Ergebnisse der Mikrobiologie, Immunitätsforschung und experimentellen Therapie: Fortsetzung der Ergebnisse der Hygiene, Bakteriologie, Immunitätsforschung und experimentellen Therapie

355 Pages·1957·14.4 MB·English

by  K. F. Meyer, E. G. Nauck, A. M. Pappenheimer Jr., J. Tomcsik (eds.)

ERGEBNISSE DER MIKROBIOLOGIE
IMMUNITATSFORSCHUNG
UND EXPERIMENTELLEN THERAPIE
FORTSETZUNG DER
ERGEBNISSE DER HYGIENE, BAKTERIOLOGIE,IMMUNITÄTSFORSCHUNG
UND EXPERIMENTELLEN THERAPIE
HERAUSGEGEBEN VON
w. KIKUTH K. F. MEYER E. G. NAUCK
OOSSELOORF SAN FRANCISCO HAMBURG
A.M. PAPPENHEIMER ]R. J. TOMCSIK
NEW YORK BASEL
DREISSIGSTER BAND
MIT 63 ABBILDUNGEN
SPRINGER-VERLAG BERLIN HEIDELBERG GMBH 1957
ISBN 978-3-662-23733-5 ISBN 978-3-662-25832-3 (eBook)
DOI 10.1007/978-3-662-25832-3
Alle Rechte, insbesondere das der t!bersetzung in fremde Sprachen, vorbehalten
Ohne ausdriickliche Genehmigung des Verlages ist es auch nicht gestattet, dieses
Buch oder Teile daraus auf photomechanischem Wege (Photokopie, Mikrokopie)
zu vervielfăltigen
© by Springer-Verlag Berlin Heidelberg 1957
Urspriinglich erschienen bei Springer-Verlag OHG. Berlin· Giittingen · Heidelberg 1957
Die Wiedergabe von Gebrauchsnamen, Handelsnamen, Warenbezeichnungen usw. in
diesem Werk berechtigt auch ohne besondere Kennzeichnung nicht zu der Annahme,
daB solche Namen im Sinn der Warenzeichen- und Markenschutz-Gesetzgebung
als frei zu betrachten wâren und daher von jedermann benutzt werden diirften
Inhaltsverzeichnis
Seite
I. BISSET, Dr. K. A., Birmingham. Multicellularity in Bacteria. With
11 figures and 2 plates . . . . . . . . . . . . . . 1
11. PIJPER, Professor Dr. ADRIANUS, Pretoria. Bacterial flagella and
motility. With 38 figures . . . . . . . . 37
III. BRANDIS, Privatdozent Dr. HENNING, Frankfurt a. M. Die Anwendung
von Phagen in der bakteriologischen Diagnostik mit besonderer Berück-
sichtigung der Typisierung von Typhus- und Paratyphus B-Bakterien
sowie Staphylokokken. Mit 10 Abbildungen . . . . . . . . . . . 96
IV. KAUFFMANN, Dr. F., Kopenhagen. Das Kauffmann-White-Schema 160
V. WELSCH, Professor Dr. MAURICE, Liege. Activites Bacteriolytiques
des Microorganismes . . . . . . . . . . . . . . 217
VI. RUSKA, Professor Dr. HELMUT, Albany (N. Y.). Die Bakteriencytologie
im Vergleich zum Feinbau höher differenzierter Zellen . . . . . . 280
VII. HENNEssEN, Dr. WALTER, Düsseldorf. Die serologische Diagnostik
der Viruserkrankungen des Menschen. Mit 2 Abbildungen. 288
Namen verzei chnis 317
Sachverzeichnis. 338
I. Multicellularity in Bacteria
By
K.A.BISSET
With 11 figures and 2 plates
Contents Page
1. Introduction . . . . . . . . . . 1
2. Organisation of the bacterial cell 7
3. The composition of the cell envelopes 13
4. Fission and cell division ..... . 1$
5. Techniques . . . . . . . . . . . . . 23
6. Multicellularity in relation to the systematics of bacteria 26
7. Summary 31
8. References . . . . . . . . . . • . . • . . . . . 33
1. Introduction
Advance in the study and understanding of the morphology and structure
of bacteria has been remarkably rapid in recent years. At the same time, it
has been, in a number of respects, exceedingly irregular. This irregularity has
manifested itself in diverse ways, although it certaillly cannot be held to betoken
any lack of general and widespread interest. For example, the degree to which
the more fully-accepted facts in the contemporary canon of knowledge on the
subject are understood is very different indeed in different cultural spheres.
