INTRODUCTION

• Document Type: CREST
• Collection: General CIA Records
• Document Number (FOIA) /ESDN (CREST): CIA-RDP79-00798A000400090014-0
• Release Decision: RIFPUB
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• Document Creation Date: December 27, 2016
• Document Release Date: January 14, 2013
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mow' Nue
INTRODUCTION
[Introduction written by G. K. Skryabin, Corresponding Member
of the USSR Academy of Sciences, and A. A. Pokrovskiy, Academi-
cian of the USSR Academy of Medical Sciences; Moscow, Mediko-
Biologicheskiye Issledovaniya Uglevodorodnykh Drozhzhey, Russian,
1972, pp 5-8_
The Directives of the 24th CPSU Congress provided for
scientifically sound prospects for further improvements in the
welfare and nutrition structure of the Soviet people. Proposals
in this area are directed toward further improvement in the health
conditions of the population, increase in the efficiency and
lengthening of the period of active life of human beings.
In solving the food problem, the socialist society bases
itself on a progressive and vital concept ensuring a planned pro-
duction of foodstuffs in the proportions, amounts, and conditions
needed by the society. This concept is based on a single plan
of the development of socialist economy whose main goals are to
ensure the health and happiness of mankind.
In predicting the routes for the solution of the food pro-
blem for the following decades, it should be foreseen that the
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Noe
natural productivity of our biosphere based on the accumulation
of solar energy, rationally adjusted by the methods of agricul-
tural production, and strengthened by achievements of the scien-
tific and technological progress, first of all, by chemistry and
microbiology, will continue to be the main source of nutrition
for the population of our planet. This means that the main forces
of science must be concentrated on how to make the agricultural
production more profitable and productive, on how to increase the
nutritional utilization of the overall productivity of the bio-
sphere, and on how to strengthen more reliably the feed resources
of animal husbandry.
At the same time, if we consider the food problem on a
global scale, it becomes evident that the use of the traditional
methods of food production will not sufficiently solve the food
problem of the world in the near future. Unfortunately, the con-
siderable progress made in the agricultural production in the
last decades does not make it possible to solve the problem of the
protein crisis. It is also obvious that a considerable shortage
of proteins in the diets of human beings which was termed as the
protein crisis in numerous UN documents is a serious detriment
to the health of hundreds of millions of people. Children of
early ages are the most sensitive to the shortage of proteins.
It is generally known that a chronic protein deficiency during
the first years of human life results in a lagging not only in
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the physical, but also in the intellectual development, and leaves
its aftereffects for the rest of the person's life.
In solving the strategic problems of the food policy of
the future, the attention of the scientists of technologically
developed countries has been increasingly concentrated on the
possibility of using proteins of unicellular organisms which
are natural concentrates of proteins.
The considerable materials accumulated by the Soviet and
foreign researchers leave no doubt that the methods of micro-
biological synthesis are exceptionally promising in -Ole solving
of problems of increasing the overall protein resources of our
planet. The possibility of using many different organic com-
pounds for growing microorganisms, including petroleum hydrocar-
bons, natural gas, wastes of various chemical industries, as
well as an intensive development of methods for producing bio-
masses pf these microorganisms, poses the problem of their wide
utilization in solving the problem of the increasing protein
shortage in the world.
Here, we must stress the rapid pace and considerable pro-
gress in the development of the studies on the problem of pro-
teins of unicellular organisms in the USSR. In the Soviet Union,
large-scale investigations are carried out on the above-mentioned
potential sources of microbiological proteins, including yeasts,
bacteria, and algae. The progress of Soviet specialists was
particularly significant in the area of studies on yeasts devel-
ing on hydrocarbons.
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'i4110e
The Soviet Union is the first country to organize a large-
scale production of nutrient yeasts on the basis of petroleum
hydrocarbons. At the present time, the Soviet Union is build-
ing large plants for the production of protein concentrates of
microbic origin.
The significance of these new means of producing proteins
cannot be overemphasized. Calculations show that only 2 percent
of the presently available petroleum is sufficient for the pro-
duction of 25-30 million tons of yeast proteins capable of feed-
ing 2 billion people. It is natural that the introduction of
the fundamentally new method of obtaining protein substances and
the discussion of the possibility of using the proteins of uni-
cellular organisms as nutrient products for people and animals
will require thorough medico-biological studies.
This book summarizes the results of the first stage of
medico-biological studies on hydrocarbon yeasts which were car-
ried out, chiefly, at the Institute of Nutrition of the USSR
Academy of Medical Sciences.
The investigations reported in this book cover a rela-
tively long period of time: from 1964 to 1970. Naturally, in
the course of these studies both the general ideas of the re-
searchers of BVK (protein-vitamin concentrates) and the research
methods were improved gradually. Thus, the papers published
here reflect, to a certain degree, the evolution of the in-
vestigation process.
