows milk) was
0.30 g protein/kg
body weight/day.
In the PVP study,
protein
intake was 0.35 g protein/kg
body weight/day.

ET

MICROFUNGAL
A fecal marker
(carmine
dye) was used to define the
collection
periods
(22). Aliquots
of the experimental
diets and of the pooled
fecal samples
were taken for
total nitrogen
analysis.
Assessment
of protein
quality
parameters
was made
according
to conventional
procedures
(23, 24). Thus,
total urinary
and fecal nitrogen
losses were corrected
for metabolic
urinary
and fecal nitrogen
and subtracted
from ingested
nitrogen.
Digestibility
was the percentage
of ingested
nitrogen
absorbed,
biological
value the percentage
of absorbed
nitrogen
retained,
and net protein
utilization
the percentage
of ingested
nitrogen
retained.

Results
Tolerance

siudies

buttocks
on the
1 5th day of what
later
proved
to be the experimental
period.
These
were described
by the examining
physician
as small,
nonpruritic,
evanescent
macules.
By the next morning
they had disappeared.
The rash reappeared
on the 20th day, and 3
days later she was told to discontinue
taking
the muffins.
The cutaneous
reaction
cleared
after 1 wk. She was started
on the muffins
again and completed
the 30-day
trial. On
the 30th day she developed
small,
pruritic,
maculopapular
lesions
on her left forearm,
right shoulder,
and right calf. At her request
she was dropped
from the study before
the
cross-over
to the control
muffin.
At the end of the PVP tolerance
trial, all
the diaries
were collected
and it was noted
that some individuals
had recorded
minor
symptoms
that they had not reported
to the
research
dietician
or physician.
Fifteen
of
the 50 subjects
mentioned
mild,
transient
gastrointestinal
symptoms
(nausea
and
bloating)
during
one of the two 30-day
penods.
Only six of these were receiving
the
experimental
muffins
at the time the cornplaint
was noted
in the diary.
Symptoms
were recorded
as early as the 1st day and as
late as the 30th day of the study periods.
Analysis
of blood and urine samples
obtamed
before and after the first feeding
period and at the end of the second
period
showed
that all values remained
within
normal limits for each of the 50 subjects.
Nutritive

value

studies

FGP.
The
weights
of the subjects
remained
relatively
constant
during the study.
The mean
weight
( SD) of subjects
on
the 1st day of the FGP, milk, and FOP plus
methionine
periods
were 69.9 8.7, 71.2
8.1, 70.8 7.4, respectively.
The
mean
weight
( SD) of subjects
on the last day
ofeach
study period was 70.5 8.7, 72.3
8.6, and 70.7 7.4, respectively.
During
the
first
dietary
period
two
subjects
were
dropped
from the study; one on the 5th day
and the other on the 1 lth day for reasons
not directly
related to ingestion
ofFGP.
One
of these individuals
had an acute infection;
the other subject
asked to discontinue
participation
because
of the demanding
regimen. The remaining
1 1 subjects
completed

Downloaded from www.ajcn.org by on November 11, 2010

One hundred
individuals
consumed
control
and
microfungal
protein-containing
cookies during the FOP study. None of these
individuals
had any gastrointestinal
reactions or skin rashes
that could
be ascribed
to ingesting
the experimental
material.
Minor complaints
recorded
in the diaries
of
several individuals
but not mentioned
to the
staff were judged
to be unrelated
to the
material
ingested.
With
one
exception,
no
significant
changes
were observed
in blood
tests performed
before and at the end of the control
and experimental
FOP feedings.
The exception was serum
cholesterol
that decreased
significantly
during
the 30-day
FOP period
from a prefeeding
mean value of 188 mg/dl
to a postfeeding
mean
value of I 75 mg/dl
(p < 0.001).
All but two of the 50 subjects
completed
the 67-day PVP study. One individual,
a 25yr-old woman,
complained
of severe headaches after 2 1 days of consuming
muffins.
These headaches
continued
for 4 days and
then subsided
spontaneously.
After the 7day muffin-free
interval
she was started
on
the other muffins.
This time she complained
ofabdominal
discomfort
and nausea
within
a day and her complaints
continued
for 2
wk. At this time she was dropped
from the
study. The relationship
of her symptoms
to
the two dietary
phases was known
only after
she stopped
eating the muffins
and the study
code was broken.
It turned
out that the first
phase was the control
and the second
the
experimental
period.
Another
female
subject
noted
minor
rashes
on her right elbow,
lower leg, and

289

FOOD

290

UDALL

AL

TABLE
6
BUN (mg N/100
ml) ofyoung
nutritive
value study

5
of the nutritive

value

of FGP

in young

adult

Protein
.