Whereas, as might be anticipated, information of applied value, and especially
that of medical interest, is much more equably distributed.
A different type of irregularity in the distribution of information upon the
various aspects of bacterial cytology has an important bearing upon the subject
of this review. This is the rather remarkable discrepancy between the speed
at which information has spread and been assimilated, as between the different
aspects of investigation upon the problems of bacterial cytology, and between
the manner in which the information available upon different cellular com-
ponents has been accepted. The enormous preponderance of work has been
done upon the bacterial nucleus, and many, if by no means all the important
advances which have been made in this field are reasonably weIl known. And
although considerably less time and energy have been expended upon the flagella,
much of what has been discovered has been fully recognised by the majority of
those interested. The subject of what, for lack of a better term, may be called
'granular inclusions' in the bacterial cell, has also received a great deal of
attention, but, as will appear in the later part of this introductory section,
much of this work can onIy, howsoever reluctantly, be regarded as wasted; for
a reason, not uncommon in science, that the investigators in question have
sometimes failed to examine the premises of their arguments.
Ergebnisse der Mikrobiologie, Bd. 30 1
2 K. A. BISSET:
It may be that the attention which the foregoing problems have received
is due, at least in part, to the fact that all of them have been the subject of
controversies of a more or less violent nature. On the other hand, the one
major field in bacteriology which has not proved at all controversial in itself,
at least so far, has singularly failed to attract a similar degree of attention.
However, if controversy is indeed the best form of advertisement in science,
the problem of cellular structure in bacteria is not without hope of attaining
notoriety. Not because any real disagreement seems likely to a,rise among
those who have studied the matter, but because its proper understanding has
a most important bearing upon most, Ü not all, of those controversies concerning
the bacterial nucleus, flagella and granular inclusions, to which reference has
already been made. This will appear later, from reference to Text-fig. l.
It is a remarkable fact that although the existence of multicellular organi-
sation in bacteria has been known for the greater part of that period of a hundred
years which comprises the epoch of scientific bacteriology, it has never been
understood, nor indeed so much as recognised by the majority of bacteriologists.
It is now ten years since ROBINOW (81) published his remarkable testament of
bacterial cytology, and by his action advanced general knowledge both of the
nucleus and cellular structure of bacteria to a greater degree than has been
achieved by any single publication before or since. But despite the very large
number of papers dealing with problems in connection with the morphology,
complexity, growth and composition of the cell envelopes of bacteria which has
appeared since that date, many bacteriologiflts, even including a surprisingly
high proportion of those actually engaged in the study of cytology, persist in
treating bacteria as single cells, and in referring to them as such.
This does not apply only to those bacteria that, from lack of positive in-
formation to the contrary, may be regarded as single cells, either on the principle
of choosing the simpler of two alternatives, or by analogy with known, uni-
cellular genera, to which the bacteria under discussion may bear some real or
fancied resemblance. It is also true of those genera, including most, if not all
Gram-positives, the evidence for whose multicellular nature is beyond all dispute.
No better example than this could be found of the well-known principle that
most observers, scientüic or otherwise, see only what they expect to see. Or
more explicitly, what they have been taught to expect to see. For if once
familiarity with the concept has been gained, the division of, for example, a
staphylococcus into two or four cells can easily be discerned in a Gram-stained
preparation with a scholastic oil-immersion lens. Despite which, generations of
bacteriologists have trained their successors toregardit as an undifferentiated sphere.