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INV N409
The introductory part of the book treats the substantia-
tion of the statement of the problem and the selection of the
study methods.
The second section presents a series of experimental
studies on the biological value and harmlessness of natural (BVK)
and extracted (biological oil-seed meal) yeasts grown on media
containing petroleum n-alkanes as a source of energy.
The third section treats the biological value and harm-
lessness of animal husbandry products obtained as a result of
using BVK in feeding agricultural animals.
The fourth section describes the results of control ob-
servations on groups of volunteers from the members of the In-
stitute of Nutrition of the USSR Academy of Medical Sciences and
the All-Union Scientific Research Institute of Protein Synthesis
of the Glavmikrobioprom [expansion unknown] who were receiving
meat products in their rations from animals fed with the addi-
tion of BVK regularly for many months.
The last part of the book gives a brief analysis of the
obtained results and an attempt at generalizations and conclu-
sions regarding the possibility of practical utilization of BVK
in animal farming. On the whole, the results of the medico-
,biological studies on the problem of BVK have served as an ade-
quate basis for important decisions on the use of this new and
very promising source of nutrient yeast for animal husbandry.
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The accumulated accumulated data will, undoubtedly, serve the purpose
of further development of investigations on the problem of ra-
tional utilization of proteins of various species of unicellular
organisms directly in human diets.
In conclusion, we would like to mention that, in spite
of a large number of authors, the book is of a monographic nature
because of its single plan of methodological approaches and the
consistency of presentation. In essence, the materials presented
in this book are the first broad generalization of the results
of medico-biological studies on this problem, and they can serve
as an important guide for a basic evaluation of the prospects
for the development of a new branch of technological microbiology
Or????=11
the production of protein products on the basis of micro-
biological synthesis.
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Nip" Now
RESULTS OF MEDICO-BIOLOGICAL TESTS ON HYDROCARBON YEASTS
[Article by A. A. Pokrovskiy, Institute of Nutrition, USSR Acad-
emy of Medical Sciences, Moscow; Moscow, M2dik2=1.31212qj.cheskive
Russian 1972 pp 449-463]
This book summarizes the results of six years of medico-
biological studies which were conducted by the Institute of Nutri-
tion, USSR Academy of Medical Sciences, (from 1964 to 1970) for
evaluating the quality of yeasts grown on a base of the paraffin
fraction of petroleum whose dry biomass was called BVK [protein
-
vitamin concentrate]. This book is somewhat fragmentary in nature
And has been written by a large group of authors. Nevertheless,
it is based on one general plan and has a single purpose. The
book treats two most important problems which substantially de-
termine the possibility of the development of a new branch of
microbiological industry: firstly, the establishment of the bio-
logical value and harmlessness of hydrocarbon yeasts and, second-
lyithe determination of their complete harmlessness when used in
animal husbandry.
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411fte 'Nrrof
The great theoretical and practical significance of this
new source of protein stems, first of all, from the use of a new
energy source, petroleum hydrocarbons, for the biosynthesis of
protein and other nutritional substances. Petroleum hydrocarbons
have never been used before for this purpose in the process of
indu-trial biosynthesis. In connection with the huge reserves
of Petroleum that have already been discovered on the earth, the
Poueibilities of its utilization for increasing the protein re-
soutces of our country have, naturally, attracted the attention
not only of the representatives of the oil industry, but also of
many scientific research laboratories in the world.
A very important step was the interpretation of the bio-
.
chokical mechanisms of the first stages of the utilization of the
enf,gy of paraffins by yeasts and the discovery of alkane dehy-
dro7 enase, a key enzyme which makes it possible for microorgan-
leak to use the energy of hydrocarbons of normal series.
Considerable progress in the development of the production
te0mology of new kinds of yeast in our country has created real
Pn?equisites for organizing their large-scale production. Scien-
tirsi,s working in the field of animal husbandry, as a result of
thw:x long-range investigations conducted simultaneously with
mod:co-biological studies, gave identical answers regarding the
Poibility of using these yeasts as sources of additional pro-
toiks in the nutrition of some species of agricultural animals.
At The same time, the problem of the possibility of a wide use
?f 'i.ydrocarbon yeasts in animal husbandry has been left open.
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law NINO'
The fundamentally new product, its use in industry as
the main energy source of the paraffin fraction of petroleum, and
the presence of residual hydrocarbons in it, as well as the pos-
sibility of accumulation of unusual products of their transforma-
tion, made it necessary to subject BVK to detailed medical studies
directed, first of all toward revealing the possibility of negative
biological effects. For this, not only the usual methods were
used for revealing an acute chronic toxicity and cancerogenic,
mutagenic, and teratogenic effects, but also scrupulous studies
were made on the effect of BVK on the state of the basic physio-
logical systems of the organism, the progress of metabolic process,
and the state of enzymic constellation, as well as on the routes
of transformation of residual hydrocarbon.