VD
EA
RC
KD
MC
RM
DR
WR
Wi
DD

Milk

BVt

80
79
57
78
78
75
91
73
95
69

83
107
74
75
99
80
90
87
52
89

66
85
42
59
77
60
82
64
49
61

84
15

65
14

Mean
78
SD
11
a Digestibility.

t Biological

FGP+
,,,thioni

FOP

Subject

NPUt

B/

NPU

91
98
87
94
92
88
101
97
100
99

93
98
93
74
99
81
81
93
62
77

85
96
81
70
91
71
82
90
62
76

79 107 85
80 113 90
63
9057
78
9473
81
99 80
81
9275
788264
82
90 74
91
5550
81
96 78

95 85
5 12

80
11

79
7

BV

#{149}

NPU

9273
16 12

value.
t Net protein
utilization.
For calculation
of BV and NPU,
obligatory
urine
and fecal N losses were taken to be 37.2 and 8.8 mg N/
kg body wt/day
(35).

in the FGP
Final

Initial
value

Subject

FOP

MIkI

value
FGP+
methionine

VD
EA
RC
KD
MC
RM
DR
WR
Wi
DD

1 5.0
16.0
17.0
14.0
16.0
16.0
11.0
19.0
13.0
14.0

6.0
4.0
5.0
6.0
7.0
4.0
5.0
5.0
6.0
4.0

4.0
8.0
4.0
3.0
8.0
6.0
7.0
4.0
6.0
6.0

4.0
2.0
5.0
7.0
4.0
4.0
4.0
3.0
6.0
6.0

Mean
SD

15.1
2.2

5.2
1.0

5.6
1.8

4.5
1.5

nutritive

value

TABLE
Estimates
men

TABLE
Estimates
men

men

7
ofthe

of PVP

in young

Subject

BVt

NPU*

SL
DB
EH
AN
JK
CC

87
80
86
77
78
76

55
70
79
37
93
69

48
56
68
29
73
52

81
5

67
19

54
16

Mean
SD
Digestibility.

adult

t Biological

value.
:t Net protein
utilization.
Values calculated
on 3-day sample.
For calculation
of BV and NPU, obligatory
urine
and fecal N losses were taken to be 37.2 and 8.8 mg N/
kg body wt/day
(35).

were no significant
changes
in body weights
of these subjects
during
the study interval.
The weights
( SD) on the 1st and last day
ofthe
study were 66.4 5.7 and 65.8 5.9
kg, respectively.
One subject
(CC), because
of nausea,
asked to terminate
the study on
the 9th day. The other
five subjects
cornpleted
the entire
study,
and none
of the
remaining
individuals
complained
of any
symptoms
to the supervising
physician
or
dietician.
Two subjects,
however,
recorded
in their diaries that they had occasional
mild
nausea.
As can be noted
in Table
7, the mean
digestibility,
biological
value,
and net protein utilization
were 8 1, 67, and 54, respectively; values not significantly
different
from

Downloaded from www.ajcn.org by on November 11, 2010

the first, second,

and third
study
periods
with no significant
complaints.
The results
of the calculations
of digestibility, biological
value, and net protein
utilization
are summarized
in Table
5. The
mean digestibility,
biological
value, and net
protein
utilization
for the group
of 10 mdividuals
taking
0.30 g of FOP per kg body
weight/day
were 78, 84, and 65, respectively.
The mean digestibility,
biological
value, and
net protein
utilization
for the group
of 10
individuals
on 0.30 g/kg/day
of the skim
milk formula
were 95, 85, and 80, respectively.
The differences
in digestibility
and
net protein
utilization
were statistically
significant
by paired t test analysis
(p < 0.001),
but the biological
values
were similar.
As
also shown
in Table
5, methionine
supplementation
did not affect
digestibility,
but
resulted
in significantly
improved
biological
value and net protein
utilization
(p < 0.05).
With the exception
of the BUN, all blood
and urinalysis
tests remained
normal
during
the three
phases
of study.
The BUN
decreased
significantly
(Table
6) in all three
dietary
periods
(p < 0.00 1) compared
with
initial values.
PVP. Five of the six subjects
completed
the 1 1-day nitrogen
balance
study and there