The amount of work which has been done upon the basis of this fallacious
premise ha.s automatically given rise to what can only be termed avested interest
in the traditional, unicellular concept. This is the sort of situation which occurs
in all branches of science, and will doubtless continue to do so. In the present
case, it is obvious that where the supporters of a particular school of thought
have staked their professional reputations upon some interpretation founded
upon the dogma that a bacterium is a single rod-shaped cell, they will tend
to regard without enthusiasm the suggestion that many bacteria are, in fact,
filaments of smaller cells, separated by cross-walls.
Multicellularity in Bacteria 3
This is peculiarly true of those workers who rely too exclusively upon the
electron microscope. This apparatus has provided some valuable information
concerning the surface structures of bacteria, and especially concerning the
flagella, but it is liable to provide a misleadingly convincing picture of uni-
cellularity in bacteria which can clearly be seen, by the use of suitable cyto-
logical techniques with the classical light microscope, to be markedly multi-
cellular. It must be admitted that, upon occasion, internal cell boundaries can be
discerned in disrupted cells by this means [ANGELICO etal. (1), DAWSON and STERN
(38), YOSHIDA, FUKUYA et al. (106)] but even in such cases it is a matter for
question whether more than a small proportion of these structures can actually
be rendered visible. Comparative studies by light and electron microscopy
suggest that they cannot [BISSET (23)].
Too exclusive a reliance upon any single technique, not only upon the electron
microscope, can lead to errors and misconceptions, especially if this narrow
approach is to some degree guided by a preconceived view of the objective.
It is axiomatic that a scientific hypothesis should be designed to explain observed
facts, and that subsequent research should be designed to test the hypothesis.
But it would be idle to pretend that these rules are invariably followed. It has
happened repeatedly in bacterial cytology that workers employing a single tech-
nique, or a group of techniques directed to a single purpose, have promulgated
some rash interpretation, usually by analogy with other types of cell, and have
then expended much subsequent time and eHort in the design of experiments
intended, not so much to test their hypotheses as to protect them against incon-
venient facts adduced from other sources by other workers. An excellent example
of the spirited defence of an elegant theory against intrusive and incompatible
evidence is given by HUNTER (56). This example is of peculiar interest in the
present context, since, as in so many parallel cases, the original flaw in the
theory of nuclear structure proposed by this worker and her collaborators was
that the Gram-positive coccus under examination proved not to be a single cell,
as was essential to the validity of the theory, but a complex of several cells,
divided by septa [BISSET (19)]. Although this muIticellularity in cocci has since
been verified by a variety of methods [WEBB and CLARK (101), TOMCSIK and GRAC
(92)], HUNTER has attempted to obviate this criticism of her work by demon-
strating that it is possible so to stain the coccus in question that its cross-walls
are not revealed. It is fair to comment that by exactly comparable means it
has been proved repeatedly that bacteria are devoid of an organised nucleus!
FaiIing to demonstrate the cytological structures of the bacterial cell presents
no difficulties; the problem is to avoid such failure.
Even in the limited field of the cocci a number of further examples can be
found of theories of nuclear behaviour which are destined to make shipwreck
on the selfsame rock. One of these is that of KRIEG (61), who manifestly mistakes
entire cells, stained with fluorescent dyes, for nuclei in a septate coccus which
he assumes, in the absence of any evidence whatsoever, to be unicellular. Once
more, too exclusive a reliance upon a single technique has proved dangerous. The
:r:arallel application of a cell wall stain, a relatively simple matter in the
case of most cocci, would have supported or disproved KRIEG'S ideas (Plate I,
Fig.8).
1*
4 K. A. BISSET:
The examples just quoted illustrate how, in the absence of an understanding
of the multicellular character which is typical of the majority of Gram-positive
and some Gram-negative bacteria, errors may arise from failure to realise that
all the components visible in a single coccus or bacillus may not be, indeed
almost certainly are not, parts of a single ceH, but usually belong to several
different cells, and may comprise the major portions of small cells. This mis-
conception has led especially to errors in interpretation of the nuclear behaviour
of bacteria, but turns up also in the 'diagnostic' staining of bacteria [cf. BISSET (11)].