The medico-biological studies were done on the basis of a
single complex plan and unified schemes and methods of study ap-
proved by the Ministry of Health.
Studies of BVK were conducted on tens of thousands of ex-
perimental animals, including several populations of white rats,
dogs, rabbits, guinea pigs, and monkeys.
Various specimens of hydrocarbon yeasts were studied. The
tests revealed a clear tendency toward improvement in their bio-
logical qualities as the technological process was improved. For
example, during the first stages of work, yeast grown on petro-
leum paraffins with a high content (up to 2 percent) of residual
hydrocarbons was used. The experimental rats which received such
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Yeast had sufficiently marked pathological disturbances, partie-
nlarly in the liver and the kidneys, which were beyond the limits
qf changes resulting from deficiencies of individual irreplace-
'able amino acids.
Subsequent studies were carried out with improved speci-
likens of yeasts whose content of residual hydrocarbons did not
6xceed 0.6 percent, then 0.5 percent, and finally, 0.1 percent.
1? was established that the proteins of hydrocarbon yeasts were
assimilated by animals quite satisfactorily at a definite quota
tncluded in their rations and ensured normal growth and develop-
kent both in experimental and agricultural animals.
In metabolic tests, the processes of assimilating protein
qf hydrocarbon yeast were studied in detail, particularly the in-
flexes of nitrogen balance, digestibility, assimilability of pro-
and others.
In numerous series with replacement of various amounts of
brotein in food rations with yeast protein, the biological pro-
beities of BVK proteins were compared with known rich proteins
(casein a protein complex of rich vivarium rations), as well
s proteins of other kinds of nutrient yeasts. As a result of
this, a fundamentally important conclusion was made that the best
kpecimens of hydrocarbon yeasts were not any inferior to other
kinds of nutrient yeasts, such as sulfite and hydrolytic yeasts,
in the indexes of their effect on the development of growing
gnimals.
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.mivr 'gar
At the same time, it was shown that in the cases when
yeasts were the only source of protein, regardless of the content
of residual hydrocarbons in them, the experimental animals devel-
oped considerable disturbances in their organs which were charac-
teristic chiefly of a deficiency in sulfur-containing amino acids.
The degree of these disturbances decreased when the percentage
of the yeast in the rations was reduced or when calculated amounts
of amino acid (methionine) limiting the biological values of the
yeast were added. Then, the degree of balance of the amino
acid composition of the ration as a whole improved suipstantially.
When hydrocarbon yeasts were used in amounts not exceed-
ing 25 percent (on conversion to protein content) these changes
were minimal, and when 10-15 percent of yeast was added, no changes
were apparent in any of the cases.
Thus, hydrocarbon yeasts corresponding to definite condi-
tions can be evaluated as a quite satisfactory source of addition-
al nutrient protein.
Special experiments were conducted in order to studylin
detail, the fate of the components of the lipid fraction of hydro-
carbon yeasts. Its fatty acid composition, as well as the phos-
pholipid and sterid components, were studied. It was established
that their lipid fraction contained a relatively large amount of
fatty acids with odd numbers of carbon atoms (ZhKN) unusual for
natural foodstuffs. It was also established that these acids are
also capable of being accumulated by the tissues of experimental
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vir
and agricultural animals; the degree of ZhKN accumulation was
higher when the yeast quota in the food rations and the lipid
content in them were greater.
Due to this, the fatty acid composition of the triglycer-
ide fraction could change substantially in experimental animals,
and a considerable percentage of ZhKN accumulated in the fat,
particularly in the internal fat.
Taking into consideration the fundamental importance of
determining the possibility of transition of hydrocarbon into
animal tissues and the fact that this problem had not been studied
to any extent, special attention was given to developing methods
for the determination of residual hydrocarbons in foodstuffs.
It was shown that the methods of extracting hydrocarbons with
hexane which had been accepted as standard for analyzing hydro-
carbon yeasts were completely unsuitable for analyzing foodstuffs.
It was also found that the method of distilling hydrocarbons with
phenol developed by a group of researchers in the Institute of
Nutrition of the USSR Academy of Medical Sciences was practically
unsuitable for this purpose because of its low sensitivity. Sat-
isfactory results were obtained on the basis of extracting food-
stuffs with a mixture of 'chloroform .with methanol (2:1), two-stage
extraction with hexane, subsequent separation of the extract by
means of chromatography in a thin layer of silica gel, and, fur-
ther, by means of a gasochromatographic analysis of hydrocarbon
components.
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Nir 4.0
These methods provided a basis for revealing the fact of
the transition of residual hydrocarbon yeasts into the tissues
of organisms of experimental and agricultural animals and for the
study of the dynamics of their elimination from the organisms.