ET

MICROFUNGAL

those
of FOP.
However,
both
biological
value
and net protein
utilization
of PVP
were significantly
lower (p < 0.02 and <
0.05, respectively)
by a two sample
I test
when compared
to FOP supplemented
with
methionine.
Mean serum
BUN in blood obtained
before the start of the PVP nutritive
value
study was 1 2.8 mg N/tOO
ml and the final
BUN was 6.2 mg N/ 100 ml (p < 0.0 1 ). Mean
serum uric acid of subjects
before the study
was5.5
l.2mg/l00ml(SD)and9.2
4 mg/ml
at completion
of the study (p <
0.001).
The other
initial
blood
and urine
tests were not significantly
different
from
final values.

The two microfungal

foods studied
were
well tolerated.
None of the 1 13 individuals
who consumed
the FOP product
had even a
suggestion
ofan adverse
reaction.
Two of 56
subjects
taking
PVP had mild
symptoms
that may have been due to the supplement.
Adverse
reactions
to accepted
foods
have
been estimated
to occur
in 1 to 4% of the
population
(25). Therefore,
we conclude
that
the two microfungal
products
we studied
are
well tolerated
and that the likelihood
of adverse reactions
to these products
is no greater
than with many common
foods.
There were changes
in several serum components
noted during
the tolerance
and nutritive
value studies.
Cholesterol
decreased
slightly but significantly
during
the FOP tolerance
study although
not during
the PVP
tolerance
study.
Animals
fed FOP have a
decrease
in serum
cholesterol
(26-29).
This
effect may result when cholesterol
excreted
as bile acids into the small intestine
is adsorbed to the FOP, blocking
reabsorption
of
the bile acid and decreasing
the cholesterol
pool (26).
Nineteen
subjects
were monitored
more
closely but for shorter
periods
oftime
during
the nutritive
value studies.
BUN decreased
in the FOP and PVP nutritive
value studies
as expected
because,
by design,
the individuals participating
in these studies
were all in
negative
nitrogen
balance.
Serum
uric acid increased
significantly
with the intake ofthe
PVP during
the nutri-

291

tive value study from an initial value of 5.5

mg/dl to a final value of9.2 mg/dl.
This just
exceeds
the upper limit for the normal
adult
male (3.9 to 9.0 mg/dl).
PVP contains
8%
nucleic acid. Unlike
the FOP it is not nucleic
acid reduced.
Therefore,
a 70-g reference
subject
consuming
0.35 g of PVP
body
weight/day
would
ingest 24.5 g of PVP/day
which is 8% nucleic
acid. That individuals
daily intake
of nucleic
acid would
be 2.0 g/
day, the maximum
recommended
daily intake of nucleic
acid (30).
The daily consumption
of additional
nucleic acid by subjects
during
the FOP nutritive value study was much less, and as anticipated,
did not significantly
increase
serum
uric acid. These results confirm
the need to
reduce the nucleic
acid content
of microfungal proteins,
ifthey
are to be used as a major
protein
source
in human
diets.
It is not
necessary,
however,
if they are to be used as
a food additive,
as long as the additional
daily nucleic
acid from this source
does not
exceed
2.0 g (30). This limit will maintain
serum
levels of uric acid within
acceptable
limits for normal
subjects
and minimize
the
risk of deposition
of urate crystals
in kidneys, joints,
and other tissues.
The
digestibility
of both
microfungal
products
was approximately
83% that of
casein.
This lower value may be due to the
poorly digestible
cell walls (3 1 ). The digestibility values are similar
to those obtained
by
others for yeast, bacterial,
and algal proteins
(2, 3, 32, 33). The biological
value and net
protein
utilization
data for both microfungal
foods are also similar
to those obtained
by
others
in human
balance
studies
of microbial proteins
(2, 3, 32-34).
The results
of these studies
indicate
that
both microfungal
foods are acceptable
and
nutritious
for human
diets. FOP can be used
as a major protein
source in the form tested.
PvP is suitable
as a food additive,
but as a
major
protein
source
it would
need to be
processed
to reduce the nucleic acid content.

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ET