A second, parallel error arising from the same source, consists in the mistaking
of elements which are actually derived from the cell envelopes for internal
structures lying within the cello This occurs in studies both of cocci and bacilli,
but especially the latter. The septa and cross-waUs which subdivide the multi.
cellular bacteria may possess, or be associated with elements containing con-
siderable components of nucleic acids. These are capable of combining with a
variety of different types of basic dye, and frequently the most obvious structures
demonstrable in this fashion are the granules at the junction of the cell wall
or membrane and the developing cross-walls. These are clearly shown in the
illustrations of many reliable cytologists, as for example KNAYSI (60), JÄRVI and
LEVANTO (58), and ROBINOW and MURRAY (82). The simple and straightforward
interpretations of these authors, with which the present writer entirely concurs,
are shown in Text-fig. 1 (a, b, cl.
Also in Text-fig. 1 are shown some of the numerous alternative inter-
pretations which have been made, by different authorities, to explain what are,
beyond any reasonable doubt, the same structures. And it is quite obvious,
not only that many of these are incompatible with the interpretation which has
already been mentioned, and which is almost certainly the true one, but that they
are also incompatible one with another. Indeed, in some cases, as will be ob ..
served, two or more interpretations proposed by the same author or authors
may be in themselves mutually contradictory. The internal contradictions appear
mainly to be due to a type of uncritical reliance upon a single technical method,
supposed, often enough manifestly erroneously, to be specific for some particular
element of the cello Portions of the bacterial cell envelopes are capable of being
stained by techniques stated by various authorities to be reliable methods of
distinguishing, for example, nuclei, mitochondria, fat globules, and even reticulo-
cytes [BISSET (17)].
The first three interpretations shown in Text-fig. 1, those of KNAYSI,
JÄRVI and LEVANTO, and ROBINOW and MURRAY, have already been referred to.
That of DELAMATER and MUDD (40) is shown only diagrammaticaHy here (d). In
the various figures published by these authors, the 'centrioles' which they claimed
to demonstrate lie always in one or another of the sites illustrated in the figure;
either on the mid-line at the poles of the bacillus, or in the comparable position
on the complete cross-wall which represents the forthcoming fission of the
bacillus in the centre; or more commonly at the junction of the cell wall and the
cross-walls.
Stained by a different method, the material surrounding the cross-wall was
equated with mitochondria (e) by MUDD and WINTERSHEID (73). On the other
hand McGREGOR (67), while agreeing with the previous workers in their inter-
Multicellularity in Bacteria 5
(a) KNAYSI (1930) (: 11~ : )
(b) JÄRVI and LEVANTO (1949) Development of cell envelopes
GD
(c) ROBINOW and MURRAY (1952) (=:]11 : ~
(d) D
(1E9L5A1M) ATER and MUDD €: : : ) Centrioles
(e)
(M1U95D3D) and WINTERSHEID ( 1I1 ) Mitochondria
(f) MOGREGOR (1954) r.... ~Nuclei
~ • 7--- Centrioles
--------
(g) CHAPMAN and HILLIER (1953) 0 '---...J\._ ___ Developing septa
(h) MUDD (1953) .f------\-b Mitochondria
(i) DAVIS and MUDD (1953) • I~NuclMeitioch ondria
Foci for:
(j) WEIBULL (1953) •
~ • ~
Redox reagent.,
=-
(k) BISSET (1953)
I Intrinsic and
Extrinsie DNA
(I) BISSET (1954)
~ ~ ~evel~Ping septa
(m) DELAMATER (1954) ~ _ CentrlOles
Mitoehondria
(0) f)~~; ""' G",,-H"~" (:::JlCY Development of eapsular septa
Areas of acti ve growth. Serve
as foei, and stain with a wide
(0) Summary of evidenee, to date -.·:.:-:-1.·. . :-1::::.1::.:_ range of supposedly speeifie eyto-
ehemieal reagen ts
Text-fig. 1. Interpretations from baeterial cell envelopes. The figure shows diagrammatieally the basophilic
areas and granules which are commonly found at the points of cell division in septate baeteria, and the very
diverse interpretations whieh have been based upon these appearanees. Some of these, for example a, b, c, g,
i, k, I, n, are sound and mutually corroborative. Others are, for the most part, fanciful and mutually exelusive.