It was established that the maximum accumulation and the longest
retention of hydrocarbons in the organisms of animals are observed
in the fat tissues and, particularly, in the so-called internal
fat.
A special series of experiments was inteftded for studying
hydrocarbon metabolism by using carbon-labeled octadeCane which
constitutes about 15 percent of all paraffins contained in yeasts.
It was shown that this hydrocarbon was rapidly absorbed and dis-
tributed in the tissues of experimental animals when it was intro-
duced into their gastrointestinal tracts. When octadecane was
introduced once in a tracing dose, considerable quantities of it
were detected in the tissues during the first 24 hours. By the
15th day, its concentration dropped sharply, and after two months
the radioactive labeling was found only as traces, chiefly, in
the fat tissues. At the same time, it was established that octa-
decane was gradually metabolizing in the organism and the radio-
active labels moved to the fraction of fatty acids. Thus, the
concept of the extreme stability of hydrocarbons in the tissues
of the organism must be supplemented with the idea of their pos-
sible oxidation and transformation into corresponding fatty acids,
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'4wler
The above-mentioned data on the accumulation of lipids and
hydrocarbons in animal tissues became a basis for the decision
of the Board of the USSR Ministry of Health to allow the use of
BVK in meat husbandry (pigs, horned cattle, broilers) only under
the condition of the maximum permissible content of residual hy-
drocarbons in the yeast (0.1 percent) and the necessity of exclud-
ing BVK from the rations of agricultural animals and fowl at a
specified time before they are slaughtered. Let us note that this
conclusion is, to a considerable degree, in agreement with later
publications issued by the research laboratories of the "British
Petroleum Company," where agricultural animals were fed with
yeast in which the residual amount of hydrocarbons was also less
than 0.1 percent (Lovellin, Shekledi).
Special studies dealt with the development of methods and
attempts to detect therepresentatives of the aflatoxin family
in the composition of hydrocarbon yeasts. As is known, they are
quite widespread metabolites of a number of microscopic fungi
possessing a high degree of hepatotropic toxicity and cancerogen-
icity. By applying the extracting of yeasts and animal tissues
with Folch's mixture with subsequent thin-layered chromatography
and a study of the chromatograms by the methods of luminescence
and spectroscopic analyses made it possible to make a reliable
conclusion that there were no aflatoxins in the submitted yeast
specimens. At the same time, when some specimens of yeast were
studied by these methods, the chromatograms revealed the
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bresence of some substances with a characteristic yellow-green
fluorescence which must be studied further.
In the medico-biological studies, attention was concen-
Erated, chiefly, on studying foodstuffs obtained from agricultur-
al animals which received BVK in their food. A wide spectrum of
foodstuffs were studied: flesh of pigs, subcutaneous and internal
fat, various kinds of subproducts, flesh of broilers, milk, butter,
cottage cheese, chicken eggs, and others.
Detailed studies of the hygienic, physicochemical, and
brganoleptic properties did not reveal any positive changes in
these products. Evaluating them by a number of tasters did not
reveal any differences from the corresponding control products.
Por example, the meat of the experimental animal was sufficiently
'bander, aromatic, and tasty. Neither the meat itself nor the
boullions obtained on its basis had any unusual odors or tastes.
Chemical studies on the composition of meat products obtained
from agricultural animals fed according to the recommendations
,t)f the Board of Ministry of Health did not reveal any positive
changes in their chemical composition.
Long-range experiments on the effects of these meat pro-
ducts (including subproducts) on the development of several gen-
..rations of rats did not reveal any deviations in their behavior
Ind in the dynamics of their growth and development, as well as
in the state of their protein, lipid, and carbon matabolism, and
.he activity of the studied enzymic systems in the condition of
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a number of hematological, morphological, and histochemical in-
dexes. The definitely positive results obtained in the detailed
experimental studies of meat products made it possible to conduct
large-scale control observations of the effects of regular con-
sumption of experimental meat products on large groups of volun-
teers from among the members of the Institute of Nutrition of the
USSR Academy of Medical Sciences. The observations were carried
out twice in the course of six months on groups of over 100 per-
sons. These studies confirmed that these products were suffi-
ciently tasty and highly nutritious, and that there were no un-
favorable effects resulting from their consumption. These con-
clusions are based on varied and systematic clinical, hematologi-
cal and biochemical studies on the above groups of volunteers.
Particularly thorough studies were carried out on assess-
ing the biological value and harmlessness of milk, dairy products
and eggs from cows and chickens fed with the addition of BVK.
The point is that the interruption in feeding with BVK in order
to free the organisms of the animals from unusual substances
(paraffins, ZhKN) does not serve its goal in the case of lactation
of the cows or the production of eggs by laying hens.
Long-range studies of these products yielded very favor-
able results with respect to the growth and development of the
experimental animals. No differences were detected between the
experimental and control groups of rats, as well as in the mor-
phological and hematological indexes, enzymic activity in the
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tissues of liver and blood, basic indexes of protein and hydro-
carbon metabolism, and so on. At the same time, tendencies toward
an increase of fli-lipiproteid fraction, cholesterin and general
lipids of the blood serum were revealed during later observation
periods (6-7 months) in animals receiving experimental products.
The observed changes in some indexes of lipid metabolism result-
ing from the consumption of dairy products and eggs, since these
types of products are used daily and widely by all strata of the
population in their nutrition, particularly by children and elder-
ly people, make it necessary to have a more careful approach to
the problem of whether or not it is expedient to use BVK as an
addition to the food of lactating cows and laying hens. This con-
clusion is substantiated even more by the fact that there are
widespread disturbances in lipid metabolism among middle-aged and
elderly persons.
Summing up the results of medico-biological studies, it
should be mentioned that hydrocarbon yeasts may be considered as
a very promising source of additional proteins. Many problems
which existed at first in this connection have now been eliminated,
and representatives of various branches of biology have arrived
at the same conclusions.
The most important results of medico-biological studies
on the problem of BVK may be presented briefly in the form of
answers to the questions which were posed to the researchers of
the USSR Ministry of Health.
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Questi
a: Are cancerogenic
position of r
qrocarbon yeasts?
Answt.,
ticularly it ,
the paraffin
leukosogenic
cocanceroge
Quesil
Answt.
substances present
in the corn-
If the raw materials are carefully selected, par-
7drocarbons of the cyclic series are eliminated from
raction, there are positively no cancerogenic or
Uects from hydrocarbon yeasts. Also, no marked
effect of these yeasts has been detected.
,n3 Do hydrocarbon yeasts have a teratogenic effect?
on experime No teratogenic effects were ever found in studies
/,)
A
Quesi animals or in large-scale agricultural experiments.
Answc,
Quest
yeasts and w4,
these yeast4
Answc,
pertmental
possible to
these yeastt-,
of nitrogen 4
the 2roteim
tent: of me.t1
ical value
introduced
amoumt of mt.
Do hydrocarbon yeasts have a mutagenic effect?
No mutagenic effects were revealed in using BVK.
,n: What is the biological value of hydrocarbon
t are the indexes of nitrogen retention in using
feeding various species of animals?
The results of experiments on many species of ex-
agricultural animals, including monkeys, make it
ye an identical answer: the protein components of
re assimilated quite satisfactorily. The retention
in agreement with the amino acid composition of
E a given kind of yeasts. A comparatively low con-
-nine limits,
is the primary
to a certain degree, their biolog-
reason to limit the amount of yeasts
o the rations or to enrich them with a calculated
itonine.
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--wpw Now
Question: Have definite differences been revealed be-
tween the bi:-,logical value of BVK and other kinds of nutrient
yeasts?
Answe--$ When experimental animals are fed with the addi-
tion of hydr;-) carbon yeasts according to the medical requirements
stated in thoa resolutions of the Board of the USSR Ministry of
Health of 198, their effect on the development of all species
of the studi animals does not differ in any respect.. from other
types of nut.:.-ient yeasts.
Quest ions Are there any changes in the quality of the
products obt?Iiined from cattle fed with the addition of hydrocar-
bon yeasts?
Answer: No definite changes have been detected in the
quality of rife-at products when studies were carried out on a wide
spectrum of their hygienic, physiocochemical and organoleptic
properties.
Quest 'ion: Have the limitations in the use of hydrocarbon
yeasts for c*ttle feeding been substantiated?
Answer's Yes, they have been substantiated. In calculat-
ing rational doses of BVK (with respect to the protein
in the ratio;10), it is not expedient to increase its amount above
10-15 percerVt. When the yeast dose is increased above 25-30 per-
cent, there *re definite deviations from the normal development
of experimental animals (some lag in gaining weight, sometimes
fat infiltr
tion of liver, and so on).
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.fift"
Nu"
'411eitions What are .t.e reasons for these limitations which
workers?
('111,7erg Apart from t-r.oa above-mentioned nonoptimal amino
ale! '"P? ition, the followl.-Ag facts make it necessary to exer-
-'4.tqess:
Lhe presence of a cert,ain amount of residual petroleum
jbatis in the compositi_n of hydrocarbon yeasts; it should
'c'1"lidered that resith1 petroleum hydrocarbons are iden-
vis
part;
st31
-drocarbons of some plant products which are mostly of
ec4lar weight and are, chiefly, contained in vegetable
-.26 C34);
he presence of rather uncommon lipids in paraffin yeasts,
? driy, a relatively h.-2?h content of fatty acids with odd
?!arbon atoms, compr--1.-Ation of sterols unusual for animal
alid so on;
linusually high content of purine and pyrimidine bases
/eallts, which, however, is characteristic of the absolute
(31- microorganisms and which is of utmost importance in
-ttl/Ls of using them in diets of human beings.