Diseussion in text
pretation of the granules at the poles of the cell as centrioles, chose to regard
those surrounding the cross-wall as nuclei rather than as mitochondria or further
centrioles (f).
Ergebnisse der Mikrobiologie, Bd. 30 la
6 K. A. BISSET:
A great deal of valuable information concerning the bacterial cell envelopes
has been obtained from the remarkable and beautiful electron micrographs of
ultra-thin sections produced by CHAPMAN and HILLIER (35). These authors inter-
preted their results (g) in terms very closely comparable with those of KNAYSI,
JÄRVI and LEVANTO, and ROBINowand MURRAY, with certain exceptions which
will be discussed in a later section. But MUDD (71), describing exactly the same
material, (h), on ce more chose to interpret as mitochondria the elements of
developing cross-walls which CHAPMAN and HILLIER, in accordance with the con-
sensus of reliable opinion on the subject, had clearly described as cell-envelope
material. DAVIS and MUDD (37) reserved the title of mitochondria for the sm aller
elements at the poles of the bacillus, and at the junction of cell wall and cross-
walls, (i). The larger masses, disposed around the central cross-wall, they de-
scribed as nuclei. These protagonists of bacterial mitochondria were attacked
by WEIBULL (102), who showed that the supposedly specific reagents, which
they used for their demonstrations, formed granular aggregations in exactly
the same sites, (j). And this observation was paralleled by those of BISSET (17,
18) who showed firstly that stainable materials with the reactions of DNA tended
to become transferred from the nucleus, and possibly also from the cytoplasm
of bacteria, under the influence of cytological techniques, and to form aggregates
at the poles and cross-walls, (k). Secondly, he showed that DNA of extrinsic
origin, derived in this case from herring-roe, when brought into physical contact
with similar bacteria, appeared in the form of clusters of small granules at
exactly the same places, and that some of these artifacts very closely simulated
the types of controversial cytological elements with which this section is con-
cerned (l). More recently DELAMATER (39) simultaneously described different
portions of the polar and cross-wall elements as mitochondria and centrioles,
in accordance with his previous views, and also as developing septa, in accordance
with the criticisms of those of his opponents who had pointed out that this was,
Itlmost certainly, the true nature of his 'mitochondria' and 'centrioles', previously
described, (m). This confusion is commented upon by MARSHAK (66), who points
out the dubiety of claims to draw important distinctions on the ground that
these tiny granules stain in different shades of purple. A further vindication of the
views illustrated in a, band c was provided by TOMCSIK and GUEx-HoLZER (96),
who showed that those areas which were regarded by the authorities previously
quoted and by BISSET (17, 21) as the areas of growth of the cell envelopes, are
certainly the areas in which the main growth of the capsule occurs, (n). And since
the conclusions of TOMCSIK and GUEx-HoLZER were derived, not from stained and
fixed preparations, with all the limitations which these conditions imply (as
can be seen quite clearly from the discrepant views recorded in the diagram),
but from the examination by phase-contrast microscopy of living bacteria treated
with antibodies, the support which they provide is peculiarly valuable.
The final diagram (0) summarises what is really known about these areas
in the developing, multicellular bacillus, and enables a distinction to be drawn
between reasonable theories, supported by mutually corroborative evidence, and
pure speculation, often mutually contradictory in a high degree, and supported
only by analogy. These basophilic 'granules' are believed to correspond with the
divisions between the cells of a multicellular rod, and mark the areas of growth