''.1"tions Ca..1 normal r)araffinsbe absorbed in the gastro-
tract?
(151'er: Yes, they can. Studies with direct determination
tilis in yeasts and Wsues of experimental animals by the
11 gas chromatography,
as well as experiments with paraf-
d for carbon, made it possible to reveal their ability
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Aloe *NW'
to penetrate rapidly through the intestinal mucosa and be retained
in the tissues of the organism.
Question: In what tissues does maximum accumulation of
residual hydrocarbons take place and where are they retained for
long periods of time?
Answer: These tissues are the subcutaneous and, particu-
larly, internal fat. It is here that hydrocarbons accumulate the
most and are ratained for comparatively long periods of time.
After these facts had been established, it became possible to sug-
gest methods for effective control over the quality of meat pro-
ducts obtained from animals fed with the addition of hydrocarbon
yeasts.
Question: Can fatty acids with odd numbers of carbon atoms
accumulate in the organism tissues?
Answer: Yes, they can. It has been proven that some changes
in the fatty acid composition of the organism tissues are observed
when considerable amounts of BVK are used in the diets of labora-
tory and agricultural animals.
Questions In what tissues the accumulation of these acids
is particularly expressed?
Answer: The largest amounts of fatty acids with odd num-
bers of carbon atoms were found in the internal fat, and short-
term observations also revealed them in the liver.
Question: Can residual hydrocarbons and fatty acids with
odd numbers of carbon atoms be metabolized in the organism of
animals?
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Answer-; Yes, they can. According to the available data,
fatty acids TAT:_th odd numbers of carbon atoms are subjected to suf-
ficiently int4.,:!nsive processes of biological oxidation. However,
some fermenta
ion stages of this process differ from the trans-
formation proesses of fatty acids with even numbers of carbon
?
atoms which a
e usual for the organism. As for n-alkanes, our
experiments w.:-th labeled octadecane yielded convincing proofs of
the possibiliy of a gradual transformation of alkanes into the
corresponding fatty acids.
Questif,in: Does BVK contain substances of the furocumarine
series of the type of aflatoxins?
Answer$ The BVK specimens submitted for study were com-
pletely free from such substances,
Such TAmre the results of the medico-biological studies
which, undoub4-edly, characterize the new type of yeasts as an in-
teresting and promising source of additional proteins and, to a
considerable degree, map out the routes for their effective and
safe applicatlon in agricultural production.
The results of these investigations, together with the pub-
lished data, can make it possible to give an unambiguous answer
to the alternative question: which course of study should be pre-
ferred in the immediate future for the investigation of single-
cell organisms -- a direct one or through an animal organism.
In comparing the peculiarities of the production of hydro-
carbon yeasts, it is necessary to consider the fundamental differences
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'411W
both in the source ed for the biosynthesis of energy, and in
the food chains con:-Jacting them with man, on the one hand, and
proteins in the cordriary agricultural production, on the other
(diagram).
Key: 1) Man 5)
2) products of animal 6)
origin(milk,meat,eggs) 7)
3) Agricultural animals 8)
4) Solar energy 9)
Plant biomass
Biomass of yeast cells
CO2 of air
Soil substances, fertilizers
Nutrient medium (hydrocarbons+
t NH4 t mineral salts)
In one instance, the role of an intermediate link in the
chain of the assimilation of food substances from the environ-
ment is played by multicellular plant organisms equipped with a
complex mechanism of selective sorption of only those substances
which are necessary for the organism.and having a considerably
more expressed, genetics-dependent stability of their chemical
composition. In another instance, the intermediate link in the
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"gel"
food chain is represented by rapidly developing cultures of uni-
cellular organisms which are much more capable of absorbing for-
eign substances and admixtures from the nutrient medium. In this
case, the introduction of petroleum hydrocarbons into nutrient
media and the possibility of their accumulation in the cellular
biomass caused particular concern. However, apart from hydrocar-
bons, in evaluating the potential harmfullness of hydrocarbon
yeasts, it is also necessary to consider the possibility of the
presence of other substances unusual for natural foodstuffs in
them, for example, unusual components of the lipid fraction (ste-
rols, ZhKN, and the like), unusual amino acids (for example, di-
aminopimelic acid, D-optical isomers of amino acids, and so on),
amino sugars and peptides, particularly in the composition of the
cellular membrane, an extremely high content of nucleic acids,
biologically active compounds of the type of mycotoxins and anti-
biotics, and others so far unknown toxic components. All these
facts, as well as a considerable changeability of the properties
of microorganisms and the insufficient understanding of the laws
of this changeability in the process of the mass production of
the biomass, do not leave any doubt about prematureness and a con-
siderable risk of a wide introduction of proteins of microbiologi-
cal origin directly into the diet of human beings at the present
time.
In making prognoses for the means of solving the food pro-
blem for the next decades, it should be expected that natural
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productivity of of the biosphere based on the accumulation of solar
energy, rationally adjusted by the methods of agricultural pro-
duction and reinforced by the achievements of the scientific and
technical progress, first of all, by chemistry and microbiology
will remain the main source of nutrition for the population of
of our planet. This means that the main forces of science must
not be diverted from the agricultural production, but to concen-
trate on how to make it more profitable and productive, how to
utilize solar energy to the fullest, and how to increase the in-
dexes of food utilization of the total productivity of the bio-
sphere. We are deeply convinced that it is from these positions
that we should solve the problem where to use proteins of single-
cell organisms, in general, and hydrocarbon yeasts, in particular,
in the national economy in the near future. Moreover, we should
also remember that the population has a definite psychological
prejudice against various substitutes in the sphere of nutrition
and has an attachment to traditional foodstuffs to which they
are accustomed.
Thus, proteins of hydrocarbon yeasts must find application,
first of all, in the sphere of feeding agricultural animals. How-
ever, even in this case, protein-vitamin concentrates based on
these yeasts must satisfy sufficiently strict requirements which
would fully guarantee their safe use in the national economy.
We shall attempt to summarize the requirements for the
quality of hydrocarbon yeasts based on medico-biological studies.
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=mow' WOOF
Most of them can, e7-'idently be-achieved under the conditions of
a large-scale production. The list of preliminary requirements
for the quality of 3VK is given below.
Indexes of Chemical Studies
Content
Moisture Not over 10%
Total Nitrogen Not less than 8%
Content of
protein (N x 6.25) Not less than 48%
nucleic acid Not over 8%
It
lipids SI 5%
residual hyd-:rocarbons ? 0.1%
polycyclic hydrocarbons 0.000
Lead Not over 5 mg/kg
Arsenic II 5 mg/kg
Aflatoxins 0.000
Indexes of mic:robiological studies. Content
of microorganizos:
pathogenic 00
live yeast cells 00
nonpathogenic 100,000 per 1 g
Indexes of Bio1ogic.a-1 Studies
The most important requirement for the biological quali-
ties of nutrient yeasts is the absence of any kind of toxicity
when they are used, including acute or chronic toxicity, as well
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lapr,
wow
as the absence of cancerogenic, leukosogenic, teratogenic, and
mutagenic effects established on a number of biological species.
An even more necessary test of the safety of using hydro-
carbon yeast in feeding agricultural animals is the evaluation
of the quality of foodstuffs obtained from them. The most impor-
tant requirement in this case is the invariability of the chemi-
cal composition of foodstuffs and preservation of high organo-
leptic properties, and good technological and marketing conditions.
These products, when used regularly, must not have any ne-
gative effects on metabolism and, in particular, facilitate the
acceleration of the development of atherogenic processes in ag-
ing organisms.
The results of medico-biological studies described in this
book make it possible to assert that protein-vitamin concentrates
obtained in our country by utilizing the paraffin fraction of
petroleum even now satisfy, to a considerable degree, the above-
mentioned requirements. Also, it can be stated with assurance
that technological processes of the BVK production are being im-
proved constantly. One of the important goals of these improve-
ments is to improve the quality of BVK, to lower the content of
residual hydrocarbons in them, and to improve the stability of
the processes and conditions of yearly production, which, in the
final analysis, must lead to a higher guarantee of complete safe-
ty of their use in animal husbandry.
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Ng,
This conclusion is based on the detailed study of the dy-
namics of the accumulation of hydrocarbons with artificial intro-
duction of n-paraff ins in various quantities into BVK.
It was shown that, on the average, 1-2 percent of hydro-
carbons contained in re-'; tions can be retained in the fat tissue.
The processes of hydror_arbon accumulation progress slower during
the first half of the year and more intensively during the second
half of the year.
Thus, the fat t-:ssue may be defined as a kind of biologi-
cal trap accumulating hydrocarbons which get into the organism
from the environment.
We were able to confirm this later in our
studies at the agricul f-Airal animal fat depots.
Studies on the processes of hydrocarbon accumulation in
pigs revealed their inf:ignificant amounts in the subcutaneous
and internal fat only when the content of n-paraffins in BVK was
brought to 0.5 percent.
The above data f:nsure full prevention of the retention of
minimal amounts of hydrocarbon in the organisms of animals if the
recommendations of the Board of the USSR Ministry of Health are
followed in animal husidandrY.
Thus, the utili?Aation of hydrocarbon yeasts for the devel-
opment of animal farming may be the first great stride in the
development of the branch of microbiological industry producing
high protein products. Of course, it is also necessary to con-
sider the economic aspt, ct of the problem determining the expedi-
ency of producing nutrient hydrocarbon yeasts. However, the
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degree of economic expediency is determined by the concrete con-
ditions in each country, the availability of raw materials for
their production, costs of other sources of nutrient proteins,
and so on. Evidently, the profitability of their production is
beyond any doubt to the majority of European countries which have
to import fish and soy bean meal for animal food. At the same
time, in the U.S.A. the availability of considerable amounts of
inexpensive soy bean meal does not justify the production of pro-
tein-vitamin concentrates on the basis of hydrocarbon yeasts.
As a result of this, specialists of the U.S.A. are stressing the
development of studies on the use of microbiological proteins
directly in human diets.
We do not share this point of view and are not consider-
ing the introduction of proteins of unicellular organisms into
human diets in the near future. In our opinion, this concept is
connected with a considerable risk. However, scientific investi-
gations in this direction appear to be expedient. It should be
kept in mind that the biomass of unicellular organisms under cer-
tain conditions may become a necessary source for the production
of additional nutrient substances. For this reason, the reserves
of our country must also include methods for industrial process-
ing of unicellular organisms for foodstuffs.
The medico-biological investigations which have been car-
ried out, as well as the familiarization with the published data,
make it possible to formulate some preliminary requirements for
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Arne Nftwe
protein products produced from unicellular organisms which are
intended for food. It should be stressed that these considerations
are strictly preliminary. However, they, obviously, must be taken
into consideration to some degree by the scientists working in
this area.
In cases when it is intended to use proteins of unicellu-
lar organisms not for flavoring the food or additions to it, but
as a significant source of nutrient protein, these products can-
not be presented in the form of an unprocessed dry or moist bio-
mass of microorganisms, regardless of what structure or taste is
given to it. According to the available information, all attempts
to use washed biomass of various microorganisms in the human diet
usually produced definite complications connected with the pre-
sence in these preparations of high concentrations of nucleic
acids, unusual lipids, including sterols and ZhKN, biogenic
amines, unusual amino acids and peptides, amino sugars and muco-
polysaccharides, substances having a specific antigenic and al-
lergy-producing effect, and, finally, with the presence in some
of them of highly toxic substances specific for a given species
or even a strain of the microorganisms.
All this compels us to conclude that when attempts are
made to use the proteins of unicellular organisms as food, the
biomass of the microorganisms must be subjected to a rigid tech-
nological processing in order to isolate protein substances which
are purifiec: as much as possible. Some conditions for this
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Nor .ftor
process can be outlined even now. They should include, first of
all, the elimination of excessive amounts of nucleic acids. Evi-
dently, the maximum level of nucleic acid content in the isolated
protein substances must not exceed 1-2 percent; secondly, maximum
levels must be established for unusual biogenic amines, amino
acids of lipids and so on, for which it is still impossible to
establish any concrete values. It should be stressed that,evi-
dently, a considerable part of these substances can be removed
with the structures of cellular membranes which is technologically
possible and expedient. Thirdly, special attention must be given
to an almost total elimination of substances having a specific
amtigenic and allergy-producing effect; fourthly, close attention
nrzst be given to a total absence of the possibility of synthesis
by of toxic substances of the type of aflatoxins
ad other substances producing cancerogenic, teratogenic and mute-
genic effects; fifthly, sterility of the preparations, that is,
t:te absence of live producer cells in them; sixthly, a total ab-
sence of pathogenic microorganisms; seventhly, the establishment
norms for a possible contamination by nonpathogenic flora.
OtT course, the above-mentioned requirements for animal feed are
f-:zily applicable to the nutrient protein preparations.
Summing up the work done to date, we must stress that its
si.-.3-nificance is connected with the scientific and technical pre-
da.ztions regarding the ever increasing role of the microbiologi-
cz_i industry in the life of human society. There is no doubt
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Nei.)
that mankind is entering a new era of microorganism utilization.
Until recently, the attention to the microorganisms was attracted
because they were potential causative agents of diseases. Since
the moment of the discovery of antibiotics, microorganisms became
recognized as producers of powerful medicinal substances, and
this specialized branch of the microbiological industry has de-
veloped considerably.
At the present time, we are on the threshold of an exten-
sive introduction of microorganisms as a powerful productive force
into various branches of the national economy which may prove to
be extremely effective. One of the promising directions in the
production of unicellular organisms is their utilization as
unusually effective producers of protein substances. Evidently,
humanity must be sufficiently prepared for this possibility in
order to avoid significant errors and dangers connected with mak-
ing decisions insufficiently substantiated medically. The role
of medico-biological studies in this case increases greatly.
10,233
CSO: 8344/1098-W
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